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High consumption of ultra-processed food (UPF) has been associated with increased risk of obesity and other metabolic diseases, and this dietary pattern seems to be responsible for chronic changes in the gut microbiota. The aim of this study was to assess the associations of UPF with the gut microbiota and obesity-associated biometrics in women.This cross-sectional study examined 59 women. The following parameters were evaluated: food consumption using NOVA classification, anthropometric and metabolic parameters, and gut microbiome by next-generation sequencing. The mean age was 28.0 ± 6.6 years. The mean caloric intake was 1624 ± 531 kcal, of which unprocessed or minimally processed food (G1) accounted for 52.4 ± 13.5%, and UPF accounted for 31.4 ± 13.6%. Leptin levels adjusted for fat mass were negatively associated with G1 and positively associated with UPF. We found 15 species in the gut microbiota that correlated with G1 (3 positively and 12 negatively) and 9 species associated with UPF (5 positively and 4 negatively).Higher consumption of UPF was directly associated with leptin resistance, and this study suggests that the consumption of UPF or G1 may affect the composition of the gut microbiota.

High hyperdiploid acute lymphoblastic leukemia (HeH ALL), one of the most common childhood malignancies, is driven by nonrandom aneuploidy (abnormal chromosome numbers) mainly comprising chromosomal gains. In this study, we investigate how aneuploidy in HeH ALL arises. Single cell whole genome sequencing of 2847 cells from nine primary cases and one normal bone marrow reveals that HeH ALL generally display low chromosomal heterogeneity, indicating that they are not characterized by chromosomal instability and showing that aneuploidy-driven malignancies are not necessarily chromosomally heterogeneous. Furthermore, most chromosomal gains are present in all leukemic cells, suggesting that they arose early during leukemogenesis. Copy number data from 577 primary cases reveals selective pressures that were used for in silico modeling of aneuploidy development. This shows that the aneuploidy in HeH ALL likely arises by an initial tripolar mitosis in a diploid cell followed by clonal evolution, in line with a punctuated evolution model.

High hyperdiploid acute lymphoblastic leukemia (ALL) is one of the most common malignancies in children. The main driver event of this disease is a nonrandom aneuploidy consisting of gains of whole chromosomes but without overt evidence of chromosomal instability (CIN). Here, we investigated the frequency and severity of defective sister chromatid cohesion—a phenomenon related to CIN—in primary pediatric ALL. We found that a large proportion (86%) of hyperdiploid cases displayed aberrant cohesion, frequently severe, to compare with 49% of ETV6/RUNX1‐positive ALL, which mostly displayed mild defects. In hyperdiploid ALL, cohesion defects were associated with increased chromosomal copy number heterogeneity, which could indicate increased CIN. Furthermore, cohesion defects correlated with RAD21 and NCAPG mRNA expression, suggesting a link to reduced cohesin and condensin levels in hyperdiploid ALL. Knockdown of RAD21 in an ALL cell line led to sister chromatid cohesion defects, aberrant mitoses, and increased heterogeneity in chromosomal copy numbers, similar to what was seen in primary hyperdiploid ALL. In summary, our study shows that aberrant sister chromatid cohesion is frequent but heterogeneous in pediatric high hyperdiploid ALL, ranging from mild to very severe defects, and possibly due to low cohesin or condensin levels. Cases with high levels of aberrant chromosome cohesion displayed increased chromosomal copy number heterogeneity, possibly indicative of increased CIN. These abnormalities may play a role in the clonal evolution of hyperdiploid pediatric ALL.

This study aimed to characterize the effects of dietary restriction and subsequent re-alimentation on body composition and hepatic gene expression of epigenetic markers of DNA methylation, RNA m6A methylation, and histone acetylation in the liver of postpubertal beef bulls. Twelve Angus × Hereford crossbred bulls (n = 6, 23 ± 0.55 mo [young bulls], 558 ± 6.1 kg; and n = 6, 47 ± 1.2 mo [mature bulls], 740 ± 30.5 kg) were submitted to two dietary regimes per offering of the same hay: low plane of nutrition (90 d) and compensatory growth (90 d). Each animal acted as its own control and were fed Beardless wheat (Triticum aestivum) hay and mineral mix during the trial. Statistical analyses were performed using SAS 9.4 following a pre-post repeated measures design. Bulls in negative energy balance (NEB) decreased (P0.001) empty body weight (EBW; 23.1% [-139.1 kg]), empty body fat (EBF; 39.8% [-85.4 kg]), and empty body protein (EBP; 14.9% [-13.5 kg]) and fully recovered at the end of the trial. Body fat accounted for 77.1% of daily changes in body energy status, whereas body protein accounted for only 22.9% (P0.001). Relative abundance of epigenetic markers transcripts was analyzed via qPCR. Bulls at NEB tended (P ≤ 0.097) to increase gene expression of epigenetic markers of RNA m6A methylation (METTL14, VIRMA, and WTAP) and increased (P ≤ 0.050) the gene expression of epigenetic markers of DNA methylation (DNMT3A) and histone-acetylation (SIRT3 and SIRT7). Young bulls had a tendency (P ≤ 0.072) of higher RNA m6A methylation, VIRMA, and WTAP than mature bulls. Effect of diet × age interaction was not detected (P ≥ 0.137) for METTL14, VIRMA, WTAP, DNMT3A, SIRT3, or SIRT7. Younger bulls tended to have greater RNA m6A methylation levels than mature bulls, indicating that, while contemporaneously fed the same diet during periods of undernourishment followed by compensatory growth, age has an impact on this epigenetic mechanism. In conclusion, metabolic status seems to carry a greater impact on regulating bovine hepatic epigenetic mechanisms that modulate gene transcription, such as DNA methylation and histone acetylation, than on epigenetic mechanisms that regulate gene translation, such as RNA m6A methylation. During periods of undernourishment followed by compensatory growth, body fat pools appear to change more dynamically and are easily detected having a greater impact on epigenetic markers that modulate hepatic gene transcription rather than translation.Epigenetics refers to heritable modifications that regulate gene expression without altering DNA sequence, hence, acting on top of the genes. Epigenetic markers change in response to stressors such as environmental factors, nutritional challenges, among other overlooked players that altogether could drastically impair animal performance. During periods of undernourishment followed by fast weight gain, dynamic changes in body composition, especially fat, appear to trigger an increased action of such physiological markers that modulate hepatic gene expression. Findings of this study unveil epigenetic metabolic pathways that deserve further investigation for proper quantification of potential consequences of metabolic stress on the liver of bovines that suffer significant loss of body weight followed by recovery. The alterations at the molecular level shown in this study provide a picture of silent metabolic changes that have not been detected previously in liver metabolism studies of cattle. Therefore, the impact of nutritional management and metabolic stress may be greater than previously expected and differently controlled than previously assumed.

