Robenacoxib shows efficacy for the treatment of chronic degenerative joint disease-associated pain in cats: a randomized and blinded pilot clinical trial.

The main objective of this pilot clinical trial was to evaluate outcome measures for the assessment of the nonsteroidal anti-inflammatory drug (NSAID) robenacoxib in cats with degenerative joint disease-associated pain (DJD-pain). Otherwise healthy cats (n = 109) with DJD-pain entered a parallel group, randomized, blinded clinical trial. Cats received placebo (P) or robenacoxib (R) for two consecutive 3-week periods. Treatment groups were PP, RR, and RP. Actimetry and owner-assessment data were collected. Data were analyzed using mixed-effects and generalized mixed-effects linear models. Activity data showed high within-cat and between-cat variability, and 82.4% of the values were zero. Compared to placebo, mean total activity was higher (5.7%) in robenacoxib-treated cats (p = 0.24); for the 80th percentile of activity, more robenacoxib-treated cats had a > 10% increase in activity after 3 (p = 0.046) and 6 weeks (p = 0.026). Robenacoxib treatment significantly decreased owner-assessed disability, (p = 0.01; 49% reduction in disability; effect size ~ 0.3), and improved temperament (p = 0.0039) and happiness (p = 0.021) after 6 weeks. More robenacoxib-treated cats were successes at 6 weeks (p = 0.018; NNT: 3.8). Adverse effect frequencies were similar across groups. Results identified suitable endpoints for confirmatory studies, while also indicating efficacy of robenacoxib in cats with DJD-pain.

Dog importation and changes in canine intestinal nematode prevalence in Colorado, USA, 2013-2017.

BACKGROUND: Animal rescue and animal welfare organizations are relocating thousands of dogs per year following natural disasters and in attempts to provide greater adoption opportunities. Many dogs are sourced from the southeastern USA, which historically has a high prevalence rate for many parasites and parasitic diseases. The Colorado Department of Agriculture Pet Animal Care Facilities Act (PACFA) requires animal shelters and animal welfare organizations to report annually a variety of statistics including the numbers of dogs imported into Colorado from out of state. The Companion Animal Parasite Council (CAPC) provides data nationally, down to the state and county level, on a variety of common parasitic and vector borne diseases. These data make it possible to track changes in parasite prevalence over several years. METHODS: Test results for canine roundworm, hookworm and whipworm were collected from the CAPC maps for 2013-2017. Dog importation data for 2014-2017 was collected from PACFA reports. For evaluation of the statistical significance of prevalence changes when comparing 2013 to 2017, 2 × 2 contingency tables were constructed with both positive and negative test results for each year and the data assessed using Chi-square tests to determine if the 2017 prevalence was significantly different than the 2013 prevalence for roundworm, hookworm and whipworm. RESULTS: Significant increases in intestinal nematode prevalence occurred in Colorado from 2013 to 2017. The prevalence of canine roundworm rose 35.60%, the prevalence of canine hookworm rose 137.33% and the prevalence of canine whipworm rose 63.68%. From 2014 to 2017, over 114,000 dogs were transported into Colorado from out-of-state, by more than 130 animal shelters and rescue organizations. Three of the larger organizations reported that the majority of their dogs were obtained from New Mexico, Texas and Oklahoma. Texas and Oklahoma have historically much higher parasite prevalence than Colorado. CONCLUSIONS: Veterinarians in areas with historically low parasite prevalence where dogs from high parasite prevalence areas are arriving for adoption may need to reevaluate their recommendations regarding fecal examination and deworming frequencies as historic levels of intestinal parasite infection may no longer be accurate assessments of future infection risks.

Novel use of an activity monitor to model jumping behaviors in cats.

