Trends in companion animal access to veterinary care in Canada, 2007 to 2020.

Objective: Assess trends in access to veterinary care for companion animals in Canada. Procedure: Analysis and integration of available data, 2007 to 2020. Results: Cumulative growth in the Canadian veterinary workforce was 38%, and 49% for companion animal veterinarians. Clients per companion animal veterinarian decreased 30% from 2008 to 2020. Absolute client numbers increased 1.3%, compared to pet population growth of 17%. Medicalized pets (those that had received veterinary care in the past year) increased 25%, from 9.02 million in 2007 to 11.24 million in 2020. Non-medicalized pets increased 1.8%, from 4.48 million to 4.56 million. In 2007, 33% of pets were non-medicalized, compared to 29% (15% of dogs and 42% of cats) in 2020. There was a cumulative increase of 31% for total non-medicalized dogs, and a change of -5.6% for cats. Gross and net revenues per client increased by 99 and 112%, respectively, compared to cumulative inflation of 21%. Conclusion and clinical relevance: The analysis identified a large cohort of pets that had not received veterinary care each year. The trends were fewer clients per veterinarian, each paying higher veterinary costs, and suggested a relative, rather than absolute, veterinary capacity shortage overall. Accessible care-provision models must be encouraged, regulated for, and allowed to flourish alongside traditional models.

Tendances en matière d'accès des animaux de compagnie aux soins vétérinaires au Canada, 2007 à 2020. Objectif: Évaluer les tendances en matière d'accès aux soins vétérinaires pour les animaux de compagnie au Canada. Procédure: Analyse et intégration des données disponibles, 2007 à 2020. Résultats: La croissance cumulative de la main-d'œuvre vétérinaire canadienne était de 38 %, et de 49 % pour les médecins vétérinaires d'animaux de compagnie. Le nombre de clients par vétérinaire pour animaux de compagnie a diminué de 30 % entre 2008 et 2020. Le nombre absolu de clients a augmenté de 1,3 %, comparé à une croissance de la population d'animaux de compagnie de 17 %. Les animaux de compagnie médicalisés (ceux qui ont reçu des soins vétérinaires au cours de l'année écoulée) ont augmenté de 25 %, passant de 9,02 millions en 2007 à 11,24 millions en 2020. Les animaux de compagnie non médicalisés ont augmenté de 1,8 %, passant de 4,48 millions à 4,56 millions. En 2007, 33 % des animaux de compagnie étaient non médicalisés, contre 29 % (15 % des chiens et 42 % des chats) en 2020. Il y a eu une augmentation cumulée de 31 % pour le total des chiens non médicalisés, et une variation de ­5,6 % pour les chats. Les revenus bruts et nets par client ont augmenté respectivement de 99 et 112 %, par rapport à une inflation cumulée de 21 %. Conclusion et pertinence clinique: L'analyse a identifié chaque année une large cohorte d'animaux de compagnie qui n'avaient pas reçu de soins vétérinaires. Les tendances étaient moins de clients par vétérinaire, chacun payant des frais vétérinaires plus élevés, et suggéraient une pénurie globale de capacité vétérinaire relative plutôt qu'absolue. Les modèles de prestation de soins accessibles doivent être encouragés, réglementés et autorisés à prospérer aux côtés des modèles traditionnels.(Traduit par Dr Serge Messier).

A Pandora's box in feline medicine: presenting signs and surgical outcomes in 58 previously hoarded cats with chronic otitis media-interna.

