Studies on resistance to infectious pancreatic necrosis virus in Atlantic salmon (Salmo salar)

Infectious pancreatic necrosis virus (IPNV) is responsible for serious mortalities in postsmolt Atlantic salmon (Salmo salar) annually in both Scotland and Norway. The huge financial losses involved and the lack of control measures have prompted attempts to produce IPN vaccines and disease resistant strains of fish. A characteristic of IPNV is its ability to persist in a latent carrier state and to ensure stocks of fish are truly virus free there is a need for highly sensitive diagnostic methods. In this study microsatellites were employed to assign fish susceptible to IPN in the field to families. Laboratory challenges were performed on 24 families and various humoral immune parameters were measured. A variety of diagnostic techniques were also developed, their sensitivity and specificity evaluated and compared to tissue culture. Parental Atlantic salmon and offspring susceptible to IPN in the field were genotyped using a published multiplex microsatellite PCR adapted for use with fluorescent genotyping systems. The reliability and utility of the genotyping system was evaluated with positive results. The majority of susceptible fish could not be assigned to parents since the fish were found to be from a mixed stock. Various experimental challenges with IPNV infected tissue homogenate were performed on post-smolts, however they did not produce mortality or induce clinical IPN. As a result of this virus titre was used as an indirect measurement of disease resistance. Atlantic salmon post-smolts were found to effectively clear cell cultured IPNV, isolate Br from Shetland. A second Shetland isolate (Os) was found to actively replicate in the fish while isolate Br was rapidly cleared. To increase virulence isolate Os was passaged once further through fish prior to use in challenging 24 families of PIT 0 8 tagged Atlantic salmon. At week 3 mean viral titres for each family ranged from 10 to 1011 TCID5o/g kidney. Using these data six families were targeted to follow through to week 7, two with low titres, two with medium titres and two with high titres of IPNV. 0 8 15 At week 7 virus titres had decreased to between 10 ' and 1 0 '' TCIDso/g kidney, many of the fish in the families with low IPNV titres at week 3 had cleared the virus. Lysozyme and neutralising antibody responses were measured, however little or no correlation was found between either parameter and IPNV titre at weeks 3 or 7. The control fish were found to be positive for IPNV despite earlier negative testing. Reasons for this were unclear, the fish may have been exposed to the virus through the seawater inflow or alternatively the fish may have been carriers of the virus. Diagnostic techniques employed to detect IPNV were RT-PCR, ELISA and immunohistochemistry; cell culture was used as a comparison to the RT-PCR and ELISA. The RT-PCR was developed using two published IPNV primer sets both of which amplified regions on VP2, they were found to be specific to IPNV and together amplified 9 out of 10 serotypes of IPNV. The most sensitive primer set detected between 107 and 108 TCID50 IPNV/ml in spiked tissue, in comparison to the ELISA which detected 1012'5 TCID50 IPNV/ml in tissue homogenate. The AS-1 monoclonal antibody was used successfully in immunohistochemistry to identify IPNV in fixed tissue. The antibody was found to identify IPNV in pancreatic lesions from infected fish, but did not detect the virus in the gut. All of these methods were found to be less sensitive than cell culture, however they are very rapid and ELISA and RT-PCR may be used to confirm IPNV infection once CPE has been visualised in cell culture

Parents with psychosis : a pilot study examining self-report measures related to family functioning

The aims of this study were to examine the utility of various self-report instruments related to family functioning in families where a parent has a psychotic disorder, and to explore associations between these instruments and symptoms in the parent. Twenty-one parents with a psychotic disorders participated in the study. All participants were able to complete the questionnaires and the majority of parents reported levels of parental competence in the average range. Most parents (90%) perceived themselves to be effective parents, however 30% reported low levels of satisfaction with the parenting role. There were significant associations between objective measures of negative symptoms and self-report scores related to problems in ways of coping, and problems with parent-child interactions. Many individuals with psychosis were able to report areas of perceived need related to their role as parents and to the functioning of their families, however there are several limitations in the use of these instruments in this setting.<br /

