Feature weighted models to address lineage dependency in drug-resistance prediction from Mycobacterium tuberculosis genome sequences.

MOTIVATION: Tuberculosis (TB) is caused by members of the Mycobacterium tuberculosis complex (MTBC), which has a strain- or lineage-based clonal population structure. The evolution of drug-resistance in the MTBC poses a threat to successful treatment and eradication of TB. Machine learning approaches are being increasingly adopted to predict drug-resistance and characterize underlying mutations from whole genome sequences. However, such approaches may not generalize well in clinical practice due to confounding from the population structure of the MTBC. RESULTS: To investigate how population structure affects machine learning prediction, we compared three different approaches to reduce lineage dependency in random forest (RF) models, including stratification, feature selection, and feature weighted models. All RF models achieved moderate-high performance (area under the ROC curve range: 0.60-0.98). First-line drugs had higher performance than second-line drugs, but it varied depending on the lineages in the training dataset. Lineage-specific models generally had higher sensitivity than global models which may be underpinned by strain-specific drug-resistance mutations or sampling effects. The application of feature weights and feature selection approaches reduced lineage dependency in the model and had comparable performance to unweighted RF models. AVAILABILITY AND IMPLEMENTATION: https://github.com/NinaMercedes/RF_lineages.

Bayesian phase II adaptive randomization by jointly modeling efficacy and toxicity as time-to-event outcomes.

The main goals of Phase II trials are to identify the therapeutic efficacy of new treatments and continue monitoring all the possible adverse effects. In Phase II trials, it is important to develop an adaptive randomization (AR) procedure that takes into account both the efficacy and toxicity. In most existing articles, toxicity is modeled as a binary endpoint through an unobservable random effect (frailty) to link the efficacy and toxicity. However, this approach does not capture toxicity profiles that evolve over time. In this article, we propose a new Bayesian adaptive randomization (BAR) procedure using the covariate-adjusted efficacy-toxicity ratio (ETR) index, where efficacy and toxicity are jointly modelled as time-to-event (TTE) outcomes. Furthermore, we also propose early stopping rules for toxicity and futility such that inferior treatments can be dropped at earlier time of trial. Simulation results show that compared to the BAR procedures based solely on the efficacy and that based on TTE efficacy and binary toxicity outcomes, the proposed BAR procedure can better identify the difference in treatment toxicity such that it can assign more patients to the superior treatment arm under some scenarios.

Rh proteins and H+ transporters involved in ammonia excretion in Amur Ide (Leuciscus waleckii) under high alkali exposure.

High alkaline environment can lead to respiratory alkalosis and ammonia toxification to freshwater fish. However, the Amur ide (Leuciscus waleckii), which inhabits an extremely alkaline lake in China with titratable alkalinity up to 53.57 mM (pH 9.6) has developed special physiological and molecular mechanisms to adapt to such an environment. Nevertheless, how the Amur ide can maintain acid-base balance and perform ammonia detoxification effectively remains unclear. Therefore, this study was designed to study the ammonia excretion rate (Tamm), total nitrogen accumulation in blood and tissues, including identification, expression, and localization of ammonia-related transporters in gills of both the alkali and freshwater forms of the Amur ide. The results showed that the freshwater form Amur ide does not have a perfect ammonia excretion mechanism exposed to high-alkaline condition. Nevertheless, the alkali form of Amur ide was able to excrete ammonia better than freshwater from Amur ide, which was facilitated by the ionocytes transporters (Rhbg, Rhcg1, Na+/H+ exchanger 2 (NHE2), and V-type H+ ATPase (VHA)) in the gills. Converting ammonia into urea served as an ammonia detoxication strategy to reduced endogenous ammonia accumulation under high-alkaline environment.

A direct healthcare cost analysis of recombinant LH versus hMG supplementation on FSH during controlled ovarian hyperstimulation in the GnRH-antagonist protocol.