OBJECTIVES: The aim of the present study was to report clinical findings, surgical complications and outcomes for previously hoarded cats treated surgically for otitis media-interna (OMI) and to investigate the risk factors for complications and poor outcomes. METHODS: A retrospective study was conducted of 58 cats from an institutional hoarding environment that underwent ventral bulla osteotomy (VBO). RESULTS: Inappetence was uncommon at presentation (9/58, 16%) compared with pruritus/alopecia (50%), nasopharyngeal signs (45%), otitis externa (OE) (79%) and otitis interna (OI) (ataxia ± head tilt/head excursions) in 40%. Purulent aural discharge occurred in 36% and polyps in 26%. The tympanic bulla wall was moderately or severely thickened radiographically in 38/108 (35%) ears. Cultures were positive for Streptococcus equi subspecies zooepidemicus in 26/48 (54%) cats. Of the 58 cats, 40 (69%) had complications after the first VBO and 19/30 (63%) after the second. Of 101 complications, 56 (55%), from 27/88 (31%) surgeries, were considered serious, including life-threatening perioperative complications in seven, OI in eight, prolonged anorexia in six and worsening of pruritus/alopecia in nine cases. Three cats developed xerostomia (dry mouth) after the second VBO. Pruritus/alopecia, nasopharyngeal signs, OE and purulent aural discharge resolved in a statistically significant proportion of cats but persisted in some. Full resolution of OI was uncommon. OI preoperatively, and surgery performed by a generalist (vs specialist) surgeon, were risk factors for OE at recheck (OI: odds ratio [OR] 4.35; 95% confidence interval [CI] 1.21-15.70; P = 0.02; surgery: OR 3.64; 95% CI 1.03-12.87; P = 0.045). No other prognostic indicators were identified. No variables tested were significantly associated with risk of serious complications or euthanasia. CONCLUSIONS AND RELEVANCE: Surgical management of chronic OMI was successful in most cases but was not benign and not always beneficial. The analysis was unable to identify clinically helpful outcome predictors. Optimal management of chronic feline OMI remains a challenge, particularly for animal shelters. Less invasive approaches and chronic medical management require further investigation.

Further variation of the adulticide protocol for the treatment of canine heartworm infection: can it be even shorter and cost less?

BACKGROUND: This retrospective study evaluated modified three-dose melarsomine treatment protocols in a shelter setting and compared them to the American Heartworm Society (AHS)-recommended protocol. METHODS: As compared with the AHS protocol, the shelter protocols utilized doxycycline 10 mg/kg once daily (SID) or twice daily (BID), and varied the time from initiation of doxycycline (day 1) to the first melarsomine injection (M1). Dogs were retrospectively grouped based on the shelter's current protocol (M1 on day 14; Group A) and the AHS protocol (M1 on day 60; Group C), allowing a week on either side of the target M1 day. Treatments that fell outside these ranges formed two additional treatment groups (Groups B and D). Respiratory complications were defined as respiratory signs requiring additional treatment, and were statistically compared for Groups A and C. New respiratory signs and gastrointestinal (GI) signs were compared between dogs receiving SID or BID doxycycline. RESULTS: One hundred fifty-seven dogs with asymptomatic or mild heartworm disease at presentation were included. All dogs survived to discharge. There was no statistically significant difference between Groups A (n = 79) and C (n = 27) for new respiratory signs post-melarsomine (P = 0.73). The time to M1 for 14 dogs that developed new respiratory signs was a median of 19 days, compared with 22 days for 143 dogs without new respiratory signs (P = 0.2). Respiratory complications post-melarsomine were uncommon. New respiratory signs post-melarsomine occurred in 10/109 (9.2%) dogs receiving SID doxycycline and 4/48 (8.3%) dogs receiving BID doxycycline (P > 0.999). GI signs prior to M1 were recorded for 40/109 (36.7%) dogs receiving SID doxycycline and 25/48 (52.1%) receiving BID doxycycline (P = 0.08). Forty-four follow-up antigen test results were available; all tests performed > 3 months after the third melarsomine injection were negative. CONCLUSIONS: This study provided support for initiating melarsomine after 14 days of doxycycline and for a lower doxycycline dose. Shorter and less expensive treatment protocols can increase lifesaving capacity and improve quality of life for shelter dogs by reducing the duration of exercise restriction and length of stay.

Efficacy of the Fast Track Friendship Group Program for Peer-rejected Children: A Randomized-Controlled Trial.