OBJECTIVE: To develop methods to identify and characterize activity monitor (AM) data signatures for jumps performed by cats. ANIMALS: 13 healthy, client-owned cats without evidence of osteoarthritis or degenerative joint disease. PROCEDURES: Each cat was fitted with the same AM, individually placed in an observation room, then simultaneously recorded by 3 video cameras during the observation period (5 to 8 hours). Each cat was encouraged to jump up (JU), jump down (JD), and jump across (JA) during the observation period. Output from the AM was manually annotated for jumping events, each of which was characterized by functional data analysis yielding relevant coefficients. The coefficients were then used in linear discriminant analysis to differentiate recorded jumps as JUs, JDs, or JAs. To assess the model's ability to distinguish among the 3 jump types, a leave-one-out cross-validation method was used, and the misclassification error rate of the overall categorization of the model was calculated. RESULTS: Of 731 jumping events, 29 were misclassified. Overall, the mean misclassification error rate per cat was 5.4% (range, 0% to 12.5%), conversely indicating a correct classification rate per cat of 94.6%. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the model was successful in correctly identifying JUs, JDs, and JAs in healthy cats. With advancements in AM technology and data processing, there is potential for the model to be applied in clinical settings as a means to obtain objective outcome measures.

Dog importation and changes in heartworm prevalence in Colorado 2013-2017.

BACKGROUND: Animal welfare organizations, in order to increase dog adoptions and reduce euthanasia of homeless dogs, are transporting large numbers of dogs from state-to-state. Only approximately 1/3 of animal welfare organizations reportedly test for heartworm infection, treat, or provide heartworm prevention prior to transporting dogs. The reported prevalence of heartworms in dogs in animal shelters or rescued following natural disasters ranges between 14.6-48.8%. This analysis was conducted to evaluate the correlation between dog importation and heartworm prevalence in dogs in Colorado. METHODS: Data were obtained from the Companion Animal Parasite Council (CAPC) interactive heartworm prevalence maps and the Colorado Department of Agriculture Pet Animal Care Facilities Program (PACFA) in order to analyze the relationship between dog importation and the changes in prevalence of heartworm (Dirofilaria immitis) in Colorado from 2013 to 2017. RESULTS: The prevalence of heartworms in Colorado dogs has increased 67.5% between 2013-2017. From 2014 to 2017, more than 114,000 dogs were imported into Colorado by over 130 animal shelters and rescue organizations, which could represent approximately 9.5% of the total estimated 2017 Colorado dog population of nearly 1.2 million dogs. Three large Colorado-based organizations responded to requests for details regarding the originating states from which they received dogs. The majority of these dogs were apparently shipped to Colorado from states with higher heartworm prevalence. New Mexico represented the source of the greatest number of relocated dogs, accounting for just over 30%. Nearly half (49%) of the dogs relocated by these three organizations came from either Texas or Oklahoma. CONCLUSIONS: Animal welfare organizations and veterinarians should increase the testing and prevention of heartworm infections in dogs both prior to, and following, transportation from areas with high heartworm prevalence. Repeated testing is recommended due to the 6-month pre-patent-period associated with D. immitis. Veterinarians and pet owners should increase vigilance with heartworm testing and prevention, even in areas with historically low heartworm prevalence. Movement of dogs from highly endemic areas may increase the risks of local transmission.

Empirical evaluation of consistency and accuracy of methods to detect differentially expressed genes based on microarray data.

BACKGROUND: In this study, we empirically evaluated the consistency and accuracy of five different methods to detect differentially expressed genes (DEGs) based on microarray data. METHODS: Five different methods were compared, including the t-test, significance analysis of microarrays (SAM), the empirical Bayes t-test (eBayes), t-tests relative to a threshold (TREAT), and assumption adequacy averaging (AAA). The percentage of overlapping genes (POG) and the percentage of overlapping genes related (POGR) scores were used to rank the different methods on their ability to maintain a consistent list of DEGs both within the same data set and across two different data sets concerning the same disease. The power of each method was evaluated based on a simulation approach which mimics the multivariate distribution of the original microarray data. RESULTS: For smaller sample sizes (6 or less per group), moderated versions of the t-test (SAM, eBayes, and TREAT) were superior in terms of both power and consistency relative to the t-test and AAA, with TREAT having the highest consistency in each scenario. Differences in consistency were most pronounced for comparisons between two different data sets for the same disease. For larger sample sizes AAA had the highest power for detecting small effect sizes, while TREAT had the lowest. DISCUSSION: For smaller sample sizes moderated versions of the t-test can generally be recommended, while for larger sample sizes selection of a method to detect DEGs may involve a compromise between consistency and power.

