Accuracy profile validation of a new method for carbon monoxide measurement in the human blood using headspace-gas chromatography-mass spectrometry (HS-GC-MS).

The aim of our study was to provide an innovative headspace-gas chromatography-mass spectrometry (HS-GC-MS) method applicable for the routine determination of blood CO concentration in forensic toxicology laboratories. The main drawback of the GC/MS methods discussed in literature for CO measurement is the absence of a specific CO internal standard necessary for performing quantification. Even if stable isotope of CO is commercially available in the gaseous state, it is essential to develop a safer method to limit the manipulation of gaseous CO and to precisely control the injected amount of CO for spiking and calibration. To avoid the manipulation of a stable isotope-labeled gas, we have chosen to generate in a vial in situ, an internal labeled standard gas ((13)CO) formed by the reaction of labeled formic acid formic acid (H(13)COOH) with sulfuric acid. As sulfuric acid can also be employed to liberate the CO reagent from whole blood, the procedure allows for the liberation of CO simultaneously with the generation of (13)CO. This method allows for precise measurement of blood CO concentrations from a small amount of blood (10 μL). Finally, this method was applied to measure the CO concentration of intoxicated human blood samples from autopsies.

Ossification du ligament commun cervical postérieur : coexistence de spondylarthrite et d'une hyperostose idiopathique diffuse du squelette

Introduction: L'ossification du ligament commun vertébral postérieur (LCVP) est une hyperostose prédominant au rachis cervical, associée à différentes pathologies constructrices comme l'arthrose, la maladie de Forestier (ou DISH : Diffuse Idiopathic Squeletal Hyperostosis) ou les spondylarthrites (1). Nous rapportons le cas d'une patiente avec ossification cervicale du LCVP, avec coexistence de DISH et de spondylarthrite.Observation: Patiente de 55 ans, d'origine Irakienne, qui présente depuis l'âge de 40 ans des lombalgies et des talalgies attribuées initialement à un métier physique. En 2008, apparait une cervicobrachialgie C6 gauche. L'IRM cervicale retrouve plusieurs hernies discales (C5-C6 et C6-C7 gauches) avec un canal cervical étroit constitutionnel et une ossification du LCVP cervical. Cette ossification est attribuée à un DISH (Figure1). L'échec du traitement conservateur de la névralgie et la menace neurologique de l'ossification du LCVP conduisent en 2010 à une laminectomie C3-C6 avec fixation postérieure (figure 2). En 2011, la patiente consulte en raison de la persistance de cette névralgie, ainsi que pour des lombalgies inflammatoires. La coexistence de ces symptômes inflammatoires et de l'ossification du LCVP nous incite à réaliser une IRM des sacroiliaques qui retrouve une sacroiliite bilatérale (figure 3). Le diagnostic de spondylarthrite HLA B27 négative est retenu devant l'association des signes cliniques et radiologiques. L'étiologie précise quant à l'origine de l'ossification du LCVP reste incertaine, néanmoins un traitement spécifique de la spondylarthopathie est proposé.Discussion: L'ossification du ligament vertébral commun postérieur est une hyperostose dont les principales causes sont le DISH et les spondylarthrites. On retrouve une ossification du ligament commun vertébral, qu'il soit lombaire, cervical ou dorsal, dans 10 à 50 % des DISH, et 3.5 à 30% des spondylarthrites. 4 types d'ossification du LCVP sont décrites : continue, segmentaire, mixte ou circonscrite. Selon Resnick, le DISH se différencie de la spondylarthrite par la présence d'ossifications antérolatérales d'au moins 4 vertèbres contigües sans érosion des sacroiliaques. Cependant on retrouve dans la littérature quelques cas décrivant la coexistence des 2 pathologies. Récemment, Kim et al. Ont rapporté un cas similaire à notre patiente, avec ossification du LCVP cervical et coexistence de DISH et de spondylarthrite (2).Conclusion: Devant la présence d'une ossification du ligament vertébral commun postérieur cervical, il convient de rechercher des signes de DISH et de spondylarthrite, car leur coexistence est possible. La prévalence exacte de cette association reste encore à déterminer.

