Susceptibility of pike Esox lucius to VHSV and IHNV and potential transmission to rainbow trout Oncorhynchus mykiss.

Determining the origin of recurrent outbreaks of fish diseases occurring on fish farms is essential for disease prevention and control measures. In this study, we investigated the potential reservoir role of wild fish species living near salmonid farms which were regularly found to be positive for viral hemorrhagic septicemia virus (VHSV). In addition to VHSV, infectious hematopoietic necrosis virus (IHNV) was also isolated from several pike Esox lucius samples collected from a pond near the salmonid farms of interest. All isolates of VHSV and IHNV analyzed had 100% identical partial glycoprotein gene sequences. VHSV pike strain OO128-25 belonged to the Ia genotype and shared 99.1 to 99.5% nucleotide identity with strains recently isolated from the farms. IHNV pike strain OO121-8, European genotype, appeared to be different from strains from France characterized since the first isolation in 1987. Isolates representative of both viral species were highly virulent in rainbow trout Oncorhynchus mykiss. OO128-25 induced 65% mortality in pike fingerlings, whereas only weak mortality was observed with OO121-8, despite characteristic symptoms in infected fish. High levels of specific antibodies to VHSV and IHNV were detected in adult pike in the absence of clinical signs. Infection of rainbow trout in contact with experimentally VHSV- or IHNV-infected pike fingerlings indicates possible horizontal transmission. These results suggest that pike could act as a reservoir for VHSV and IHNV in the wild, providing additional evidence to explain viral persistence and resurgence in certain areas.

First detection of Cyprinid Herpesvirus 2 (CyHV-2) in goldfish (Carassius auratus) in France.

Massive mortalities of Carassius auratus (L.) occurred in a farm in France during summer 2014. Fish presented anorexia, loss of scales and large amounts of mucus on the gills. Necrosis of the distal tip of the filament and the lamellae, combined with fusion of the lamellae, was observed, as well as necrosis in the hematopoietic organs and in the digestive tract. The histological examination led to hypothesize the implication of a virus in the mortality. The presence of cyprinid herpesvirus 2 (CyHV-2) in dead fish was demonstrated by amplification and sequencing of portions of the DNA polymerase and helicase genes, both sequences exhibiting 100% identity with CyHV-2 from Japan. In an attempt to find genetic markers of variation, two regions containing tandem repeats in the Japanese genome were amplified from a virus-positive sample from the present outbreak. A first region (mB) was fully identical to the Japanese isolate. However, the second region (mA) exhibited a range of deletions and substitutions compared to CyHV-2 from Japan. This is the first report of CyHV-2 in France in association with mortality of goldfish and the first identification of a molecular marker for its tracing.

Fishpathogens.eu/noda: a free and handy online platform for Betanodavirus targeted research and data sharing.

Viral nervous necrosis (VNN) is a severe neuropathological disease affecting a broad variety of finfish species worldwide. The causative agents of VNN are small viruses with a bi-segmented RNA genome known as betanodaviruses. At least four species with distinct but yet insufficiently characterized epidemiological features are recognized. The spread of VNN to an increasing number of host species, its wide geographic extent and its economical and ecological impacts justify the importance of collating as much molecular data as possible for tracing the origin of viral isolates and highlight the need for a freely accessible tool for epidemiological and molecular data sharing and consultation. For this purpose, we established a web-based specific database using the www.fishpathogens.eu platform, with the aim of collecting molecular and epidemiological information on VNN viruses, with relevance to their control, management and research studies.

First isolation of hirame rhabdovirus from freshwater fish in Europe.

A rhabdovirus was isolated in cell culture inoculated with tissue material from diseased grayling, Thymallus thymallus (L.), originating from a fish farm affected by a mortality episode in Poland. Diagnostics tests showed that the virus was not related to novirhabdoviruses known in Europe, nor to vesiculovirus-like species, except perch rhabdovirus (PRhV) with which it shared moderate serological relations. However, RT-PCR with PRhV probes gave negative results. To identify the virus, a random-priming sequence-independent single primer amplification was adopted. Surprisingly, two of the obtained sequences exhibited a high identity (>99%) with hirame rhabdovirus (HIRRV), a novirhabdovirus usually found in fish in marine Asiatic countries, for instance Japan, China and Korea. The full-length sequence of the phosphoprotein gene (P) demonstrated a higher identity of the present isolate with HIRRV from China compared with the Korean isolate. An identical viral sequence was also found in brown trout, Salmo trutta trutta L., affected by mortalities in a second farm in the same region, after a likely contamination from the grayling farm. To our knowledge, this is the first report of HIRRV in Europe, and in two hosts from fresh water that have not been described before as susceptible species.

