Project description:We investigated the biological effects of ZEA exposure on donkey granulosa cells by using RNA-seq analysis. ZEA at 10 and 30 μM were administered to granulosa cells within 72 hours of in vitro culture. ZEA at 10 μM significantly altered the tumorigenesis associated genes in donkey granulosa cells. Exposure to 10 and 30 μM ZEA treatment significantly reduced mRNA expression of PTEN, TGFβ, ATM, and CDK2 genes, particularly, the ZEA treatment significantly increased the expression of PI3K and AKT genes. Furthermore, immunofluorescence, RT-qPCR, and Western blot analysis verified the gene expression of ZEA-exposed granulosa cells. Collectively, these results demonstrated the deleterious effect of ZEA exposure on the induction of ovarian cancer related genes via the PTEN/PI3K/AKT signaling pathway in donkey granulosa cells in vitro.
Project description:In this study, 3,869 donkey skeletal muscle lncRNAs were identified using RNA-Seq along with a stringent screening procedure in the longissimus dorsi (LD) and gluteal (G) muscles. These lncRNAs share many characteristics with other mammalian lncRNAs, such as shorter open reading frames (ORFs) and lower expression levels than mRNAs. Furthermore, in pairwise comparisons between libraries of the same stage for two genetic types of male Dezhou donkey, 73 differentially expressed lncRNAs were common to all muscle tissues.
Project description:We examined the growth curve, cell cycle, apoptosis and glycolysis of donkey, horse and mule adult fibroblasts (DAFs, HAFs and MAFs), which indicated there are differences in cell proliferation and metabolism. We also derived mule, donkey and horse iPSCs from their respective adult fibroblasts by piggyBac transposition, and we found the induced reprogramming efficiency of mule iPSCs was significantly higher than donkey and horse iPSCs (78.3% vs 58.2% vs 47.9%). miPSCs, diPSCs and hiPSCs all expressed high levels of key endogenous pluripotency genes such as Oct4, Sox2 and Nanog, propagated robustly in single cell passaging and miPSCs were found to proliferated significantly faster than diPSCs and hiPSCs. Furthermore, miPSCs/MAFs clustered closer to diPSCs/DAFs than to hiPSCs/HAFs by RNA-seq. The establishment of miPSCs provide unique experimental materials for further investigation of understanding the “heterosis” and reproductive isolation during speciation.
Project description:The skin barrier is vital for protection against environmental threats including insults caused by skin-resident microbes. Dysregulation of this barrier is a hallmark of atopic dermatitis (AD) and ichthyosis, with variable consequences for host immune control of colonizing commensals and opportunistic pathogens. While Malassezia is the most abundant commensal fungus of the skin, little is known about the host control of this fungus in inflammatory skin diseases. Here we show that in barrier-impaired skin, Malassezia acquires enhanced fitness and overt growth properties. By using four distinct and complementary murine models of atopic dermatitis and ichthyosis we provide evidence that structural and metabolic changes in the dysfunctional epidermal barrier environment provide increased accessibility and an altered lipid profile, to which the lipid-dependent yeast adapts for enhanced nutrient assimilation. These findings reveal fundamental insights into the implication of the mycobiota in the pathogenesis of common skin barrier disorders.
Project description:Trichophyton rubrum is a pathogenic fungus infecting human skin, hairs and nails. These substrates are low in most nutrients required for fungal growth and consequently are colonized only by very few fungal species. Especially, concentration of trace elements is low and a limiting factor for fungal growth. T. rubrum is a highly specialist fungus and adapted to this environment. By in-vitro experiments, we analyzed the influence of trace-elements on mRNA expression. We measured gene expression by RNAseq of two T. rubrum strains, STRB008 and STRB012, in three different cultivation condition, each in 6 replications. Keratin medium, without sugar supply, was used as basic medium. In the second condition, we added trace-elements to the keratin medium. In the third condition, we added glucose. We point to the evolutionary adaption of the fungus to the human skin. T. rubrum has a sophisticated system for the digestion and utilization of human skin protein and a relative low demand for trace-elements.
Project description:A comprehensive glycosylation profile of donkey lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. The results obtained allowed the identification of 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. Altogether, the N-glycan structures determined revealed that in donkey milk lactoferrin most of the N-glycans identified are neutral complex/hybrid. Actually, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. In addition, 2 high mannose N-glycans, 4 sialylated fucosylated complex/hybrid N-glycans and 6 sialylated non-fucosylatedN-glycans, one of which containing N-glycolylneuramin acid (Neu5Gc), were found. A comparison of the glycosylation profile of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported.
Project description:Background: milk is considered an important source of bioactive peptides, which can be produced by endogenous or starter bacteria, such as lactic acid bacteria, that are considered effective and safe producers of food-grade bioactive peptides. Among the various types of milk, donkey milk has been gaining more and more attention for its nutraceutical properties. Methods: Lactobacillus rhamnosus 17D10 and Lactococcus lactis subsp. cremoris 40FEL3 were selected for their ability to produce peptides from donkey milk. The endogenous peptides and those obtained after bacterial fermentation were assayed for their antioxidant, antibacterial and antiviral activities. The peptide mixtures were characterized by means of LC-MS/MS, and then analyzed in silico using the Milk Bioactive Peptide DataBase. Results: the peptides produced by the two selected bacteria enhanced the antioxidant activity and reduced E. coli growth. Only the peptides produced by L. rhamnosus 17D10 were able to reduce S. aureus growth. All the peptide mixtures were able to inhibit the replication of HSV-1 by more than 50%. Seventeen peptides were found to have 60% sequence similarity with already known bioactive peptides. Conclusion: a lactic acid bacterium fermentation process is able to enhance the value of donkey milk through bioactivities that are important for human health.