Publications by authors named "Elizabeth Greene"

95 Publications

Phytogenic Water Additives Improve Broiler Growth Performance via Modulation of Intermediary Metabolism-Related Signaling Pathways.

Animals (Basel) 2021 Mar 9;11(3). Epub 2021 Mar 9.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA.

A ban on the use of antibiotic growth promoters (AGPs) has fueled and promoted scientific research towards the identification of reliable and effective alternatives. The supplementation of phytogenics AV/SSL12 (AVSSL) and Superliv Gold (SG) in water has been shown to improve broiler feed efficiency (FE) via modulation of hypothalamic neuropeptides. However, their effects on peripheral metabolic pathways are still unknown. The present study was undertaken to determine the effects of AVSSL and SG on lipid and protein metabolism-associated pathways in various tissues. Day-old male Cobb 500 chicks ( = 288) were randomly assigned to 3 treatment groups, with 8 replicates of 12 birds each. The treatment groups were fed a basal diet and supplemented with AVSSL or SG in the drinking water at a rate of 2, 4, and 7 mL/100 birds/d during the starter, grower, and finisher phases, respectively. The control group were fed a basal diet with no additive supplementation. On d 35, liver, adipose, and muscle tissue were collected from one bird per pen (8 birds/group). Data were analyzed using Student's T-test to compare one treatment group to the control using Graph Pad Prism version 6.0 for Windows. In the liver, the levels of phosphorylated acetyl-CoA carboxylase alpha (ACCα) were significantly increased in both the AVSSL and SG groups compared to the control. The hepatic expression of sterol regulatory element-binding protein cleavage-activating protein (SCAP) was significantly downregulated in both treated groups compared to the control. AVSSL supplementation downregulated the hepatic expression of SREBP-2 and adiponectin (AdipoQ), while SG administration upregulated hepatic AdipoR1/R2 mRNA abundances compared to the untreated group. Both AVSSL and SG treatments upregulated hepatic stearoyl-CoA desaturase-1 (SCD-1) gene expression compared to their untreated counterparts. In the adipose tissue, the levels of phosphorylated hormone-sensitive lipase (HSL) at Ser855/554 site were increased in both the AVSSL and SG groups compared to the control. However, ATGL protein expression was decreased in SG compared to the untreated group. In the muscle, the levels of phosphorylated mechanistic target of rapamycin (mTOR) were increased in the AVSSL, but decreased in the SG group compared to the control. Collectively, these data indicate that supplementation of the phytogenics AVSSL and SG in water reduced hepatic lipogenesis-related proteins and increased adipose tissue lipolysis- and muscle protein synthesis-associated targets, which might explain, at least partially, the improvement in FE observed in previous research.
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http://dx.doi.org/10.3390/ani11030750DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999555PMC
March 2021

Hypothalamic-pituitary dysfunction in alternating hemiplegia of childhood.

Eur J Paediatr Neurol 2021 Mar 11;32:1-7. Epub 2021 Mar 11.

Division of Pediatric Neurology and Developmental Medicine, Duke Children's Health Center, 2301 Erwin Rd, Durham, NC, 27710, USA. Electronic address:

Background: Many central nervous system disorders result in hypothalamic-pituitary (HP) axis dysfunction. Alternating Hemiplegia of Childhood (AHC) is usually caused by mutations in the ATP1A3 subunit of the Na/K ATPase, predominantly affecting GABAergic interneurons. GABAergic interneurons and the ATP1A3 subunit are both important for function of the hypothalamus. However, whether HP dysfunction occurs in AHC and, if so, how such dysfunction manifests remains to be investigated.

Methods: We conducted a retrospective review of a cohort of 50 consecutive AHC patients for occurrence of HP related manifestations and analyzed the findings of the 6 patients, from that cohort, with such manifestations.

Results: Six out of 50 AHC patients manifested HP dysfunction. Three of these patients were mutation positive and 3 were mutation negative. Of the 6 patients with HP dysfunction, 3 had central precocious puberty. A fourth had short stature due to growth hormone deficiency. Two other patients had recurrent episodes of fever of unknown origin (FUO) diagnosed, after workups, as being secondary to central fever. All patients were evaluated and co-managed by pediatric neurology and endocrinology or rheumatology.

Conclusion: AHC was associated with HP dysfunction in about 12% of patients. Awareness of such dysfunction is important for anticipatory guidance and management particularly in the case of FUO which often presents a diagnostic dilemma. Our findings are also consistent with current understandings of the underlying pathophysiology of AHC and of the HP axis.
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http://dx.doi.org/10.1016/j.ejpn.2021.03.007DOI Listing
March 2021

Hypothalamic-pituitary dysfunction in alternating hemiplegia of childhood.

Eur J Paediatr Neurol 2021 Mar 11;32:1-7. Epub 2021 Mar 11.

Division of Pediatric Neurology and Developmental Medicine, Duke Children's Health Center, 2301 Erwin Rd, Durham, NC, 27710, USA. Electronic address:

Background: Many central nervous system disorders result in hypothalamic-pituitary (HP) axis dysfunction. Alternating Hemiplegia of Childhood (AHC) is usually caused by mutations in the ATP1A3 subunit of the Na/K ATPase, predominantly affecting GABAergic interneurons. GABAergic interneurons and the ATP1A3 subunit are both important for function of the hypothalamus. However, whether HP dysfunction occurs in AHC and, if so, how such dysfunction manifests remains to be investigated.

Methods: We conducted a retrospective review of a cohort of 50 consecutive AHC patients for occurrence of HP related manifestations and analyzed the findings of the 6 patients, from that cohort, with such manifestations.

Results: Six out of 50 AHC patients manifested HP dysfunction. Three of these patients were mutation positive and 3 were mutation negative. Of the 6 patients with HP dysfunction, 3 had central precocious puberty. A fourth had short stature due to growth hormone deficiency. Two other patients had recurrent episodes of fever of unknown origin (FUO) diagnosed, after workups, as being secondary to central fever. All patients were evaluated and co-managed by pediatric neurology and endocrinology or rheumatology.

Conclusion: AHC was associated with HP dysfunction in about 12% of patients. Awareness of such dysfunction is important for anticipatory guidance and management particularly in the case of FUO which often presents a diagnostic dilemma. Our findings are also consistent with current understandings of the underlying pathophysiology of AHC and of the HP axis.
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http://dx.doi.org/10.1016/j.ejpn.2021.03.007DOI Listing
March 2021

Evidence of Mitochondrial Dysfunction in Bacterial Chondronecrosis With Osteomyelitis-Affected Broilers.

Front Vet Sci 2021 1;8:640901. Epub 2021 Feb 1.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR, United States.

A leading cause of lameness in modern broilers is bacterial chondronecrosis with osteomyelitis (BCO). While it is known that the components of BCO are bacterial infection, necrosis, and inflammation, the mechanism behind BCO etiology is not yet fully understood. In numerous species, including chicken, mitochondrial dysfunction has been shown to have a role in the pathogenicity of numerous diseases. The mitochondria is a known target for intracellular bacterial infections, similar to that of common causative agents in BCO, as well as a known regulator of cellular metabolism, stress response, and certain types of cell death. This study aimed to determine the expression profile of genes involved in mitochondrial biogenesis, dynamics, and function. RNA was isolated form the tibias from BCO-affected and healthy broilers and used to measure target gene expression via real-time qPCR. Mitochondrial biogenesis factors PGC-1α and PGC-1β were both significantly upregulated in BCO along with mitochondrial fission factors OMA1, MTFR1, MTFP1, and MFF1 as well as cellular respiration-related genes FOXO3, FOXO4, and av-UCP. Conversely, genes involved in mitochondrial function, ANT, COXIV, and COX5A showed decreased mRNA levels in BCO-affected tibia. This study is the first to provide evidence of potential mitochondrial dysfunction in BCO bone and warrants further mechanistic investigation into how this dysfunction contributes to BCO etiology.
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http://dx.doi.org/10.3389/fvets.2021.640901DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7902039PMC
February 2021

A Phase 2b Study to Evaluate the Safety and Efficacy of VRC01 Broadly Neutralizing Monoclonal Antibody in Reducing Acquisition of HIV-1 Infection in Women in Sub-Saharan Africa: Baseline Findings.

