Log transformation of proficiency testing data on the content of genetically modified organisms in food and feed samples: is it justified?

The outcome of proficiency tests (PTs) is influenced, among others, by the evaluation procedure chosen by the PT provider. In particular for PTs on GMO testing a log-data transformation is often applied to fit skewed data distributions into a normal distribution. The study presented here has challenged this commonly applied approach. The 56 data populations from proficiency testing rounds organised since 2010 by the European Union Reference Laboratory for Genetically Modified Food and Feed (EURL GMFF) were used to investigate the assumption of a normal distribution of reported results within a PT. Statistical evaluation of the data distributions, composed of 3178 reported results, revealed that 41 of the 56 datasets showed indeed a normal distribution. For 10 datasets, the deviation from normality was not statistically significant at the raw or log scale, indicating that the normality assumption cannot be rejected. The normality of the five remaining datasets was statistically significant after log-data transformation. These datasets, however, appeared to be multimodal as a result of technical/experimental issues with the applied methods. On the basis of the real datasets analysed herein, it is concluded that the log transformation of reported data in proficiency testing rounds is often not necessary and should be cautiously applied. It is further shown that the log-data transformation, when applied to PT results, favours the positive performance scoring for overestimated results and strongly penalises underestimated results. The evaluation of the participants' performance without prior transformation of their results may highlight rather than hide relevant underlying analytical problems and is recommended as an outcome of this study. Graphical abstract.

Ten years of proficiency testing reveals an improvement in the analytical performance of EU National Reference Laboratories for genetically modified food and feed.

National Reference Laboratories (NRLs) in the Member States of the European Union (EU) monitor the implementation of the EU legislation on the presence of genetically modified organisms (GMOs) in food and feed. The EU Reference Laboratory for GM Food and Feed (EURL GMFF) supports the harmonisation of measurement procedures and the improvement of the analytical performance of these laboratories, among others through the organisation of a proficiency testing (PT) scheme. The PT results reported over 10 years have been analysed using common criteria applied to the reported data. The outcome revealed a gradual decrease of the relative standard deviation within the sets of the reported data with time. The extent of the deviation of the results from the assigned value also diminished between 2010 and 2019. The average deviation from the assigned value was independent of the GM content in the later PT rounds but it was affected by the complexity of the test item matrix. Performance scores were calculated for all results reported by the 62 NRLs. The number of unsatisfactory performance scores obtained decreased with time. The trends observed indicate an improvement in the analytical performance and an increased harmonisation of GMO testing within the EU enforcement laboratories.

State of the art in the determination of trace elements in seawater: a worldwide proficiency test.

This manuscript presents the results of the International Measurement Evaluation Programme 40 (IMEP-40) study, a proficiency test (PT) which was organised to assess the worldwide performance of laboratories for the determination of trace elements in seawater. This PT supports the implementation of the European Union Water Framework Directive 2000/60/EC, which aims at achieving a long-term high level protection of the aquatic environment, covering lakes, ground water and coastal waters. Forty-six participants reported results. The test item was seawater containing the trace elements As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se and Zn. The trace elements in the test item were present in very low concentrations to mimic natural levels. The results of the participants were rated with z and zeta (ζ) scores in accordance with ISO 13528 and ISO 17043. The standard deviation for proficiency assessment, σ̂, was set at 25% of the respective assigned values for the 12 measured elements based on previous experience with similar PTs. The low levels of the trace elements combined with the high salt concentration of the seawater made the measurements challenging. Many laboratories were unable to detect or quantify the elements and reported "lower than X" values. The percentage of satisfactory performances (expressed as z scores) ranged from 41% (Cr, Fe) to 86% (Mo). The PT study showed that the use of proper standard methods, like ISO 17294-2, and sensitive techniques, like inductively coupled plasma mass spectrometry (ICP-MS), contributed to performing well in this PT round.

IMEP-115: determination of methylmercury in seafood by elemental mercury analysis: collaborative study.

