Cytomorphometric Evaluation of Oral Mucosa of Patients with Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a rare genetic disorder in which patients progressive muscle deterioration and low life expectancy. Since DMD has no cure, treatment slows the deterioration process using corticosteroids and other medications, including oral bisphosphonates (BP). These drugs can lead to toxicities in both soft and hard tissues. The objective of this study was to present cytological changes in DMD patients, comparing to young, healthy individuals. Cytological smears were obtained from buccal mucosa from nine DMD patients and five healthy patients. The Papanicolaou technique was used to stain the slides. Images were captured using a photomicroscope, and 50 clearly defined cells were selected. The size and ratio between the nucleus and cytoplasmic areas (NA/CA) diameter were measured using a computer program. All the DMD patients presented a statistical difference (p < 0.0001) in the size of the cytoplasm (0.60 ± 0.33 µm) and the NA/CA ratio (0.17 ± 0.07 µm) was seen when compared to the control group (cytoplasm 0.60 ± 0.15 µm and nucleus 0.14 ± 0.05 µm). Furthermore, in one DMD in use of BP who presented Medication-Related Osteonecrosis of the Jaw (MRONJ), presented higher differences in the cytoplasmic (1.20 ± 0.65 µm) and nuclear (0.20 ± 0.12 µm) sizes (p < 0.0001) compared with all others DMD patients. These results suggest that the drugs used or the systemic condition of individuals with DMD may contribute to these changes.

Mandatory First Impressions: Happy Expressions Increase Trustworthiness Ratings of Subsequent Neutral Images.

First impressions of traits are formed rapidly and nonconsciously, suggesting an automatic process. We examined whether first impressions of trustworthiness are mandatory, another component of automaticity in face processing. In Experiment 1a, participants rated faces displaying subtle happy, subtle angry, and neutral expressions on trustworthiness. Happy faces were rated as more trustworthy than neutral faces; angry faces were rated as less trustworthy. In Experiment 1b, participants learned eight identities, half showing subtle happy and half showing subtle angry expressions. They then rated neutral images of these same identities (plus four novel neutral faces) on trustworthiness. Multilevel modeling analyses showed that identities previously shown with subtle expressions of happiness were rated as more trustworthy than novel identities. There was no effect of previously seen subtle angry expressions on ratings of trustworthiness. Mandatory first impressions based on subtle facial expressions were also reflected in two ratings designed to assess real-world outcomes. Participants indicated that they were more likely to vote for identities that had posed happy expressions and more likely to loan them money. These findings demonstrate that first impressions of trustworthiness based on previously seen subtle happy, but not angry, expressions are mandatory and are likely to have behavioral consequences.

Shiga-toxigenic Escherichia coli from animal food sources in Mauritius: Prevalence, serogroup diversity and virulence profiles.

Shiga-toxigenic Escherichia coli (STEC) are important human pathogens associated with diarrhea and in some cases haemorrhagic colitis. Contaminated food derived from cattle and wildlife species are often associated with disease outbreaks. In this study, we report the prevalence, serogroup diversity and virulence profiles of STEC strains derived from cattle, rusa deer and pig. Of the 422 samples analyzed, STEC were detected in 40% (80/200) of cattle, 27.0% (33/122) of deer and 13.0% (13/100) of pigs. STEC isolates belonged to 38 O-serogroups whereby 5.2% (24/462) of the isolates belonged to clinically important EHEC-7 serogroups: O26 (n = 2), O103 (n = 1), O145 (n = 3) and O157 (n = 18). Fourteen serogroups (O26, O51, O84, O91, O100, O104, O110, O117, O145, O146, O156, O157, O177 and ONT) displayed multiple virulence profiles. We also identified two serovars (O117 and O119) in deer which are not well-documented in epidemiological surveys. 73.7% (28/38) of recovered O-serogroups are known to be associated with serious human illnesses including haemolytic uremic syndrome (HUS) and bloody diarrhea. STEC isolates harboring single genotypes stx1, stx2, eae and hlyA accounted for 3.0% (14/462), 9.1% (42/462), 47.6% (220/462) and 1.7% (8/462) of all STEC isolates screened, respectively. Virulence combinations stx1 and stx2 were harboured by 1.3% of isolates while strains with genetic profiles eae/hlyA were the second most prevalent amongst STEC isolates. The full known virulent genotypes (stx2/eae, stx1/stx2/eae, stx1/stx2/hlyA and stx2/eae/hlyA) were present in 22 of the 462 STEC strains. A total of 10 different virulence patterns were recovered amongst animal species. Phylogeny of the gnd gene showed that amongst STEC strains, serovar O100 outlined the main cluster. Fourteen (n = 14) different sequence types (STs) were identified from a panel of twenty (n = 20) STEC isolates. One of the isolate (PG007B) possessed a unique ST (adk 10, fumC 693, gyrB 4, icd 1, mdh 8, purA 8, recA 2) that could not be assigned using MLST databases. None of the ST's recovered in deer were observed in domestic species. Our findings shows that food associated animals found on the tropical island of Mauritius carry a diversity of STEC strains with many serovars known to be associated with human disease. This report indicates that increased awareness, surveillance and hygienic attention at critical stages of the human food chain are warranted.

