Single, short in-del, and copy number variations detection in monogenic dyslipidemia using a next-generation sequencing strategy.

Optimal molecular diagnosis of primary dyslipidemia is challenging to confirm the diagnosis, test and identify at risk relatives. The aim of this study was to test the application of a single targeted next-generation sequencing (NGS) panel for hypercholesterolemia, hypocholesterolemia, and hypertriglyceridemia molecular diagnosis. NGS workflow based on a custom AmpliSeq panel was designed for sequencing the most prevalent dyslipidemia-causing genes (ANGPTL3, APOA5, APOC2, APOB, GPIHBP1, LDLR, LMF1, LPL, PCSK9) on the Ion PGM Sequencer. One hundred and forty patients without molecular diagnosis were studied. In silico analyses were performed using the NextGENe software and homemade tools for detection of copy number variations (CNV). All mutations were confirmed using appropriate tools. Eighty seven variations and 4 CNV were identified, allowing a molecular diagnosis for 40/116 hypercholesterolemic patients, 5/13 hypocholesterolemic patients, and 2/11, hypertriglyceridemic patients respectively. This workflow allowed the detection of CNV contrary to our previous strategy. Some variations were found in previously unexplored regions providing an added value for genotype-phenotype correlation and familial screening. In conclusion, this new NGS process is an effective mutation detection method and allows better understanding of phenotype. Consequently this assay meets the medical need for individualized diagnosis of dyslipidemia.

Whole-exome sequencing improves the diagnosis yield in sporadic infantile spasm syndrome.

Infantile spasms syndrome (ISs) is characterized by clinical spasms with ictal electrodecrement, usually occurring before the age of 1 year and frequently associated with cognitive impairment. Etiology is widely heterogeneous, the cause remaining elusive in 40% of patients. We searched for de novo mutations in 10 probands with ISs and their parents using whole-exome sequencing (WES). Patients had neither consanguinity nor family history of epilepsy. Common causes of ISs were excluded by brain magnetic resonance imaging (MRI), metabolic screening, array-comparative genomic hybridization (CGH) and testing for mutations in CDKL5, STXBP1, and for ARX duplications. We found a probably pathogenic mutation in four patients. Missense mutations in SCN2A (p.Leu1342Pro) and KCNQ2 (p.Ala306Thr) were found in two patients with no history of epilepsy before the onset of ISs. The p.Asn107Ser missense mutation of ALG13 had been previously reported in four females with ISs. The fourth mutation was an in-frame deletion (p.Phe110del) in NR2F1, a gene whose mutations cause intellectual disability, epilepsy, and optic atrophy. In addition, we found a possibly pathogenic variant in KIF3C that encodes a kinesin expressed during neural development. Our results confirm that WES improves significantly the diagnosis yield in patients with sporadic ISs.

Antibodies to clustered acetylcholine receptor: expanding the phenotype.

BACKGROUND AND PURPOSE: To provide a detailed phenotypical description of seronegative patients with generalized myasthenia gravis and antibodies to clustered acetylcholine receptors (AChRs) and to assess their frequency amongst a French seronegative generalized myasthenia gravis (SNMG) population. METHODS: A French SNMG database was created and the sera from the 37 patients included in it were analysed by immunofluorescence of cell-based assays using cotransfection of AChR subunit genes together with rapsyn to densely cluster the AChRs. RESULTS: Sixteen per cent (n = 6) of the SNMG patients were found to have antibodies to clustered AChR. They presented either with early onset MG and thymic hyperplasia, late onset MG and thymic involution, or thymoma associated MG. They responded well to cholinesterase inhibitors and immunosuppressants. CONCLUSIONS: Patients with antibodies to clustered AChR account for a significant proportion of SNMG patients and resemble patients with AChR antibodies detected by standard radio-immunoprecipitation.

[Outpatient hysterectomy: criteria for acceptability and feasibility, survey among 152 surgeons]. / Critères d'acceptabilité et de faisabilité de l'hystérectomie en ambulatoire, enquête auprès de 152 chirurgiens.