J Acquir Immune Defic Syndr 2021 May;87(1):680-687

Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA.

Background: HIV Vaccine Trials Network 703/HIV Prevention Trials Network 081 is a phase 2b randomized, double-blind, placebo-controlled trial to assess the safety and efficacy of passively infused monoclonal antibody VRC01 in preventing HIV acquisition in heterosexual women between the ages of 18 and 50 years at risk of HIV. Participants were enrolled at 20 sites in Botswana, Kenya, Malawi, Mozambique, South Africa, Tanzania, and Zimbabwe. It is one of the 2 Antibody Mediated Prevention efficacy trials, with HIV Vaccine Trials Network 704/HIV Prevention Trials Network 085, evaluating VRC01 for HIV prevention.

Methods: Intense community engagement was used to optimize participant recruitment and retention. Participants were randomly assigned to receive intravenous VRC01 10 mg/kg, VRC01 30 mg/kg, or placebo in a 1:1:1 ratio. Infusions were given every 8 weeks with a total of 10 infusions and 104 weeks of follow-up after the first infusion.

Results: Between May 2016 and September 2018, 1924 women from sub-Saharan Africa were enrolled. The median age was 26 years (interquartile range: 22-30), and 98.9% were Black. Sexually transmitted infection prevalence at enrollment included chlamydia (16.9%), trichomonas (7.2%), gonorrhea (5.7%), and syphilis (2.2%). External condoms (83.2%) and injectable contraceptives (61.1%) were the methods of contraception most frequently used by participants. In total, through April 3, 2020, 38,490 clinic visits were completed with a retention rate of 96% and 16,807 infusions administered with an adherence rate of 98%.

Conclusions: This proof-of-concept, large-scale monoclonal antibody study demonstrates the feasibility of conducting complex trials involving intravenous infusions in high incidence populations in sub-Saharan Africa.
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http://dx.doi.org/10.1097/QAI.0000000000002649DOI Listing
May 2021

Feasibility and Successful Enrollment in a Proof-of-Concept HIV Prevention Trial of VRC01, a Broadly Neutralizing HIV-1 Monoclonal Antibody.

J Acquir Immune Defic Syndr 2021 May;87(1):671-679

Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA.

Background: The Antibody-Mediated Prevention trials (HVTN 704/HPTN 085 and HVTN 703/HPTN 081) are the first efficacy trials to evaluate whether VRC01, a broadly neutralizing monoclonal antibody targeting the CD4-binding site of the HIV envelope protein, prevents sexual transmission of HIV-1. HVTN 704/HPTN 085 enrolled 2701 cisgender men and transgender (TG) individuals who have sex with men at 26 sites in Brazil, Peru, Switzerland, and the United States.

Methods: Participants were recruited and retained through early, extensive community engagement. Eligible participants were randomized 1:1:1 to 10 mg/kg or 30 mg/kg of VRC01 or saline placebo. Visits occurred monthly, with intravenous (IV) infusions every 8 weeks over 2 years, for a total of 10 infusions. Participants were followed for 104 weeks after first infusion.

Results: The median HVTN 704/HPTN 085 participant age was 28 years; 99% were assigned male sex; 90% identified as cisgender men, 5% as TG women and the remaining as other genders. Thirty-two percent were White, 15% Black, and 57% Hispanic/Latinx. Twenty-eight percent had a sexually transmitted infection at enrollment. More than 23,000 infusions were administered with no serious IV administration complications. Overall, retention and adherence to the study schedule exceeded 90%, and the dropout rate was below 10% annually (7.3 per 100 person-years) through week 80, the last visit for the primary end point.

Conclusions: HVTN 704/HPTN 085 exceeded accrual and retention expectations. With exceptional safety of IV administration and operational feasibility, it paves the way for future large-scale monoclonal antibody trials for HIV prevention and/or treatment.
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http://dx.doi.org/10.1097/QAI.0000000000002639DOI Listing
May 2021

Diffuse reflectance spectroscopy reveals heat stress-induced changes in hemoglobin concentration in chicken breast.

Sci Rep 2021 Feb 11;11(1):3649. Epub 2021 Feb 11.

Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR, 72701, USA.

Heat stress (HS) is devastating to the poultry industry due to its adverse effects on animal well-being and performance. The effects of heat stress are typically measured using a portable i-STAT blood analyzer that quantifies circulatory hemoglobin concentration and other blood chemistry parameters. Here, we used diffuse reflectance spectroscopy (DRS) as a novel non-invasive method to directly determine changes in hematological parameters in the breast tissues of live heat-stressed broilers. Three-week-old male broilers were randomly subjected to two environmental conditions (thermoneutral, TN, 24 °C vs. cyclic heat stress, HS, 35 °C, 12 h/day). Optical spectra were acquired using DRS to monitor breast hemoglobin (Hb) concentration and vascular oxygen saturation (sO) at three time points: at baseline prior to heat stress, 2 days, and 21 days after initiation of HS. While i-STAT did not demonstrate a discernible change due to HS in circulatory hemoglobin, DRS found a significant decrease in breast Hb and sO after exposure to chronic HS. The decrease in sO was found to be due to a decrease in oxygenated hemoglobin concentration, indicating a large increase in oxygen consumption in heat-stressed broilers. Our results demonstrate that DRS could potentially be used to study the effects of HS directly in specific organs of interest, such as the breast and thigh, to improve meat quality.
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http://dx.doi.org/10.1038/s41598-021-83293-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7878772PMC
February 2021

COVID-19 Diagnostic Testing For All - Using Non-Dilutive Saliva Sample Collection, Stabilization and Ambient Transport Devices.

medRxiv 2021 Jan 25. Epub 2021 Jan 25.

COVID-19 testing is not accessible for millions during this pandemic despite our best efforts. Without greatly expanded testing of asymptomatic individuals, contact tracing and subsequent isolation of spreaders remains as a means for control. In an effort to increase RT-PCR assay testing for the presence of the novel beta-coronavirus SARS-CoV-2 as well as improve sample collection safety, GenTegra LLC has introduced two products for saliva collection and viral RNA stabilization: GTR-STM™ (GenTegra Saliva Transport Medium) and GTR-STMdk™ (GenTegra Saliva Transport Medium Direct to PCR). Both products contain a proprietary formulation based on GenTegra's novel "Active Chemical Protection™" (ACP) technology that gives non-dilutive, error-free saliva sample collection using RNA stabilization chemicals already dried in the collection tube. GTR-STM can be used for safer saliva-based sample collection at home (or at a test site). Following saliva collection, the sample-containing GTR-STM can be kept at ambient temperature during shipment to an authorized CLIA lab for analysis. SARS-CoV-2 viral RNA in GTR-STM is stable for over a month at ambient temperature, easily surviving the longest transit times from home to lab. GTR-STM enhances patient comfort, convenience, compliance and reduces infectious virus exposure to essential medical and lab professionals. Alternatively, the GTR-STMdk direct-into-PCR product can be used to improve lab throughput and reduce reagent costs for saliva sample collection and testing at any lab site with access to refrigeration. GTR-STMdk reduces lab process time by 25% and reagent costs by 30% compared to other approaches. Since GTR-STMdk retains SARS-CoV-2 viral RNA stability for three days at ambient temperature, it is optimized for lab test site rather than at home saliva collection. SARS-COV-2 viral RNA levels as low as 0.4 genome equivalents/uL are detected in saliva samples using GTR-STMdk. The increased sensitivity of SARS-CoV-2 detection can expand COVID-19 testing to include asymptomatic individuals using pooled saliva.