A collaborative study IMEP-115 was organized by the European Union Reference Laboratory for Heavy Metals in Feed and Food (EURL-HM) to validate a method for the determination of methylmercury in seafood. The method was based on a liquid-liquid extraction with an organic solvent and with an aqueous cysteine solution. The final quantitation was done with an elemental mercury analyzer. Fifteen laboratories experienced in elemental mercury analyses, from 10 European countries, took part in the exercise. Five test items were selected to cover the concentration range from 0.013 to 5.12 mg/kg. All test items were reference materials certified for the methylmercury mass fraction: DOLT-4 (dogfish liver), TORT-2 (lobster hepatopancreas), SRM 2974a (mussel), SRM 1566b (oyster), and ERM CE-464 (tuna). Participants also received a bottle of ERM CE-463 (tuna) to test their analytical method before starting the collaborative study. Method validation showed adequate accuracy and acceptable precision for all test items, thus fitting its intended analytical purpose. The repeatability RSD ranged from 3.9 to 12.3%, while the reproducibility RSD ranged from 8.4 to 24.8%.

Determination of dissolved bromate in drinking water by ion chromatography and post column reaction: interlaboratory study.

A collaborative study, International Evaluation Measurement Programme-25a, was conducted in accordance with international protocols to determine the performance characteristics of an analytical method for the determination of dissolved bromate in drinking water. The method should fulfill the analytical requirements of Council Directive 98/83/EC (referred to in this work as the Drinking Water Directive; DWD). The new draft standard method under investigation is based on ion chromatography followed by post-column reaction and UV detection. The collaborating laboratories used the Draft International Organization for Standardization (ISO)/Draft International Standard (DIS) 11206 document. The existing standard method (ISO 15061:2001) is based on ion chromatography using suppressed conductivity detection, in which a preconcentration step may be required for the determination of bromate concentrations as low as 3 to 5 microg/L. The new method includes a dilution step that reduces the matrix effects, thus allowing the determination of bromate concentrations down to 0.5 microg/L. Furthermore, the method aims to minimize any potential interference of chlorite ions. The collaborative study investigated different types of drinking water, such as soft, hard, and mineral water. Other types of water, such as raw water (untreated), swimming pool water, a blank (named river water), and a bromate standard solution, were included as test samples. All test matrixes except the swimming pool water were spiked with high-purity potassium bromate to obtain bromate concentrations ranging from 1.67 to 10.0 microg/L. Swimming pool water was not spiked, as this water was incurred with bromate. Test samples were dispatched to 17 laboratories from nine different countries. Sixteen participants reported results. The repeatability RSD (RSD(r)) ranged from 1.2 to 4.1%, while the reproducibility RSD (RSDR) ranged from 2.3 to 5.9%. These precision characteristics compare favorably with those of ISO 15601. A thorough comparison of the performance characteristics is presented in this report. All method performance characteristics obtained in the frame of this collaborative study indicate that the draft ISO/DIS 11206 standard method meets the requirements set down by the DWD. It can, therefore, be considered to fit its intended analytical purpose.

Can official control laboratories quantify reliably fipronil in eggs? Evidence from a proficiency testing round.

A proficiency testing (PT) round dedicated to assess the competence of European Union (EU) National Reference Laboratories (NRLs), EU Official Control Laboratories (OCLs) and OCLs from associated countries for the determination of the insecticide fipronil in eggs is described. The content of the target analytes (sum of fipronil plus its metabolite fipronil sulfone, expressed as fipronil in the PT test material) was set around the regulated Maximum Residue Level (MRL) set by EU legislation. The PT was organised by the European Commission's (EC) Directorate General (DG) Joint Research Centre (JRC) - in agreement with the EC DG for Health and Food Safety (SANTE) - following a request by the Belgian Authorities. Eighty-six NRLs and OCLs from 22 EU Member States, Norway, Serbia and Albania participated. The test items used were two materials made of frozen liquid eggs, processed and characterised at the JRC facilities in Geel (Belgium). The majority of the participants (94%) were shown to have a satisfactory performance, expressed as z scores, thus demonstrating the analytical capability of most of the participating NRLs and OCLs to enforce the relevant EU Regulations ((EC) 396/2005 and 1127/2014). Furthermore, 93% of the participants who provided a compliance statement classified, correctly, one of the test items as non-compliant.