Aiming to characterize the effects of nutritional status on epigenetic markers, such as DNA 5-methyl cytosine (mC) methylation and RNA N6-methyladenosine (m6A) methylation, of bovine sperm, 12 Angus × Hereford crossbred breeding bulls were submitted to nutritional changes for a period of 180 d: no change in body weight (BW) (phase 1 = 12 d), BW loss (phase 2 = 78 d), and BW gain (phase 3 = 90 d) in a repeated measures design. Animals were fed Beardless wheat (Triticum aestivum) hay and mineral mix. Statistical analyses were performed using SAS 9.4 (SAS Inst., Cary, NC). Higher levels of RNA m6A (P = 0.004) and DNA methylation (P = 0.007) of spermatic cells were observed at phase 2 compared with phase 1. In phase 3, sperm RNA m6A methylation levels continued to be higher (P = 0.004), whereas the DNA of sperm cells was similar (P = 0.426) compared with phase 1. Growing bulls had a tendency (P = 0.109) of higher RNA m6A methylation levels than mature bulls. Phase 2 altered scrotal circumference (P < 0.001), sperm volume (P = 0.007), sperm total motility (P = 0.004), sperm progressive motility (P = 0.004), total sperm count (P = 0.049), normal sperm (P < 0.001), abnormal sperm (P < 0.001), primary sperm defects (P = 0.039), and secondary sperm defects (P < 0.001). In phase 3, bulls had scrotal circumference, sperm volume, sperm motility, sperm progressive motility, total sperm count, normal and abnormal spermatozoa, and primary and secondary spermatozoa defects similar to phase 1 (P > 0.05). Serum concentrations of insulin-like growth factor-1 and leptin decreased during phase 2 (P = 0.010), while no differences (P > 0.05) were detected between phases 3 and 1; growing bulls tended (P = 0.102) to present higher leptin levels than mature bulls. Specific for mature bulls, DNA methylation was positively correlated with leptin concentration (0.569, P = 0.021), whereas for young bulls, DNA methylation was positively correlated with abnormal spermatozoa (0.824, P = 0.006), primary spermatozoa defect (0.711, P = 0.032), and secondary spermatozoa defect (0.661, P = 0.052) and negatively correlated with normal spermatozoa (−0.824, P = 0.006), total sperm count (−0.702, P = 0.035), and sperm concentration (−0.846, P = 0.004). There was no significant correlation (P > 0.05) between RNA m6A and hormones and semen traits. In conclusion, the nutritional status of breeding bulls alters epigenetic markers, such as DNA methylation and RNA m6A methylation, in sperm, and the impact of change seems to be age dependent. These markers may serve as biomarkers of sperm quality and fertility of bulls in the future. Detrimental effects on sperm production and seminal quality are observed at periods and places when and where environmental and nutritional limitations are a year-round reality and may carry hidden players that may influence a lifetime of underperformance.

Hyperdiploidy, i.e. gain of whole chromosomes, is one of the most common genetic features of childhood acute lymphoblastic leukemia (ALL), but its pathogenetic impact is poorly understood. Here, we report a proteogenomic analysis on matched datasets from genomic profiling, RNA-sequencing, and mass spectrometry-based analysis of >8,000 genes and proteins as well as Hi-C of primary patient samples from hyperdiploid and ETV6/RUNX1-positive pediatric ALL. We show that CTCF and cohesin, which are master regulators of chromatin architecture, display low expression in hyperdiploid ALL. In line with this, a general genome-wide dysregulation of gene expression in relation to topologically associating domain (TAD) borders were seen in the hyperdiploid group. Furthermore, Hi-C of a limited number of hyperdiploid childhood ALL cases revealed that 2/4 cases displayed a clear loss of TAD boundary strength and 3/4 showed reduced insulation at TAD borders, with putative leukemogenic effects., High hyperploidy is a common feature in childhood B-cell precursor acute lymphoblastic leukemia. Here, the authors perform proteogenomic and Hi-C analyses of this leukemia and the ETV6/RUNX1 subtype and show that CTCF and cohesin expression are low in hyperdiploid cases and transcriptional dysregulation in relation to topologically associating domain borders in some of these cases.

Um processo judicial é formado, em essência, por uma sequência de atos que são traduzidos em movimentações processuais que carregam o histórico do desenvolvimento das atividades realizadas pelas mais diversas partes no curso da demanda processual. Neste trabalho, explicitamos os caminhos mais prováveis que caracterizam as classes de processos judiciais por meio de um conjunto de regras atribuídas a uma rede complexa. Observamos a sequência de atividades mais prováveis e destacamos os tempos médios necessários para serem concluídas. Observamos quais seriam os principais pontos de atrasos nas possíveis formas de inicialização e término de um processo além da similaridade entre os possíveis caminhos tomados. Ademais, indicamos qual seria o tempo gasto, em média, no aumento do número de movimentações para cada classe de processo. A caracterização dessas redes e a contextualização do caminho de movimentação ideal, associado aos outros dados gerados pelo método proposto permitem, por meio de um processo confiável de geração de dados, uma observação detalhada do fluxo dos processos judiciais, a identificação de comportamentos atípicos e o planejamento de intervenções capazes de otimizar a prestação do serviço jurisdicional pelo judiciário.

Monoethylene glycol (MEG) is a gas hydrate inhibitor widely applied for natural gas flow assurance. A series of density and electrical conductivity measurements of water + MEG + NaCl mixtures are reported, allowing the supervision of the MEG regeneration unit. Density (509 data points) and electrical conductivity (212 data points) measurements were performed in wide ranges of temperature, T = 278.15−363.15 K, and concentration of solvents and NaCl up to almost saturation. The theory of solutions was applied for density description using excess volume, which was correlated with the Redlich−Kister equation. The resulting absolute and relative mean deviations are 0.00127 g·cm−3 and 0.12%, indicating accurate representation. A semi- empirical correlation with 15 adjustable parameters was considered for electrical conductivity of water + MEG + NaCl mixtures. The obtained absolute and relative mean deviations are 1.49 mS·cm−1 and 5.70%. The properties functions presented an approximately orthogonal behavior to each other, allowing the determination of mixture composition from experimental density and electrical conductivity data. The Matlab environment was found to be robust in solving the nonlinear system of two equations with constraints. The proposed methodology was extensively tested, and deviations less than 0.0060 and 0.0011 in solvents and NaCl mass fractions were obtained, respectively, demonstrating the required accuracy for industrial application