One Sentence Summary: GTR-STM and Direct-into-PCR GTR-STMdk offer substantive improvements in SARS-CoV-2 viral RNA stability, safety, and RT-PCR process efficiency for COVID-19 testing by using a non-dilutive saliva sample collection system for individuals at home or onsite respectively.
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http://dx.doi.org/10.1101/2021.01.20.20243782DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7852251PMC
January 2021

Research Note: Phytobiotics modulate the expression profile of circulating inflammasome and cyto(chemo)kine in whole blood of broilers exposed to cyclic heat stress.

Poult Sci 2021 Mar 5;100(3):100801. Epub 2020 Nov 5.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville 72701, USA. Electronic address:

Heat stress (HS) is a critical concern to the poultry industry as it affects both productivity and well-being. Various managerial and nutritional strategies have been proposed to mitigate the negative effects of HS in chickens, with plant-based additives showing promise. Recently, we reported the positive effect of a phytogenic feed additive (PFA) on growth performance in HS birds. Owing to the antioxidant nature of these compounds, we sought to further explore the effect of PFA on whole blood circulating chemokines, cytokines, and inflammasomes in HS broilers. Broilers (600 males, 1 d) were randomly assigned to 12 environmental chambers, subjected to 2 environmental conditions (12 h cyclic heat stress, HS, 35°C vs. thermoneutral condition [TN], 24°C) and fed 3 diets (control, PFA-C 250 ppm, PFA-C 400 ppm) in a 2 × 3 factorial design. After 21 d of cyclic HS, blood samples were collected for target gene expression analysis. HS upregulated the expression of superoxide dismutase 1 (SOD1) and downregulated glutathione peroxidase-3 (GPX-3), and there was diet × temperature interaction for SOD2, GPX-1, and GPX-3, where gene expression was increased by PFA-C250 during HS but was unchanged for PFA-C400. Plasma total antioxidant capacity (TAC) and malondialdehyde (MDA) content were increased by HS. Gene expression of interleukin-18 (IL-18) was decreased by HS, without further effect of PFA. HS increased tumor necrosis factor α (TNFα), but this effect was mitigated by PFA-C400. C-C motif chemokine ligands 4 and 20 (CCL4 and CCL20) showed a similar pattern to TNFα, with PFA-C400 ameliorating the negative effect of HS. The nucleotide-binding, leucine-rich repeat and pyrin domain containing 3 (NLRP3) inflammasome was decreased by HS and further lowered by PFA-C400, but the nucleotide-binding oligomerization domain, leucine-rich repeat, and CARD domain containing 3 (NLRC3) and nucleotide-binding, leucine-rich repeat containing X1 (NLRX1) inflammasomes were increased by PFA under TN conditions, with no effects of HS. Heat shock proteins (HSP) and heat shock factors (HSF) were unaffected by PFA or HS. Together these data indicate that gene expression of circulating inflammatory factors are dysregulated during HS, and supplemental dietary PFA may be protective.
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http://dx.doi.org/10.1016/j.psj.2020.10.055DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7936152PMC
March 2021

Optimization of a transient antibody expression platform towards high titer and efficiency.

Biotechnol J 2021 Apr 25;16(4):e2000251. Epub 2021 Jan 25.

Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, Connecticut, USA.

Transient gene expression (TGE) using mammalian cells is an extensively used technology for the production of antibodies and recombinant proteins and has been widely adopted by both academic and industrial labs. Chinese Hamster Ovary (CHO) cells have become one of the major workhorses for TGE of recombinant antibodies due to their attractive features: post-translational modifications, adaptation to high cell densities, and use of serum-free media. In this study, we describe the optimization of parameters for TGE for antibodies from CHO cells. Through a matrix evaluation of multiple factors including inoculum, transfection conditions, amount and type of DNA used, and post-transfection culture conditions, we arrived at an uniquely optimized process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply. We further investigated the amount of coding DNA used in TGE and the influence of kinetics and size of the transfection complex on the in vitro efficiency of the transfection. We present here the first report of an optimized TGE platform using Filler DNA in an early drug discovery setting for the screening and production of therapeutic mAbs.
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http://dx.doi.org/10.1002/biot.202000251DOI Listing
April 2021

Muscle Metabolome Profiles in Woody Breast-(un)Affected Broilers: Effects of Quantum Blue Phytase-Enriched Diet.

Front Vet Sci 2020 4;7:458. Epub 2020 Aug 4.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR, United States.

Woody breast (WB) myopathy is significantly impacting modern broilers and is imposing a huge economic burden on the poultry industry worldwide. Yet, its etiology is not fully defined. In a previous study, we have shown that hypoxia and the activation of its upstream mediators (AKT/PI3K/mTOR) played a key role in WB myopathy, and supplementation of quantum blue (QB) can help to reduce WB severity via modulation of hypoxia-related pathways. To gain further insights, we undertook here a metabolomics approach to identify key metabolite signatures and outline their most enriched biological functions. Ultra performance liquid chromatography coupled with high resolution mass spectrometry (UPLC-HRMS) identified a total of 108 known metabolites. Of these, mean intensity differences at < 0.05 were found in 60 metabolites with 42 higher and 18 lower in WB-affected compared to unaffected muscles. Multivariate analysis and Partial Least Squares Discriminant analysis (PLS-DA) scores plot displayed different clusters when comparing metabolites profile from affected and unaffected tissues and from moderate (MOD) and severe (SEV) WB muscles indicating that unique metabolite profiles are present for the WB-affected and unaffected muscles. To gain biologically related molecule networks, a stringent pathway analyses was conducted using IPA knowledge-base. The top 10 canonical pathways generated, using a fold-change -1.5 and 1.5 cutoff, with the 50 differentially abundant-metabolites were purine nucleotide degradation and biosynthesis, sirtuin signaling pathway, citrulline-nitric oxide cycle, salvage pathways of pyrimidine DNA, IL-1 signaling, iNOS, Angiogenesis, PI3K/AKT signaling, and oxidative phosphorylation. The top altered bio-functions in term of molecular and cellular functions in WB-affected tissues included cellular development, cellular growth and proliferation, cellular death and survival, small molecular biochemistry, inflammatory response, free radical scavenging, cell signaling and cell-to-cell interaction, cell cycles, and lipid, carbohydrate, amino acid, and nucleic acid metabolisms. The top disorder functions identified were organismal injury and abnormalities, cancer, skeletal and muscular disorders, connective tissue disorders, and inflammatory diseases. Breast tissues from birds fed with high dose (2,000 FTU) of QB phytase exhibited 22 metabolites with significantly different levels compared to the control group with a clear cluster using PLS-DA analysis. Of these 22 metabolites, 9 were differentially abundant between WB-affected and unaffected muscles. Taken together, this study determined many metabolic signatures and disordered pathways, which could be regarded as new routes for discovering potential mechanisms of WB myopathy.
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http://dx.doi.org/10.3389/fvets.2020.00458DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7417653PMC
August 2020

Vitrification of Heart Valve Tissues.

Methods Mol Biol 2021 ;2180:593-605

Tissue Testing Technologies LLC, North Charleston, SC, USA.

