Cellular prion protein (PrPC) in the development of Merlin-deficient tumours.

Loss of function mutations in the neurofibromatosis Type 2 (NF2) gene, coding for a tumour suppressor, Merlin, cause multiple tumours of the nervous system such as schwannomas, meningiomas and ependymomas. These tumours may occur sporadically or as part of the hereditary condition neurofibromatosis Type 2 (NF2). Current treatment is confined to (radio) surgery and no targeted drug therapies exist. NF2 mutations and/or Merlin inactivation are also seen in other cancers including some mesothelioma, breast cancer, colorectal carcinoma, melanoma and glioblastoma. To study the relationship between Merlin deficiency and tumourigenesis, we have developed an in vitro model comprising human primary schwannoma cells, the most common Merlin-deficient tumour and the hallmark for NF2. Using this model, we show increased expression of cellular prion protein (PrPC) in schwannoma cells and tissues. In addition, a strong overexpression of PrPC is observed in human Merlin-deficient mesothelioma cell line TRA and in human Merlin-deficient meningiomas. PrPC contributes to increased proliferation, cell-matrix adhesion and survival in schwannoma cells acting via 37/67 kDa non-integrin laminin receptor (LR/37/67 kDa) and downstream ERK1/2, PI3K/AKT and FAK signalling pathways. PrPC protein is also strongly released from schwannoma cells via exosomes and as a free peptide suggesting that it may act in an autocrine and/or paracrine manner. We suggest that PrPC and its interactor, LR/37/67 kDa, could be potential therapeutic targets for schwannomas and other Merlin-deficient tumours.

The fine needle aspiration of translocation sarcomas.

INTRODUCTION: Soft tissue sarcomas comprise a heterogeneous group of clinically aggressive cancers that are often hard to classify on limited cytological samples. "Translocation sarcomas" (TS) are a diverse subset of such cancers, different from pleomorphic sarcomas, and characterised by unique single chromosomal translocations in each sarcoma subtype. Interestingly, despite their high-grade biological behaviour, TS have deceptively monotonous and bland cytomorphology, therefore creating diagnostic issues on limited samples. MATERIALS AND METHODS: A retrospective search was conducted of the cytopathology archives of The Johns Hopkins Hospital revealing 147 translocation sarcoma cases over a 25-year period. RESULTS: The common morphological denominators for most translocation sarcomas were: hypercellularity, cellular monotony, mostly discohesive and single cells, round-to-oval or short spindled cells and a lack of necrosis. The exceptions were an inflammatory myofibroblastic tumour, in which cellular monotony was not present owing to the prominence of lymphocytes and plasma cells, and low-grade fibromyxoid sarcoma, in which the specimens were generally hypocellular. Ancillary testing, especially immunoperoxidase staining, was often required for primary lesions. CONCLUSION: Distinct morphological clues and subsequent ancillary testing (particularly immunoperoxidase staining) provide an accurate diagnosis on cytological interpretation of both, primary and recurrent/metastatic lesions.

Dairy farmers' perceptions toward the implementation of on-farm Johne's disease prevention and control strategies.

Implementation of specific management strategies on dairy farms is currently the most effective way to reduce the prevalence of Johne's disease (JD), an infectious chronic enteritis of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP). However, dairy farmers often fail to implement recommended strategies. The objective of this study was to assess perceptions of farmers participating in a JD prevention and control program toward recommended practices, and explore factors that influence whether or not a farmer adopts risk-reducing measures for MAP transmission. Semi-structured interviews were conducted with 25 dairy farmers enrolled in a voluntary JD control program in Alberta, Canada. Principles of classical grounded theory were used for participant selection, interviewing, and data analysis. Additionally, demographic data and MAP infection status were collected and analyzed using quantitative questionnaires and the JD control program database. Farmers' perceptions were distinguished according to 2 main categories: first, their belief in the importance of JD, and second, their belief in recommended JD prevention and control strategies. Based on these categories, farmers were classified into 4 groups: proactivists, disillusionists, deniers, and unconcerned. The first 2 groups believed in the importance of JD, and proactivists and unconcerned believed in proposed JD prevention and control measures. Groups that regarded JD as important had better knowledge about best strategies to reduce MAP transmission and had more JD risk assessments conducted on their farm. Although not quantified, it also appeared that these groups had more JD prevention and control practices in place. However, often JD was not perceived as a problem in the herd and generally farmers did not regard JD control as a "hot topic" in communications with their herd veterinarian and other farmers. Recommendations regarding how to communicate with farmers and motivate various groups of farmers according to their specific perceptions were provided to optimize adoption of JD prevention and control measures and thereby increase success of voluntary JD control programs.