[Awareness disorders in Alzheimer's disease and in mild cognitive impairment]. / Conscience des troubles dans la maladie d'Alzheimer et le mild cognitive impairment.

INTRODUCTION: Awareness disorders in Alzheimer's disease still remains unclear despite much research regarding this phenomenon. Papers report various and contrasted results with varying frequency from one study to another. Hence, the interest in awareness in Alzheimer's disease remains limited. Nevertheless, this symptom is closely associated with caregivers' burden and increases the patient's dependency, since the patient is unable to avoid dangers, requiring some care services or institutionalization The purpose of this current review is to recall the main neuro-anatomical and theoretical basis of awareness disorders, and to highlight the recent findings in Alzheimer's disease and in its pre-clinical stages. METHOD: With this in mind, we have conducted a non-exhaustive search using the pubmed online database to collect the most important reviews and the most recent findings regarding awareness disorders in Mild Cognitive Impairment (MCI) and/or in Alzheimer's disease. LITERATURE FINDINGS: In Alzheimer's disease, the links between awareness disorders and other variables, such as severity of dementia or depression, change from one study to the other and do not permit one to understand whether unawareness is an intrinsic or extrinsic reaction to the pathological process itself. Recent results suggest executive, cognitive and behavioral correlates more than psychopathological correlates, although the latter cannot be excluded. In Mild Cognitive Impairment, studies show varied results. Some studies report that patients suffering from Mild Cognitive Impairment can be compared to healthy control subjects and both groups have better awareness than patients with Alzheimer's disease. However, other studies show contrary results and awareness disorders might be a predictor of conversion from Mild Cognitive Impairment to dementia, as with apathy, in which the ability to cope with difficulties represents one of the main features. DISCUSSION: These controversial results are due to the heterogeneity of Alzheimer patients and in particular of MCI patients, but also to various conceptions of awareness disorders in Alzheimer's disease; none of them, however, taking into account all its diversity and complexity. Thus, neurological approaches underline neuropsychological dysfunctions linked to right frontal and/or hemispheric damage but are based on brain injury or strokes, which are events that differ greatly from a neurodegenerative disease involving progressive cognitive, emotional and social disturbances. Psychiatric approaches have taken into account the various aspects of insight, which before were often forbidden and reduced to a categorical point of view, and so could contribute to a better understanding of awareness disorders in Alzheimer's disease. However, these aspects have been conceptualized for psychiatric patients, suffering from positive symptoms, where compliance in treatment is the central key. Insight in neurological diseases is more focused on negative symptoms and generally concerns a basic perception of impairments in mainly cognitive domains. Moreover, modeling has often opposed neurological and psychological mechanisms, so awareness disorders are out of scope of a primary and secondary symptomatology. Although some authors have proposed to take into account these two mechanisms (unawareness and denial), clinical practice has shown that it was impossible to distinguish them both. Finally, some social cognitive approaches are able to demonstrate that there is no correlation between severity of dementia and awareness disorders. Nevertheless, the Self-concept, underlying this point of view, does not permit distinguishing neuropsychological from psycho-social factors. Moreover, only one evaluation tool based on Self-modeling in Alzheimer's disease exists, and to conceptualize Self in a specific pathology does not permit the comparison of this pathology to others or to healthy control subjects. So, the authors present the multidimensional model proposed by Clare et al., and some perspectives to stimulate future research, and perform potential meta-analyses.

Potential of Marine Strains of Pseudoalteromonas to Improve Resistance of Juvenile Sea Bass to Pathogens and Limit Biofilm Development.

The European sea bass (Dicentrarchus labrax), one of the most produced marine fish species in Europe, is acutely vulnerable to multiple infectious hazards. In this study, we investigated the potential probiotic effect of some marine Pseudoalteromonas bacterial strains against two major pathogens of this species, Vibrio harveyi and the nervous necrosis virus (NNV), and examined their antibiofilm effect. Impregnation phase was done by repeated immersion of juvenile's sea bass during 8 to 12 weeks in seawater containing the probiotic candidates at a concentration of 106 CFU/mL. Four candidates were tested: (1) a combination of two strains producing antimicrobial compounds, hCg-42 and hOe-125; (2) strain 3J6, with known antibiofilm properties; (3) strain RA15, from the same genus, but with no identified probiotic effect; and (4) a control group without probiotics. At the end of the impregnation phase, fish underwent an infection challenge with V. harveyi or with a pathogenic strain of NNV and mortality was monitored. For the V. harveyi challenge, improved survival rates of 10 and 25% were obtained for the RA15 and the mix hCg-42 + hOe-125-impregnated groups, respectively. For the NNV challenge, no significant benefic effect of the probiotics on infection kinetics or cumulative mortality was observed. At the end of the impregnation phase, the maximal thickness of biofilm was significantly lower in the 3J6, double strain, and RA15 groups, compared with the non-impregnated control group. This study highlights the interesting probiotic potential of marine bacteria to limit mortalities induced by bacterial pathogens as well as biofilm development.

