dataesr/scientific-paragraphs-categorization huggingface dataset:
The stated basis for the NOAEL for βcaryophyllene epoxide was the reported erythrocyte infiltration into the sinusoids of the mesenteric lymph nodes, which could not be dismissed. However, erythrocyte infiltration into the sinusoids of the mesenteric lymph nodes was only observed at the highest intake level of β-caryophyllene epoxide, and on this basis alone the NOAEL would be equal to the middle dietary level (10,500 ppm) in this study (equivalent to 672 mg/kg bw/day in males). In addition to the standard subchronic toxicity endpoints, endpoints of male and female reproductive function were also included in both studies to generate relevant data that were lacking in the toxicological database for these flavoring or fragrance materials. The administration of either β-caryophyllene or β-caryophyllene epoxide in the diet did not influence estrus cycle pattern in females, based on mean estrus cycle length and the number of cycles assessed in two intervals during the study (Weeks 6-7 or 12-13).
Acknowledgments This work was supported by the  International Organization of the Flavor Industry  and the  Research Institute for Fragrance Materials, Inc. The authors declare that there are no conflicts of interest. The authors thank  Dr. Elke Kennepohl  for her expert contribution in drafting the original Methods and Results sections and  Drs. Xiaodong Li ,  Craig Llewellyn , and  Atish Patel  for their assistance in the preparation of this manuscript.
X-ray crystallographic data were collected on an APEXII crystal diffractometer. Specific rotation (in deg cm 3 g -1 ) was measured on a Perkin Elmer-341 polarimeter. Melting points were measured on a melting point apparatus. Stuart SMP10. 1-Ethynylcyclopent-1-ene, 35 1-ethynylcyclohex-1-ene, 36 1ethynylcyclohept-1-ene, 37 1-ethynylcyclooct-1-ene, 37 5,6dihydro-2H-pyran-3-carbaldehyde, 38  1-tosyl-1,2,3,6tetrahydropyridine-4-carbaldehyde, 39 9h, 40 10, 41 11, 34 9j, 42 9k 43 were prepared as previously described. Perillaldehyde and 3-(allyloxy)-1-propyne 9i are commercially available. General procedure for the synthesis of boronates 7a-7d by catalytic hydroboration. To a mixture of 1ethynylcycloalk-1-ene (1. 86 mmol) and Schwartz catalyst (104 mg, 0. 37 mmol) at 0 °C under argon atmosphere, was added dropwise H-Bpin (216 μL, 1.
ACKNOWLEDGMENTS We acknowledge the  Ministère de l'Education Nationale, de la Recherche et de la Technologie , the  Centre National de la Recherche Scientifique (CNRS)  and the  Région Bretagne  for financial supports. We also thank  Andrei S. Batsanov  ( Chemistry Department, Durham University ) and  Marie Cordier  ( Centre de Diffractométrie X, ISCR, Rennes ) for X-ray crystallographic structure determinations.
Development of C109 nanocrystal-embedded dry powders In depth formulation studies were devoted to the development of C109 nanocrystal-based dry powders to be reconstituted in aqueous medium for inhalation suspension. First, aqueous dispersions of drug nanocrystals were produced by a nanoprecipitation technique. Three variables were considered: drug solvent, C109 concentration and type of stabilizer. Optimal formulation parameters were chosen based on size, PDI and ζ of the nanocrystals. Of note, unreliable data were acquired by dynamic light scattering when using DMSO or THF as solvents, independently on C109 concentration (Table  S1 ). The particle size distribution, as determined by laser diffraction analysis, confirmed the micrometric dimensions of the crystals, displaying D90 ~4. 5 μm.
This observation provides further evidence of a deconstruction of the nanocrystal after long time in artificial CF mucus, consistently with SANS data. cell lines was assessed by the MTT assay. As previously shown, the toxic concentration at 50% (TC50) of C109 on these cell lines is about 75 μM. 11 C109 nanocrystals showed the same behavior as C109 compound in 16HBE cells (Figure  7 , A), while the TC50 of TPGS_C109 was greater than 100 μM in CFBE41o - cells (Figure  7, B ). The corresponding controls, TPGS and CD alone (Figure  7 ), as well as the combination of CD and TPGS (data not shown) did not cause significant toxicity. Of note, the TC50 of C109 and the corresponding TPGS_C109_CD1. 0 was at least 3-fold higher than the microbiologically active concentration of the compound on the most sensitive bacterial strains, that is 13.
