dataesr/scientific-paragraphs-categorization huggingface dataset:
To investigate the effects of Wolbachia groups, Wolbachia load and sampling site on CpDV load, a generalized linear model was fitted. Most significant outliers (n = 4) were removed using a multivariate approach (outlierTest, car package)  (Fox & Weisberg, 2011). We additionally analysed a second data set where we excluded all the outliers stepwise (n = 9) to verify that they did not have a major effect on our results (Figure  S2 ). The effect of a given variable was calculated as described above. The correlation between Wolbachia and CpDV loads was further calculated by linear regression. The difference between the slopes of the two wPip groups was calculated by fitting the same model with wPip × Wolbachia load interaction. The prevalence of CpDV-Ia and CpDV-Ib in wPip-I-and wPip-IV-carrying mosquitoes was compared using a χ 2 test, for the sampling sites where wPip-I and wPip-IV did not coexist (Tabarka, Ouedmelah, El battan, Ariana) versus where they both coexisted (coexistence zone; Dougga, El Manar, Fontaine, Font Mjez, Nofrancoui, Pompe, Rasrajel, Utique, Utique_Pont, Zerga) during 2010 and 2011.
| In sites where the two Wolbachia strains coexist, mosquitoes infected with wPip-IV are more likely to be infected with CpDV The absence of CpDV in some rare sampling sites (Figure  1 ) could reflect that these populations had never encountered CpDV or that the environmental conditions were not favourable for the infection. As this could bias global-scale statistical analyses, we performed the analysis on a subset of the data only considering the sites where at least one individual was CpDV-positive (per year/per site) and both Wolbachia groups coexisted. In this way, we were able to assess the possible effect of different wPip groups on CpDV prevalence under the same ecological conditions (i. e. , the similar probability of getting infected with CpDV for C. pipiens larvae harbouring wPip-I or wPip-IV).
01 to 3. 168 Wolbachia per host cell, and only marginally differed between sampling sites (df = 5, dev = 2. 476, p =. 07; Figure  3b ). In the general model, there was no significant effect of the different wPip groups on the CpDV load (glm, df = 1, dev = 0. 268, p =. 414). Similarly, larvae harbouring different Wolbachia groups did not differ significantly in terms of their Wolbachia loads (glm, df = 1, dev = 0. 007, p =. 512). However, CpDV load differed with respect to Wolbachia load (glm, df = 1, dev = 4. 072, p =. 001, Figure  3c ). We obtained similar results when we excluded all possible outliers from the data (Figure  S2 ): CpDV load was always mainly affected by sampling site (glm, df = 6, dev = 147. 32, p <. 001) and Wolbachia load (glm, df = 1, dev = 1. 523, p =. 004). To visualize the correlation between CpDV and Wolbachia loads, we also fitted simple linear regression models.
| CpDV diversification is not linked to Wolbachia groups CpDV samples were genetically diverse on the sequenced 800-bplong fragment, that is the partial NS3 protein coding region (Figure  S1 ; F I G U R E 2 Wolbachia groups and CpDV prevalence in samples from the coexistence zone. The data set was reduced only to sites where at least one individual was CpDV-positive (per year/ per site) and both Wolbachia groups coexisted. CpDV distribution was significantly affected by the Wolbachia group (glm, df = 1, dev = 16. 492, p <. 001). Mosquitoes harbouring wPip-IV were 34% (RR = 1. 338; OR = 2. 791, 95% CI: 1. 709-4. 542) more likely to be CpDV-positive. Sampling year did not have a significant effect in the model (glm, df = 1, dev = 2. 739, p =. 097). n = total sample size per wPip group/year. Lines represent 95% CI for CpDV prevalence Figure  4a ).
A total overlap between the estimated numbers of state changes (95% HPD) for both data sets indicated the lack of a phylogenetic signal linked to the wPip groups. This analysis showed that CpDV-Ia and CpDV-Ib strains were found in mosquitoes infected with either one of the wPip groups. Both tree topology together with the lack of a phylogenetic signal linked to the wPip group highlighted the horizontal transmission of CpDV between mosquito-wPip lineages. In addition to the above phylogenetic approach, we statistically compared the distribution of wPip groups and CpDV-I clades in both the sampling sites outside and inside the coexistence zone. We found that wPip groups and CpDV-I clades exhibited a parallel distribution in the sampling sites outside of the coexistence zone where only wPip-I-(Ariana, Oued Melah, El Battan) or only wPip-IV-carrying mosquitoes (Tabarka) were collected (Figure  5a ; χ 2 = 9.