PURPOSE: We have previously published a retrospective matched-case control study comparing the effect of recombinant LH (r-hLH) versus highly purified human menopausal gonadotropin (hMG) supplementation on the follicle-stimulating hormone (FSH) during controlled ovarian hyperstimulation (COH) in the GnRH-antagonist protocol. The result from that study showed that the cumulative live birth rate (CLBR) was significantly higher in the r-hLH group (53% vs. 64%, p = 0.02). In this study, we aim to do a cost analysis between these two groups based on our previous study. METHODS: The analysis consisted of 425 IVF and ICSI cycles in our previous study. There were 259 cycles in the r-hFSH + hMG group and 166 cycles in the r-hFSH + r-hLH group. The total cost related to the treatment of each patient was recorded. Probabilistic sensitivity analysis (PSA) and a cost-effectiveness acceptability curve (CEAC) were performed and created. RESULTS: The total treatment cost per patient was significantly higher in the r-hFSH + r-hLH group than in the r-hFSH + hMG group ($4550 ± 798.86 vs. $4290 ± 734.6, p = 0.003). However, the mean cost per live birth in the r-hFSH + hMG group was higher at $8052, vs. $7059 in the r-hFSH + r-hLH group. The CEAC showed that treatment with hFSH + r-hLH proved to be more cost-effective than treatment with r-hFSH + hMG. Willingness-to-pay was evident when considering a hypothetical threshold of $18,513, with the r-hFSH + r-hLH group exhibiting a 99% probability of being considered cost-effective. CONCLUSION: The cost analysis showed that recombinant LH is more cost-effective than hMG supplementation on r-hFSH during COH in the GnRH-antagonist protocol.

Systematic analysis reveals that colony housing aligns gait profiles and strengthens link between histological and micro-CT bone markers in rat models of osteoarthritis.

Osteoarthritis (OA) etiopathogenesis is complex with strong environmental/lifestyle determinants that, in laboratory animals, extend to social context and stress levels. This study seeks to identify whether colony housing of rats exerts a social impact on locomotion behaviors to influence alignment between symptomatic (gait) and structural (bone micro-CT measures, cartilage morphometry, and histology) OA outcome measures. Rats were randomly allocated to conventional (type IV; n = 48) or rat colony cage (RCC; n = 30) housing, further randomized to OA surgical models (ACLT + tMx, MMT or DMM) or no surgery (control), and maintained for 19 weeks during which multiple gait recordings were made. Standard histological grading and bone micro-CT data were collected at necropsy. Principal component analysis was used to summarize the variation in gait, micro-CT or histology. Linear mixed effects model or two-way ANOVA was employed to evaluate the impact of the housing system, surgery and time on gait, or micro-CT and histology components Analyses reveal that RCC exaggerates trends in gait change via a combined effect of the housing system and surgery. Intriguingly, RCC-housed nonoperated control rats showed similar gait changes to rats subjected to surgery; the latter exhibited significant structural joint changes in both systems. Stronger correlation between histological and micro-CT bone changes were found in medial and lateral tibia joint compartments of rats housed in RCC system. This study has established that rat social housing exaggerates outcomes in traditional histological measures of OA, generates stronger links between histology and micro-CT bone changes and removes gait differences as a variable in their etiology.

Fox (Vulpes vulpes) involvement identified in a series of cat carcass mutilations.

This study was designed to identify the cause of mutilation and death in 32 cats, part of a larger cohort found dead in Greater London, the United Kingdom, between 2016 and 2018. At the time, discussion in the media led to concerns of a human serial cat killer (dubbed The Croydon Cat Killer) pursuing domestic cats, causing a state of disquietude. Given the link between animal abuse and domestic violence, human intervention had to be ruled out. Using a combination of DNA testing, computed tomography imaging, and postmortem examination, no evidence was found to support any human involvement. Instead, a significant association between cat carcass mutilation and the presence of fox DNA was demonstrated. Gross examination identified shared characteristics including the pattern of mutilation, level of limb or vertebral disarticulation, wet fur, wound edges with shortened fur, and smooth or irregular contours, and marks in the skin, muscle, and bone consistent with damage from carnivore teeth. Together these findings supported the theory that the cause of mutilation was postmortem scavenging by red foxes (Vulpes vulpes). The probable cause of death was established in 26/32 (81%) carcasses: 10 were predated, 8 died from cardiorespiratory failure, 6 from blunt force trauma, one from ethylene glycol toxicity, and another from liver failure. In 6 carcasses a cause of death was not established due to autolysis and/or extensive mutilation. In summary, this study highlights the value of a multidisciplinary approach to fully investigate cases of suspected human-inflicted mutilation of animals.

