Abstract: Staining of spermatozoa with the fluorescein-Val-Ala-Asp-fluoromethylketone has already been performed on ejaculated sperm samples, using fluorescence microscopy (FM) or flow cytometry (FCM) in order to score activated caspases. This assay may help in assessing apoptosis and its role in male fertility. The present study compares the above two techniques in order to adopt the most accurate method for detection in human frozen-thawed testicular, epididymal and ejaculated spermatozoa. The analyses were carried out on frozen/thawed testicular (n = 14), epididymal (n = 8) and ejaculated (n = 10) sperm samples. Activated caspases were detected in living spermatozoa using fluorescein-labelled inhibitors of poly-caspases (FLICA). For the measurements by FM, the same-observer and different-observer reliability were assessed in testicular and epididymal sperm samples. The inter-method (FM and FCM) reliability was assessed both in epididymal and ejaculated sperm samples. The reliability was evaluated by intraclass correlation coefficient (ICC) and the differences between paired measurements from the same sample were tested by Wilcoxon test for matched pairs. For the same-observer and the different-observer data, the ICC were 0.980 and 0.986. In testicular suspensions, the presence of different types of germinal and somatic cells hampers the differentiation of stained spermatozoa by FCM. For the inter-method reliability, the ICC was 0.903. A lower proportion of the viable spermatozoa stained with FLICA was observed by using FM (-6.60 +/- 7.38 %, mean +/- SD; p = 0.003) compared with FCM. To measure the proportion of spermatozoa with activated caspases by this test, FM is a highly accurate and reliable method whatever the sperm origin (ejaculate, epididymis, and testis). FCM cannot be used for testicular samples but seems to be more appropriate for analysis of epididymal and ejaculated sperm samples. The systematic lower proportion by FM in measuring the proportion of stained viable spermatozoa with FLICA involves that only the data measured by the same method (FM or FCM) may be compared.

Abstract: Due to its extensive production and application, the toxicity of chloracetanilide herbicide alachlor[2-chloro-2',6'-diethyl-N-(methoxymethyl)-acetanilide] should be evaluated to establish minimum effects. In this study, we have examined the in vitro effects of alachlor on human sperm motion using a computer-assisted sperm analyser (CASA). An evaluation of both reactive oxygen species (ROS) and markers of apoptosis was also performed to investigate the mechanism by which alachlor modifies the sperm movement. After exposure up to 2 h to alachlor (0, 0.18, 0.37, 0.90 and 1.85 mM), the percentage of viable, motile spermatozoa and sperm velocities were concentration and/or time dependently decreased. The most sensitive parameters were progressive motility, mean average path velocity and mean straight velocity. Alachlor (1.85 mM) induced an increase in ROS production. A decrease of mitochondrial membrane potential (DeltaPsi(m)), an increase of both phosphatidylserine (PS) externalization and DNA fragmentation, which were concentration and/or time dependent, were also observed. It is possible that toxic effects of alachlor result in an oxidative stress which could act as a mediator of apoptosis. Alachlor could also contribute to some hypofertility cases.

Abstract: OBJECTIVE: To assess the role of age in the results of bilateral deep brain stimulation in the subthalamic nucleus (DBS-STN), we carried out a study of two groups of patients regarding age at time of surgery. METHODS: We compared, up to 2 years after surgery, the clinical effects, safety, and quality of life in parkinsonian patients younger than 65 years old (young patients) vs parkinsonian patients 65 years old or older (old patients). RESULTS: The mean age was 57.4 +/- 4.9 years (n = 53) in young patients and 68.8 +/- 2.8 years (n = 34) in old patients. A dramatic improvement in motor complications was equally observed in both groups of patients. There was no significant difference between the groups regarding acute effects of DBS-STN on the motor score of the Unified Parkinson's Disease Rating Scale part III (UPDRS III). Time course evolution of UPDRS the motor score (p < 0.0001) and axial score (p = 0.0001) assessed postoperatively in "on" medication and "on" stimulation conditions appeared worse in old patients as compared to young patients. Improvement in the Schwab and England Scale score was better in young patients in "on" (p < 0.0003) and "off" state (p < 0.001). Quality of life assessed with the 39-item Parkinson's Disease Questionnaire showed an improvement in subscales evaluating mobility (p < 0.0001), activities of daily life (p < 0.0001), emotion and stigma (p = 0.0004), cognition (p < 0.0074), and communication (p = 0.0029) in young patients as compared to old patients. Side effects were similar in the two groups. CONCLUSIONS: Although deep brain stimulation in the subthalamic nucleus reduces motor complications equally in both groups of patients, postoperative quality of life improved only in young patients.