2030-12 Monoethylene glycol (MEG) is used to prevent hydrate formation in subsea pipelines of natural gas production. MEG is recovered at an offshore regeneration unit. Sodium chloride is the predominant salt in the water−MEG streams. Thus, knowledge of NaCl solubilities in the process conditions is relevant for the design and operation of a MEG regeneration unit. Six isotherms of NaCl solubility were measured (76 data points) from T = (293.15 to 403.15) K in the whole solvent concentration range. The analytical method for salt solubility was based on the reproducibility of the concentration of two successive samples. Density measurements were accurately applied for these analyses. The temperature dependence on NaCl solubility is relatively weak. A reverse behavior with increasing temperature was observed for MEG contents higher than 90 wt %. Salt solubilities were successfully correlated with Redlich−Kister expansion (AADwNaCl = 0.0006 and ΔwNaCl = 0.47%). Electrical conductivity of NaCl - saturated solutions were measured (44 data points) at T = (293.15 to 363.15) K and correlated with a semiempirical model. A large difference in electrical conductivity for the two solvents was observed. The correlation adequately represented the data set (AADκ = 5.67 mS·cm−1 and Δκ = 5.94%). NaCl solubilities and electrical conductivities were accurately determined and are useful for the MEG regeneration process

The study of coals affected by magmatic intrusions is important from a geological point of view, in order to understand how heat and magmatic fluids could affect the organic and inorganic fractions as well as the properties of the coals. These fluids may lead to an enrichment of trace elements and heavy metals of environmental concern or serve as unconventional sources of critical elements. Therefore, the objective of this study is to investigate the organic and inorganic fractions of Peñarroya-Belmez-Espiel Basin (PBEB) (Spain) coals in order to verify whether changes of the petrography, chemical composition, mineralogy, and inorganic geochemistry have taken place. The PBEB corresponds to a Carboniferous intramontane basin filled with a Middle Mississippian to Lower-Middle Pennsylvanian sedimentary succession with coal-bearing units. Two main successive depocentres (extended from NW to SE) were recognized and in which bituminous and anthracite coals, as well as natural cokes occur. Petrographic analysis, ultimate and proximate analysis, scanning electron microscopy with energy dispersive X-ray spectrometry (SEM-EDS) analysis, inductively coupled plasma-optical emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS) analysis were carried out for the purpose of this study. The sampling consisted of a selected set of nine samples of bituminous coals, ten samples of anthracites and three samples of natural cokes. The results show that the organic fraction of bituminous and anthracite coals is mainly made up of vitrinite. Although the minerals identified in the two depocentres vary, they mainly consist of quartz and clay minerals in both cases. Authigenic kaolinite and epigenetic mineralization related to thermal events that occurred in the basin were observed in the anthracites. Natural coke shows isotropic and fine mosaic textures, and signs of epigenetic mineralization. The trace element content of the coal samples was compared with the values established for world hard coals and an enrichment of Cs, in bituminous and anthracite coals, and of Hg in natural cokes, was observed. The occurrence of REY in bituminous coals is related to the detrital sedimentary source, whereas in anthracite the epigenetic and thermal effects may have also been involved. In natural cokes, the REY content is probably associated with the original coal. Mercury enrichment in natural cokes may be associated with magmatic fluids and explained by Hg volatilization, its capture in natural coke pores and subsequent precipitation., The author H. Moura benefited from a doctoral scholarship financed by the FCT - Fundação para a Ciência e a Tecnologia (Ref: SFRH/BD/134109/2017). This work was financed by national funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., under the project Ref. UIDB/04683/2020.

Agriculture is by far the largest consumer of water, with about 70% of the diverted water being used in irrigation. Agriculture is also considered as a key source of diffuse pollution with inefficient practices resulting in high water and nutrient (particularly N and P) surpluses that are transferred to water bodies through diffuse processes (runoff and leaching), promoting eutrophication with associated biodiversity loss. WATER4EVER aims to demonstrate the connectivity between agricultural practices carried out at the plot scale and the resulting environmental effects that are usually first perceived further downstream. For that, the project combines the most performant monitoring strategies at the plot scale to provide detailed information of water flow and nutrient transport, integrating then this information at the catchment scale to close the gap between diffuse loads and water quality degradation. The specific objectives are: (i) to develop an automatic irrigation and fertilization Decision Support System (DSS) based on online data and forecast models; (ii) to develop new strategies for continuous monitoring of crop development based on optical sensors installed on fixed and mobile ground, drone and satellite platforms; (iii) to improve process-based models for the dynamics of water and its implication on nutrient budgets, crop development, and soil erosion; (iv) to connect plot and catchment scale models to quantify the effect of local agriculture practices on downstream water availability and quality; and (v) to disseminate and transfer knowledge and technology by means of public information adapted to different stakeholders. WATER4EVER was implemented in multiple case studies in Portugal, Spain, Italy, and Turkey. Earlier work was on the collection of field datasets for the calibration/validation of some of the innovative tools and approaches developed in the project. These included: (i) a modular smart camera for crop monitoring, which was integrated with other modular and open-source IoT based technologies to upgrade conventional machinery with variable rate technologies to reach higher levels of precision for crop monitoring; (ii) irrigation scheduling tools for improving irrigation water use efficiency in water scarce regions; (iii) the IrrigaSys DSS for irrigation water management at the plot scale based on online, open source tools; (iv) 1 km soil moisture maps for monitoring surface soil moisture at the catchment scale; (v) 100 m soil moisture maps for plot scale monitoring of soil moisture conditions; and (vi) vegetation products (LAI and NDVI) (30m resolution) for crop growth monitoring. Project results further included: (i) validated protocols for sustainable irrigation water management; (ii) guidelines on the best practices for integrating earth observation data in model-based irrigation and fertilization optimization strategies; and (iii) guidelines on the best practices related to soil and water management for each case study. WATER4EVER results can contribute directly to the implementation of the Nitrates Directive and the Water Framework Directive. The Project is particularly dedicated to raising awareness on the causes of diffuse pollution and respective impacts on the availability and quality of downstream water bodies; on reducing costs of production factors by improving irrigation water and fertilizers efficiency; and on demonstrating the capabilities of emerging technologies (smart cameras, satellite data, process-based models) for improving irrigation water and fertilization management. WATER4EVER is led by Instituto Superior Técnico (Portugal) and includes Deimos Engenharia SA (Portugal), Instituto de Engenharia de Sistemas e Computadores, Tecnologia e Ciência (Portugal), isardSAT (Spain), Universidad Politécnica de Cartagena (Spain), Institute for Agricultural and Earthmoving Machines of the National Research Council of Italy (Italy), and the Abant Izzet Baysal University (Turkey) in the consortium. More information on WATER4EVER is available at http://water4ever.eu/.