Short communication: Herd-level prevalence of Mycobacterium avium ssp. paratuberculosis is not associated with participation in a voluntary Alberta Johne's disease control program.

Johne's disease (JD) control programs for dairy farms have the general objective of reducing both cow- and herd-level prevalence of Mycobacterium avium ssp. paratuberculosis (MAP). An important aspect of many programs is herd testing for MAP to determine the infection status of participating farms. However, it is uncertain whether MAP herd-level prevalence on farms voluntarily participating in a JD control program is different from that on nonparticipating farms. Therefore, the aim was to compare MAP infection status of participants and nonparticipants in the Alberta Johne's Disease Initiative (AJDI), a voluntary JD control program initiated in 2010 in Alberta, Canada. Between September 2012 and August 2013, environmental fecal samples were collected from 93 randomly selected farms not enrolled in the AJDI. Additionally, 81 farms that initially enrolled in the AJDI during the same time interval were also sampled. Samples were collected from 6 defined locations on each farm and cultured for MAP. Results were confirmed using conventional IS900 PCR and F 0285 quantitative PCR. Overall, 51% of participating and 51% of nonparticipating farms were identified as being MAP-infected. Furthermore, based on multivariable logistic regression, the number of MAP-positive samples was not associated with AJDI participation (taking herd size into account as a potentially modifying or confounding variable). In conclusion, there was no indication that voluntary participation in the AJDI was associated with herd-level MAP prevalence.

Factors associated with participation of Alberta dairy farmers in a voluntary, management-based Johne's disease control program.

The Alberta Johne's Disease Initiative (AJDI) is a voluntary, management-based prevention and control program for Johne's disease (JD), a wasting disease in ruminants that causes substantial economic losses to the cattle industry. Despite extensive communication about the program's benefits and low cost to participating producers, approximately 35% of Alberta dairy farmers have not enrolled in the AJDI. Therefore, the objective was to identify differences between AJDI nonparticipants and participants that may influence enrollment. Standardized questionnaires were conducted in person on 163 farms not participating and 61 farms participating in the AJDI. Data collected included demographic characteristics, internal factors (e.g., attitudes and beliefs of the farmer toward JD and the AJDI), external factors (e.g., farmers' JD knowledge and on-farm goals and constraints), as well as farmers' use and influence of various information sources. Nonparticipants and participants differed in at least some aspects of all studied categories. Based on logistic regression, participating farms had larger herds, higher self-assessed knowledge of JD, better understanding of AJDI details before participation, and used their veterinarian more often to get information about new management practices and technologies when compared with nonparticipants. In contrast, nonparticipants indicated that time was a major on-farm constraint and that participation in the AJDI would take too much time. They also indicated that they preferred to wait and see how the program worked on other farms before they participated.

Standardised clients as assessors in a veterinary communication OSCE: a reliability and validity study.

In human medicine, standardised patients (SP) have been shown to reliably and accurately assess learners' communication performance in high-stakes certification Objective Structured Clinical Examinations (OSCE), offering a feasible way to reduce the need for recruitment, time commitment and coordination of faculty assessors. In this study, we evaluated the use of standardised clients (SC) as a viable option for assessing veterinary students' communication performance. We designed a four-station, two-track communication skills OSCE. SC assessors used an adapted nine-item Liverpool Undergraduate Communication Assessment Scale (LUCAS). Faculty used a 21-item checklist derived from the Calgary-Cambridge Guide (CCG) and a five-point global rating scale. Participants were second year veterinary students (n=96). For the four stations, intrastation reliability (α) ranged from 0.63 to 0.82 for the LUCAS, and 0.73 to 0.87 for the CCG. The interstation reliability coefficients were 0.85 for the LUCAS and 0.89 for the CGG. The calculated Generalisability (G) coefficients were 0.62 for the LUCAS and 0.60 for the CGG. Supporting construct validity, SC and faculty assessors showed a significant correlation between the LUCAS and CCG total percent scores (r=0.45, P<0.001), and likewise between the LUCAS and global rating scores (r=0.49, P<0.001).Study results support that SC assessors offer a reliable and valid approach for assessing veterinary communication OSCE.

Dog-walking and sense of community in neighborhoods: implications for promoting regular physical activity in adults 50 years and older.