The same result was not achieved when C109 compound was used in combination with piperacillin (data not shown). These observations were confirmed by measuring the bacterial load (CFU/larva) at 48 and 72 h p. i. in the hemolymph of two larvae/group: only upon treatment with piperacillin alone or the combination piperacillin plus TPGS_C109 was the bacterial load significantly reduced (data not shown).
Acknowledgments G. R. , F. U. and V. M. wish to thank the  Italian Cystic Fibrosis Research Foundation  (Project  FFC  # 19/2015  adopted by  Gruppo di Sostegno FFC di Como Dongo, Delegazioni FFC di Olbia Tempio e di Reggio Calabria ) and the  US Cystic Fibrosis Foundation  (grant  RICCAR17G0 ) for funding support. The work was supported also by a  BlueSky research grant  of the  University of Pavia  to S. B. and by the  Italian Ministry of Education, University and Research (MIUR)  ( Dipartimenti di Eccellenza ,  Program 2018-2022 ) to  Department of Biology and Biotechnology, "L. Spallanzani", University of Pavia  (to G. R. , S. B. , L. R. C. ). P. B. and V. R. were supported by research grant from  BioMeTra Department of the University of Milano. P. B.
Sponge characterisations Scanning Electron Microscopy (SEM, S-4700 field emission GU, Hitachi, Germany) was used to evaluate the morphology of the sponges. Accelerating voltage was set up at 5 kV and the emission current at 10 μA (magnification x50). Samples were chrome coated before observations. Due to the heterogeneous shape of the pores of the sponge, a size analysis was performed by measuring the diameter of 20 pores randomly selected using the software ImageJ. Swelling properties were assessed with dry cylindrical sponges (∅11 mm x 5 mm). Sponges were weighed and immersed in a PBS solution (10 mL per 20 mg of sponges) at 37 °C and 80 rpm. At regular time intervals (30 min and every hour for 6 hours with a final weighing at 24 h), the sponges were collected, dried on an absorbent paper tissue and weighed.
aureus colonisation, histology or FACS analysis as required. For determination of S. aureus colonisation, skin was collected into pre-weighted tubes containing 2 mL BHI. Tissue was homogenised and 100 mL was plated out onto selective BHI agar containing (1. 5 mg mL À1 Oxacillin, 12. 5 mg mL À1 Ampicillin, 2 mg mL À1 Polymixin B). Colonies were counted aer 24 hours at 37 C and data are expressed as cfu g À1 skin. 2. 4. 3 Flow cytometry. For ow cytometry analysis skin was collected into ice cold DMEM medium (Gibco BRL) containing 2% FCS. Lymphocytes were isolated from skin using collagenase/dispase treatment as described in. 61 Aer blocking Fc receptor sites, cell suspensions were stained with ZombieAqua (Biolegend), and then with a cocktail of surface markers: CD45, CD3, CD4, CD8, CD11b, Ly6G (all from Biolegend).
Single-stained cytometric beads were used as compensation controls and labelled non-specic isoform control antibodies as negative controls (Biolegend). Stained were cells were washed and acquired using a FACSCanto II with FACsDiva (Becton Dickinson), analysis was performed using FlowJo soware (V10. 4. 3, FLowJo Treestar, LLC). Cells were gated to exclude dead cells and aggregates, and gating was applied to detect lymphocytes based on FSC ad SSC characteristics. Inammatory cell populations were dened as follows: neutrophils (CD45 + , CD3 À , CD11b + , Ly6G + ), macrophages (CD45 + , CD3 À , CD11b + ), CD4 + T cells (CD45 + , CD3 + , CD4 + ). Statistical analysis was performed using GraphPad Prism (V8. 0. 1, GraphPad Soware Inc. ). Datasets were tested for normality (D'Agostino & Pearson test), and because data for some groups were found not to be normally distributed, groupwise comparisons to untreated controls were made using the non-parametric Kruskal-Wallis test with Dunn's correction for multiple comparisons.