67, df = 1, p =. 002). In this subset of data, CpDV-Ia was predominantly found in wPip-I-carrying mosquitoes (CpDV-Ia, n = 14, wPip-IV = 1, wPip-I = 13) while CpDV-Ib was found in wPip-IV-carrying mosquitoes (CpDV-Ib, n = 7, wPip-IV = 6, wPip-I = 1), suggesting putative local cotransmission of the two symbionts. By contrast, in the coexistence zone, the distribution of CpDV-Ia and CpDV-Ib was random with regard to their Wolbachia group infection status, i. e. , wPip-I versus wPip-IV (Figure  5b ; χ 2 = 2e-31, df = 1, p = 1) showing that horizontal transfer occurred. In line with these results, state (i. e. , carrying wPip-I or wPip-IV) changes through the CpDV phylogeny revealed a significant link between wPip group and CpDV evolution when analyses were conducted on the data set considering sampling sites where only wPip-I or wPip-IV was found (Ariana, Oued Melah, El Battan or Tabarka) (Figure  5c ) but not when performed on the data set from the coexistence zone (Figure  5d ).
(b) Total overlap between the estimated number of state (i. e. , carrying wPip-I or wPip-IV) changes for (i) the real data set (red) and (ii) the data set where wPip groups were randomized (grey) proved the lack of a phylogenetic signal linked to the wPip groups been shown to remain infective for at least a year in water bodies, although their infectivity would decrease with direct sunlight and heat  (Buchatsky, 1989). As our samples were collected in water bodies with varying ecological conditions, it was difficult to study the influence of wPip groups on CpDV in the whole area. To circumvent this limitation, and to avoid interpretation bias due to random associations, we focused our analyses on sampling sites where both wPip groups (wPip-I and wPip-IV) and CpDV coexisted, and where the larvae from each water body (i.
ACK N OWLED G EM ENTS We thank  Sandra Unal  and  Patrick Makoundou  for technical support,  Dr Philippe Clair  for his help in the qPCR experiments which were performed at the "qPCR  Haut Debit  (qPHD) Montpellier génomiX" platform. Sequencing data were produced at the  LabEX "Centre Méditerranéen de l'Environnement et de la Biodiversité " in the GENSEQ platform. We also thank  Maxime Courcelle ,  Dr Emmanuel Douzery ,  Dr Bram Vrancken ,  Dr Simon Dellicour  and  Dr Philippe Lemey  for helpful discussions. This work was funded by the  French ANR programme  (project  ANR-16-CE02-0006-01 "CIAWOL," 2017-2021 ). Sampling was done with the assistance of the  Comité de Coopération Marseille Provence Méditerranée  supervised by  Dr Robert Bismuth  in collaboration with the Tunisian ministry of ecology.
Information on the expression of the insert Protein levels of the AvHPPD-03 and PAT were analysed by enzyme-linked immunosorbent assay (ELISA) in material harvested from a field trials across four locations in Argentina during the 2011/2012 growing season. Samples analysed included leaves (V4, V8, V10, R6), root (V8, R6), forage (R6) and seeds (R8), both those treated and not treated with mesotrione and glufosinate ammonium. The mean values and ranges of protein expression levels in seeds and forage (n = 20) of the AvHPPD-03 and PAT proteins are summarised in Table  1. High variability in protein expression data is observed for all newly expressed proteins in all tissues. This variability is taken into account for the dietary exposure estimations which is discussed in Section 3. 3. 3. 1.
Consumption data Summary statistics from the EFSA Comprehensive European Food Consumption Database (EFSA consumption database) 18,19 were used as a source of consumption data when estimating dietary exposure to AvHPPD-03 and HPPD proteins. Since no specific consumption data are available for commodities containing, consisting of or obtained from soybean SYHT0H2, a conservative scenario  17 In dietary exposure assessment, different statistical estimations are used to represent high concentration when estimating worst-case scenarios (typically 95th/97. 5th percentiles). When the number of samples is lower than 60, these percentiles are not considered statistically robust (EFSA,2011), and the average of the samples above the third quartile (Q3) can be used as an alternative. 18 http://www. efsa.
europa. eu/en/data/food-consumption-data 19 Accessed on 20 December 2018 (. xlsx format). with 100% replacement of conventional soybean by the GM soybean SYHT0H2 was considered. Food commodities where proteins are expected to be absent or in negligible amounts were excluded from the assessment (e. g. soybean oil). Different processed foods (soya milk, tofu, protein concentrate and protein isolates) produced from SYHT0H2 soybean were analysed by the applicant for their content in AvHPPD-03 protein. Overall, there was a significant decrease of the levels quantified for AvHPPD-03 protein when compared to those reported in the seeds. Most probably processing conditions cause the denaturation of the proteins hampering their detection by ELISA even if the proteins might still be present in the processed foods.