OBJECTIVE: To evaluate the benefits of the Fast Track Friendship Group program implemented as a stand-alone school-based intervention on the social cognitions, social behavior, peer and teacher relationships of peer-rejected students. METHOD: Over four successive years, 224 peer-rejected elementary students (57% White, 17% Black, 20% Latinx, 5% multiracial; 68% male; grades 1-4; Mage = 8.1 years old) were identified using peer sociometric nominations and randomly assigned to the intervention (n = 110) or a treatment-as-usual control group (n = 114). Four school districts serving economically-disadvantaged urban and rural communities participated. Intervention involved weekly small group social skills training with classmate partners, with sessions tailored to address individual child needs. Consultation meetings held at the start and mid-point of intervention were designed to help teachers and parents support the generalization of targeted skills. RESULTS: Multi-level linear models, with children nested within schools (controlling for demographics and baseline scores) documented improvements in social-cognitive skills (direct assessments of emotion recognition and competent social problem-solving), social behavior (teacher ratings of social skills and externalizing behavior), and interpersonal relationships (peer sociometric nominations of peer acceptance and friendships, teacher-rated student-teacher closeness). Significant effects were generally small (ds = .19-.36) but consistent across child sex, grade level, and behavioral characteristics. CONCLUSIONS: The intervention proved feasible for high-fidelity implementation in school settings and produced significant improvements in the social adjustment of peer-rejected children, validating the approach as a school-based Tier 2 intervention.

Behavior and adoptability of hoarded cats admitted to an animal shelter.

OBJECTIVES: The aim of this study was to analyze the behavioral characteristics and success of adoption for previously hoarded cats. METHODS: Shelter records and post-adoption surveys were analyzed for hoarded cats ⩾6 months old at intake. A non-standard scoring system was used. Intake scores were allocated contemporaneously and socialization scores were applied retrospectively for three time points (TPs): 5-10 days post-intake (shelter TP), ⩽1 week post-adoption (home TP1) and >1 week post-adoption (home TP2). Adoption returns were compared between hoarded and non-hoarded cats. RESULTS: The study included 195 hoarded cats, of which 174 were adopted. Of 164 cats with intake scores, 86 (52%) were scored as 'friendly' at intake. Forty-five cats had socialization scores for all of the TPs, and of these, the percentages of 'supersocial' or 'social' decreased from 87% at the shelter TP to 47% at home TP1, then increased to 84% at home TP2. Most cats that scored as 'tense' at intake had supersocial or social scores at home TP2. Nine of the 88 cats with survey results had out-of-box (OOB) elimination in either the shelter or home but only 1/88 in both. Adopters expressed positive feelings for 42/43 cats for which feelings-based language was used in their survey responses. Notable behaviors, such as neediness, were recorded for 48/88 cats. Relationships with other household pets were typically positive. Eighteen of 174 hoarded (10%) and 188/2662 non-hoarded (7.1%) cats were returned post-adoption. Of these, six hoarded and 87 non-hoarded returns included behavioral reasons. There were no significant differences between hoarded and non-hoarded cats for total or behavioral returns. CONCLUSIONS AND RELEVANCE: Hoarded cats had high adoption rates, high adopter satisfaction and the potential for good emotional well-being in adoptive homes. Behavior at intake and OOB elimination in the shelter may not reflect post-adoption behavior. Behavior-based outcome decisions for these vulnerable animals should be deferred to allow time for habituation.

Risk Factors for SARS-CoV-2 Infection and Illness in Cats and Dogs1.

We tested swab specimens from pets in households in Ontario, Canada, with human COVID-19 cases by quantitative PCR for SARS-CoV-2 and surveyed pet owners for risk factors associated with infection and seropositivity. We tested serum samples for spike protein IgG and IgM in household pets and also in animals from shelters and low-cost neuter clinics. Among household pets, 2% (1/49) of swab specimens from dogs and 7.7% (5/65) from cats were PCR positive, but 41% of dog serum samples and 52% of cat serum samples were positive for SARS-CoV-2 IgG or IgM. The likelihood of SARS-CoV-2 seropositivity in pet samples was higher for cats but not dogs that slept on owners' beds and for dogs and cats that contracted a new illness. Seropositivity in neuter-clinic samples was 16% (35/221); in shelter samples, 9.3% (7/75). Our findings indicate a high likelihood for pets in households of humans with COVID-19 to seroconvert and become ill.

Promoting Social and Emotional Learning in Middle School: Intervention Effects of Facing History and Ourselves.

