The effects of dietary essential fatty acid ratios and linoleic acid level in grow-finish pigs.

The objective of this study was to investigate the effects of dietary linoleic acid level and the ratio of linoleic acid:linolenic acid (LA:ALA) on the growth performance, expression of genes associated with lipid metabolism, and inflammatory status of grow-finish pigs. A total of 300 growing pigs (body weight [BW] = 41.1 ±â€…6.3 kg) were randomly assigned to either a high (30 g/kg; HLA) or low (15 g/kg; LLA) dietary linoleic acid level with a high (23:1; HR), moderate (13:1; MR) or low (4:1; LR) dietary LA:ALA in a 2 × 3 factorial design. Diets were fed across three 28-d phases and were balanced for dietary metabolizable energy. Pigs were housed five pigs per pen in single-sex pens. Blood samples were collected on days 0, 21, 42, and 84, and synovial fluid was collected from the hock joint on days 0 and 84 for inflammatory marker analysis. Data were analyzed as repeated measures using PROC MIXED (SAS 9.4) with initial BW as a covariate, pen as the experimental unit, and LA level, LA:ALA, sex, phases, and their interactions as fixed effects. Compared to HLA, LLA pigs tended to have increased BW at days 56 and 84 (P = 0.088). There was no effect of LA × LA:ALA for growth performance. For the overall days 0 to 84 growth period, pigs fed HR had increased ADG compared to MR, with pigs receiving LR performing intermediate of MR and HR. Gilts receiving HR diets had increased day 84 BW compared to gilts receiving the low and moderate LA:ALA (P = 0.006), which was a result of improved overall days 0 to 84 ADG compared to gilts receiving the MR diets (P = 0.023). Barrows fed LR had improved BW on day 56 compared to MR and HR and higher final BW compared to HR, with MR performing intermediately (P = 0.006). This was a result of greater days 0 to 84 ADG (P = 0.023). Overall, C-reactive protein (CRP), tumor necrosis factor-α (TNFα), and interleukin-6 were reduced in the plasma of pigs over time (P ≤ 0.037). Across all treatments, CRP and TNFα were reduced in the hock and carpus synovial fluid on day 84 vs. day 0 (P ≤ 0.049). In conclusion, LA:ALA ratios utilized in this study can be fed at varying linoleic acid levels without impacting growth or inflammation. Additionally, LA:ALA ratios can differentially impact the growth of gilts and barrows.

Previous research in lactating sows has reported that dietary inclusion of the essential fatty acids linoleic acid and linolenic acid is important for performance. Research in grow­finish pigs has shown an improvement in gilt growth performance when fed differing linoleic:linolenic acid ratios (LA:ALA); however, further research evaluating LA:ALA in diets with similar metabolizable energy is needed in growing pigs. In the present research, a 23:1 dietary essential fatty acid ratio increased the final body weight of gilts compared to a 13:1 or 4:1 LA:ALA, while barrows fed a 4:1 dietary essential fatty acid ratio had increased gain and final body weight compared to a 23:1 LA:ALA. Plasma and synovial fluid inflammatory markers were also reduced with time and were unaffected by dietary LA:ALA or linoleic acid inclusion. Dietary essential fatty acid ratio can differentially impact the growth of barrows and gilts, with no impact on systemic or joint inflammation.

The effects of dietary essential fatty acid ratios and energy level on growth performance, lipid metabolism, and inflammation in grow-finish pigs.

