Disruption of pathways regulated by Integrator complex in Galloway-Mowat syndrome due to WDR73 mutations.

Several studies have reported WDR73 mutations to be causative of Galloway-Mowat syndrome, a rare disorder characterised by the association of neurological defects and renal-glomerular disease. In this study, we demonstrate interaction of WDR73 with the INTS9 and INTS11 components of Integrator, a large multiprotein complex with various roles in RNA metabolism and transcriptional control. We implicate WDR73 in two Integrator-regulated cellular pathways; namely, the processing of uridylate-rich small nuclear RNAs (UsnRNA), and mediating the transcriptional response to epidermal growth factor stimulation. We also show that WDR73 suppression leads to altered expression of genes encoding cell cycle regulatory proteins. Altogether, our results suggest that a range of cellular pathways are perturbed by WDR73 loss-of-function, and support the consensus that proper regulation of UsnRNA maturation, transcription initiation and cell cycle control are all critical in maintaining the health of post-mitotic cells such as glomerular podocytes and neurons, and preventing degenerative disease.

Cardiac output monitoring with thermodilution pulse-contour analysis vs. non-invasive pulse-contour analysis.

Intravenous fluid boluses guided by changes in stroke volume improve some outcomes after major surgery, but invasive measurments may limit use. From October 2016 to May 2018, we compared the agreement and trending ability of a photoplethysmographic device (Clearsight) with a PiCCO, calibrated by thermodilution, for haemodynamic variables in 20 adults undergoing major elective surgery. We analysed 4519 measurement pairs, including before and after 68 boluses of 250 ml crystalloid. The bias and precision of stroke volume measurement by Clearsight were -0.89 ± 4.78 ml compared with the invasive pulse-contour cardiac output device. The coefficient of agreement for stroke volume variation after fluid boluses between the two devices was 0.79 ('strong'). Fluid boluses that increased stroke volume by ≥ 10% increased mean absolute volume (SD) and mean percentage (SD) stroke volume measurements similarly for the invasive pulse-contour cardiac output and Clearsight devices: 9 (4) ml vs. 8 (4) ml and 16% (8%) vs. 15% (10%), respectively, p > 0.05. The non-invasive Clearsight pulse-contour analysis was similar to an invasive pulse-contour device in measuring absolute and changing stroke volumes during major surgery.

Antibioprophylaxis in surgery and interventional medicine (adult patients). Update 2017.

Infection is a risk for any intervention. In surgery, for example, pathogenic bacteria are found in more than 90% of operative wounds during closure. This exists whatever the surgical technique and whatever the environment (the laminar flow does not entirely eliminate this risk). These bacteria are few in number but can proliferate. They find in the operative wound a favourable environment (haematoma, ischaemia, modification of oxido-reduction potential...) and the intervention induces anomalies of the immune defences. In the case of the installation of foreign material, the risk is increased. The objective of antibiotic prophylaxis (ABP) is to prevent bacterial growth in order to reduce the risk of infection at the site of the intervention. The preoperative consultation represents a privileged moment to decide on the prescription of a ABP. It is possible to define the type of intervention planned, the associated risk of infection (and therefore the necessity or not of ABP), the time of prescription before surgery and any allergic antecedents which may modify the choice of the selected antibiotic molecule.

Continuous monitoring of haemoglobin concentration after in-vivo adjustment in patients undergoing surgery with blood loss.

Non-invasive monitoring of haemoglobin concentration provides real-time measurement of haemoglobin concentration (SpHb) using multi-wavelength pulse co-oximetry. We hypothesised that in-vivo adjustment using the mean of three haemoglobinometer (HemoCue®) measurements from an arterial blood sample at the first SpHb measurement (HCueART) would increase the accuracy of the monitor. The study included 41 adults for a total of 173 measurements of haemoglobin concentration. In-vivo adjusted SpHb was automatically calculated by the following formula: in-vivo adjusted SpHb = unadjusted SpHb - (SpHb - HCueART). The accuracy of in-vivo adjusted SpHb was compared with SpHb retrospectively adjusted using the same formula, except for haemoglobin level which was assessed at the central laboratory and then compared with all other available invasive methods of haemoglobin measurement (co-oximetry, HbSAT; arterial HemoCue, HCueART; capillary HemoCue, HCueCAP). Compared with laboratory measurement of haemoglobin concentration, bias (precision) for unadjusted SpHb, in-vivo adjusted SpHb, retrospectively adjusted SpHb, HbSAT, HCueART and HCueCAP were -0.4 (1.4), -0.3 (1.1), -0.3 (1.1), -0.6 (0.7), 0.0 (0.4) and -0.5 (1.2) g.dl(-1) , respectively. In-vivo adjustment of SpHb values using the mean of three arterial HemoCue measurements improved the accuracy of the device similar to those observed after a retrospective adjustment using central laboratory haemoglobin level.