2030-12-30 Vapor−liquid equilibrium (VLE) data for aqueous systems in the presence of electrolytes have many industrial applications. VLE data for water + monoethylene glycol (MEG) mixtures in the presence of sodium chloride at low pressures are important to describe the effects of composition, temperature, and pressure on a MEG (gas hydrate inhibitor) regeneration unit. A modified version of the Othmer ebulliometer was applied to measure reliable VLE data for water + MEG + NaCl at 101.325, 65, and 35 kPa. Binary systems (water + MEG and MEG + NaCl) were also experimentally studied. Vapor pressures for water and MEG were determined and compared with the selected literature data via Antoine correlation. The electrolyte nonrandom two-liquid and universal quasi-chemical activity coefficient models were successfully parameterized to describe the VLE behavior for water + MEG + NaCl systems. Thermodynamic consistence of the data sets was also checked. Interaction parameter estimation followed a systematic strategy: (1) water−MEG, (2) water−NaCl, and finally (3) MEG−NaCl with the experimental data of MEG + NaCl and ternary data. MEG + NaCl solutions presented an inverted colligative property, that is, the addition of salt decreases the boiling point. VLE data indicated that water separation is less efficient due to the addition of salt. The parameterized models allow an evaluation of the MEG regeneration process as a function of temperature, pressure, and composition

Sistemas de informações em saúde (SIS) vêm aumentando em complexidade e volume de dados. Os dados nem sempre seguem um padrão, agravando a integração de serviços, especialmente quando envolve sistemas legados. Este trabalho propõe a PIXEL, uma plataforma para integrar experimentos de interoperabilidade em sistemas legados de saúde pública. Essa solução foi aplicada ao GISSA, um sistema inteligente na área da saúde que utiliza aprendizagem de máquina para gerar alertas e apoiar à tomada de decisão de seus usuários. A PIXEL realizou a integração de sistemas e implementa os três níveis iniciais de interoperabilidade, conforme proposto pela Society for Health Information and Management Systems (HIMSS).

High hyperdiploid (HeH) B-cell precursor acute lymphoblastic leukemia (BCP ALL) is characterized by a very specific nonrandom gain of chromosomes; a feature distinct from other types of aneuploid tumor types that usually display very heterogeneous gains and losses of chromosomes. Gains of chromosomes X, 4, 6, 10, 14, 17, and 18 are seen in more than 75% of cases of HeH childhood ALL, and of chromosome 21 in 100% of cases. In contrast to many aneuploid malignancies, there has been little evidence of chromosomal instability (CIN) in HeH ALL and the mechanisms leading to these chromosomal gains remain unknown. The aim of this project was to determine the level of genomic heterogeneity in HeH ALL. In order to do this, we performed low-pass whole genome sequencing (WGS) of single cells isolated from diagnostic bone marrow samples from HeH ALL patients to investigate cell-to-cell heterogeneity. Single nuclei in G 1 phase from nine diagnostic childhood HeH ALL samples were isolated using fluorescence-activated cell sorting and DNA libraries were constructed for low-pass WGS. Copy number analysis for each individual cell was performed in-house using the software programs AneuFinder and Ginkgo. Homolog inheritance of chromosomes gained or lost was determined by screening for heterozygous variants and calculation of the variant allele frequencies (VAF). Sequencing 2,572 single cells showed that the nine HeH ALL patients were all relatively homogenous at the trunk of their evolution tree and that the bulk of chromosomal gains were stable and unchanging. Structural aberrations, visible as partial chromosome copy number changes, were detected at diagnosis in most cases, with a greater number of structural aberrations detected in patients with greater numbers of sub-clones (defined as three or more cells presenting with the same numerical and structural aberrations). Unique numerical chromosomal aberrations detected in two or fewer cells were relatively random and therefore indicative of nondisjunction events in recent cell divisions rather than being part of the clonal evolution of the leukemia. These results indicate very low heterogeneity in HeH ALL and suggests that the genome of these leukemias is relatively stable. Disclosures No relevant conflicts of interest to declare.

Monoethylene glycol (MEG) is used as hydrate inhibitor in natural gas exploitation. During the process, MEG mixed with produced water and salts must be regenerated. Therefore, salt solubility and physicochemical properties of MEG aqueous mixture with electrolytes are useful to operate the regeneration unit. This work determines density and electrical conductivity data at 298.15 K for water+MEG+CaCl2 systems. The density data were correlated applying the excess volume approach and Redlich Kister expansion, resulting absolute and relative mean deviations of 3 × 10–4 g‧cm–3 and 0.028%. For electrical conductivity correlation a semiempirical model was applied resulting absolute and relative mean deviations of 1.27 mS‧cm–1 and 3.53%. CaCl2 solubilities in water-MEG mixtures at 298.15 and 323.15 K were also measured. The proposed analytical method used precise density measurements to determine salt solubility. Moreover, the solid phase composition was determined by thermogravimetry analysis. Four different solid phases were identified as function of the solvent composition. At 298.15 K the addition of MEG changed the solid phase from CaCl2•4H2O to CaCl2•3H2O•MEG for w’MEG ≥0.36. Regarding 323.15 K, solid phase changed from CaCl2•2H2O to CaCl2•H2O•MEG for w’MEG≥0.30. Density, electrical conductivities and CaCl2 solubilities were accurately determined and are of interest to the natural gas industry.

Properties of fluids under high pressure is of interest for processes design. A literature survey was conducted on experimental vapor-liquid equilibrium (VLE) and density for CO2+water and CO2+ethanol. Phase equilibrium and density of pure water, ethanol and CO2 were evaluated considering 1868 data points. VLE and saturated liquid density data were collected: 544 data points for CO2+water and 859 data points for CO2+ethanol. Soave Redlich-Kwong (SRK) equation of state (EoS) with Cubic Plus Association (CPA) was applied for calculation of these properties. Peng-Robinson EoS was evaluated to demonstrate CPA improvement in properties description. The binary interaction parameters were estimated from VLE. For pure compounds, the results indicated suitable description for both phase behavior and density. For CO2+water, density calculations were accurate (Δρ = 0.5%) and VLE was adequately described (ΔxH2O = 0.49% and ΔyCO2 = 4.63%). CO2+ethanol volumetric behavior and VLE were also satisfactorily described in CO2 concentrations up to 30% (Δρ = 2.92%, ΔxEtOH = 1.37% and ΔyCO2 = 5.86%, respectively).