This study investigates whether dog-ownership and neighborhood characteristics are associated with sense of community (SC) and neighborhood-based recreational walking (NRW) for older adults. A random sample of adults ≥50 years of age (n=884) provided information on SC, dog-related factors, neighborhood walking, and socio-demographics in telephone and postal surveys. Associations between dog-ownership, neighborhood characteristics, and NRW were estimated using logistic regression (i.e., odds ratios (OR)). Frequent dog-walkers (≥4 times/wk) were more likely than those not owning a dog to report a heightened SC (OR=1.94, p<.05) and to achieve ≥150min/wk of NRW (OR=10.68, p<.05). SC was also tested but not found to mediate associations between neighborhood characteristics, dog-ownership and NRW. Older adults who walk dogs often in their neighborhoods may benefit from both increased physical activity and heightened sense of community to an extent that supports healthy aging. Longitudinal studies are needed to explore directions of associations among these factors.

Vibrational autodetachment-intramolecular vibrational relaxation translated into electronic motion.

If a negative ion has vibrational energy in excess of the binding energy of its most weakly bound electron, the anion can undergo vibrational autodetachment, similar to thermionic emission. When this effect occurs after targeted infrared excitation of a specific vibrational mode in the anion, it encodes information on the intramolecular vibrational relaxation processes that take place between excitation and electron emission. We present examples on how vibrational autodetachment can be used to obtain infrared spectra of molecular anions, and we discuss how a vibrational autodetachment photoelectron spectrum can be modeled, using vibrational autodetachment after excitation of CH stretching modes of nitromethane anions as a case study.

Cue-conditioned alcohol seeking in rats following abstinence: involvement of metabotropic glutamate 5 receptors.

BACKGROUND AND PURPOSE: The current study was designed to: (i) examine whether functional interactions occur between receptors known to regulate alcohol self-administration; and (ii) characterize relapse to alcohol seeking following abstinence. EXPERIMENTAL APPROACH: The selective cannabinoid CB(1) receptor antagonist SR141716A (0.03-1.0 mg.kg(-1) i.p.) resulted in a dose-dependent reduction in ethanol self-administration in ethanol-preferring Indiana-preferring rats. SR141716A was then co-administered with either the selective glutamate metabotropic glutamate 5 (mGlu(5)) receptor antagonist 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (MTEP) or the selective adenosine A(2A) receptor antagonist SCH58261. KEY RESULTS: When administered at individually sub-threshold doses, a combination of SR141716A (0.1 mg.kg(-1)) and SCH58261 (0.5 mg.kg(-1) i.p.) produced a reduction (28%) in ethanol self-administration. Combinations of threshold doses of SR141716A (0.3 mg.kg(-1)) and SCH58261 (2.0 mg.kg(-1), i.p.) caused an essentially additive reduction (68%) in alcohol self-administration. A combination of individually sub-threshold doses of CB(1) and mGlu(5) receptor antagonists did not affect alcohol self-administration; however, combined threshold doses of SR141716A (0.3 mg.kg(-1)) and MTEP (1.0 mg.kg(-1) i.p.) did reduce ethanol self-administration markedly (80%). Cue-conditioned alcohol seeking was attenuated by pretreatment with MTEP (1.0 mg.kg(-1)) co-administered with SR141716A (0.3 mg.kg(-1) i.p.). In contrast, SCH58261 (2.0 mg.kg(-1)) co-administered with SR141716A (0.3 mg.kg(-1) i.p.) did not reduce cue-conditioned alcohol seeking. CONCLUSIONS AND IMPLICATIONS: Adenosine A(2A) and cannabinoid CB(1) receptors regulated alcohol self-administration additively, but combined low-dose antagonism of these receptors did not prevent cue-conditioned alcohol seeking after abstinence. In contrast, combined low-dose antagonism of mGlu(5) and CB(1) receptors did prevent relapse-like alcohol seeking after abstinence, suggesting a prominent role for mGlu(5) receptors in this paradigm.

Development and validation of an instrument for measuring appointment-specific client satisfaction in companion-animal practice.

