Perssondowd6968

Phytophthora sojae is a widely distributed, destructive oomycete plant pathogen that has been developed as a model for oomycete biology. Given the important but limited reports on the comparison of the sexual and asexual stages in Phytophthora, we performed a large-scale quantitative proteomics study on two key asexual life stages of P. sojae-the mycelium and cyst-as well as on the oospore, which is a key sexual stage. Over 29,631 peptides from 4688 proteins were analyzed. Briefly, 445 proteins, 624 proteins, and 579 proteins were defined as differentially quantified proteins in cyst vs mycelium, oospore vs cyst, and oospore vs mycelium comparisons, respectively (|log2 fold change| > 1 and P  less then  .05). Compared to the mycelium and oospore, fatty acid and nitrogen metabolism were specifically induced in cysts. In oospores, the up-regulated proteins focused on RNA transport and protein processing in endoplasmic reticulum, indicating translation, folding, and the secretion of core cellular or stage-specific proteins active in oospores, which might be used for oospore germination. The data presented expand our knowledge of pathways specifically linked to asexual and sexual stages of this pathogen. BIOLOGICAL SIGNIFICANCE The sexual spores (oospores) in oomycetes have thick cell walls and can survive in the soil for years, thus providing a primary source and allowing the reinfection of their host plant in subsequent growing seasons. However, the proteomic study on oospores remains very limited as they are generally considered to be dormant. In the present study, we successfully isolated oospores, and performed a large-scale comparative quantitative proteomics study on this key sexual stage and two representative asexual stages of P. sojae. The results provide an improved understanding of P. sojae biology and suggest potential metabolic targets for disease control at the three different developmental stages in oomycetes. V.Revolutions in natural and exact sciences started at the dawn of last century have led to the explosion of theoretical, experimental, and computational approaches to determine structures of molecules, complexes, as well as their rich conformational dynamics. Since different experimental methods produce information that is attributed to specific time and length scales, corresponding computational methods have to be tailored to these scales and experiments. These methods can be then combined and integrated in scales, hence producing a fuller picture of molecular structure and motion from the "puzzle pieces" offered by various experiments. Here, we describe a number of computational approaches to utilize experimental data to glance into structure of proteins and understand their dynamics. We will also discuss the limitations and the resolution of the constraints-based modeling approaches. SIGNIFICANCE Experimentally-driven computational structure modeling and determination is a rapidly evolving alternative to traditional approaches for molecular structure determination. These new hybrid experimental-computational approaches are proving to be a powerful microscope to glance into the structural features of intrinsically or partially disordered proteins, dynamics of molecules and complexes. In this review, we describe various approaches in the field of experimentally-driven computational structure modeling. OBJECTIVE Our goal was to identify aspects of residency applications predictive of subsequent performance during pediatric internship. METHODS We conducted a retrospective cohort study of graduates of U.S. medical schools who began pediatric internship in a large pediatric residency program in the summers of 2013 through 2017. GSK-3 inhibition The primary outcome was the weighted average of subjects' ACGME pediatric milestones scores at the end of pediatric internship. To determine factors independently associated with performance, we conducted multivariate linear mixed-effects models controlling for match year and Milestone grading committee as random effects and the following application factors as fixed effects letter of recommendation strength, clerkship grades, medical school reputation, master's or PhD degrees, gender, USMLE Step 1 score, Alpha Omega Alpha membership, private medical school, and interview score. RESULTS Our study population included 195 interns. In multivariate analyses, the aspects of applications significantly associated with composite Milestone scores at the end of internship were LOR strength (estimate 0.09, 95% confidence intervals 0.04, 0.15), numbers of clerkship honors (est. 0.05, 95% CI 0.01-0.09), medical school ranking (est. 0.04, 95% CI 0.08-0.01), having a master's degree (est. 0.19, 95% CI 0.03-0.36), and not having a PhD (est. 0.14, 95% CI 0.02-0.26). Overall the final model explained 18% of the variance in milestone scoring. CONCLUSION Letter of recommendation strength, clerkship grades, medical school ranking, and having obtained a Master's degree were significantly associated with higher clinical performance during pediatric internship. BACKGROUND 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is effective in skin tumours. Studies have demonstrated that the therapy has anti-tumour immunity in squamous cell carcinoma. Exosomes play an important role in tumour microenvironment (TME) crosstalk. Nevertheless, whether exosomes mediate the ALA-PDT anti-tumour effect is unclear. This study aims to investigate whether exosomes secreted from ALA-PDT-treated squamous carcinoma cells (SCCs) demonstrate an anti-tumour effect by inducing dendritic cell (DCs) maturation. METHOD In this study, we used electron microscopy, nanoparticle tracking analysis and western blotting to identify exosomes. Subsequently, BCA assay and fluorescence staining were used to evaluate the biological activity of exosomes. Exosomes derived from ALA-PDT-treated SCCs were incubated with SCCs, fibroblasts and immature DCs, separately. A CCK-8 kit was used to analyse the cytotoxicity of exosomes to SCCs. ELISA was utilised to analyse IL-6, VEGF, MMP-3, and TGF-β1 secreted from fibroblasts. FACS and ELISA were used to analysed DC phenotypic maturation (CD80, MHC-II) and IL-12 secretion. RESULT Herein we show that exosomes secreted from SCCs after ALA-PDT cannot exert cytotoxicity towards SCCs. However, exosomes derived from ALA-PDT-treated SCCs could induce DCs maturation and IL-12 secretion. Furthermore, exosomes secreted from SCCs after ALA-PDT promote the secretion of TGF-β1 from fibroblast. CONCLUSION In conclusion, we found that exosomes derived from ALA-PDT-treated SCCs have the ability to stimulate DC maturation and fibroblast secretion of TGF-β1, which results in the elevation of anti-tumour immunity. These findings provide a new promising strategy of anti-tumour immune response for ALA-PDT in treating SCCs. V.