Systematic evaluation supports the use of ELISA for quantification of calprotectin in equine feces, a first step toward noninvasive quantification of intestinal inflammation in horses.

OBJECTIVE: To optimize and evaluate methods for the detection of the inflammatory biomarkers myeloperoxidase (MPO) and calprotectin (CP) in equine feces by ELISA. ANIMALS: Healthy horses (n = 28) and horses with intestinal inflammation (n = 10). METHODS: Feces were suspended in buffer to create fecal supernatant. Serum and fecal supernatant were analyzed using ELISA kits validated for the detection of MPO and CP in equine serum. Assay validation steps included intra- and interassay variability (coefficient of variation [CV]), dilution linearity, spike recovery, and sample type correlation. Variations in sample handling protocols (centrifugation speed, extraction buffer, and filtration) were evaluated. RESULTS: 17 paired fecal and serum samples were used for initial analysis (10 healthy horses, 7 colitis). Previously reported sample handling protocols resulted in detectable MPO and CP but poor CV, linearity, and spike recovery. There was a linear correlation between serum and fecal samples for CP but not MPO. There was a significant difference between the concentration and CV of alternative sample handling protocols for CP and MPO, with improved CV for CP (2.1% to 18.6%) but not MPO (14.4% to 53.4%). Processing fresh feces with a fecal extraction buffer and filtration of supernatant resulted in the best CV (0.5% to 3.8%) and recovery (45% to 64%) for CP. Detection of MPO was inconsistent regardless of method. CLINICAL RELEVANCE: There are few reliable diagnostic modalities for inflammation of the equine large colon. Findings support quantification of CP in equine feces using the described ELISA kit and protocol. With additional study to establish reference interval and clinical utility, the fecal inflammatory biomarker CP may allow for noninvasive quantification of intestinal inflammation in horses.

Acute phase protein concentrations following serial procaine penicillin G injections in horses.

BACKGROUND: Acute phase protein (APP) measurement is used to detect inflammation. Intramuscular (IM) injections could cause tissue injury and induce an acute phase response (APR). OBJECTIVES: To evaluate the effects of IM procaine penicillin G (PPG) injections on APP concentrations in horses. STUDY DESIGN: Prospective longitudinal design. METHODS: PPG was administered intramuscularly to six horses, twice daily, for 5 days. Plasma fibrinogen (FIB), serum amyloid A (SAA), haptoglobin (HAP), creatine kinase (CK), and aspartate aminotransferase (AST) were quantified daily for 5 days before the first injection, during the course of administration, and for 4 days after the final dose. Analytes were quantified every other day for the remaining 16 days. Data were compared using a parametric or non-parametric repeated measures ANOVA and a Tukey's or Mann-Whitney rank sum test, respectively. Significance was set at p < 0.05. RESULTS: CK was increased over baseline (mean ± SD: 200 ± 74 IU/L) on Days 1-6 (p < 0.001 to p = 0.02, mean ± SD: 723-1177 ± 355-544 IU/L) and AST was increased above baseline (mean ± SD: 233 ± 58 IU/L) on Days 2-7 and 10 (p < 0.001 to p = 0.05, mean ± SD: 307-437 ± 79-146 IU/L). Increased FIB was noted over baseline (mean ± SD: 177 ± 30 mg/dl) on Days 6-8 and 10 (p = 0.02 to p = 0.03, mean ± SD: 234-252 ± 33-49 mg/dl). SAA was increased above baseline (mean ± SD: 4.7 ± 2.9) on Day 6 (p = 0.02, mean ± SD: 113 ± 186 µg/ml). There was no change in HAP. MAIN LIMITATIONS: Healthy horses were used, small sample size, and a lack of a negative control group. CONCLUSIONS: Serial intramuscular procaine penicillin G (IM PPG) injections may result in increased positive APP concentrations in horses and this must be considered when these test results are interpreted.

Evaluation of a point-of-care benchtop analyzer for quantitative measurement of C-reactive protein in canine serum and plasma.