Weighting the relative importance of behaviors affecting gait score.

This research communication evaluates experts' opinions on the importance and weights of six gait aspects. In 2016, a Qualtrics (Qualtrics LLC., Provo, Utah) survey was distributed to lameness experts. Six gait aspects - general symmetry, tracking, spine curvature, head bobbing, speed and abduction as well as adduction were included. Respondents were asked to rank the gait aspects from 1 (most important) to 6 (least important), and to indicate which weight each gait aspect should receive when assessing lameness. For each gait aspect, frequency (percentage %) was used to describe the distribution of rank, and medians as well as 25th and 75th percentiles were used to summarize assigned weights. Thirty-nine percent of respondents ranked general symmetry first, followed by 32% for tracking, and 19% ranked spine curvature third. Head bobbing ranked fourth with 10% whereas, speed, abduction and adduction were not ranked. The median, 25th and 75th percentiles weight for each gait aspect were: general symmetry (25, 15, and 30), tracking (20, 10, and 30), spine curvature (20, 10, and 21), head bobbing (15, 10, and 20), speed (10, 5, and 20), and abduction and adduction (10, 5, and 10). General symmetry and tracking were deemed the most important gait aspects. A composite gait score can be calculated based on weighted importance of different gait aspects to indicate possible lameness.

The effect of weather, air pollution and seasonality on the number of patient visits for epileptic seizures: A population-based time-series study.

OBJECTIVE: The objective of the study was to explore the influences of seasonality, meteorological conditions, and air pollution exposure on the number of patients who visit the hospital due to seizures. METHODS: Outpatient and inpatient data from the National Health Insurance Database of Taiwan from 2009 to 2013, meteorological data from the Meteorological Bureau, and air pollution exposure data from the Taiwan Air Quality Monitoring Stations were collected and integrated into daily time series data. The following data processing and analysis results are based on the mean of the 7 days' lag data of the 18 meteorological condition/air pollution exploratory factors to identify the critical meteorological conditions and air pollution exposure factors by executing univariate analysis. The average hospital visits for seizure per day by month were used as an index of observation. The effect of seasonality has also been examined. RESULTS: The average visits per day by month had a significant association with 10 variables. Overall, the number of visits due to these factors has been estimated to be 71.529 (13.7%). The most obvious factors affecting the estimated number of visits include ambient temperature, CH4, and NO. Six air pollutants, namely CH4, NO, CO, NO2, PM2.5, and NMHC had a significantly positive correlation with hospital visits due to seizures. Moreover, the average daily number of hospital visits was significantly high in January and February (winter season in Taiwan) than in other months (R2 = 0.422). CONCLUSION: The prediction model obtained in this study indicates the necessity of rigorous monitoring and early warning of these air pollutants and climate changes by governments. Additionally, the study provided a firm basis for establishing prediction models to be used by other countries or for other diseases.

A RETROSPECTIVE STUDY OF BITE WOUND MANAGEMENT IN RING-TAILED LEMURS (LEMUR CATTA) HOUSED WITHIN FIVE BRITISH ZOOS.