Methodology Acute and chronic dietary exposure estimates for high consumers were provided for different European countries. Dietary exposure estimations were based on the methodology described for the use of the summary statistics of the EFSA consumption database  (EFSA, 2015 (EFSA, , 2019)). The assumption taken is that an individual could be a high consumer of up to two food commodities and an average consumer of the remaining food commodities containing HPPD/AvHPPD-03 proteins. Briefly, for acute and chronic dietary exposure, the two commodities leading to the highest exposure were assumed to be highly consumed (95th percentile consumption, consumers only) and to contain high and average levels, respectively, of HPPD/AvHPPD-03 proteins, while the remaining commodities (average consumption, whole population) would contain average levels of these proteins.
For the consumption of protein-based supplements, the assumption is that all of them are made of soybean protein and they all contain ~90% protein. This likely results in an overly overestimation of the dietary exposure to AvHPPD-03 protein considering that nowadays animal proteins (in particular whey proteins) are still the most commonly used protein supplements, and that protein supplements include a large array of products with diverse protein content (e. g. protein bars) and not only protein isolates with 90% protein content. Furthermore, the limited number of consumers of protein supplements in the EFSA consumption database adds uncertainty to the representativeness of the data. Together with the most likely overestimation of the dietary exposure to AvHPPD-03, there are also evidences that the current dietary exposure to native HPPD proteins is underestimated.
Bioinformatic search for homology to known toxins The AvHPPD-03 protein showed around 30% identity to bacterial haemolysins (see Section 3. 1. 2). Some bacterial HPPD proteins are annotated as haemolysins in publicly available protein databases  (Dreesen et al. , 2018)  because these catalyse the production of a haemolytic substance (a melanin-like pigment), consisting of polymers of homogentisate. This is a metabolite produced by HPPD during tyrosine catabolism under specific in vitro experimental conditions  (Dreesen et al. , 2018). The applicant provided an in vitro haemolysis study on the AvHPPD-03 protein. Under the tested experimental conditions, the protein was shown not to induce haemolysis.
It has been shown that oat can exhibit immunoglobulin E (IgE)-cross reactivity to wheat even in the absence of clinical allergy (e. g. Martens et al. , 2011). The serum samples were tested against oat extract and the recombinant E. coli derived-AvHPPD-03 protein described in Section 3. 1. 3. To determine potential IgE binding specific to the AvHPPD-03 protein, western blot with capillary electrophoresis system (WES) was used employing oat reactive sera from nine allergic individuals in one of the studies. In the other study, the Immune-CAP method was used employing oat reactive sera from 37 allergic individuals. The GMO Panel requested additional information on the Immuno-CAP-based method because several sera produced measurable levels of IgE-binding to the AvHPPD-03-loaded Immuno-CAP. The applicant provided more data on the specificity of the AvHPPD-03-binding which resulted in the finding that confounding factors could contribute to the variation in the potential IgE-binding to the recombinant E.
Conclusions The GMO Panel was asked to carry out a scientific assessment of soybean SYHT0H2 for import, processing and food and feed uses in accordance with Regulation (EC) No 1829/2003. The molecular characterisation data establish that soybean SYHT0H2 contains a single insert consisting of one copy of the avhppd-03 gene and four copies of the pat gene. Bioinformatic analyses of the sequences encoding the newly expressed proteins and other ORFs present within the insert or spanning the junctions between the insert and genomic DNA indicate a ~30% sequence identity of AvHPPD-03 to bacterial haemolysins. This finding was assessed for its relevance for food and feed safety assessment. In addition, these analyses indicate that the expression of all other ORFs showing significant similarities to allergens and toxins is highly unlikely.
The stability of the inserted DNA and of the introduced herbicide tolerance traits was confirmed over several generations. The methodology used to quantify the levels of the AvHPPD-03 and PAT proteins is considered adequate. The protein characterisation data comparing the structural, biochemical and functional properties of the plant-and microbe-produced AvHPPD-03 and PAT proteins indicate that these proteins are equivalent and the microbe-produced protein can be used in the safety studies. None of the agronomic/phenotypic and compositional differences identified between soybean SYHT0H2 and the conventional counterpart needed further assessment except for seed levels of atocopherol and c-tocopherol. These differences were further assessed for their safety and nutritional relevance and found not to raise safety concerns.
