Dissection of the Fgf8 regulatory landscape by in vivo CRISPR-editing reveals extensive intra- and inter-enhancer redundancy.

Developmental genes are often regulated by multiple elements with overlapping activity. Yet, in most cases, the relative function of those elements and their contribution to endogenous gene expression remain poorly characterized. An example of this phenomenon is that distinct sets of enhancers have been proposed to direct Fgf8 in the limb apical ectodermal ridge and the midbrain-hindbrain boundary. Using in vivo CRISPR/Cas9 genome engineering, we functionally dissect this complex regulatory ensemble and demonstrate two distinct regulatory logics. In the apical ectodermal ridge, the control of Fgf8 expression appears distributed between different enhancers. In contrast, we find that in the midbrain-hindbrain boundary, one of the three active enhancers is essential while the other two are dispensable. We further dissect the essential midbrain-hindbrain boundary enhancer to reveal that it is also composed by a mixture of essential and dispensable modules. Cross-species transgenic analysis of this enhancer suggests that its composition may have changed in the vertebrate lineage.

Quality of life in colostomy patients practicing colonic irrigation: An observational study.

AIMS OF THE STUDY: The presence of colostomy has a major impact on quality of life that could potentially be improved by performing colonic irrigation (CI), yet few studies have assessed the impact of this technique on quality of life. The aim of this study was to assess the quality of life between two groups of patients having a colostomy; those practicing CI vs those not practicing CI. PATIENTS AND METHODS: The French Federation of Ostomy (FFO) members were evaluated by a self-questionnaire assessing their experience of CI. Quality of life as assessed by the Stoma-QOL questionnaire was compared between patients practicing CI or not. RESULTS: In total 752 patients were eligible for the study. The median age was 75 years, and 47.26% were men. The median duration between stoma surgery and questionnaire completion was 12.3 years. Forty-one percent of the patients practiced CI. The median quality of life score was significantly higher for the patients practicing the CI: (69.26 vs 58.33, P<0.001). In multivariable analysis, the risk factors for not performing CI were age, obesity, the presence of colostomy for less than six years, and a non-oncologic indication for operation. CONCLUSIONS: CI appeared to improve the quality of life of patients with colostomy. This care is a therapeutic education issue and should be proposed to all patients. Supervision by the enterostomal therapy nurse is recommended especially for patients with a high risk of failure.

Reliability of hemostasis biomarkers is affected by time-dependent intra-patient variability.

Essentials Nucleosomes and free DNA are two newly described biomarkers in venous thromboembolism (VTE). Reliability of nucleosomes, plasma free DNA and conventional hemostasis markers were studied. Hemostasis biological parameters vary over a short time-frame in VTE patients. Hemostasis biological parameters also vary over a short time-frame in healthy controls. SUMMARY: Background Previous studies have associated neutrophil-derived circulating nucleosomes and plasma free DNA with venous thromboembolism (VTE). However, there are few data concerning these two biomarkers and no studies have compared the reliability of nucleosomes and plasma free DNA against that of conventional hemostasis markers. Objectives We performed a 3-year prospective study of nucleosomes and plasma free DNA levels in comparison with conventional hemostatic biomarkers and blood cells. Patients/Methods Fifteen healthy controls and 22 randomly selected patients with a history of VTE were followed monthly for 6 months. The reliability of these markers was evaluated by the intraclass correlation coefficient (ICCs). Results and Conclusions In healthy controls and patients, we found a low reliability for nucleosomes and plasma free DNA, with ICCs at 0.538 (95% confidence interval [CI], 0.334-0.764) and 0.091 (95% CI, -0.026-0.328), respectively, in the healthy controls, and at 0.213 (95% CI, 0.042-0.463) and 0.161 (CI 95%, 0.008-0.398) in the patient group. For the conventional hemostasis biomarkers and for blood cells, reliability ranged from poor to good in the healthy volunteers and from poor to acceptable in the patient group. Our study shows for the first time that hemostasis biological parameters spontaneously vary over a short time-frame in VTE patients and, more surprisingly, in normal individuals. The clinical value of such intra-individual variations is currently unknown. This variability might mean reinterpreting diagnostic or prognostic models based on static evaluation of individuals. Studying the intrinsic value of individual patterns of markers' variability is warranted.