Captive nonhuman primates frequently experience bite wounds inflicted by conspecifics; however, few studies evaluate wound management in these animals. This study evaluates bite wound management in 35 captive ring-tailed lemurs (RTL, Lemur catta) held within five British zoological institutions between 2015 and 2019. Treatments for 152 bite incidents resulting in 211 bite wounds were summarized. Treatment efficacy and potential risk factors for bite wound complications were evaluated for correlations with outcomes. Treatments in the first 48 hours included analgesia alone (54/152; 35.5%); analgesia and antimicrobials (42/152; 27.6%); no treatment (26/152; 17.1%); analgesia, antimicrobials, and wound management under anesthesia (24/152; 15.8%); and antimicrobials alone (6/152; 3.9%). Poor outcomes were observed in RTL in association with 20.4% of wounds (43/211): signs of pain (SOP) after 48 hours were the most common (30/211; 14.2%), followed by signs of infection (SOI, 16/211; 7.6%), wound dehiscence (7/211; 3.3%), and abnormal function or behavior at day 30 (4/ 211; 1.9%). Poor outcomes were more likely with severe bite wounds and bite wounds to the hand or limb (thoracic or pelvic, excluding the hand or foot). Specifically, SOP were more likely to be observed with severe wounds and solitary wounds, and SOI were more likely to be observed with wounds not treated with early antimicrobials. When SOI occurred, most resolved with antimicrobial treatment alone. Early analgesia is recommended for RTL with bite wounds. If SOP persist, multimodal analgesia should be administered. Clinicians should consider withholding antimicrobials for RTL with bite wounds of mild and moderate severity until SOI are observed.

The Global Health Security index and Joint External Evaluation score for health preparedness are not correlated with countries' COVID-19 detection response time and mortality outcome.

Global Health Security Index (GHSI) and Joint External Evaluation (JEE) are two well-known health security and related capability indices. We hypothesised that countries with higher GHSI or JEE scores would have detected their first COVID-19 case earlier, and would experience lower mortality outcome compared to countries with lower scores. We evaluated the effectiveness of GHSI and JEE in predicting countries' COVID-19 detection response times and mortality outcome (deaths/million). We used two different outcomes for the evaluation: (i) detection response time, the duration of time to the first confirmed case detection (from 31st December 2019 to 20th February 2020 when every country's first case was linked to travel from China) and (ii) mortality outcome (deaths/million) until 11th March and 1st July 2020, respectively. We interpreted the detection response time alongside previously published relative risk of the importation of COVID-19 cases from China. We performed multiple linear regression and negative binomial regression analysis to evaluate how these indices predicted the actual outcome. The two indices, GHSI and JEE were strongly correlated (r = 0.82), indicating a good agreement between them. However, both GHSI (r = 0.31) and JEE (r = 0.37) had a poor correlation with countries' COVID-19-related mortality outcome. Higher risk of importation of COVID-19 from China for a given country was negatively correlated with the time taken to detect the first case in that country (adjusted R2 = 0.63-0.66), while the GHSI and JEE had minimal predictive value. In the negative binomial regression model, countries' mortality outcome was strongly predicted by the percentage of the population aged 65 and above (incidence rate ratio (IRR): 1.10 (95% confidence interval (CI): 1.01-1.21) while overall GHSI score (IRR: 1.01 (95% CI: 0.98-1.01)) and JEE (IRR: 0.99 (95% CI: 0.96-1.02)) were not significant predictors. GHSI and JEE had lower predictive value for detection response time and mortality outcome due to COVID-19. We suggest introduction of a population healthiness parameter, to address demographic and comorbidity vulnerabilities, and reappraisal of the ranking system and methods used to obtain the index based on experience gained from this pandemic.

The effect of visitors on the behavior of zoo-housed western lowland gorillas (Gorilla gorilla gorilla).

Primates, especially apes, are popular with the public, often attracting large crowds. These crowds could cause behavioral change in captive primates, whether positive, neutral, or negative. We examined the impact of visitors on the behavior of six western lowland gorillas (Gorilla gorilla gorilla), observing the troop over 6 weeks during high season (4.5 hr/day, 35 days, May-July 2016). We used focal scan sampling to determine activity budget and enclosure usage, and focal continuous sampling to identify bouts of anxiety-related behavior (visitor-directed vigilance, self-scratching, and aggression). Both daily zoo-entry numbers (VGATE ) and instantaneous crowds at the exhibit (VDENSITY ) were measured. Overall, VGATE had little effect across behaviors. However, consistent with the more acute time frame of measurement, VDENSITY was a better predictor of behavior; at high crowd volumes, we observed significant group-level changes in activity budget (increased inactivity, increased locomotion, and decreased environment-related behaviors), increase in some anxiety-related behaviors, and decreased enclosure usage. Although contributing similar effects, it could not be determined if crowd numbers, composition, or noise most affected the troop, nor any chronic effects of exposure to large crowds. Nevertheless, our findings suggest that measures to minimize the impacts of large crowds at the exhibit would be beneficial. Furthermore, we highlight potential discrepancies between common methods for measuring visitor numbers: VGATE is less sensitive to detecting visitor effects on behavioral indices than VDENSITY . Future studies should appropriately match the biological time frame of welfare indicators and visitor measures used to ensure the reliability of findings.