There are fewer evidence-based social and emotional learning programs for middle school students compared to younger grades. This randomized controlled trial tests the effectiveness of Facing History and Ourselves (hereafter, Facing History) with a sample of 694 (Facing History n = 437; Comparison n = 257) students from a low-resourced school district. Youth self-identified as female (59%), Black/African American (61%), Hispanic/Latinx (18%), White (2%), and multi-racial or some other race/ethnicity (19%). Intervention students perceived their classrooms as more caring and democratic than students in the comparison group. They reported higher levels of empathy, prosocial behavior, and stronger participatory citizenship beliefs. This study expands the evidence-based of effective SEL programs available to schools.

Fecal viral DNA shedding following clinical panleukopenia virus infection in shelter kittens: a prospective, observational study.

OBJECTIVES: The aims of this study were to determine the magnitude and duration of fecal viral DNA shedding after diagnosis of feline panleukopenia (FP) in a group of shelter cats using quantitative real-time PCR (qPCR); to assess the utility of a negative point-of-care test or the resolution of diarrhea and systemic signs as proxy measures for qPCR positivity; and to investigate patterns of additional enteric pathogens in relation to feline panleukopenia viral shedding duration. METHODS: Feline panleukopenia virus (FPV) infection in clinically affected shelter cats was confirmed by a commercial qPCR test. Observations were made on days 0, 3, 7, 14 and 21 post-diagnosis. Fecal flotation, FPV qPCR and the canine parvovirus IDEXX SNAP Parvo ELISA (SNAP) test were performed on fecal samples. RESULTS: Forty cats and kittens with confirmed panleukopenia were initially enrolled. Sixteen kittens were sampled until day 14, and 12 were followed to day 21. Median DNA viral copy numbers fell below the diagnostic cut-off by day 7, with 13/16, 6/16, 1/16 and 0/12 testing PCR-positive on days 3, 7, 14 and 21, respectively. The SNAP test was positive in 12/16 kittens on day 0 and only 3/16 on day 3. SNAP test results, diarrhea and systemic signs were inconsistent in relation to qPCR positivity post-diagnosis. Additional enteric pathogens were common. The presence of additional pathogen types was suggestive of a longer PCR shedding duration, but this was not tested statistically owing to the small sample size. CONCLUSIONS AND RELEVANCE: These findings suggest that cats should be isolated for at least 14 days after a diagnosis of FP, but that release from isolation after this point is reasonable, in association with a multifaceted infection control strategy. The study findings did not support using SNAP test results, diarrhea or systemic signs as proxy measures for virus shedding.

Where Are We on the Animal Welfare Map? Using GIS to Assess Stakeholder Diversity and Inclusion.

Inequities exist in all facets of society, and animal welfare organizations (AWOs) and their communities are no exception. These organizations interface with multiple stakeholder groups. An active analysis of stakeholder groups to identify under-served areas and communities has not been performed. Using stakeholder data from Toronto Humane Society (THS) from 2015-2019, this study performed a retrospective spatial analysis to identify well served and under-served geographic areas for adopters, surrenders, public veterinary service (PVS) clients, volunteers and foster parents, using Hot Spot analysis. Correlation analysis was performed to determine whether the spatial distribution of the groups correlated with the four socioeconomic metrics of the 2016 Ontario Marginalization Index (residential instability, material deprivation, dependency, and ethnic concentration), and a metric representing the distribution of Indigenous residents. For each stakeholder group, there were well served areas, typically in central Toronto where THS is located, and under-served areas, typically in the north-west and north-east corners of Toronto and in the surrounding cities of the Greater Toronto Area. The area served by THS PVS extended further north than the other hot spot areas. The number of adopters increased as the residential instability metric increased, whereas the number of adopters decreased as the ethnic concentration metric increased. The rate of surrenders increased as the Indigenous metric increased. Public Veterinary Service clients increased as the residential instability, material deprivation, and Indigenous metrics increased. One of the primary limitations of this study was the confounding factor of distance from THS. Individuals living further from THS are less likely to utilize its services, particularly if there is another accessible AWO nearby, and therefore may appear to reflect an under-served population that may not truly be under-served. A regional approach would help to overcome this limitation. The results provide useful insights into stakeholder engagement and provide a foundation for analysis of more targeted areas, as well as for strategies to reach under-served demographics. Similar analyses by other AWOs would be helpful to address inequities in a larger geographic area. Animal welfare organizations can improve program effectiveness by adding data analytics skills to the more traditional skills associated with this sector.