A method to obtain the integrated contour image in colour scenes

This paper describes a method to achieve the most relevant contours of an image. The presented method proposes to integrate the information of the local contours from chromatic components such as H, S and I, taking into account the criteria of coherence of the local contour orientation values obtained from each of these components. The process is based on parametrizing pixel by pixel the local contours (magnitude and orientation values) from the H, S and I images. This process is carried out individually for each chromatic component. If the criterion of dispersion of the obtained orientation values is high, this chromatic component will lose relevance. A final processing integrates the extracted contours of the three chromatic components, generating the so-called integrated contours image

Uptake of macromolecules by cercariae during skin penetration and transformation to schistosomula (Schistosoma mansoni)

Here, we observed the uptake of membrane-impermeant molecules by cercariae as they penetrate the skin and are transformed into schistosomula. We propose that membrane-impermeant molecules, Lucifer Yellow, Propidium iodide and Hoechst 33258 enter the parasite through both thenephridiopore and the surface membrane and then diffuse throughout the body of the parasite. We present a hypothesis that the internal cells of the body of the schistosomulum represent a new host-parasite interface, at which skin-derived growth factors may stimulate receptors on internal membranes during transformation of the cercariae into the schistosomulum.

An improved method for discovering link criticality in transport networks

Quantitatively assessing the importance or criticality of each link in a network is of practical value to operators, as that can help them to increase the network's resilience, provide more efficient services, or improve some other aspect of the service. Betweenness is a graph-theoretical measure of centrality that can be applied to communication networks to evaluate link importance. However, as we illustrate in this paper, the basic definition of betweenness centrality produces inaccurate estimations as it does not take into account some aspects relevant to networking, such as the heterogeneity in link capacity or the difference between node-pairs in their contribution to the total traffic. A new algorithm for discovering link centrality in transport networks is proposed in this paper. It requires only static or semi-static network and topology attributes, and yet produces estimations of good accuracy, as verified through extensive simulations. Its potential value is demonstrated by an example application. In the example, the simple shortest-path routing algorithm is improved in such a way that it outperforms other more advanced algorithms in terms of blocking ratio

Estimation of the probability of congestion using Monte Carlo method in OPS networks

In networks with small buffers, such as optical packet switching based networks, the convolution approach is presented as one of the most accurate method used for the connection admission control. Admission control and resource management have been addressed in other works oriented to bursty traffic and ATM. This paper focuses on heterogeneous traffic in OPS based networks. Using heterogeneous traffic and bufferless networks the enhanced convolution approach is a good solution. However, both methods (CA and ECA) present a high computational cost for high number of connections. Two new mechanisms (UMCA and ISCA) based on Monte Carlo method are proposed to overcome this drawback. Simulation results show that our proposals achieve lower computational cost compared to enhanced convolution approach with an small stochastic error in the probability estimation

Granular layer in the periplasmic space of gram-positive bacteria and fine structures of Enterococcus gallinarum and Streptococcus gordonii septa revealed by cryo-electron microscopy of vitreous sections.

High-resolution structural information on optimally preserved bacterial cells can be obtained with cryo-electron microscopy of vitreous sections. With the help of this technique, the existence of a periplasmic space between the plasma membrane and the thick peptidoglycan layer of the gram-positive bacteria Bacillus subtilis and Staphylococcus aureus was recently shown. This raises questions about the mode of polymerization of peptidoglycan. In the present study, we report the structure of the cell envelope of three gram-positive bacteria (B. subtilis, Streptococcus gordonii, and Enterococcus gallinarum). In the three cases, a previously undescribed granular layer adjacent to the plasma membrane is found in the periplasmic space. In order to better understand how nascent peptidoglycan is incorporated into the mature peptidoglycan, we investigated cellular regions known to represent the sites of cell wall production. Each of these sites possesses a specific structure. We propose a hypothetic model of peptidoglycan polymerization that accommodates these differences: peptidoglycan precursors could be exported from the cytoplasm to the periplasmic space, where they could diffuse until they would interact with the interface between the granular layer and the thick peptidoglycan layer. They could then polymerize with mature peptidoglycan. We report cytoplasmic structures at the E. gallinarum septum that could be interpreted as cytoskeletal elements driving cell division (FtsZ ring). Although immunoelectron microscopy and fluorescence microscopy studies have demonstrated the septal and cytoplasmic localization of FtsZ, direct visualization of in situ FtsZ filaments has not been obtained in any electron microscopy study of fixed and dehydrated bacteria.

Rotavirus A genotype G1P[8]: a novel method to distinguish wild-type strains from the Rotarix® vaccine strain

Rotaviruses are important enteric pathogens for humans and animals. Group A rotaviruses (RV-A) are the most common agents of severe gastroenteritis in infants and young children and vaccination is the most effective method to reduce RV-A-associated diseases. G1P[8], the most prevalent RV-A genotype worldwide, is included in the RV-A vaccine Rotarix®. The discrimination between wild-type G1P[8] and vaccine G1P[8] strains is an important topic in the study of RV-A epidemiology to manage outbreaks and to define control measures for vaccinated children. In this study, we developed a novel method to segregate the wild-type and vaccine strains using restriction endonucleases. The dsRNA from the Rotarix® vaccine was sequenced and the NSP3 gene was selected as the target gene. The vaccine strain has a restriction pattern that is different than that of wild-type RV-A G1P[8] isolates after digestion with the restriction endonuclease BspHI. This pattern could be used as a marker for the differentiation of wild-type G1P[8] strains from the vaccine strain.