Nonocular Melanocytic Neoplasia in Cats: Characterization and Proposal of a Histologic Classification Scheme to More Accurately Predict Clinical Outcome.

Nonocular melanocytic neoplasia is considered uncommon in cats yet is routinely encountered in diagnostic pathology and recognized to exhibit a wide variation in biological behavior. Accurate prediction of clinical outcomes is challenging with no widely recognized prognostic criteria. Signalment and tumor location were retrospectively evaluated in 324 cats diagnosed with nonocular melanocytic neoplasia. Histologic features were described in 141 neoplasms and outcome data were available in 79 cases. Immunohistochemistry using Melan-A, PNL-2, cyclooxygenase 2 (COX-2), and E-cadherin was performed in a subset (n = 24). Multivariate analysis identified tumor site, mitotic count, and the presence of intratumoral necrosis to be independent predictors of tumor-related death. On the basis of these findings, we propose a novel histologic grading scheme in which nonocular melanocytic neoplasms involving the lips, oral or nasal mucosa, or nasal planum are considered high grade if they fulfill 1 or both of the following criteria: at least 4 mitoses in 10 high-power fields (HPF) or presence of intratumoral necrosis; those arising elsewhere are considered high grade if they fulfill both of the above criteria. Of 79 tumors with outcome data, 43 (54%) were low grade and 36 (46%) were high grade. The grading system had an 80% sensitivity and 92% specificity for predicting tumor-related death in this population of cats. Median survival for cats with low-grade tumors was not reached, and the median survival was 90 days for those with a high-grade tumor. PNL-2 and Melan-A were sensitive markers for feline nonocular melanocytic neoplasia, and although not significantly associated with prognosis, a large proportion expressed COX-2, suggesting a potential therapeutic role for COX-2 inhibitors.

Visual outcome following phacoemulsification in English Cocker Spaniels with suspected progressive retinal atrophy: A retrospective multicenter study of 54 cases (2002-2017).

OBJECTIVE: To describe the visual outcome following phacoemulsification in English Cocker Spaniels (ECS) affected by cataracts and suspected progressive retinal atrophy (PRA). ANIMALS STUDIED: Fifty-four client-owned dogs. PROCEDURES: A multicenter, retrospective study was performed including ECS with suspected PRA which underwent phacoemulsification. PRA was suspected on ophthalmic examination before and after surgery, and/or after electroretinography (ERG) was performed. Visual outcome was assessed by menace response per eye at seven time periods post-surgery (P1= 25-90 days, P2 = 91-180 days, P3 = 181-364 days, P4 = 365-549 days, P5 = 550-729 days, P6 = 730-1094 days, and P7 ≥ 1095 days). Descriptive statistics were performed. Generalized estimating equations were used to identify predictors associated with vision after surgery. Odds ratio and confidence intervals were reported. Significance was set at P < 0.05. Owners were invited to participate in a questionnaire. RESULTS: Phacoemulsification was performed in 85 eyes. Median age at surgery was 9.09 years (min. 2.17 years, max. 13.49 years). At all re-examinations, up to and including P5, significantly more eyes were visual than before surgery (P ≤ 0.003). Odds for vision were significantly increased for eyes that underwent surgery. Electroretinograms were performed in 75/85 eyes that underwent surgery, demonstrating low b-wave amplitudes. There was no significant effect of the age, gender, vision before surgery, presence of dazzle reflex, cataract stage or abnormality on gonioscopy on visual outcome. The questionnaire response rate was 48.2%. Most participants (92.5%) felt that cataract surgery led to improvement of the dog's quality of life. CONCLUSIONS: English Cocker Spaniels with suspected PRA may benefit from phacoemulsification with vision up to 2 years following surgery.