Our objective was to quantify the impact on the use of zolpidem of the obligation implemented in France in 2017 to use secure prescription pads to prescribe it. We conducted a cohort study within the French SNDS healthcare database. Patients aged over 18 years of age were considered for inclusion. The number of prevalent users and incident episodes of zolpidem use were compared before the change in law (July 1, 2016 to January 1, 2017) and after (July 1, 2017 to January 1, 2018). A prevalent user was a patient who has been reimbursed for zolpidem at least once. An incident episode of zolpidem use was defined by a first administration of zolpidem without any prior administration within the previous six months. Regarding prevalence of zolpidem users, we observed a decrease from 2. 79% (CI95%:2.
Participants and outcomes The study sample included all patients in the Generalist Sample of Beneficiaries (EGB). EGB is a 1/97 th representative sampling of the SNDS. The SNDS links several existing databases: the nationwide claims database of the French National Healthcare system (Système National d'Informations Interrégimes de l'Assurance Maladie; SNIIRAM); the national hospital database (Programme de Medicalisation des Systèmes d'Information; PMSI) and the national death registry (Centre d'epidemiologie sur les causes medicales de Decès; CepiDC). The SNDS covers more than 98% of the French population (66 million people) from birth (or immigration) to death (or emigration), even in case of change in occupation or retirement. Data are individual and anonymous. The SNDS contains a longitudinal record of health encounters, hospital criteria for access to confidential data.
Data collection The target population of our research was consumers of zolpidem included in the EGB aged over 18 years old. A prevalent user was defined as a patient being reimbursed at least once for zolpidem and an incident episode of zolpidem use was defined as a patient receiving a first delivery of zolpidem without any prior deliveries over the preceding 6 months. Two time periods were used: (i) period 1 from July 1, 2016 to January 1, 2017, reflecting zolpidem consumption before the change in law (ii) period 2 from July 1, 2017 to January 1, 2018 reflecting the steady state of zolpidem consumption after the change in law. The variables gathered in the database included age at inclusion, sex, information on active CMUc (for "Couverture medicale universelle complementaire", which indicates low-income status granting full reimbursement of health expenditures), and the presence of a chronic disease via registration in the list of chronic diseases scheme beneficiaries (ALD for "Affections de Longue Duree").
Statistical analysis Prevalences were described with their point estimates and 95% accuracy intervals. The comparison of prevalences between both periods was performed with a McNemar test. Let's consider probabilities of respectively stopping treatment in period 2 when you are a consumer in period 1(p stop ) and starting treatment in period 2 when you did not consume in period 1 (p start ). The null hypothesis of the test is p stop equal p start. The alpha risk was set at 5% and the statistical significance threshold set at 5%. This analysis was done on the dataset containing only patients present in the EGB database over the 2 periods. A sensitivity analysis was conducted by recoding patients missing in a period as non-consumers in the period considered. Incidences were first described by constructing rates of treatment initiations per 100,000 insured per month (patient-month).
The description of incidences trends was then conducted through the construction of incidence rate ratios (IRRs) adjusted for the time period with a negative binomial model. In order to study effects of potential confounders of the effect of change in law, a logistic regression was conducted. The variables to be included in the model were defined a priori and included period (before vs after change in law), age in 2016, sex, being under a CMUc scheme or an ALD scheme. Age was introduced into the model in class intervals because the hypothesis of a constant effect of aging on the probability of receiving zolpidem regardless of the age stratum in which patients are located was unlikely to be respected for this variable. The adequacy of the model to the data was checked graphically. The dependencies between observations in a same patient over the two time periods were handled by the use of Generalized Estimating Equations (GEE), with the use of a robust variance estimator in order to facilitate convergence and improve the accuracy of confidence intervals.
Discussion We found that the change in law that occurred in the beginning of 2017 regarding zolpidem prescription was concomitant with the halving of the prevalence of zolpidem users. This association was independent of age, sex, from suffering from a chronic disease granting an ALD, and from being precarious enough to benefit from the CMUc scheme. We also observed that the incidence of new zolpidem treatment episodes halved after the promulgation of the law. The main strength of this study is that we have provided recent population-level data across a representative sample of the exhaustive national reimbursement database. The validity and usefulness of this database in pharmaco-epidemiological studies have been previously studied and validated, and the analysis of large medico-administrative databases offers the advantages of completeness of patients and unbiased study sample  [6].