OBJECTIVE: The aim of this study is to determine one-day hysterectomy's criteria of acceptability and feasibility. MATERIALS AND METHODS: We realized an observational descriptive survey based on questionnaires which were sent to gynecologic surgeons. Criteria were defined as major when rate of favorable responses was superior to 70%. RESULTS: Main major criteria were: definition of an age limit (80.3% of respondents), of a Body Mass Index limit (70%), no history of coronary artery disease (77.6%), no anticoagulant therapy with curative intent (95.4%) or antiplatelet (71.1%), no history of sleep apnea (77.4%), surgery within two hours (85.1%), definition of intraoperative bleeding limit (87.5%), no laparotomy (97.4%), no intra abdominal drainage (77.6%), presence of an accompanying at home (99.3%), pain evaluation (97.4%), absence of nausea before leaving (75.5%) and spontaneous urination (96.7%). CONCLUSION: Our study determined major criteria to practice one-day hysterectomy. Decision should be based on a personalized benefice-risk balance analysis. Final decision belongs to patient, as her complete engagement is fundamental.

Antagonistic functions of SET-2/SET1 and HPL/HP1 proteins in C. elegans development.

Cellular identity during metazoan development is maintained by epigenetic modifications of chromatin structure brought about by the activity of specific proteins which mediate histone variant incorporation, histone modifications, and nucleosome remodeling. HP1 proteins directly influence gene expression by modifying chromatin structure. We previously showed that the Caenorhabditis elegans HP1 proteins HPL-1 and HPL-2 are required for several aspects of post-embryonic development. To gain insight into how HPL proteins influence gene expression in a developmental context, we carried out a candidate RNAi screen to identify suppressors of hpl-1 and hpl-2 phenotypes. We identified SET-2, the homologue of yeast and mammalian SET1, as an antagonist of HPL-1 and HPL-2 activity in growth and somatic gonad development. Yeast Set1 and its mammalian counterparts SET1/MLL are H3 lysine 4 (H3K4) histone methyltransferases associated with gene activation as part of large multisubunit complexes. We show that the nematode counterparts of SET1/MLL complex subunits also antagonize HPL function in post-embryonic development. Genetic analysis is consistent with SET1/MLL complex subunits having both shared and unique functions in development. Furthermore, as observed in other species, we find that SET1/MLL complex homologues differentially affect global H3K4 methylation. Our results suggest that HP1 and a SET1/MLL-related complex may play antagonistic roles in the epigenetic regulation of specific developmental programs.

[Management of the HELLP syndrome]. / Prise en charge du HELLP syndrome.

Defined by the association of hemolysis, hepatic dysfunction and thrombocytopenia, the Hemolysis, Elevated Liver enzyme, Low Platelets (HELLP) syndrome can complicate preeclampsia and worsen maternal and fetal prognosis. It can be diagnosed in the immediate postpartum (30%) or in the absence of preeclampsia (10-20%). Clinical diagnosis can be difficult because there is no specific symptom. Abdominal pain or vomiting during the third trimester must lead to think about this diagnosis. Biological criteria are well defined: hemolysis by the presence of schistocytes, increased serum total bilirubin >12 mg/L or LDH >600 IU/L, hepatic dysfunction by increased transaminases and thrombocytopenia by a platelet count <100,000/microL. The evolution of those parameters is a major prognostic factor. With the HELLP syndrome, maternal morbidity is dramatically increased compared to isolated preeclampsia with complications such as eclampsia, placental abruptio, disseminated intravascular coagulation, pulmonary edema, acute renal insufficiency, subcapsular liver hematoma. The management of a HELLP syndrome requests level 3 hospital with intensive care units for neonate and mother. The treatment of this syndrome requires termination of the pregnancy as soon a possible, either by cesarean section or by vaginal delivery if cervical conditions are optimal (without any maternal or fetal complications). Before 32 weeks, a more expectative attitude could be acceptable with the prematurity permitting corticotherapy for fetal pulmonary maturation. This corticotherapy can improve temporary biological parameters but there are no proven benefits to consider improvement for long term maternal or fetal prognosis. During the postpartum, evolution is usually spontaneously favorable. Recurrences are not frequent.

[Magnesium sulphate and severe preeclampsia: its use in current practice]. / Sulfate de magnésium et pré-éclampsie sévère. Innocuité en pratique courante dans des indications ciblées.