BACKGROUND: Canine C-reactive protein (cCRP) is an acute-phase protein that increases dramatically with inflammation and has potential utility in monitoring disease progression and response to treatment. Rapid, automated point-of-care test (POCT) formats could enhance the clinical utility of cCRP measurement. OBJECTIVES: We aimed to evaluate the VetChroma canine-specific POCT assay for the quantitative measurement of cCRP in canine serum or plasma. METHODS: Serum and plasma from discarded canine diagnostic samples were used. Evaluation included intra- and inter-assay coefficients of variation and observed total error (TEobs ), linearity and spike recovery, the effect of interfering substances and sample matrices, and a method comparison study. RESULTS: Intra-assay variation ranged from 2.5%-6.1%, and inter-assay variation ranged from 2.1%-5.4%. The TEobs ranged from 15.1%-19.7%. The assay was linear over the manufacturer's analytical range with no evidence of constant or proportional bias. Recovery of purified cCRP from canine serum ranged from 116.2% to 138.4%. Hemolysis, icterus, and turbidity did not interfere with the assay. The comparison of paired plasma and serum samples revealed constant and proportional bias. Comparison of the VetChroma cCRP assay to a commercial cCRP ELISA revealed significantly different results. CONCLUSIONS: The VetChroma cCRP assay has acceptable test performance to measure serum cCRP concentration. The POCT protocol and test kit are valid for clinical use, although results obtained using other cCRP assays or plasma may not be directly compared.

Comparison of selected serum biochemistry measurements between the Nova Prime Plus VET, Nova pHOx Ultra, and Beckman Coulter AU680 analyzers in dogs.

BACKGROUND: Blood gas chemistry analyzers typically produce results faster and use smaller sample volumes than reference chemistry analyzers. However, results may not be comparable between blood gas chemistry analyzers and reference chemistry analyzers or between different models of blood gas chemistry analyzers. This could suggest the use of separate reference intervals and, thus, has implications when making clinical decisions. OBJECTIVE: We aimed to perform method comparison studies to evaluate selected canine serum biochemical values obtained using the Nova Stat Profile Prime Plus VET (Prime Plus VET), Stat Profile Nova pHOx Ultra (Ultra), and Beckman Coulter AU680 (Beckman) analyzers. We hypothesized that the three analyzers would be identical within inherent imprecision. METHODS: Jugular venous blood samples were collected from 103 endurance-trained sled dogs, and serum was harvested and stored for analysis. Results for serum chloride, potassium, sodium, creatinine, and urea nitrogen concentrations obtained from the Prime Plus VET and Ultra analyzers were compared with results from the Beckman analyzer, which was considered to be a reference method. Results for serum chloride, potassium, sodium, creatinine, urea nitrogen, and L-lactate concentrations obtained from the Prime Plus VET and Ultra analyzers were compared. Passing-Bablok regression and Bland-Altman plots were used for method comparison. RESULTS: Significant (P < 0.05) constant or proportional bias was found for many analytes for all three method comparison studies. CONCLUSIONS: Due to the presence of statistically significant differences between all three analyzers that may be clinically relevant, it is recommended that reference intervals be created for new blood gas analyzers, even when similar methodologies are used.

The effect of age and sex on selected hematologic and serum biochemical analytes in 4,804 elite endurance-trained sled dogs participating in the Iditarod Trail Sled Dog Race pre-race examination program.

Endurance-trained sled dogs provide a unique translational model to characterize changes in hematologic and serum biochemical analytes due to the aging process. The primary objective of this study was to determine the effect of age and sex on specific hematologic and serum biochemical parameters in the endurance trained sled dog. Longitudinal and cross-sectional data were analyzed from 9,746 blood and serum samples from 4,804 dogs collected over 7 years as part of the Iditarod Trail Sled Dog Race pre-race examination program. Mixed models analysis was used for statistical analysis and P < 0.01 was considered significant. Dogs ranged from 1-12 years of age and 39% were female. Serum total calcium and phosphorus concentrations and white blood cell count decreased nonlinearly to asymptotic values by 6.6, 3.1, and 6.9 years of age, respectively, equivalent to estimated physiologic ages in human years of 44, 27, and 46 years. Serum glucose concentrations reached their lowest value at 7.8 years of age, equivalent to an estimated human physiologic age of 50 years, after which time the concentration increased. Serum globulin concentrations increased with age, but nonlinearly for females and linearly for males. Most sex-related differences were <5%; however, females had lower serum urea nitrogen (14.7%) and creatinine (7.3%) concentrations, lower serum alanine aminotransferase activity (16.6%), and higher serum total bilirubin concentration (12.8%) and platelet count (6.0%). The endurance-trained sled dog provides an excellent model to separate the physiologic effects of age from those of a sedentary lifestyle on hematologic and serum biochemical analytes.

Mapping career patterns in research: A sequence analysis of career histories of ERC applicants.