The main limitation is the total absence of clinical information, e. g. concerning the effects searched for and felt by patients, the modification in routes of administration, the tolerance and the withdrawal. We have no information either about therapeutic strategies used to reduce zolpidem use. As we rely on a medico-administrative reimbursement database, we have no information on illegal drug use or purchase of zolpidem or other therapies. A complementary in-field study is currently ongoing to document these points  [7]. These initial results showed that the enforcement of the use of secured prescription sheets had an effect on the population's exposure to zolpidem. Since there are cues that prescription drug abuse varies with the availability of the involved medication  [8] , our results suggest that the change in law could be effective in limiting the entry of new users into abuse or dependence.
These results are concordant with observations of primary care pharmacists, albeit in a more restricted area  [9]. Among the 235 responding pharmacies in the French Rhone area, zolpidem dispensation decreased by an average of 41. 8%. On the same time windows, zopiclone dispensations for the 3. 75 and 7. 5mg dosages increased by 18. 4 and 16. 7% respectively. Our results were also concordant with another study using the French National Health Insurance database. One year after the information of the ANSM about the measure, the prevalence reimbursement for zolpidem among individuals who had at least one hypnotic drug reimbursement decreased from 26. 0% to 18. 4%, while the prevalence of reimbursement for zopiclone increased from 18. 0% to 28. 3%  [10]. https://doi. org/10. 1371/journal. pone.
As an example, recreational users may have switched to other drugs worse than zolpidem, which may have turned the measure counter-productive in this specific subpopulation. The real impact on patients suffering from zolpidem addiction is still largely unknown and SNDS data do not allow this point to be accurately documented. However, the principles of management of dependence with 'z-drugs' such as zolpidem and zopiclone are the same as the management of benzodiazepine dependence  [17]. An interesting point would be to monitor the use of other alternative therapies (e. g. zopiclone, benzodiazepines) to better understand the decrease in the use of zolpidem after the measure. Another lead could be to monitor changes in inadequate behavior, such as doctor shopping and use of excessive doses, which already proved to be useful in studying drug misuse using the SNDS database  [13, 18].
These investigations are currently conducted. All these point are crucial in order to evaluate if the change-in-law had a beneficial effect, both in terms of individual health and public health. Regarding the associations between zolpidem consumption and sex, older age, low income and presence of comorbidities, these are consistent with what has already been observed in 2015 on data issued from the same database, but analyzed by another team  [19]. These associations are also observed in other settings, which suggest that these are not specific to the French population  [20, 21]. It should be noted that zolpidem is the most frequently prescribed hypnotic drug in the elderly  [19] , for whom the risk of dependence is high  [22, 23]. Studies investigating the evolution of consumption in this specific population are needed.
Grant support and Acknowledgements: This work has been supported by funding from EU H2020 Research & Innovation programme, No. 731875 (LIVERHOPE). We acknowledge Nicola Van Berckel for her administrative support and her valuable collaboration with the development of the Liverhope Project. We acknowledge the collaboration of UICEC IBIMA, Plataforma de Unidades de Investigación Clínica y Ensayos Clínicos (SCReN) (PT13/0002/0020 and PT17 0017 0020), cofounded by  FEDER.
Treatment was started after the baseline visit and was given for twelve weeks. Study visits were performed every two weeks; standard clinical and analytical data were collected and complications of cirrhosis and treatment-related adverse events were assessed and registered, if any. Compliance with the study medication was assessed at each visit by self-reporting and confirmed by counting the pills of simvastatin and rifaximin returned by the patients at the end of the treatment period. Subjects with a compliance of less than 70% of the total supplied study medication were excluded of the per protocol analysis. Study medication was permanently withdrawn if patients developed liver or muscle toxicity. In addition, treatment was also permanently withdrawn in patients who developed HE and met the criteria for treatment with rifaximin according to the EASL guidelines  [17] , and in those patients with severe treatment-related side effects, according to the judgement of the investigator.