Colistin in critically ill patients.

Colistin is a re-emerging old antibiotic that is used as a salvage treatment against multidrug-resistant Gram-negative infections. Because it is administrated as an inactive prodrug, colistin methanesulfonate (CMS) that undergoes rapid hydrolyze to colistin, pharmacokinetic studies using biological assays are unreliable. With the recent development of new assays using high performance liquid chromatography (HPLC) accurate pharmacokinetic of CMS and formed colistin is now available in various populations. This article aims to update previous reports on pharmacodynamics, pharmacokinetics, safety and clinical use of colistin, with a special focus on data useful to treat critically ill patients.

An inventory of trace elements inputs to French agricultural soils.

The inputs of ten trace elements (As, Cd, Cu, Cr, Hg, Mo, Ni, Pb, Se, Zn) to French agricultural soils have been assessed. The six main sources considered were: pesticides, mineral fertilizers, animal manure, liming materials, sludge and composts and atmospheric deposition. Data were collected to compute inputs at both national and regional (departmental) scales. The inventory methodology is based on two principles: data are traceable and easy to update. At a national scale, the inventory showed that trace elements inputs can be ranked: Zn≫Cu≫Cr>Pb>Ni>As=Mo>Se>Cd>Hg. Animal manure, mineral fertilizers and pesticides are the predominant sources of TEs. These results are globally in agreement with literature data though atmospheric deposition is shown to be lower than in more industrial countries such as China and United Kingdom where similar surveys were conducted. The inputs of trace elements vary strongly between regions in relation with agricultural activities. This inventory (and the related database) provides basis for developing and monitoring policies to control and reduce trace elements contamination of agricultural soils at both national and regional (departmental) scales.

[Guidelines for point-of-care testing quality management according to ISO 22870]. / Recommandations pour la mise en place d'un système de management de la qualité des examens de biologie médicale délocalisés.

In this paper, we focus on the additional requirements of EN ISO 22870 compared to those described in Chapter 4: Quality Management of EN ISO 15189. They concern the quality policy, the management reviews and the audits. Thus, we propose a template of quality policy statement, and specific requirements for conducting management review of POCT are given. Finally, a questionnaire for performing an audit of POCT activities is proposed. The composition and activities of the multidisciplinary group for the supervision of POCT activities, which is also a specific requirement of EN ISO 22870, is discussed in another article of this volume.

[Practice assessment of central venous lines care in surgical ICU of French university hospitals]. / Evaluation des pratiques de gestion des cathéters veineux centraux dans les réanimations chirurgicales universitaires françaises.

INTRODUCTION: Recommendations on insertion and maintenance of central venous catheters (CVC) in intensive care unit (ICU) patients were updated in 2002. The aim of this study was to estimate their knowledge and/or application by physicians in French university hospital ICUs. METHODS: Two forms were sent to 124 professors of anaesthesia and intensive care encouraging them to participate to the survey. The first one was completed by the physician in charge of each unit and concerned the structure and activity of the unit in 2006. The second one was filled by each junior or senior physician working in the units and asked for experience, CVC insertion modalities and knowledge of CVC care protocols. RESULTS: Forty-one (75 %) university hospitals with at least one adult surgical ICU took part to the study. A questionnaire was filled by 124 senior (75 % of the staff) and 53 junior (43 % of the staff) physicians inserting an average of 10 CVC per month (range, 1-35). A written protocol for CVC insertion was known by 127 (72 %) of them. CVC insertion was done while wearing sterile gown (97 %), cap (100 %) and surgical mask (100 %) and using large sterile drapes (96 %). The antiseptic solution used for cutaneous antisepsis was povidone iodine in aqueous (36 %) or alcoholic solution (40 %), or an alcoholic solution of chlorhexidine (24 %) applied one (9 %), two (64 %) or three (27 %) times before insertion. A 4-times disinfection sequence (washing, rinsing, drying and disinfection) was performed by 161 (91 %) physicians. Ultrasound-guided insertion was realized by only eight (5 %) operators. CVCs were made of polyurethane (84 %), usually multi-lumens (>96 %) and rarely tunnelised (14 %). Only two physicians (1 %) sometimes use catheters coated with antibiotics or antiseptics. The site for catheter insertion was mostly the sub-clavian (47 %) or internal jugular vein (34 %), and rarely the femoral vein (20 %). CVCs were secured with a thread (99 %) and covered with a semi-permeable dressing (76 %). Concerning CVCs maintenance, 91 % of physicians acknowledged the existence of a written protocol in the unit. Dressings were changed every day (10 %), every two days (49 %), every three days (29 %) or every four days or more (12 %) by using the same antiseptic solution and semi-permeable transparent dressing in 78 % of cases. Venous lines changes were done during dressing maintenance (48 %), every day in case of administration of lipids (32 %) or just after administration of blood products via the catheter (32 %). Routine change of CVC was rarely recommended (11 %). CONCLUSION: The high number of answers allows setting of a precise state of CVCs insertion practices in adult surgical ICUs. Recommendations for central venous catheter insertion and maintenance are not still known and\or applied.