Healthcare research recognizes that 'satisfaction' is an important health outcome of the medical encounter. As a result, many healthcare professions have pursued the development and validation of instruments for measuring patient satisfaction. However, veterinary medicine has developed and properly validated few instruments for measuring client satisfaction. This paper describes the development and psychometric assessment of an instrument for measuring appointment-specific client satisfaction in companion-animal practice. We developed the Client Satisfaction Questionnaire (CSQ) in four main phases. Phase I relied on a series of six pet-owner focus groups (n=32) to explore clients' expectations of veterinary care to provide a basis for constructing items for the CSQ. Phase II involved developing items for a first draft of the CSQ, using the content and themes taken from the pet-owner focus groups. In phase III we pre-tested the first draft with six veterinarians in companion-animal practice and a convenience sample of their clients (n=129). Data from the pre-test were used to test the initial psychometric characteristics of each item. Together with participant feedback, these findings were used to design the final, 15-item CSQ. Phase IV involved psychometrically testing the final, 15-item CSQ as part of a larger observational study involving 20 veterinarians and 344 of their clients. We used data from this sample to assess the reliability and validity of the CSQ in companion-animal practice. Assessment was based on descriptive statistics, principal-component analysis, generalizability theory and linear mixed modeling. Findings demonstrate excellent reliability (G-coefficient for internal consistency=0.96) and support the face, content and construct validity of the CSQ as a measure of appointment-specific client satisfaction in companion-animal practice.

Essential education in communication skills and cultural sensitivities for global public health in an evolving veterinary world.

In the practise of veterinary medicine and global public health, communication skill is as critical as clinical reasoning and an extensive knowledge base. Effective communication skills and cross-cultural sensitivity are essential across the board for interdisciplinary, international, and local veterinary medicine. This paper offers an evidence-based, three-part framework for developing and sustaining curricula that enhance communication skills and cross-cultural sensitivity so that students are better prepared to practise veterinary medicine in an evolving world. These curricula may well also serve as a conduit for encouraging more veterinary graduates to choose global public health as a career path.

The value of serum neopterin, interferon-gamma levels and interleukin-12B polymorphisms in predicting acute renal allograft rejection.

Acute rejection remains a poor predictor of graft outcome. In this study, we measured serum levels of interferon (IFN)-gamma and neopterin by enzyme-linked immunosorbent assay and a single nucleotide polymorphism (SNP) within the 3' untranslated region of the interleukin (IL)-12 B gene (1188 A/C) to determine whether either of these factors could predict acute rejection in renal transplantation. Significantly higher early post-transplant neopterin levels (days 5-7; 35.7 versus 19.9 nmol/l) were observed in recipients who subsequently rejected their grafts. Post-transplant neopterin levels showed a strong positive correlation with 1-month creatinine levels (Spearman's correlation 0.62, P < 0.001), suggesting macrophage activation early after transplantation. Pretransplant neopterin and IFN-gamma levels and the IL-12B gene SNP did not predict acute rejection in this small retrospective study. The ability to predict acute rejection non-invasively early after transplantation could lead to individual tailoring of immunosuppressive regimens and perhaps lead eventually to longer graft survival.

CGP7930: a positive allosteric modulator of the GABAB receptor.

CGP7930 (3-(3',5'-Di-tert-butyl-4'-hydroxy)phenyl-2,2-dimethylpropanol) is a positive allosteric modulator of the metabotropic GABAB receptor. CGP7930 has been found to modulate the GABAB receptor in the open, or high affinity, state increasing agonist affinity for the receptor and signal transduction efficacy following agonist stimulation. The GABAB heteromeric subunit B2, involved in signal transduction but not ligand binding, seems to be the site of action of CGP7930 and similar allosteric modulators. When administered alone in naïve animals, CGP7930 acts as an anxiolytic in rodents without other overt behavioral effects and has also been demonstrated to reduce self-administration of nicotine, cocaine, or alcohol in rodents, suggesting that "fine tuning" of the GABAB receptor by positive allosteric modulators may be able to regulate abuse of these drugs. Baclofen, the GABAB agonist, is currently finding use in treating addiction and various other disorders, but this can result in off-target effects and tolerance. CGP7930 when co-administered with baclofen enhances its potency, which could in theory minimize deleterious effects. Further study of CGP7930 is required, but this compound, and others like it, holds potential in a clinical setting.

Antibodies targeting the calcium binding skeletal muscle protein calsequestrin are specific markers of ophthalmopathy and sensitive indicators of ocular myopathy in patients with Graves' disease.