A cluster of Chlamydia serpentis cases in captive snakes.

Following the occurrence of sudden death cases in a zoo reptile collection, histological analyses conducted on tissues from two common adders suggested an infection due to Chlamydia. The survey was extended to 22 individual snakes from the same collection and a PCR analysis targeting a conserved gene in Chlamydiaceae revealed bacterial shedding in six of them. The infection resolved spontaneously in one snake whereas another one succumbed one month later. The antibiotic treatment administered (marbofloxacin) to the remaining four PCR positive animals stopped the mortalities and the shedding. Analysis of the 16S and 23S ribosomal gene sequences identified C. serpentis, a recently described novel chlamydial species in snakes. A PCR tool for a quick and specific identification of this new chlamydial species was developed in this study.

[Development and first results of fast and cost-effective examination methods for an ophthalmological screening within the National Cohort]. / Entwicklung und erste Anwendungserfahrungen einer schnellen und kostengünstigen ophthalmologischen Untersuchung für die Nationale Kohorte.

BACKGROUND: For an implemented ophthalmological screening within a German long-term cohort study (National Cohort) simple and effective methods for an examination of visual acuity and for non-mydriatic retina photografies should be evaluated. Furthermore standard operating-procedures (SOP) should be developed. METHODS: In the years 2011 and 2012 pinhole visual acuity measurements and automated retina photographies (DRS, CenterVue S. p. a., Padua, Italy) were made at three different epidemiological study centers within Germany. Furthermore, anterior segment images were taken by the camera. Standard operating procedures (SOP) regarding the ophthalmological screening were developed and evaluated within the study. The main question was whether it is possible to implement the screening methods within the National Cohort. Further main outcomes were quality and interpretability of the taken images. RESULTS: 457 subjects (914 eyes) were examined within the investigation. Median VA was 0.8 for right and left eyes (p > 0.42). Image quality of the photographies was good in 491 cases (54 %), fair in 239 cases (26 %) and bad in 179 cases (20 %). The usability of the images was without limitations in 686 cases (75 %), limited in 152 cases (17 %) and not given in 71 cases (8 %). Increasing age of the subjects was slightly correlated with decreasing image quality (r = 0.26) and decreasing image usability (r = 0.2). Anterior segment photographies were usable in 176 eyes (56 %). CONCLUSION: The developed screening method fulfilled the specifications of the National Cohort. The used pinhole visual acuity examination was fast and cheap. Image quality and usability of the retina photographies could be improved with prolonged pupil recovery times. The quality of the anterior segment images could not fulfill the expectations and were taken out of the further examinations of the ophthalmological screening. The written SOP showed good acceptance within the investigators' daily routine. The ophthalmological screening within the National Cohort generates information (e. g., pathologies of the vessels or of the retina) which are useful not only from an ophthalmological point of view.

Stimulation of the human RAD51 nucleofilament restricts HIV-1 integration in vitro and in infected cells.

Stable HIV-1 replication requires the DNA repair of the integration locus catalyzed by cellular factors. The human RAD51 (hRAD51) protein plays a major role in homologous recombination (HR) DNA repair and was previously shown to interact with HIV-1 integrase (IN) and inhibit its activity. Here we determined the molecular mechanism of inhibition of IN. Our standard in vitro integration assays performed under various conditions promoting or inhibiting hRAD51 activity demonstrated that the formation of an active hRAD51 nucleofilament is required for optimal inhibition involving an IN-DNA complex dissociation mechanism. Furthermore we show that this inhibition mechanism can be promoted in HIV-1-infected cells by chemical stimulation of the endogenous hRAD51 protein. This hRAD51 stimulation induced both an enhancement of the endogenous DNA repair process and the inhibition of the integration step. Elucidation of this molecular mechanism leading to the restriction of viral proliferation paves the way to a new concept of antiretroviral therapy based on the enhancement of endogenous hRAD51 recombination activity and highlights the functional interaction between HIV-1 IN and hRAD51.

Characterization of genetic diversity of Bacillus anthracis in France by using high-resolution melting assays and multilocus variable-number tandem-repeat analysis.

