Evaluation of porcine epidemic diarrhea virus transmission and the immune response in growing pigs

Clinical disease associated with porcine epidemic diarrhea virus (PEDV) infection in naïve pigs is well chronicled; however, information on endemic PEDV infection is limited. To characterize chronic PEDV infection, the duration of infectious virus shedding and development of protective immunity was determined. On Day 0 (D0), a growing pig was challenged with PEDV and 13 contacts were commingled. On D7, 9 contact pigs (principal virus group (PG)), were selected, moved to a separate room and commingled with one sentinel pig (S1). This process was repeated weekly with S2, S3 and S4. The PG was PEDV-positive by PCR from D3-11, with some pigs intermittently positive to D42. Pigs S1 and S2 were PEDV-positive within 24 hours of commingling. Antibodies were detected in all PG by D21 and by 7 days post-contact in S1 and S2. Pigs S3 and S4 were PCR and antibody negative following commingling. To evaluate protective immunity, 5 naïve pigs (N) and the PG were challenged (N/C, PG/C) with homologous virus on D49. All N/C pigs were PEDV PCR-positive by D52 with detection out to D62 in 3/5 N/C pigs. All PG/C pigs were PEDV PCR-negative post-challenge. By D63, all N/C seroconverted. Although PEDV RNA was demonstrated in pigs after primary infection until D42, infectious PEDV capable of horizontal transmission to naïve pigs was only shed 14–16 days after infection to age-matched pigs. Homologous re-challenge 49 days post initial PEDV exposure did not result in re-infection of the pigs. This demonstrates potential for an effective PEDV vaccine.     

Identification of Plakortide E from the Caribbean Sponge Plakortis halichondroides as a Trypanocidal Protease Inhibitor using Bioactivity-Guided Fractionation

In this paper, we report new protease inhibitory activity of plakortide E towards cathepsins and cathepsin-like parasitic proteases. We further report on its anti-parasitic activity against Trypanosoma brucei with an IC₅₀ value of 5 μM and without cytotoxic effects against J774.1 macrophages at 100 μM concentration. Plakortide E was isolated from the sponge Plakortis halichondroides using enzyme assay-guided fractionation and identified by NMR spectroscopy and mass spectrometry. Furthermore, enzyme kinetic studies confirmed plakortide E as a non-competitive, slowly-binding, reversible inhibitor of rhodesain.     

Invertase activity limits grain yield of maize under salt stress

Salt stress reduces grain yield of maize (Zea mays L.) due to poor kernel setting but not due to decreased grain filling. In the present study, it was tested whether acid invertase activity is decreased in developing kernels of maize under salt stress, and if assimilate supply is limited. The relatively salt‐sensitive maize hybrid Pioneer 3906 was compared with the more salt‐resistant hybrid SR 12. Salt stress caused a significant decrease in grain yield which was due to a 50% decrease in kernel number. No source limitation was observed, as the sucrose concentrations in kernels were significantly increased under salt stress for both genotypes. In contrast, glucose and fructose concentrations in kernels were significantly decreased. Salt stress caused a significant inhibition of soluble acid invertase activity to 19% in hydroponics 5 d after pollination (5 DAP) and to 50% in the soil culture experiment (2 DAP). The decrease in enzyme activity was the same for both genotypes. In the soil experiment, the highest soluble acid invertase activity was found 2 DAP with a steep decline until 8 DAP in Pioneer 3906. It is concluded that a decrease in acid invertase activity is a key factor associated with limited kernel setting under salt stress but additional factors may be responsible for genotypic differences.     

Fructokinase,Fructans, Intestinal Permeability,and Metabolic Syndrome: An Equine Connection?

Fructose is a simple sugar present in honey and fruits, but can also exist as a polymer (fructans) in pasture grasses. Mammals are unable to metabolize fructans, but certain gram-positive bacteria contain fructanases and can convert fructans to fructose in the gut. Recent studies suggest that fructose generated from bacteria or directly obtained from the diet can induce both increased intestinal permeability and features of metabolic syndrome, especially the development of insulin resistance. The development of insulin resistance is driven in part by the metabolism of fructose by fructokinase C in the liver, which results in oxidative stress in the hepatocyte. Similarly, the metabolism of fructose in the small bowel by intestinal fructokinase may lead to increased intestinal permeability and endotoxemia. Although speculative, these observations raise the possibility that the mechanism by which fructans induce laminitis could involve intestinal and hepatic fructokinase. Further studies are indicated to determine the role of fructanases, fructose, and fructokinase in equine metabolic syndrome and laminitis.     