We have identified several eye muscle antigens and studied the significance of the corresponding serum autoantibodies in patients with Graves' disease. Of these antigens, only calsequestrin is expressed more in eye muscle than other skeletal muscles, which could explain at least partly the specific involvement of eye muscle in patients with Graves' disease. Earlier, we found a modest relationship between anti-calsequestrin antibodies and ophthalmopathy, but in that study we used calsequestrin prepared from rabbit heart muscle and measured antibodies by immunoblotting. We have reinvestigated the prevalences of anti-calsequestrin antibodies in larger groups of well-characterized patients with thyroid autoimmunity with and without ophthalmopathy and control patients and healthy subjects, using standard enzyme-linked immunosorbent assay incorporating highly purified rabbit skeletal muscle calsequestrin, which has a 97% homology with human calsequestrin, as antigen. Anti-calsequestrin antibodies were detected in 78% of patients with active congestive ophthalmopathy, in 92% of those with active inflammation and eye muscle involvement, but in only 22% of patients with chronic, 'burnt out' disease. Tests were also positive in 5% of patients with Graves' hyperthyroidism without evident ophthalmopathy (two patients) and one patient with 'watery eyes' but no other clear signs of congestive ophthalmopathy and IgA nephropathy and no known thyroid disease, but in no patient with Hashimoto's thyroiditis, toxic nodular goitre, non-toxic multi-nodular goitre or diabetes, or age- and sex-matched healthy subjects. In serial studies of all 11 patients with Graves' hyperthyroidism who had active ophthalmopathy at the time of the first clinic visit, or developed eye signs during the first 6 months, and positive anti-calsequestrin antibodies in at least one sample, anti-calsequestrin antibodies correlated with the onset of ocular myopathy in six patients. Antibodies targeting calsequestrin appear to be specific markers for ophthalmopathy and sensitive indicators of the ocular myopathy subtype of ophthalmopathy in patients with thyroid autoimmunity. However, these results must be considered preliminary until a large prospective study of patients with newly diagnosed Graves' hyperthyroidism, in which serum levels of calsequestrin antibodies are correlated with clinical changes and orbital eye muscle and connective tissue/fat volumes, has been carried out.

Survey of veterinary technical and professional skills in students and recent graduates of a veterinary college.

OBJECTIVES: To determine perceptions of veterinary technical and professional skills among veterinary students and recent graduates. DESIGN: Cross-sectional study. SAMPLE POPULATION: 281 students and 142 recent graduates from the Ontario Veterinary College. PROCEDURE: A survey was designed and administered to first- through fourth-year students and veterinarians who had graduated either 1 or 6 years before survey administration. RESULTS: Overall response rate was 70%. Learning about technical and professional skills was highly valued. Most participants felt they had not received instruction about professional skills, but those who had felt more competent about them. Perceptions of competence increased slightly with increased comfort discussing emotional veterinary issues with instructors. Neither gender nor increased age was related to increased feelings of competence. Almost all fourth-year students felt competent and comfortable about examining an animal with the client present, assessing suffering, diagnosing parvovirus infection, performing surgery, and working as group members. However, many did not feel competent or comfortable about delivering bad news, setting time limits yet providing quality service, helping clients with limited funds make treatment decisions, dealing with demanding people, and euthanasia. Feelings of competence and comfort were closely related but were not identical. CONCLUSIONS AND CLINICAL RELEVANCE: In the interests of best preparing entry-level veterinarians, technical and professional skills need to be emphasized in a learning environment where students feel comfortable discussing emotional veterinary issues. A professional skills curriculum addressing underlying self-awareness, communication, and interpersonal issues, as well as procedural matters, would likely increase the proportion of fourth-year students who feel competent and comfortable about professional skills by the end of their undergraduate training.

Predictors of owner response to companion animal death in 177 clients from 14 practices in Ontario.

OBJECTIVE: To identify predictors of grief and client desires and needs as they relate to pet death. DESIGN: Cross-sectional mail survey. SAMPLE POPULATION: 177 clients, from 14 randomly selected veterinary practices, whose cat or dog died between 6 and 43 days prior to returning the completed questionnaire. PROCEDURE: Veterinary practices were contacted weekly to obtain the names of clients whose pets had died until approximately 200 clients were identified. Clients were contacted by telephone, and a questionnaire designed to measure grief associated with pet death was mailed to those willing to participate within 1 to 14 days of their pet's death. The questionnaire measured potential correlates and modifiers of grief and included three outcome measures: social/emotional and physical consequences, thought processes, and despair. Demographic data were also collected. RESULTS: Approximately 30% of participants experienced severe grief. The most prominent risk factors for grief included level of attachment, euthanasia, societal attitudes toward pet death, and professional support from the veterinary team. CONCLUSIONS AND CLINICAL RELEVANCE: Bivariate and multivariate analyses highlighted the impact owners' attitudes about euthanasia and professional intervention by the veterinary team had on reactions to pet death. Owners' perceptions of societal attitudes, also a predictor of grief, indicate that grief for pets is different than grief associated with other losses.