Quantifiable urine glyphosate levels detected in 99% of the French population, with higher values in men, in younger people, and in farmers

France is the first pesticide-consuming country in Europe. Glyphosate is the most used pesticide worldwide and glyphosate is detected in the general population of industrialized countries, with higher levels found in farmers and children. Little data was available concerning exposure in France. Our objective was to determine glyphosate levels in the French general population and to search for an association with seasons, biological features, lifestyle status, dietary habits, and occupational exposure. This study includes 6848 participants recruited between 2018 and 2020. Associated data include age, gender, location, employment status, and dietary information. Glyphosate was quantified by a single laboratory in first-void urine samples using ELISA. Our results support a general contamination of the French population, with glyphosate quantifiable in 99.8% of urine samples with a mean of 1.19 ng/ml + / − 0.84 after adjustment to body mass index (BMI). We confirm higher glyphosate levels in men and children. Our results support glyphosate contamination through food and water intake, as lower glyphosate levels are associated with dominant organic food intake and filtered water. Higher occupational exposure is confirmed in farmers and farmers working in wine-growing environment. Thus, our present results show a general contamination of the French population with glyphosate, and further contribute to the description of a widespread contamination in industrialized countries.

A systematic assessment of preclinical multilaboratory studies and a comparison to single laboratory studies

Multicentric approaches are widely used in clinical trials to assess generalizability of findings, however they are novel in preclinical experimentation. We synthesized characteristics of multilaboratory studies and quantitatively compared them to single laboratory studies. We systematically identified sixteen in vivo interventional multilaboratory studies and matched them to 100 single laboratory studies by intervention and disease. Differences in standardized mean differences (DSMD) were calculated to compare treatment effects based on study design. The multilaboratory study design was applied across a range of diseases (e.g. stroke, diabetes, trauma). The median number of labs was 4 (range 2-6) and the median sample size was 111 (range 23-384). Multilaboratory studies adhered to practices that reduce risk of bias and were transparently reported. These studies demonstrated significantly smaller treatment effects than single lab studies (DSMD 0.72 [95% confidence interval 0.43-1]). Preclinical multilaboratory studies demonstrate trends that have been well recognized in clinical research (i.e. smaller treatment effects with greater rigour in study design). This approach may provide a method to robustly assess interventions and reproducibility of findings between laboratories.

Improving replicability using interaction with laboratories: a multi-lab experimental assessment

Phenotyping inbred and genetically-engineered mouse lines has become a central strategy for discovering mammalian gene function and evaluating pharmacological treatment. Yet the utility of any findings critically depends on their replicability in other laboratories. In previous publications we proposed a statistical approach for estimating the inter-laboratory replicability of novel discoveries in a single laboratory, and demonstrated that previous phenotyping results from multi-lab databases can be used to derive a Genotype-by-Lab (GxL) adjustment factor to ensure the replicability of single-lab results, for similarly measured phenotypes, even before making the effort of replicating the new finding in additional laboratories.The demonstration above, however, still raised several important questions that could only be answered by an additional large-scale prospective experiment: Does GxL-adjustment works in single-lab experiments that were not intended to be standardized across laboratories? With genotypes that were not included in the previous experiments? And can it be used to adjust the results of pharmacological treatment experiments? We replicated results from five studies in the Mouse Phenome Database (MPD), in three behavioral tests, across three laboratories, offering 212 comparisons including 60 involving a pharmacological treatment: 18 mg/kg/day fluoxetine. In addition, we define and use a dimensionless GxL factor, derived from dividing the GxL variance by the standard deviation between animals within groups, as the more robust vehicle to transfer the adjustment from the multi-lab analysis to very different labs and genotypes.For genotype comparisons, GxL-adjustment reduced the rate of non-replicable discoveries from 60% to 12%, for the price of reducing the power to make replicable discoveries from 87% to 66%. Another way to look at these results is noting that the adjustment could have prevented 23 failures to replicate, for the price of missing only three replicated ones. The tools and data needed for deployment of this method across other mouse experiments are publicly available in the Mouse Phenome Database.Our results further point at some phenotypes as more prone to produce non-replicable results, while others, known to be more difficult to measure, are as likely to produce replicable results (once adjusted) as the physiological body weight is.