Synthesis and Cytotoxicity Evaluation of Spirocyclic Bromotyrosine Clavatadine C Analogs

Marine-originated spirocyclic bromotyrosines are considered as promising scaffolds for new anticancer drugs. In a continuation of our research to develop potent and more selective anticancer compounds, we synthesized a library of 32 spirocyclic clavatadine analogs by replacing the agmatine, i.e., 4-(aminobutyl)guanidine, side chain with different substituents. These compounds were tested for cytotoxicity against skin cancer using the human melanoma cell line (A-375) and normal human skin fibroblast cell line (Hs27). The highest cytotoxicity against the A-375 cell line was observed for dichloro compound 18 (CC₅₀ 0.4 ± 0.3 µM, selectivity index (SI) 2). The variation of selectivity ranged from SI 0.4 to reach 2.4 for the pyridin-2-yl derivative 29 and hydrazide analog of 2-picoline 37. The structure–activity relationships of the compounds in respect to cytotoxicity and selectivity toward cancer cell lines are discussed.     

Bekämpfung von Epilachna vigintioctopunctata (F.) und Spodoptera litura (F.) mit Silikaten

The present work deals with the insecticidal efficacy of different modified natural silica products against the field pests Epilachna vigintioctopunctata (F.) and Spodoptera litura (F.). The substances “Fossil Shield^® 90.0s”, “Advasan^®” as well as different formulations of a silica-derived substance named “AL06” (developed in the section Urban Horticulture, Humboldt-University Berlin) were dusted into gauze-covered cages. In each cage, test insects were placed and mortality measured over time. A 100% mortality rate was archieved two days after treatment of adult E. vigintioctopunctata and S. litura larvae with some silica “AL06”-formulations. In contrast, mortality rate for E. vigintioctopunctata larva was only 30 to 70% after two days of treatment. Mortality rate was dose dependend, even though a linear correlation could not be found. Furthermore, the commercial substance “Advasan^®” was able to control the horticultural pest E. vigintioctopunctata within 48 hours successfully. For future horticultural applications, possible phytotoxic side effects of silica dusts and an application technology resulting in adequate leaf coverage have to be considered.     

Iron Mobilization and Mineralogical Alterations Induced by Iron-Deficient Cucumber Plants (Cucumis sativus L.) in a Calcareous Soil

Dicotyledons cope with ion (Fe) shortage by releasing low-molecular-weight organic compounds into the rhizosphere to mobilize Fe through reduction and complexation mechanisms. The effects induced by these root exudates on soil mineralogy and the connections between Fe mobilization and mineral weathering processes have not been completely clarified. In a batch experiment, we tested two different kinds of organic compounds commonly exuded by Fe-deficient plants, i.e., three organic acids (citrate, malate, and oxalate) and three flavonoids (rutin, quercetin, and genistein), alone or in combination, for their ability to mobilize Fe from a calcareous soil and modify its mineralogy. The effect of root exudates on soil mineralogy was assessed in vivo by cultivating Fe-deficient and Fe-sufficient cucumber plants (Cucumis sativus L.) in a RHIZOtest device. Mineralogical analyses were performed by X-ray powder diffraction. The batch experiment showed that citrate and, particularly, rutin (alone or combined with organic acids or genistein) promoted Fe mobilization from the soil. The combinations of rutin and organic acids modified the soil mineralogy by dissolving the amorphous fractions and promoting the formation of illite. These mineralogical alterations were significantly correlated with the amount of Fe mobilized from the soil. The RHIZOtest experiment revealed a drastic dissolution of amorphous components in the rhizosphere soil of Fe-deficient plants, possibly caused by the intense release of phenolics, amino acids, and organic acids, but without any formation of illite. Both batch and RHIZOtest experiments proved that exudates released by cucumber under Fe deficiency concurred to the rapid modification (on a day-scale) of the mineralogy of a calcareous soil.     

Cationized albumin-biocoatings for the immobilization of lipid vesicles

Tethered lipid membranes or immobilized lipid vesicles are frequently used as biomimetic systems. In this article, the authors presented a suitable method for efficient immobilization of lipid vesicles onto a broad range of surfaces, enabling analysis by quantitative methods even under rigid, mechanical conditions-bare surfaces such as hydrophilic glass surfaces as well as hydrophobic polymer slides or metal surfaces such as gold. The immobilization of vesicles was based on the electrostatic interaction of zwitterionic or negatively charged lipid vesicles with two types of cationic chemically modified bovine serum albumin (cBSA) blood plasma proteins (cBSA-113 and cBSA-147). Quantitative analysis of protein adsorption was performed as the cBSA coatings were characterized by atomic force microscopy, surface zeta potential measurement, fluorescence microscopy, and surface plasmon spectroscopy, revealing a maximal surface coverage 270-280?ng/cm(2) for 0.02 mg/ml cBSA on gold. Small unilamellar vesicles as well as giant unilamellar vesicles (GUVs) were readily immobilized (～15?min) on cBSA coated surfaces. GUVs with 5-10 mol% negatively charged 1,2,-dipalmitoyl-sn-glycero-3-phosphoglycerol remained stable in liquid for at least 5 weeks.