European Clinical Trials STOP! Study -Methoxyflurane vs. Placebo STOP! (ClinicalTrials. gov Identifier: NCT01420159) was a Phase 3, randomized, double blind trial comparing inhaled methoxyflurane to placebo in the ED setting. In this study, 300 patients (of whom 210 were adults ≥18 years of age) with moderate (Numeric Rating Scale [NRS] 4-7) trauma (burn, contusion, dislocation, fracture, injury due to foreign body, laceration) pain were randomized 1:1 to receive an inhaler with 3 mL of methoxyflurane or placebo. The patient could request a second inhaler, if necessary. Patients treated with methoxyflurane reported significantly greater and faster reduction in pain scores as compared to placebo: the overall difference between the methoxyflurane and placebo arms was -15. 1 mm on the VAS scale (95% CI -19.
PenASAP Study -Methoxyflurane as Part of Multi-Modal Analgesia In the French PenASAP study (ClinicalTrials. gov Identifier: NCT03798899), trauma patients with moderate to severe pain were randomized 1:1 to receive either methoxyflurane or placebo as an add-on therapy to the standard of care, as defined by the investigator. The data presented at the 2019 EUSEM congress show that the addition of methoxyflurane to standard analgesics is effective in relieving trauma pain (contusion, fracture, sprain, wound, dislocation, muscular injury, burn); the primary endpoint of median time to pain relief (defined as 30mm or less on the visual analogue scale) was 35 minutes in the methoxyflurane arm, while pain relief was not achieved in the control arm; no new safety signals were reported. While indirectly confirming that trauma pain undertreatment is still an unresolved issue in many EDs across Europe, the results of the PenASAP study establish inhaled methoxyflurane as a viable option in the context of multi-modal pain management strategies in trauma patients.
When patient and health care professional satisfaction was assessed, methoxyflurane scored higher points than the comparators in terms of pain control, comfort and ease of use. Data from the control arms of the studies that compared methoxyflurane to standard treatment as per investigator choice also showed that trauma pain is often undertreated in current clinical practice, and there is no established standard of care (e. g. , in the inMEDIATE study  29  only 23% of the patients with severe pain [>7 on the NRS scale] received opioids in the standard of care arm or as a rescue medication; in the PenASAP study  31  patients in the control arm did not achieve pain control). There are several barriers to effective trauma pain control, and they are not only complex but also generally associated with a lack of time and resources.
17 minutes severe dizziness Mercadante 2019, Phase 3b, Standard analgesic treatment Pain reduction on the VAS scale in the first No serious MEDITA (Italy) randomized, open- (per protocol) 10 minutes: -5. 94 mm (95% CI -8. 83, - adverse events NCT03585374 30 label, multicenter, 3. 06 mm; p<0. 001); time to first pain relief 9 reported active-controlled minutes Ricard-Hibon 2020, Phase 3b, Methoxyflurane or placebo as Median time to pain relief 35 minutes (not No new safety penASAP (France) randomized, open- add-on therapy to standard achieved in the control arm) signals NCT03798899 31 label, multicenter, analgesic treatment placebo-controlled (investigator's choice) Marinangeli 2018, Phase 3b, - N R N R METEORA (Italy) 32 prospective, single- arm, multicenter trial Notes: The METEORA study is currently ongoing and has not reported results yet.
The lower limit of quantification (LLoQ) in rat ocular fluids (aqueous humor and vitreous pooled together) and in rabbit aqueous humor, vitreous and retina/RPE/choroid complex was defined as 195. 3 and 39 pg/mL, respectively. Concentrations of human TF were calculated using a four-parameter logistic curve fit (4-PL, MyAssays software) from the standard curve. Values were expressed as mean ± standard error of the mean (sem) in ng/mL of fluids or ng/g of tissues.
Histology Oriented ocular globes were fixed for at least 24 h in Hartman's fixative solution (Sigma-Aldrich, St. Louis, MI, USA) and embedded in paraffin. Five-µm sagittal sections containing optic nerve (ON) were performed and stained with hematoxylin and eosin (HE). HE sections from 3 consecutive slides were examined by light microscopy at a magnification of 20× and color images were obtained using a NanoZoomer-XR Digital slide scanner (Hamamatsu Photonics, Hamamatsu city, Japan). Images were analyzed using NDP View software. The outer nuclear layer (ONL thickness) in the LID model or the total retinal thickness (from the internal limiting membrane to the retinal pigment epithelium) and the outer retinal thickness (from the outer plexiform layer to the retinal pigment epithelium) in the MNU rat model were measured every 500 µm from the ON to the inferior and the superior ciliary processes.