Prevalence of low alkaline phosphatase activity in laboratory assessment: Is hypophosphatasia an underdiagnosed disease?

Background Tissue-nonspecific alkaline phosphatase (TNSALP) encoded by the ALPL gene is of particular importance for bone mineralization. Mutation in the ALPL gene can lead to persistent low ALP activity resulting in the rare disease Hypophosphatasia (HPP) that is characterized by disturbed bone and dental mineralization. While severe forms are extremely rare with an estimated prevalence of 1/100.000, recent studies suggest that moderate form caused by heterozygous mutations are much more frequent with an estimated prevalence of 1/508. The purpose of this study was to estimate the prevalence of low AP levels in the population based on laboratory measurements. Methods In this study, the prevalence of low AP activity and elevated pyridoxal-5-phosphate (PLP) levels was analyzed in 6.918.126 measurements from 2011 to 2016 at a single laboratory in northern Germany. Only laboratory values of subjects older than 18 years of age were included. Only the first measurement was included, all repeated values were excluded. Results In total, 8.46% of the measurements of a total of 6.918.126 values showed a value < 30 U/L. 0.59% of the subjects with an ALP activity below 30 U/L had an additional PLP measurement. Here, 6.09% showed elevated pyridoxal-5-phosphate (PLP) levels. This suggest that 0.52% (1:194) of subjects show laboratory signs of HPP. Conclusion These data support the genetic estimation that the prevalence of moderate forms of HPP may be significantly higher than expected. Based on these data, we recommend automatically measurement of PLP in the case of low ALP activity and a notification to the ordering physician that HPP should be included in the differential diagnosis and further exploration is recommended.

Monitoring the Emergence of Algal Toxins in Shellfish: First Report on Detection of Brevetoxins in French Mediterranean Mussels

In France, four groups of lipophilic toxins are currently regulated: okadaic acid/dinophysistoxins, pectenotoxins, yessotoxins and azaspiracids. However, many other families of toxins exist, which can be emerging toxins. Emerging toxins include both toxins recently detected in a specific area of France but not regulated yet (e.g., cyclic imines, ovatoxins) or toxins only detected outside of France (e.g., brevetoxins). To anticipate the introduction to France of these emerging toxins, a monitoring program called EMERGTOX was set up along the French coasts in 2018. The single-laboratory validation of this approach was performed according to the NF V03-110 guidelines by building an accuracy profile. Our specific, reliable and sensitive approach allowed us to detect brevetoxins (BTX-2 and/or BTX-3) in addition to the lipophilic toxins already regulated in France. Brevetoxins were detected for the first time in French Mediterranean mussels (Diana Lagoon, Corsica) in autumn 2018, and regularly every year since during the same seasons (autumn, winter). The maximum content found was 345 µg (BTX-2 + BTX-3)/kg in mussel digestive glands in November 2020. None were detected in oysters sampled at the same site. In addition, a retroactive analysis of preserved mussels demonstrated the presence of BTX-3 in mussels from the same site sampled in November 2015. The detection of BTX could be related to the presence in situ at the same period of four Karenia species and two raphidophytes, which all could be potential producers of these toxins. Further investigations are necessary to understand the origin of these toxins.