Immunohistochemical characterization of feline lymphoplasmacytic anterior uveitis.

OBJECTIVE: To characterize the immune cells present in different forms of feline anterior uveitis. SAMPLES: Eyes were obtained from 49 cats diagnosed with chronic idiopathic lymphoplasmacytic anterior uveitis, 7 cats with feline infectious peritonitis (FIP), and 9 cats euthanized for nonocular disease. METHODS: H&E sections were scored on the level of infiltrate in the anterior uvea. Immunohistochemistry was performed for FoxP3, CD3, and IL-17A, and positive cells were quantified in multiple images of each sample. A generalized estimating equation tested for an association between the level of inflammation and the prevalence of these cell types. RESULTS: Cells stained positive for IL-17A in idiopathic uveitis but not in FIP samples. We found significantly fewer FoxP3+ and CD3+ cells in low-grade compared with high-grade inflammation in idiopathic uveitis or FIP samples (P values all <.005), but no difference between FIP and high-grade samples. CONCLUSIONS: Idiopathic, but not FIP-associated, uveitis appears to have Th17 cell involvement. The numbers of FoxP3+ and CD3+ T-cells present appear directly correlated; thus, the severity of disease does not appear directly determined by the numbers of regulatory cells.

Prevalence of Avian Influenza A(H5) and A(H9) Viruses in Live Bird Markets, Bangladesh.

We conducted a cross-sectional study in live bird markets (LBMs) in Dhaka and Chittagong, Bangladesh, to estimate the prevalence of avian influenza A(H5) and A(H9) viruses in different types of poultry and environmental areas by using Bayesian hierarchical logistic regression models. We detected these viruses in nearly all LBMs. Prevalence of A(H5) virus was higher in waterfowl than in chickens, whereas prevalence of A(H9) virus was higher in chickens than in waterfowl and, among chicken types, in industrial broilers than in cross-breeds and indigenous breeds. LBMs with >1 wholesaler were more frequently contaminated by A(H5) virus than retail-only LBMs. Prevalence of A(H9) virus in poultry and level of environmental contamination were also higher in LBMs with >1 wholesaler. We found a high level of circulation of both avian influenza viruses in surveyed LBMs. Prevalence was influenced by type of poultry, environmental site, and trading.

Evaluation of the dynamic predictors of fluid responsiveness in dogs receiving goal-directed fluid therapy.

OBJECTIVES: Goal-directed fluid therapy (GDFT) based on pulse pressure variation (PPV) was used in anaesthetized dogs undergoing abdominal surgeries. The aims were 1) to evaluate the success rate of the PPV ≥13% in detecting fluid responsiveness [delta stroke volume (ΔSV) ≥10%]; 2) to assess the correlation between PPV, systolic pressure variation (SPV), Plethysmograph Variability Index (PVI) and central venous pressure (CVP) and 3) to establish the threshold value for the PVI that would predict a PPV value of ≥13% and indirectly discriminate responders from nonresponders to fluid therapy. STUDY DESIGN: Clinical, prospective, interventional study. ANIMALS: A total of 63 client-owned dogs scheduled for abdominal procedures. METHODS: PPV and SPV were calculated manually from the invasive blood pressure trace on the Datex monitor. PVI was recorded from the Masimo pulse oximeter. Fluid challenge (10 mL kg-1 Compound Sodium Lactate) was performed when PPV was ≥13% and/or mean arterial pressure (MAP) < 60 mmHg. Fluid responsiveness was assessed by the transoesophageal Doppler probe. Cardiovascular parameters (heart rate, MAP, PPV, SPV, PVI, SV and if available, CVP) were measured before and after each fluid intervention. RESULTS: PPV ≥ 13% reliably predicted fluid responsiveness in 82.9% of cases. There was positive correlation between PPV and SPV (r = 0.84%), PPV and logPVI (r = 0.46) as well as SPV and logPVI (r = 0.45). Noninvasive PVI value ≥13% should predict PPV threshold value (13%) with 97% sensitivity and 68% specificity. There was no statistically significant correlation between PPV and CVP. CONCLUSIONS: PPV is a useful clinical tool to detect occult hypovolaemia and predict cardiovascular response to fluid challenge. Use of PPV is recommended as a part of GDFT in dogs undergoing abdominal procedures.