Using high-resolution melting (HRM) analysis, we developed a cost-effective method to genotype a set of 13 phylogenetically informative single-nucleotide polymorphisms (SNPs) within the genome of Bacillus anthracis. SNP discrimination assays were performed in monoplex or duplex and applied to 100 B. anthracis isolates collected in France from 1953 to 2009 and a few reference strains. HRM provided a reliable and cheap alternative to subtype B. anthracis into one of the 12 major sublineages or subgroups. All strains could be correctly positioned on the canonical SNP (canSNP) phylogenetic tree, except the divergent Pasteur vaccine strain ATCC 4229. We detected the cooccurrence of three canSNP subgroups in France. The dominant B.Br.CNEVA sublineage was found to be prevalent in the Alps, the Pyrenees, the Auvergne region, and the Saône-et-Loire department. Strains affiliated with the A.Br.008/009 subgroup were observed throughout most of the country. The minor A.Br.001/002 subgroup was restricted to northeastern France. Multiple-locus variable-number tandem-repeat analysis using 24 markers further resolved French strains into 60 unique profiles and identified some regional patterns. Diversity found within the A.Br.008/009 and B.Br.CNEVA subgroups suggests that these represent old, ecologically established clades in France. Phylogenetic relationships with strains from other parts of the world are discussed.

CSF neurofilament protein analysis in the differential diagnosis of ALS.

BACKGROUND: Cerebrospinal fluid (CSF) biomarkers have been studied to differentiate between patients with ALS and neurological controls, but not in comparison to clinically more relevant disorders mimicking ALS. METHODS: In this retrospective study, CSF concentrations of various brain-specific proteins were analyzed in patients with ALS (n = 32) and ALS-mimic disorders (n = 26). RESULTS: CSF concentrations of neurofilament light (NFL) and heavy chain (NFHp35), but not other brain-specific proteins, were significantly higher in patients with ALS than in patients with an ALS-mimic disorder, however with maximum sensitivity or specificity of 80%. The mean CSF level of NFHp35 was 781 ng/L in the ALS group vs. 338 ng/L in the ALS-mimic disorders group and for NFL the mean CSF levels were 62 ng/L vs. 24 ng/L. CONCLUSION: Although CSF concentrations of NFL and NFHp35 are higher in patients with ALS, the diagnostic accuracy for differentiating ALS from ALS-mimic disorders seems insufficient. Our results suggest that, in the clinical work-up of patients suspected of ALS, application of CSF analysis alone is limited but may have potential in combination with other clinical and electrophysiological markers.

Pleural plaques and asbestosis: dose- and time-response relationships based on HRCT data.

Most studies on asbestos-related diseases are based on chest radiographs, and dose-response relationships are still controversial. The aim of this study was to describe the most relevant parameters of asbestos exposure linked to pleural plaques and asbestosis diagnosed by high-resolution computed tomography (HRCT). A large screening programme including systematic HRCT examinations was organised from 2003 to 2005 in France for formerly asbestos-exposed workers. The time since first exposure (TSFE), level, duration and cumulative exposure to asbestos were used in adjusted unconditional logistic regression to model the relationships of the two diseases. Analysis of a population of 5,545 subjects demonstrated that TSFE (p<0.0001) and cumulative exposure (p = 0.02) (or level, depending on the models used), were independently associated with the frequency of pleural plaques. Only cumulative exposure (p<0.0001) or level of exposure (p = 0.02) were significantly associated with asbestosis. All trend tests were significant for these parameters. To date and to our knowledge, this study is the largest programme based on HRCT for the screening of asbestos-exposed subjects. Both time-response and dose-response relationships were demonstrated for pleural plaques, while only dose-response relationships were demonstrated for asbestosis. These parameters must be included in the definition of high-risk populations for HRCT-based screening programmes.

Echocardiographic features, mortality, and adrenal function in patients with cirrhosis and septic shock.

