Arthropods Associated with Invasive Frangula alnus (Rosales: Rhamnaceae): Implications for Invasive Plant and Insect Management.

The invasive shrub glossy buckthorn (Frangula alnus) has been progressively colonizing the Northeastern United States and Southeastern Canada for more than a century. To determine the dominant arthropod orders and species associated with F. alnus, field surveys were conducted for two years across 16 plots within the Allegheny National Forest, Pennsylvania, USA. Statistical analyses were employed to assess the impact of seasonal variation on insect order richness and diversity. The comprehensive arthropod collection yielded 2845 insects and arachnids, with hemipterans comprising the majority (39.8%), followed by dipterans (22.3%) and arachnids (15.5%). Notably, 16.2% of the hemipterans collected were in the immature stages, indicating F. alnus as a host for development. The two dominant insect species of F. alnus were Psylla carpinicola (Hemiptera: Psyllidae) and Drosophila suzukii (Diptera: Drosophilidae); D. suzukii utilized F. alnus fruits for reproduction. Species richness and diversity exhibited significant variations depending on the phenology of F. alnus. The profiles of volatile compounds emitted from the leaves and flowers of F. alnus were analyzed to identify factors that potentially contribute to the attraction of herbivores and pollinators. The results of our study will advance the development of novel F. alnus management strategies leveraging the insects associated with this invasive species.

Predicting dry matter intake in beef cattle.

Technology that facilitates estimations of individual animal dry matter intake (DMI) rates in group-housed settings will improve production and management efficiencies. Estimating DMI in pasture settings or facilities where feed intake cannot be monitored may benefit from predictive algorithms that use other variables as proxies. This study examined the relationships between DMI, animal performance, and environmental variables. Here we determined whether a machine learning approach can predict DMI from measured water intake variables, age, sex, full body weight, and average daily gain (ADG). Two hundred and five animals were studied in a drylot setting (152 bulls for 88 d and 53 steers for 50 d). Collected data included daily DMI, water intake, daily predicted full body weights, and ADG using In-Pen-Weighing Positions and Feed Intake Nodes. After exclusion of 26 bulls of low-frequency breeds and one severe (>3 standard deviations) outlier, the final number of animals used for modeling was 178 (125 bulls, 53 steers). Climate data were recorded at 30-min intervals throughout the study period. Random Forest Regression (RFR) and Repeated Measures Random Forest (RMRF) were used as machine learning approaches to develop a predictive algorithm. Repeated Measures ANOVA (RMANOVA) was used as the traditional approach. Using the RMRF method, an algorithm was constructed that predicts an animal's DMI within 0.75 kg. Evaluation and refining of algorithms used to predict DMI in drylot by adding more representative data will allow for future extrapolation to controlled small plot grazing and, ultimately, more extensive group field settings.

In animal agriculture, passive monitoring technology has the potential to lead to needed innovations as we look for solutions to make global food production more resilient. Here, we use passive intake systems to measure daily weight, water intake, and climatic variables to accurately predict dry matter intake. Such an approach, if it can be successfully applied for grazing animals would dramatically improve the ability of animal agriculture to reduce the ecological footprints of food production. Two hundred and five animals were studied in a drylot setting (152 bulls for 88 d and 53 steers for 50 d). We used both traditional statistical and modern machine learning approaches to test the ability to predict dry matter intake. Although all approaches had success in predicting dry matter intake, the best prediction came from a machine learning approach which was able to predict the average daily dry matter intake during a test to within 0.75 kg/d. Evaluation and refining of algorithms used to predict dry matter intake in the drylot by adding more representative data will allow for future extrapolation to controlled small plot grazing and, ultimately, more extensive grazing animal intakes at a production scale.

Sexual Dimorphism of Skeletal Muscle in a Mouse Model of Breast Cancer: A Functional and Molecular Analysis.

