IL-4 up-regulates epidermal chemotactic, angiogenic, and pro-inflammatory genes and down-regulates antimicrobial genes in vivo and in vitro: Relevant in the pathogenesis of atopic dermatitis

Atopic dermatitis (AD) is a common chronic inflammatory skin disease. Although the pathogenesis of AD is not fully understood, we and others have shown that IL-4 plays a key role. In this study we aimed to identify keratinocyte genes regulated by IL-4 that may play important roles in the pathophysiology of AD. HaCat cells were treated with IL-4 at various concentrations for 24h, and PCR gene array on inflammation/autoimmunity was performed three times for analysis of differential gene expression. Of all the 370 genes examined, 32 and 53 genes are up- and down-regulated, respectively. Specifically related to AD, chemokines CCL3L1, CCL8, CCL24, CCL25, CCL26, CXCL6 and CXCL16 are up-regulated by IL-4. Pro-inflammatory factors, such as IL-19, IL-20, IL-1α, IL-12Rβ2, IL-25, IL-31RA, OSMR and nitric oxide synthase 2, are also up-regulated. In addition, IL-4 up-regulates VEGFA, a pro-angiogenic factor. In contrast, antimicrobial peptides (AMPs) or factors involved in APM production, such as IFN-κ, S100s, Toll-like receptors, and several chemokines are down-regulated. Similarly IL-4 also down-regulates TNF-α, lymphotoxin-β, an IgE suppressor, TNFSF18, a T-cells function regulator, and the glucocorticoid receptor. On the in vivo level, real-time RT-PCR on the selected genes confirmed that IL-4 up-regulates chemokines, proinflammatory cytokines while it suppresses AMP production related genes in the skin obtained from IL-4 Tg mice. Detailed examination of these genes will delineate their specific roles in chemotaxis, inflammation, angiogenesis and AMP production, all of which may contribute to the development and progression of AD

Table_1_Dietary change influences the composition of the fecal microbiota in two rescued wild raccoon dogs (Nyctereutes procyonoides).DOCX

The gut microbiota of wild animals, influenced by various factors including diet, nutrition, gender, and age, plays a critical role in their health and disease status. This study focuses on raccoon dogs (Nyctereutes procyonoides), a commonly found wild animal, and its gut microbiota composition in response to dietary shifts. The study aimed to compare the fecal bacterial communities and diversity of rescued raccoon dogs fed three different diet types (fish and amphibians, mixed protein with maize, and solely maize) using high-throughput sequencing. Results indicated that the dietary composition significantly influenced the gut microbiota, with notable differences in the abundance of several key phyla and genera. The study identified Firmicutes as the dominant phylum in all diet groups, with notable variations in the relative abundances of Bacteroidota, Proteobacteria, and Verrucomicrobiota. Notably, the group solely fed maize exhibited a significant increase in Proteobacteria, potentially linked to dietary fiber and lignin degradation. The genus-level analysis highlighted significant differences, with Lactobacillus and Bifidobacterium responding to dietary shifts. The genus Akkermansia in Verrucomicrobiota can be identified as a marker for assessing the health of the gut and deserves further investigation. Gender-specific differences in the gut microbiota were observed, highlighting the influence of individual variation. Furthermore, the analysis of bacterial functions suggested a connection between diet and host metabolism, emphasizing the need for further research to understand the complex mechanisms underlying the relationship between dietary composition and gut microbiota in wild animals. These findings provide crucial insights into conservation and rescue efforts for wild animals.</p

Image_1_Dietary change influences the composition of the fecal microbiota in two rescued wild raccoon dogs (Nyctereutes procyonoides).TIF

The gut microbiota of wild animals, influenced by various factors including diet, nutrition, gender, and age, plays a critical role in their health and disease status. This study focuses on raccoon dogs (Nyctereutes procyonoides), a commonly found wild animal, and its gut microbiota composition in response to dietary shifts. The study aimed to compare the fecal bacterial communities and diversity of rescued raccoon dogs fed three different diet types (fish and amphibians, mixed protein with maize, and solely maize) using high-throughput sequencing. Results indicated that the dietary composition significantly influenced the gut microbiota, with notable differences in the abundance of several key phyla and genera. The study identified Firmicutes as the dominant phylum in all diet groups, with notable variations in the relative abundances of Bacteroidota, Proteobacteria, and Verrucomicrobiota. Notably, the group solely fed maize exhibited a significant increase in Proteobacteria, potentially linked to dietary fiber and lignin degradation. The genus-level analysis highlighted significant differences, with Lactobacillus and Bifidobacterium responding to dietary shifts. The genus Akkermansia in Verrucomicrobiota can be identified as a marker for assessing the health of the gut and deserves further investigation. Gender-specific differences in the gut microbiota were observed, highlighting the influence of individual variation. Furthermore, the analysis of bacterial functions suggested a connection between diet and host metabolism, emphasizing the need for further research to understand the complex mechanisms underlying the relationship between dietary composition and gut microbiota in wild animals. These findings provide crucial insights into conservation and rescue efforts for wild animals.</p