The Anti-Apoptotic Effect of Respiratory Syncytial Virus on Human Peripheral Blood Neutrophils is Mediated by a Monocyte Derived Soluble Factor

Respiratory Syncytial Virus (RSV) causes annual epidemics of respiratory disease particularly affecting infants. The associated airway inflammation is characterized by an intense neutrophilia. This neutrophilic inflammation appears to be responsible for much of the pathology and symptoms. Previous work from our group had shown that there are factors within the airways of infants with RSV bronchiolitis that inhibit neutrophil apoptosis. This study was undertaken to determine if RSV can directly affect neutrophil survival

Long Term Displacement Data of Woven Fabric Webbings Under Constant Load for Inflatable Structures

Inflatable modules for space applications offer weight and launch volume savings relative to current metallic modules. Limited data exist on the creep behavior of the restraint layer of inflatable modules. Long-term displacement and strain data of two high strength woven fabric webbings, Kevlar and Vectran, under constant load is presented. The creep behavior of webbings is required by designers to help determine service life parameters of inflatable modules. Four groups of different webbings with different loads were defined for this study. Group 1 consisted of 4K Kevlar webbings loaded to 33% ultimate tensile strength and 6K Vectran webbings loaded to 27% ultimate tensile strength, group 2 consisted of 6K Kevlar webbings loaded to 40% and 43% ultimate tensile strength, and 6K Vectran webbings loaded to 50% ultimate tensile strength, group 3 consisted of 6K Kevlar webbings loaded to 52% ultimate tensile strength and 6K Vectran webbings loaded to 60% ultimate tensile strength, and group 4 consisted of 12.5K Kevlar webbings loaded to 22% ultimate tensile strength, and 12.5K Vectran webbings loaded to 22% ultimate tensile strength. The uniquely designed test facility, hardware, displacement measuring devices, and test data are presented. Test data indicate that immediately after loading all webbings stretch an inch or more, however as time increases displacement values significantly decrease to fall within a range of several hundredth of an inch over the remainder of test period. Webbings in group 1 exhibit near constant displacements and strains over a 17-month period. Data acquisition was suspended after the 17th month, however webbings continue to sustain load without any local webbing damage as of the 21st month of testing. Webbings in group 2 exhibit a combination of initial constant displacement and subsequent increases in displacement rates over a 16-month period. Webbings in group 3 exhibit steady increases in displacement rates leading to webbing failure over a 3-month period. Five of six webbings experienced local damage and subsequent failure in group 3. Data from group 4 indicates increasing webbing displacements over a 7-month period. All webbings in groups 1, 2, and 4 remain suspended without any local damage as of the writing of this paper. Variations in facility temperatures over test period seem to have had limited effect on long-term webbing displacement data

“That's cool, you’re a musician and you drink”: Exploring entertainers’ accounts of their unique workplace relationship with alcohol

This qualitative research investigates the alcohol experiences of entertainers who perform within licensed premises. Previous, mainly quantitative, studies have found that entertainers, specifically musicians, are an occupational group who drink excessively. This qualitative study draws on a wider sample of entertainers to examine their accounts of drinking in the workplace and the explanations they provide for this. We conducted individual semi-structured interviews (n = 24) with band-members, variety acts and DJs in Glasgow, Scotland. This revealed a workplace characterised by continual opportunities for often free alcohol consumption. Unlike most occupations, for entertainers ‘drinking-on-the-job’ was normative, expected, and sometimes encouraged by peers, the public, employers or sponsors. Entertainers also experienced performance-related incentives to drink before, during and/or after a show; including anxiety, matching their intoxication level to the audience's, and ‘reward-drinking’. This qualitative research confirms the unique nature of the entertainer-alcohol link, even in comparison to that found within other leisure industry occupations. While providing some explanation as to why entertainers might drink excessively, participants’ accounts also suggested potential strategies for avoiding the negative outcomes of workplace drinking