Application of the original vitrification protocol used for pieces of heart valves to intact heart valves has evolved over time. Ice-free cryopreservation by Protocol 1 using VS55 is limited to small samples (1-3 mL total volume) where relatively rapid cooling and warming rates are possible. VS55 cryopreservation typically provides extracellular matrix preservation with approximately 80% cell viability and tissue function compared with fresh untreated tissues. In contrast, ice-free cryopreservation using VS83, Protocols 2 and 3, permits preservation of large samples (80-100 mL total volume) with several advantages over conventional cryopreservation methods and VS55 preservation, including long-term preservation capability at -80 °C; better matrix preservation than freezing with retention of material properties; very low cell viability, reducing the risks of an immune reaction in vivo; reduced risks of microbial contamination associated with use of liquid nitrogen; improved in vivo functions; no significant recipient allogeneic immune response; simplified manufacturing process; increased operator safety because liquid nitrogen is not used; and reduced manufacturing costs. More recently, we have developed Protocol 4 in which VS55 is supplemented with sugars resulting in reduced concerns regarding nucleation during cooling and warming. This method can be used for large samples resulting in retention of cell viability and permits short-term exposure to -80 °C with long-term storage preferred at or below -135 °C.
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http://dx.doi.org/10.1007/978-1-0716-0783-1_31DOI Listing
March 2021

Research Note: Dietary phytase reduces broiler woody breast severity via potential modulation of breast muscle fatty acid profiles.

Poult Sci 2020 Aug 15;99(8):4009-4015. Epub 2020 Jun 15.

Center of Excellence for Poultry Science, Division of Agriculture, University of Arkansas, Fayetteville 72701. Electronic address:

Woody breast (WB) myopathy is a major concern and economic burden to the poultry industry, and for which, there is no effective solution because of its unknown etiology. In a previous study, we have shown that phytase (Quantum Blue, QB) reduces the WB severity by 5% via modulation of oxygen homeostasis-related pathways. As WB has been suggested to be associated with lipid dysmetabolism, we aimed to determine the effect of QB on WB and breast muscle fatty acid profile. Male broilers were subjected to 6 treatments (96 birds/treatment): a nutrient adequate control group (PC), the PC supplemented with 0.3% myo-inositol (PC + MI), a negative control (NC) deficient in available P and Ca by 0.15 and 0.16%, respectively, the NC fed with QB at 500 (NC+500 FTU), and 1,000 (NC+ 1,000 FTU) or 2,000 FTU/kg of feed (NC+2,000 FTU). Woody breast and white striping scores were recorded, and fatty acid profiles were determined using gas liquid chromatography. Woody breast-affected muscles exhibited a significant higher incidence of white striping as liquid chromatography analysis reveals an imbalance of fatty acid profile in the breast of WB-affected birds with a significant higher percent of saturated fatty acids (SFA, myristic [14:0], pentadecanoic [15:0], and margaric [17:0]) and monounsaturated fatty acids (myristoleic [14:1], palmitoleic [16:1c], 10-trans-heptadecenoic [17:1t], oleic [18:1c9], and vaccenic [18:1c11]), and lower content of polyunsaturated fatty acids (PUFA) and omega-3 (P < 0.05). Quantum Blue at high doses (1,000 and 2,000 FTU) significantly reduces the percent of SFA and increases that of PUFA compared with the control group. In conclusion, WB myopathy seemed to be associated with an imbalance of fatty acid profile, and QB ameliorates the severity of WB potentially via modulation of SFA and PUFA contents.
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http://dx.doi.org/10.1016/j.psj.2020.05.005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7597982PMC
August 2020

Extension Horses, Inc. experts act fast to create online resources to assist the horse industry during COVID-19.

Transl Anim Sci 2020 Jul 25;4(3):txaa085. Epub 2020 Jun 25.

Pennsylvania State University, Department of Animal Science, University Park, PA.

The COVID-19 pandemic and resulting stay-at-home directives, adopted out of necessity to protect human health, introduced significant challenges for horse owners and small equine businesses. Restricted access, and in many cases closure of barns, resulted in a multitude of questions and concerns within the equine community which needed to be addressed rapidly. Extension Horses, Inc. (EH) coordinated the development and delivery of a variety of educational resources utilizing a combination of online formats and dissemination through social media and EH member contact lists. A series of infographics, webinars, and podcasts (three in each category) were created to provide guidance on essential care of horses, emergency preparedness, financial assistance, legal concerns, and biosecurity during the crucial, initial weeks of the pandemic (March to April 2020). Web conferencing technology (Zoom) was used to facilitate discussion and task delegation among EH members and to conduct and record webinars and podcasts. Podcasts were hosted on Buzzsprout and infographics were created using Adobe InDesign. Live webinar participants were invited to participate in several polls during the webinar and were sent a brief survey to complete at the end of the webinar series. Analytics for all educational resources combined demonstrated a 32-d total direct reach of 135,563. Most live webinar participants identified themselves as horse owners and small equine business owners (55%). The majority of live webinar participants indicated the information was useful (99%), and they would utilize the resources they had learned about (80%). Survey respondents reported that Facebook, email, and word of mouth were key ways in which they learned about the webinars. The same survey found that the web-platform was an effective method to receive information (85% high satisfaction) and respondents were highly likely to recommend future EH webinars to others (88%). The three infographics had a total Facebook reach of 131,765, the webinars had 3,522 total views, and the three podcasts had 276 total downloads. The rapid response of EH and quick turnaround of products allowed a large online audience to receive vital information for coping with COVID-19. Having the established EH network, already familiar with virtual education, was a big asset in this effort. This can serve as a model for cooperative extension to utilize in future collaborative responses to industry issues.
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http://dx.doi.org/10.1093/tas/txaa085DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7337838PMC
July 2020

Monitoring Phytate Hydrolysis Using Serial Blood Sampling and Feather Myo-Inositol Levels in Broilers.

Front Physiol 2020 26;11:736. Epub 2020 Jun 26.

Department of Poultry Science, University of Arkansas, Fayetteville, AR, United States.

Phytate forms insoluble precipitates with various cations that are recalcitrant to digestion in poultry. Dietary supplementation with exogenous phytase has been shown to improve phytate solubility and digestibility and, in turn, improve animal growth performance. Although the kinetics of phytate hydrolysis by exogenous phytase are well described , the progression of the reaction is still not well defined. The aim of the present study was, therefore, to monitor the kinetic variation of myo-inositol (myo-Ins) levels in both circulation and feather following exogenous phytase supplementation. In experiment 1, 4 week-old male broilers were individually housed with access to water and a standard commercial diet. Birds were maintained under environmental temperature of 24°C and 30% RH. Birds were cannulated in the cutaneous ulnar vein on the right wing and remained untouched for 3 days. On the day of the experiment, birds were randomly divided into three body weight-matched groups and fed either the control diet, the control diet-supplemented with myo-Ins or Ronozyme HiPhos (0.06%, DSM Nutritional Products, Switzerland) for 10 h. In the experiment 2, birds were fed only HiPhos for 30 h. Growing feathers and blood were collected at baseline and then every 2 h for 10 h (experiment 1) and 30 h (experiment 2) post-prandially. Plasma and feather myo-Ins levels were determined by UHPLC-MS/MS. The relative expression of inositol polyphosphate-1-phosphatase (INPP1), inositol hexakisphosphate kinase 1-3 (IP6K1-3), inositol-3-phosphate synthase (ISYNA), and multiple inositol-polyphosphate phosphatase 1 (MNPP1) genes in blood and feathers was determined by real-time qPCR using 2 method. Plasma and feather myo-Ins levels were significantly increased by HiPhos at 6 h to 8 h post-prandial. The mRNA abundances of INPP1, IP6K1, and ISYNA in the circulation were significantly down regulated at all periods compared to the baseline levels. IP6K2, IP6K3, and MINPP1 gene expression, however, was up regulated at 8 h post-prandial and then returned to the baseline levels. In feathers, the expression of INPP1 was induced at 8 h post-prandial and remained higher compared to the baseline. The expression of IP6K2, IP6K3, and MINPP1 was down regulated during the first 10 h and then returned to baseline levels for the rest of the post-prandial period. Taken together, our data show that phytase modulates the expression of genes associated with myo-Ins metabolism and generates release of myo-Ins in both circulation and feather at 6-10 h post-feeding. Feather myo-Ins concentration could be used as a non-invasive method to monitor phytate hydrolysis in practice.
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http://dx.doi.org/10.3389/fphys.2020.00736DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7333251PMC
June 2020

Intestinal Barrier Integrity in Heat-Stressed Modern Broilers and Their Ancestor Wild Jungle Fowl.