An Accessible Alternative to Melarsomine: "Moxi-Doxy" for Treatment of Adult Heartworm Infection in Dogs.

Canine heartworm infection, caused by the filarial parasite Dirofilaria immitis, represents a serious and expanding animal welfare concern that is expected to increase due to the effects of climate change and the COVID-19 pandemic. A body of evidence has emerged to support the use of a non-arsenical adulticide treatment protocol, using moxidectin and doxycycline to kill adult heartworms over a prolonged period. While a three-dose protocol using the arsenical drug melarsomine is currently the safest and most effective treatment for heartworm infection, this drug is not available in some countries and is inaccessible for many owners and animal shelters. Moxidectin-doxycycline (moxi-doxy) provides a viable alternative to no treatment at all, in cases where arsenical treatment is not possible. Based on current evidence, the most effective non-arsenical treatment regimen is doxycycline 10 mg/kg PO q 12 or 24 h for 28 days, combined with topical moxidectin at label dose. Moxidectin is repeated monthly until no antigen detected (NAD) status is confirmed. Sustained release injectable moxidectin, in combination with doxycycline, may provide an alternative in remote regions or in settings where significant compliance or accessibility concerns exist, but more studies are needed. In moxi-doxy protocols, doxycycline should be repeated annually until NAD. This review summarizes the safety and efficacy of moxi-doxy, addresses controversies surrounding this treatment approach, and provides detailed recommendations for treatment regimens and post-treatment testing.

Diagnostic testing for feline panleukopenia in a shelter setting: a prospective, observational study.

OBJECTIVES: The aim of this study was to optimize the diagnosis of feline panleukopenia virus (FPV) infection in a shelter setting by: (1) comparing the results of the canine parvovirus IDEXX SNAP Parvo (SNAP) point-of-care ELISA with a commercial FPV quantitative real-time PCR (qPCR) test; (2) assessing whether vomit and anal/rectal swabs could be used for early diagnosis; and (3) clarifying the interpretation of weak-positive SNAP test results. METHODS: The study included shelter cats and kittens with incomplete or unknown vaccination history that had clinical signs suspicious for feline panleukopenia and fecal SNAP and PCR tests performed within 24 h of onset. Feces, anal/rectal swabs and vomit were tested using SNAP and PCR, with fecal PCR utilized as the reference standard. RESULTS: One hundred and forty-five cats were included. Seventeen were diagnosed with FPV infection and 62 were negative; 66 could not be individually designated because they were co-housed. Sensitivity was as follows: fecal SNAP 55% (n = 102; 95% confidence interval [CI] 32-77); swab SNAP 30% (n = 55; 95% CI 7-65); swab PCR 77% (n = 55; 95% CI 46-95); and vomit PCR 100% (n = 17; 95% CI 16-100). Specificity was high (96-100%) for all sample and test types. For PCR-positive fecal samples, true-positive SNAP tests (including weak positives) had significantly higher DNA viral copy numbers than false-negative SNAP tests (P = 0.0031). CONCLUSIONS AND RELEVANCE: The SNAP ELISA should be viewed as an initial diagnostic test to rule in feline panleukopenia. Positive fecal SNAP test results, including weak positives, are highly likely to be true positives in clinically affected animals. Negative results in clinically affected animals are unreliable and should be followed up with PCR testing.

Antithrombin deficiency: A pediatric disorder.