Breast cancer incidence in men is statistically rare; however, given the lack of screening in males, more advanced stages at initial diagnosis result in lower 5-year survival rates for men with breast cancer compared to women. A sexual dimorphism, with respect to the effect of tumor growth on cachexia incidence and severity, has also been reported across cancer types. The purpose of this study was to examine the sexual dimorphism of breast cancer as it pertains to skeletal muscle function and molecular composition. Using female and male transgenic PyMT mice, we tested the hypothesis that the isometric contractile properties and molecular composition of skeletal muscle would be differentially affected by breast tumors. PyMT tumor-bearing mice of each sex, corresponding to maximal tumor burden, were compared to their respective controls. RNA sequencing of skeletal muscle revealed different pathway alterations that were exclusive to each sex. Further, differentially expressed genes and pathways were substantially more abundant in female tumor mice, with only minimal dysregulation in male tumor mice, each compared to their respective controls. These differences in the transcriptome were mirrored in isometric contractile properties, with greater tumor-induced dysfunction in females than male mice, as well as muscle wasting. Collectively, these data support the concept of sexually dimorphic responses to cancer in skeletal muscle and suggest that these responses may be associated with the clinical differences in breast cancer between the sexes. The identified sex-dependent pathways within the muscle of male and female mice provide a framework to evaluate therapeutic strategies targeting tumor-associated skeletal muscle alterations.

Sexual Dimorphism of Skeletal Muscle in a Mouse Model of Breast Cancer: A Functional and Molecular Analysis.

Breast cancer incidence in men is statistically rare; however, given the lack of screening in males, more advanced stages at initial diagnosis results in lower 5-year survival rates for men with breast cancer compared to women. A sexual dimorphism, with respect to the effect of tumor growth on cachexia incidence and severity, has also been reported across cancer types. The purpose of this study was to examine the sexual dimorphism of breast cancer as it pertains to skeletal muscle function and molecular composition. Using female and male transgenic PyMT mice, we tested the hypothesis that isometric contractile properties and molecular composition of skeletal muscle would be differentially affected by breast tumors. PyMT tumor-bearing mice of each sex, corresponding to maximal tumor burden, were compared to their respective controls. RNA-sequencing of skeletal muscle revealed different pathway alterations that were exclusive to each sex. Further, differentially expressed genes and pathways were substantially more abundant in female tumor mice, with only minimal dysregulation in male tumor mice, each compared to their respective controls. These differences in the transcriptome were mirrored in isometric contractile properties, with greater tumor-induced dysfunction in females than male mice, as well as muscle wasting. Collectively, these data support the concept of sexually dimorphic responses to cancer in skeletal muscle and suggest these responses may be associated with the clinical differences in breast cancer between the sexes. The identified sex-dependent pathways within muscle of male and female mice provide a framework to evaluate therapeutic strategies targeting tumor-associated skeletal muscle alterations.

Effects of Forest Management on the Insect Assemblage of Black Cherry (Prunus serotina) in the Allegheny National Forest.

Over the last decade, the Allegheny National Forest (ANF) in the USA has experienced issues with the regeneration of black cherry (Prunus serotina). This study was conducted to investigate the effects of silvicultural treatment on the insect communities that may affect black cherry pollination and regeneration. We conducted a 2-year study to compare the abundance, richness, and diversity of insects in unmanaged, shelterwood seed-tree, and shelterwood clear-cut stands. Using pan traps, we sampled insects at the ground level and in the canopies of flowering mature black cherry trees. The results of this study showed significant increases in the abundance of insects captured in shelterwood seed-tree stands and in species richness and diversity of insects captured in the canopy of black cherry in shelterwood removal stands, indicating that silvicultural treatment affected the insect community significantly. The dominant insect order was Diptera (true flies, 72.91%, n = 12,668), and Anthalia bulbosa (Diptera: Hybotidae) was the dominant species comprising 33% of all insects found in the canopy of flowering black cherry. The findings in this study could help land managers in managing black cherry for its pollination and natural regeneration.

Characterization of the Insect Assemblage and Associated Floral Volatiles of Black Cherry (Prunus serotina).

Black cherry is an ecologically important high-value wood. A decline of its regeneration has been reported in the USA, which could be associated with a lack of pollination. This study was conducted to identify insects visiting black cherry flowers, to determine whether insects captured on the flowers carry black cherry pollen and to identify the volatile organic compounds (VOCs) emitted by flowers of black cherry. A two-year insect survey was conducted before, during and after the black cherry bloom. A total of 9533 insects were captured in traps and Diptera was the most abundant (64.1%). Significantly more insects in Diptera, Lepidoptera and Thysanoptera were captured in the traps installed in the canopy than those on the ground, and Anthalia bulbosa (Diptera: Hybotidae) was the dominant species. Electron microscopy analyses demonstrated that insects captured in the canopy indeed carried black cherry pollen. Black cherry flowers emitted a VOC blend that is composed of 34 compounds and dominated by ß-ocimene and several phenylpropanoids/benzenoids. This floral VOC profile is similar to that of other pollinator-dependent Prunus species. This study reports pollinator insects and associated VOCs, for the first time, that could play a significant role in the pollination and regeneration of black cherry.

