Relative egg extraction efficiencies of manual and automated fecal egg count methods in equines.

The World Association for the Advancement of Veterinary Parasitology recently released new recommendations for the design of fecal egg count (FEC) reduction tests for livestock. These provide suggestions as to the number of animals to be sampled and the minimum number of eggs that must be counted to produce statistically meaningful results. One of the considerations for study design is the multiplication factor of the FEC method to be used; methods with lower multiplication factors require fewer animals to be sampled because they are presumed to count more eggs per test. However, multiplication factor is not the sole determinant of the number of eggs counted by any given method, since different techniques use very different sample extraction methodologies that could affect the number of eggs detected beyond just the amount of feces examined. In this light, we compared three commonly used manual FEC methods (mini-FLOTAC, McMaster and Wisconsin) and two automated methods (Imagyst and Parasight All-in-One) with respect to how many equine strongylid and ascarid eggs they counted in the same samples. McMaster and mini-FLOTAC (multiplication factors of 25x and 5x, respectively) produced the most accurate results of the methods tested but mini-FLOTAC counted approximately 5-times more eggs than McMaster. However, Wisconsin and Parasight (multiplication factor = 1x) counted 3-times more ova than mini-FLOTAC, which was less than the 5-fold difference in their multiplication factors. As a result, these tests perform with multiplication factors more akin to 1.6x relative to mini-FLOTAC. Imagyst, due to its unique sample preparation methodology, does not have a traditional multiplication factor but performed similarly to McMaster with respect to egg recovery.

The role of macrophages in the development of biliary injury in a lipopolysaccharide-aggravated hepatic ischaemia-reperfusion model.

INTRODUCTION: Endotoxins, in the form of lipopolysaccharides (LPS), are potent inducers of biliary injury. However the mechanism by which injury develops remains unclear. We hypothesized that hepatic macrophages are pivotal in the development of endotoxin-induced biliary injury and that no injury would occur in their absence. MATERIAL AND METHODS: Clodronate liposomes were used to deplete macrophages from the liver. Forty-eight rats were equally divided across six study groups: sham operation (sham), liposome treatment and sham operation (liposomes+sham), 1mg/kg LPS i.p. (LPS), liposome treatment and LPS administration (liposomes+LPS), hepatic ischaemia-reperfusion injury with LPS administration (IRI+LPS) and liposome treatment followed by IRI+LPS (liposomes+IRI+LPS). Following 6h of reperfusion, blood, bile, and liver tissue was collected for further analysis. Small bile duct injury was assessed, serum liver tests were performed and bile composition was evaluated. The permeability of the blood-biliary barrier (BBB) was assessed using intravenously administered horseradish peroxidase (HRP). RESULTS: The presence of hepatic macrophages was reduced by 90% in LPS and IRI+LPS groups pre-treated with clodronate liposomes (P<0.001). Severe small bile duct injury was not affected by macrophage depletion, and persisted in the liposomes+IRI+LPS group (50% of animals) and liposomes+LPS group (75% of animals). Likewise, BBB impairment persisted following macrophage depletion. LPS-induced elevation of the chemokine Mcp-1 in bile was not affected by macrophage depletion. CONCLUSIONS: Depletion of hepatic macrophages did not prevent development of biliary injury following LPS or LPS-enhanced IRI. Cholangiocyte activation rather than macrophage activation may underlie this injury. This article is part of a Special Issue entitled: Cholangiocytes in Health and Diseaseedited by Jesus Banales, Marco Marzioni, Nicholas LaRusso and Peter Jansen.

Invasive melanoma in vivo can be distinguished from basal cell carcinoma, benign naevi and healthy skin by canine olfaction: a proof-of-principle study of differential volatile organic compound emission.