Although balloon-assisted enteroscopy-endoscopic retrograde cholangiopancreatography (BAE-ERCP) is a well-described means of accessing the duodenal papilla in patients with Roux-en-Y gastric bypass (RYGB), it is associated with modest clinical success rates. Laparoscopy-assisted ERCP (LA-ERCP)-performed by advancing a standard duodenoscope through a gastrostomy into the excluded stomach and duodenum-has emerged as a viable alternative to BAE-ERCP, with apparently higher success rates. In this systematic review, we compare LA-ERCP with enteroscopy-based techniques in post-RYGB patients, including 22 case series that provided data on papilla identification, papilla cannulation, and complications. We found that LA-ERCP was superior to the enteroscopy-based techniques in its capacity to reach the duodenal papilla, although complication rates were lower for the latter. Comparative studies are needed in order to corroborate our findings.

The quality of services provided to patients in the health area is directly related to the quality of clinical information. In addition, this information must be consistent, secure and available to health professionals, even though health data is usually distributed across heterogeneous systems. The Electronic Patient Record (EPR) was proposed and applied to minimize integration problems through the construction of health information systems. This work proposes a methodology for the development of interoperable and flexible systems, using the EHRServer framework of the OpenEHR standard. As a case study, this methodology has been applied in Aracati/CE since March / 2017, in the context of the Chikungunya disease. The methodology is supported by a system that implements a set of OpenEHR archetypes representing the clinical treatment of Chikungunya. The system was tested in a Basic Health Unit. The archetypes and the MARCIA Templates were made available to the Clinical Knowledge Manager (CKM), the largest online repository of archetypes on the Web.

Os trabalhos de arqueologia desenvolvidos desde o início da última década do século XX no Centro e Nordeste de Portugal permitem estabelecer um quadro paleoambiental e cronoestratigráfico das várias fases de ocupação durante o Paleolítico Superior, bem como aperceber caraterísticas da adaptação técnica da produção das indústrias de pedra lascada, em função dos recursos líticos disponíveis localmente. Todavia, entre 40,000 e 12,000 cal BP a preservação do registo arqueológico foi fortemente condicionada pelos processos sedimentares decorrentes de bruscas mudanças nas condições climáticas globais. No Maciço de Sicó (Centro de Portugal) foram detetados artefactos e ecofactos essencialmente em cavidades cársicas e abrigos sob rocha (localmente denominados “buracas”). As sequências arqueoestratigráficas são descontínuas e terão sofrido diversas fases erosivas, que se revelam por lacunas sedimentares e superfícies de estabilização, com distintos graus de preservação. Os sítios arqueológicos da margem Norte do Rio Mondego (Centro de Portugal) estão associados a afloramentos de sílex e a sua preservação em sedimentos alúviocoluviais do Plistocénico, com uma componente eólica em depressões cársicas fechadas no Planalto de Outil/Cantanhede. No Vale do Côa (Nordeste de Portugal), os sítios ao ar livre localizam-se em antigas superfícies de erosão relacionadas com o encaixe da rede hidrográfica durante o Plistocénico Superior, assim como em depressões topográficas abertas nas superfícies aplanadas dos granitóides no limite da “Meseta”. O estudo do aprovisionamento em sílex e silcretos revela contactos de longa distância entre a Estremadura e a Meseta Norte, pelo que o mapa dos sítios do Paleolítico Superior atualmente disponível, no contexto mais geral da Península Ibérica, não constituirá uma imagem completa da densidade de ocupação humana deste território durante aquele período. Para obter uma representação mais rigorosa dessa ocupação necessitaremos inevitavelmente da aplicação sistemática de métodos de prospeção baseados em modelos de cariz geológico-geomorfológico, tal como já parcialmente estabelecido para o Valedo Côa e o Maciço de SicóOs trabalhos de arqueologia desenvolvidos desde o início da última década do século XX no Centro e Nordeste de Portugal permitem estabelecer um quadro paleoambiental e cronoestratigráfico das várias fases de ocupação durante o Paleolítico Superior, bem como aperceber caraterísticas da adaptação técnica da produção das indústrias de pedra lascada, em função dos recursos líticos disponíveis localmente. Todavia, entre 40,000 e 12,000 cal BP a preservação do registo arqueológico foi fortemente condicionada pelos processos sedimentares decorrentes de bruscas mudanças nas condições climáticas globais. No Maciço de Sicó (Centro de Portugal) foram detetados artefactos e ecofactos essencialmente em cavidades cársicas e abrigos sob rocha (localmente denominados “buracas”). As sequências arqueoestratigráficas são descontínuas e terão sofrido diversas fases erosivas, que se revelam por lacunas sedimentares e superfícies de estabilização, com distintos graus de preservação. Os sítios arqueológicos da margem Norte do Rio Mondego (Centro de Portugal) estão associados a afloramentos de sílex e a sua preservação em sedimentos alúviocoluviais do Plistocénico, com uma componente eólica em depressões cársicas fechadas no Planalto de Outil/Cantanhede. No Vale do Côa (Nordeste de Portugal), os sítios ao ar livre localizam-se em antigas superfícies de erosãorelacionadas com o encaixe da rede hidrográfica durante o Plistocénico Superior, assim como em depressões topográficas abertas nas superfícies aplanadas dos granitóides no limite da “Meseta”. O estudo do aprovisionamento em sílex e silcretos revela contactos de longa distância entre a Estremadura e a Meseta Norte, pelo que o mapa dos sítios do Paleolítico Superior atualmente disponível, no contexto mais geral da Península Ibérica, não constituirá uma imagem completa da densidade de ocupação humana deste território durante aquele período. Para obter uma representação mais rigorosa dessa ocupação necessitaremos inevitavelmente da aplicação sistemática de métodos de prospeção baseados em modelos de cariz geológico-geomorfológico, tal como já parcialmente estabelecido para o Vale do Côa e o Maciço de Sicó. Upper Palaeolithic cultures and palaeoenvironments in Central and Northern of Portugal: assessment and research prospects - The archaeological research developed during the last decade of the 20th-century in Central and Northeast Portugal established a palaeoenvironmental and chronostratigraphic framework for the Upper Palaeolithic occupation of these regions, and brought to light the operative schemes used for the production of stone tools from the locally available raw material resources. Between 40,000 and 12,000 cal BP, the preservation of the archaeological record was strongly conditioned by sedimentary processes related to rapid changes in global climate. In the Sicó Massif (Central Portugal), artefacts and ecofacts are mainly found in karst caves and rock-shelters (locally called “buracas”) whose archaeostratigraphic sequences are discontinuous, affected by erosive events (hiatuses and stabilization), and variablypreserved. The archaeological sites along the North bank of the Mondego River (Central Portugal) are associated with flint outcrops and their preservation in Pleistocene alluvium-colluvium sediments, with an aeolian component found in closed karst depressions of the Outil/Cantanhede Plateau. In the Côa Valley (Northeast of Portugal), the open-air sites are located on ancient rock-terrace surfaces related to Upper Pleistocene fluvial downcutting, as well as in topographicdepressions of the planation surfaces of the “Meseta” granitoids. The procurement of flint and silcrete reveals a large social network, extending from the Estremadura to the Northern Meseta, which indicates that, in the wider context of the Iberian Peninsula, the current distribution of Upper Palaeolithic sites is not a genuine reflection of the density of this period’s human occupation. Only the implementation of a systematic surveying approach for site detection based on geological-geomorphological models, such as those used in the Côa Valley and the Sicó Massif, will allow a more accurate reconstitution of that occupation.