The objective of this study was to investigate the effects of dietary metabolizable energy (ME) level and the ratio of linoleic acid:α-linolenic acid (LA:ALA) on the growth performance, lipid metabolism, circulatory and joint inflammatory status, and synovial fluid proteome of grow-finish pigs. A total of 224 pigs (BW = 41.5 ±â€…6.1 kg; PIC Genus 337 × 1050, Hendersonville, TN) were randomly assigned to either a high (3.55 Mcal/kg; HE) or low (3.29 Mcal/kg; LE) ME dietary treatment with a high (23:1) or low (12:1) LA:ALA in a 2 × 2 factorial arrangement. Diets were fed across three 28-d phases. Pigs were housed either four barrows or four gilts per pen. Blood samples were collected on days 0, 21, 42, and 84. Synovial fluid was collected from the hock and carpus joints on days 0 and 84. Liver and adipose tissue samples were collected on day 84. Data were analyzed as repeated measures using PROC MIXED (SAS 9.4) with pen as the experimental unit and energy level, essential fatty acid ratio, sex, phase, and their interactions as fixed effects. Compared to LE, HE increased days 28, 56, and 84 body weight (BW; P = 0.005). For the overall period, HE increased average daily gain (ADG) compared to LE (P < 0.001) and improved feed efficiency (P = 0.001), while LE increased feed intake compared to HE (P < 0.001). Gilts receiving diets with low LA:ALA had similar final BW to barrows receiving a low LA:ALA at days 28, 56, and 84 (P = 0.024), resulting from improved overall days 0-84 ADG compared to gilts receiving the high LA:ALA (P = 0.031). In the liver, HE decreased the mRNA abundance of acetyl CoA carboxylase (ACACA; P = 0.004), cluster of differentiation 36 (P = 0.034), and tended to decrease fatty acid synthase (FASN; P = 0.056). In adipose tissue, HE decreased ACACA (P = 0.001) and FASN (P = 0.017). Plasma inflammatory markers C-reactive protein (CRP) and tumor necrosis factor-α (TNFα) were reduced on day 84 compared to day 0 (P ≤ 0.014). In the hock and carpus synovial fluid, LE tended to reduce CRP and TNFα (P ≤ 0.096). Hock and carpus synovial fluid CRP were also reduced on day 84 compared to day 0 (P = 0.001). Age of the pig impacted serum and hock synovial fluid protein abundance, but not energy level, LA:ALA, or their interactions (P < 0.05). To conclude, the high and low LA:ALA ratios utilized in this study can be fed at varying energy levels without impacting growth. Additionally, LA:ALA ratios can differentially impact the growth of barrows and gilts.

In pig diets, it has been established that added fat can improve growth and feed efficiency; however, insufficient research has been reported evaluating specific essential fatty acids found in commonly available fat sources. Essential fatty acids are important in several biological functions in the body, including growth, inflammation, and immune function. Given shared metabolism between essential fatty acids linoleic acid and α-linolenic acid, it has been suggested that their dietary ratio is critical to balance inflammatory responses. In the present research, a 12:1 dietary linoleic:linolenic acid ratio improved gilt, but not barrow, daily gain and did not impact inflammation. Pro-inflammatory responses were reduced over time, both in the blood and joint fluid. High-fat diets also improved growth performance, suppressed genes involved in fatty acid synthesis, and tended to increase joint inflammation. There was no interaction between dietary fat level and essential fatty acid ratio for any variable. Overall, dietary essential fatty acid ratios impact the growth of gilts, regardless of dietary fat inclusion, with no apparent effects on inflammation.

Characterization of in-barn heat processed swine mortalities.

In-barn heat processing of mass swine mortalities to inactivate pathogens could facilitate more carcass disposal options and reduce the risk of pathogen spread in the event of a foreign animal disease (FAD) outbreak. A 12.2 × 12.2 × 2.4 m (W × L × H) heat processing room was created using a temporary wall inside a de-commissioned commercial gestation barn in northwest Iowa. Eighteen swine carcasses (six per group) divided into three weight groups (mean ± SD initial carcass weights: 31.8 ± 3.3, 102.7 ± 8.1, and 226.3 ± 27.6 kg) were randomly assigned a location inside the room. Three carcasses per weight group were placed directly on concrete slats and on a raised platform. One carcass per weight group and placement (n=6) was instrumented with five temperature sensors, inserted into the brain, pleura, peritoneal, ham, and bone marrow of the femur, and a sensor was attached directly to the skin surface. Environmental conditions (ambient and room) and carcass temperatures were collected at 15-min intervals. Carcasses were subjected to an average room temperature of 57.3 ± 1.2°C for 14 days. The average (±SD) reduction from initial weight for the carcasses on slats was 45.0 ± 4.70% (feeder), 33.0 ± 8.30% (market), and 34.0 ± 15.80% (sow), and for the carcasses on a raised platform, it was 39.0 ± 6.80% (feeder), 49.0 ± 11.30% (market), and 45.0 ± 6.70% (sow). There was a significant interaction between carcass placement (slats and raised) and carcass weight loss for the market weight group. When average carcass surface temperature was at 40.6, 43.3, and 46.1°C (data grouped for analysis), the average internal carcass temperature for most measurement locations was significantly different across carcass weight groups and between the carcasses on a raised platform and those on slats. This preliminary analysis of carcass weight loss, leachate production, and temperature variation in carcasses of different sizes can be used for planning and evaluating mass swine mortality management strategies.