BACKGROUND: Volatile organic compounds (VOCs) are continuously released by the body during normal metabolic processes, but their profiles change in the presence of cancer. Robust evidence that invasive melanoma in vivo emits a characteristic VOC signature is lacking. OBJECTIVES: To conduct a canine olfactory, proof-of-principle study to investigate whether VOCs from invasive melanoma are distinguishable from those of basal cell carcinoma (BCC), benign naevi and healthy skin in vivo. METHODS: After a 13-month training period, the dog's ability to discriminate melanoma was evaluated in 20 double-blind tests, each requiring selection of one melanoma sample from nine controls (three each of BCC, naevi and healthy skin; all samples new to the dog). RESULTS: The dog correctly selected the melanoma sample on nine (45%) occasions (95% confidence interval 0·23-0·68) vs. 10% expected by chance alone. A one-sided exact binomial test gave a P-value of < 0·01, supporting the hypothesis that samples were not chosen at random but that some degree of VOC signal from the melanoma samples significantly increased the probability of their detection. Use of a discrete-choice model confirmed melanoma as the most influential of the recorded medical/personal covariates in determining the dog's choice of sample. Accuracy rates based on familiar samples during training were not a reliable indicator of the dog's ability to distinguish melanoma, when confronted with new, unknown samples. CONCLUSIONS: Invasive melanoma in vivo releases odorous VOCs distinct from those of BCC, benign naevi and healthy skin, adding to the evidence that the volatile metabolome of melanoma contains diagnostically useful biomarkers.

Heart disease management by women: does intervention format matter?

A randomized controlled trial of two formats of a program (Women Take PRIDE) to enhance management of heart disease by patients was conducted. Older women (N = 575) were randomly assigned to a group or self-directed format or to a control group. Data regarding symptoms, functional health status, and weight were collected at baseline and at 4, 12, and 18 months. The formats produced different outcomes. At 18 months, the self-directed format was better than the control in reducing the number (p ≤ .02), frequency (p ≤ .03), and bothersomeness (p ≤ .02) of cardiac symptoms. The self-directed format was also better than the group format in reducing symptom frequency of all types (p ≤ .04). The group format improved ambulation at 12 months (p ≤ .04) and weight loss at 18 months (p ≤ .03), and group participants were more likely to complete the program (p ≤ .05). The availability of different learning formats could enhance management of cardiovascular disease by patients.

Visitor assessment of the mandatory alternative transportation system at Zion National Park.

Transportation infrastructure in national parks has historically been designed for the automobile. With more vehicles in the parks, visitors found themselves in circumstances more reminiscent of a city than a park. Traffic jams, overcrowding, illegal parking, horn honking, and idling vehicles became common, creating stress and contributing to air and noise pollution, the very things visitors were hoping to get away from. Park managers began searching for alternatives, including shuttle systems. Many national parks have implemented optional shuttle systems, but relatively few have completely closed roads to vehicles, transporting visitors on mandatory shuttles. Zion National Park instituted a mandatory shuttle system in May 2000 to relieve crowding and congestion in the main canyon and to protect natural resources. Taking a longitudinal approach, attributes of the shuttle (e.g., crowding, accessibility, freedom, efficiency, preference, and success) were assessed with experiential park factors (e.g., scenic beauty, naturalness, solitude, tranquility, air quality, and soundscape) in 2000, 2003, and 2010 by surveying shuttle-riding park visitors. While visitors initially reported a few reservations about the shuttle system, by 2003, the majority rated the system successful. Ratings of all shuttle-related variables, except crowding, improved over the decade. Improvements were greatest for freedom, accessibility, and efficiency. Multiple regression found overall shuttle success to be mediated by preference, freedom, accessibility, efficiency, and comfort. Experiential variables assessing park conditions followed a similar pattern, with improved ratings as the decade progressed. Results provide important insights into the visitor experience with mandatory alternative shuttle systems in national parks.

Osteonecrosis in children after allogeneic hematopoietic cell transplantation: study of prevalence, risk factors and longitudinal changes using MR imaging.