Despite the need to map research careers, the empirical evidence on career patterns of researchers is limited. We also do not know whether career patterns of researchers can be considered conventional in terms of steady progress or international mobility, nor do we know if career patterns differ between men and women in research as is commonly assumed. We use sequence analysis to identify career patterns of researchers across positions and institutions, based on full career histories of applicants to the European Research Council frontier research grant schemes. We distinguish five career patterns for early and established men and women researchers. With multinomial logit analyses, we estimate the relative likelihood of researchers with certain characteristics in each pattern. We find grantees among all patterns, and limited evidence of gender differences. Our findings on career patterns in research inform further studies and policy making on career development, research funding, and gender equality.

The utility of intraoperative impression smear cytology of intracranial granular cell tumors: Three cases.

Two adult male dogs (a 7-year-old shorthaired Chihuahua and 14-year-old Shih Tzu) and one adult female dog (a 9-year-old Maltese) presented for evaluation of new-onset seizure activity. Magnetic resonance imaging of the brain demonstrated a large, poorly marginated T2-weighted hyperintense, and strong contrast enhancing extra-axial mass in each case. A surgical biopsy for histopathologic evaluation was elected in all cases, and intraoperative impression smears were successfully obtained. Intraoperative cytology identified a homogenous population of round to polygonal cells with central to eccentric nuclei, coarse chromatin, and variably amphophilic to eosinophilic granular cytoplasm. Cytologic findings led to a suspected diagnosis of granular cell tumor (GCT) in all cases. Histopathologic review identified a densely cellular, unencapsulated neoplastic mass comprised of sheets of large round to polygonal cells with abundant eosinophilic cytoplasm containing numerous eosinophilic intracytoplasmic granules, confirming the diagnosis of GCT in all cases. The cases reported here are unique in that they reveal an accurate intraoperative cytologic diagnosis of a rare canine central nervous system neoplasm. Intraoperative cytology of the intracranial masses could provide clinicians with important and quick diagnostic and prognostic information; therefore, expediting decisions made intraoperatively. Further research is warranted to determine the diagnostic accuracy of intraoperative cytology for neoplasia in veterinary patients.

Basal cell enumeration does not predict malignancy in canine perianal gland tumor cytology.

BACKGROUND: Perianal (hepatoid) gland tumors are common in dogs, and the distinction between the benign and malignant forms is clinically important. Cytology of these tumors typically contains many large hepatoid cells and fewer small basal cells. OBJECTIVES: The objective of this study was to determine whether the proportion of the smaller basaloid reserve cells in cytologic samples from perianal tumors correlates with malignancy. METHODS: Eighty-three cases of cytologically diagnosed perianal gland tumors with corresponding histopathologic sections were identified from two separate institutions and included six (7.2%) malignant tumors and 77 (92.8%) benign tumors. The proportion of basal cells from each sample was evaluated. RESULTS: No difference between these groups was found, although the study was sufficiently powered to detect an approximately 1.5-fold change in basal cell proportion. CONCLUSIONS: This report found no evidence that the proportion of basal cells in canine perianal tumor cytology is an indication of the potential for malignancy. We, therefore, do not recommend citing this feature in cytologic reports or when communicating with clinicians.

Integrated and isolated impact of high-performance work practices on employee health and well-being: A comparative study.

We investigate the positive relationships between high-performance work practices (HPWP) and employee health and well-being and examine the conflicting assumption that high work intensification arising from HPWP might offset these positive relationships. We present new insights on whether the combined use (or integrated effects) of HPWP has greater explanatory power on employee health, well-being, and work intensification compared to their isolated or independent effects. We use data from the 2004 British Workplace Employment Relations Survey (22,451 employees nested within 1,733 workplaces) and the 2010 British National Health Service Staff survey (164,916 employees nested within 386 workplaces). The results show that HPWP have positive combined effects in both contexts, and work intensification has a mediating role in some of the linkages investigated. The results also indicate that the combined use of HPWP may be sensitive to particular organizational settings, and may operate in some sectors but not in others. (PsycINFO Database Record

"Managing" the Placebo Effect: The Single-Blind Placebo Lead-in Response in Two Pain Models.