Partial sequences of nitrogen metabolism genes in hexaploid wheat.

Our objective was to partially sequence genes controlling nitrogen metabolism in wheat species in order to find sequence polymorphism that would enable their mapping. Primers were designed for nitrate reductase, nitrite reductase, glutamate dehydrogenase and glutamate synthase (GOGAT), and gene fragments were amplified on Triticum aestivum, T. durum, T. monococcum, T. speltoides and T. tauschii. We obtained more than 8 kb of gene sequences, mainly as coding regions (60%). Polymorphism was quantified by comparing two-by-two the three genomes of the hexaploid cultivar Arche and genomes of diploid wheat species. On average, the polymorphism rate was higher for non-coding regions, where it ranged from 1/60 to 1/23, than for coding regions (range: 1/110-1/40) except when the hexaploid D genome was compared to that of T. tauschii (1/800 and 1/816, respectively). Genome-specific primers were devised for the ferredoxin-dependent (Fd)-GOGAT gene, and they enabled the mapping of this gene on homoeologous chromosomes of group 2 using Chinese Spring deletion lines. A single nucleotide polymorphism (SNP) detected between the two hexaploid wheat cultivars Arche and Recital was used to genetically map Fd-GOGAT on chromosome 2D using a population of dihaploid lines. Fd-GOGAT-specific primers were used to estimate the SNP rate on a set of 11 hexaploid and nine Durum wheat genotypes leading to the estimate of 1 SNP/515 bp. We demonstrate that polymorphism detection enables heterologous, homeologous and even paralogous copies to be assigned, even if the elaboration of specific primer pairs is time-consuming and expensive because of the sequencing.

[The interest of cholesterol levels in young children. Study in a population of 4,697 children aged 4]. / Valeur de la cholestérolémie chez l'enfant. Etude sur une population de 4697 enfants de 4 ans.

INTRODUCTION: There is a need to dispose of normal cholesterol levels in young children. In view of the paucity of such data in France, we analysed the results of screening conducted in children. METHOD: We analysed the cholesterol levels of 4697 children, with a mean age of 4.3 years, attending a medical check-up in a Child Health Unit in a National Health Scheme centre in Paris. All the children were recruited consecutively during the year 2002. RESULTS: The mean cholesterol level was of 4.4 mmol/L +/- 0.75. We detected a slight gender-related variation (the mean in girls and boys were of 4.5 +/- 0.76 and 4.4 +/- 0.74 respectively). The 95 percentile in girls and boys were 5.7 and 5.6 mmol/L, respectively. DISCUSSION: For the first time in France, we now have access to data on normal cholesterol levels in a large cohort of 4 year-old children. Screening for hypercholesterolaemia in children provides the opportunity to discuss dietary counselling.

Arabidopsis glucosidase I mutants reveal a critical role of N-glycan trimming in seed development.

Glycoproteins with asparagine-linked (N-linked) glycans occur in all eukaryotic cells. The function of their glycan moieties is one of the central problems in contemporary cell biology. N-glycosylation may modify physicochemical and biological protein properties such as conformation, degradation, intracellular sorting or secretion. We have isolated and characterized two allelic Arabidopsis mutants, gcs1-1 and gcs1-2, which produce abnormal shrunken seeds, blocked at the heart stage of development. The mutant seeds accumulate a low level of storage proteins, have no typical protein bodies, display abnormal cell enlargement and show occasional cell wall disruptions. The mutated gene has been cloned by T-DNA tagging. It codes for a protein homologous to animal and yeast alpha-glucosidase I, an enzyme that controls the first committed step for N-glycan trimming. Biochemical analyses have confirmed that trimming of the alpha1,2- linked glucosyl residue constitutive of the N-glycan precursor is blocked in this mutant. These results demonstrate the importance of N-glycan trimming for the accumulation of seed storage proteins, the formation of protein bodies, cell differentiation and embryo development.