Osteonecrosis after hematopoietic SCT (HCT) has seldom been addressed in pediatric populations. At our institution, since January 2002, children undergoing allogeneic HCT (alloHCT) receive yearly follow-up magnetic resonance imaging (MR) of hips and knees. To estimate the prevalence, longitudinal changes and associated risk factors for osteonecrosis after alloHCT, we reviewed MRs for children who underwent single alloHCT during the study period. We analyzed 149 of 344 patients who had post-HCT MR imaging performed (84 males; median age 11 years (range, 0.5-21 years)), median follow-up time was 32.6 months (range, 2.8-97.2 months). In all, 44 (29.5%) developed osteonecrosis of hips and/or knees; of those, 20 (45%) had at least 30% epiphyseal involvement. In 23 (52%), osteonecrosis lesions were identified in the first and in 43 (98%) by the third yearly scan. Knees were more frequently involved than hips; severity of osteonecrosis was greater in hips. Those who had pre-alloHCT osteonecrosis, two patients' hips and six patients' knees resolved completely; three patients' osteonecrosis lesions regressed after alloHCT. On risk factor analysis, age at time of alloHCT (P=0.051) and osteonecrosis identified by MRs before alloHCT (P=0.001) were the primary risk factors. This analysis shows that preventive strategies for osteonecrosis in this population should focus on measures to minimize risk factors before alloHCT.

Serum biomarkers of aging in the Brown Norway rat.

Serum biomarkers to identify susceptibility to disease in aged humans are well researched. On the other hand, our understanding of biomarkers in animal models of aging is limited. Hence, we applied a commercially available panel of 58 serum analytes to screen for possible biomarkers of aging in 4, 12, and 24 month old Brown Norway rats. We found that serum levels of 5 of the 58 analytes were significantly affected by age: C-reactive protein (CRP), myoglobin, macrophage derived chemokine-2 (MDC), fibroblast growth factor-basic, and vascular cell adhesion molecule-1. Among these analytes, CRP was the only one that increased with aging. The variability of CRP and MDC-2 was relatively low compared to the other analytes of the panel. It is concluded that CRP and possibly MDC-2 are candidates for biomarkers of aging in the BN rat.

Prevalence of osteonecrosis and associated risk factors in children before allogeneic BMT.

Osteonecrosis (ON) is a debilitating long-term complication of allogeneic BMT (allo-BMT), but may begin before allo-BMT in some children because of their primary disease treatment. Therefore, to estimate the prevalence and associated risk factors for ON before allo-BMT, we conducted a retrospective analysis of magnetic resonance (MR) studies of 118 children who underwent first allo-BMT at our institution between December 2000 and September 2007. Of the 118 consecutive patients, 107 (90.7%) underwent prospective MR studies irrespective of symptoms (69 males; median age at allo-BMT 12.9 years), and 11 underwent MR studies for symptoms. Among the 107 who had prospective imaging, 23 (21.5%) had ON; nearly 50% had at least 30% epiphyseal involvement. Knees were more frequently involved than were hips; severity of ON was greater in hips. ON prevalence before allo-BMT was 23.72% when all 118 patients were included in the denominator. Risk factor analysis, limited to MR studies performed irrespective of symptoms, revealed female gender (P=0.049) and age 10 years at the time of MR study (P=0.03) as significant risk factors, and primary diagnosis of lymphoid malignancies and aplastic anemia trended toward significance. ON before allo-BMT is a common occurrence in children.

Mycophenolate mofetil treatment in dogs with serologically diagnosed acquired myasthenia gravis: 27 cases (1999-2008).

OBJECTIVE-To compare clinical outcome in dogs with serologically diagnosed acquired myasthenia gravis (MG) treated with pyridostigmine bromide (PYR) with that of dogs treated with mycophenolate mofetil (MMF) and PYR (MMF + PYR). DESIGN-Retrospective case series. ANIMALS-27 dogs. PROCEDURES-Medical records from August 1999 through February 2008 were reviewed to identify dogs with serologically diagnosed acquired MG treated with PYR or MMF + PYR. Data collected for each dog included signalment, whether the dog had megaesophagus or pneumonia (or both), thyroid hormone concentration, remission, time to remission, and survival time. Rates for detection of clinical signs and survival time were compared. Survival time was estimated via the Kaplan-Meier method. Influence of drug treatment protocol on likelihood of remission, time to remission, and survival time was examined. Effects of MMF treatment, megaesophagus, pneumonia, and low serum thyroid hormone concentration on time to remission and survival time were also analyzed. RESULTS-12 dogs were treated with PYR, and 15 were treated with MMF + PYR. Mortality rates were 33% (PYR) and 40% (MMF + PYR). There was pharmacological remission in 5 and 6 dogs in the PYR and MMF + PYR groups, respectively. No significant differences were detected between treatment groups for remission rate, time to remission, or survival time. Megaesophagus, pneumonia, and low serum thyroid hormone concentration had no significant effect on time to remission or survival time for either treatment group. CONCLUSIONS AND CLINICAL RELEVANCE-The results did not support routine use of MMF for the treatment of dogs with acquired MG.