Comparison of two ultrasound guided approaches for axillary vein catheterization: a randomized controlled non-inferiority trial.

BACKGROUND: Axillary vein catheterization via a distal approach is an alternative to the proximal approach to axillary/subclavian vein catheterization under ultrasound (US) guidance. The aim of this trial was to compare the two approaches. METHODS: In a randomized single-centre study, all patients requiring central vein catheterization in intensive care or the operating room were randomly assigned to proximal or distal approach groups. If catheterization failed after two attempts using the approach allocated, the non-allocated approach was used. The primary endpoint was the initial success rate of distal to compared with the proximal approach, using a non-inferiority analysis (lower limit 90% CI greater than -8% non-inferiority margin for group difference). The secondary endpoints were: overall success rates, catheter position and complications. RESULTS: 119/122 included patients were analysed (57 and 62 in the proximal and distal axillary approach groups, respectively). Primary success rates for proximal and distal sites were 87.7 and 85.5%, respectively (difference -2.2%, 90% CI [-12.5-8.1%], non-inferiority P=0.18). The proximal and distal overall success rates were 96.5 and 98.4%, respectively (difference -1.9%, 90% CI [-4.9-8.7%], non-inferiority P<0.01). Thrombogenic catheter positions were 7 (12.3%) in proximal approach group vs 19 (31.7%) in the distal approach group (P=0.01). Complications were comparable in the two groups (2 (3.3%) vs 4 (6.5%), P=0.68). CONCLUSION: In terms of absolute and overall success rates, a distal approach is not non-inferior to a proximal approach. Although associated with a more thrombogenic catheter extremity position, the distal approach can be considered as a rescue alternative after failure of a proximal approach. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT01543360.

Evaluation of in vivo properties of Velosorb™ fast versus Vicryl™ rapide in a functional rabbit model of intradermic suture and episiotomy.

OBJECTIVE: Fast-absorbing polyglactin 910 is commonly used to suture skin and vaginal incisions. In vitro studies have shown similar tensile strength properties between Velosorb™ Fast and Vicryl™ Rapide (Al-Qattan MM. J Hand Surg Br Eur. 2005;30(1):90-91). This randomized study compared Velosorb™ Fast (Covidien) with Vicryl™ Rapide (Ethicon) in a rabbit model of intradermic suture and episiotomy. MATERIALS AND METHODS: This randomized prospective study was performed on 20 New Zealand rabbits, in which we realized a mediolateral perineo-vaginal incision and two vertical skin incisions of about 8 cm on the abdominal wall. The sutures were randomized between Velosorb™ Fast (Covidien) and Vicryl™ Rapide (Ethicon), and each rabbit was treated with the same suture on both vaginal incision and left abdominal incision, while the right abdominal incision was closed with an alternate suture. A macroscopic examination was performed on days 5 and 11. On day 11, animals were sacrificed for histological (Badylak quantitative score) and electron microscopic examinations. Differences were considered significant at p < .05. RESULTS: All of the rabbits survived and were included in the final results. Fifty-one sutures were visible by histologic analysis (27 in the Velosorb™ Fast group and 24 in the Vicryl™ Rapide group). The power of the study was ≥80%. There was no significant difference between the two groups based on the total histologic Badylak score analysis or the Badylak sub-score analysis. CONCLUSIONS: Velosorb™ Fast seems to be as efficient as Vicryl™ Rapide for use in perineal and skin closure.

Bedside tested ocular motor disorders in multiple sclerosis patients.