INTRODUCTION: Hereditary antithrombin (AT) deficiency is an autosomal dominant thrombophilic disorder. Guidelines do not support routine testing of children based on personal or familial thrombosis. AIM: To investigate clinical, genetic and laboratory profiles of AT deficient children and their affected family members. MATERIALS AND METHODS: Data were analyzed from a prospective cohort of pediatric patients with AT deficiency. The SERPINC1 gene was sequenced for all individuals with available DNA. AT, thromboelastography (TEG), calibrated automated thrombogram (CAT), D-dimer, thrombin-antithrombin complex (TAT) and factor VIII activity were performed on patient samples. RESULTS: Thirty-six individuals from 11 families had AT deficiency (activities 45-70 U/dL) with incident thrombosis in 13 children and 10 adults (64% overall). Three neonates presented with middle cerebral artery and/or aortic occlusions with inferior vena cava and cerebral or renal vein thromboses in 2 of the 3. Two pre-pubertal children were symptomatic, one with cerebral venous sinus thrombosis who suffered recurrent arterial and venous thrombi. Both Type I and Type II AT deficiencies conferred a high severity of thromboses. Heterozygous SERPINC1 mutations were identified in seven families; three were novel, resulting in missense, splice site and frameshift alterations. Thrombin generation (CAT) was increased in all asymptomatic affected patients including 9 children and 1 adult. CONCLUSIONS: Genetic AT deficiency often presents in infants and children, warranting laboratory evaluation based on personal and family history. Increased thrombin generation was detected in all asymptomatic children and adults, suggesting a possible role in detecting and monitoring individuals at risk for thrombosis.

Teacher, Center, and Neighborhood Characteristics Associated with Variations in Preschool Quality in Childcare Centers.

BACKGROUND: Childcare programs serving preschool children are generally of poorer quality than publicly-funded preschools both in terms of their classroom processes and structural features. Research on childcare programs has typically collapsed them into a single group, yet these programs vary greatly in neighborhood disadvantage and organization as they are managed by for-profit chains, non-profit community organizations, faith-based organizations, or individual owners. Little is known about variations in childcare program quality and what factors are associated with quality. OBJECTIVE: The current study utilized latent profile analysis (LPA) with classroom process, structural features, and neighborhood disadvantage indicators to identify patterns of quality and neighborhood disadvantage within a diverse sample of childcare programs serving preschool children. METHODS: Classroom processes (instructional support, emotional support, classroom management, positive discipline) and structural features (teacher age, experience, education, and satisfaction) data was collected from preschool teachers (N = 127) from 76 childcare programs. Neighborhood disadvantage (median income; rates of unemployment, single parents, and education) was measured using census tract data. RESULTS: LPA indicated two profiles of childcare programs with high-quality classroom processes and two with poorer processes. Both of the high-quality profiles were in low-to-moderate income neighborhoods and the two low-quality profiles were in more affluent neighborhoods. Subsequent analyses suggested quality covaried with management type with the lowest quality centers often run by for-profit chains. CONCLUSIONS: Connections between classroom processes, structural features, and neighborhood disadvantage are complex making it extremely challenging for parents to identify high-quality care for their children.

Prevalence of heartworm in relocated, local and outreach clinic dogs: A Canadian sheltering perspective.

This study reports heartworm (Dirofilaria immitis) prevalence in dogs tested by an animal shelter located in a low-prevalence region of Ontario, Canada. From 2015-2018, 4567 unique dogs were tested. The prevalence of heartworm was 3.9 %, with sub-prevalence of 0.3 % (2/662) for dogs surrendered within the Greater Toronto Area (both dogs were originally imported from the US); 6.6 % (130/1,981) for dogs relocated from beyond the Greater Toronto Area; 0% (0/1,668) for dogs tested at the shelter's public veterinary services clinic and 18.4 % (47/256) for owned dogs tested at outreach clinics at a First Nations community in south-eastern Ontario. More than half (54.7 %) of the heartworm-positive dogs originated from Canada. Most heartworm-positive dogs from the US (72/80; 90 %) were transferred from Texas and Georgia. Ninety-three heartworm-positive Canadian dogs were from Ontario, 4 from Manitoba and 1 from Quebec. Most (83/98, 84.7 %) were from four First Nations communities. The prevalence in homeless dogs from one First Nations community in south-western Ontario was 36.5 % (31/85). For 140 shelter dogs with at least one positive test result, there was 91 % concordance between shelter and reference laboratory antigen tests and poor agreement between antigen tests and microfilarial tests (approximately 50 %). Results of historical tests and post-relocation antigen tests were discordant in 28.2 % (shelter point-of-care tests) and 36.2 % (reference laboratory tests) of cases. This was most commonly due to negative historical results followed by positive results after relocation. Microfilarial filtration tests were positive for 77.1 % antigen-positive dogs from First Nations communities, compared with 37.1 % of dogs from other sources. Microfilarial counts were significantly higher for dogs from First Nations communities. This study demonstrated endemic and hyperendemic foci of heartworm within Canada, which were presumed to be associated with limited access to veterinary care. The results support recommendations to retest previously negative animals after relocation.