Abstract: The mechanism of action of acetaminophen is currently widely discussed. Direct inhibition of cyclooxygenase isoforms remains the commonly advanced hypothesis. We combined behavioral studies with molecular techniques to investigate the mechanism of action of acetaminophen in a model of tonic pain in rats. We show that acetaminophen indirectly stimulates spinal 5-hydroxytryptamine (5-HT)1A receptors in the formalin test, thereby increasing transcript and protein levels of low-affinity neurotrophin receptor, insulin-like growth factor-1 (IGF-1) receptor alpha subunit, and growth hormone receptor and reducing the amount of somatostatin 3 receptor (sst3R) mRNA. Those cellular events seem to be important for the antinociceptive activity of acetaminophen. Indeed, down-regulation of sst3R mRNA depends on acetaminophen-elicited, 5-HT1A receptor-dependent increase in neuronal extracellular signal-regulated kinase 1/2 (ERK1/2) activities that mediate antinociception. In addition, spinal growth hormone (GH) and IGF-1 receptors would also be involved in the antinociceptive activity of the analgesic at different degrees. Our results show the involvement of specific 5-HT1A receptor-dependent cellular events in acetaminophen-produced antinociception and consequently indicate that inhibition of cyclooxygenase activities is not the exclusive mechanism involved. Furthermore, we propose that the mechanisms of 5-HT1A receptor-elicited antinociception and the role of the spinal ERK1/2 pathway in nociception are more intricate than suspected so far and that the GH/IGF-1 axis is an interesting new player in the regulation of spinal nociception.

Abstract: OBJECTIVES: NOD2/CARD15 is a susceptibility gene for Crohn's disease (CD). It is also involved, via different mutations, in the Blau syndrome. The syndrome of aseptic abscesses (AA) is characterized by visceral sterile collections of mature neutrophils that do not respond to antibiotics but regress quickly with corticosteroids. It is associated in two cases out of three with inflammatory bowel disease (IBD), and in particular with CD. We wanted to assess if changes on gene NOD2/CARD15 could contribute to the development of AA in patients with and without IBD. METHODS: Seventeen unrelated patients with AA from the French national register were genotyped for c.802C>T (p.Pro268Ser) and the three main CD associated variants, c.2104C>T (p.Arg702Trp), c.2722G>C (p.Gly908Arg) and c.3019_3020insC (p.Leu1007fsX1008), and 16 were screened for the 11 coding exons of NOD2/CARD15. RESULTS: The main variants associated with CD were found at a similar frequency in patients free of IBD and in those with CD. There was no significant difference in the main variants between patients with CD and those without IBD in our study and patients with CD and controls, respectively, from a large study of an ethnically similar population. No rare variant was found. A significant association between carriers of the silent variant c.1377 C>T and markers of severity of AA was observed. CONCLUSIONS: These results suggest that the emergence of AA is not closely related to gene NOD2/CARD15. NOD2/CARD15 and other susceptibility genes might enhance the expression of AA as the result of a combination of polymorphisms.

Abstract: In this article, we briefly review the concept of brain mapping in stereotactic surgery taking into account recent advances in stereotactic imaging. The gold standard continues to rely on probabilistic and indirect targeting, relative to a stereotactic reference, i.e., mostly the anterior (AC) and the posterior (PC) commissures. The theoretical position of a target defined on an atlas is transposed into the stereotactic space of a patient's brain; final positioning depends on electrophysiological analysis. The method is also used to analyze final electrode or lesion position for a patient or group of patients, by projection on an atlas. Limitations are precision of definition of the AC-PC line, probabilistic location and reliability of the electrophysiological guidance. Advances in MR imaging, as from 1.5-T machines, make stereotactic references no longer mandatory and allow an anatomic mapping based on an individual patient's brain. Direct targeting is enabled by high-quality images, an advanced anatomic knowledge and dedicated surgical software. Labeling associated with manual segmentation can help for the position analysis along non-conventional, interpolated planes. Analysis of final electrode or lesion position, for a patient or group of patients, could benefit from the concept of membership, the attribution of a weighted membership degree to a contact or a structure according to its level of involvement. In the future, more powerful MRI machines, diffusion tensor imaging, tractography and computational modeling will further the understanding of anatomy and deep brain stimulation effects.

Abstract: This prospective, single centre, randomised, cross-over study compares patient tolerance of the facemask and mouthpiece for delivery of non-invasive ventilation in an intensive care unit. Twenty-seven patients with acute respiratory failure were scheduled for two 45-min sessions of non-invasive ventilation with facemask and mouthpiece. The order of the sessions was chosen at random. Nurses and patients assessed the tolerance of both techniques using a visual analogue scale. The time spent by nurses and the changes in respiratory parameters were recorded. The facemask was better tolerated than the mouthpiece; all the cases of non-invasive ventilation withdrawal (n = 5) occurred with mouthpieces (p = 0.026). Less nursing time was required using the facemask for the 22 patients who underwent both procedures (p = 0.01). However, the difference in tolerance scores was not significant. Non-invasive ventilation with both facemask and mouthpiece improved the P(a)o(2)/F(i)o(2) ratio, increased the pH and decreased the P(a)co(2). Only non-invasive ventilation with the facemask lowered the respiratory rate. The facemask appears to be a better initial choice for non-invasive ventilation when compared to mouthpiece, but both can be effective.