Louis, MI, USA) was performed to locate retinal structures. Slices were mounted with a Fluoromount mounting medium (Sigma Aldrich, St. Louis, MI, USA) under a lamella for microscopic examination using a fluorescence microscope with apotome module (Zeiss, Oberkochen, Germany), a fluorescence microscope with Spinning Disk (CSU-Xi, Intelligent Imaging Innovations, Denver, CO, USA), or a NanoZoomer-XR Digital slide scanner (Hamamatsu, Japan). Cone photoreceptors' quantification was done by masked counting for each retinal section of the number of cone-arrestin-positive cells in the outer nuclear layer. The TUNEL assay was performed on 5-µm-thick paraffin sections using a commercial apoptosis detection kit (Promega, Madison, WI, USA) and following the manufacturer's instructions. Images were recorded with a NanoZoomer-XR Digital slide scanner (Hamamatsu, Japan) at 20-fold magnification and the fluorescent staining level in the whole retina was quantified using Image J software.
Statistical Analysis To take into account the correlation between the 2 eyes of the same rat, a mixed linear model was used to define a treatment effect on ONL thickness and ERG parameters in the LID rat model. Statistical analyses were performed using SAS 9. 4 software and differences between groups and interactions were declared significant at the 5% level of significance. For the other parameters, statistical analyses were performed using Prism 8 software (GraphPad). Tests for normality for each group were performed using the Shapiro-Wilk test. For normal distribution, one-way ANOVA followed by Tukey's multiple comparisons test were used to define differences between groups. For non-normal distribution, the Kruskal-Wallis test followed by Dunn's multiple comparisons test were used to define differences between groups.
6 ± 0. 7 ng/mL in vitreous vs. 0. 31 ± 0. 2 ng/mL in aqueous humor). Six months after plasmid administration, human TF was still quantified in the vitreous of 75% of the treated eyes, allowing for a sustained expression of the protein, with an exposure and mean residence time of 184,061 pg. d. ml-1 and 58 days, respectively. Human TF was also quantified within two days in retina/RPE/choroid complex (data not shown), demonstrating that human TF produced and secreted by the ciliary muscle cells rapidly penetrated into the retina. A high TF protein concentration was observed in the retina/RPE/choroid complex 28 days following plasmid administration (Figure  2D;  3. 4 ± 1. 9 ng/g of tissue) and was still high at 6 months in this compartment (2. 5 ± 1. 3 ng/g of tissue vs. 0. 7 ± 0. 4 and 0. 1 ± 0.
6 ± 0. 7 ng/mL in vitreous vs. 0. 31 ± 0. 2 ng/mL in aqueous humor). Six months after plasmid administration, human TF was still quantified in the vitreous of 75% of the treated eyes, allowing for a sustained expression of the protein, with an exposure and mean residence time of 184,061 pg. d. ml-1 and 58 days, respectively. Human TF was also quantified within two days in retina/RPE/choroid complex (data not shown), demonstrating that human TF produced and secreted by the ciliary muscle cells rapidly penetrated into the retina. A high TF protein concentration was observed in the retina/RPE/choroid complex 28 days following plasmid administration (Figure  2D ; 3. 4 ± 1. 9 ng/g of tissue) and was still high at 6 months in this compartment (2. 5 ± 1. 3 ng/g of tissue vs. 0. 7 ± 0. 4 and 0. 1 ± 0.
0001) and in the superior retina (+33. 7%, p < 0. 0001). Despite the progression of the disease, the ONL thickness remained significantly thicker at PND60 in pEYS611-treated RCS animals in comparison to untreated RCS rats (p < 0. 0001). Cone arrestin staining and positive cells quantification were performed at PND60, i. e. , 42 days following pEYS611 ciliary muscle administration. Noticeable changes in the morphology of the remaining cone photoreceptors were observed in untreated RCS rats (Figure  9C ). Most of the outer segments (OSs) were lost and the remaining inner segments (ISs) were swollen. Similarly, the remaining terminal pedicles were smaller and flatter, with most of them directly abutting cone cell bodies, especially in the central retina (data not shown). In addition, the total number of cone photoreceptors appeared dramatically reduced compared to non-dystrophic RDY animals (16.