Advances in cancer care delivery require revision and further development of questionnaires assessing patients' perceived quality of care. This study pre-tested the revised EORTC satisfaction with cancer care core questionnaire applicable in both the cancer inpatient and outpatient settings, and its new, outpatient-specific complementary module. The process of revision, development of the extended application, and pre-testing of these questionnaires was based on phases I to III of the "EORTC Quality of Life Group Module Development Guidelines." In phase III, patients in 11 countries in four European regions, South America and Asia completed provisional versions of the questionnaires. Fifty-seven relevant issues selected from literature reviews and input from experts were operationalized into provisional items, and subsequently translated into ten languages. Assessment of understanding, acceptability, redundancy and relevance by patients (n = 151) from oncology inpatient wards, and outpatient chemotherapy, radiotherapy and consultation settings, led to retention of, deletion of and merging of 40, 14 and 6 items respectively. Cronbach's alpha coefficients for hypothesized questionnaire scales were above 0.80. Our results provide preliminary support for the 33-item EORTC Satisfaction with cancer care core questionnaire and the 7-item complementary module specific for the outpatient care setting. A large scale phase IV cross-cultural psychometric study is now underway.

Introduction La satisfaction des soins (SdS) est un indicateur important de la qualite des soins et fait partie des « patient-reported outcomes » au meme titre que la qualite de vie relative a la sante. L’EORTC (« European organisation of research and treatment of cancer ») a developpe le questionnaire IN-PATSAT32 mesurant la SdS en cancerologie. Cet outil a ete valide specifiquement pour les patients hospitalises et ne couvre pas tous les aspects pertinents pour les patients en soins ambulatoires, une revision de ce questionnaire est donc primordiale. L’objectif de ce projet est de developper et de valider un questionnaire EORTC de mesure de la SdS, adapte a tout type de prise en charge, ainsi qu’un module specifique des soins ambulatoires. Les resultats de la phase III de validation sont presentes ici. Methodes La procedure de developpement et de validation du questionnaire suit les recommandations de l’EORTC. La phase I consistait en une revue de la litterature et entretiens semi-directifs realises aupres de 102 patients et 82 professionnels de sante issus de 10 pays, resultant en une liste de 57 items. La phase II consistait en la creation d’apres ces items d’une version provisoire d’un questionnaire central adapte a tout type de contexte de soins oncologiques hospitaliers et d’un module specifique des soins ambulatoires, construit sur une echelle de Likert a cinq modalites de reponse et traduits dans differentes langues. L’objectif de la phase III etait de realiser un pre-test de ces questionnaires afin d’identifier et de resoudre les eventuels problemes d’administration et d’identifier les items redondants ou potentiellement manquants. Cent cinquante patients etaient requis de diverses regions. La version provisoire du questionnaire composee de 57 items a ete administree aux patients ainsi qu’un questionnaire sur la pertinence et l’importance de chaque question. Un entretien structure a ensuite ete realise pour chaque patient. Les donnees sociodemographiques et cliniques des patients ont ete recueillies a l’inclusion. Les criteres de suppression des items etaient le manque de pertinence et d’importance de l’item, le taux de donnees manquantes, la moyenne de l’item superieure a 4, et un effet plafond superieur a 30 %. Des analyses descriptives ont ete realisees. Une validation preliminaire de la structure et de la fiabilite de l’echelle a ete realisee par une analyse factorielle exploratoire (AFE) et coefficient alpha de Cronbach, respectivement. Resultats Au total, 171 patients ont ete inclus pour la phase III dans 11 pays dont 71 en France (42 %) et 18 (11 %) en Grande-Bretagne. Parmi ces patients, 48 (28 %) etaient soignes en chimiotherapie de jour et 28 (16 %) en oncologie medicale. D’apres les criteres de selection et apres concertation avec les experts, 43 items ont ete retenus, 12 items ont ete supprimes, 4 items ont ete fusionnes (items 8–9 et items 20–21) et un item a ete reformule. L’instruction de reponse a ete clarifiee et une option « not applicable » a ete ajoutee pour certains items. Une comparaison a egalement ete effectuee entre les resultats obtenus pour la France et ceux des autres pays. Les resultats des AFE permettent de valider les echelles definies a priori. Le coefficient de Cronbach etait superieur au seuil de 0,70 pour chaque echelle sauf pour la dimension facilitee d’acces. Conclusion Les proprietes psychometriques du questionnaire EORTC PATSAT-C35 et de son module OUT-PATSAT8 ainsi obtenus a l’issue de la phase III vont etre analysees en phase IV.