Background/Aims. Ocular motor disorders (OMDs) are a common feature of multiple sclerosis (MS). In clinical practice, if not reported by patients, OMDs are often underdiagnosed and their prevalence is underestimated. Methods. We studied 163 patients (125 women, 76.7%, 38 men, 23.3%; median age 45.0 years; median disease duration 10 years; median EDSS 3.5) with definite MS (n = 150, 92%) or clinically isolated syndrome (n = 13, 8%) who underwent a thorough clinical examination of eye movements. Data on localization of previous relapses, MS subtype, and MRI findings were collected and analyzed. Results. Overall, 111/163 (68.1%) patients showed at least one abnormality of eye movement. Most frequent OMDs were impaired smooth pursuit (42.3%), saccadic dysmetria (41.7%), unilateral internuclear ophthalmoplegia (14.7%), slowing of saccades (14.7%), skew deviation (13.5%), and gaze evoked nystagmus (13.5%). Patients with OMDs had more severe disability (P = 0.0005) and showed more frequently infratentorial MRI lesions (P = 0.004). Localization of previous relapses was not associated with presence of OMDs. Conclusion. OMDs are frequent in patients with stable (no relapses) MS. A precise bedside examination of eye motility can disclose abnormalities that imply the presence of subclinical MS lesions and may have a substantial impact on definition of the diagnosis and on management of MS patients.

[Functional symptoms and associations of women with genital prolapse]. / Prévalence des troubles fonctionnels et associations anatomo-fonctionnelles chez les femmes présentant un prolapsus génital.

OBJECTIVE: To assess functional symptoms related to genital prolapse and to test anatomo-functional associations. PATIENTS AND METHODS: Observational study, performed between January 2005 and June 2012, on all patients operated for prolapse in a French tertiary referral centre. Data were collected from standardized patients' notes, including baseline characteristics, complete interview on urinary and colo-rectal functional symptoms, MHU score, and POP-Q (Pelvic Organ Prolapse Quantification) clinical evaluation. RESULTS: Three hundred and seventy-four patients, with a mean age of 65.1 years old, mean parity of 2.5, and mean BMI of 25.4, were included. These patients were post-menopausal in 92.5% of cases. Urinary symptoms were: SUI in 30.5%, urgencies in 44.4%, and voiding difficulties in 38.8%. Colo-rectal symptoms, such as defecatory dysfunction and anal incontinence, occurred in 25.1% and 18.5%, respectively. On clinical examination, anterior vaginal wall prolapses were the most common (74.1%). Patients with stage 3-4 cystocele suffered significantly more frequently of nocturia (P=0.04), voiding difficulties (P=0.04), and occult stress urinary incontinence (P<0.001). Patients with stage 3-4 rectocele suffered significantly more frequently of defecatory dysfunction (P=0.005) and performed more often maneuver for defecation (P<0.001). CONCLUSION: Urinary and colo-rectal symptoms are commonly associated with genital prolapse. Anatomo-functional associations were shown regarding different prolapse types and stages. LEVEL OF PROOF: 4.

Preoperative fasting does not affect haemodynamic status: a prospective, non-inferiority, echocardiography study.

BACKGROUND: The link between preoperative fasting and hypovolaemia remains unclear. We tested the hypothesis that preoperative fasting does not significantly increase the proportion of patients with hypovolaemia according to transthoracic echocardiography (TTE) criteria. METHODS: Patients of ASA status I-III and without bowel preparation were included in a non-inferiority, prospective, single-centre trial. Patients underwent passive leg raising (PLR) test and TTE at admission (Day 0) and after 8 h fasting (Day 1). The primary hypothesis was that an 8 h preoperative fasting does not increase the proportion (margin=5%) of patients with a positive PLR test ('functional approach'). The secondary hypothesis was that echocardiographic filling pressures or stroke volume (margin 10%) are not affected by preoperative fasting ('static approach'). RESULTS: One hundred patients were included and 98 analysed. After an 8 h fasting, the change in the proportion of responders to PLR was -6.1% [95% confidence interval (CI)=-16.0 to 3.8] of responders to PLR test on Day 0 when compared with Day 1. Because 95% CI was strictly inferior to 5%, there was no significant increase in the proportion of PLR responders on Day 1 when compared with Day 0. The 95% CI changes of static variables were always fewer than 10%, meaning that preoperative fasting induced significantly no relevant changes in static variables. CONCLUSION: Preoperative fasting did not alter TTE dynamic and static preload indices in ASA I-III adult patients. These results suggest that preoperative fasting does not induce significant hypovolaemia. Clinical trial registration NCT 01258361.