Pregabalin as an adjunct to phenobarbital, potassium bromide, or a combination of phenobarbital and potassium bromide for treatment of dogs with suspected idiopathic epilepsy.

OBJECTIVE: To assess tolerability and short-term efficacy of oral administration of pregabalin as an adjunct to phenobarbital, potassium bromide, or a combination of phenobarbital and potassium bromide for treatment of dogs with poorly controlled suspected idiopathic epilepsy. DESIGN: Open-label, noncomparative clinical trial. ANIMALS: 11 client-owned dogs suspected of having idiopathic epilepsy that was inadequately controlled with phenobarbital, potassium bromide, or a combination of these 2 drugs. PROCEDURES: Dogs were treated with pregabalin (3 to 4 mg/kg [1.4 to 1.8 mg/lb], PO, q 8 h) for 3 months. Number of generalized seizures in the 3 months before and after initiation of pregabalin treatment was recorded. Number of responders (>or= 50% reduction in seizure frequency) was recorded, and seizure frequency before and after initiation of pregabalin treatment was compared by use of a nonparametric Wilcoxon signed rank test. RESULTS: Seizures were significantly reduced (mean, 57%; median, 50%) after pregabalin administration in the 9 dogs that completed the study; 7 were considered responders with mean and median seizure reductions of 64% and 58%, respectively. Adverse effects for pregabalin were reported in 10 dogs. Mean and median plasma pregabalin concentrations for all dogs were 6.4 and 7.3 microg/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Pregabalin may hold promise as a safe and effective adjunct anticonvulsant drug for epileptic dogs poorly controlled with the standard drugs phenobarbital or potassium bromide. Adverse effects of pregabalin appeared to be mild. Additional studies with larger numbers of dogs and longer follow-up intervals are warranted.

Pharmacokinetics of single-dose oral pregabalin administration in normal dogs.

OBJECTIVE: To describe the pharmacokinetics of pregabalin in normal dogs after a single oral dose. STUDY DESIGN: Prospective experiment. ANIMALS: Six adult Labrador/Greyhound dogs (four females and two males) aged 2.6 (2.6-5.6) years old (median and range) weighing 33.4 (26.8-42.1) kg. METHODS: After jugular vein catheterization, the dogs received a single oral dose of pregabalin ( approximately 4 mg kg(-1)). Blood samples were collected at: 0 (before drug administration), 15 and 30 minutes and at 1, 1.5, 2, 3, 4, 6, 8, 12, 24 and 36 hours after drug administration. Plasma pregabalin concentration was measured by HPLC. Noncompartmental analysis was used to estimate pharmacokinetic variables. RESULTS: No adverse effects were observed. The median (range) pharmacokinetic parameters were: Area under the curve from time 0 to 36 hours = 81.8 (56.5-92.1) microg hour mL(-1); absorption half-life = 0.38 (0.25-1.11) hours; elimination half-life = 6.90 (6.21-7.40) hours; time over 2.8 microg mL(-1) (the presumed minimal effective concentration) = 11.11 (6.97-14.47) hours; maximal plasma concentration (C(max)) = 7.15 (4.6-7.9) microg mL(-1); time for C(max) to occur = 1.5 (1.0-4.0) hours. Assuming an 8-hour dosing interval, predicted minimal, average, and maximal steady state plasma concentrations were 6.5 (4.8-8.1), 8.8 (7.3-10.9), and 13.0 (8.8-15.2) microg mL(-1). The corresponding values assuming a 12-hour interval were 3.8 (2.4-4.8), 6.8 (4.9-7.9), and 10.1 (6.6-11.6) microg mL(-1). CONCLUSIONS AND CLINICAL RELEVANCE: Pregabalin 4 mg kg(-1) PO produces plasma concentrations within the extrapolated therapeutic range from humans for sufficient time to suggest that a twice daily dosing regime would be adequate. Further study of the drug's safety and efficacy for the treatment of neuropathic pain and seizures in dogs is warranted.