Nowadays, there is great interest in acai fruits due to their functional and energetic properties. Based on some of these characteristics (bioactive compounds and antioxidant activity), acai fruits from different progenies were evaluated in order to select superior genetic materials. The fruits were harvested from eight progenies of the Tropical Fruits Collection at the Curu Experimental Station of the Embrapa Tropical Agroindustry, Paraipaba, CE, Brazil. Each sample was evaluated for ascorbic acid, carotenoids, yellow flavonoids, anthocyanins, total extractable polyphenols tannins and AA by the β-carotene/linoleic acid bleaching method. Regarding the majority of parameters evaluated, acai fruits presented a high variation among progenies. It was observed that the average ascorbic acid mean of the acai fruit was 58.7 mg/100 g. The anthocyanin average was 108.9 mg/100 g, with variations from 73.52 to 143.52 mg/100g, and progenies P131-3 (143.62 mg/100 g) and P034-3 (134.71 mg/100 g) showed the highest contents. In general, a high content of biological active compounds was observed when compared to others tropical fruits in the literature. Acai extracts presented an excellent oxidation inhibition, with progenies presenting different antioxidant capacities. AA varied from 47.7 to 75.0% of oxidation inhibition (% OI), with higher values for progenies P049-6 (75.0% OI), P048-8 (72.0% OI), P032-5 (70.0% OI) and P308-2 (67.0% OI).

Hyperplastic gastric polyps are often found at GI endoscopy and are not considered premalignant lesions, although some cases of malignancy have been reported. Neuroendocrine tumors, conversely, are rare and account for approximately 1% to 2% of gastric polyps. Both hyperplastic gastric polyps and neuroendocrine tumors are related to gastric atrophy. The case of a hyperplastic polyp with multifocal areas of adenocarcinoma within the polyp associated to multiple gastric neuroendocrine tumors is reported.

Disseminated microsporidiosis is diagnosed uncommonly in patients not infected with human immunodeficiency virus (HIV). We present a case of disseminated microsporidiosis in a renal transplant recipient who was seronegative for HIV. Chromotrope-based stains were positive for microsporidia in urine, stools, sputum, and conjunctival scrapings. Electron microscopy, immunofluorescence, polymerase chain reaction, and cultures of renal tissue identified the organism as Encephalitozoon cuniculi. The patient was treated with oral albendazole and topical fumagillin with clinical improvement. In addition, she underwent a transplant nephrectomy and immunosuppressive therapy was withdrawn. Follow-up samples were negative for microsporidia. However, the patient developed central nervous system manifestations and died. An autopsy brain tissue specimen demonstrated E. cuniculi by immunofluorescent staining. Disseminated microsporidiosis must be considered in the differential diagnosis of multiorgan involvement in renal allograft recipients.

Com o objetivo de selecionar pedúnculos de cajueiro-anão precoce (Anacardium occidentale L. var. nanum) para comercialização in natura, foram avaliados 09 clones selecionados a partir de um experimento de competição sob irrigação, no município de Mossoró-RN. O clone CCP 76 foi utilizado como testemunha. Os cajus foram colhidos em agosto de 1997 e avaliados quanto às seguintes características: textura, tamanho (diâmetros apical e basal e comprimento), formato, coloração (carta de cores e pigmentos) e peso (total e pedúnculo). Dentre os materiais avaliados, apenas o CCP 09 apresentou cor inferior à testemunha, sendo que os clones CAP 6 (500), END 157, END 189 e END 329 destacaram-se com coloração mais intensa que a mesma. Além da testemunha, apenas os clones END 157, 183 e 189 apresentaram pedúnculos que podem ser classificados como tipo 4 (de maior valor comercial), enquanto, com relação à forma, apenas os clones CAP 6 (500), END 157 e END 183 apresentaram formato piriforme. O clone END 157 apresentou as melhores características para comercialização in natura, inclusive quando comparado à testemunha. Os clones END 183 e 189 apresentaram resultados semelhantes à testemunha, com exceção da cor para o 183 e do formato para o 189.Nine clones of early dwarf cashew (Anacardium occidentale L. var. nanum) were evaluated within an experiment of clone competition under irrigation in Mossoró-RN, Brazil, with the objective of selecting cashew apples for fresh fruit market. Clone CCP 76 was used as a control. Cashews were harvested in august 1997 and evaluated for: firmness, size (upper and lower diameter and length), shape, color (color chart and pigment content) and weight (whole and apple). Only clone CCP 09 among those evaluated produced poorer coloring than control CCP 76, while in apples from clones CAP 6 (500), END 157, END 189 and END 329 the color was more intense than in control. Only clones END 157, END 183 and END 189 besides the control produced apples that can be classified as type 4 (highest commercial value), whilst only clones CAP 6 (500), END 157 and END 183 produced pear shaped apples. Clone END 157 presented the best characteristics for fresh fruit market even when compared to control. Results for clones END 183 and END 189 were similar to control, except for color in END 183 and shape in END 189.

A 62-year-old man was admitted to perform a colonoscopy because of chronic perianal ulcerations. These ulcers were confirmed at the perianal examination with no associated fistula. During the colonoscopy, we observed an extensive ulceration mimicking a malignancy in the ileocecal region. He referred 18 kg weight loss in the last six months and previous alcohol abuse. The physical examination showed no signs of abdominal mass. A few very small aphthous ulcers were also found in the other colonic segments. Histopathological examination and polymerase chain reaction (PCR) were applied, not confirming tuberculosis diagnosis. Another colonoscopy with biopsies was repeated, still not confirming the presence of the bacilli. In general, the tuberculosis diagnosis is made when biopsy specimens of the ileal mucosal reveal granulomas and Ziehl-Neelsen stain demonstrate acid-fast bacilli. On the other hand, the correct diagnosis was only revealed three months later, when the chest X-ray and sputum culture showed disseminated tuberculosis. Therefore, it is essential that all the specialists involved must be in agreement to reach the correct diagnosis and recommend the appropriate treatment.

Biliary endoscopic drainage using metallic self-expanded stents has become a well-established method for palliative treatment of malignant biliary obstruction. However, its occlusion, mainly by tumor overgrowth, is still the main complication without a standard treatment. We here describe a new method of treatment for biliary metallic stent occlusion, through the echo guided biliary drainage. We present a 68-year-old patient with metastatic pancreatic cancer previously treated for jaundice with ERCP and self-expandable metallic stent insertion. Four weeks later, the patient developed jaundice and symptoms of gastric outlet obstruction. A new ERCP confirmed obstruction of the second portion of the duodenum, due to diffuse tumor growth. EUS was performed, and the previous metal biliary stent was seen occluded at the distal portion in the common bile duct. A EUS-guided choledocododenostomy was performed and then, an overlapping self-expanding metal enteral stent was placed through the malignant obstruction. There were no early complications and the procedure was also clinically effective in relieving jaundice and gastric outlet obstruction symptoms. If ERCP fails in the management of occluded biliary metallic stents, EUS biliary drain can provide effective biliary decompression and should be considered an alternative to other endoscopic techniques.