Behavioral problems may be associated with multilevel lumbosacral stenosis in military working dogs.

Behavioral and spinal problems have been reported to be important causes of early retirement in military working dogs, however studies on possible relationships between these two problems are currently lacking. The aim of this retrospective, observational, cross-sectional study was to test associations between reported behavioral problems and computed tomographic (CT) diagnoses of lumbosacral (LS) stenosis in a sample of military working dogs. Data archives at a tertiary referral military working dog hospital were searched for dogs that had LS CT scans acquired during the period of June 2013- July 2014, and available medical records. For each included dog, an observer unaware of CT findings reviewed medical records and recorded dog signalment data. A second observer reviewed medical records and assigned a score of 0 (absent) or 1 (present) for each of 11 behavioral problem categories. A third observer unaware of medical record findings reviewed CT scans and assigned a score of 0 (absent) or 1 (present) for LS stenosis at each of 4 vertebral locations (L4-5, L5-6, L6-7, L7-S1). A statistician selected and performed tests of association. A total of 55 dogs met inclusion criteria. Of these, 21 dogs (38.2%) had recorded behavioral complaints/problems involving behavior in at least 1 of the 10 categories and 44 (80.0%) had CT diagnoses of LS stenosis in at least one vertebral location. The number of behavioral problems were significantly associated with number of vertebral locations with LS stenosis (ordinal logistic fit, p=0.011) and positively correlated (rho = 0.37, p = 0.006). Problems involving behavioral concerns were more likely to be exhibited if dogs had multi-level stenosis (≥3 vertebral levels, Kappa = 0.06, p>0.05; Bowker's test statistics 26.26, P<0.05). The most common behavior problems in dogs with multi-level stenosis were "unwilling or reluctant to jump up onto objects/into vehicles" (3/8, 38%), "sudden onset of aggressive behaviors" (2/8, 25%), "self-mutilation in the lower back region, tail, or hind legs "(2/8, 25%), "increase in anxiety" (2/8, 25%), "sudden decrease in appetite" (2/8, 25%), "unwilling or reluctant to sit" (2/8, 25%), and "handler-reported unusual behaviors" (2/8, 25%). Findings from this preliminary study supported inclusion of multi-level LS stenosis in the differential diagnosis list for military working dogs presenting with a recent onset of behavioral problems.

Associations of Adiposity and Diet Quality with Serum Ceramides in Middle-Aged Adults with Cardiovascular Risk Factors.

Rates of adverse cardiovascular events have increased among middle-aged adults. Elevated ceramides have been proposed as a risk factor for cardiovascular events. Diet quality and weight status are inversely associated with several traditional risk factors; however, the relationship to ceramides is less clear. This study aimed to determine associations of adiposity and diet quality with circulating ceramides in middle-aged adults (n = 96). Diet quality was estimated using the Healthy Eating Index 2015 (HEI-2015). Serum ceramide concentrations were determined by liquid chromatography-mass spectrometry. A ceramide risk score was determined based on ceramides C16:0, C18:0, and C24:1 and their ratios to C24:0. Participants who were classified as at 'moderate risk' compared to 'lower-risk' based on a ceramide risk score had significantly higher body mass index (BMI) values, as well as higher rates of elevated fibrinogen levels, metabolic syndrome, and former smoking status. BMI was positively associated with the ceramide C18:0 (R2 = 0.31, p < 0.0001), the ratio between C18:0/C24:0 ceramides (R2 = 0.30, p < 0.0001), and the ceramide risk score (R2 = 0.11, p < 0.009). Total HEI-2015 scores (R2 = 0.42, p = 0.02), higher intakes of vegetables (R2 = 0.44, p = 0.02) and whole grains (R2 = 0.43, p = 0.03), and lower intakes of saturated fats (R2 = 0.43, p = 0.04) and added sugar (R2 = 0.44, p = 0.01) were associated with lower C22:0 values. These findings suggest that circulating ceramides are more strongly related to adiposity than overall diet quality. Studies are needed to determine if improvements in weight status result in lower ceramides and ceramide risk scores.