EDS1 complexes are not required for PRR responses and execute TNL‐ETI from the nucleus in Nicotiana benthamiana

Heterodimeric complexes incorporating the lipase-like proteins EDS1 with PAD4 or SAG101 are central hubs in plant innate immunity. EDS1 functions encompass signal relay from TIR domain-containing intracellular NLR-type immune receptors (TNLs) towards RPW8-type helper NLRs (RNLs) and, in Arabidopsis thaliana, bolstering of signaling and resistance mediated by cell-surface pattern recognition receptors (PRRs). Increasing evidence points to the activation of EDS1 complexes by small molecule binding. We used CRISPR/Cas-generated mutant lines and agroinfiltration-based complementation assays to interrogate functions of EDS1 complexes in Nicotiana benthamiana. We did not detect impaired PRR signaling in N. benthamiana lines deficient in EDS1 complexes or RNLs. Intriguingly, in assays monitoring functions of SlEDS1-NbEDS1 complexes in N. benthamiana, mutations within the SlEDS1 catalytic triad could abolish or enhance TNL immunity. Furthermore, nuclear EDS1 accumulation was sufficient for N. benthamiana TNL (Roq1) immunity. Reinforcing PRR signaling in Arabidopsis might be a derived function of the TNL/EDS1 immune sector. Although Solanaceae EDS1 functionally depends on catalytic triad residues in some contexts, our data do not support binding of a TNL-derived small molecule in the triad environment. Whether and how nuclear EDS1 activity connects to membrane pore-forming RNLs remains unknown

Reducing problem behavior during care-giving in families of preschool-aged children with developmental disabilities

This study evaluated two variants of a behavioral parent training program known as Stepping Stones Triple P (SSTP) using 74 preschool-aged children with developmental disabilities. Families were randomly allocated to an enhanced parent training intervention that combined parenting skills and care-giving coping skills (SSTP-E), standard parent training intervention alone (SSTP-S) or waitlist control (WL) condition. At post-intervention, both programs were associated with lower levels of observed negative child behavior, reductions in the number of care-giving settings where children displayed problem behavior, and improved parental competence and satisfaction in the parenting role as compared with the waitlist condition. Gains attained at post-intervention were maintained at 1-year follow-up. Both interventions produced significant reductions in child problem behavior, with 67% of children in the SSTP-E and 77% of children in the SSTPS showing clinically reliable change from pre-intervention to follow-up. Parents reported a high level of satisfaction with both interventions

Macrophages Are Required for Dendritic Cell Uptake of Respiratory Syncytial Virus from an Infected Epithelium

We have previously shown that the respiratory syncytial virus [RSV] can productively infect monocyte derived dendritic cells [MoDC] and remain dormant within the same cells for prolonged periods. It is therefore possible that infected dendritic cells act as a reservoir within the airways of individuals between annual epidemics. In the present study we explored the possibility that sub-epithelial DCs can be infected with RSV from differentiated bronchial epithelium and that in turn RSV from DCs can infect the epithelium. A dual co-culture model was established in which a differentiated primary airway epithelium on an Air Liquid Interface (ALI) was cultured on a transwell insert and MoDCs were subsequently added to the basolateral membrane of the insert. Further experiments were undertaken using a triple co-culture model in which in which macrophages were added to the apical surface of the differentiated epithelium. A modified RSV [rr-RSV] expressing a red fluorescent protein marker of replication was used to infect either the MoDCs or the differentiated epithelium and infection of the reciprocal cell type was assessed using confocal microscopy. Our data shows that primary epithelium became infected when rr-RSV infected MoDCs were introduced onto the basal surface of the transwell insert. MoDCs located beneath the epithelium did not become infected with virus from infected epithelial cells in the dual co-culture model. However when macrophages were present on the apical surface of the primary epithelium infection of the basal MoDCs occurred. Our data suggests that RSV infected dendritic cells readily transmit infection to epithelial cells even when they are located beneath the basal layer. However macrophages appear to be necessary for the transmission of infection from epithelial cells to basal dendritic cells

