Enhancing antimicrobial activity and reducing cytotoxicity of silver nanoparticles through gelatin nanoparticles - supplementary figures

Aim: To develop a novel stabilizing agent for silver nanoparticles (AgNPs) with the aim of enhancing its antibacterial efficacy against wound associated pathogens while mitigating their cytotoxic effect on human cells. Materials & methods: In this study, monodispersed gelatin nanoparticles were synthesized to stabilize AgNPs. The stability, antibacterial activity and biocompatibility of the gelatin-stabilized AgNPs (Gel-AgNPs) were compared with citrate-stabilized AgNPs (citrate-AgNPs) or silver ions. Results & conclusion: Gelatin-stabilized AgNPs showed significantly better antibacterial activities compared with citrate-stabilized AgNPs against both Gram-positive and Gram-negative bacteria. These Gel-AgNPs showed significantly lower cytotoxicity to human dermal fibroblasts compared with Ag+. These findings provided the first evidence substantiating a novel functionality of gelatin nanoparticles in both stabilizing and enhancing the activity of AgNPs.</p

Dynamic Changes in the Follicular Transcriptome and Promoter DNA Methylation Pattern of Steroidogenic Genes in Chicken Follicles throughout the Ovulation Cycle

<div><p>The molecular mechanisms associated with follicle maturation and ovulation are not well defined in avian species. In this study, we used RNA-seq to study the gene expression profiles of the chicken follicles from different developmental stages (pre-hierarchical, pre-ovulatory and post-ovulatory). Transcriptomic analysis revealed a total of 1,277 and 2,310 genes were differentially expressed when follicles progressed through the pre-hierarchical to hierarchical and pre-ovulatory to post-ovulatory transitions, respectively. The differentially expressed genes (DEG) were involved in signaling pathways such as adherens junction, apoptosis and steroid biosynthesis. We further investigated the transcriptional regulation of follicular steroidogenesis by examining the follicle-specific methylation profiles of Star (steroidogenic acute regulatory protein), Cyp11a1 (cytochrome P450, family 11, subfamily a, polypeptide 1) and Hsd3b (hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1), genes encoding the key enzymes for progesterone synthesis. The varied patterns of DNA methylation in proximal promoters of Star and Cyp11a1but not Hsd3b in different follicles could play a major role in controlling gene expression as well as follicular steroidogenic activity. Finally, the promoter-reporter analysis suggests that TGF-β could be involved in the regulation of Hsd3b expression during ovulation. Together, current data not only provide novel insights into the molecular mechanisms of follicular physiology in chicken follicles, but also present the first evidence of epigenetic regulation of ovarian steroidogenesis in avian species.</p></div

Effect of TGF-β1 on the transcription of Hsd3b gene.

<p><b>(</b>A) Granulosa cells were transfected with Hsd3b promoter reporter constructs containing 5′ serial deletions. Twenty-four hours later, cells were treated with TGF-β1 (5 ng/ml). A renilla luciferase reporter plasmid was used as the internal control to correct for transfection efficiency. (B) qPCR shows Hsd3b gene expression was inhibited by TGF-β1. All the data are presented as the mean ± SEM from at least four independent experiments. The housekeeping gene GAPDH was used for normalization. Student t-test was used to analyze the luciferase activity in TGF-β1 treated cells compared to control. One-way ANOVA followed by Tukey multiple range test was used to analyze Hsd3b gene expression in granulosa cells after different TGF-β1 treatments. * <i>P</i> < 0.05, ** <i>P</i> < 0.01.</p

Methylation patterns of the Star, Cyp11a1 and Hsd3b promoters in chicken follicles.

<p>Site-specific methylation levels of the proximal promoters of Star, Cyp11a1 and Hsd3b from SWF, F1 and POF1 follicles were compared. The Sequenom MassARRAY platform was used for the quantitative methylation analysis. The CpG units locations are as defined in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146028#pone.0146028.s001" target="_blank">S1 Fig</a>. Data are expressed as mean ± SEM. n = 3 animals.</p

Comparisons among 3 tools: CHASM, MutationTastor, and CanDrA.

<p>Comparisons among 3 tools: CHASM, MutationTastor, and CanDrA.</p

Feature optimization for GBM. Plotted are the areas under the curves (AUCs) of the receiver operator characteristics acquired through our incremental feature selection process.

<p>Three sets of AUCs are computed from the 10-fold cross-validation (CV) of the training set GBM.Ex (dotted line) and the independent validation (IV) of 2 test sets, GBM.S1 and GBM.S2 (solid and dashed line). On the x-axis are features that are incrementally selected. The dashed box marks the peaks of the cross-validation AUC, which corresponds to the optimal feature set used for CanDrA.</p

Correlation between mutation score and prevalence.

<p>Twelve algorithms (x-axis) were compared using 4 data sets: (a) GBM mutations in <i>TP53</i>, (b) GBM mutations in <i>PTEN</i>, (c) OVC mutations in <i>TP53</i>, and (d) OVC mutations in <i>KRAS</i>.</p

Feature optimization for OVC. Plotted are the areas under the curves (AUCs) of the receiver operator characteristics acquired through our incremental feature selection process.

<p>Three sets of AUCs are computed from the 10-fold cross-validation (CV) of the training set OVC.Ex (dotted line) and the independent validation (IV) of 2 test sets, OVC.S1 and OVC.S2 (solid and dashed line). On the x-axis are features that are incrementally selected. The dashed box marks the peaks of the cross-validation AUC, which corresponds to the optimal feature set used for CanDrA.</p

Comparison of cancer type specificity between CHASM and CanDrA.

<p>Comparison of cancer type specificity between CHASM and CanDrA.</p

Effect of acid and alkali on activity of Pc-pis-His and Pc-pis against <i>S. aureus</i>.

<p>“—” represents no visible microbe is detected at both blank control group (no peptides) and experimental group.</p