Association of marijuana use and the incidence of testicular germ cell tumors.

BACKGROUND: The incidence of testicular germ cell tumors (TGCTs) has been increasing the past 4 to 6 decades; however, exposures that account for this rise have not been identified. Marijuana use also grew during the same period, and it has been established that chronic marijuana use produces adverse effects on the human endocrine and reproductive systems. In this study, the authors tested the hypothesis that marijuana use is a risk factor for TGCT. METHODS: A population-based, case-control study of 369 men ages 18 to 44 years who were diagnosed with TGCT from January 1999 through January 2006 was conducted in King, Pierce and Snohomish Counties in Washington State. The responses of these men to questions on their lifetime marijuana use were compared with the responses of 979 age-matched controls who resided in the same 3 counties during the case diagnosis period. RESULTS: Men with a TGCT were more likely to be current marijuana smokers at the reference date compared with controls (odds ratio [OR], 1.7; 95% confidence interval [95% CI], 1.1-2.5). In analyses according to histologic type, most of the association between current marijuana use and TGCT was observed in men who had nonseminomas/mixed histology tumors (current use: OR, 2.3; 95% CI, 1.3-4.0). Age at first use among current users (age<18 years [OR, 2.8] vs age>or=18 years [OR, 1.3]) and frequency of use (daily or weekly [OR, 3.0] vs less than once per week [OR, 1.8]) appeared to modify the risk. CONCLUSIONS: An association was observed between marijuana use and the occurrence of nonseminoma TGCTs. Additional studies of TGCTs will be needed to test this hypothesis, including molecular analyses of cannabinoid receptors and endocannabinoid signaling, which may provide clues regarding the biologic mechanisms of TGCTs.

Heart disease management by women: does intervention format matter?

A randomized controlled trial of two formats of a program (Women Take PRIDE) to enhance management of heart disease by patients was conducted. Older women (N = 575) were randomly assigned to a group or self-directed format or to a control group. Data regarding symptoms, functional health status, and weight were collected at baseline and at 4, 12, and 18 months. The formats produced different outcomes. At 18 months, the self-directed format was better than the control in reducing the number (p < or = .02), frequency (p < or = .03), and bothersomeness (p < or = .02) of cardiac symptoms. The self-directed format was also better than the group format in reducing symptom frequency of all types (p < or = .04). The group format improved ambulation at 12 months (p < or = .04) and weight loss at 18 months (p < or = .03), and group participants were more likely to complete the program ( p < or = .05). The availability of different learning formats could enhance management of cardiovascular disease by patients.

Effects of adenosine infusion on the minimum alveolar concentration of isoflurane in dogs.

