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Piplartine, an alkaloid produced by plants in the genus Piper, displays promising anticancer activity. Understanding the gas-phase fragmentation of piplartine by electrospray ionization tandem mass spectrometry can be a useful tool to characterize biotransformed compounds produced by in vitro and in vivo metabolism studies. As part of our efforts to understand natural product fragmentation in electrospray ionization tandem mass spectrometry, the gas-phase fragmentation of piplartine and its two metabolites 3,4-dihydropiplartine and 8,9-dihydropiplartine, produced by the endophytic fungus Penicillium crustosum VR4 biotransformation, were systematically investigated. Proposed fragmentation reactions were supported by ESI-MS/MS data and computational thermochemistry. Cleavage of the C-7 and N-amide bond, followed by the formation of an acylium ion, were characteristic fragmentation reactions of piplartine and its analogs. The production of the acylium ion was followed by three consecutive and competitive reactions that involved methyl and methoxyl radical eliminations and neutral CO elimination, followed by the formation of a four-member ring with a stabilized tertiary carbocation. The absence of a double bond between carbons C-8 and C-9 in 8,9-dihydropiplartine destabilized the acylium ion and resulted in a fragmentation pathway not observed for piplartine and 3,4-dihydropiplartine. These results contribute to the further understanding of alkaloid gas-phase fragmentation and the future identification of piplartine metabolites and analogs using tandem mass spectrometry techniques. Copyright © 2017 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The most accepted hypothesis for Diseases Hemorragic Fever/Dengue Shock Syndrome (DHF/ DSS) pathogenesis is a phenomenon known as Antibody Dependent Enhancement (ADE). MAbs are a useful and sensitive tool for ADE studies. The majority of monoclonal antibodies, which have been used to report ADE activity, recognise epitopes on surface glycoproteins; two of these proteins have been E and prM. A mouse monoclonal antibody specific (MAb 4B6) to a recombinant envelope protein (E) from dengue virus type 4 expressed in Pichia pastoris was studied for ADE of infection in P388D1, mouse macrophage like cells. This study was performed using a viral multiplicity of m=0,1, MAb dilutions to 10-3 to 10-6 and dengue 2 and dengue 4 strains. Our 4B6 monoclonal antibody neutralized homologous virus but it did not enhance it The neutralization and non amplifying characteristic of 4B6 MAb are similar in pattern to others. MAb mimic peptides, chimeric and fusion proteins containing the epitopes for neutralizing and no amplificating Mab could be used as a good subunit candidates for a future dengue vaccines.The fact that this Mab did not revealed the presence of enhancement epitopes in E Dengue 4 recombinant protein suggest that one as an attractive antigen in immunoprotection studies. [ABSTRACT FROM AUTHOR]

Background: Dengue is the most important human viral disease transmitted by arthropod vectors. The availability of random peptide libraries (RPL) displayed on phage has provided a powerful tool for selecting sequences that mimic epitopes from microorganisms that are useful for diagnostic and vaccine development purposes. In this paper, we describe peptides that resemble the antigenic structure of B-cell epitopes of dengue virus identified from a phage-peptide library using human sera containing polyclonal antibodies against dengue virus. Materials and Methods: Eighteen phage clones were isolated from the phage-display peptide library, J404, by affinity selection using human antisera against dengue virus type 3. These clones were tested for reactivity by ELISA with a panel of hyperimmune ascitic fluids (HAFs) containing antibodies either against all four dengue serotypes, West Nile virus (WNV) or Eastern equine encephalitis virus (EEEV) with control ascitic fluid (NAF) used as a negative control. Results: Eight clones were recognized by HAFs against the four dengue serotypes, of which four significantly inhibited binding of anti-dengue antibodies to the virus. Two peptides with similar sequences to regions of NS3 and NS4B non-structural dengue virus proteins were identified. Conclusion: Our results suggest that these peptides could be used for the development of diagnostic tools for the detection of dengue virus infection and for a potential vaccine against this pathogen. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effects of dietary fiber source and direct-fed microbial supplementation on lactation performance and feeding behavior of high-producing dairy cows. Sixty-four multiparous Holstein cows (3.5 ± 1.6 lactations; 76 ± 22 DIM and 735 ± 67 kg of BW at covariate period initiation) and 32 gate feeders were enrolled in a study with a completely randomized design and a 2 × 2 factorial arrangement. Cows and gate feeders were randomly assigned to treatments (16 cows and 8 gate feeds per treatment). Cows were allowed 1 week to acclimate to feeding gates followed by a 2-week covariate period. During the acclimation and covariate periods, all cows were fed the same diet to meet or exceed the nutrient requirements. Following the covariate period, cows were enrolled in a 8-week treatment period during which cows were randomly assigned to 1 of 4 treatments consisting of forage inclusion in the diet, either 45.8% (LF) or 56.7% (HF) of DM, and the supplementation of 75 mL/hd/d of a direct-fed microbial (DFM) containing herbal extracts (mallow, mint, and sage), L. plantarum, L. buchneri, Saccharomyces cerevisiae, and sugar cane molasses (Valibiom Mix, Valibiotics, Traiskirchen, Austria) or without supplementation (CON). The average covariate period value of each variable was used as a covariate. Three-way interactions were observed for DMI and feed efficiency. Dry matter intake was 2 to 3 kg greater for LF-DFM than HF-CON and HF-DFM during wk 2, 3, 5, and 8 of the treatment period. Milk production was 2.1 kg/d greater for LF than HF diets. Both milk fat and MUN concentrations were greater for HF than LF diets. Conversely, milk protein concentration was lower for HF than LF diets. The respiration rate measured in the morning was lower with DFM supplementation than CON. Rectal temperature measured in the morning and averaged for the day were greater for LF than HF diets. Under the conditions of the present study, feeding high-forage diets may be an alternative for producers to reduce feeding costs depending on the price of purchased feeds. However, non-forage fiber sources (i.e., soy hulls) must be considered when producers are challenged by either forage shortages or forage with a lower nutritive value. Additionally, DFM supplementation reduced respiration rate in the morning and affected meal behavior of lactating cows., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)