CT and gross pathology are comparable methods for detecting some degenerative sacroiliac joint lesions in dogs.

Degenerative sacroiliac joint disease is a cause of lumbosacral pain in dogs; however, published information on cross-sectional imaging characteristics is limited. Objectives of this retrospective, secondary analysis, methods-comparison study were to test hypotheses that CT lesions reported in humans with degenerative sacroiliac joint disease are also present in dogs, and that CT is comparable to gross pathology for detecting these lesions. Matched CT and gross pathology slice images of 30 sacroiliac joints were retrieved from a previous prospective, canine cadaver study. A veterinary radiologist interpreted randomized CT images for each joint based on previously published CT characteristics of lesions in humans with degenerative sacroiliac joint disease. A veterinary pathologist independently interpreted randomized gross pathology images using the same criteria. All joints had at least one CT lesion consistent with degenerative sacroiliac joint disease. A new CT lesion was also identified and termed "subarticular cleft." The CT and gross pathology methods agreed for detecting joints with subchondral sclerosis, subchondral erosion, and intra-articular ankylosis lesions (P > .05, McNemar's test), but disagreed for detection of joints with subchondral cyst, para-articular ankylosis, and subarticular cleft lesions (P ≤ .05). Using gross pathology as the reference standard, CT had 100% sensitivity for detection of subarticular cleft and subchondral cyst lesions, with 56% and 22% specificity, respectively. Para-articular ankylosis lesions were detected by CT but not by gross pathology. Findings supported the hypothesis that CT lesions reported in humans with degenerative sacroiliac joint disease are also present in dogs, and partially supported the hypothesis that CT is comparable to gross pathology for detecting joints with these lesions.

Nutrition Risk is Associated with Leukocyte Telomere Length in Middle-Aged Men and Women with at Least One Risk Factor for Cardiovascular Disease.

Poor diet quality has been associated with several age-related chronic conditions, but its relationship to telomere length, a biological marker of cellular aging, is unclear. The purpose of this cross-sectional study was to determine whether overall diet quality was associated with relative leukocyte telomere length (rLTL) in a sample (n = 96) of nonsmoking middle-aged adults in Appalachia with at least one risk factor for cardiovascular disease. Diet quality was assessed using the Healthy Eating Index (HEI-2015), the alternate Mediterranean diet score (aMed), and the Dietary Screening Tool (DST). Peripheral rLTL was measured by quantitative real-time polymerase chain reaction. The associations between potentially confounding sociodemographic, lifestyle and health-related factors and the first and fourth rLTL quartile groups were examined using Chi-square or Fisher's Exact tests or logistic regression. The relationships between diet quality index scores and rLTL as a continuous variable were analyzed using simple linear regression and multivariate linear models, analogous to linear covariance analyses. The rLTL ranged from 0.46 to 1.49 (mean ± SEM was 1.02 ± 0.18). Smoking history, income level, and cardiovascular health (Life's Simple 7) were associated with the lowest and highest quartiles of rLTL and were used as covariates. In adjusted and unadjusted models, participants considered "at nutrition risk" by the DST were more likely to have shorter rLTL than those "not at risk or at potential risk" (p = 0.004). However, there was no evidence that adherence to the 2015⁻2020 Dietary Guidelines for Americans or to a Mediterranean diet was associated with rLTL in this sample. Intervention studies are needed to determine if improving the diet quality of those at nutrition risk results in reduced telomere attrition over time.

Educational intervention improves fruit and vegetable intake in young adults with metabolic syndrome components.