The feasibility of harmonizing gluten ELISA measurements.

Many publications have highlighted that routine ELISA methods do not give rise to equivalent gluten content measurement results. In this study, we assess this variation between results and its likely impact on the enforcement of the EU gluten-free legislation. This study systematically examines the feasibility of harmonizing gluten ELISA assays by the introduction of: a common extraction procedure; a common calibrator, such as a pure gluten extract and an incurred matrix material. The comparability of measurements is limited by a weak correlation between kit results caused by differences in the selectivity of the methods. This lack of correlation produces bias that cannot be corrected by using reference materials alone. The use of a common calibrator reduced the between-assay variability to some extent, but variation due to differences in selectivity of the assays was unaffected. Consensus on robust markers and their conversion to "gluten content" are required.

Proficiency test on the determination of pesticide residues in grapes with multi-residue methods.

This manuscript presents the results of the International Measurement Evaluation Programme 37 (IMEP-37) study, a proficiency test (PT) which was organised to assess the world-wide performance of food control laboratories on the determination of pesticide residues in grapes. This PT supports the implementation of Regulation (EC) No 396/2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin. Eighty-one participants reported results, forty from EU Member States and forty-one from outside the EU. The test item was a grape sample spiked with 20 selected pesticides. The results of the participants were rated with z- and zeta (ζ-) scores in accordance with ISO 13528 and ISO 17043. The standard deviation for the proficiency assessment, σˆ, of this PT was set at 25% for the 20 measured pesticides based on previous experience with similar measurands. The results reported to IMEP-37 showed that the participants performed satisfactorily, ranging from 81% (carbendazim) to 97% (azoxystrobin, penconazole, pyrimethanil) of the participating laboratories. However, only 30% of the participants managed to analyze all pesticides satisfactorily. Overall, the performance of the participants in this PT was good but there is room for improvement in the development of multi-residue methods for the simultaneous analysis of a large number of pesticides with an increased accuracy.

Feed premix: a difficult matrix for the accurate determination of trace elements - the outcome of IMEP-114 and IMEP-36.

This paper presents the outcome of two proficiency tests (IMEP-114/36) running in parallel, which focused on the determination of total As, Cd, Pb, Hg and Sn in feed premixes. Both exercises aimed to test the performance of laboratories measuring trace elements in feed, in compliance with Directive 2002/32/EC of the European Parliament and of the Council on undesirable substances in animal feed. IMEP-114 was run for the European Union National Reference Laboratories, while IMEP-36 was open to all food control laboratories that wished to participate. In total 80 laboratories from 37 countries registered to both proficiency tests, from which 75 laboratories reported results. The test material used in this exercise was commercially available feed premix, which was processed, bottled, labelled and dispatched to participants. The performance of the participating laboratories was similar in both proficiency tests and was very good in the case of total Cd and less satisfactory for total As and total Pb. For total Sn only one-third of the participants reported results, from which 33% performed satisfactorily. Thirty-four participants reported results for total Hg, although the expert laboratories stated that the mass fraction for that measurand was below the limit of detection of the method used. An evaluation of the reasons that could be at the basis of unsatisfactory results has been carried out and is presented here.

Associating portal congestive gastropathy and hepatic fibrosis in hepatosplenic mansoni schistosomiasis.

