Seeking the driver in tumours with apparent normal molecular profile on comparative genomic hybridization and targeted gene panel sequencing: what is the added value of whole exome sequencing?

BACKGROUND: Molecular tumour profiling technologies have become increasingly important in the era of precision medicine, but their routine use is limited by their accessibility, cost, and tumour material availability. It is therefore crucial to assess their relative added value to optimize the sequence and combination of such technologies. PATIENTS AND METHODS: Within the MOSCATO-01 trial, we investigated the added value of whole exome sequencing (WES) in patients that did not present any molecular abnormality on array comparative genomic hybridization (aCGH) and targeted gene panel sequencing (TGPS) using cancer specific panels. The pathogenicity potential and actionability of mutations detected on WES was assessed. RESULTS: Among 420 patients enrolled between December 2011 and December 2013, 283 (67%) patients were analysed for both TGPS and aCGH. The tumour sample of 25 (8.8%) of them presented a flat (or low-dynamic) aCGH profile and no pathogenic mutation on TGPS. We selected the first eligible 10 samples-corresponding to a heterogeneous cohort of different tumour types-to perform WES. This allowed identifying eight mutations of interest in two patients: FGFR3, PDGFRB, and CREBBP missense single-nucleotide variants (SNVs) in an urothelial carcinoma; FGFR2, FBXW7, TP53, and MLH1 missense SNVs as well as an ATM frameshift mutation in a squamous cell carcinoma of the tongue. The FGFR3 alteration had been previously described as an actionable activating mutation and might have resulted in treatment by an FGFR inhibitor. CREBBP and ATM alterations might also have suggested a therapeutic orientation towards epigenetic modifiers and ataxia-telangectasia and Rad3-related inhibitors, respectively. CONCLUSION: The therapeutic added value of performing WES on tumour samples that do not harbour any genetic abnormality on TGPS and aCGH might be limited and variable according to the histotype. Alternative techniques, including RNASeq and methylome analysis, might be more informative in selected cases.

The fatal injuries of car drivers.

We often refer to road fatalities without knowing exactly what injuries are responsible for them. Based on the Rhône Road Trauma Registry this paper sets out to describe the topography, nature and frequency of the fatal injuries sustained by car drivers. Mean annual mortality at the wheel of a car, computed by dividing the total number of drivers killed (n=383) by the population of the Rhône Département (1.6 million) during the period 1996-2004 was 5.41 males per 100,000 and 1.41 females per 100,000, with 78% of the casualties residing in the Département. A reduction has been observed since 2003. Three-quarters of the casualties died at the scene of the crash. The results confirm the effectiveness of seat belts. The observed lethality was 0.43% for unbelted drivers and 2.7% for belted drivers (RR=0.16 [0.12; 0.21]). The injuries were analyzed for the 287 killed drivers whose deaths could be explained by the described injuries (at least one AIS 4+ injury). Of these, 41% had an ISS of 75 (at least one AIS 6 injury), 21% had an ISS of between 40 and 74, 33% an ISS of between 25 and 40, and 6% an ISS of between 16 and 24. In the case of all the AIS 4+ injuries, the three most frequent locations for injuries were the thorax only (30% of casualties), the head only (23%) and a combination of the two (18%). Abdominal injuries occurred in only 10% of casualties and spinal injuries in 9% of casualties. In the thorax, the most common injury was flail chest with haemothorax or pneumothorax. In the case of the head, the most frequent injuries were to the brain (haemorrhage, haematoma and axonal injuries). Complex fractures of the base of the skull were the second most common craniocephalic injuries. In spite of the use of restraint devices, the thorax and head are still the priority vital areas for protection in the case of car drivers. For one in four of the fatalities, death cannot be explained by any of the injuries we know about. As road traffic accidents are considered to be a cause of death in their own right, autopsies are rarely performed in France on such fatalities. This means we do not know about any injuries that cannot be detected by an external examination of the cadaver.

Whiplash risk estimation based on linked hospital-police road crash data from France and Spain.

