ABSTRACT
Immunostimulating activities of yeast ß(1 â 3)-D-Glucan (ß-Glucan) mainly depended on its structures. However, due to the tight triple helix structure of ß-Glucan, its immunostimulating activity is greatly weakened. Therefore, in order to partially unwind the tight triple helix structure of ß-glucan and improve its solubility in the medium, we modified it by amination in this study (A-Glu). The results showed that A-Glu could stimulate Raw264.7 macrophages and significantly enhance its TNF-α, IL-6, and IL-10 cytokine expression levels in vitro. A-Glu could also induce a shift of M0 Raw264.7 toward M1, and M2 toward M1. To expand the application of A-Glu in wound repair, the composite sponge consisting of A-Glu and type I collagen via the formation of a stable polyion complex (PIC) was developed. Moreover, the composite sponge could accelerate wound repair significantly. These results reveal that soluble A-Glu as an immunostimulating agent has potential applications in biomedicine.
Subject(s)
beta-Glucans , beta-Glucans/chemistry , Adjuvants, Immunologic/chemistry , Glucans/chemistry , Macrophages/metabolism , Saccharomyces cerevisiae/chemistry , Collagen/metabolismABSTRACT
Eimeria tenella (E. tenella) is one of the most virulent pathogens of coccidiosis. In apicomplexan parasites, Hsp90 (Heat shock protein 90) is essential for the invasion and survival in host cells. In this study, the effect of diclazuril, an effective benzeneacetonitrile anticoccidial agent, on the expression of Hsp90 in the second-generation merozoites of E. tenella was investigated. We inoculated 8 × 10(4) oocysts/chicken suspended in 1 ml of distilled water, and chickens were challenged with E. tenella oocysts and provided with normal feed as Control group; chickens challenged with E. tenella oocysts and provided with 1mg/kg diclazuril in feed from 96 h to 120 h after inoculation as treatment group. Then the second-generation merozoites were obtained after 120 h from the infected caeca. Our results showed that the transcription level of mzHsp90 was reduced by 29.7% in the diclazuril treatment group, accompanied by reduced level of mzHsp90 protein in second-generation merozoites prepared from infected chickens. We also found that the subcellular localization of mzHsp90 was more dispersed in these merozoites. Moreover, we demonstrated that the effects of diclazuril on mzHsp90 expression were direct by in vitro experiments. Taken together, our data provide insights into the molecular mechanisms of diclazuril in the chemotherapy of E. tenella, and suggest that mzHsp90 represents a promising target for the intervention with E. tenella infection.
Subject(s)
Eimeria tenella/drug effects , Eimeria tenella/metabolism , Gene Expression Regulation/drug effects , HSP90 Heat-Shock Proteins/metabolism , Nitriles/pharmacology , Triazines/pharmacology , Animals , Chickens , Coccidiosis/parasitology , Coccidiosis/veterinary , Coccidiostats/pharmacology , HSP90 Heat-Shock Proteins/genetics , Male , Merozoites/drug effects , Merozoites/metabolismABSTRACT
A sensitive liquid chromatography-tandem mass spectrometry method for the simultaneous determination of triclabendazole, its main metabolites (triclabendazole sulphone and triclabendazole sulphoxide) and a marker residue (ketotriclabendazole) in bovine and goat muscle, liver, and kidney samples is developed and validated. Analyte extraction from samples is effectively performed using liquid-liquid extraction by acetonitrile. Chromatographic separation is performed on a C18 reversed-phase column with gradient elution. The analytes are detected by tandem quadrupole mass spectrometry after positive electrospray ionization by multiple reaction monitoring. The limits of detection for analytes are found to be 0.25-2.5 µg/kg in muscle tissues and 1-10 µg/kg in liver and kidney tissues, respectively. The recoveries of edible bovine and goat tissues range from 84.9% to 109.5% when spiked at different levels with analytes, with relative standard deviations generally below 12.8%.
Subject(s)
Anthelmintics/analysis , Anthelmintics/metabolism , Benzimidazoles/analysis , Benzimidazoles/metabolism , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Animals , Cattle , Goats , Kidney/chemistry , Kidney/metabolism , Liver/chemistry , Liver/metabolism , Muscles/chemistry , Muscles/metabolism , TriclabendazoleABSTRACT
A rapid, sensitive and specific liquid chromatography-electrospray ionization (ESI) tandem mass spectrometry (LC-MS-MS) method has been developed for the identification of nitazoxanide metabolites in goat plasma and urine. The purified samples was separated using an XTerra MS C8 column with the mobile phase consisted of acetonitrile and 10-mM ammonium acetate buffer (pH 2.5) followed a linear gradient elution, and detected by MS-MS. Identification and structural elucidation of the metabolites were performed by comparing their retention-times, full scan, product ion scan, precursor ion scan and neutral loss scan MS-MS spectra with those of the parent drug or other available standard. Four metabolites (tizoxanide, tizoxanide glucuronide, tizoxanide sulfate and hydroxylated tizoxanide sulfate) were found and identified in goat after single oral administration of 200mg/kg dose of nitazoxanide. In addition, the possible metabolic pathway was proposed for the first time. The results proved that the established method was simple, reliable and sensitive, revealing that it could be used to rapid screen and identify the structures of active metabolites responsible for pharmacological effects of nitazoxanide and to better understand its in vivo metabolism.