OBJECTIVE: To determine the effects of adenosine infusion on the minimum alveolar concentration (MAC) of isoflurane in dogs. STUDY DESIGN: Prospective, randomized crossover study. ANIMALS: Seven adult male and female Beagles weighing 10.9 (7.5, 13.6) kg [median (minimum, maximum)]. METHODS: Each dog was anesthetized with isoflurane in oxygen and randomly assigned to receive either an intravenous (IV) adenosine (0.3 mg kg(-1) minute(-1)) or saline (6 mL kg(-1) hour(-1) IV) infusion. After an interval of 7 days or more, each dog was re-anesthetized and treated with the alternative infusion. Using a tail-clamp technique, MAC was determined before (pre-infusion), during (infusion), and 2 hours after the infusions (post-infusion). RESULTS: The pre-infusion MAC of isoflurane was 1.25 (1.15, 1.35) [median (minimum, maximum)] vol.% for the saline treatment group and 1.25 (1.05, 1.45) vol.% for the adenosine treatment group, and did not differ significantly between the two treatments. The infusion MAC values were not significantly different (p = 0.16) and were 1.25 (0.95, 1.35) vol.% and 1.05 (1.00, 1.25) vol.%, respectively. The post-infusion MAC values differed significantly (p = 0.016); MAC was 1.15 (1.15, 1.35) vol.% and 1.05 (1.05, 1.25) vol.% for the saline and adenosine treatment groups, respectively. During infusion, mean arterial blood pressure decreased significantly (p = 0.008) during adenosine treatment compared with the saline 66 mmHg (52, 72) and 91 mmHg (68, 110), respectively. End-tidal CO2 (Pe'CO2), urine production, hematocrit, and plasma total solids did not differ significantly between the two treatments at any time (all p > 0.05). CONCLUSION: Although the MAC of isoflurane in dogs was not decreased significantly during infusion with adenosine (0.3 mg kg(-1) minute(-1)), it was significantly decreased post-infusion, but only by 0.1 vol.%, an amount not considered clinically important. Adenosine infusion decreased mean arterial pressure by 27% and did not adversely affect renal function.

Reduced levels of glutathione S-transferases in patch test reactions to dithranol and sodium lauryl sulphate as demonstrated by quantitative immunocytochemistry: evidence for oxidative stress in acute irritant contact dermatitis.

There is increasing evidence that oxidative stress plays a role in the pathogenesis of acute irritant contact dermatitis. As part of on-going studies into the effect of irritant chemicals on the anti-oxidant enzyme systems in the skin, we have examined the changing levels of two classes of glutathione S-transferase in patch test reactions to dithranol and sodium lauryl sulphate, using quantitative immunocytochemistry. Although no changes were evident after 6 hrs, significant reductions in the density of staining for glutathione S-transferase alpha were seen with both irritants after 48 hrs and 96 hrs. Glutathione S-transferase pi levels were reduced to a lesser degree, reaching significance for dithranol at the 96 hrs time point only, and for sodium lauryl sulphate at 48 hrs only. The results support the hypothesis that oxidative stress plays a role in chemically-induced inflammation, not only in the case of irritants such as dithranol which are known to directly generate reactive oxygen species, but also with chemicals not generally associated with free radical generation.

A review of "noncontingent" reinforcement as treatment for the aberrant behavior of individuals with developmental disabilities.

The term noncontingent reinforcement (NCR) refers to the delivery of an aberrant behavior's known reinforcer on a response-independent basis. The typical result is a decrease in responding from baseline (i.e., reinforcement) levels. NCR has become one of the most reported function-based treatments for aberrant behavior in the recent literature. The purpose of this review is to briefly discuss the history of the procedure and summarize the findings from the treatment research literature. The review is organized into the following sections: (a) basic research on NCR, (b) NCR as a control procedure, (c) NCR as a function-based treatment, (d) considerations in the programming of NCR schedules, (e) behavior-change mechanisms underlying NCR effects, and (t) directions for future research.

A variation of noncontingent reinforcement in the treatment of aberrant behavior.

We examined the effectiveness of a variation of noncontingent reinforcement (NCR) that incorporated a stimulus-delay procedure in the reduction of aberrant behavior maintained by positive reinforcement. Functional analyses for three individuals diagnosed with developmental disabilities indicated that their behaviors were maintained by positive reinforcement: one in the form of access to a tangible item, another by attention, and the third by physical contact. We implemented NCR with the delay procedure with two participants using reversal designs to evaluate effects. We also compared this NCR variation and DRO with the third participant to evaluate reinforcer-delivery rates. The variation of NCR was successful in reducing all aberrant behavior to near-zero levels. A comparison of reinforcer delivery between NCR with the stimulus-delay procedure and DRO demonstrated that the participant accessed more reinforcement with NCR. Results are discussed in the context of enhancing decelerative interventions with emphases on minimizing response effort for caregivers and maximizing access to reinforcement for the individuals.