The FRUVEDomics study investigates the effect of a diet intervention focused on increasing fruit and vegetable intake on the gut microbiome and cardiovascular health of young adults with/at risk for metabolic syndrome (MetS). It was hypothesized that the recommended diet would result in metabolic and gut microbiome changes. The 9-week dietary intervention adhered to the US Department of Agriculture Dietary Guidelines for Americans and focused on increasing fruit and vegetable intake to equal half of the diet. Seventeen eligible young adults with/or at high risk of MetS consented and completed preintervention and postintervention measurements, including anthropometric, body composition, cardiovascular, complete blood lipid panel, and collection of stool sample for microbial analysis. Participants attended weekly consultations to assess food logs, food receipts, and adherence to the diet. Following intention-to-treat guidelines, all 17 individuals were included in the dietary, clinical, and anthropometric analysis. Fruit and vegetable intake increased from 1.6 to 3.4 cups of fruits and vegetables (P < .001) daily. Total fiber (P = .02) and insoluble fiber (P < .0001) also increased. Clinical laboratory changes included an increase in sodium (P = .0006) and low-density lipoprotein cholesterol (P = .04). In the fecal microbiome, Erysipelotrichaceae (phylum Firmicutes) decreased (log2 fold change: -1.78, P = .01) and Caulobacteraceae (phylum Proteobacteria) increased (log2 fold change = 1.07, P = .01). Implementing a free-living 9-week diet, with intensive education and accountability, gave young adults at high risk for/or diagnosed with MetS the knowledge, skills, and feedback to improve diet. To yield greater impact, a longer diet intervention may be needed in this population.

Long-term Immunotoxic Effects of Oral Prenatal and Neonatal Atrazine Exposure.

Atrazine and its metabolites are present at high concentrations in many water supplies in agro-intensive areas. Because residents in these areas drink water from sources fed from these contaminated supplies, we investigated the long-term immunotoxicity of combined prenatal and neonatal (perinatal) exposure to atrazine via drinking water, on the immune system in mice. At 6 months of age, upon immunization with heat-killed Streptococcus pneumoniae, the serum IgG antibody response against the T independent antigen phosphorylcholine was significantly higher in male, but not female, atrazine-exposed mice as compared with that in untreated controls. No alterations were present in all offspring in the serum antibody response against the T-dependent antigen pneumococcal surface protein A (PspA). ELISpot analysis showed only a small, insignificant reduction in PspA-specific IgG producing splenocytes in atrazine-treated male offspring. Interestingly, upon ex vivo stimulation with anti-CD3 and anti-CD28 antibodies, significant decreases in interleukin (IL)-2, tumor necrosis factor-α, interferon-γ, and IL-17A and a decreasing trend in IL-10 were observed in splenocytes from atrazine-exposed male, but not female mice. Analysis of thymic and splenic cell populations showed no effects of atrazine exposure in either sex. This is the first time that long-term changes in the immune response were observed after a perinatal exposure to atrazine and it demonstrates that these early life exposures can result in permanent changes to the immune system as well as a male bias in these effects.

Breed-specific differences in the immune response to lipopolysaccharide in ewes.

Innate immune response to a lipopolysaccharide (LPS) challenge varies among sheep breeds. How different breeds respond to bacterial infections impacts management practices of sheep producers. Hence, clinical response, acute-phase response, and gene expression of pro- and anti-inflammatory markers in peripheral white blood cells (WBCs) were examined after an LPS challenge in Dorset and Suffolk ewes. Ewes received either PBS or 2.5 µg/kg LPS (i.v.) 4 to 5 d after onset of synchronized estrus. Blood was collected via jugular venipuncture intermittently for 24 h to determine WBC counts. Rectal temperatures and observations of behavioral/physical appearances were recorded hourly. After LPS, WBCs decreased the first hour (P = 0.0001) and rectal temperatures (P < 0.0001) increased through 4 h; both returned toward normal 6 h after challenge. Suffolk ewes exhibited greater changes in temperature (P = 0.03) and behavioral/physical responses (P < 0.0001) than Dorset ewes and had an enhanced acute-phase response demonstrated by increased concentrations of plasma haptoglobin (P = 0.04), as well as cortisol concentrations (P = 0.03). Real-time PCR was completed on buffy coat homogenates for expression of pro-inflammatory [CXCL8, IL-6, interferon gamma (IFNG), complement component 3 (C3), toll-like receptor 4 (TLR4), prostaglandin synthase 2 (PTGS2)] and anti-inflammatory [IL-10, superoxide dismutase 2 (SOD2), forkhead box P3 (FOXP3), peroxisome proliferator-activated receptor gamma (PPARG), mannose receptor C type 1 (MRC1), transforming growth factor ß (TGFß)] genes. After LPS treatment, gene expressions increased for CXCL8 (P = 0.0003), TLR4 (P = 0.004), SOD2 (P < 0.0001), and C3 (P = 0.003), while PPARG (P = 0.006) and MRC1 (P = 0.003) decreased. Overall, Dorset ewes had greater expression of TLR4 (P = 0.003), IL-10 (P = 0.045), PPARG (P = 0.002), FOXP3 (P = 0.001), and SOD2 (P = 0.0002), whereas Suffolk ewes had greater expression of IL-6 (P = 0.0007), IFNG (P = 0.02), PTGS2 (P = 0.0002), and C3 (P = 0.008). Suffolk ewes also displayed greater expression of IL-6 (P = 0.002) and C3 (P = 0.0004) in response to LPS. In conclusion, differences in gene expression may explain the enhanced inflammatory response in Suffolk ewes and may predispose Suffolk ewes to be more responsive to bacterial infection than Dorset ewes.