Barriers and Facilitators to Pre-exposure Prophylaxis Uptake Among Black Women: A Qualitative Analysis Guided by a Socioecological Model.

ABSTRACT: Black women experience disparities in HIV incidence. Pre-exposure prophylaxis (PrEP) is a once-daily pill that can prevent HIV transmission. To enhance PrEP uptake among Black women, it is essential to examine their perceptions of PrEP. In 2018, 33 Black women in New York City completed interviews about their attitudes, knowledge, and perceived barriers and facilitators to PrEP use. Emergent themes were organized using a socioecological model. Participants identified barriers at the sociocultural level, including stigma, medical mistrust, and health care avoidance. At the community level, health care access issues and limited community knowledge were reported. Partner-level barriers included trust in partners and meaning attributed to PrEP use within the context of monogamy. Individual-level barriers included low perceived risk and concerns about PrEP's safety and efficacy. Our findings can inform future PrEP research with Black women, as well as PrEP implementation efforts aimed at increasing uptake among this population.

Development and Improvement of an International Webcast Series to Expand the Accessibility of Swine Medicine Resources.

Swine medicine resources and caseloads for teaching and supporting extracurricular training activities vary widely among veterinary colleges and are concentrated in specific regions. Student interest and demand for swine medicine training is broader in geographical distribution. This is illustrated by student membership and attendance at the American Association of Swine Veterinarians (AASV) annual meetings, for example. To explore how concentrated resources might be made more widely available in a cost-effective manner, the Swine Medicine Education Center (SMEC) at Iowa State University's College of Veterinary Medicine looked for ways to leverage existing extracurricular resources with a broader geography of schools and students. This article describes the organization of student chapters of the AASV and the outcomes of a multi-session live audio and video webcast focused on swine medicine topics across North America over a 3-year period. SMEC organized the series with funding provided by the AASV and AASV Foundation. The broadcast series covered a wide range of swine-related topics, including pet pigs, emerging diseases, and regulation of antimicrobials. In its third year, 25 North American and 4 international veterinary schools participated in the series and provided feedback from attendees.

Pharmacokinetics of Intravenous, Intramuscular, Oral, and Transdermal Administration of Flunixin Meglumine in Pre-wean Piglets.

Castration and tail-docking of pre-wean piglets are common procedures that are known to induce pain and would benefit from pain mitigation. Flunixin meglumine (FM) is a non-steroidal anti-inflammatory drug currently approved in the United States for pyrexia in swine and lameness pain in cattle. The objective of this study was to establish the pharmacokinetic (PK) parameters resulting from intravenous (IV), intramuscular (IM), oral (PO) and transdermal (TD) administration of FM in pre-wean piglets. FM was administered to thirty-nine pre-wean piglets at a target dose of 2.2 mg/kg for IV and IM and 3.3 mg/kg for PO and TD route. Plasma was collected at twenty-seven time points from 0 to 9 days after FM administration and concentrations were determined using ultra-high performance liquid chromatography coupled with mass spectrometry (UPLC-MS). Pharmacokinetic data were analyzed using noncompartmental analysis (NCA) methods and nonlinear mixed-effects (NLME). Initial plasma concentration for IV (C0) 11,653 µg/L and mean peak plasma concentrations (Cmax) 6,543 µg/L (IM), 4,883 µg/L (PO), and 31.5 µg/L (TD) were measured. The time points of peak FM concentrations (tmax) were estimated 30 min, 1 h, and 24 h for IM, PO, and TD, respectively. The bioavailability (F) of PO and IM FM was estimated at >99%, while the bioavailability of TD FM was estimated to be 7.8%. The reported Cmax of FM after IM and PO administration is consistent with therapeutic concentration ranges that mitigate pain in other species and adult pigs. However, the low estimated concentration of FM after TD dosing is not expected to mitigate pain in pre-wean piglets. The low F of TD FM suggests that expanding the surface area of application is unlikely to be sufficient to establish an effective TD dose for pain, while the high bioavailability for PO FM should allow for an effective dose regimen to be established.