The Arabidopsis AtEPR1 extensin-like gene is specifically expressed in endosperm during seed germination.

Screening of 10 000 Arabidopsis transgenic lines carrying a gene-trap (GUS) construct has been undertaken to identify markers of seed germination. One of these lines showed GUS activity restricted to the endosperm, at the micropylar end of the germinating seed. The genomic DNA flanking the T-DNA insert was cloned by walking PCR and the insertion was shown to be located 70 bp upstream of a 2285 bp open reading frame (AtEPR1) sharing strong similarities with extensins. The AtEPR1 open reading frame consists of 40 proline-rich repeats and is expressed in both wild-type and mutant lines. The expression of the AtEPR1 gene appears to be under positive control of gibberellic acid, but is not downregulated by abscisic acid during seed germination. No expression was detected in organs other than endosperm during seed germination. The putative role of AtEPR1 is discussed in the light of its specific expression in relation to seed germination.

Electroshock afterdischarges are related to neuronal oscillability: neuronal models of Aplysia.

Identifiable giant neurons of Aplysia explored intracellularly behave differently at the offset of an intracellular electroshock (IES) or after a synaptic 'tetanization', according to their functional type: neurons of the stable type depolarize and fire at the offset of the IES, anodal or cathodal, thus eliciting an afterdischarge (AD). The threshold of this AD is lowered if the neuron is destabilized, i.e. converted from the stable to an oscillatory type (for instance by decalcification). Neurons normally of the tonic type are more sensitive to an IES, eliciting a longer afterdischarge than the stable neurons. Extracellular electroshock (EES) anodal or cathodal, applied directly on desheathed somata of Helix give long-lasting afterdischarges at the offset. In addition, EES stimulating presynaptic terminals or axons leads to a high frequency synaptic input on remote neurons. At the offset of this input either prolonged synaptic afterdischarges or postsynaptic rebounds of the membrane potential sustaining bursts of decreasing amplitude denote apparent oscillatory properties of the synaptically activated neuron. Finally, any conversion by convulsants of tonic neurons to oscillators highly facilitates the elicitation of afterdischarges of axons simultaneous to paroxysmal depolarization shifts of the homologous somata. These results indicate that afterdischarge elicitation is highly facilitated (low threshold) in normal oscillatory neurons and/or chemically destabilized neurons.

Collagen synthesis in rat gingiva during tooth movement.

The response of the gingiva to an increased interdental space was studied by creating a diastema between the central incisors of rats and analyzing autoradiographically the incorporation of H3 proline in the gingiva to detect increased collagen production. In addition, conventional histologic methods were used to determine changes in the gingival architecture. The results indicate that the gingiva responds to an increased space in at least two ways. One is the production of more collagen fibers. The other involves the reorientation of the existing fibers in a horizontal plane as the gingival papilla becomes flattened.

Malaria and anemia of pregnancy in an African savanna zone. Epidemiological, hematological, biological and immunological study of 2 villages of the Bamako region, Republic of Mali. / Paludisme et anémie de la grossesse en zone de savane africaine. Etude épidémiologique, hématologique, biologique et immunologique dans deux villages de la région de Bamako, République du Mali.

An haematological, biological, parasitological and immunological study about anaemia of pregnancy was carried out in two rural village of Mali Republic, where P. flaciparum malaria is hyper-endemic. The 25 pregnant women found in the villages were compared with 23 controls. One could observe that anaemia more often normochromic and regenerative usually appears during the second trimestre of pregnancy. Only a few cases with haemaglobin levels below 8 g. % are hypochromic. Serum transferrin levels were slightly higher among pregnant. None among 31 bone marrows examined showed megaloblastic changes. Haptoglobin levels below 10 mg. % were observed in 3/4 of the pregnant women versus 1/4 in controls. Like some others, this study confirms the primary responsability of malaria haemolysis in the aetiology of anaemia of pregnancy and the interest of systematic chemoprophylaxis, at least from the third month of pregnancy. Vitamin and iron therapy is to consider therafter in hyper-endemic areas of P. falciparum malaria when nutritional problems are not predominant.