Cu2+-based distance measurements by pulsed EPR provide distance constraints for DNA backbone conformations in solution.

Electron paramagnetic resonance (EPR) has become an important tool to probe conformational changes in nucleic acids. An array of EPR labels for nucleic acids are available, but they often come at the cost of long tethers, are dependent on the presence of a particular nucleotide or can be placed only at the termini. Site directed incorporation of Cu2+-chelated to a ligand, 2,2'dipicolylamine (DPA) is potentially an attractive strategy for site-specific, nucleotide independent Cu2+-labelling in DNA. To fully understand the potential of this label, we undertook a systematic and detailed analysis of the Cu2+-DPA motif using EPR and molecular dynamics (MD) simulations. We used continuous wave EPR experiments to characterize Cu2+ binding to DPA as well as optimize Cu2+ loading conditions. We performed double electron-electron resonance (DEER) experiments at two frequencies to elucidate orientational selectivity effects. Furthermore, comparison of DEER and MD simulated distance distributions reveal a remarkable agreement in the most probable distances. The results illustrate the efficacy of the Cu2+-DPA in reporting on DNA backbone conformations for sufficiently long base pair separations. This labelling strategy can serve as an important tool for probing conformational changes in DNA upon interaction with other macromolecules.

Medical conditions and outcomes in 371 hoarded cats from 14 sources: a retrospective study (2011-2014).

OBJECTIVES: The aims of this study were to: (1) describe the source, route of surrender and signalment of hoarded cats relinquished to the Toronto Humane Society (THS); (2) document the prevalence of medical conditions by group (place of origin); (3) compare medical conditions between institutional hoarding (IH) and non-institutional hoarding (NIH) environments; and (4) report length of stay (LOS) and outcomes in hoarded and non-hoarded cats. METHODS: A retrospective, descriptive epidemiological study was performed using THS records from between July 2011 and June 2014. The prevalence of medical conditions was calculated for the different groups. Univariable logistic regression with a random intercept to account for autocorrelation among animals from the same group was used to examine the influence of IH and NIH environments on selected medical conditions. LOS and outcomes were calculated for hoarded and non-hoarded cats. RESULTS: Three hundred and seventy-one hoarded cats from 14 sources were included. The majority (n = 352/371) were surrendered voluntarily, many with the assistance of a community intermediary. Upper respiratory infection (URI) was the most common medical condition (38% of cats), followed by dermatological disease (30%). The prevalence of medical conditions varied substantially between groups. The odds of URI at intake (odds ratio [OR] 4.35, P = 0.044) and chronic URI (OR 23.70, P <0.0001) were significantly greater for IH compared with NIH. Adoption rates, euthanasia rates and LOS were similar for hoarded and non-hoarded cats. CONCLUSIONS AND RELEVANCE: The different prevalence of medical conditions in groups of hoarded cats indicates a continuum of harm and severity in animal hoarding. Hoarded cats can have LOS and live release rates comparable with non-hoarded cats. Cats from IH were significantly more likely to have chronic URI. This study highlights the need for a greater focus on IH, as well as the role of community intermediaries and the potential for a harm reduction approach to animal hoarding.

Factor VIII prophylaxis effects outweigh other hemostasis contributors in predicting severe haemophilia A joint outcomes.