OBJECTIVES: To investigate potential risk factors for whiplash injury as a function of crash configuration and driver's characteristics, and to provide information on over-reporting and under-reporting of whiplash. DESIGN: A case-control study of drivers involved in two-car injury collisions. Cases were drivers who had a diagnosis of whiplash injury, with or without another injury. Controls were drivers without diagnosed whiplash injury. SETTING: Hospital registries linked to police crash databases for Barcelona (Spain) and the "Département du Rhône" (France). MAIN OUTCOME MEASURES: Relative risks of whiplash and 95% confidence intervals were estimated using a modified Poisson regression. RESULTS: Of the 8720 drivers involved in car-to-car crashes recorded in the French database, 12.2% were diagnosed with whiplash; the corresponding figure in the Spanish database was 12.0% of 7558 drivers. Female drivers and drivers in rear-impact collisions were most likely to have a whiplash diagnosis, although the absolute number of whiplash cases was greater in front and side impacts. Wearing a seatbelt, being in a heavier car, and age greater than 65 years were associated with a lower risk of whiplash injury. Drivers with other injuries were also more often diagnosed as having a whiplash injury, except the most severely injured. CONCLUSIONS: Devices aimed at reducing the occurrence of whiplash injuries, such as dynamic headrest systems, should be adapted to the characteristics of at-risk occupants, especially women, and should address the mechanics of front and side impacts in addition to rear impacts.

Road crash involvement and professional status: a prospective study using the French Gazel cohort.

OBJECTIVES: To assess professional status (PS) differences in the risk of road crash involvement (RCI) (irrespective of crash severity), and to examine the underlying mechanism by evaluating the role of exposure to road risk (ERR). METHOD: A total of 15,271 subjects selected from the French GAZEL cohort were studied. A proportional hazard model for recurrent events was used to calculate the relative risks (RR) of RCI associated with PS. The associations between RCI and PS were investigated by adjusting for ERR (kilometers travelled and risk behaviors on the road). RESULTS: In all, 1890 RCI were reported. Managers have greater crude RCI risk than unskilled workers (male, RR=1.30; female, RR=1.44). This difference was no longer statistically significant when adjusting for factors describing the drivers' behaviors. Female managers' risks were also insignificant when adjusted for vehicle kilometers travelled (VKT). Managers seemed at lower risk of injury when involved in a crash. CONCLUSION: Socially advantaged subjects have the greatest RCI risk. Qualitative and quantitative ERR factors explain these disparities. These results highlight the importance to focus on ERR when studying the effect of an individual characteristic on RCI. They also highlight the importance to analyse separately the "RCI" and the "susceptibility to injury".

The recent dramatic decline in road mortality in France: how drivers' attitudes towards road traffic safety changed between 2001 and 2004 in the GAZEL cohort.

A very significant decline in the number of road casualties has been observed recently in France, concomitantly with a dramatic increase in law enforcement. The aim of this study was (i) to assess changes in attitudes about road traffic accident (RTA) prevention initiatives in France from 2001 to 2004 and (ii) to identify factors associated with an increase in positive attitudes towards RTA prevention initiatives. In 2001 and 2004, 9216 participants reported their attitudes towards traffic safety using the same self-administered Driving Behaviour and Road Safety Questionnaire. Sociodemographic, psychological and behavioural data were also available. The mean change in scores analysis showed that support for relaxing existing regulations decreased significantly during this period, while support for heightened enforcement and stricter regulations showed some decrease but remained high overall, especially concerning blood alcohol content and speed controls. Multivariate analyses suggest that highly educated drivers changed their attitudes towards road safety regulations more than other categories. Our results suggest that increased traffic law enforcement measures led to increasing support for current restrictions. Even if support for additional traffic law enforcement began to wane slightly in 2004, a large part of our population remained in favour of strengthening law enforcement related to speeding and drunk driving.

[Consequences for children involved in a road traffic accident: one year of observation and follow-up in the Rhone county]. / Consequénces des accidents de la circulation chez les enfants: suivi pendant un an dans le département du Rhône.