OBJECTIVE: "Placebo effects" in analgesic medication trials for chronic pain are pervasive; however, little is known regarding mechanisms or factors that may influence the presence or magnitude of these effects. Our objective is to consider elements of the placebo response in the context of two pain models using a "single-blind placebo lead-in" design (SBPLI). METHODS: As part of two pilot drug trials using knee osteoarthritis (KOA) and non-radicular low back pain (LBP) subjects, SBPLI protocols were conducted. We examined whether gender and/or diagnosis affected placebo responses as observed in changes in patient self-reported pain, depressive and pain anxiety symptoms. We also evaluated the placebo response on performance-based tests (stair climbing, range of motion (ROM), sit to stand repetitions, and 6-minute treadmill distance). RESULTS: VAS Pain Intensity (Now) values decreased significantly during the SBPLI for the sample as a whole, but the effects appeared stronger among LBP subjects. CES-D short form values (depressive symptoms) did not decrease significantly during the SBPLI for the sample as a whole, but some placebo effects appeared to emerge for women in the KOA group. PASS values (pain anxiety symptoms) decreased significantly, albeit mildly, during the SBPLI for the sample as a whole.Stair Climb (time) revealed no significant SBPLI effects. Bend Forward to Floor (ROM) increased significantly at the end of the SBPLI period for the sample as a whole, but the effects appeared stronger among KOA subjects. Sit to Stand Repetitions increased during the SBPLI for the sample as a whole. Treadmill Distance did not change significantly from Visit 1 to Visit 2; however, a significant Sex difference for the KOA group was found such that women showed greater Treadmill Distance at Visit 2. CONCLUSION: Placebo effects emerged across psychometric and performance-based measures, indicating the pervasiveness of this phenomenon. In this design, diagnostic and (to a lesser extent) gender categories differentials were observed during the placebo period. The SBPLI design may prove not only a robust method in studying the placebo phenomena, but also as a design element to mitigate some aspects of the placebo response in clinical trials.

Cerebellar associative sensory learning defects in five mouse autism models.

Sensory integration difficulties have been reported in autism, but their underlying brain-circuit mechanisms are underexplored. Using five autism-related mouse models, Shank3+/ΔC, Mecp2(R308/Y), Cntnap2-/-, L7-Tsc1 (L7/Pcp2(Cre)::Tsc1(flox/+)), and patDp(15q11-13)/+, we report specific perturbations in delay eyeblink conditioning, a form of associative sensory learning requiring cerebellar plasticity. By distinguishing perturbations in the probability and characteristics of learned responses, we found that probability was reduced in Cntnap2-/-, patDp(15q11-13)/+, and L7/Pcp2(Cre)::Tsc1(flox/+), which are associated with Purkinje-cell/deep-nuclear gene expression, along with Shank3+/ΔC. Amplitudes were smaller in L7/Pcp2(Cre)::Tsc1(flox/+) as well as Shank3+/ΔC and Mecp2(R308/Y), which are associated with granule cell pathway expression. Shank3+/ΔC and Mecp2(R308/Y) also showed aberrant response timing and reduced Purkinje-cell dendritic spine density. Overall, our observations are potentially accounted for by defects in instructed learning in the olivocerebellar loop and response representation in the granule cell pathway. Our findings indicate that defects in associative temporal binding of sensory events are widespread in autism mouse models.

A systematic review of ketamine for complex regional pain syndrome.

OBJECTIVE: This systematic review aims to examine the available literature and to synthesize published data concerning the treatment of Complex Regional Pain Syndrome (CRPS) with ketamine. METHODS: The search was conducted utilizing the databases Medline, Embase and the Cochrane Central Registry of Controlled Trials. All relevant articles were systematically reviewed. RESULTS: The search yielded 262 articles, 45 of which met the inclusion/exclusion criteria. Of those included, 6 were reviews, 5 were randomized placebo-controlled trials, 13 were observational studies, and 21 were case reports. CONCLUSION: There is no high quality evidence available evaluating the efficacy of ketamine for CRPS and all manuscripts examined in this review were of moderate to low quality. Therefore, we conclude there is currently only weak evidence supporting the efficacy of ketamine for CRPS, yet there is clearly a rationale for definitive study.

Biochemical survey of free-roaming cats (Felis catus) in New York City presented to a trap-neuter-return program.

Free-roaming cats in New York, NY, USA, that presented to a trap-neuter-return program were surveyed for biochemical data. One hundred and one cats had blood collected for a plasma biochemistry panel after the induction of surgical anesthesia. Reference intervals for 18 analytes were generated for the sample population, along with age-specific reference intervals when statistically appropriate. Age groups (juveniles and adults) differed in 10 of the 18 analytes measured, including protein levels and albumin/globulin ratio, aspartate aminotransferase, alkaline phosphatase, creatine kinase, creatinine, phosphorus, calcium and potassium. No differences were found between males and females. This is the first report of biochemical reference intervals for a group of free-roaming cats within the USA.

Sodium nitroprusside is not associated with metabolic acidosis during intraoperative infusion in children.

