Comparison of biomarker for diagnosis of nitrous oxide abuse: challenge of cobalamin metabolic parameters, a retrospective study.

BACKGROUND: Recreational use of nitrous oxide (N2O) leads to neurological disorders including combined subacute degeneration of spinal cord, psychological disorders, and thrombosis. Serum or urine N2O assays could not be routinely performed. Hence, it is necessary to investigate other biological markers such as metabolic markers. We aimed here to challenge the three main biological markers used for the diagnosis of nitrous oxide abuse as total vitamin B12, homocysteine, and methylmalonic acid. METHODS: We retrospectively collected clinical and biological data from 52 patients with known, documented chronic N2O abuse and associated clinical signs (peripheral neuropathy disability score or thrombosis event). Sera and plasma total vitamin B12, methylmalonic acid (MMA), and homocysteine were performed to identify the most specific marker of chronic N2O intoxication and related clinical outcomes. RESULTS: Plasma homocysteine was almost consistently increased in case of N2O chronic consumption, whereas MMA increase and total vitamin B12 decrease are not systematically found. Our results showed that none of the markers are correlated with levels of N2O consumptions. However, homocysteine and MMA are correlated with clinical severity, but MMA seems to be a better marker of clinical severity. CONCLUSION: There is no specific marker of nitrous oxide abuse according to levels of consumption, total vitamin B12 decrease could not be used either as consumption or as severity marker. However, we showed that homocysteine is consistently increased and could be used as marker of recent N2O consumption. On the other hand, we showed that MMA could be used as a marker of clinical gravity.

Oligoclonal Band Straightening Based on Optimized Hierarchical Warping for Multiple Sclerosis Diagnosis.

The detection of immunoglobulin G (IgG) oligoclonal bands (OCB) in cerebrospinal fluid (CSF) by isoelectric focusing (IEF) is a valuable tool for the diagnosis of multiple sclerosis. Over the last decade, the results of our clinical research have suggested that tears are a non-invasive alternative to CSF. However, since tear samples have a lower IgG concentration than CSF, a sensitive OCB detection is therefore required. We are developing the first automatic tool for IEF analysis, with a view to speeding up the current visual inspection method, removing user variability, reducing misinterpretation, and facilitating OCB quantification and follow-up studies. The removal of band distortion is a key image enhancement step in increasing the reliability of automatic OCB detection. Here, we describe a novel, fully automatic band-straightening algorithm. The algorithm is based on a correlation directional warping function, estimated using an energy minimization procedure. The approach was optimized via an innovative coupling of a hierarchy of image resolutions to a hierarchy of transformation, in which band misalignment is corrected at successively finer scales. The algorithm's performance was assessed in terms of the bands' standard deviation before and after straightening, using a synthetic dataset and a set of 200 lanes of CSF, tear, serum and control samples on which experts had manually delineated the bands. The number of distorted bands was divided by almost 16 for the synthetic lanes and by 7 for the test dataset of real lanes. This method can be applied effectively to different sample types. It can realign minimal contrast bands and is robust for non-uniform deformations.

Three-dimensional (3D) culture of adult murine colon as an in vitro model of cryptosporidiosis: Proof of concept.

Cryptosporidium parvum is a major cause of diarrheal illness and was recently potentially associated with digestive carcinogenesis. Despite its impact on human health, Cryptosporidium pathogenesis remains poorly known, mainly due to the lack of a long-term culture method for this parasite. Thus, the aim of the present study was to develop a three-dimensional (3D) culture model from adult murine colon allowing biological investigations of the host-parasite interactions in an in vivo-like environment and, in particular, the development of parasite-induced neoplasia. Colonic explants were cultured and preserved ex vivo for 35 days and co-culturing was performed with C. parvum. Strikingly, the resulting system allowed the reproduction of neoplastic lesions in vitro at 27 days post-infection (PI), providing new evidence of the role of the parasite in the induction of carcinogenesis. This promising model could facilitate the study of host-pathogen interactions and the investigation of the process involved in Cryptosporidium-induced cell transformation.