Analytical Validation of a Highly Sensitive, Multiplexed Chronic Myeloid Leukemia Monitoring System Targeting BCR-ABL1 RNA.

This study describes the analytical performance of the QuantideX qPCR BCR-ABL IS Kit, the first Food and Drug Administration-cleared assay designed to monitor breakpoint cluster region-Abelson tyrosine-protein kinase 1 (BCR-ABL1) fusion transcripts isolated from peripheral blood specimens from patients with chronic myeloid leukemia. This multiplex real-time quantitative RT-PCR assay amplifies both e13a2 and e14a2 Major BCR-ABL1 transcripts and the reference target ABL1. The test results are provided in international scale (IS) values by incorporating armored RNA-based calibrators that have defined IS values tied directly to the World Health Organization BCR-ABL1 Primary Reference Materials, without the necessity of determining and maintaining conversion factors. For each batch run, the integrated interpretive software evaluates run and specimen quality control metrics (including a sufficient amount of ABL1 control transcripts to ensure a minimal limit of detection) and calculates both molecular response (MR) and %IS values for each specimen. The test has a limit of detection of MR4.7 (0.002%IS) and a linear range from MR0.3 (50%IS) to MR4.7 (0.002%IS) for both Major transcripts. Single-site and multisite precision studies demonstrated a maximum SD of 0.13 MR (30% CV within the assay range between MR0.7 and MR3.7). The performance of this BCR-ABL1 monitoring test meets all of the clinical guideline recommendations for sensitivity and IS reporting for the management of chronic myeloid leukemia patients.

The effect of chemical clarification of oral fluids on porcine epidemic diarrhea virus antibody responses.

Routine testing of breeding herd oral fluid (OF) samples for porcine epidemic diarrhea virus (PEDV) IgG and/or IgA is used to track levels of PEDV immunity over time. However, OFs contain particles of feed, feces, and inorganic material that detract from the quality of the sample. We clarified swine OF samples using lyophilized chitosan-based formulas (A-C) tested by PEDV IgG and IgA ELISAs. To evaluate both the immediate and residual effects of treatment on antibody detection, samples were tested immediately post-treatment, then stored at 4°C and retested at 2, 4, and 6 days post-treatment (DPT). Formulations were shown to effectively clarify samples. Statistical analysis comparing treated to untreated OF samples at 0 DPT found that neither chitosan nor Tween 20 affected the OF ELISA IgA and IgG sample-to-positive (S/P) ratio results ( p > 0.05). Furthermore, pairwise comparisons of 0 DPT to 2, 4, and 6 DPT results detected no significant differences ( p > 0.05) in IgA and IgG S/P ratios (i.e., treated OF samples were stable over time). Therefore, chitosan efficiently clarified OF specimens without affecting the results of the PEDV IgG and IgA antibody ELISAs.

Validation of a next-generation sequencing assay for clinical molecular oncology.