ManyDogs 1: A Multi-Lab Replication Study of Dogs' Pointing Comprehension

To promote collaboration across canine science, address reproducibility issues, and advance open science practices within animal cognition, we have launched the ManyDogs consortium, modeled on similar ManyX projects in other fields. We aimed to create a collaborative network that (a) uses large, diverse samples to investigate and replicate findings, (b) promotes open science practices of preregistering hypotheses, methods, and analysis plans, (c) investigates the influence of differences across populations and breeds, and (d) examines how different research methods and testing environments influence the robustness of results. Our first study combines a phenomenon that appears to be highly robust, dogs’ ability to follow human pointing, with a question that remains controversial: do dogs interpret pointing as an informative gesture, as an imperative command, or as a simple associative cue? We collected preliminary data (N = 61) from a single laboratory on two conditions of a 2-alternative object choice task: (1) Ostensive (experimenter pointed to a baited cup after making eye-contact and saying the dog’s name); (2) Non-ostensive (experimenter pointed to a baited cup without making eye-contact or saying the dog’s name). Dogs followed the ostensive point, but not the non-ostensive point, significantly more often than expected by chance. Preliminary results also provided suggestive evidence for variability in point-following across dog breeds. The next phase is the global participation stage of the project. We propose to replicate this protocol in a large and diverse sample of research sites, simultaneously assessing replicability between labs and further investigating the question of dogs’ point-following comprehension.

The ramp and all-out exercise test to determine critical power: validity and robustness to manipulations in body position

Purpose The purpose of the present study was to determine whether a contiguous ramp and all-out exercise test could accurately determine critical power (CP) in a single laboratory visit during both upright and supine cycle exercise. Methods Healthy males completed maximal ramp-incremental exercise on a cycle ergometer in the upright (n = 15) and supine positions (n = 8), with task failure immediately followed by a 3-min all-out phase for determination of end-test power (EP). On separate days, participants undertook four constant-power tests in either the upright or supine positions with the limit of tolerance ranging from ~ 2 to 15 min for determination of CP. Results During upright exercise, EP was highly correlated with (R2 = 0.93, P < 0.001) and not different from CP (CP = 221 ± 40 W vs. EP = 226 ± 46 W, P = 0.085, 95% limits of agreement − 30, 19 W). During supine exercise, EP was also highly correlated with (R2 = 0.94, P < 0.001) and not different from CP (CP = 140 ± 42 W vs. EP = 136 ± 40 W, P = 0.293, 95% limits of agreement − 16, 24 W). Conclusion The present data suggest that EP derived from a contiguous ramp all-out exercise test is not different from the gold-standard method of CP determination during both upright and supine cycle exercise when assessed at the group level. However, the wide limits of agreement observed within the present study suggest that EP and CP should not be used interchangeably.

Single- and Multiple-laboratory Validation of LC-MS/MS Method for Simultaneous Determination of Fosetyl-Al and Phosphonic Acid in Cereal Grains and Analysis of Rice, Wheat and Barley

Background and objective In Japan, the residue definition for fosetyl-Al was “sum of fosetyl-Al and phosphonic acid expressed as fosetyl-Al” and its current provisional MRL in cereals was under review. For establishment and enforcement of fosetyl-Al MRL in cereals, a new analytical method for fosetyl-Al and phosphonic acid in cereals should be developed and validated. Methods The new method involved water extraction, clean-up using tandem cation- and anion-exchange mini-columns, and determination by LC-MS/MS. It was validated in single laboratory and multiple laboratories. Using the method, 41 samples of rice, wheat and barley were analyzed. Results In the multiple-laboratory validation: repeatability and reproducibility for three concentrations of fosetyl-Al and phosphonic acid were in ranges of 4.8–20% and 5.9–34%; calculated sum of fosetyl-Al and phosphonic acid, expressed as fosetyl-Al, showed good recoveries; linearity was observed for fosetyl-Al and phosphonic acid in ranges of 0.005–0.4 and 0.025–2.0 mg/kg; and specificity was sufficient. The method was verified for rice matrices. In 41 samples, phosphonic acid was detected up to 0.2 mg/kg while fosetyl was not. Conclusion The method was successfully validated with high precision, accuracy, linearity, and specificity and capable of analyzing fosetyl-Al and phosphonic acid with the practical LOQ of 0.01 and 0.05 mg/kg. The LOQs and concentrations of phosphonic acid in samples indicate that a potential MRL would be 0.5 mg/kg for fosetyl-Al in cereals. Highlights The validated method was simpler than many methods and did not require derivatization or matrix matched or isotopically labeled internal standards.