OBJECTIVES: Cirrhosis of the liver is associated with an increased susceptibility to bacterial infections capable of causing septic shock and with a basal hyperdynamic circulatory state. The primary objective of this study was to delineate the echocardiographic characteristics and outcomes of septic shock in patients with liver cirrhosis. The secondary objective was to determine whether adrenal insufficiency, which may contribute to hyperdynamic syndrome, was more marked in patients with cirrhosis than in other patients with septic shock. DESIGN: Prospective single-center cohort study. PATIENTS AND METHODS: Thirty-four patients admitted to the intensive care unit (ICU) for septic shocks were included, 14 with and 20 without liver cirrhosis. Echocardiography was performed within the first 24 h to measure the cardiac index (CI), systolic index (SI), and left ventricular ejection fraction (LVEF). A Synacthen test was performed. RESULTS: Patients with cirrhosis had higher values for the CI (3.69+/-1.0 vs. 2.86+/-0.8 l/min/m(2); P=0.02), SI (37.5+/-8 vs. 32.4+/-7 ml/m(2); P=0.04), and LVEF (67+/-7 vs. 55.9+/-12%; P=0.005). ICU mortality was 53% overall, 64% in patients with cirrhosis, and 45% in patients without cirrhosis (P=0.27). Serum cortisol levels under basal conditions (H0) and after stimulation (H1) showed no significant differences between patients with and without cirrhosis. The proportion of patients with no response to Synacthen was 77% among patients with cirrhosis and 50% among patients without cirrhosis (P=0.18). CONCLUSION: In a population with septic shock, left ventricular function was more hyperdynamic in the subset with cirrhosis. Relative adrenal insufficiency occurred in similar proportions of patients with and without cirrhosis.

Diagnostic accuracy of ELISA and xMAP technology for analysis of amyloid beta(42) and tau proteins.

BACKGROUND: Cerebrospinal fluid (CSF) concentrations of amyloid beta(42) (Abeta(42)) peptides and tau proteins may serve as biomarkers for Alzheimer disease (AD). Recently, the xMAP technology has been introduced as an alternative to ELISA for measurement of these markers. METHODS: We used xMAP assays and ELISA to analyze CSF concentrations of Abeta(42), total tau (t-tau), and tau phosphorylated at threonine 181 (p-tau(181)) in samples from 69 patients with Alzheimer disease, 26 patients with vascular dementia, and 55 controls without neurological disorders. RESULTS: High CV values (>28%) for the ratio of xMAP:ELISA were observed for each biomarker, indicating that a constant correction factor cannot be applied to recalculate xMAP results into ELISA results. When a combination of CSF markers was used, the sensitivity, specificity, and area under the ROC curves for xMAP assays and ELISAs were not significantly different in differentiating AD patients from vascular dementia patients and controls. CONCLUSIONS: A constant conversion factor cannot be used successfully to recalculate results obtained with xMAP assays to those from the ELISAs. With the use of analysis of a combination of Abeta(42), t-tau, and p-tau in CSF, however, differentiation of clinical groups is equivalent when either xMAP technology or conventional ELISA is used.

[Interest of the brain natriuretic peptide as a marker of acute cor pulmonale in acute respiratory distress syndrome]. / Analyse du peptide natriurétique de type B comme marqueur du coeur pulmonaire aigu dans le syndrome de détresse respiratoire aiguë.

OBJECTIVE: The aim of this study was to evaluate the accuracy of the BNP as a marker of acute cor pulmonale in patients with ARDS. STUDY DESIGN: Prospective clinical trial. PATIENTS AND METHODS: At day 2 or 3 after the onset of the ARDS, an echocardiography was performed. Patients with left ventricular dysfunction were excluded. Right ventricular area (RVA) and RVA/LVA ratio were measured. ACP was defined as RVA/LVA > 0.6 associated with septal dyskinesia. Simultaneously, 5 ml of blood was collected for BNP measurement. RESULTS: 26 patients were studied. BNP levels were higher in 10 patients with ACP: 585.5 [189-4830] vs 145.5 [36.5-346] pg/ml (P=0.01) but in those with creatinine clearance < 90 ml/min: 602 [331-3530] vs 125 [39-189] pg/ml (P=0.007). BNP was correlated with RVA (r=0.5; p=0.01), RVA/LVA ratio (r=0.61; p=0.001), sPAP (r=0.58; p=0.002) and with age, cardiac index and creatinine clearance (r=0.61; p=0.001). In multivariate analysis, BNP was only correlated with creatinine clearance (p=0.03), and RVA (p=0.06). CONCLUSION: In ARDS patients without left ventricular dysfunction, BNP level is more elevated in patients with acute cor pulmonale than patients without cor pulmonale.

B-type natriuretic peptide (BNP) and N-terminal-proBNP for heart failure diagnosis in shock or acute respiratory distress.