Front Vet Sci 2020 7;7:249. Epub 2020 May 7.

Department of Poultry Science, University of Arkansas, Fayetteville, AR, United States.

High environmental temperature has strong adverse effects on poultry production, welfare, and sustainability and, thereby, constitutes one of the most challenging stressors. Although colossal information has been published on the effects of heat stress on poultry productivity and gut health, the fundamemntal mechanisms associated with heat stress responses and intestinal barrier function are still not well defined. The aim of the present study was, therefore, to determine the effects of acute (2 h) heat stress on growth performance, gut integrity, and intestinal expression of heat shock and tight junction proteins in slow- (broilers of the 1950's, ACRB), moderate- (broilers of 1990's, 95RAN), rapid-(modern broilers, MRB) growing birds, and their ancestor wild jungle fowl (JF). Heat stress exposure significantly increased the core body temperature of 95RAN and MRB chickens by ~0.5-1°C, but not that of JF and ACRB compared to their counterparts maintained at thermoneutral conditions. Heat stress also depressed feed intake and increased serum fluorescein isothiocyanate-dextran (FITC-D) levels ( < 0.05) in modern broilers (95RAN and MRB) but not in JF and ACRB, indicating potential leaky gut syndrome. Molecular analyses showed that heat stress exposure significantly up regulated the duodenal expression of occludin ( and lipocalin ( in ACRB, zonula occludens (, villin1 , and calprotectin ( in 95 RAN, and only in MRB compared to their TN counterparts. In the jejunum however, heat stress down regulated the expression of PALS1-associated tight junction protein ( in ACRB, 95RAN, and MRB, and that of cadherin1 ( in MRB. In the ileum, heat stress significantly down regulated the expression of in 95 RAN, in MRB, gap junction protein alpha1 ( in JF, and in ACRB compared to their TN counterparts. In summary, this is the first report, to our knowledge, showing that tight junction protein expression is environmental-, genotype-, and intestinal segment-dependent and identifying molecular signatures, such as , and , potentially involved in leaky gut syndrome-induced by heat stress in MRB.
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http://dx.doi.org/10.3389/fvets.2020.00249DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7220999PMC
May 2020

Research Note: Evaluation of a heat stress model to induce gastrointestinal leakage in broiler chickens.

Poult Sci 2020 Mar 27;99(3):1687-1692. Epub 2019 Dec 27.

Department of Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA. Electronic address:

The purpose of this study was to evaluate heat stress as a model to induce gastrointestinal leakage in broiler chickens. On the day of hatch, 320 chicks were allocated into 8 environmental chambers, 4 thermoneutral (TN) and 4 continuous heat stress (HS). Each chamber was divided into 2 pens containing separate feeders and water jugs (8 replicates per treatment, 20 birds/pen). The environment was established to simulate production setting as best possible for the first 21 D. A gradual reduction of temperature from 32°C to 24°C with relative humidity at 55 ± 5% was adopted for the first 21 D. At the time of HS, the HS groups were exposed to 35°C from Day 21 to 42, while thermoneutral ones were maintained at 24°C from Day 21 to 42. Chickens were equipped with a Thermochron temperature logger for continuous monitoring of core body temperature. The environmental temperature and relative humidity were continuously recorded. Fluorescein isothiocyanate-dextran (FITC-d) was orally gavaged to 2 chickens/replicate (n = 16) randomly selected on days 21, 28, 35, and 42. After 1 h of oral gavage, blood samples were collected to determine the passage of FITC-d. Tibias were removed from all chickens to evaluate break strength only on 21 D and 42 D (before HS and at the end of the trial). Performance parameters were evaluated weekly from 21 D to the end of the trial. Body temperature was significantly (P < 0.05) increased after 2 h of starting HS and remained that way until the end of the study. Chronic HS caused an increase in core body temperature which decreased feed intake, body weight, and feed efficiency (28, 35, and 42 D) when compared with control TN chickens. Similarly, serum FITC-d was significantly increased in HS chickens at all points of evaluation. Chronic HS also caused a significant reduction of bone strength at 42 D when compared with the control chickens. The results from the present study suggest that HS can be a robust model to induce gut leakage in broiler chickens.
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http://dx.doi.org/10.1016/j.psj.2019.10.075DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7587818PMC
March 2020

Water amino acid-chelated trace mineral supplementation decreases circulating and intestinal HSP70 and proinflammatory cytokine gene expression in heat-stressed broiler chickens.

J Anim Sci 2020 Mar;98(3)

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR.

Heat stress (HS) is a financial and physiological burden on the poultry industry and the mitigation of the adverse effects of HS is vital to poultry production sustainability. The purpose of this study was, therefore, to determine the effects of an amino acid-chelated trace mineral supplement on growth performance, stress and inflammatory markers, and meat quality in heat-stressed broilers. One day-old Cobb 500 male broilers (n = 480) were allocated into 12 environmental chambers (24 floor pens) and divided into two groups: one group supplemented with amino acid-chelated trace mineral in drinking water and one control group. On day 28, birds were subjected to chronic heat stress (HS, 2 wk, 35 °C and 20% to 30% RH) or maintained at thermoneutral condition (TN, 24 °C) in a 2 × 2 factorial design. Feed intake (FI), water consumption, and body weight were recorded. At day 42, serum fluorescein isothiocyanate dextran (FITC-D) levels, blood gas, electrolyte, and stress markers were measured. Jejunum samples were collected to measure gene expression of stress, inflammation, and tight junction proteins. The rest of the birds were processed to evaluate carcass traits. HS resulted in an increase in core body temperature, which increased water intake and decreased FI, body weight, and feed efficiency (P < 0.05). HS reduced carcass yield and the weight of all parts (P < 0.05). HS significantly increased levels of circulating corticosterone (CORT), heat shock protein 70 (HSP70), interleukin 18 (IL-18), tumor necrosis factor alpha, C-reactive protein, and nucleotide-binding oligomerization domain leucine-rich repeat and pyrin domain-containing 3 expression. HS significantly increased serum FITC-D levels and the expression of HSP70 and IL-18 in the jejunum. Although it did not affect the growth performance, amino acid-chelated trace mineral supplementation reversed the effect of HS by reducing CORT and FITC-D levels and the expression of stress and proinflammatory cytokines in the circulation and the jejunum. However, it upregulated these parameters in birds maintained under TN conditions. Together, these data indicate that the amino acid-chelated trace mineral might alleviate stress and inflammation and improve gut integrity in heat-stressed but not thermoneutral broilers.
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http://dx.doi.org/10.1093/jas/skaa049DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7070152PMC
March 2020

Phytogenic feed- and water-additives improve feed efficiency in broilers via modulation of (an)orexigenic hypothalamic neuropeptide expression.

