Effects of growth factors on equine synovial fibroblasts seeded on synthetic scaffolds for avascular meniscal tissue engineering

Across species, the avascular portion of the knee meniscus cannot heal spontaneously if severely injured. The most common treatment is meniscectomy which results in osteoarthritis. The objective of this study was to assess the fibrochondrogenic potential of equine fibroblast-like synoviocytes (FLS) seeded on scaffolds under the influence of growth factors in vitro to determine the potential of developing a novel cell-based repair strategy. Cultured FLS were seeded onto synthetic scaffolds in a rotating bioreactor under the influence of three growth factor regimens: none, basic fibroblast growth factor (bFGF) alone, and bFGF plus transforming growth factor (TGF-β₁) and insulin-like growth factor (IGF-1). Constructs were analyzed for mRNA expression and production of fibrochondroid extracellular matrix constituents. Type II collagen and aggrecan mRNA were significantly higher in growth factor-treated groups (p < 0.05). Despite sub-optimal extracellular matrix production, FLS can exhibit fibrochondral characteristics and may have potential for cell-based tissue engineering for avascular meniscal regeneration.     

In Vitro Evaluation of the Antibiotic Activity of Combinations of Ophthalmic Drugs Against Common Equine Ocular Pathogens

A subpalpebral lavage catheter is commonly used to administer medications in the treatment of infectious keratitis in horses. Drugs may mix in the subpalpebral lavage catheter, potentially affecting antimicrobial efficacy. The objective of this study was to determine the in vitro antimicrobial efficacy of combinations of commonly used ophthalmic preparations. An agar gel diffusion bioassay was used to determine the antibiotic activity of a number of antimicrobial, antifungal, mydriatic, and antiproteinase drug combinations. Drug mixtures were allowed to sit in a syringe at ambient temperatures and room lighting, for 1 (T1) and 6 hours (T2). The antibacterial efficacy was determined by measuring the zone of inhibition around wells containing the drug combinations after overnight incubation at 37°C and comparing them with the zone of inhibition around the antimicrobial alone. Combinations were tested against Pseudomonas aeruginosa and Streptococcus zooepidemicus. There was no significant decrease in the zone of inhibition, compared with controls, for the majority of combinations tested. Significantly smaller zones of inhibition were identified for the following combinations: (1) tobramycin, cefazolin, natamycin, and serum (T1) and (2) tobramycin, cefazolin, natamycin, serum, and atropine (T1) compared with tobramycin alone; (3) ciprofloxacin, cefazolin, natamycin (T1) compared with ciprofloxacin alone; (4) triple antibiotic ophthalmic solution (TAB) and natamycin (T6); (5) TAB, natamycin, and serum (T6); and (6) TAB, natamycin, serum, and atropine (T6) compared with TAB alone. Admixture does not affect in vitro antibacterial efficacy for the majority of ophthalmic preparations, although efficacy of TAB may be decreased compared with the drug alone.     

Scaling-up regional fruit and vegetable distribution: potential for adaptive change in the food system

As demand for locally grown food increases there have been calls to ‘scale-up’ local food production to regionally distribute food and to sell into more mainstream grocery and retail venues where consumers are already shopping. Growing research and practice focusing on how to improve, expand and conceptualize regional distribution systems includes strategies such as value chain development using the Agriculture of the Middle (AOTM) framework. When the Ohio Food Policy Advisory Council asked how they could scale-up the distribution of Ohio fresh fruits and vegetables to Ohioans, we decided to use this practical opportunity to not only provide recommendations to this council, but to simultaneously contribute to the literature on AOTM, value-based and spatially–proximate relationships, and conceptualizations of food system hybridity. We do this while examining an entire sub-sector of the Ohio agricultural economy, namely fruit and vegetables and applying the AOTM framework beyond the farm, namely to distributors and retailers. Through interviews with Ohio retailers and a survey of all fresh fruit and vegetable distributors Ohio we: (1) Describe current distribution systems within the state; (2) Identify firms interested in scaling-up distribution, and; (3) Inform state-level policy efforts by identifying opportunities to better target any state-level policy and program efforts. We demonstrate support for the concept of AOTM applied beyond the farm, for value chain development strategies that can transmit ‘quality’ via spatially proximate supply chains, and support for considering hybrid solutions, such as piggybacking for scaling-up local food systems. This work highlights the role a statewide food policy council can have in facilitating market development and their unique position to provide public sector and institutional support to facilitate meaningful connections in the food system.     