Children aged 6 to 11 who have been injured in a traffic accident were observed over a one year period, in parallel with a control group of children. More than one-third of the injured children had been hospitalized (for periods ranging anywhere between 1 and 47 days). One year later, one injured child out of ten was still suffering from pain, and/or still being treated for injuries resulting from the accident. Many other factors were linked to the initial overall level of severity of the injuries, contrary to that of pain, such as the rate and duration of hospitalization, the duration of care provided, the number of medical consultations, and absenteeism from school. Children who had been injured in a road traffic accident were found to be more anxious and nervous, in general, as well as having a high prevalence of sleeping disorders in comparison to the children in the control group.

Are restrained children under 15 years of age in cars as effectively protected as adults?

AIM: To compare the injury distribution between children and adults, injured as restrained car passengers. METHODS: Population based study of data from a French road trauma registry in 1996-2002. Children under 15 years old were compared with adult casualties according to the distribution of serious injuries in three distinct body regions (head, chest, and abdomen) when they were restrained car passengers. A multivariate logistic regression was performed to quantify the risk of AIS2+ injury (Abbreviated Injury Scale of 2 or more). RESULTS: Among the 7568 casualties who were injured as restrained car passengers in car accidents, 1033 were less than 15 years old. Overall, 35.4% of children and 25.2% of adults were unrestrained. For children and adults, the risk of fatality was significantly reduced when they were restrained, but the percentages of children with Injury Severity Score (ISS) > or =16, were not significantly different between restrained and not restrained casualties. Compared to adults, restrained children aged 5-9 were 2.7 times (OR 2.74; 95% CI 1.17 to 6.43) as likely to sustain an AIS2+ abdominal injury, and tended to be more at risk of AIS2+ head injuries, but were less at risk of AIS2+ chest injuries. CONCLUSIONS: Children aged 5-9 years injured in road accidents as restrained car passengers were more likely to sustain an AIS2+ abdominal injury than adults. This emphasises the need to reinforce educational campaigns aimed not only at getting children into restraint systems, but also insisting on their correct use.

Road mobility and the risk of road traffic accident as a driver. The impact of medical conditions and life events.

STUDY OBJECTIVE: We conducted a longitudinal investigation of the impact of self-reported life events and medical conditions on changes in road mobility on the wheel between 2000 and 2002 in order to assess whether these changes would affect the risk of road traffic accident (RTA). METHODS: Data are from a cohort of workers and retirees from the French national gas and electricity companies (the Gazel cohort). In the present study, 10,483 participants were included (7843 men aged 51-61 years and 2640 women aged 46-61 years, in 2000). The link between mobility and the risk of RTA was approximated using data on RTA number during lifetime and reported mobility in 2000. We then compared changes in road mobility between 2000 and 2002 resulting from life events and medical conditions reported to have occurred in the year 2001 or changed when compared to year 2000. We also compared road mobilities in 2000 in order to assess any pre-existing differences before life events and medical conditions. This led to estimation of the effect of road mobility changes on the risk of RTA. RESULTS: Changes in road mobility associated with life events and medical conditions were only found among men. These changes in road mobility were minimal. Ensuing changes in the risk of RTA were estimated to be small (odds-ratios ranged from 0.94 to 1.01). The only life events found to be associated with increased road mobility was an important purchase. Hospitalization, serious RTA, and retiring were associated with reduced road mobility. Concerning medical conditions, men who reported cataract, angina pectoris, diabetes, anxiety and stress, sleep disorder, and depression decreased their road mobility. CONCLUSION: We found no or moderate changes in road mobility resulting from life events and medical conditions, suggesting that results from previous published studies that assessed the impact of life events or medical conditions on RTA were not jeopardized by improper adjustment for road mobility.

An iterative method for calculating gamma-ray build-up factors in multi-layer shields.