BACKGROUND: Sodium nitroprusside (SNP) is a potent vasodilator that has been used to induce deliberate hypotension in children during surgery involving significant blood loss, including craniofacial and spinal fusion procedures. SNP metabolism liberates cyanide, which may cause interference with cellular energy metabolism, leading to metabolic acidosis and central nervous system injury. We performed a retrospective, case-control study to determine whether the short-term intra-operative use of SNP for deliberate hypotension is associated with metabolic acidosis in children undergoing surgical procedures for craniofacial or spinal anomalies. Cyanide and thiocyanate concentrations were also recorded in patients who received SNP. METHODS: Data from 166 children undergoing craniofacial and spinal fusion surgery between 2005 and 2010 at Lucile Packard Children's Hospital (LPCH) at Stanford were analyzed. Records from 60 patients who received SNP (SNP group) as part of a multicenter, randomized, double-blind study were compared with records from 106 eligible patients who had blood pressure reduction using anesthetic agents and did not receive SNP (control group). Metabolic acidosis was defined as serum bicarbonate (HCO3) < 18.5 mEq/L. Whole blood CN, plasma thiocyanate and urinary thiocyanate concentrations were measured in patients in the SNP group. Differences in metabolic acidosis rates between the SNP and control groups were assessed through a test of noninferiority in the rate for the SNP group with a noninferiority threshold of 0.2. A z-test was used to test the null hypothesis. The alternative hypothesis was that the difference in these rates was less than 0.2. The same noninferiority threshold of 0.2 was also used to perform separate, secondary tests for noninferiority in the proportion of patients with HCO3 levels below 18.5 mEq/L and the proportion of patients who required HCO3 administration. RESULTS: Fewer patients in the SNP group experienced metabolic acidosis compared to the control group (31.7% vs. 36.8%, respectively; p < .001). No whole blood CN levels above the lower limit of quantification were detected in any of the 51 patients with validated CN data. Plasma and urinary thiocyanate levels were also low. CONCLUSIONS: Our findings suggest that SNP, when used for short-term deliberate hypotension, does not cause an increased incidence of metabolic acidosis compared with the use of anesthetic agents alone. TRIAL REGISTRATION NUMBER: NCT00135668.

Dual infection with Pythium insidiosum and Blastomyces dermatitidis in a dog.

A 4-year-old male neutered Labrador Retriever with severe gastrointestinal signs, but no respiratory signs, was diagnosed with multifocal pyogranulomatous gastritis, enteritis, and lymphadenitis with intralesional hyphae and multifocal pyogranulomatous pneumonia with intralesional yeast. Based on cytologic evaluation, histologic examination with special stains, and immunohistochemical analysis of tissues collected antemortem or at necropsy, dual infections with Pythium insidiosum and Blastomyces dermatitidis were detected and are reported for the first time.

Speed kills? Not always: the New York State thruway experience.

BACKGROUND: The New York State (NYS) speed limit increased from 55 mph to 65 mph in August 1995. This study examines the change in motor vehicle fatality rates on the NYS thruway and NYS Interstates before and after the speed limit change. We also compare the fatality rates pre- and postincreased speed limit between the NYS thruway and other major NYS roadways, where speed limit remained unchanged at 55 mph. METHODS: The number of fatalities and vehicle miles traveled (VMT) on individual roadways were extracted from NYS Department of Transportation reports. The data were divided into groups by prespeed limit change, August 1992 to August 1995, and postspeed limit change, August 1995 to August 1998, and roads where the speed limit remained unchanged at 55 mph. Data were also analyzed to assess the effect of variability of speed and general topography on changes in fatality rates on roads where the speed limit was increased. RESULTS: There was a 28.3% decrease in absolute mortality and a 42.6% decrease in mortality adjusted for VMT, on the NYS thruway after the speed limit increase from 55 mph to 65 mph. When compared with roadways, where the speed limit remained unchanged, there was a 52.4% reduction in fatality rates below the predicted rates after the speed limit increase (p = 0.009). On the NYS interstate system, there was a 13.5% decrease in absolute mortality and a 29.2% decrease in mortality adjusted for VMT, after the speed limit increase. The percentage of traffic traveling >10 mph faster than the speed limit dropped from 39% to 8% on roads, where the speed limit was increased from 55 mph to 65 mph. In the NYS interstate system, "flat" roads (elevation <200 m) had a 30.2% reduction in absolute mortality, whereas "mountainous" roads (elevations >200 m) had a 17.6% increase in absolute mortality in response to the speed limit increase (p < 0.001). CONCLUSION: Motor vehicle speed is a key determinate of the severity of injury in an individual crash; however, speed variance and road topography have also been found to impact the total number of motor vehicle fatalities in the NY State highway system.