IL-4 dysregulates microRNAs involved in inflammation, angiogenesis and apoptosis in epidermal keratinocytes

IL-4 plays an important role in atopic dermatitis (AD) pathogenesis by dysregulating many key factors at the transcriptional level. In this study, using microRNA array technique and IL-4 transgenic mice, we demonstrated that IL-4 dysregulates microRNAs involved in inflammation, angiogenesis, lymphoangiogenesis and apoptosis. Of all the 372 common microRNAs examined, 26 and 1 microRNAs are up- and down-regulated, respectively. MicroRNA-101-5p, -122-5p, -142-3p, -204-5p, -335-3p, -376a-3p, -378a-5p, -639 and -9-5p are among the most significantly up-regulated microRNAs. MicroRNA-147a, the only down-regulated one in our study, attenuates TLR induced-inflammatory response. These dysregulated microRNAs may provide post-transcriptional regulation of key genes in AD

Hollow Rodlike MgF₂ with an Ultralow Refractive Index for the Preparation of Multifunctional Antireflective Coatings

Antireflective coatings with superhydrophobic, self-cleaning, and wide-spectrum high-transmittance properties and good mechanical strength have important practical value. In this research, hollow nanorod-like MgF₂ sols with different void volumes were prepared by a template-free solvothermal method to further obtain hollow nanorod-like MgF₂ crystals with an ultralow refractive index of 1.14. Besides, a MgF₂ coating with an adjustable refractive index of 1.10–1.35 was also prepared by the template-free solvothermal method. Then through the combination of base/acid two-step-catalyzed TEOS and hydroxyl modification on the surface of nanosilica spheres, the SiO₂ coating with good mechanical strength, a flat surface, and a refractive index of 1.30–1.45 was obtained. Double-layer broadband antireflective coatings with an average transmittance of 99.6% at 400–1400 nm were designed using the relevant optical theory. After the coating thickness was optimized by the dip-coating method, the double-layer antireflective coatings, whose parameters were consistent with those designed by the theory, were obtained. The bottom layer was a SiO₂ coating with a refractive index of 1.34 and a thickness of 155 nm, and the top layer was a hollow rodlike MgF₂ coating with a refractive index of 1.10 and a thickness of 165 nm. The average transmittance of the obtained MgF₂–SiO₂ antireflective coatings was 99.1% at 400–1400 nm, which was close to the theoretical value. The hydrophobic angle of the coating surface reached 119° at first, and the angle further reached 152° after conducting surface modification by PFOTES. In addition, because the porosity of the coating surface was only 10.7%, the pencil hardness of the coating surface was 5 H and the critical load Lc was 27.05 N. In summary, the obtained antireflective coatings possessed superhydrophobic, self-cleaning, and wide-spectrum high-transmittance properties and good mechanical strength

Formation of Multicomponent Surface Nanodroplets by Solvent Exchange

Multicomponent surface droplets that consist of more than one compound are of great interest for fundamental studies of microwetting, evaporation, and dissolution behaviors, as well as for practical applications in high-throughput screening, microcompartmentalized chemical reactions, and microanalytics. In this work, we study the formation of multicomponent surface nanodroplets from heterogeneous nucleation and growth induced by the process of solvent exchange. In our experiments, as a solution of two oils in their good solvent was displaced by a poor solvent of the oils in the standard solvent exchange, binary droplets of oils were produced on an immersed substrate. The concentration of one oil was constant in the initial solution, whereas the other oil was increased gradually. We characterized the ratio of the two oils inside individual binary droplets by an infrared microspectrometer. Our results show that the ratio of two oils within binary nanodroplets could be varied from 0 to 100% by tuning the composition of the initial solution. However, the ratio of the two oils in the droplets did not simply correspond to that in the solution. Rather, we were able to correlate the ratio of the oils in the droplet to the oversaturation level of each oil based on the ternary phase diagram. We further demonstrate that the principle of the oversaturation level also governs the components in ternary nanodroplets formed by solvent exchange. The quantitative understanding in this work is valuable for the formation of multicomponent surface nanodroplets, which may be applied in nanoextraction, microcompartmentalized reactions, and surface functionalization

