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Additional file 1: Figure S1. Generation of polyclonal LIG4 knock-out HAP1 cells. A. Schematic diagram of the LIG4 DNA region targeted by the gRNA, and localization of the AflIII restriction site used to knock out this gene in HAP1 cells. B. Schematic diagram of the protocol used to generate the polyclonal LIG4 KO cell line. C. Electrophoresis results following digestion with the AflIII restriction enzyme for screening of the clones. D. Sanger sequence alignments of the clones selected on basis of AflIII restriction site loss relative to HAP1 parental cells. E. Sanger electrophoregrams of LIG4 KO clones with insertions or deletions causing frameshifts relative to the HAP1 parental cell line. Figure S2. Analysis of coverage and frequencies of insertions and deletions obtained following genome editing and NGS sequencing of BRCA1 and BRCA2 variants. A. Insertion and deletion frequencies determined following the NGS sequencing of BRCA1/2 variants of interest, compared with those for the corresponding silent control. These means include the 7 nt surrounding the mutations of interest. B. Analysis of NGS sequencing coverage with the following formula: Mutation of interest (Mutation+Reference control)/Silent control (Mutation+Reference control). Results for all BRCA1/2 variants characterized by comparison with the classified benign and pathogenic mutations (Mann-Whitney tests, p = 0.1934 and p = 0.2902). C. Frequencies of insertions and deletions according to the NGS sequencing of BRCA1/2 variants and corresponding silent editing controls and linear regression analysis. These means include the 8 nt surrounding the PAM sequence. Variants classified in databases are shown in colours: benign mutations in green and pathogenic mutations in red. Figure S3. Evaluation of the reproducibility of our functional assay. A. Replicates of functions scores from four different variants obtained by following the described protocol in the LIG4 KO HAP1 cells (n = 3). Among these, the variants p.Pro1812Ala and p.Tyr422X were already classified as pathogenic in databases, the other two were VUS or had a conflicting interpretation. B. Sanger electrophoregrams of XRCC4 KO clones with insertions or deletions causing frameshifts relative to the HAP1 parental cell line. Clones 5 and 17 were pooled to generate the polyclonal XRCC4 KO HAP1 cell line. C. Comparison of the functional scores evaluated after CRISPR-Cas9 editing and NGS sequencing of four BRCA1 variants in polyclonal LIG4 KO cells and polyclonal XRCC4 KO cells. Table S1. gRNA and oligonucleotides sequences designed to edit BRCA1, BRCA2 and POLE variants in the study. Table S2. Odds of Pathogenicity (OddsPath) estimated by performance of classified variant controls. In this assay, five benign and five pathogenic variants of BRCA1 and BRCA2 were used as controls allowing the estimation of the evidence strength of our classification according to Brnich et al [34] guidelines. Table S3. Variants used as silent controls or references with their classification according to different databases. Benign mutations are shown in green (+), unreported mutations are shown in yellow (?), mutations with conflicting interpretation are shown in orange (+/-), pathogenic mutations are shown in red (-) and intermediate mutations are shown in blue (I).

Introduction and Aims Concomitant elevated alcohol consumption and use of benzodiazepines (BZD) during methadone treatment is widespread and particularly worrying because of the increased risk of overdose. Using concomitant binge drinking and use of BZD as a proxy of overdose risk, we aimed to study whether buprenorphine switchers were at higher risk of overdose during methadone treatment. Design and Methods The French National Agency for Research for Aids and Viral Hepatitis –Methaville multisite randomised trial enrolled 195 patients to assess the feasibility of initiating methadone in primary care by comparing it with methadone initiation in specialised centres. We selected 174 patients with available data on BZD use and alcohol binge drinking at baseline and 12 months, accounting for 318 visits. The outcome was defined to take into account an overdose risk gradient as follows: no BZD use, BZD use without and with binge drinking during the previous month. To identify factors associated with the outcome, we performed a mixed multinomial logistic regression analysis. Results At baseline, 26% of the sample reported BZD use alone while 16% reported BZD use and binge drinking. Half of the sample (51%) was switching from buprenorphine treatment. After multivariate analysis, employment, depressive symptoms and switching treatment from buprenorphine to methadone [odds ratio (95% confidence interval) 5.38 (1.74–16.62)] remained associated with BZD use and binge drinking. Discussion and Conclusions As well as the importance of identifying socially vulnerable and depressed methadone-maintained patients, clinicians should be aware that patients who fail buprenorphine treatment and switch to methadone require greater clinical monitoring and management to avoid the risk of overdose. [Roux P, Lions C, Michel L, Mora M, Maradan G, Marcellin F, Spire B, Alain M, Patrizia CM, the ANRS Methaville Study Group. Concomitant use of benzodiazepine and alcohol in methadone-maintained patients from the ANRS Methaville trials: Preventing the risk of opioid overdose in patients who failed with buprenorphine. Drug Alcohol Rev 2015; ●●:●●–●●]

Dans la société rurale picarde les rapports de force mettent aux prises tout autant les ouvriers agricoles qui ne sont pas sûrs de trouver un travail ni de le garder, qu'adversaires de classes. Le climat de tension qui règne dans les fermes résulte tout d'abord de la forte pression exercée par un patron-fermier qui dispose d'une force de violence sociale faisant de lui le maître du sort de chacun. Il s'explique aussi par la compétition entre ouvriers disposés à se distinguer au regard d'échelles hiérarchiques fondées sur le comportement au travail et la capacité à faire plus que les autres. L'analyse du vocabulaire assez riche et précis qui désigne les différentes façons de travailler est mise en relation avec une morale de la vie quotidienne qui fait de l'effort la valeur cardinale. Social tension in Picardie. -- In the rural society of Picardie conflict relationships imply the agricultural workers who are not sure to find a job and to keep it, as well as class opponents. The atmosphere of tension which dominates in the farms is the result of the pressure exercised by the landlord-farmer who disposes of a powerful social violence, making himself the master of their fate. This can also be explained by the competition that exists between the workers able to distinguish themselves on a hierarchical scale based on behaviour at work and their ability to achieve more than the others. The analysis of their rather rich and precise vocabulary which defines the different working methods is put in relation with the morality of everyday fife which underlines the importance of effort as the main objective value.