[Surgical site infections in vaginal prolapse surgery]. / Infections du site opératoire en chirurgie du prolapsus par voie vaginale.

INTRODUCTION: Vaginal prolapse surgery is at high risk of surgical site infections (SSI) because it's a "clean-contaminated surgery" and it's frequently associated with implantation of meshes. OBJECTIVES: To evaluate the rate of SSI and associated risk factors in vaginal prolapse surgery with mesh support. METHODS: In a retrospective unicenter study, two groups of patients were operated by vaginal route for a pelvic floor reconstructive surgery with mesh support. Colporraphy was made by classic surgical sutures non-coated (Monosyn(®) 3/0, B-Braun) in the first group, and surgical sutures coated with triclosan in the second group. We collected risk factors of SSIs using the procedure of the CCLIN and analyzed the occurrence of SSIs with a statistical comparative univariate analysis. RESULTS: Study included 78 patients in the first group and 72 in the second group. SSIs total rate was 2.6 % (4 of 150), as part of 3 in the group with surgical sutures non-coated and one in the group with surgical sutures coated with triclosan (P=0.62). CONCLUSION: In our study, SSIs rate in vaginal prolapse surgery was twice higher than classic gynecologic surgery. As the interest of using a surgical suture coated with triclosan to reduce SSI has not been demonstrated statistically, we can't recommend it.

Misconceptions about transmission of AIDS and attitudes toward prevention in the French general public.

A survey of the social perception of AIDS and its prevention was carried out in December 1987 in a representative sample of the adult population of the Paris region (France). A significant part of the general public still holds misconceptions about transmission by casual contact and blood donation. Misbeliefs about modes of transmission clearly encourage individuals' willingness to stigmatize AIDS patients and to support the most coercive measures of prevention (such as quarantine). Advertizing efforts to promote use of condoms, sexual education in schools, and systematic HIV screening for pregnant women are the only measures which create a broad consensus independently of sociocultural differences. Socioeconomic status and even religious or political beliefs strongly influence public opinions toward other preventive measures.

[Social acceptability of screening for HIV seropositivity in pregnant women. An inquiry in the Paris region]. / L'acceptabilité sociale du dépistage de la séropositivité pour VIH chez les femmes enceintes. Une enquête en région parisienne.

Current debate about screening strategies for HIV during pregnancy is not limited to clinical and epidemiologic arguments; ethical and social concerns about the risk of stigmatization against HIV carriers are also taken into account. Therefore, data on public perception of AIDS and attitudes toward preventive measures against the disease can be of interest. In December 1987, a survey was carried out, through personal interview, in a representative sample (900 individuals selected by the quota method) of the 10.5 million inhabitants 18 years of age and over, in the Paris region. 9.8% of the women declared having been tested for HIV, at least once, during 1987, and 20% of these tests were part of prenatal care. Mandatory screening for HIV among pregnant women is supported by 78.6% of the respondents while only 64% support screening during premarital examinations and a minority (37.8%) is in favour of mandatory screening for the whole population. 48.5% of respondents share the misperception that transmission from mother to fetus always happens, and 70% think that women who are HIV carriers should renounce pregnancy: willingness to support mandatory screening for pregnant women is significantly higher among individuals who share these two beliefs.

Modifications of sexual behaviour related to AIDS: a survey in Paris region.

A survey on the effects of AIDS upon sexual behaviour has been carried out, in December 1987, in a representative sample (n = 900) of the population of 18 years of age and over of the Paris region. Among the sexually active population, 13.8% have been tested for HIV at least once in 1987, 10.7% denied some opportunity of sexual intercourse during the last 6 months of 1987 because of fear of AIDS, and 11.8% declared regular utilization of condoms. 53.4% of condom users are new users since the last 6 months, and, 46.6% of all condom users are mainly motivated by fear of AIDS. Multivariate analysis shows that being single, declaring multiple sexual partners and having being tested for HIV are the main predictors for use of condoms, suggesting adoption of a rather coherent preventive behaviour in a significant fraction of the sexually 'at risk' population.