Antecedents and mediators of community connection in African American women with breast cancer.

OBJECTIVE: To describe the theory of community connection defined as close relationships with women and men who are members of a neighborhood, a church, a work group, or an organization. Antecedent and mediator variables related to community connection are identified. DESIGN/METHODS: A cross-sectional design was used to assess for relationships among theorized antecedents and mediators of community connection in a sample of 144 African American women aged 21 years and older (mean = 54.9) who had been diagnosed with invasive/infiltrating ductal carcinoma. MEASUREMENT AND ANALYSES: Community connection was measured with the relational health indices-community subscale. Mediator analysis was conducted to assess significance of the indirect effects of the mediator variables, which were fear, breast cancer knowledge, and isolation. RESULTS: Community connection was found to be associated with three of the four antecedents, cancer stigma, stress, and spirituality, but not associated with fatalism. Effects were mediated primarily through fear and isolation with isolation as was more dominant of the two mediators. Surprisingly, breast cancer knowledge showed no significant mediator role. CONCLUSIONS: The importance of isolation and fear as mediators of community connection is highlighted by this research. The study could serve as a model for other researchers seeking to understand connection in ethnic groups and communities.

Significance of margin and extent of dysplasia in loop electrosurgery excision procedure biopsies performed for high-grade squamous intraepithelial lesion in predicting persistent disease.

CONTEXT: High-grade squamous intraepithelial lesions (cervical intraepithelial neoplasia 2 and 3) are commonly treated with loop electrosurgery excision procedure (LEEP) biopsies. OBJECTIVE: To highlight the significance of positive margins and extent of positive margins of the cervical LEEP biopsies in predicting the persistence of high-grade squamous intraepithelial lesion and to provide suggestions for reporting margins in cervical LEEP biopsies. DESIGN: The pathology files at the University of Arkansas for Medical Sciences were searched for cervical intraepithelial neoplasia 2 and 3 treated by LEEP biopsy from 1990 to 2001. RESULTS: A total of 489 LEEP biopsy specimens were retrieved and reviewed; 270 patients had follow-up within 1 year. The biopsy specimens of 110 patients showed positive endocervical margins. One hundred sixty specimens had negative ectocervical-endocervical margins. Follow-up of 54% of the cases with initial positive margins showed residual high-grade squamous intraepithelial lesions. This association was even greater when multiple blocks showed positive endocervical margins and in cases with positive deep margins. On the other hand, a negative margin predicted ability to completely remove the lesion in 95% of patients. CONCLUSION: This study reiterates the significance of the evaluation of the margin, even in samples that were received as multiple fragments. Reporting of LEEP biopsy findings should include the extent of the dysplasia, the status of the ectocervical-endocervical margin, and the status of the deep margin.

Sources of variation in Affymetrix microarray experiments.

BACKGROUND: A typical microarray experiment has many sources of variation which can be attributed to biological and technical causes. Identifying sources of variation and assessing their magnitude, among other factors, are important for optimal experimental design. The objectives of this study were: (1) to estimate relative magnitudes of different sources of variation and (2) to evaluate agreement between biological and technical replicates. RESULTS: We performed a microarray experiment using a total of 24 Affymetrix GeneChip arrays. The study included 4th mammary gland samples from eight 21-day-old Sprague Dawley CD female rats exposed to genistein (soy isoflavone). RNA samples from each rat were split to assess variation arising at labeling and hybridization steps. A general linear model was used to estimate variance components. Pearson correlations were computed to evaluate agreement between technical and biological replicates. CONCLUSION: The greatest source of variation was biological variation, followed by residual error, and finally variation due to labeling when *.cel files were processed with dChip and RMA image processing algorithms. When MAS 5.0 or GCRMA-EB were used, the greatest source of variation was residual error, followed by biology and labeling. Correlations between technical replicates were consistently higher than between biological replicates.

Effect of normalization on significance testing for oligonucleotide microarrays.

