Advancing PEDV Vaccination: Comparison between Inactivated and Flagellin N-Terminus-Adjuvanted Subunit Vaccines.

Vaccinations can serve as an important preventive measure against the porcine epidemic diarrhea (PED) virus that currently threatens the swine industry. This study focuses on the development of a fusion protein vaccine, FliC99-mCOE, which combines the N-terminus of flagellin (FliC99) with a modified core neutralizing epitope (mCOE) of PEDV. In silico immunoinformatic analysis confirmed the construct's non-toxic, non-allergenic, and highly antigenic nature. Molecular docking and molecular dynamics (MD) simulations demonstrated FliC99-mCOE's strong binding to the TLR-5 immunological receptor. Repeated exposure simulations and immunological simulations suggested enhanced cell-mediated immunity. Both FliC99-mCOE and an inactivated PEDV vaccine were produced and tested in mice. The results from cell proliferation, ELISA, and neutralization assays indicated that FliC99-mCOE effectively stimulated cellular immunity and neutralized PEDV. We conclude that the FliC99-mCOE fusion protein may serve as a promising vaccine candidate against PEDV.

The Effectiveness of Sperm Preparation Using Density Mini-Gradient and Single-Layer Centrifugation for Oligospermia Samples.

Background: Sperm preparation is an important step during assisted reproduction, and different assisted reproductive techniques have different sperm quality requirements. For intrauterine insemination (IUI), the total motile sperm count is a predictor of a patient's fertility. Objective: The aim of this study was to compare the sperm recovery rate and DNA fragmentation index (DFI) outcomes following density mini-gradient and single-layer centrifugation in preparation for intrauterine insemination (IUI). Methods: A total of 30 semen samples with concentrations under 15 million cells/ml were obtained, and each sample was divided into 3 aliquots, with each aliquot subjected to 1 of 3 separation methods: mini-gradient, single-layer using a 90% density layer (single 90-layer), and single-layer using a 45% density layer (single 45-layer). Total sperm motility and sperm recovery rates were compared before and after preparation using each method. Results: The sperm concentration obtained using single 45-layer was higher than the other groups (p<0.05), but sperm motility was higher using the mini-gradient and single 90-layer methods higher than the single 45-layer method (p<0.05). The recovered sperm motility rates for the mini-gradient, single 90-layer, and single 45-layer methods were 57.6% ± 20.6%, 62.8% ± 18.5%, and 78.7% ± 12.4%, respectively, indicating a better outcome for the single 45-layer method than for the other methods. Conclusion: All of these methods can be applied to sperm preparation for IUI, and the optimal method can be selected based on initial sperm quality to collect sperm with good motility and DNA integrity to achieve a satisfactory pregnancy rate.

Glycoside constituents from Miliusa sinensis leaves and their anti-inflammatory and acetylcholine protective effects.

Repeated column chromatography resulted in the isolation of two new glycosides, miliusides A-B (1 and 7), along with six known metabolites (2-6, and 8) from the leaves of Miliusa sinensis Finet and Gagnep. The structures of the purified phytochemicals were elucidated by interpreting their spectroscopic data (NMR, HRMS), as well as comparison with the previous literature. The biological evaluation of acetylcholinesterase (AChE) inhibitory effects and anti-inflammatory activity by measuring nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 mouse macrophage cells, were also conducted. Among them, compounds 5 and 7 exhibited significant AChE inhibitory activities (IC50 = 53.36 ± 4.20 and 88.50 ± 8.79 µM, respectively), compared with the positive control (Galanthamine, IC50 = 1.65 ± 0.15 µM). Only the MeOH extract showed suppression effects on NO production in LPS-induced RAW264.7 cells (IC50 = 38.18 ± 3.25 µg/mL) comparable to that of the positive control, l-NMMA (IC50 = 2.21 ± 0.56 µg/mL).

Proliferation and Differentiation of Dopaminergic Neurons from Human Neuroepithelial Stem Cells Obtained from Embryo Reduction Following In Vitro Fertilization.

BACKGROUND: Recent advances in stem cell technologies have rekindled an interest in the use of cell therapies to treat patients with Parkinson's disease. Although the transplantation of dopaminergic mesencephalic human fetal brain tissue has previously been reported in the treatment of patients with Parkinson's disease, this method is limited by the availability of tissue obtained from each human embryo. OBJECTIVE: Our study aimed to isolate, culture, proliferate, and differentiate dopaminergic neurons from human neuroepithelial stem cells obtained from embryo reduction procedures performed in multifetal pregnancies following in vitro fertilization. MATERIALS AND METHODS: A total of 201 human embryos were dissected for isolation and culture of neuroepithelial stem cells for proliferation and differentiation into dopaminergic neurons. All embryos were obtained from embryo reduction procedures performed in multifetal pregnancies after in vitro fertilization treatments. RESULTS: Human neuroepithelial stem cells were isolated and cultured from embryos from 6.0 to 8.0 weeks. Neuroepithelial stem cells were successfully isolated, proliferated, and differentiated into dopaminergic neurons. The cells adhered to the surfaces of cell culture plates after 2 days and could be proliferated and differentiated into neurons within 4 days. Cultured cells expressed the dopaminergic marker tyrosine hydroxylase after 6 days, suggesting that these cells were successfully differentiated into dopaminergic neurons. CONCLUSION: The successful isolation, culture, proliferation, and differentiation of human dopaminergic neurons from embryo reductions performed for multifetal pregnancies after in vitro fertilization suggests that this pathway may serve as a potential source of cell therapy materials for use in the treatment of Parkinson's disease.