Journal: bioRxiv

Article Title: Antimicrobial peptides modulate lung injury by altering the intestinal microbiota

doi: 10.1101/2023.03.14.529700

Figure Lengend Snippet: A) Neonatal C57BL/6J mice were exposed to normoxia or hyperoxia from the 3 rd -14 th day of life (n = 4 litters with 5-7 neonatal mice/litter per exposure group). FiO 2 , fraction of inspired oxygen. SPF, specific-pathogen-free. B) Representative photomicrographs of the distal lung sections of 14-day-old mice. C) Hyperoxia exposure is associated with alterations in lung morphology and function. Data are shown as mean ± SEM, with significance testing by two-tailed t -test. D) Volcano plot of ileal gene expression array showing gene expression altered by hyperoxia exposure. E) Heatmap showing genes regulated by hyperoxia exposure. F) Principal components analysis showing differential clustering of normoxia and hyperoxia exposed ileal genes. PC, principal component. G) Ileal antimicrobial peptide expression is decreased in hyperoxia-exposure mice. H) Community diversity of the adherent and luminal ileal bacterial microbiome is not significantly altered by hyperoxia exposure. I) The relative abundance of an operational taxonomic units (OTU 002) that aligns to the genus Staphylococcus increases after hyperoxia exposure, as do OTUs aligning to Corynebacterium (OTU 124) and Romboutsia (OTU 013). Data are shown as mean ± SEM, with significance testing by two-tailed t -test. J) Principal coordinates analysis of Bray-Curtis dissimilarity show global alterations in community composition in hyperoxia-exposed mice. Significance testing by permutational ANOVA (PERMANOVA), with equivocal dispersion confirmed by permutational multivariate analysis of dispersion (PERMDISP). PC, principal component. K) Loading plot of principal components analysis of Hellinger transformed Euclidian distances showing the contribution of specific genera to the global community composition. Schematic in (A) was generated using BioRender. See also and .

Article Snippet: In the first experiment, we used 8-week-old female C57Bl/6J mice (Stock Number 000664) purchased from the Jackson Laboratory (hence, JAX) and allowed to acclimate in specific-pathogen free (SPF) housing with 4 animals per cage, on a 12-hour light/dark cycle, at 20-24C, with ad libitum access to irradiated rodent chow (Teklad 7904, Envigo) in SPF Vivarium 1 (Pittman Biomedical Research Building II) for two weeks.

Techniques: Two Tailed Test, Expressing, Transformation Assay, Generated

Journal: bioRxiv

Article Title: Antimicrobial peptides modulate lung injury by altering the intestinal microbiota

doi: 10.1101/2023.03.14.529700

Figure Lengend Snippet: A) Small intestinal spheroid organoids derived from neonatal C57BL/6J mice were exposed to either hyperoxia or normoxia for 24 hours. FiO 2 , fraction of inspired oxygen. B) Representative images of organoids before and after exposure, with insets at 40x magnification. The percentage of organoids with healthy appearing epithelium declined in hyperoxia-exposed organoids. Data are shown as mean ± SEM, with significance testing by a two-tailed t -test. Scale bar represents 1000 μm. C) Representative immunohistochemistry after exposure to normoxia or hyperoxia. Nuclei in blue, actively proliferating cells in green, and lysozyme-positive cells in red. Arrows identify lysozyme-positive Paneth cells. Scale bar represents 25 μm. D) Principal components analysis showing differential clustering of normoxia and hyperoxia exposed ileal genes. PC, principal component. E) Heatmap showing genes regulated by hyperoxia exposure. F) Heatmap of antimicrobial peptide expression is decreased in hyperoxia-exposure organoids. G) Ingenuity pathway analysis showing regulated pathways in hyperoxia or normoxia. H) Bubble plot showing up and down-regulated pathways from hyperoxia exposure. The schematic in (A) was generated using BioRender.

Article Snippet: In the first experiment, we used 8-week-old female C57Bl/6J mice (Stock Number 000664) purchased from the Jackson Laboratory (hence, JAX) and allowed to acclimate in specific-pathogen free (SPF) housing with 4 animals per cage, on a 12-hour light/dark cycle, at 20-24C, with ad libitum access to irradiated rodent chow (Teklad 7904, Envigo) in SPF Vivarium 1 (Pittman Biomedical Research Building II) for two weeks.

Techniques: Derivative Assay, Two Tailed Test, Immunohistochemistry, Expressing, Generated

Journal: bioRxiv

Article Title: Antimicrobial peptides modulate lung injury by altering the intestinal microbiota

doi: 10.1101/2023.03.14.529700

Figure Lengend Snippet: A) Neonatal C57BL/6NCrl mice randomized to either every other day exposure to lysozyme by gastric gavage or their littermate controls were then exposed to normoxia or hyperoxia from the 3 rd -14 th day of life (n = 4 litters with 5-7 neonatal mice/litter per exposure group). FiO 2 , fraction of inspired oxygen. PBS, phosphate-buffered saline (vehicle). SPF, specific-pathogen-free. B) Representative photomicrographs of the distal lung sections of 14-day-old mice. C) Lysozyme exposure ameliorates hyperoxia-induced disruptions in lung morphology and function. Data are shown as mean ±SEM, with significance testing by two-way ANOVA. D) Volcano plot of ileal RNAseq showing gene expression altered by lysozyme exposure. E) Heatmap showing genes regulated by lysozyme exposure. F) Principal components analysis showing differential clustering of ileal genes in lysozyme exposed mice. PC, principal component. G) Ileal antimicrobial peptide expression is altered in lysozyme-exposed mice. H) The community diversity of the adherent and luminal ileal bacterial microbiome is not significantly altered by lysozyme exposure. I) The hyperoxia-induced increase in the relative abundance of operational taxonomic unit 014 ( Staphylococcus ) is ameliorated by lysozyme exposure. Multiple other genera are increased in lysozyme and hyperoxia-exposed mice. Data are shown as mean ± SEM, with significance testing by two-way ANOVA. J) Principal coordinates analysis of Bray-Curtis dissimilarity show global alterations in community composition in lysozyme exposed mice. Significance testing by permutational ANOVA (PERMANOVA), with equivocal dispersion confirmed by permutational multivariate analysis of dispersion (PERMDISP). PC, principal component. K) Loading plot of a principal components analysis of a Hellinger transformed Euclidian distance showing global community composition significantly altered in lysozyme-exposed mice. The schematic in (A) was generated using BioRender. See also .

Article Snippet: In the first experiment, we used 8-week-old female C57Bl/6J mice (Stock Number 000664) purchased from the Jackson Laboratory (hence, JAX) and allowed to acclimate in specific-pathogen free (SPF) housing with 4 animals per cage, on a 12-hour light/dark cycle, at 20-24C, with ad libitum access to irradiated rodent chow (Teklad 7904, Envigo) in SPF Vivarium 1 (Pittman Biomedical Research Building II) for two weeks.

Techniques: Expressing, Transformation Assay, Generated