Currently, oncology testing includes molecular studies and cytogenetic analysis to detect genetic aberrations of clinical significance. Next-generation sequencing (NGS) allows rapid analysis of multiple genes for clinically actionable somatic variants. The WUCaMP assay uses targeted capture for NGS analysis of 25 cancer-associated genes to detect mutations at actionable loci. We present clinical validation of the assay and a detailed framework for design and validation of similar clinical assays. Deep sequencing of 78 tumor specimens (≥ 1000× average unique coverage across the capture region) achieved high sensitivity for detecting somatic variants at low allele fraction (AF). Validation revealed sensitivities and specificities of 100% for detection of single-nucleotide variants (SNVs) within coding regions, compared with SNP array sequence data (95% CI = 83.4-100.0 for sensitivity and 94.2-100.0 for specificity) or whole-genome sequencing (95% CI = 89.1-100.0 for sensitivity and 99.9-100.0 for specificity) of HapMap samples. Sensitivity for detecting variants at an observed 10% AF was 100% (95% CI = 93.2-100.0) in HapMap mixes. Analysis of 15 masked specimens harboring clinically reported variants yielded concordant calls for 13/13 variants at AF of ≥ 15%. The WUCaMP assay is a robust and sensitive method to detect somatic variants of clinical significance in molecular oncology laboratories, with reduced time and cost of genetic analysis allowing for strategic patient management.

Rapid, semi-automated, and inexpensive radioimmunoassay of cAMP: application in GPCR-mediated adenylate cyclase assays.

Cyclic AMP (cAMP) is an important signal transduction second messenger that is commonly used as a functional mirror on the actions of G protein-coupled receptors that can activate or inhibit adenylate cyclases. A radioimmunoassay for cAMP with femtomole sensitivity was first reported by Steiner more than 30 years ago, and there have been several subsequent modifications that have improved this assay in various ways. Here we describe additional improvement to existing methods that markedly improve speed and reduce cost without sacrificing sensitivity, and is also adaptable to analysis of cGMP. The primary antibody is coupled directly to magnetic beads that are then separated from unbound marker using filtration on microplates. This eliminates the need for a secondary antibody, and markedly increases throughput. In addition, we report a simple, reproducible, and inexpensive method to make the radiomarker used for this assay. Although still requiring the use of radioactivity, the resulting method retains a high degree of accuracy and precision, and is suitable for low-cost high throughput screening. Use of aspects of this method can also improve throughput in other radioimmunoassays.

Functional selectivity of dopamine D1 receptor agonists in regulating the fate of internalized receptors.

Recently, we demonstrated that D(1) agonists can cause functionally selective effects when the endpoints of receptor internalization and adenylate cyclase activation are compared. The present study was designed to probe the phenomenon of functional selectivity at the D(1) receptor further by testing the hypothesis that structurally dissimilar agonists with efficacies at these endpoints that equal or exceed those of dopamine would differ in ability to influence receptor fate after internalization, a functional endpoint largely unexplored for the D(1) receptor. We selected two novel agonists of therapeutic interest that meet these criteria (the isochroman A-77636, and the isoquinoline dinapsoline), and compared the fates of the D(1) receptor after internalization in response to these two compounds with that of dopamine. We found that dopamine caused the receptor to be rapidly recycled to the cell surface within 1h of removal. Conversely, A-77636 caused the receptor to be retained intracellularly up to 48 h after agonist removal. Most surprisingly, the D(1) receptor recovered to the cell surface 48 h after removal of dinapsoline. Taken together, these data indicate that these agonists target the D(1) receptor to different intracellular trafficking pathways, demonstrating that the phenomenon of functional selectivity at the D(1) receptor is operative for cellular events that are temporally downstream of immediate receptor activation. We hypothesize that these differential effects result from interactions of the synthetic ligands with aspects of the D(1) receptor that are distal from the ligand binding domain.

Polymorphisms in the glucocerebrosidase gene and pseudogene urge caution in clinical analysis of Gaucher disease allele c.1448T>C (L444P).