Point kernel codes that simulate gamma-ray transport often use build-up factors to take scattered photons into account. This study introduces a new method, for computing multi-layer shield build-up factors. This method, based on an empirical formula for calculating double-layer shield build-up factors, is iterative. For an N-layer shield, each iteration of the method treats the first and the second layer of the shield. It replaces these layers by a single equivalent layer composed of an appropriate material and, hence, it turns the N-layer shield into an (N - 1)-layer shield. In order to determine the equivalent layer of an appropriate material, a neural network approach is developed: some neural networks trained on a large set of various configurations provide the equivalent material for any double-layer configuration. The method is implemented into MERCURE-6.3 straight-line attenuation code and is validated by comparison between MERCURE-6.3 results and reference data for one-dimensional geometries. Reference data obtained from transport calculations performed using the Sn transport code TWODANT. The comparisons prove the accuracy and sturdiness of the method.

[Differences between males and females in traffic accident risk in France]. / Différences entre les hommes et les femmes face au risque routier.

BACKGROUND: In France 7,720 people were killed in traffic accidents in 2001, 75% of which were men; the number of injured people is estimated at 153,945, of which 65% were men. The objective of the study is to describe differences between males and females regarding accidents, and to explain the main reasons for these differences. METHODS: Analysis is conducted from both national police data (2001) and data from the Rhone medical road accident trauma Register (1996-2001). RESULTS: The male/female incidence rate is 3.1 for mortality (95% CI: 3.0-3.3) and 1.7 for morbidity (95% CI: 1.7-1.8). Two-wheel motorised vehicle accidents are very specific to males, which explains part of this overrepresentation. The fatality rate and the severe injuries rate among survivors are higher for males. This is true for every main user group (car users, motorised two-wheelers, cyclists, pedestrians) after adjusting for accident circumstances and age of casualties. Males are more severely injured for all body regions and have more often severe after-effects. CONCLUSIONS: This paper shows the mechanisms leading to this unfavourable outcome for men. They correspond to differences in the number of trips, in the choice of road transport types, and moreover to differences in risk-taking behaviours. Underlying these behaviours, deep-rooted, strong and rather invariant differences between genders are to be found in the values associated with risk-taking on the road.

[Assessment of exposure to traffic pollution in epidemiological studies: a review]. / Evaluation de l'exposition à la pollution atmosphérique liée au trafic routier dans les études épidémiologiques: une revue de littérature.

BACKGROUND: Automobile exhaust is a major source of air pollution in urban areas. To study health effects of traffic exhaust fumes epidemiologists need specific tools in order to achieve a precise assessment of human exposure to traffic air pollution (TAP) and avoid misclassification. The aim of this review is to study the different ways of assessing human exposure to TAP in epidemiological studies dealing with short-term or long-term health effects of TAP. METHODS: After presenting the different designs and goals of the studies mentioned above, this review focuses on methods of assessing exposure to TAP and their different associated health endpoints. RESULTS: To assess exposure to TAP, most published studies have used more or less complex exposure indices. Several teams have used residence location and its proximity to traffic, traffic counts, or a combination of both. More recently, some authors have developed mathematical dispersion models and statistical regression models. DISCUSSION: Our analysis shows that reliable and validated tools would be needed to assess accurately human exposure to TAP. This can only be achieved with statistical regression models and mathematical dispersion models. Although such methods may be difficult to implement, their use can be facilitated by adding a geographic information system.

Traffic ticket fixing and driving behaviours in a large French working population.

STUDY OBJECTIVE: The use of connections who have the authority to cancel penalties related to traffic violations seems to be very frequent in France. This study aimed at describing risk taking driving behaviours associated in France with using connections to have traffic tickets cancelled (ticket fixing). DESIGN: Retrospective study on driving behaviour and road safety conducted between March and December 2001 within a cohort of French employees. SETTING: France. PARTICIPANTS: Employees of the French national utility company who have been followed up since 1989. The sample comprises 10 594 men aged 53-63 years and 3258 women aged 48-63 in 2001. MAIN RESULTS: One third of the men and one fifth of the women reported that they had had a ticket fixed at some time. Those who reported having tickets fixed were more likely than the others to report high driving speeds (adjusted odds ratios (aOR) were 1.24, 1.52, and 1.66 in built up areas, on rural roads, and on motorways respectively), to report driving while under the influence (aOR = 1.39), and to report risky use of cellular phone while driving (aOR = 1.83). In addition, participants who reported having tickets fixed were more likely to have had at least one serious road traffic accident in the past 11 years (aOR = 1.21). CONCLUSIONS: Indulgence and the use of connections are common practices in France. These results suggest that it is to confer a feeling of impunity that jeopardises efforts to combat unsafe driving. Abolition of these traditions is essential to ensure the credibility of preventive and repressive measures.