Neck Circumference Positively Relates to Cardiovascular Risk Factors in College Students.

The objective of this study was to determine the relationship between neck circumference (NC) and other anthropometric measures and examine cut-off points for males and females according to existing waist circumference cut-off levels in this age group. Across 8 universities, 1562 students underwent a physical assessment. Spearman rho correlations (ρ) were calculated to determine associations between NC and other continuous variables of health. Receiving operating characteristic curves were constructed to assess the optimal cut-off levels of NC of males and females with central obesity. Participants were predominantly Caucasian (67%), female (70%), and outside of Appalachia (82%). Forty-one percent of males and 34% of females had a BMI ≥ 25 kg/m². In both sexes, significant positive correlations were seen between NC and body weight, BMI, waist circumference, hip circumference, and systolic blood pressure (all p-values < 0.0001). NC ≥ 38 cm for males and ≥33.5 cm for females were the optimal cut-off values to determine subjects with central obesity. NC has been identified to closely correlate with other anthropometric measurements related to disease and could be used as a convenient, low-cost, and noninvasive measurement in large-scale studies.

Shortening of the Pittsburgh Sleep Quality Index Survey Using Factor Analysis.

OBJECTIVE/INTRODUCTION: Lengthy surveys have the potential to burden users and can lead to inaccuracies. Conducting analyses to shorten existing validated surveys is beneficial. The objective, therefore, was to shorten the Pittsburgh Quality Sleep Index (PSQI) for young adults. METHODS: PSQI data from 1246 college students were used. An exploratory factor analysis (FA) was utilized to shorten survey after dropping select items. Nonparametric correlation analysis (Spearman's rho) was conducted between the global sleep scores of the shortened and original surveys. Agreements tests (Kappa and McNemar's test) measured the agreement of the surveys and sensitivity and specificity were evaluated. RESULTS: Six factors were examined using maximum likelihood factoring method, applying squared multiple correlations with Promax rotation to allow for correlated variables. FA with six factors explained 100% of shared variance based on eigenvalues and accounted for 61% of variability based on variables. The FA resulted in 13 selected questions ("shortPSQI"), corresponding to 5 of the 7 components of the original survey. High correlation was found between the global scores of the original survey and the "shortPSQI" (rho = 0.94, p < 0.001). When the global score was converted to the categorical variable of good or poor sleepers, the agreement test indicated strong agreement (Kappa 0.83, 95% CI 0.79-0.86, p < 0.0001). CONCLUSION: The validated, 19-item PSQI survey was shortened to 13 items. Tests of correlation and agreement indicate the "shortPSQI" may be an acceptable alternative to the original survey for young adults. CLINICAL TRIAL REGISTRATION: Data for this study was taken from the Get Fruved study, registered on October 21, 2016, on clinicaltrials.gov (NCT02941497).

Phenotyping of lumbosacral stenosis in Labrador retrievers using computed tomography.