BACKGROUND: Gaucher disease is a potentially severe lysosomal storage disorder caused by mutations in the human glucocerebrosidase gene (GBA). We have developed a multiplexed genetic assay for eight diseases prevalent in the Ashkenazi population: Tay-Sachs, Gaucher type I, Niemann-Pick types A and B, mucolipidosis type IV, familial dysautonomia, Canavan, Bloom syndrome, and Fanconi anemia type C. This assay includes an allelic determination for GBA allele c.1448T>C (L444P). The goal of this study was to clinically evaluate this assay. METHODS: Biotinylated, multiplex PCR products were directly hybridized to capture probes immobilized on fluorescently addressed microspheres. After incubation with streptavidin-conjugated fluorophore, the reactions were analyzed by Luminex IS100. Clinical evaluations were conducted using de-identified patient DNA samples. RESULTS: We evaluated a multiplexed suspension array assay that includes wild-type and mutant genetic determinations for Gaucher disease allele c.1448T>C. Two percent of samples reported to be wild-type by conventional methods were observed to be c.1448T>C heterozygous using our assay. Sequence analysis suggested that this phenomenon was due to co-amplification of the functional gene and a paralogous pseudogene (PsiGBA) due to a polymorphism in the primer-binding site of the latter. Primers for the amplification of this allele were then repositioned to span an upstream deletion in the pseudogene, yielding a much longer amplicon. Although it is widely reported that long amplicons negatively impact amplification or detection efficiency in recently adopted multiplex techniques, this assay design functioned properly and resolved the occurrence of false heterozygosity. CONCLUSION: Although previously available sequence information suggested GBA gene/pseudogene discrimination capabilities with a short amplified product, we identified common single-nucleotide polymorphisms in the pseudogene that required amplification of a larger region for effective discrimination.

Stress and coping: the roles of ethnicity and gender in substance abuse recovery.

This study investigated ethnic and gender differences in reported resource losses and gains for recovering substance abusers living in Oxford Houses (OH). Participants (n = 829) completed a version of Hobfoll's (1998) Conservation of Resources (COR) Evaluation. Results indicated significant individual differences in resources, based on gender, ethnicity, and the length of OH residential stay. Men reported fewer resource gains and losses than women. With respect to ethnicity, African-Americans reported greater gains and losses in resources than European-Americans. Individuals with less time in an OH also reported having experienced more losses in the past 3 months.

Community reactions to campaigns addressing crystal methamphetamine use among gay and bisexual men in New York City.

Crystal methamphetamine (aka "crystal meth") use with high-risk sex has become an emerging health problem for gay and bisexual men in New York City since the late 1990s. Public health campaigns were eventually developed to encourage gay and bisexual men to avoid or reconsider using crystal meth. Reactions to three campaigns were measured with a cross-sectional survey administered in 2004. Among an ethnically-diverse sample of 971 gay and bisexual men, 61.8% reported seeing the campaigns. Those who reported ever using crystal meth, recent use, and recent use with sex were significantly more likely to have seen the campaigns. In general, white men, HIV-negative men, and men not currently using crystal meth responded more positively to the campaigns than their counterparts; yet, more men of color reported having discussions with partners and friends about their crystal use as a result of these campaigns. Implications for researchers and practitioners are discussed.

Adoption of array technologies into the clinical laboratory.

Array-based methods are making substantial contributions to the discovery of disease biomarkers and are fueling the growth of multianalyte testing for disease diagnosis and treatment. The distillation of high-density array results into sets of signature markers promises to improve disease staging, risk stratification and treatment decisions. To accommodate the growing requirement for multiplex testing, clinical laboratories are converting several single-analyte tests into array-based formats. However, adoption of array technologies provides several challenges to the laboratory, which must evaluate these new formats, train laboratory personnel, market the new services and obtain reimbursement for new analytes. Liquid-bead arrays are an attractive format for routine clinical diagnostics due to a combination of appropriate analyte density, simultaneous array decoding and detection, and flexibility for rapid customization. In this review, the suitability of several array platforms to diagnostic testing and applications of liquid-bead arrays for cystic fibrosis testing, multidisease carrier status assays and leukemia subtyping are discussed. As our understanding of the clinical utility of new or established biomarkers and recommendations for testing change, flexibility and adaptability of array platforms will be imperative. Future development of novel assay formats and improved quantitation will expand the number of diseases tested and lead to further integration into the diagnostic laboratory.