Upper digestive bleeding is one of the most serious complications of mansoni schistosomiasis, and portal congestive gastropathy (PCG) is responsible for 25-30% of the cases of bleeding instead of bleeding due to esophageal varices. This study aimed to investigate the association between PCG with parameters of portal hypertension and the intensity of periportal fibrosis assessed by ultrasonography, in patients with mansoni schistosomiasis. A prospective study was made of 71 patients whether or not they had a history of upper digestive bleeding, and who had not been previously treated for portal hypertension (splenectomy, use of beta blockers or endoscopic treatment). Patients with other liver diseases were excluded. After signing a form of consent, the patients underwent endoscopy, as well as ultrasonography of the abdomen, and hematological, biochemical and viral markers tests. Chi-square and Fischer's exact tests were used in the statistical analysis. The mean age of the 71 patients was 50 ± 14.5 years of whom 59.2% were women. 45.1% had antecedents of upper digestive bleeding. PCG was observed in 39 patients (54.9%): severe in 8.5%, and mild in 46.5%. A positive association was observed between PCG and the grade of esophageal varices (p=0.017); and the pattern of periportal fibrosis (p=0.041). A negative association was observed between PCG and red spots on the varices (p=0.024). PCG in patients with mansoni schistosomiasis not submitted to treatment for portal hypertension is associated with the sonographic pattern of hepatic fibrosis, as well as with the grade of esophageal varices.

Optimisation and validation of programmed temperature vaporization (PTV) injection in solvent vent mode for the analysis of the 15+1 EU-priority PAHs by GC-MS.

This paper presents the optimisation of a programmed temperature vaporization solvent vent (PTV-SV) injection gas chromatographic mass spectrometric (GC-MS) method for the analysis of the 15+1 EU-priority PAHs in food extracts. Three operation parameters (vent time, vent flow and vent pressure) were optimised by applying a D-optimal experimental design. Among these variables, vent time showed the highest effect on the analytical response (signal intensity) of the target PAHs. The 15+1 EU-priority PAHs were analysed in solvent solutions and in extracts of fortified sausage. In addition, blank lamb meat extracts were prepared and spiked with the target PAHs prior to GC-MS analysis. The performance of the optimised PTV-SV injection GC-MS method was scrutinised for linearity, precision, matrix effects and robustness. All parameters were found satisfactorily. Compared to PTV injection in splitless mode, the PTV-SV injection method provided an enhancement of sensitivity for all target PAHs. Especially significant was the improvement of the S/N ratios of the compounds with the highest molecular mass.

Responsabilidade civil do médico e a inversão do pensamento jurídico sobre o tipo da atividade / Liability of the physician and the turnaround of legal thinking about the kind of activity

Sempre que uma ação causar dano a outra pessoa e houver nexo causal, isto é, quando o resultado observado pela prática desta ação estiver, diretamente ou não, relacionado, caberá a obrigação de ressarcir à vítima, um valor referente ao seu dano. Esta normatização, Responsabilidade Civil, tem como fundamento o princípio da culpa, quando subjetiva (que tem necessidade de um ato ou omissão de violar o direito de uma segunda pessoa, o dano produzido por este ato, a responsabilidade de causalidade entre o ato e o dano e, finalmente, a culpa) e o princípio do risco quando objetivo (que não necessita de culpa, já que se baseia na teoria do risco, presumindo-a, independentemente de ter ou não agido com esta intenção). Enquanto Responsabilidade Civil Contratual, duas são as obrigações de um profissional: a de resultado, ou seja, a de alcançar determinado objetivo ou fim, e a de meio, ou seja, a obrigação de empregar todos os meios para consecução de seu objetivo. No Código de Proteção e Defesa do Consumidor (Lei 8.078, de 11 de setembro de 1990), a responsabilidade objetiva deve ser empregada a todos os prestadores de serviço à exceção da classe dos profissionais liberais, médicos e advogados, por realizarem suas atividades principalmente como sendo de meios e, portanto, considerada subjetiva. Em algumas situações, o Direito entende que a Responsabilidade deve ser objetiva, ou seja, quando ficar consubstanciado uma promessa de resultado. As peças publicitárias têm tido o poder de evocar estas equivocadas situações. O objetivo principal deste trabalho é mostrar, aos profissionais do Direito em geral e aos médicos em particular, quais são os fenômenos que criaram o desvio da norma. A Responsabilidade Civil Subjetiva se encontra amparada no Código Civil, em seu Art. 186 e no caput do Art. 927, enquanto a Objetiva é observada no Art. 927, § único. Desta forma, ao relacionar os textos da literatura jurídica e os dados do Conselho Regional de Medicina do Estado de São Paulo, quanto a peças publicitárias e sindicâncias instauradas sobre este sujeito, os autores pretendem demonstrar a relação causal desta nova forma de pensar.