Deep phenotyping tools for characterizing preclinical morphological conditions are important for supporting genetic research studies. Objectives of this retrospective, cross-sectional, methods comparison study were to describe and compare qualitative and quantitative deep phenotypic characteristics of lumbosacral stenosis in Labrador retrievers using computed tomography (CT). Lumbosacral CT scans and medical records were retrieved from data archives at three veterinary hospitals. Using previously published qualitative CT diagnostic criteria, a board-certified veterinary radiologist assigned dogs as either lumbosacral stenosis positive or lumbosacral stenosis negative at six vertebral locations. A second observer independently measured vertebral canal area, vertebral fat area, and vertebral body area; and calculated ratios of vertebral canal area/vertebral body area and vertebral fat area/vertebral body area (fat area ratio) at all six locations. Twenty-five dogs were sampled (lumbosacral stenosis negative, 11 dogs; lumbosacral stenosis positive, 14 dogs). Of the six locations, cranial L6 was the most affected by lumbosacral stenosis (33%). Five of six dogs (83%) with clinical signs of lumbosacral pain were lumbosacral stenosis positive at two or more levels. All four quantitative variables were significantly smaller at the cranial aspects of the L6 and L7 vertebral foramina than at the caudal aspects (P < 0.0001). Fat area ratio was a significant predictor of lumbosacral stenosis positive status at all six locations with cranial L6 having the greatest predictive value (R2 = 0.43) and range of predictive probability (25-90%). Findings from the current study supported the use of CT as a deep phenotyping tool for future research studies of lumbosacral stenosis in Labrador retrievers.

Feasibility for Measuring Transverse Area Ratios and Asymmetry of Lumbosacral Region Paraspinal Muscles in Working Dogs Using Computed Tomography.

OBJECTIVES: Describe computed tomographic (CT) anatomy of canine lumbosacral (LS) paraspinal muscles, a method for measuring paraspinal muscle transverse area ratios and asymmetry using CT, and application of this method in a small sample of working dogs with versus without LS pain. METHODS: Published anatomy references and atlases were reviewed and discrepancies were resolved by examination of anatomic specimens and multiplanar reformatted images to describe transverse CT anatomy of LS region paraspinal muscles. Sixteen Belgian malinois military working dogs were retrospectively recruited and assigned to LS pain positive versus negative groups based on medical record entries. A single observer unaware of dog group measured CT transverse areas of paraspinal muscles and adjacent vertebral bodies, in triplicate, for L5-S1 vertebral locations. A statistician compared muscle transverse area ratios and asymmetry at each vertebral location between groups. RESULTS: The relative coefficient of variation for triplicate CT area measurements averaged 2.15% (N = 16). Multifidus lumborum (L6-7), psoas/iliopsoas (L5-6, L6-7), and sacrocaudalis dorsalis lateralis (L6-7, L7-S1) transverse area ratios were significantly smaller in dogs with LS pain (n = 11) versus without LS pain (n = 5) (p ≤ 0.05). Muscle asymmetry values were not significantly greater in dogs with versus without LS pain. CLINICAL RELEVANCE: Computed tomographic morphometry of LS region paraspinal muscles is a feasible objective method for use in future evidence-based research studies in working dogs. Potential future research applications include determining whether decreased paraspinal muscle area ratios and/or increased paraspinal muscle asymmetry could be used as markers for preclinical LS pain in stoic dogs or risk factors for other injuries in high performance canine athletes, or determining whether core muscle strengthening exercise prescriptions for dogs with LS pain have an effect on paraspinal muscle area ratios and asymmetry.

Suppression of arthritis-induced bone erosion by a CRAC channel antagonist.

OBJECTIVE: We have shown in vitro and in vivo that osteoclast maturation requires calcium-release activated calcium (CRAC) channels. In inflammatory arthritis, osteoclasts mediate severe and debilitating bone erosion. In the current study, we assess the value of CRAC channels as a therapeutic target to suppress bone erosion in acute inflammatory arthritis. METHODS: Collagen-induced arthritis (CIA) was induced in mice. The CRAC channel inhibitor 3,4-dichloropropionaniline (DCPA) and a placebo was administered 1 day prior to collagen II booster to induce arthritis. Effects on swelling, inflammatory cell invasion in joints, serum cytokines and bone erosion were measured. RESULTS: Assays, by blinded observers, of arthritis severity showed that DCPA, 21 mg/kg/day, suppressed arthritis development over 3 weeks. Bone and cartilage damage in sections of animal feet was reduced approximately 50%; overall swelling of joints was reduced by a similar amount. Effects on bone density by µCT showed clear separation in DCPA-treated CIA animals from CIA without treatment, while differences between controls without CIA and CIA treated with DCPA differed by small amounts and in most cases were not statistically different. Response was not related to anticollagen titres. There were no adverse effects in the treated group on animal weight or activity, consistent with low toxicity. The effect was maximal 12-17 days after collagen booster, during the rapid appearance of arthritis in untreated CIA. At 20 days after treatment (day 40), differences in arthritis score were reduced and tumour necrosis factor α, interleukin (IL)-1, or IL-6 in the serum of the animals were similar in treated and untreated animals. CONCLUSIONS: DCPA, a novel inhibitor of CRAC channels, suppresses bone erosion associated with acute arthritis in mice and might represent a new treatment modality for acute arthrits.