BACKGROUND: Plasma B-type natriuretic peptide (BNP) assay is recommended as a diagnostic tool in emergency-room patients with acute dyspnea. In the intensive care unit (ICU), the utility of this peptide remains a matter of debate. The objectives of this study were to determine whether cut-off values for BNP and N-terminal-proBNP (NT-proBNP) reliably diagnosed right and/or left ventricular failure in patients with shock or acute respiratory distress, and whether non-cardiac factors led to an increase in these markers. METHODS: Plasma BNP and NT-proBNP levels and echocardiographic parameters of cardiac dysfunction were determined in 41 patients within 24 h of the onset of shock or acute respiratory distress. RESULTS: BNP and NT-proBNP levels were higher in the 25 patients with heart failure than in the other 16 patients: 491.7 +/- 418 pg/ml vs. 144.3 +/- 128 pg/ml and 2874.4 +/- 2929 pg/ml vs. 762.7 +/- 1128 pg/ml, respectively (P < 0.05). In the diagnosis of cardiac dysfunction, BNP > 221 pg/ml and NT-proBNP > 443 pg/ml had 68% and 84% sensitivity, respectively, and 88% and 75% specificity, respectively, but there was a substantial overlap of BNP and NT-proBNP values between patients with and without heart failure. BNP and NT-proBNP were elevated, but not significantly, in patients with isolated right ventricular dysfunction. Patients with renal dysfunction and normal heart function had significantly higher levels of BNP (258.6 +/- 144 pg/ml vs. 92.4 +/- 84 pg/ml) and NT-proBNP (2049 +/- 1320 pg/ml vs. 118 +/- 104 pg/ml) than patients without renal dysfunction. CONCLUSION: Both BNP and NT-proBNP can help in the diagnosis of cardiac dysfunction in ICU patients, but cannot replace echocardiography. An elevated BNP or NT-proBNP level merely indicates the presence of a 'cardiorenal distress' and should prompt further investigation.

[Clinical practices of analgesia for invasive procedures in critically ill sedated patients in Ile-de-France: a phone survey]. / Enquête téléphonique sur les pratiques cliniques d'analgésie pour les actes invasifs chez les patients sédatés en réanimation en Ile-de-France.

To assess the practice of analgesia for invasive procedures in critically ill sedated patient in Ile-de-France (French area including Paris). Observational study: phone survey using a standard questionnaire. Only one senior physician in each of 30 intensive care unit (ICU) was questioned. Baseline sedation included systematic analgesia with narcotics in all ICUs. Only 4 physicians declared using a specific pain scale for sedated patients. Only 3 ICUs used written protocols. Procedures, which were thought to be most invasive (catheterization, pleural drainage, fibroscopy) were in most cases preceded by analgesia, but this was seldom the case for less painful events (venous or arterial puncture, tracheal suctioning). Specific pain scales are still underused. In contrast with current guidelines, analgesia for invasive procedures is not systematic but depends on subjective opinions.

[Color M-mode Doppler for postoperative estimation of pulmonary capillary wedge pressure]. / Estimation postopératoire de la pression artérielle pulmonaire d'occlusion par étude Doppler, en mode M couleur, du flux transmitral.

OBJECTIVE: The aim of this study was to evaluate the accuracy of the combined index E/Vp (peak E velocity combined with color M-mode color Doppler flow propagation) for estimating pulmonary capillary wedge pressure, in post cardiac surgery patients. STUDY DESIGN: Prospective clinical trial. PATIENTS AND METHODS: In post cardiac surgery patients (D1), we have measured with transthoracic echocardiography peak early E transmitral pulsed Doppler velocities and color M-mode Doppler flow propagation velocity (Vp). The E/Vp ratio was compared with pulmonary capillary wedge pressure (PAPO) obtained simultaneously. RESULTS: Thirty eight patients were studied. The coefficient of correlation between PAPO and E/Vp was r = 0.71 (p < 0.0001). The sensitivity and the specificity of E/Vp > 1.5 for prediction of PAPO > 15 mmHg were 79% and 79% respectively. CONCLUSION: In post cardiac surgery patients, PAPO can be reasonably estimated by measuring the ratio E/Vp obtained with Doppler echocardiography.

[Value of plasma exchange in the management of acute immunoallergic thrombocytopenic purpura (author's transl)]. / Intérêt des échanges plasmatiques dans le traitement des purpuras thrombopéniques immuno-allergiques aigus.

Plasma exchanges were performed in 5 patients with acute immunoallergic thrombocytopenic purpura (ITP) because of severe haemorrhages and/or inefficient or badly tolerated corticosteroid therapy. Plasma exchanges were carried out with cell separators and were usually well tolerated. They resulted in very rapid increase in platelet levels with disappearance of auto-antibodies and dramatic reduction of haemorrhages, and they brought about, or accelerated, recovery in 3 patients. One patient was slightly improved. In the 5th patient, who had meningeal haemorrhage, plasma exchange was effective in increasing platelet levels but was unable to prevent a fatal outcome. The main indications for plasma exchange seems to be acute ITP with severe, life-threatening haemorrhages.