Influence of Stenocarpella maydis Infected Corn on the Composition of Corn Kernel and Its Conversion into Ethanol

Widespread epidemics of Stenocarpella ear rot (formerly Diplodia ear rot) have occurred throughout the central U.S. Corn Belt in recent years, but the influence of S. maydis infected grain on corn ethanol production is unknown. In this study, S. maydis infected ears of variety Heritage 4646 were hand‐harvested in 2010 from a production field in central Illinois and segregated into one of five levels of ear rot severity based upon visual symptoms. The concentration of ergosterol, a sterol produced by fungi but not plants, was observed to increase with the severity of ear rot (127–306.5 μg/g), and none was detected in the control corn. Corn test weight declined with progression of the disease and was 42.6% lower for the most severely rotted grain from ears infected early in their development. Accompanying changes in composition were also apparent. Crude fat and oil contents decreased (from 4.7 to 1.5%) and fiber increased (from 6.6 to 9.6%), but starch content remained largely invariant. Oil composition also varied among the infected samples. Control and infected corn samples were subjected to ethanol fermentation with a laboratory‐scale corn dry‐grind ethanol process. Ethanol yields for control and infected samples were similar on an equivalent weight basis (2.77–2.85 gal/bu). In comparison with the control, S. maydis infection altered the distillers dried grains with solubles (DDGS) properties, wherein the crude protein was significantly higher and oil significantly reduced, and ash, fiber, and yield per ton were not significantly different. Based upon these results, we conclude that Stenocarpella ear rot has the potential to affect DDGS composition but not ethanol yield on an equivalent weight basis.     

Induction, regulation and physiological role of IL-17 secreting helper T-cells isolated from PBMC, thymus, and lung lymphocytes of young pigs

Interleukin-17 (IL-17) producing cells, referred to as Th17, have recently emerged as a third subset of the T helper (Th) cell family. Studies in mice have demonstrated that Th17 cells and their associated cytokines are involved in several autoimmune diseases and host defense against infection. Murine Th17 cells differentiate from naïve CD4⁺ T-cells in the presence of TGFβ and IL-6, however, there are contradicting reports as to the role of TGFβ in the differentiation of human Th17 cells and very little is known about these cells in other animals. We report here the presence of IL-17 secreting lymphocytes in the lung and peripheral blood of pigs. The cDNA of porcine IL-17 gene was cloned and sequenced from activated lung lymphocytes and PBMC from piglets. A 17 kDa recombinant protein was expressed and purified both under denaturing and native conditions from E. coli BL21 cells. Furthermore, we demonstrate that TGFβ in the presence of IL-6 and/or IL-1β induces in vitro differentiation of Th17 cells from naïve porcine CD4⁺ thymocytes.     

FIV diversity: FIV Ple subtype composition may influence disease outcome in African lions

Feline immunodeficiency virus (FIV) infects domestic cats and at least 20 additional species of non-domestic felids throughout the world. Strains specific to domestic cat (FIV(Fca)) produce AIDS-like disease progression, sequelae and pathology providing an informative model for HIV infection in humans. Less is known about the immunological and pathological influence of FIV in other felid species although multiple distinct strains of FIV circulate in natural populations. As in HIV-1 and HIV-2, multiple diverse cross-species infections may have occurred. In the Serengeti National Park, Tanzania, three divergent subtypes of lion FIV (FIV(Ple)) are endemic, whereby 100% of adult lions are infected with one or more of these strains. Herein, the relative distribution of these subtypes in the population are surveyed and, combined with observed differences in lion mortality due to secondary infections based on FIV(Ple) subtypes, the data suggest that FIV(Ple) subtypes may have different patterns of pathogenicity and transmissibility among wild lion populations.     

Pharmacokinetics and clinical effects of pirfenidone administered intravenously in horses

OBJECTIVE: To characterize the plasma pharmacokinetics and clinical effects of pirfenidone administered IV in healthy horses. ANIMALS: 6 adult horses. PROCEDURES: A 15 mg/kg dose of pirfenidone was administered IV over 5 minutes. Physical variables were recorded and blood samples collected prior to infusion; 2.5 minutes after beginning infusion; at the end of infusion; and at 3, 6, 9, 12, 15, 20, 25, 30, 40, 50, 60, 75, and 90 minutes and 2, 2.5, 3, 4, 6, 8, 12, and 24 hours after completion of infusion. Plasma concentrations of pirfenidone and its metabolites were determined. RESULTS: Mild clinical effects, including tachycardia and muscle fasciculations, were observed during drug administration but stopped at the end of the infusion. Pirfenidone and 2 metabolites, hydroxypirfenidone and carboxypirfenidone, were detected by the end of the 5-minute infusion. Mean peak plasma concentration of pirfenidone was 182.5 micromol/L, detected at the end of the infusion. Mean peak plasma concentrations of hydroxypirfenidone and carboxypirfenidone were 1.07 and 3.4 micromol/L, respectively, at 40 minutes after infusion. No parent drug or metabolites were detected at 24 hours. Distribution of pirfenidone best fit a 2-compartment model, and the drug had mean +/- SEM elimination half-life of 86.0 +/- 4.7 minutes, mean body clearance of 6.54 +/- 0.45 mL/kg/min, and apparent volume of distribution at steady state of 0.791 +/- 0.056 L/kg. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous administration of pirfenidone was tolerated with transient adverse affects during infusion, and drug clearance was rapid.