Traffic related air pollution and incidence of childhood asthma: results of the Vesta case-control study.

STUDY OBJECTIVE: The Vesta project aims to assess the role of traffic related air pollution in the occurrence of childhood asthma. DESIGN AND SETTING: Case-control study conducted in five French metropolitan areas between 1998 and 2000. A set of 217 pairs of matched 4 to 14 years old cases and controls were investigated. An index of lifelong exposure to traffic exhausts was constructed, using retrospective information on traffic density close to all home and school addresses since birth; this index was also calculated for the 0-3 years age period to investigate the effect of early exposures. MAIN RESULTS: Adjusted on environmental tobacco smoke, personal and parental allergy, and several confounders, lifelong exposure was not associated with asthma. In contrast, associations before age of 3 were significant: odds ratios for tertiles 2 and 3 of the exposure index, relative to tertile 1, exhibited a positive trend (1.48 (95%CI = 0.7 to 3.0) and 2.28 (1.1 to 4.6)), with greater odds ratios among subjects with positive skin prick tests. CONCLUSIONS: These results suggest that traffic related pollutants might have contributed to the asthma epidemic that has taken place during the past decades among children.

Five epidemiological studies on transport and asthma: objectives, design and descriptive results.

A case-control study was conducted in five French metropolitan areas in order to assess the role of traffic-related air pollution in the occurrence of childhood asthma. This paper presents the study design and describes the distribution of key exposure variables. A set of 217 pairs of matched 4- to 14-year-old cases and controls were investigated (matching criteria: city, age, and gender). Current and past environmental smoke exposures, indoor allergens or air pollution sources, and personal and family atopy were assessed by standard questionnaires. When possible, direct measurements were done to check the validity of this information, on current data: skin prick tests, urine cotinine, house dust mites densities, personal exposures to, and home indoor concentrations of NO(x) and PM(2.5). Cumulative exposure to traffic-related pollutants was estimated through two indices: "traffic density" refers to a time-weighted average of the traffic density-to-road distance ratio for all home and school addresses of each child's life; "air pollution" index combines lifelong time-activity patterns and ambient air concentration estimates of NO(x), using an air dispersion model of traffic exhausts. Average current PM(2.5) personal exposure is 23.8 microg/m3 (SD=17.4), and average indoor concentrations=22.5 microg/m3 (18.2); corresponding values for NO(2) are 31.4 (13.9) and 36.1 (21.4) microg/m3. Average lifelong calculated exposures to traffic-related NO(x) emissions are 62.6 microg/m3 (43.1). The five cities show important contrasts of exposure to traffic pollutants. These data will allow comparison of lifelong exposures to indicators of traffic exhausts between cases and controls, including during early ages, while controlling for a host of known enhancers or precipitators of airway chronic inflammation and for possible confounders.

Exposure to traffic pollution: comparison between measurements and a model.

French researchers from the Building Scientific and Technical Center have produced a traffic-exposure index. To achieve this, they used an air pollution dispersion model that enabled them to calculate automobile pollutant concentrations in front of subjects' residences and places of work. Researchers used this model, which was tested at 27 Paris canyon street sites, and compared nitrogen oxides measurements obtained with passive samplers during a 6-wk period and calculations derived from the model. There was a highly significant correlation (r = .83) between the 2 series of values; their mean concentrations were not significantly different. The results suggested that the aforementioned model could be a useful epidemiological tool for the classification of city dwellers by present-or even cumulative exposure to automobile air pollution.