[Tracheal lipoma: An exceptional cause of tracheal tumors]. / Lipome trachéal : une cause exceptionnelle de tumeur trachéale.

INTRODUCTION: Tracheal tumors are rare, they are most often malignant and can manifest themselves by a non-specific respiratory symptomatology with progressively increasing dyspnea orienting in the first place towards a COPD or even an asthma. Among them, tracheal lipoma is exceptional. Its management is based on removal by rigid bronchoscopy. OBSERVATION: We report the case of a 73-year-old male patient who presented with non-specific dyspnea that progressively worsened over several months. The EFR showed a flattening of the flow-volume curves, the CT scan showed an anterolateral oval tracheal tumor with fatty density, the bronchial endoscopy showed a tumor lesion with stenosis of about 90% of the airway. Management consisted of a rigid bronchoscopy to delete obtruction with biopsies. Anatomopathology concluded to a tracheal lipoma. CONCLUSION: Progressively worsening dyspnea, especially if there are signs of inspiratory dyspnea, required a systematic bronchial endoscopy to avoid the possibility of a tracheal tumor.

Comparing residential programmes for adults with autism spectrum disorders and intellectual disability: outcomes of challenging behaviour and quality of life.

BACKGROUND: Owing to methodological issues, little research has been conducted to examine quality of life (QoL) as a treatment outcome in autism spectrum disorders (ASD) and intellectual disabilities (ID). This study was conducted to combine QoL measures and objective observations of challenging behaviours (CB) in order to evaluate changes over time in adults with ASD and ID who were treated in two different residential programmes; we hypothesised that a decrease in CB would be related to an improved QoL. METHOD: In a longitudinal study (45 months), we followed 31 adults with ASD and ID who had been integrated into two residential programmes [Autism Programme with a Structured Method (PAMS) vs. traditional programme for ID (No-PAMS)] for 2-19 years. QoL [Quality of Life Inventory in a Residential Environment (IQVMR)] and severity of autistic features (Childhood Autism Rating Scales) were evaluated annually. CB, as measured by the Aberrant Behaviour Checklist (ABC), including stereotypic behaviour and inappropriate speech, were repeatedly assessed every 3 months. RESULTS: Observed separately, the groups' results were different. In the PAMS programme, stereotypic behaviour and inappropriate speech (ABC scores) significantly decreased, and the IQVMR total score increased; in contrast, in the comparison group, ABC scores did not change and the IQVMR total score decreased. In all, three mixed-effect ANCOVAs partially confirmed that the PAMS programme had an effect on CB and that QoL improvement did not directly depend on the type of programme but on reducing CB as measured by the ABC. CONCLUSION: The PAMS programme has a positive and indirect influence on QoL by reducing CB.

[Rapid outpatient diagnostic pathways for lung cancer: Evaluation after one year]. / Parcours de diagnostic rapide du cancer du poumon : évaluation à un an.

INTRODUCTION: The introduction of coordinated care pathways for lung cancer diagnosis and treatment is a complex process. The purpose of the French Cancer Plan 2014-2019 was to improve referral to treatment waiting times in people with suspected malignancy. The aim of this study was to assess a rapid outpatient diagnostic program for lung cancer established in 2016. METHOD: This retrospective study was carried out in the Pulmonology Department at Tenon Hospital, Paris, France between May 2016 and May 2017. RESULTS: During this period, 118 patients (60%) of patients in the pathway were diagnosed with lung cancer. The median waiting time to first consultation (D1) was 4 (2-7) days. The median waiting time between diagnosis and treatment decision (D4) was 4 (0-8) days. The median waiting time to the first treatment (D5) was 10 (4-15) days for chemotherapy and 27 (16-34) days for surgery. The median waiting time between the first abnormal chest X-ray and the first treatment (D6) was 49 days (34-70). CONCLUSION: Referral to treatment waiting times was consistent with international recommendations. Coordinating nurses improved care pathways in lung cancer patients.

Correlation between the Epworth Sleepiness Scale and the Maintenance of Wakefulness Test in Obstructive Sleep Apnea Patients Treated with Positive Airway Pressure.