OBJECTIVE: To evaluate indications, mode of administration and safety of magnesium sulphate in severe preeclampsia. MATERIAL AND METHODS: We conducted a retrospective descriptive study from January 2000 to December 2002, including patients with severe preeclampsia which was defined as elevated blood pressure >or=140 and/or 90 mmHg with proteinuria >or=0.3g per day, associated with one or more of the following: elevated blood pressure >or=170 and/or 110 mmHg, proteinuria>3g per day, functional symptoms such as headache, hyperreflexia, oliguria<500 ml per day, thrombocytopenia, creatinine level>100 micromol/l, HELLP syndrome. We studied a group of 57 women treated by magnesium sulphate (intravenous bolus of 4.5g during 20 minutes followed by a perfusion of 1.5g/h) associated or not with an antihypertensive treatment. RESULTS: Treatment by magnesium sulphate was started antenatally in 53 women or during immediate postpartum in 4, associated (n=25) or not (n=32) with an antihypertensive treatment. Hyperreflexia was the main indication to start magnesium sulphate treatment (75%). About half (47%) of the cases occurred before 33 weeks of gestation No eclampsia occurred in this group. There was one overdosage which regressed when perfusion was stopped. One patient presented minor side effects attributed to magnesium sulphate. CONCLUSION: Providing a rigorous protocol, indications of magnesium sulfate therapy in severe preeclampsia are well defined. It seems that this treatment could be easily used without severe complications and major side effects.

Bioabsorbable materials for guided bone regeneration prior to implant placement and 7-year follow-up: report of 14 cases.

BACKGROUND: The purpose of the present study was to evaluate the efficacy of a guided bone regeneration (GBR) procedure prior to implant placement and the long-term outcome of the inserted implants. METHODS: Prior to dental implant placement, GBR procedure was performed on 14 patients (mean age 48 years) using a synthetic hydroxyapatite (HA) spacer under a collagen membrane. After a mean healing period of 8 months, bone biopsies were obtained during the placement of 14 implants. The specimens were processed for histology without demineralization in order to assess bone quality and quantity of the regenerated bone. RESULTS: Both the bone density and the resorption degree of HA particles were relatively varied between samples. The different phenotypes of osteoclasts and multinucleated giant cells and the individual host response could partially explain the unpredictable results in terms of bone remodeling and biomaterial resorption. However, the presence of HA particles in the regenerated bone had no influence on the osseointegration of implants presenting a success rate of 86% after a 7-year observation period. CONCLUSIONS: These results confirm the possibility of regenerating bone by means of bioabsorbable materials, assuring at the same time the long-term success for implants inserted in regenerated sites.

A multicenter report on 1,022 consecutively placed ITI implants: a 7-year longitudinal study.

The aim of this multicenter study was to evaluate cumulative success and survival rates of ITI implants after 7 years. A complete medical report was obtained for all 440 patients enrolled in this investigation, which involved 10 different private practices. The 1,022 consecutively placed implants were distributed between completely edentulous, partially edentulous, and single-tooth replacement cases. During the annual follow-up visit, each implant was examined both clinically and radiographically using predefined success criteria. The cumulative survival and success rates were calculated for all implants. Implant subgroups were defined according to the medical history of the patients or pooled according to various indications, locations, implant designs, or implant lengths. In each subgroup, the related cumulative success rate was statistically compared to the global cumulative success rate. Fifteen implants (1.4%) were regarded as early failures, and at the end of the follow-up, the global failure rate reached 6.6%; 30 implants (3%) were lost to follow-up. At 5 years, the cumulative survival rate was 95.4%; this declined to 92.2% at 7 years. The weakest success rates were observed for implants placed in older patients, periodontally treated patients, and completely edentulous arches. Conversely, cumulative success rates that were significantly above average were observed for patients between 40 and 60 years old without pathology, implants placed after bone regeneration, solid-screw implants, implants placed in edentulous spaces, and implants placed as single-tooth replacements. This investigation has demonstrated that in these 10 private practice settings, the success rate for ITI implants remained high for up to 5 years and declined slightly between 5 and 7 years. It should be noted that in later year intervals, a relatively small number of implants remained for the analysis of cumulative success rates.

[Vascular Ehlers-Danlos syndrome and pregnancy: an obstetrical specific support]. / Syndrome d'Ehlers-Danlos de type vasculaire et grossesse : une prise en charge obstétricale spécifique.

The Ehlers-Danlos syndrome (EDS) is a rare inheritable disease, characterised by a defect in collagen synthesis. Various types have been described and the type IV or vascular type is the most severe characterised by vascular, gastrointestinal and gynaecologic complications. We describe in a case report the specific obstetrical support we applied to avoid the most frequent complications such as early spontaneous abortions, pre-term delivery, tearing of perineum, uterine and vascular rupture and hard healing. Pregnancy is very risky in women with vascular EDS. Combination of multidisciplinary support and advice of the rare vascular disease national reference centre may reduce the morbi-mortality rate, including celiprolol long-term treatment.

[Acute pulmonary edema and pregnancy: a descriptive study of 15 cases and review of the literature]. / Œdème aigu du poumon et grossesse : étude descriptive de 15 cas et revue de la littérature.