Neuropeptides 2020 Jun 3;81:102005. Epub 2020 Jan 3.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR 72701, United States of America. Electronic address:

Fueled by consumer preference for natural and antibiotic-free products, phytogenics have become the fastest growing segment of the animal feed additives. Yet, their modes of action are not fully understood. This study was undertaken to determine the effect of 5 phytogenics (3 feed- and 2 water-supplements) on the growth performance of commercial broilers, and their potential underlying molecular mechanisms. Day-old male Cobb 500 chicks (n = 576) were randomly assigned into 48 pens consisting of 6 treatments (Control; AVHGP; SCP; BHGP; AVSSL; SG) in a complete randomized design (12 birds/pen, 8 pens/treatment, 96 birds/treatment). Chicks had ad libitum access to feed and water. Individual body weight (BW) was recorded weekly and feed intake was measured daily. Core body temperatures were continuously recorded using thermo-loggers. At d 35, hypothalamic tissues were excised from the thermo-logger-equipped chickens (n = 8 birds/treatment) to determine the expression of feeding-related neuropeptides. Both feed (AVHGP, SCP, BHGP) and water-supplemented (AVSSL, SG) phytogenics significantly improved feed efficiency (FE) compared to the control birds. This higher FE was achieved via a reduction in core body temperature and improvement of market BW, without changes in feed intake in broilers supplemented with phytogenic water additives as compared to the control group. Broilers fed dietary phytogenics, however, attained higher feed efficiency via a reduction in feed intake while maintaining similar BW as the control group. At the molecular levels, the effects of the phytogenic water additives seemed to be mediated by the activation of the hypothalamic AgRP-ORX-mTOR-S6k1 and inhibition of CRH pathways. The effect of the phytogenic feed additives appeared to be exerted through the activation of AdipoQ, STAT3, AMPK, and MC1R pathways. This is the first report describing the likely central mechanisms through which phytogenic additives improve the growth performance and feed efficiency in broilers.
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http://dx.doi.org/10.1016/j.npep.2020.102005DOI Listing
June 2020

Imaging the distribution of iron oxide nanoparticles in hypothermic perfused tissues.

Magn Reson Med 2020 05 9;83(5):1750-1759. Epub 2019 Dec 9.

Center for Magnetic Resonance Research, Department of Radiology, University of Minnesota, Minneapolis, Minnesota.

Purpose: Herein, we evaluate the use of MRI as a tool for assessing iron oxide nanoparticle (IONP) distribution within IONP perfused organs and vascularized composite allografts (VCAs) (i.e., hindlimbs) prepared for cryopreservation.

Methods: Magnetic resonance imaging was performed on room-temperature organs and VCAs perfused with IONPs and were assessed at 9.4 T. Quantitative T mapping and -weighted images were acquired using sweep imaging with Fourier transformation and gradient-echo sequences, respectively. Verification of IONP localization was performed through histological assessment and microcomputer tomography.

Results: Quantitative imaging was achieved for organs and VCAs perfused with up to 642 mM (36 mg /mL), which is above previous demonstrations of upper limit detection in agarose (35.7mM [2 mg /mL]). The stability of IONPs in the perfusate had an effect on the quality of distribution and imaging within organs or VCA. Finally, MRI provided more accurate IONP localization than Prussian blue histological staining in this system, wherein IONPs remain primarily in the vasculature.

Conclusion: Using MRI, we were able to assess the distribution of IONPs throughout organs and VCAs varying in complexity. Additional studies are necessary to better understand this system and validate the calibration between T measurements and IONP concentration.
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http://dx.doi.org/10.1002/mrm.28123DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982566PMC
May 2020

75-kDa glucose-regulated protein (GRP75) is a novel molecular signature for heat stress response in avian species.

Am J Physiol Cell Physiol 2020 02 4;318(2):C289-C303. Epub 2019 Dec 4.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, Arkansas.

Glucose-regulated protein 75 (GRP75) was first characterized in mammals as a heat shock protein-70 (HSP70) family stress chaperone based on its sequence homology. Extensive studies in mammals showed that GRP75 is induced by various stressors such as glucose deprivation, oxidative stress, and hypoxia, although it remained unresponsive to the heat shock. Such investigations are scarce in avian (nonmammalian) species. We here identified chicken GRP75 by using immunoprecipitation assay integrated with LC-MS/MS, and found that its amino acid sequence is conserved with high homology (52.5%) to the HSP70 family. Bioinformatics and 3D-structure prediction indicate that, like most HSPs, chicken GRP75 has two principal domains (the NH-terminal ATPase and COOH-terminal region). Immunofluorescence staining shows that GRP75 is localized predominantly in the avian myoblast and hepatocyte mitochondria. Heat stress exposure upregulates GRP75 expression in a species-, genotype-, and tissue-specific manner. Overexpression of GRP75 reduces avian cell viability, and blockade of GRP75 by its small molecular inhibitor MKT-077 rescues avian cell viability during heat stress. Taken together, this is the first evidence showing that chicken GRP75, unlike its mammalian ortholog, is responsive to heat shock and plays a key role in cell survival/death pathways. Since modern avian species have high metabolic rates and are sensitive to high environmental temperature, GRP75 could open new vistas in mechanistic understanding of heat stress responses and thermotolerance in avian species.
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http://dx.doi.org/10.1152/ajpcell.00334.2019DOI Listing
February 2020

Comparison of liver gene expression by RNAseq and PCR analysis after 8 weeks of feeding soy protein isolate- or casein-based diets in an obese liver steatosis rat model.

Food Funct 2019 Dec;10(12):8218-8229

Department of Poultry Science & The Center of Excellence for Poultry Science, University of Arkansas, 1260 W. Maple, Fayetteville, AR 72701, USA.

Previously, we reported that feeding soy protein isolate (SPI) reduced liver steatosis in obese rats compared to those fed a casein (CAS)-based diet; however, the mechanism for this protection is unknown. To gain insight into the ability of SPI to ameliorate liver steatosis, we conducted transcriptomic (RNAseq) analysis on liver samples from obese rats fed either the SPI- or CAS-based diets (n = 8 per group) for 8 weeks using an Ilumina HiSeq with 100 base paired end reads for sequencing. Data were analyzed by Ingenuity Pathway Analysis (IPA) software using a P < 0.05 and 1.3-fold differential expression cutoff values between the SPI- and CAS-based groups. To independently validate the RNAseq data, we conducted targeted mRNA expression analysis using quantitative PCR (qPCR) on a subset of differently expressed genes. The results indicate that mRNA expression by qPCR concurred with RNAseq for NPTX2, GPT, INMT, and HAL that were up-regulated in SPI-fed rats (P < 0.05) and PRSS8, AJUBA, CSF2RB, and Cyp2c12 that were down-regulated (P < 0.05) in SPI-fed rats compared to CAS-fed rats. Our findings may shed light on understanding mechanisms enabling SPI diet to reduce liver steatosis in this obese Zucker rat model.
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http://dx.doi.org/10.1039/c9fo01387cDOI Listing
December 2019

Hormonal regulation of visfatin gene in avian Leghorn male hepatoma (LMH) cells.

Comp Biochem Physiol A Mol Integr Physiol 2020 02 24;240:110592. Epub 2019 Oct 24.