A successful system of somatic embryogenesis is described for the forest tree Ocotea catharinensis Mez., which used mature zygotic embryo explants cultured on a modified Murashige and Skoog (MS) medium with activated charcoal, at 25°C in the dark. A medium composed of MS supplemented with 2% (w/v) sucrose, 0.3% (w/v) activated charcoal (AC), 362 μM 2,4-dichlorophenoxyacetic acid (2,4-d) and 0.8% (w/v) Technical Agar Grade III was used for multiplication of embryogenic cultures. Development up to the globular-stage was achieved using Lloyd and McCown woody plant medium (WPM) with 2.0% sucrose, 0.3% AC, 181 μM 2,4-d and 0.8% Technical Agar Grade III. Significant effects of media pH on differentiation of early pro-embryogenic Ocotea cell aggregates were found. Low pH of media (ca. 3–4) appeared to prevent differentiation of proembryogenic cell aggregates whereas higher pH levels (ca. 5–5.5) favoured the formation of globular structures. Once globular structures formed, they developed further to form cotyledonary somatic embryos, under the same set of culture conditions. Successful conversion of these somatic embryos to plantlets was achieved after culture on a medium composed of 1/2-strength WPM (minerals only) with 2% sucrose, 0.3% AC, 0.8% Technical Agar Grade III and 90.5 μM 2,4-d, followed by transfer to a medium composed of 1/2-strength WPM (minerals only) with 2% sucrose, 0.8% Technical Agar Grade III and 0.905 μM 2,4-d and 1.4 μM gibberellic acid, in a 16-h photoperiod regime.

Endoscopic retrograde cholangiopancreatography (ERCP) is the procedure of choice for biliary decompression in patients with unresectable pancreatic cancer. However, it may be unsuccessful in 3 to 10% of cases. When ERCP is unsuccessful, the usual alternatives are percutaneous transhepatic biliary drainage or surgery. Recently, several authors have reported the use of EUS-guided biliary drainage in patients with malignant biliary obstructions, with acceptable success and complication rates. We describe three cases of unresectable pancreatic cancer associated with obstructive jaundice, treated by EUS-guided biliary drainage.Three patients with unresectable pancreatic cancer, associated with obstructive jaundice, were included. ERCP was unsuccessful because of complete tumor obstruction of the distal common bile duct and papilla invasion. An EUS-guided rendezvous maneuver was attempted, without success. Then, EUS-guided choledochoduodenostomy, with a partially covered self-expanding metal stent, was performed in the same procedure. There were no early complications and the procedure was also clinically effective in relieving jaundice in all cases.EUS-guided biliary drainage is a feasible alternative to percutaneous transhepatic biliary drainage or surgery in unresectable pancreatic cancer with obstructive jaundice when ERCP fails. However, the development of new specific instruments and studies comparing this procedure with percutaneous transhepatic biliary drainage and surgery are needed.

Plasmacytoma presents more frequently in middle age men with aerodigestive tract involvement, especially in the head and the neck. Gastrointestinal tract involvement is uncommon, but the organ most commonly involved is the stomach. We report the first case in the literature in which final diagnosis was made by fine- needle aspiration biopsy guided by endoscopic ultrasound with adequate sample for pathologic analysis. The treatment of this entity is systemic chemotherapy but its effectiveness is limited. Plasmacytoma should be taken into account in differential diagnosis of pancreatic masses.

A 65 year-old male was admitted to the University hospital with obstructive jaundice. Endoscopic examination revealed a grossly abnormal major ampulla, situated at the edge of a large duodenal diverticulum. Biopsy of the ampulla was positive for villous adenoma. We describe the technique utilized to successfully perform an end-bloc endoscopic resection of a major ampulla on a diverticulum.

AIMS: To evaluate the overall incidence of immunoglobulin (Ig) and T cell receptor (TCR) beta and gamma gene rearrangements in a series of 40 cases of acute myeloid leukaemia (AML) and to determine whether structural modifications of these genes could be correlated with the abnormal expression of lymphoid markers in malignant cells. METHODS: All cases were classified according to the criteria of the FAB group and immunophenotyped with a panel of monoclonal antibodies reactive with myeloid and lymphoid differentiation antigens. DNA analysis was performed by the method of Southern using probes for the Ig JH, TCR-C beta 1, and TCR-J tau 1 regions. RESULTS: Phenotypic analysis showed that in addition to myeloid markers, 10 cases expressed lymphoid antigens: CD7 in seven (of which three were TdT positive, one CD2 positive, and one CD19 positive) and CD19 in three. Southern blot analysis showed that bands with sizes different from the germ line control were present in the TCR beta genes in 11 cases: in six of 30 with pure myeloid phenotype and in five of 10 of those expressing lymphoid markers. A close observation of the size and patterns of those bands, however, showed that they could be artefactual. Indeed, further analysis showed that they were either due to resistant Eco RI/Hind III sites at the beta locus or to plasmid contamination. Rearranged genes were eventually found in only two of the 40 cases: at the Ig JH region in one of the 30 with pure myeloid phenotype (3.3%) and at the TCR gamma genes in one of 10 with lymphoid markers (10%). CONCLUSIONS: These observations showed that Ig/TCR gene rearrangements were rare in this AML series (overall incidence of 5%) and that they were not significantly more common in cases with aberrant expression of lymphoid markers. The size and pattern of the potential non-germline bands that can be found in these loci must be carefully evaluated.

Proficiency testing schemes are an important tool to quality assurance in measurement as well as a tool to harmonization of multilateral recognition arrangements for accreditation. The General Coordination for Accreditation (Cgcre) of INMETRO developed a new program to accredit proficiency testing providers according with the International Standard ISO/IEC 17043. This work presents a review on needs for proficiency testing schemes in Brazil and assesses how these needs are supplied by accredited providers.

This report describes a new and improved "quick-hot Gram-chromotrope" staining technique that detects microsporidian spores in clinical specimens, such as stool, urine, saliva, nasopharyngeal fluid, and bronchoalveolar lavage samples, as well as in formalin-fixed and paraffin-embedded tissue sections.In this procedure, the samples are stained in heated (50 degrees C to 55 degrees C) solutions of crystal violet and iodine used in Gram's stain, followed by a modified chromotrope solution (heated to 50 degrees C to 55 degrees C). The modified stain is composed of chromotrope 2R (1%), fast green (0.15%), and phosphotungstic acid (0.25%).With this stain and the new protocol, microsporidian spores are stained dark violet against a pale green background, and the total staining time is shortened to 5 minutes.This new technique is fast, reliable, and simple. It can be easily adapted for use in clinical laboratories.