INTRODUCTION: The Joint Outcome Study (JOS) demonstrated that previously untreated children with severe haemophilia A treated with prophylactic factor VIII (FVIII) concentrate had superior joint outcomes at age 6 years compared to those children treated episodically for bleeding. However, variation in joint outcome within each treatment arm was not well explained. AIM: In this study, we sought to better understand variation in joint outcomes at age 6 years in participants of the JOS. METHODS: We evaluated the influence of FVIII half-life, treatment adherence, constitutional coagulant and anticoagulant proteins, and global assays on joint outcomes (number of joint bleeds, total number of bleeds, total MRI score and joint physical exam score). Logistic regression was used to evaluate the association of variables with joint failure status on MRI, defined as presence of subchondral cyst, surface erosion or joint-space narrowing. Each parameter was also correlated with each joint outcome using Spearman correlations. RESULTS: Prophylaxis treatment arm and FVIII trough were each found to reduce risk of joint failure on univariate logistic regression analysis. When controlling for treatment arm, FVIII trough was no longer significant, likely because of the high level of covariation between these variables. We found no consistent correlation between any laboratory assay performed and any joint outcome parameter measured. CONCLUSION: In the JOS, the effect of prescribed prophylactic FVIII infusions on joint outcome overshadowed the contribution of treatment adherence, FVIII half-life, global assays of coagulation and constitutional coagulation proteins. (ClinicalTrials.gov number, NCT00207597).

Identification and characterization of novel mutations implicated in congenital fibrinogen disorders.

INTRODUCTION: Fibrinogen is a complex molecule comprised of two sets of Aα, Bß, and γ chains. Fibrinogen deficiencies can lead to the development of bleeding or thromboembolic events. The objective of this study was to perform DNA sequence analysis of patients with clinical fibrinogen abnormalities, and to perform genotype-phenotype correlations. MATERIALS AND METHODS: DNA from 31 patients was sequenced to evaluate disease-causing mutations in the three fibrinogen genes: FGA,FGB, and FGG. Clinical data were extracted from medical records or from consultation with referring hematologists. Fibrinogen antigen and functional (Clauss method) assays, as well as reptilase time (RT) and thrombin time (TT) were obtained for each patient. Molecular modeling was used to simulate the functional impact of specific missense variants on the overall protein structure. RESULTS: Seventeen mutations, including six novel mutations, were identified in the three fibrinogen genes. There was little correlation between genotype and phenotype. Molecular modeling predicted a substantial conformational change for a novel variant, FGG p.Ala289Asp, leading to a more rigid molecule in a region critical for polymerization and alignment of the fibrin monomers. This mutation is associated with both bleeding and clotting in the two affected individuals. CONCLUSIONS: Robust genotype-phenotype correlations are difficult to establish for fibrinogen disorders. Molecular modeling might represent a valuable tool for understanding the function of certain missense fibrinogen mutations but those should be followed by functional studies. It is likely that genetic and environmental modifiers account for the incomplete penetrance and variable expressivity that characterize fibrinogen disorders.

Assessment of real-time PCR cycle threshold values in Microsporum canis culture-positive and culture-negative cats in an animal shelter: a field study.

Objectives Real-time PCR provides quantitative information, recorded as the cycle threshold (Ct) value, about the number of organisms detected in a diagnostic sample. The Ct value correlates with the number of copies of the target organism in an inversely proportional and exponential relationship. The aim of the study was to determine whether Ct values could be used to distinguish between culture-positive and culture-negative samples. Methods This was a retrospective analysis of Ct values from dermatophyte PCR results in cats with suspicious skin lesions or suspected exposure to dermatophytosis. Results One hundred and thirty-two samples were included. Using culture as the gold standard, 28 were true positives, 12 were false positives and 92 were true negatives. The area under the curve for the pretreatment time point was 96.8% (95% confidence interval [CI] 94.2-99.5) compared with 74.3% (95% CI 52.6-96.0) for pooled data during treatment. Before treatment, a Ct cut-off of <35.7 (approximate DNA count 300) provided a sensitivity of 92.3% and specificity of 95.2%. There was no reliable cut-off Ct value between culture-positive and culture-negative samples during treatment. Ct values prior to treatment differed significantly between the true-positive and false-positive groups ( P = 0.0056). There was a significant difference between the pretreatment and first and second negative culture time points ( P = 0.0002 and P <0.0001, respectively). However, there was substantial overlap between Ct values for true positives and true negatives, and for pre- and intra-treatment time points. Conclusions and relevance Ct values had limited usefulness for distinguishing between culture-positive and culture-negative cases when field study samples were analyzed. In addition, Ct values were less reliable than fungal culture for determining mycological cure.