Whenever an action causes damage to another person and there is causal nexus, that is, when the result seen by the practice of this action is, directly or not, related, the obligation will fit to repay the victim, a referring value to its damage. This normative, Civil Liability, has as bedding the beginning of the guilt, when subjective (which needs an act or omission that finally violates the right of a second person, the damage produced for this act, the responsibility of causality between the act and the damage and, the guilt) and the risk's beginning when objective (which does not need guilt, since is based on the theory of the risk, presuming it, independently to have or not acted with this intention). While Contractual Civil liability, two are the obligations of a professional: of result, that is, to reach definitive objective or end, and of way, that is, the obligation to use all the half ones for its objective achievement. In the Code of Protection and Defense of the Consumer (Law 8,078, of September 11, 1990), the objective responsibility must be used to all the service renders, except the class of liberal professionals, physicians, and lawyers, for carrying through its activities mainly as being of ways and, therefore, be considered subjective. In some situations, Law understands that the Responsibility should be objective, that is, when a result promise is consubstantiates. The advertisements have had the power to evoke these mistaken situations. The main objective of this paper is to show, to Law professionals and, in particularly, to the doctors, which are the phenomena that had created this shunting line of the norm. The Subjective Civil Liability is supported in the Civil Code, its Art.186 and the caption of the Art.927, while the Objective is observed in the Art. 927, only §. Thus, when relating legal literature texts and data from the Regional Medical Counsel of the State of São Paulo how much the advertisements and investigations restored on this citizen, the authors intend to demonstrate the causal relation of this new way of thinking.

Hypertensive portal colopathy in schistosomiasis mansoni--proposal for a classification.

Portal hypertension is a frequent complication of chronic liver disease, detected not only in schistosomiasis, but also in cirrhosis of any etiology. Vascular alterations in the colonic mucosa are a potential source for acute or chronic bleeding and have been observed in patients with portal hypertension. The purpose of this prospective study was to describe and propose a classification for the vascular alterations of portal hypertension in the colonic mucosa among patients with hepatosplenic schistosomiasis mansoni. One or more alterations of portal colopathy were observed in all patients and they were classified according to their intensity, obeying the classification proposed by the authors. Portal colopathy is an important finding in hepatosplenic schistosomiasis and might be the cause of lower gastrointestinal bleeding in patients with severe portal hypertension.

Esclerose de varizes do esôfago em pacientes esquistossomóticos / Variceal sclerosis in schistosomotic patients

To assess the therapeutic possibilities of injection sclerosis in schistosomotic portal hypertension, a 5-year prospective study was conducted in northeast Brazil, where this parasitosis is endemic. Fifty patients undergoing endoscopy for upper gastrointestinal hemorrage from rupture of esophageal varices from July through December 1981 were chosen for the study. The 32 consenting patients were submitted to injection sclerotherapy paravariceally, using ethanolamine oleate; the 18 refusing to participate were assigned to the control group. The incidence of rebleeding was 28.1% in the former and 44.5% in the latter, a difference wich was not statistically significant (Fisher's test, p = 0.017). Since sclerotherapymarkedly improved the long-term survival rate of the patients, this procedure is advocated for the treatment of esophageal varices in cases of portal hypertension due to schistosomiasis

Hypertensive portal colopathy in schistosomiasis mansoni: proposal for a classification

Portal hypertension is a frequent complication of chronic liver disease, detected not only in schistosomiasis, but also in cirrhosis of any etiology. Vascular alterations in the colonic mucosa are a potential source for acute or chronic bleeding and have been observed in patients with portal hypertension. The purpose of this prospective study was to describe and propose a classification for the vascular alterations of portal hypertension in the colonic mucosa among patients with hepatosplenic schistosomiasis mansoni. One or more alterations of portal colopathy were observed in all patients and they were classified according to their intensity, obeying the classification proposed by the authors. Portal colopathy is an important finding in hepatosplenic schistosomiasis and might be the cause of lower gastrointestinal bleeding in patients with severe portal hypertension.