University of Arkansas, Center of Excellence for Poultry Science, Fayetteville, AR 72701, USA. Electronic address:

Visfain has been extensively studied in mammals and has been shown to play an important role in obesity and insulin resistance. However, there is a paucity of information on visfatin regulation in non-mammalian species. After characterization of chicken visfatin gene, we undertook this study to determine its hormonal regulation in avian (non-mammalian) liver cells. Addition of 5 ng/mL TNFα, 100 ng/mL leptin, 1, 3, 10 or 100 ng/mL T3 for 24 h upregulated visfatin gene expression by 1.2, 1.8, 1.95, 1.75, 1.80, and 2.45 folds (P < .05), respectively, compared to untreated LMH cells. Administration of 10 ng/mL of orexin A significantly down regulated visfatin gene expression by 1.35 folds compared to control cells. In contrast, treatment with IL-6 or orexin B for 24 h did not influence visfatin mRNA abundance. These pro-inflammatory cytokines and obesity-related hormones modulate the expression of CRP, INSIG2, and nuclear orphan receptors. Hepatic CRP gene expression was significantly upregulated by IL-6, TNFα, orexin B, and T3 and down regulated by leptin and orexin A. LXR mRNA abundances were increased by orexin A, decreased by orexin B, and T3, and did not affected by IL6, TNFα, or leptin. The expression of FXR gene was induced by IL-6, leptin, and T3, but it was not influenced by TNFα, orexin A or B. CXR gene expression was up regulated by TNFα, leptin, orexin B, and T3, down regulated by 5 ng/mL orexin A, and did not affected by IL-6. INSIG2 mRNA levels were increased by TNFα (5 ng/mL), leptin (100 ng/mL), and T3 (1, 3, 10, and 100 ng/mL), decreased by orexin A, and remained unchanged with IL-6 or orexin B treatment. Together, this is the first report showing hormonal regulation of visfatin in avian hepatocyte cells and suggesting a potential role of CRP, INSIG2, and nuclear orphan receptor LXR, FXR, and CXR in mediating these hormonal effects.
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http://dx.doi.org/10.1016/j.cbpa.2019.110592DOI Listing
February 2020

Novel Autoimmune IgM Antibody Attenuates Atherosclerosis in IgM Deficient Low-Fat Diet-Fed, but Not Western Diet-Fed Mice.

Arterioscler Thromb Vasc Biol 2020 01 24;40(1):206-219. Epub 2019 Oct 24.

From the Robert M. Berne Cardiovascular Research Center (O.A.C., P.S., E.S.G., R.M.H., M.E.M., N.L., C.A.M., G.K.O.), University of Virginia, Charlottesville.

Objective: Oxidized phospholipids (OxPL), such as the oxidized derivatives of 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine, 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine, and 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, have been shown to be the principal biologically active components of minimally oxidized LDL (low-density lipoprotein). The role of OxPL in cardiovascular diseases is well recognized, including activation of inflammation within vascular cells. Atherosclerotic mice fed a high-fat diet develop antibodies to OxPL, and hybridoma B-cell lines producing natural anti-OxPL autoantibodies have been successfully generated and characterized. However, as yet, no studies have been reported demonstrating that treatment with OxPL neutralizing antibodies can be used to prevent or reverse advanced atherosclerosis. Approach and Results: Here, using a screening against 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine/1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine, we generated a novel IgM autoantibody, 10C12, from the spleens of mice fed a long-term Western diet, that demonstrated potent OxPL neutralizing activity in vitro and the ability to inhibit macrophage accumulation within arteries of mice fed a Western diet for 4 weeks. Of interest, 10C12 failed to inhibit atherosclerosis progression in mice treated between 18 and 26 weeks of Western diet feeding likely due at least in part to high levels of endogenous anti-OxPL antibodies. However, 10C12 treatment caused a 40% decrease in lipid accumulation within aortas of secreted IgM deficient, , mice fed a low-fat diet, when the antibody was administrated between 32-40 weeks of age.

Conclusions: Taken together, these results provide direct evidence showing that treatment with a single autoimmune anti-OxPL IgM antibody during advanced disease stages can have an atheroprotective outcome.
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http://dx.doi.org/10.1161/ATVBAHA.119.312771DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7006879PMC
January 2020

Quantum Blue Reduces the Severity of Woody Breast Myopathy via Modulation of Oxygen Homeostasis-Related Genes in Broiler Chickens.

Front Physiol 2019 1;10:1251. Epub 2019 Oct 1.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR, United States.

The incidence of woody breast (WB) is increasing on a global scale representing a significant welfare problem and economic burden to the poultry industry and for which there is no effective treatment due to its unknown etiology. In this study, using diffuse reflectance spectroscopy (DRS) coupled with iSTAT portable clinical analyzer, we provide evidence that the circulatory- and breast muscle-oxygen homeostasis is dysregulated [low oxygen and hemoglobin (HB) levels] in chickens with WB myopathy compared to healthy counterparts. Molecular analysis showed that blood HB subunit Mu (HBM), Zeta (HBZ), and hephaestin (HEPH) expression were significantly down regulated; however, the expression of the subunit rho of HB beta (HBBR) was upregulated in chicken with WB compared to healthy counterparts. The breast muscle HBBR, HBE, HBZ, and hypoxia-inducible factor prolyl hydroxylase 2 (PHD2) mRNA abundances were significantly down regulated in WB-affected compared to normal birds. The expression of HIF-1α at mRNA and protein levels was significantly induced in breasts of WB-affected compared to unaffected birds confirming a local hypoxic status. The phosphorylated levels of the upstream mediators AKT at Ser473 site, mTOR at Ser2481 site, and PI3K P85 at Tyr458 site, as well as their mRNA levels were significantly increased in breasts of WB-affected birds. In attempt to identify a nutritional strategy to reduce WB incidence, male broiler chicks (Cobb 500, = 576) were randomly distributed into 48 floor pens and subjected to six treatments (12 birds/pen; 8 pens/treatment): a nutrient adequate control group (PC), the PC supplemented with 0.3% myo-inositol (PC + MI), a negative control (NC) deficient in available P and Ca by 0.15 and 0.16%, respectively, the NC fed with quantum blue (QB) at 500 (NC + 500 FTU), 1,000 (NC + 1,000 FTU), or 2,000 FTU/kg of feed (NC + 2,000 FTU). Although QB-enriched diets did not affect growth performances (FCR and FE), it did reduce the severity of WB by 5% compared to the PC diet. This effect is mediated by reversing the expression profile of oxygen homeostasis-related genes; i.e., significant down regulation of HBBR and upregulation of HBM, HBZ, and HEPH in blood, as well as a significant upregulation of HBA1, HBBR, HBE, HBZ, and PHD2 in breast muscle compared to the positive control.
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http://dx.doi.org/10.3389/fphys.2019.01251DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6781743PMC
October 2019

Double-Stranded RNA Is a Novel Molecular Target in Osteomyelitis Pathogenesis: A Translational Avian Model for Human Bacterial Chondronecrosis with Osteomyelitis.

Am J Pathol 2019 10 2;189(10):2077-2089. Epub 2019 Aug 2.

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, Arkansas; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, Arkansas. Electronic address:

Osteomyelitis remains a serious inflammatory bone disease that affects millions of individuals worldwide and for which there is no effective treatment. Despite scientific evidence that Staphylococcus bacteria are the most common causative species for human bacterial chondronecrosis with osteomyelitis (BCO), much remains to be understood about the underlying virulence mechanisms. Herein, we show increased levels of double-stranded RNA (dsRNA) in infected bone in a Staphylococcus-induced chicken BCO model and in human osteomyelitis samples. Administration of synthetic [poly(I:C)] or genetic (Alu) dsRNA induces human osteoblast cell death. Similarly, infection with Staphylococcus isolated from chicken BCO induces dsRNA accumulation and cell death in human osteoblast cell cultures. Both dsRNA administration and Staphylococcus infection activate NACHT, LRR and PYD domains-containing protein (NLRP)3 inflammasome and increase IL18 and IL1B gene expression in human osteoblasts. Pharmacologic inhibition with Ac-YVAD-cmk of caspase 1, a critical component of the NLRP3 inflammasome, prevents DICER1 dysregulation- and dsRNA-induced osteoblast cell death. NLRP3 inflammasome and its components are also activated in bone from BCO chickens and humans with osteomyelitis, compared with their healthy counterparts. These findings provide a rationale for the use of chicken BCO as a human-relevant spontaneous animal model for osteomyelitis and identify dsRNA as a new treatment target for this debilitating bone pathogenesis.
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http://dx.doi.org/10.1016/j.ajpath.2019.06.013DOI Listing
October 2019

Evaluation of Bone Marrow Adipose Tissue and Bone Mineralization on Broiler Chickens Affected by Wooden Breast Myopathy.