Development and initial validation of a sensory threshold examination protocol (STEP) for phenotyping canine pain syndromes.

OBJECTIVE: To study the feasibility and test-retest repeatability of a sensory threshold examination protocol (STEP) and report the quantitative sensory threshold distributions in healthy dogs. STUDY DESIGN: Prospective, observational, cohort study. ANIMALS: Twenty-five healthy client-owned dogs. METHODS: Tactile sensitivity test (TST) (von Frey filaments), mechanical thresholds (MT with 2, 4 and 8 mm probes), heat thresholds (HT) and responsiveness to cold stimulus (CT at 0 °C) were quantitatively assessed for five body areas (BAs; tibias, humeri, neck, thoracolumbar region and abdomen) in a randomized order on three different occasions. Linear mixed model and generalized linear mixed models were used to evaluate the effects of body weight category, age, sex, BA, occasion, feasibility score and investigator experience. Test-retest repeatability was evaluated with the intra-class correlation coefficient. RESULTS: The STEP lasted 90 minutes without side effects. The BA affected most tests (p ≤ 0.001). Higher thresholds and longer cold latencies were scored in the neck (p ≤ 0.024) compared to other BAs. Weight category affected all thresholds (p ≤ 0.037). Small dogs had lower MT (∼1.4 N mean difference) and HT (1.1 °C mean difference) than other dogs (p ≤ 0.029). Young dogs had higher HT than adults (2.2 °C mean difference) (p = 0.035). Gender also affected TST, MT and HT (p < 0.05) (females versus males: TST odds ratio = 0.5, MT = 1.3 N mean difference, HT = 2.2 °C mean difference). Repeatability was substantial to moderate for all tests, but poor for TST. There was no difference in thresholds between occasions, except for CT. Test-retest repeatability was slightly better with the 2 mm MT probe compared to other diameters and improved with operator experience. CONCLUSIONS AND CLINICAL RELEVANCE: The STEP was feasible, was well tolerated and showed substantial test-retest repeatability in healthy dogs. Further validation is needed in dogs suffering pain.

A pilot study of the effects of acupuncture treatment on objective and subjective gait parameters in horses.

OBJECTIVES: To investigate whether acupuncture can alter gait in horses as assessed by objective and subjective parameters. STUDY DESIGN: Prospective, randomized, singleblinded, crossover study. ANIMALS: Eight adult horses. METHODS: Horses were randomly assigned to a treatment (three acupuncture treatments in 8 days) or control group. Subjective and objective gait analyses were performed before and after each treatment and at 1, 3 and 7 days after the last treatment (time-points 1-9, respectively). Horses were assessed at the trot in a straight line on a hard surface and on the lunge on the left and right reins on a soft surface (conditions 1-3, respectively). After 12 weeks, groups were reversed. Objective gait analysis was performed using inertial sensors and subjective analysis by two board- certified surgeons who reviewed video-recordings. Each limb was assessed for lameness before and after treatment. Lameness and global scores were assigned using 4-point scales. Assessors were blinded to treatment status. The effects of treatment (yes/no), time (1-9) and horse under conditions 1 -3 were compared using a linear mixed-effects model and a generalized estimating equation. RESULTS: Treatment decreased hip hike difference under all conditions [condition 1: control, 6.3 ± 6.4 mm versus treatment, -0.2 ± 6.4 mm (p = 0.007); condition 2: control, 9.7 ± 7.8 mm versus treatment, 2.8 ± 7.8 mm (p = 0.032); condition 3: control, 7.3 ± 6.3 mm versus treatment, -2.7 ± 6.4 mm (p = 0.003)]. Other parameters also improved significantly under conditions 1 and 3. Based on subjective gait analysis, treatment decreased lameness [odds ratio (OR) 0.51, 95% confidence interval (CI) 0.34-0.78; p = 0.002] but not global (OR 0.53, 95% CI 0.24-1.10; p = 0.12) scores. CONCLUSIONS AND CLINICAL RELEVANCE: Acupuncture can change horses' gaits to a degree appreciable by objective and subjective analyses.