BACKGROUND: The Epworth sleepiness scale (ESS) is often used to evaluate the impact of treatment in patients with obstructive sleep apnea hypopnea syndrome (OSA). We aimed to evaluate the correlation between ESS and the Maintenance of Wakefulness Test (MWT) in a population of OSA patients treated with positive airway pressure (PAP). METHODS: We retrospectively included all patients during a 2-year period who were diagnosed with OSA in our sleep clinic and required PAP therapy. ESS was evaluated at baseline and after PAP therapy for all patients, and all had a concomitant MWT. Correlation between final ESS, change in ESS, and MWT were evaluated using Spearman's correlation. Given that MWT is considered as the gold standard, the diagnostic performance of ESS was evaluated against MWT. RESULTS: Hundred thirty-four OSA patients were included. At the time of MWT, 89.6% of the patients were compliant (PAP use ≥4hours/night), and only 9 (6.7%) had persistent sleepiness despite PAP treatment (mean sleep latency at MWT<19.4min). Moderate correlation was observed between final ESS and MWT (Spearman's correlation coefficient=-0.42), but no correlation was found between change in ESS and MWT. Diagnostic performance was as follows for final ESS: sensitivity=55.6%, specificity=84.8%, PPV=20.8%, and NPV=96.4%. CONCLUSIONS: ESS was moderately correlated with MWT in a population of OSA patients compliant with PAP therapy. In this population, ESS showed poor diagnostic performance in identifying patients with persistent excessive daytime sleepiness. CLINICALTRIALS. GOV IDENTIFIER: NCT03629834.

Outbreak of betanodavirus infection in tilapia, Oreochromis niloticus (L.), in fresh water.

A betanodavirus associated with a massive mortality was isolated from larvae of tilapia, Oreochromis niloticus, maintained in fresh water at 30 degrees C. Histopathology revealed vacuolation of the nervous system, suggesting an infection by a betanodavirus. The virus was identified by indirect fluorescent antibody test in the SSN1 cell line and further characterized by sequencing of a PCR product. Sequencing of the T4 region of the coat protein gene indicated a phylogenetic clustering of this isolate within the red-spotted grouper nervous necrosis virus type. However, the tilapia isolate formed a unique branch distinct from other betanodavirus isolates. The disease was experimentally reproduced by bath infection of young tilapia at 30 degrees C. The reservoir of virus at the origin of the outbreak remains unidentified. To our knowledge, this is the first report of natural nodavirus infection in tilapia reared in fresh water.

Quality of life of adults with pervasive developmental disorders and intellectual disabilities.

The purpose of this study was to observe quality of life (QoL) and global evolution of persons with Pervasive Developmental Disorders (PDD) in three different groups. Individualized programs for PDD were compared to traditional programs for intellectual disabilities. Behavioural disorders were repeatedly evaluated using the Aberrant Behaviour Checklist (ABC) and QoL once a year. Little research has investigated this domain due to methodological problems with a non-verbal population. Two preliminary studies of individualized programs showed a significant reduction in behaviour disorders over the course of the study. The recent inclusion of a control group indicates that a traditional program reduces lethargy/social withdrawal (ABC factor 2). A good QoL was measured for the three groups.

Optimization of a "bump-and-hole" approach to allele-selective BET bromodomain inhibition.

Allele-specific chemical genetics enables selective inhibition within families of highly-conserved proteins. The four BET (bromodomain & extra-terminal domain) proteins - BRD2, BRD3, BRD4 and BRDT bind acetylated chromatin via their bromodomains and regulate processes such as cell proliferation and inflammation. BET bromodomains are of particular interest, as they are attractive therapeutic targets but existing inhibitors are pan-selective. We previously established a bump-&-hole system for the BET bromodomains, pairing a leucine/alanine mutation with an ethyl-derived analogue of an established benzodiazepine scaffold. Here we optimize upon this system with the introduction of a more conservative and less disruptive leucine/valine mutation. Extensive structure-activity-relationships of diverse benzodiazepine analogues guided the development of potent, mutant-selective inhibitors with desirable physiochemical properties. The active enantiomer of our best compound - 9-ME-1 - shows ∼200 nM potency, >100-fold selectivity for the L/V mutant over wild-type and excellent DMPK properties. Through a variety of in vitro and cellular assays we validate the capabilities of our optimized system, and then utilize it to compare the relative importance of the first and second bromodomains to chromatin binding. These experiments confirm the primacy of the first bromodomain in all BET proteins, but also significant variation in the importance of the second bromodomain. We also show that, despite having a minor role in chromatin recognition, BRD4 BD2 is still essential for gene expression, likely through the recruitment of non-histone proteins. The disclosed inhibitor:mutant pair provides a powerful tool for future cellular and in vivo target validation studies.