MOTIVATION: Normalization techniques are used to reduce variation among gene expression measurements in oligonucleotide microarrays in an effort to improve the quality of the data and the power of significance tests for detecting differential expression. Of several such proposed methods, two that have commonly been employed include median-interquartile range normalization and quantile normalization. The median-IQR method applied directly to fold-changes for paired data also was considered. Two methods for calculating gene expression values include the MAS 5.0 algorithm [Affymetrix. (2002). Statistical Algorithms Description Document. Santa Clara, CA: Affymetrix, Inc. http://www.affymetrix.com/support/technical/whitepapers/sadd-whitepaper.pdf] and the RMA method [Irizarry, R. A., Bolstad, B. M., Collin, F., Cope, L. M., Hobbs, B., Speed, T. P. (2003a). Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res. 31(4,e15); Irizarry, R. A., Hobbs, B., Collin, F., Beazer-Barclay, Y. D., Antonellis, K. J., Scherf, U., Speed, T. P. (2003b). Exploration, normalization, and summaries of high density oligonucleotide array probe-level data. Biostatistics 4(2):249-264; Irizarry, R. A., Gautier, L., Cope, L. (2003c). An R package for analysis of Affymetrix oligonucleotide arrays. In: Parmigiani, R. I. G., Garrett, E. S., Ziegler, S., eds. The Analysis of Gene Expression Data: Methods and Software. Berlin: Springer, pp. 102-119]. RESULTS: In considering these methods applied to a prostate cancer data set derived from paired samples on normal and tumor tissue, it is shown that normalization methods may lead to substantial inflation of the number of genes identified by paired-t significance tests even after adjustment for multiple testing. This is shown to be due primarily to an unintended effect that normalization has on the experimental error variance. The impact appears to be greater in the RMA method compared to the MAS 5.0 algorithm and for quantile normalization compared to median-IQR normalization.

Veterans with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) have a markedly increased rate of second malignancy, which is the most common cause of death.

Patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) have an increased incidence of high-grade lymphoid malignancy. The risk of non-lymphoid second malignancy in this population is not well-defined to date. To test the hypothesis that patients with CLL/SLL have an increased risk of second malignancy, we studied the rate of second malignancy in 132 CLL/SLL patients and compared it to the rate of malignancy (excluding non-melanomatous skin cancer) in the Central Arkansas Veterans Healthcare System population of approximately 38,000 veterans over a period of 11.5 years. The rate of second malignancy, diagnosed concomitantly or after CLL/SLL, and the age-adjusted rate of malignancy calculated from tumor registry reports and demographic data, were used to calculate a Standardized Morbidity Ratio (SMR) with 95% confidence interval (CI). Twenty-one (16%) of the CLL/SLL patients had second malignancies (19 non-lymphoid, 1 Richter's transformation and 1 Hodgkin's disease), which were fatal in 15 (71%) patients. The SMR for the CLL/SLL population was 2.97 (95% CI 1.84-4.55) for second malignancy and 2.69 (95% CI 1.62-4.21) for non-lymphoid second malignancy. This study of a well-defined CLL/SLL population shows a significantly increased risk of second malignancy, which was the primary cause of death for 9% of all CLL/SLL patients (34% of all patient deaths).

Autoimmune cytopenia does not predict poor prognosis in chronic lymphocytic leukemia/small lymphocytic lymphoma.

Chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) is characterized by an acquired immune defect that can cause autoimmune complications, including anemia and thrombocytopenia. We conducted an observational study of the epidemiology, clinical presentation and significance of autoimmune complications of CLL/SLL in 132 patients from a large population (>45,000 veterans), in which at least 90% of patients with CLL/SLL have been previously identified. Over a period of 12.5 years, 12 patients (9.1%) had autoimmune complications; of these, 6 (4.5%) had autoimmune hemolytic anemia (AIHA), 5 (3.8%) had immune thrombocytopenia (ITP), and 1 (0.8%) had pure red blood cell aplasia (PRBA). All 6 cases of AIHA had a positive direct immunoglobulin test for IgG and C3d. In 6 patients, CLL/SLL was an incidental finding at the time of presentation with autoimmune cytopenia. Nine out of 10 patients responded to immunosuppressive therapy, which was complicated by serious infection in 7 cases, one of which was fatal. The major cause of mortality in patients with autoimmune complications of CLL/SLL was secondary malignancy. Survival of patients with immune cytopenia was not significantly different from CLL/SLL patients without immune cytopenia. Among patients with anemia or thrombocytopenia, mortality was significantly higher in those with bone marrow failure compared to an autoimmune etiology. We show that in a non-referred population with a high incidence of CLL/SLL, autoimmune cytopenia can occur early in the natural history of the disease. These data suggest that the Rai and Binet classifications for CLL need to be modified for patients with autoimmune cytopenia. .

