Josh Monts

Neutrophil extracellular traps (NETs) are important components of innate immunity. Neonatal neutrophils (polymorphonuclear leukocytes [PMNs]) fail to form NETs due to circulating NET-inhibitory peptides (NIPs), cleavage fragments of α1-antitrypsin (A1AT). How fetal and neonatal blood NIPs are generated remains unknown, however. The placenta expresses high-temperature requirement serine protease A1 (HTRA1) during fetal development, which can cleave A1AT. We hypothesized that placentally expressed HTRA1 regulates the formation of NIPs and that NET competency changed in PMNs isolated from neonatal HTRA1 knockout mice (HTRA1−/−). We found that umbilical cord blood plasma has elevated HTRA1 levels compared with adult plasma and that recombinant and placenta-eluted HTRA1 cleaves A1AT to generate an A1AT cleavage fragment (A1ATM383S-CF) of molecular weight similar to previously identified NIPs that block NET formation by adult neutrophils. We showed that neonatal mouse pup plasma contains ...

Additional file 2: Figure S2. Related to Fig. 1h. A The p53 CTD (Lys370 – Asp391) in green is superposed on the box3-peptide of PGC1α (Gln203 – Asp224) shown in red. The ERRα LBD is shown in yellow. B The box3-peptide of PGC1α is removed from the complex shown in (A). C Zoomed in the interface between the p53 CTD and the box3-peptide of PGC1α showing three potential non-bonding interactions. D Electrostatic surface representation of ERRα at the binding interface with p53, where the red color indicating negatively charged region, blue color being the positively charged region, and the in-between gray color being the hydrophobic region. Here, the hydrophobic residue Leu383 from p53 is trapped within a hydrophobic pocket formed by several hydrophobic residues of ERRα at the binding interface. E Sequence alignment between the ERα LBD (Met192-Tyr389) and the ERRα LBD (Val225-Tyr414). F Superposition of the ERα LBD (in orange) and the ERRα LBD (in green). G Sequence alignment between the ...

Additional file 4: Figure S4. Related to Fig. 3 A HCT-116p53+/+ cells were treated for 48 h with XCT790 (15 μM) or vehicle (DMSO) and transiently transfected with pCMV flag ERRα or pcDNA3 empty vector (mock). IB analysis was conducted with anti-ERRα, anti-p53, and anti-actin. B-C Enriched KEGG pathways up-regulated and down-regulated obtained by STRING analysis of the membrane/organelle purified proteins fraction comparing (ii) absence of ERRα with (i) presence of ERRα and p53 or comparing (iii) absence of p53 with (i) presence of ERRα and p53. Comparisons between groups were made using multiple t tests with a False Discovery Rate of p ≤ 0.05.

Additional file 1: Figure S1. Related to Fig. 1. A Purified ERRα proteins (100 μL) were loaded onto a Superose 6 size exclusion column. The elution profiles were recorded as absorbance at 280 nm. All the fractions from A7 to B10 were eluted from the column. B All fractions were loaded into an SDS–PAGE and then stained with Coomassie Brilliant Blue R-250 solution. C Western blot analysis was performed using anti-ERRα and anti-p53. D 293T cell line (left) and DLD-1 (right) colon cancer cell line were used to perform endogenous IP followed by IB analysis with anti-ERRα, anti-PGC-1α, and anti-GAPDH.

Helicobacter pylori infection and alcohol intake are independent risk factors in gastric carcinogenesis; however, until now, the combined effect of H. pylori infection and alcohol consumption and the specific mechanism is still problematic. Here, we developed a series of mouse models that progress from chronic gastritis to gastric cancer, induced by infecting H. pylori combined with chronic alcohol consumption and then determining the molecular mechanism of the progression by flow cytometry, western blotting, qPCR, Mito Traker assay in the gastric cancer and T-cell lines. Interleukin-10 (IL-10) knockout mice was used to determine whether IL-10 deficiency directly contributes to H. pylori and alcohol induced gastric tumorigenesis. Alcohol consumption, together with H. pylori infection, causes gastric cancer; IL-10 downregulation and mitochondrial metabolic dysfunction in CD8+ cells are also involved. IL-10 knockout accelerates tumor development in mice with either H. pylori infection...

The COVID‐19 pandemic has brought biosafety to the forefront of many life sciences. The outbreak has compelled research institutions to re‐evaluate biosafety practices and potential at‐risk areas within research laboratories and more specifically within Shared Resource Laboratories (SRLs). In flow cytometry facilities, biological safety assessment encompasses known hazards based on the biological sample and associated risk group, as well as potential or unknown hazards, such as aerosol generation and instrument “failure modes.” Cell sorting procedures undergo clearly defined biological safety assessments and adhere to well‐established biosafety guidelines that help to protect SRL staff and users against aerosol exposure. Conversely, benchtop analyzers are considered low risk due to their low sample pressure and enclosed fluidic systems, although there is little empirical evidence to support this assumption of low risk. To investigate this, we evaluated several regions on analyzers u...

Gastric cancer risk evolves over time due to environmental, dietary, and lifestyle changes, including Helicobacter pylori (H. pylori) infection and consumption of hot peppers (i.e., capsaicin). H. pylori infection promotes gastric mucosal injury in the early phase of capsaicin exposure. This relationship suggests a need to investigate the mechanism of how both H. pylori infection and capsaicin contribute to gastric inflammation and lead to gastric cancer. C57-Balb/c mice were infected with the H. pylori (SS1) strain and then fed capsaicin (0.05% or 0.2 g/kg/day) or not. Consequently, tumor size and phenotype were analyzed to determine the molecular mechanism driving the shift from gastritis to stomach cancer. Moreover, we used 2-difluoromethylornithine (DFMO) in mice to prevent gastric tumorigenesis by reducing inflammation and promoting recovery of disease-free stasis. This study provides evidence showing that a combination of H. pylori infection and capsaicin consumption leads to ...