Interaction of chlorhexidine with trisEDTA or miconazole in vitro against canine meticillin-resistant and -susceptible Staphylococcus pseudintermedius isolates from two UK regions.

BACKGROUND: Topical therapy is an important alternative to systemic antibacterial therapy for treatment of canine superficial pyoderma in light of the emergence of multidrug-resistant staphylococci. Chlorhexidine is widely used in shampoo products alone or in combination with miconazole or tromethamine-ethylenediaminetetraacetic acid (trisEDTA). Comparisons of these combinations have not been made. HYPOTHESIS/OBJECTIVES: To determine minimum inhibitory concentrations (MICs) of combinations of chlorhexidine/miconazole and chlorhexidine/trisEDTA in vitro in a collection of Staphylococcus pseudintermedius (SP) from northern (NUK) and southeastern (SEUK) United Kingdom (UK) sources. METHODS: MICs of chlorhexidine, miconazole, trisEDTA and combinations of chlorhexidine/miconazole (1:1) or chlorhexidine/trisEDTA (80:16:1 and 80:5:1) were determined for 196 canine SP isolates from NUK [49 meticillin-resistant (MRSP), 50 meticillin-susceptible (MSSP)] and fom SEUK (48 MRSP, 49 MSSP) using agar dilution. RESULTS: TrisEDTA alone did not inhibit growth. Chlorhexidine/miconazole MICs (median = 0.5 mg/L) were lower than those of either drug alone (P < 0.05) and lower than chlorhexidine/trisEDTA MICs (median = 1 mg/L; P < 0.0005) in each bacterial type and from both regions, except for miconazole in NUK MSSP. An additive interaction was noted between chlorhexidine and miconazole or trisEDTA (80:16:1 ratio) in 79 and 43 isolates, respectively, whereas antagonism between chlorhexidine and trisEDTA was noted for three isolates. NUK isolates were more susceptible than SEUK isolates (P < 0.05), except MRSP exposed to chlorhexidine and the chlorhexidine/trisEDTA (80:16:1) combination. CONCLUSIONS AND CLINICAL IMPORTANCE: These low MICs are likely to be exceeded by topical therapy. Evaluation of the mechanisms by which chlorhexidine combinations interact to reduce MICs is warranted, in view of increasing concerns of biocide tolerance in staphylococci.

Extracorporeal liver assist device to exchange albumin and remove endotoxin in acute liver failure: Results of a pivotal pre-clinical study.

BACKGROUND & AIMS: In acute liver failure, severity of liver injury and clinical progression of disease are in part consequent upon activation of the innate immune system. Endotoxaemia contributes to innate immune system activation and the detoxifying function of albumin, critical to recovery from liver injury, is irreversibly destroyed in acute liver failure. University College London-Liver Dialysis Device is a novel artificial extracorporeal liver assist device, which is used with albumin infusion, to achieve removal and replacement of dysfunctional albumin and reduction in endotoxaemia. We aimed to test the effect of this device on survival in a pig model of acetaminophen-induced acute liver failure. METHODS: Pigs were randomised to three groups: Acetaminophen plus University College London-Liver Dialysis Device (n=9); Acetaminophen plus Control Device (n=7); and Control plus Control Device (n=4). Device treatment was initiated two h after onset of irreversible acute liver failure. RESULTS: The Liver Dialysis Device resulted in 67% reduced risk of death in acetaminophen-induced acute liver failure compared to Control Device (hazard ratio=0.33, p=0.0439). This was associated with 27% decrease in circulating irreversibly oxidised human non-mercaptalbumin-2 throughout treatment (p=0.046); 54% reduction in overall severity of endotoxaemia (p=0.024); delay in development of vasoplegia and acute lung injury; and delay in systemic activation of the TLR4 signalling pathway. Liver Dialysis Device-associated adverse clinical effects were not seen. CONCLUSIONS: The survival benefit and lack of adverse effects would support clinical trials of University College London-Liver Dialysis Device in acute liver failure patients.