Work-related road accidents in France.

A descriptive analysis of all victims of road accidents recorded by the police in 1997 was carried out, distinguishing accidents occurring in the course of work, while commuting and for private reasons. The analysis concerned 105,816 drivers from 14 to 64 years of age; among these, 9.9% were injured during the course of work and 18.6% while commuting. The gender-ratio was maximal for accidents during the course of work (SR = 5.5) and minimal while commuting (2.1). Accidents in the course of work were generally less serious than the two other types aforementioned. Compared with the incidence rates for labourers, those of self-employed trades and services (SETS) and those of employees and executives were higher for accidents incurred during the course of work and lower for those incurred during commuting. Professional drivers had the highest risk during the course of work. Accidents while working were most serious for SETS and professional drivers. Although it was impossible to take into account the mileage travelled, this study points out the real share of work-related road accidents and shows the need to develop and adapt prevention to the groups at risk that we have identified.

[Coagulation abnormalities after total hip prosthesis (THP), a rare cause: antibiotic prophylaxis]. / Anomalies de la coagulation après prothèse totale de hanche (PTH), une cause rare: l'antibioprophylaxie.

A 87-year-old patient developed coagulation abnormality following hip surgery related to the prophylactic use of cefamandole. Cefamandole as others cephalosporins with a methyl-tetrazol-thiol lateral chain interferes with the vitamin K regeneration cycle as do oral anticoagulants. Therefore, the use of others antibiotics or systematic vitamin K1 supplementation or single dose of cefamandole is recommended for patients with renal failure or with malnutrition. Vitamin K1 supplementation is a simple method resulting in complete resolution of the coagulation disorder.

p53-dependent G2 arrest associated with a decrease in cyclins A2 and B1 levels in a human carcinoma cell line.

In vivo transfer of wild-type (wt) p53 gene via a recombinant adenovirus has been proposed to induce apoptosis and increase radiosensitivity in several human carcinoma models. In the context of combining p53 gene transfer and irradiation, we investigated the consequences of adenoviral-mediated wtp53 gene transfer on the cell cycle and radiosensitivity of a human head and neck squamous cell carcinoma line (SCC97) with a p53 mutated phenotype. We showed that ectopic expression of wtp53 in SCC97 cells resulted in a prolonged G1 arrest, associated with an increased expression of the cyclin-dependent kinase inhibitor WAF1/p21 target gene. A transient arrest in G2 but not in G1 was observed after irradiation. This G2 arrest was permanent when exponentially growing cells were transduced by Ad5CMV-p53 (RPR/INGN201) immediately after irradiation with 5 or 10 Gy. Moreover, levels of cyclins A2 and B1, which are known to regulate the G2/M transition, dramatically decreased as cells arrived in G2, whereas maximal levels of expression were observed in the absence of wtp53. In conclusion, adenoviral mediated transfer of wtp53 in irradiated SCC97 cells, which are mutated for p53, appeared to increase WAF1/p21 expression and decrease levels of the mitotic cyclins A2 and B1. These observations suggest that the G2 arrest resulted from a p53-dependent premature inactivation of the mitosis promoting factor.

The GEN.S: a fortuitous finding of a routine screening test for hereditary spherocytosis.

As part of the evaluation of the GEN.S (Coulter), we compared the Mean Corpuscular Volume (MCV) to the Mean Spherized Corpuscular Volume (MSCV) assessed during the reticulocyte count procedure under hypo-osmotic conditions. A sub-group of patients with hereditary spherocytosis (HS) was singled out: in all of them, the MSCV became smaller than the MCV. As the cell volume normally increases in red cells derived from other patients in the same conditions, we decided to further study the reason for this particular behaviour of HS red cells. Whereas normal red cells are able to undergo an osmotic expansion, the spherocytes reach a critical osmotic volume leading to cell fragmentation consistent with the decrease of MSCV. This fortuitous finding is likely to be a reliable improvement for the routine screening of HS.