Abstract: INTRODUCTION: The SPOT epidemiologic survey assessed the results of long-term cholesterol-lowering therapy in France and the reasons why patients do or do not meet their therapeutic objectives. OBJECTIVE: To assess the guidelines on which doctors base their practices for cholesterol-lowering therapy and make suggestions to improve the match between recommendations and practices. METHODS: LDL cholesterol was measured in 2479 French outpatients in 2003 and classified according to whether it met therapeutic objectives of 3 different scales: the official French (AFSSAPS) recommendations issued in 2000, the physician's opinion of the ideal level for the patient, and the physician's goal level for the patient. Agreement between these 3 criteria was analyzed. RESULTS: French doctors used guidelines stricter than the official recommendations to assess the achievement of lipid-lowering objectives. DISCUSSION: Physicians rely more on American than French guidelines. They also considered the patient's history and clinical condition in setting individual goals.

Abstract: Symptoms of Parkinson's disease can be relieved through deep brain stimulation. This neurosurgical technique relies on high precision positioning of electrodes in specific areas of the basal ganglia and the thalamus. In order to identify these anatomical targets, which are located deep within the brain, we developed a semi-automated method of image analysis, based on data fusion. Information provided by both anatomical magnetic resonance images and expert knowledge is managed in a common possibilistic frame, using a fuzzy logic approach. More specifically, a graph-based virtual atlas modeling theoretical anatomical knowledge is matched to the image data from each patient, through a research algorithm (or strategy) which simultaneously computes an estimation of the location of every structures, thus assisting the neurosurgeon in defining the optimal target. The method was tested on 10 images, with promising results. Location and segmentation results were statistically assessed, opening perspectives for enhancements.

Abstract: The impact of motor complications of Parkinson's disease (PD), especially levodopa-induced dyskinesias, on quality of life (QL) was studied in 143 patients with PD. All were evaluated on the Hoehn and Yahr (H&Y) scale, and the Motor part of the Unified Parkinson's Disease Rating Scale (UPDRS). Motor complications were analyzed using the UPDRS Parts IV(A) and IV(B) and the Abnormal Involuntary Movement Scale. A specific Parkinson's disease quality of life questionnaire (39-item version, PDQ-39) was used. Motor complications significantly worsened the PDQ-39 Summary Index (PDQ-SI) of patients with PD. The dimensions of Mobility, Activities of Daily Living, Stigma, and Communication were the most strongly affected. "Peak dose" dyskinesia decreased Mobility, Emotional Well-Being, and Cognition, whereas biphasic dyskinesia affected Mobility, Stigma, Communication, and Activities of Daily Living. Morning akinesia, end-of-dose fluctuations, and "unpredictable offs" decreased QL on the dimensions of Mobility, Activities of Daily Living, Stigma, and Communication. Nocturnal akinesia led to a deterioration of all dimensions of the PDQ-39. Thus, motor complications and especially nocturnal akinesia and biphasic dyskinesias worsened the QL of PD patients.

Abstract: Airway inflammation is the principal abnormality in asthma and many other respiratory diseases. Eosinophils are the cells primarily involved in this process. The aim of this study was to analyze sequential changes in urinary eosinophil protein X (EPX) a biological marker of eosinophil activation in asthmatic children and chronic coughers, and to confirm the importance of such changes in evaluating the inflammatory process once regular treatment was initiated. Eighty-eight asthmatic children (AC), 33 children with chronic cough (CC), and 34 control children were included in the study. All those with respiratory disease underwent allergy tests (serum total IgE, serum-specific IgE for common allergens, peripheral blood eosinophil (PBE), and skin prick tests) and a pulmonary function test (PFT), and had chest X-ray and serum eosinophil cationic protein (s-ECP) and urinary EPX assays. All subjects attended the outpatient clinic every 3 months, irrespective of the treatment prescribed following inclusion in this investigation. At baseline, urinary EPX concentrations were higher in children with asthma and those with chronic cough than in controls (mean 171.1 and 131.3, respectively, vs. 60.2 microg/mmol creatinine, P < 0.001). CC children had lower eosinophil counts (0.25 vs. 0.39 x 10(9)/L, P < 0.02) than those with asthma. There was no significant difference between the AC and CC groups in urinary EPX and s-ECP levels. s-ECP concentrations were significantly higher (P < 0.01) in atopic vs. nonatopic patients (44 vs. 29.9 ng/mL), but no significant difference was observed for urinary EPX. Concentrations of urinary EPX were significantly correlated with s-ECP levels (r = 0.24, P < 0.025) and with PBE (r = 0.38, P < 0.01). No correlation was found between urinary EPX values and PFT results. In AC receiving inhaled steroids after the start of the study, there was a significant reduction after 3 months in urinary EPX (-54, P < 0.02). In contrast, there was no significant change in PBE levels. Urinary EPX concentrations are sensitive, noninvasive technique that could be useful to the clinician in the evaluation of manifestations of airway inflammation.