Front Physiol 2019 29;10:674. Epub 2019 May 29.

Department of Poultry Science, University of Arkansas, Fayetteville, AR, United States.

In humans, alterations in bone metabolism have been associated with myopathies. We postulate the hypothesis that perhaps similar pathologies can also be associated in modern chickens. Hence, this study aimed to assess the fat infiltration in bone marrow and its repercussion on broiler chicken affected by Wooden Breast (WB) myopathy. Ten Cobb 500 live birds with extreme rigidity of the (PM) muscle were selected as WB affected chickens by physical examination of the muscle at 49 days of age, whereas ten chickens healthy with no physical signs of hardness in the breast muscle were considered to be unaffected. Macroscopic lesions in affected chickens included areas of firm and inflamed muscle with pale appearance, hemorrhaging, and viscous exudate on the surface. Bone marrow and sections of the PM muscle were collected and analyzed for light microscopy. Additionally, transmission electron microscopy was conducted in affected or unaffected muscle. Chickens affected with WB showed significant reductions ( < 0.05) in femur diameter, calcium, and phosphorous percentage but increased breast weight, compression force and filet thickness when compared with non-affected chickens. Interestingly, bone marrow from WB chicken had subjectively, more abundant infiltration of adipose tissue, when compared with non-affected chickens. Histology of the Pectoralis major of birds with WB showed abundant infiltration of adipose tissue, muscle fibers degeneration with necrosis and infiltration of heterophils and mononuclear cells, connective tissue proliferation, and vasculitis. Ultrastructural changes of WB muscle revealed lack definition of bands in muscle tissue, or any normal ultrastructural anatomy such as myofibrils. The endomysium components were necrotic, and in some areas, the endomysium was notable only as a string of necrotic tissue between degraded myofibrils. The fascia appeared hypertrophied, with large areas of necrosis and myofiber without structural identity with degraded mitochondria adjacent to the disrupted muscle tissue. As far as we know, this is the first study that describes a subjective increase in adipose tissue in the bone marrow of chickens affected with WB when compared with non-affected chickens, and reduced bone mineralization.
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http://dx.doi.org/10.3389/fphys.2019.00674DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6549442PMC
May 2019

Evaluation of Intestinal Permeability and Liver Bacterial Translocation in Two Modern Broilers and Their Jungle Fowl Ancestor.

Front Genet 2019 21;10:480. Epub 2019 May 21.

Department of Poultry Science, University of Arkansas, Fayetteville, AR, United States.

The objective of this study was to evaluate the of intestinal permeability and liver bacterial translocation (BT) across a modern commercial broiler, a commercial broiler of 1995 genetics, and an unselected Jungle Fowl line. Modern 2015 (MB2015) broiler chicken, random bred line initiated from 1995 (RB1995), and the Giant Jungle fowl (JF). Chickens were randomly allocated to four different dietary treatments. Dietary treatments were (1) a control corn-based diet throughout the trial [corn-corn (C-C)]; (2) an early phase malnutrition diet where chicks received a rye-based diet for 10 days, and then switched to the control diet [rye-corn (R-C)]; (3) a malnutrition rye-diet that was fed throughout the trial [rye-rye (R-R)]; and (4) a late phase malnutrition diet where chicks received the control diet for 10 days, and then switched to the rye diet for the last phase [corn-rye (C-R)]. Paracellular permeability was evaluated using fluorescein isothiocyanate dextran (FITC-D). Liver BT was also evaluated. MB2015 and RB1995 consuming the rye-based diet showed increase serum levels of FITC-D when compared to the corn-fed chickens ( < 0.05). Overall, MB2015 appeared to have higher enteric permeability than the JF. To our knowledge, this would be the first paper to evaluate the effect of compensatory growth on intestinal permeability and liver BT. Further studies to evaluate microbiome and inflammatory markers in these chicken models are currently being evaluated.
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http://dx.doi.org/10.3389/fgene.2019.00480DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6536657PMC
May 2019

Feather HSP70: a novel non-invasive molecular marker for monitoring stress induced by heat exposure in broilers.

Poult Sci 2019 Sep;98(9):3400-3404

Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, Arkansas 72701.

Poultry well-being and economic burden due to heat stress (HS) are of great importance to the poultry industry. Efficient design and effective evaluation of any strategies to alleviate the adverse effects of HS on poultry production require an accurate measurement of stress. However, current methods for monitoring stress in poultry are less than ideal, as they are invasive or subjective, and therefore variable. Here, we demonstrated that HSP70, the well-established intracellular stress chaperone, is expressed in chicken feather and is responsive to HS, and could therefore be used as a stress marker. Growing feathers were collected from the same bird (Cobb500, n = 9) in the morning (barn temperature 24°C) and afternoon (barn temperature 29.5°C) in the summer (2016 June 21) in Arkansas, USA. In the afternoon, the birds were panting and their core body temperatures were significantly higher compared to the morning time, as illustrated by iButton data thermo-loggers. Concomitantly, blood HSP70 mRNA and feather HSP70 expression (mRNA and protein) were significantly increased in the afternoon compared to the morning time. Similarly, HSP70 protein expression in the duodenum was also significantly higher in the afternoon compared to the morning period. Together, these finding identify feather HSP70 as a novel non-invasive molecular signature that mirrors the intracellular and systemic stress, which can be useful to monitor well-being of chickens and other avian species under different challenges.
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http://dx.doi.org/10.3382/ps/pez120DOI Listing
September 2019

Enrichment of Autophagy and Proteosome Pathways in Breast Muscle of Feed Efficient Pedigree Male Broilers.

Front Physiol 2018 26;9:1342. Epub 2018 Oct 26.

Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR, United States.

Feed efficiency (FE) is an important genetic trait in poultry and livestock. Autophagy (self-eating) and proteosomes are cellular processes that remove damaged cell components (e.g., proteins, organelles). As evidence of extensive protein oxidation was observed in Pedigree Male (PedM) broilers exhibiting a low FE (LFE) phenotype compared to a high FE (HFE) phenotype, the main goal of this study was to assess gene and protein expression of the autophagy and proteosome pathways in breast muscle obtained in PedM broilers exhibiting HFE and LFE phenotypes. Feed efficiency was calculated as weight gain divided by feed intake gain in individual PedM broilers that were measured between 6 and 7 weeks of age. Targeted gene expression was conducted on breast muscle using quantitative real-time polymerase chain reaction (qPCR) to determine mRNA expression of genes associated with the autophagy pathway; AMP-activated protein kinase alpha 1 (AMPKα1), mammalian target of rapamycin (mTOR), Beclin 1, and autophagy genes (Atg) 3, Atg7, and Atg16L1. Binomial distribution analysis was conducted on transcriptomic and data obtained by RNAseq and shotgun proteomics, respectively on the same set of tissues for genes associated with autophagy, vacuole formation, and proteosome expression. Greater efficiency was attained in the HFE PedM broilers by greater weight gain on the same amount of feed consumed resulting in FEs of 0.65 ± 0.01 and 0.46 ± 0.01 in the HFE and LFE phenotypes, respectively. Targeted mRNA expression analysis revealed significant ( < 0.05) elevations in AMPKa1, mTOR, Atg16L1, and Atg7 and a marginal ( = 0.07) elevation in Beclin1. Binomial distribution analysis transcriptomic and proteomic data revealed significant skews favoring autophagy-, vacuole-, and proteosome-related genes in the HFE phenotype. These results indicate that the autophagy and proteosome expression is enhanced in the HFE compared to the LFE pedigree male broiler phenotype suggesting that protein and organelle quality control may be enhanced in high feed efficiency.
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http://dx.doi.org/10.3389/fphys.2018.01342DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6213487PMC
October 2018