Validation of the GALS musculoskeletal screening exam for use in primary care: a pilot study

<p>Abstract</p> <p>Background</p> <p>As the proportion of the Canadian population ≥65 grows, so too does the prevalence of musculoskeletal (MSK) conditions. Approximately 20% of visits to family physicians occur as a result of MSK complaints. The GALS (Gait, Arms, Legs, and Spine) screening examination was developed to assist in the detection of MSK abnormalities. Although MSK exams are primarily performed by rheumatologists or other MSK specialists, expanding their use in primary health care may improve the detection of MSK conditions allowing for earlier treatment. The primary goal of this study was to evaluate the use of the GALS locomotor screen in primary care by comparing the results of assessments of family physicians with those of rheumatologists. The secondary goal was to examine the incidence of MSK disorders and assess the frequency with which new diagnoses not previously documented in patients' charts were identified.</p> <p>Methods</p> <p>Patients ≥65 years old recruited from an academic family health centre were examined by a rheumatologist and a family physician who recorded the appearance of each participant's gait and the appearance and movement of the arms, legs and spine by deeming them normal or abnormal. GALS scores were compared between physicians with the proportion of observed (P_obs), positive (P_pos) and negative (P_neg) agreement being the primary outcomes. Kappa statistics were also calculated. Descriptive statistics were used to describe the number of "new" diagnoses by comparing rheumatologists' findings with each patient's family practice chart.</p> <p>Results</p> <p>A total of 99 patients consented to participate (92 with previously diagnosed MSK conditions). Results showed reasonable agreement between family physicians and rheumatologists; P_obs= 0.698, P_pos= 0.614 and P_neg= 0.752. The coefficient of agreement (estimated Kappa) was 0.3675 for the composite GALS score. For individual components of the GALS exam, the highest agreement between family physicians and rheumatologists was in the assessment of gait and arm movement.</p> <p>Conclusion</p> <p>Previously reported increases in undiagnosed signs and symptoms of musculoskeletal conditions have highlighted the need for a simple yet sensitive screening exam for the identification of musculoskeletal abnormalities. Results of this study suggest that family physicians can efficiently use the GALS examination in the assessment of populations with a high proportion of musculoskeletal issues.</p

Drinking behaviours and blood alcohol concentration in four European drinking environments: a cross-sectional study

<p>Abstract</p> <p>Background</p> <p>Reducing harm in drinking environments is a growing priority for European alcohol policy yet few studies have explored nightlife drinking behaviours. This study examines alcohol consumption and blood alcohol concentration (BAC) in drinking environments in four European cities.</p> <p>Methods</p> <p>A short questionnaire was implemented among 838 drinkers aged 16-35 in drinking environments in four European cities, in the Netherlands, Slovenia, Spain and the UK. Questions included self-reported alcohol use before interview and expected consumption over the remainder of the night. Breathalyser tests were used to measured breath alcohol concentration (converted to BAC) at interview.</p> <p>Results</p> <p>Most participants in the Dutch (56.2%), Spanish (59.6%) and British (61.4%) samples had preloaded (cf Slovenia 34.8%). In those drinking < 3 h at interview, there were no differences in BAC by gender or nationality. In UK participants, BAC increased significantly in those who had been drinking longer, reaching 0.13% (median) in females and 0.17% in males drinking > 5 h. In other nationalities, BAC increases were less pronounced or absent. High BAC (> 0.08%) was associated with being male, aged > 19, British and having consumed spirits. In all cities most participants intended to drink enough alcohol to constitute binge drinking.</p> <p>Conclusions</p> <p>Different models of drinking behaviour are seen in different nightlife settings. Here, the UK sample was typified by continued increases in inebriation compared with steady, more moderate intoxication elsewhere. With the former being associated with higher health risks, European alcohol policy must work to deter this form of nightlife.</p