Additional file 1: Table S1. of Multipotent luminal mammary cancer stem cells model tumor heterogeneity

Oligonucleotides. Table S2. Confusion matrix for the validation dataset. Table S3. Cell line properties. Table S4. 37 Geneset. (PDF 102 kb

Additional file 4: Figure S3. of Multipotent luminal mammary cancer stem cells model tumor heterogeneity

Py230 cells are estrogen responsive. A. Py230 cells cultured on glass coverslips for 48 hours in F12K/DMEM media with 5 % fetal calf serum. B. Py230 cells cultured on glass coverslips for 48 hours in F12K/DMEM media without phenol red with 5 % charcoal-treated serum. C, D. 104 × Py230 cells injected into intact or OVX female mice were monitored over an eight-week period. Tumor volume and final tumor burden data indicated that the majority of Py230 tumors were sensitive to the presence of endogenous estrogen. Data are means ± SEM from 16 tumors in four mice per group. E,F. Tumors from the same dataset that have undergone epigenetic and/or genetic changes resulting in increased growth (encircled in D) have been removed. Data are means ± SEM from 15 tumors in intact mice and 13 tumors in OVX mice. Data are representative of two similar experiments. (PDF 504 kb

Additional file 2: Figure S1. of Multipotent luminal mammary cancer stem cells model tumor heterogeneity

Clonal MaCSC differentiation. A. Clonal single cells from FVB/N cell line Py9813 are K8K14-positive. Left panel stained with secondary antibody alone. B. Early colonies contain only double-positive cells. Scale bar 20 μm. C. In two-dimensional cultures, K8 + K14+ Py9813 cells can self-renew or differentiate to form single-positive K8 or K14-positive cells. Scale bar 20 μm. D. Py9813 cells form hollow mammospheres that show the presence of double-positive cells and K8 single-positive luminal cells. Single-positive myoepithelial cells are less commonly observed. Scale bar 20 μm. E. The percentage of double-positive, single-positive and double-negative cells in immunofluorescence-stained two-dimensional cultures. Analysis of variance showed the groups were highly significantly different, P <0.0001 and by Holm-Sidak’s multiple comparisons test, there were significantly fewer K8+ and K14+ cells compared with K8 + K14+ cells, P <0.0001, and significantly fewer K14+ cells than K8+ cells, P <0.01 in both cell lines. No double-negative cells were observed in cultures of either cell type. Data are means ± SEM. F. Cultures of control or 300 nM retinoic acid-treated Py9813 cells stained with oil red O. Scale bar 20 μm. (PDF 900 kb

Additional file 5: Figure S4. of Multipotent luminal mammary cancer stem cells model tumor heterogeneity

Validation of expression array data by GSEA and RT-PCR. A. Gene set for mammary luminal progenitor cells is significantly enriched (P <0.001) in luminal versus claudin-low tumors. B. Gene set for mammary stem cells is not enriched in luminal versus claudin-low tumors. C. Genes downregulated in mammary stem cells are significantly enriched (P <0.01) in the Py230 luminal cell line compared with the Py16-1 claudin-low cell line. D. Confirmation of lower expression of several mammary luminal progenitor-associated genes in Py16-1, a claudin-low cell line compared with the parental Py230 cell line. E. Confirmation of higher expression of several mammary stem cell-associated genes in Py16-1, a claudin-low cell line compared with the parental Py230 cell line. F. Higher expression of genes downregulated in mammary stem cells in luminal Py230 cell line compared with claudin-low Py16-1. For each gene comparison *P <0.05, ***P <0.005, ****P <0.001 with Sidak’s correction for multiple comparisons, following two-way analysis of variance. Data are means ± SEM for three independent cultures per group. G. 35 independent tumors were examined for expression of each of the indicated markers. The results were compared with expression by normal prelactating mammary gland (PLMG), which is the normal control for highly proliferative mammary tumors and spontaneous PyVmT tumors (n = 4 per group). (PDF 386 kb