The XX Sex Chromosome Complement is Required in Male and Female Mice for Enhancement of Immunity Induced by Exposure to 3,4-Dichloropropionanilide.

PROBLEM: The chemical propanil enhances antibody responses to a heat-killed Streptococcus pneumoniae (HKSP) vaccine. The enhanced response is dependent on gonads in females, but independent of gonads in males. The sex differences in the immune response may be due to sexual differentiation of the immune system or sex chromosome complement. METHOD OF STUDY: To test the hypothesis that the immune system is sexually differentiated, newborn C57BL/6 pups were treated with testosterone propionate (TP) or placebo. The role of sex chromosome complement was investigated using the 4-core genotypes (FCG) model of XXF and XYF gonadal females (ovaries), and XXM and XYM gonadal males (testes). For some experiments, mice were gonadectomized or sham gonadectomized. All mice were vaccinated with HKSP, treated with propanil, and the antibody response determined at day seven. RESULTS: Neonatal TP did not alter the response to HKSP. In FCG mice, propanil significantly enhanced the immune response in XXF females and XXM males, but not in XYF females or XYM males. CONCLUSION: The immune system of females was not masculinized by neonatal TP treatment. Sex chromosome complement significantly contributes to the sexually dimorphic immune response after propanil exposure.

The toxicity of the N-hydroxy and 6-hydroxy metabolites of 3,4-dichloropropionanilide does not depend on calcium release-activated calcium channel inhibition.

Each year ~1 billion kg of herbicides are used worldwide to control the unwanted growth of plants. In the United States, over a quarter of a billion kg of herbicides are used, representing 28% of worldwide use. (Kiely, T., Donaldson, D., and Grube, A. [2004]. Pesticide Industry Sales and Usage. 2000 and 2001 Market Estimates. Available at: http://www.epa.gov/pesticides/pestsales/01pestsales/market_estimates2001.pdf. Accessed October 25, 2012.) Propanil (3,4-dichloropropionanilide [DCPA]) is a commonly used herbicide in the United States, with 2-4 million kg applied annually to 2 million acres of crop land. The immunomodulatory effects of DCPA have been well documented, but limited data are available on the effects of its metabolites. (Salazar, K. D., Ustyugova, I. V., Brundage, K. M., Barnett, J. B., and Schafer, R. [2008]. A review of the immunotoxicity of the pesticide 3,4-dichloropropionanalide. J. Toxicol. Environ. Health B Crit. Rev. 11, 630-645.) In mammals, hepatic enzymes metabolize DCPA, resulting in the production of 3,4-dichloroaniline (DCA). Further biotransformation of DCA leads to the production of 6-hydroxy-3,4-dichloroaniline (6OH-DCA) and N-hydroxy-3,4-dichloroaniline (NOH-DCA). We report, for the first time, the immunotoxic effects of DCPA metabolites on T-cell function. Human Jurkat T cells were exposed to varying concentrations of DCPA or its metabolites and assayed for effects on T-cell function. In addition, fluorine analogs of DCPA and DCA were investigated to determine the relative role of chlorine substituents on T-cell immunotoxicity. Here we report that exposure of Jurkat T cells to DCPA and DCA alters IL-2 secretion, nuclear factor of activated T cells (NFAT) activity, and calcium influx. However, exposure to 6OH-DCA and NOH-DCA reduces IL-2 secretion and NFAT activity but has no effect on calcium flux. When both chlorines in DCPA and DCA were substituted with fluorines all effects were abrogated. Our data indicate that metabolites of DCPA have differential effects on T-cell function and the presence of chlorines plays an important role in eliciting these effects.