[Comparison of immunochromatographic tests Actim(®) Prom and Amnioquick(®) Duo for the rapid detection of premature rupture of membranes]. / Comparaison des tests immunochromatographiques Actim(®) Prom et Amnioquick(®) Duo pour le dépistage rapide de la rupture prématurée des membranes.

Premature rupture of membranes (PRM) affects 5 to 15% of pregnancies, leading to prematurity and neonatal infection. PRM can be identified by through various amniotic fluid proteins in vaginal secretions. The aim of this study is to compare two immunochromatographic tests based on the detection of insulin-like growth factor binding protein (IGFBP-1) and alpha-foeto protein (AFP) for one of the two tests in cervico-vaginal secretions. Two tests, Actim(®) Prom and Amnioquick(®) Duo were performed on 80 pregnant women with suspected PRM. Amnioquick(®) Duo allows the simultaneous detection of IGFBP-1 and AFP with an automated incubation and reading. The number of positive results is similar (Khi-deux = 0.173, p = 0.6773) for IGFBP-1 between the two tests and there is a good agreement (K = 0.621), with a proportion of negative results of 86%. The number of positive results for AFP is more important in comparison to IGFBP-1. Results positive/positive (Actim(®) Prom/Amnioquick(®)) for IGFBP-1 seems to be related to the time when tests have been performed, that is to say in the last weeks of pregnancy. In conclusion, both tests have similar performance, but there is a risk of false positive results with AFP, this can be explained by the presence of non-visible blood in samples. An automated incubation and reading allows a good reproducibility. Moreover, the computer data storage improve the post-analytical quality.

Correlation between the anti-virus-induced cytopathic effect activity of interferon-alpha subtypes and induction of MxA protein in vitro.

There are several interferon-alpha (IFN-alpha) subtypes. Mechanism of disparity in biological effects among members of IFN-alpha subtypes remains unexplained. Biological activity of IFN-alpha is mediated in part by induction of intracellular antiviral proteins. We studied whether differences in biologic effects of IFN-alpha subtypes may rely on their antiviral protein inducing effect. Intracellular induction of MxA protein and anti-virus-induced cytopathic effect (CPE) activity of 11 IFN-alpha subtypes in human amnion WISH cells have been studied. MxA protein quantitation in cell lysates was performed by immunochemiluminescence assay and anti-virus-induced CPE activity was assessed by protection against vesicular stomatitis virus (VSV)-induced CPE. Range of MxA values was high when cells were treated with 10 and 100 IU/ml of each IFN-alpha subtype. Levels of MxA correlated with anti-VSV-induced CPE obtained with 10 IU/ml IFN-alpha subtype. Together our data show a disparity in MxA-inducing activity of IFN-alpha subtypes and suggest that differences in anti-VSV-induced CPE of IFN-alpha subtypes in WISH cells can be related to their different ability to induce MxA.

Study of serum factors potentially involved in the pathogenesis of heterotopic bone formation after severe brain injury.

OBJECTIVE: To look for serum factors detectable early after head injury and predictive of heterotopic bone formation. PATIENTS AND METHODS: In this prospective study of a homogeneous population of 31 men with severe brain injury, blood samples were obtained 3 months after the accident, and levels of serum factors influenced by bone metabolism were compared between patients with and without heterotopic bone formation. As extensive fractures can influence serum factors, the patients without heterotopic bone formation were divided into two groups based on whether they had major fractures. Radionuclide bone scanning was used to validate patient classification. RESULTS: The group with heterotopic bone formation had significantly higher serum alkaline phosphatase levels (P < 0.01) and significantly lower serum leptin levels (P < 0.01), as compared to the other two groups. Body mass index and serum creatinine were comparable in the three groups. CONCLUSION: Leptin may be associated with the development of heterotopic bone formation. The antiosteogenic effect of leptin mediated by hypothalamic neurons may be impaired by hypothalamic damage related to severe brain injury.