OBJECTIVE: To describe the incidence and the etiologies of acute pulmonary edema (APE) and the diagnostic procedure used during pregnancy and immediate post-partum. MATERIALS AND METHODS: We analyzed records from a search of codes of heart failure and APE as well as from the term "pulmonary edema" in computerized obstetric records from 2002 to 2010 in a university center of level 3. We identified maternal characteristics, the term of appearance and route of delivery, the time between symptoms and diagnosis, additional tests performed, and data from echocardiography. RESULTS: Fifteen patients had an APE during pregnancy or in the immediate post-partum period during the study period (0.05%). The mean age was 28.6 years and the mean term of appearance was 31.2±3.1 weeks of amenorrhea. The diagnosis was made in 11 cases (73.3%) before delivery and in four during post-partum. The main etiology was preeclampsia (46.6%) followed by heart disease (26.7%), then tocolysis and overfilling (13.3%). In 55% of cases, we found a diagnostic wander characterized by carrying out further unnecessary tests. The echocardiography has led to a change in management in 27.3% of cases. CONCLUSION: The APE is a rare event during pregnancy and the post-partum period and its main etiology is preeclampsia. Some other etiologies are avoidable like the use of beta-agonists by intravenous route. The diagnosis is sometimes difficult, but the realization of a chest X-ray, a simple and inexpensive test, is enough to confirm it.

[French national survey on remifentanil utilisation for obstetrical peridural analgesia]. / Enquête nationale sur l'utilisation du rémifentanil comme alternative à l'analgésie péridurale obstétricale dans les maternités de niveau II et III.

OBJECTIVES: The last French survey on alternatives to neuraxial anaesthesia for labour pain was published in 1997. However, intravenous remifentanil has become increasingly used as an option for labour analgesia. We evaluated the use of remifentanil as an alternative to epidural analgesia in level 2 and 3 French maternities in 2009. STUDY DESIGN: This was an internet-based French survey performed in 2009 including all level 2 and 3 maternities. Data recorded were maternity unit characteristic, alternatives to neuraxial analgesia used, and remifentanil administration protocols. RESULTS: Two hundred and forty maternity units received the survey and 103 responses were completed. A written institutional alternative analgesia protocol for labour pain was present in 78%. Alternative labour analgesia included intermittent nitrous oxide inhalation (58%), intravenous nalbuphine (52%), patient-controlled intravenous administration of remifentanil (52%) and sufentanil (46%). Pethidine administration was reported by one maternity unit (1%). The bolus dose of remifentanil scheduled, and background infusion varied widely between maternity units. The analgesic efficacy of remifentanil used for labour pain was evaluated as moderate (55%) or good (43%). Two serious adverse events were reported. CONCLUSION: Intravenous administration of remifentanil was largely reported as an alternative to neuraxial anaesthesia for labour pain. Although remifentanil administration was most often based on a local written protocol, bolus dose and background infusion varied widely between maternity units.

[Iron deficiency anemia and pregnancy. Prevention and treatment]. / Anémie par carence martiale et grossesse. Prévention et traitement.

OBJECTIVE: To assess the effectiveness and the safety of prevention and treatment of iron deficiency anemia during pregnancy. METHODS: French and English publications were searched using PubMed and Cochrane library. RESULTS: Early screening of iron deficiency by systematic examination and blood analysis seemed essential. Maternal and perinatal complications were correlated to the severity and to the mode of appearance of anemia. Systematic intakes of iron supplements seemed not to be recommended. In case of anemia during pregnancy, iron supplementation was not associated with a significant reduction in substantive maternal and neonatal outcomes. Oral iron supplementation increased blood parameters but exposed to digestive side effects. Women who received parenteral supplementation were more likely to have better hematological response but also severe potential side effects during pregnancy and in post-partum. The maternal tolerance of anemia motivated the choice between parenteral supplementation and blood transfusion. CONCLUSION: Large and methodologically strong trials are necessary to evaluate the effects of iron supplementation on maternal health and pregnancy outcomes.

[Early postpartum management of patients with severe preeclampsia]. / Devenir à court terme des patientes ayant développé une prééclampsie sévère.

The delivery of the foetus and placenta remains the curative treatment for PE, usually allowing a spontaneous, quick and complete regression of all clinical signs and biological anomalies within the first days. However, the risk of developing complications associated with the condition persists mainly during the first 48 to 72 hours and up to one week post partum. Post partum haemodynamic upset and the administered therapeutic measures are predisposing factors to these complications. This critical period therefore requires intensive monitoring and the delivery of appropriate treatments.