Randomization, blinding and treatment allocation Eligible subjects were randomized 1:1:1 to the different groups of the study. Randomization was stratified according to Child-Pugh class (B vs C) using the PROC PLAN of the SAS system and restricted using blocks multiple of 3 elements. A centralized computer-generated randomization was used through the electronic Case Report Form (eCRF). Unblinding was not expected until all subjects had completed the study and the database had been locked after data completion and verification. However, it was foreseen that a specific patient could be unblinded, if needed, as judged by the investigator in emergent cases with side effects possibly related to the study medication. Main safety outcome measures in patients included in the study were monitored by an independent Data Safety Monitoring Board (DSMB), constituted by a group of experts.
Statistical methods Categorical variables were summarized by counts and proportions and continuous variables by mean (standard deviation) as appropriate. Longitudinal continuous variables assessing the individual laboratory parameters measured to evaluate toxicity (AST, ALT, AP, and CK levels) were analysed using Mixed Models for Repeated Measurements including the Child-Pugh stratum and the baseline measurement. The Fischer's exact test was used to compare categorical variables. Missing data for longitudinal variables was handled through the MMRM strategy which relies on the missing at random assumption. Categorical variables were handled using a worst-case imputation strategy and any treatment discontinuation due to relevant safety issues was considered as a failure. Since the objective of this exploratory trial was to assess and discard safety signals, no multiplicity adjusting strategy was planned.
Therefore, the full analysis set included a total of 44 patients (16 patients in the simv40+rif group, 14 patients in the simv20+rif, and 14 patients in the placebo group). The overall median follow-up period was of 84 days (IQR 36-84). The majority of patients were compliant with the study medication (mean compliance among the three treatment arms included in the study of more than 90% of fulfillment with rifaximin/ placebo and simvastatin/ placebo). Only four patients (three in the group of simvastatin 40mg/day plus rifaximin and one in the group of simvastatin 20mg/day plus rifaximin) were excluded of the per protocol analysis for lack of compliance with the study medication. Baseline demographical, clinical and analytical data were quite similar in the 3 groups, except for lower age in the simv20+rif group and slightly higher frequency of HE and variceal bleeding in the placebo group compared to the other two groups (table  1 ).
Side effects were apparently not related to presence or frequency of polymorphisms of the gene SLCO1B1 encoding OATP1B1. Another potential explanation for the high frequency of adverse events observed in the current study could be a toxic effect of rifaximin either by itself or by increasing the toxicity of simvastatin. The rifampin component of rifaximin could theoretically induce muscle toxicity through mitochondrial oxidative stress and perhaps act synergistically with statins. This potential mechanism was raised on the basis of a case reporting a chronological relationship between treatment with rifaximin and rhabomyolysis in a patient with cirrhosis  [26]. However, the possibility of muscle toxicity due to rifaximin seems remote due the following reasons: 1/ no cases of rhabomyolisis were reported in the pivotal trial of rifaximin for prevention of recurrent of HE and its long-term followup  [15] ; 2/ as of July 2019, only two cases of rhabdomyolisis possibly related to rifaximin have been collected in postmarketing safety databases of AlfaSigma (Bologna, Italy) over more than 10 years; 3/ rhabdomyolisis has been reported in the setting of advanced cirrhosis and is idiopathic in 25 to 58% of cases  [27] ; therefore, cause-effect relationship is difficult to establish; and 4/ no muscle toxicity was found in animal studies in rats and dogs treated with doses of rifaximin equivalent to those used in humans  [28].
Nevertheless, since safety of the combination of simvastatin and rifaximin remains a relevant issue, the LIVERHOPE Efficacy Trial, a randomized double-blind trial comparing simvastatin 20 mg/day plus rifaximin vs placebo in the prevention of ACLF in patients with decompensated cirrhosis which has already started includes an assessment of safety at the time when the first 40 patients randomized to the treatment arm have reached at least one month of therapy (NCT03780673). Other aspects of the study deserve a comment. First, it should be emphasized that liver safety monitoring and stopping rules for drug-induced liver injury in patients with decompensated cirrhosis enrolled in clinical trials remain a challenge because of potentially altered liver tests before initiation of treatment  [29]. Therefore, potential liver toxicity was assessed by evaluating changes with respect to baseline.