Clip migration in stereotactic biopsy.

BACKGROUND: Needle localization breast biopsy (NLBB) is the standard for removal of breast lesions after vacuum assisted core biopsy (VACB). Disadvantages include a miss rate of 0% to 22%, a positive margin rate of approximately 50%, and vasovagal reactions (approximately 20%). We hypothesized that clip migration after VACB is clinically significant and may contribute to the positive margin rates seen after NLBB. METHODS: We performed a retrospective review of postbiopsy films in patients who had undergone VACB with stereotactic clip placement for abnormal mammograms. We measured the distance between the clip and the biopsy site in standard two view mammograms. The location of the biopsy air pocket was confirmed using the prebiopsy calcification site. The Pythagorean Theorem was used to calculate the distance the clip moved within the breast. Pathology reports on NLBB or intraoperative hematoma-directed ultrasound-guided breast biopsy (HUG, which localizes by US the VACB site) were reviewed to assess margin status. RESULTS: In all, 165 postbiopsy mammograms on patients who had VACB with clip placement were reviewed. In 93 evaluable cases, the mean distance the clip moved was 13.5 mm +/- 1.6 mm, SEM (95% CI = 10.3 mm to 16.7 mm). Range of migration was 0 to 78.3 mm. The median was 9.5 mm. In 21.5% of patients the clip was more than 20 mm from the targeted site. Migration of the clip did not change with the age of the patient, the size of the breast or location within the breast. In the subgroup of patients with cancer, margin positivity (including those with close margins) after NLBB was 60% versus 0% in the HUG group. CONCLUSIONS: Significant clip migration after VACB may contribute to the high positive margin status of standard NLBBs. Surgeons cannot rely on needle localization of the clip alone and must be cognizant of potential clip migration. HUG as an alternative biopsy technique after VACB eliminates operator dependency on clip location and may have superior results in margin status.

Neoglycans, carbodiimide-modified glycosaminoglycans: a new class of anticancer agents that inhibit cancer cell proliferation and induce apoptosis.

The soluble form of the syndecan-1 heparan sulfate proteoglycan acts as a tumor suppressor molecule that inhibits growth and induces apoptosis of some cancer cell lines in vitro. Analogs of syndecan-1 were produced by carbodiimide (EDAC) conjugation of glycosaminoglycan (GAG) chains to a protein scaffold, thereby generating synthetic proteoglycans that were evaluated for anticancer properties. Surprisingly, when analyzing activities of the controls, we discovered that EDAC modified GAG chains inhibit myeloma cell viability even in the absence of protein. Here, we describe the production and the activities of these novel molecules called neoglycans. The GAG chains heparin and chondroitin sulfate (CS) were exposed to EDAC to generate the neoglycans neoheparin and neoCS, respectively. Heparin and CS in the absence of EDAC modification have no effect or a slight growth promoting effect on cancer and normal cell lines. However, neoheparin and neoCS substantially reduce cell viability by induction of apoptosis of myeloma and breast cancer cells in vitro. NeoCS when injected directly into breast tumors growing in nude mice reduces or abolishes their growth without causing apparent toxicity to the adjacent normal tissue. The neoglycans need not be continuously present in cell cultures because a short pulse exposure is sufficient to reduce cell viability. NeoCS fractions purified by size exclusion chromatography reduce myeloma cell viability, confirming the specificity of neoglycan activity. Collectively, the results of this study demonstrate the anticancer activities of this new class of GAG chain-based molecules and provide the foundation for future development of neoglycans as novel therapeutic agents.