A New Method for ECG Tracking of Persistent Atrial Fibrillation Termination during Stepwise Ablation

Stepwise radiofrequency catheter ablation (step-CA) has become the treatment of choice for the restoration of sinus rhythm (SR) in patients with long-standing persistent atrial fibrillation (pers-AF). Its success rate appears limited as the amount of ablation to achieve long term SR is unknown. Multiple organization indexes (OIs) have been previously developed to track the organization of AF during step-CA, however, with limited success. We report an adaptive method for tracking AF termination (AF-term) based on OIs characterizing the relationship between harmonic components of atrial activity from the surface ECG of AF activity. By computing their relative evolution during the last two steps preceding AF-term, we found that the performance of our OIs was superior to classical indices to track the efficiency of step-CA "en route" to AF-term. Our preliminary results suggest that the gradual synchronization between the fundamental and its first harmonic of AF activity appears as a promising parameter for predicting AF-term during step-CA.

Schistosoma mansoni: a method for inducing resistance to praziquantel using infected Biomphalaria glabrata snails

To elucidate the mechanisms of antischistosoma resistance, drug-resistant Schistosoma mansoni laboratory isolates are essential. We developed a new method for inducing resistance to praziquantel (PZQ) using successive drug treatments of Biomphalaria glabrata snails infected with S. mansoni. Infected B. glabrata were treated three times with 100 mg/kg PZQ for five consecutive days with a one-week interval between them. After the treatment, the cercariae (LE-PZQ) produced from these snails and the LE strains (susceptible) were used to infect mice. Forty-five days after infection, mice were treated with 200, 400 or 800 mg/kg PZQ. Thirty days post-treatment, we observed that the mean number of worms recovered by perfusion was significantly higher in the group of mice infected with the LE-PZQ isolate treated with 200 and 400 mg/kg in comparison to the LE strain with the same treatment. Moreover, there was a significant difference between the ED50 (effective dose required to kill 50% of the worms) of the LE-PZQ isolate (362 mg/kg) and the LE strain (68 mg/kg). In the in vitro assays, the worms of the LE-PZQ isolate were also less susceptible to PZQ. Thus, the use of infected snails as an experimental model for development of resistance to S. mansoni is effective, fast, simple and cheap.

Use of accuracy profile for the validation of a gas chromatography-negative chemical ionization tandem mass spectrometry method: quantification of ethyl glucuronide in hair

Introduction: Ethylglucuronide (EtG) is a direct and specific metabolite of ethanol. Its determination in hair is of increasing interest for detecting and monitoring alcohol abuse. The quantification of EtG in hair requires analytical methods showing highest sensitivity and specificity. We present a fully validated method based on gas chromatography-negative chemical ionization tandem mass spectrometry (GC-NCI-MS/MS). The method was validated using French Society of Pharmaceutical Sciences and Techniques (SFSTP) guidelines which are based on the determination of the total measurement error and accuracy profiles. Methods: Washed and powdered hair is extracted in water using an ultrasonic incubation. After purification by Oasis MAX solid phase extraction, the derivatized EtG is detected and quantified by GC-NCI-MS/MS method in the selected reaction monitoring mode. The transitions m/z 347 / 163 and m/z 347 / 119 were used for the quantification and identification of EtG. Four quality controls (QC) prepared with hair samples taken post mortem from 2 subjects with a known history of alcoholism were used. A proficiency test with 7 participating laboratories was first run to validate the EtG concentration of each QC sample. Considering the results of this test, these samples were then used as internal controls for validation of the method. Results: The mean EtG concentrations measured in the 4 QC were 259.4, 130.4, 40.8, and 8.4 pg/mg hair. Method validation has shown linearity between 8.4 and 259.4 pg/mg hair (r2 > 0.999). The lower limit of quantification was set up at 8.4 pg/mg. Repeatability and intermediate precision were found less than 13.2% for all concentrations tested. Conclusion: The method proved to be suitable for routine analysis of EtG in hair. GC-NCI-MS/MS method was then successfully applied to the analysis of EtG in hair samples collected from different alcohol consumers.