[A prospective study of quality of life and treatment of chemotherapy-induced anaemia in lung cancer]. / Qualité de vie et traitement de l'anémie chimio-induite dans le cancer bronchopulmonaire: étude prospective.

INTRODUCTION: The impact of chemotherapy-induced anemia on the quality of life (QOL) of patients with lung cancer has been little studied. OBJECTIVE: We evaluated the feasibility of measuring QOL among patients receiving chemotherapy for lung cancer, and the possible correlation of QOL with the haemoglobin level. METHODS: This was a prospective study of 53 patients starting chemotherapy (total 155 cycles); QOL was measured with the specific Fact-An scale. RESULTS: The mean haemoglobin level before treatment was 13.1+/-2.3 g/dl. During chemotherapy 45.3% of patients received erythrocytic growth factors and 15.1% were transfused; 26.4% of patients refused to answer or could not answer the QOL questionnaire. There was a strong correlation between the Hb level and overall QOL (r=0.343, p=0.0004), as well as the physical and functional subscales (but not the cognitive and social subscales). CONCLUSIONS: Although QOL could not be measured in one-quarter of cases, it was clearly affected by anaemia, which supports a strategy of early diagnosis and management of anaemia due to chemotherapy for lung cancer.

Inhibition by two diphosphonates of bone lysis in tumor-conditioned media.

In order to test the effect of diphosphonates in inhibiting bone lysis induced by human tumors, neonatal mouse calvaria are cultured in sterile conditions in different media. Osteolysis is estimated by the amount of 45Ca released from bone to medium, the mice being given injections of 45Ca on their day of birth. An increased bone lysis is observed when calvaria are incubated in the same medium conditioned from cultured tumor fragments. This effect is significantly decreased when the mice have been treated with either ethanehydroxydiphosphonate or dichloromethylenediphosphonate. These experiments represent a first step in estimating the potential effects of different drugs on malignant osteolysis.

First generic one step real-time Taqman RT-PCR targeting the RNA1 of betanodaviruses.

The detection of betanodavirus genomic components is a major issue for diagnostics and control of viral nervous necrosis (VNN), a devastating disease affecting fish worldwide. Despite a number of published molecular-based tests, most of them targeting the RNA2 molecule of the virus, diagnostics is still a challenge due to the high genetic diversity within this genus. In the present study, a new one-step real-time RT-PCR (rRT-PCR), targeting RNA1 of most genotypes of betanodaviruses, was proposed and validated. The test detected successfully various isolates of betanodavirus representatives of the four species RGNNV, SJNNV, TPNNV and BFNNV, either produced on cell culture or from clinical samples. It was specific as shown by the absence of signal on samples from healthy sea bass or from field samples of six other fish species without clinical signs of VNN. The assay detected reliably 50-100 copies of plasmids containing the targeted cloned RNA1 region, as well as an infectious dose of virus of 10(2.5)-10(2.85) TCID50/ml. A set of samples was tested by two different laboratories, with similar results, demonstrating the robustness of the test. This is the first one step generic rRT-PCR method for betanodaviruses. It is simple to perform and may be used for first intention diagnostics as well as for confirmation in case of doubtful results obtained with other published tests targeting RNA2.

Effects of doxapram on hypercapnic response during weaning from mechanical ventilation in COPD patients.

Failure of weaning from mechanical ventilation in COPD patients is often related to diaphragmatic fatigue. Whether there is a central respiratory drive fatigue and a reserve of excitability is still debated. The purpose of this study was to analyze the following in 13 COPD patients weaned from mechanical ventilation: (1) ventilatory (VE/PETCO2) and neuromuscular (P0.1/PETCO2) response to hypercapnia; (2) the maximum reserve capacity measured through changes in the VE/PETCO2 and P0.1/PETCO2 slopes after doxapram (DXP) infusion, which, given during the test, allows measurement of the maximum response capacity to overstimulation; and (3) analyze the influence of these changes on the outcome of weaning. The results show a variable P0.1/PETCO2 response and a low VE/PETCO2. DXP infusion does not change the slopes of these relations but increases the end-expiratory volume (delta FRCd); (p less than 0.02). Since there was no change in the VE/PETCO2, P0.1/PETCO2, and delta FRC values with or without DXP, there was no excitability reserve in patients who were successfully weaned. When weaning failed, DXP did not change VE/PETCO2 and P0.1/PETCO2 slope, but delta FRCd was greater the delta FRC (p less than 0.001). The excitability reserve in these patients leads to an increase in end-expiratory volume, probably worsening the diaphragm dysfunction.