Endometrial Epithelial ARID1A Is Required for Uterine Immune Homeostasis during Early Pregnancy

A growing body of work suggests epigenetic dysregulation contributes to endometriosis pathophysiology and female infertility. The chromatin remodeling complex subunit AT-rich interaction domain 1A (ARID1A) must be properly expressed to maintain normal uterine function. Endometrial epithelial ARID1A is indispensable for pregnancy establishment in mice through regulation of endometrial gland function; however, ARID1A expression is decreased in infertile women with endometriosis. We hypothesized that ARID1A performs critical operations in the endometrial epithelium necessary for fertility besides maintaining gland function. To identify alterations in uterine gene expression resulting from loss of epithelial ARID1A, we performed RNA-sequencing analysis on pre-implantation uteri from Ltf^iCre/+Arid1a^f/f and control mice. Differential expression analysis identified 4181 differentially expressed genes enriched for immune-related ingenuity canonical pathways including agranulocyte adhesion and diapedesis and natural killer cell signaling. RT-qPCR confirmed an increase in pro-inflammatory cytokine and macrophage-related gene expression but a decrease in natural killer cell signaling. Immunostaining confirmed a uterus-specific increase in macrophage infiltration. Flow cytometry delineated an increase in inflammatory macrophages and a decrease in uterine dendritic cells in Ltf^iCre/+Arid1a^f/f uteri. These findings demonstrate a role for endometrial epithelial ARID1A in suppressing inflammation and maintaining uterine immune homeostasis, which are required for successful pregnancy and gynecological health.     

Generalized Approach towards Secretion-Based Protein Production via Neutralization of Secretion-Preventing Cationic Substrate Residues

Many heterologous proteins can be secreted by bacterial ATP-binding cassette (ABC) transporters, provided that they are fused with the C-terminal signal sequence, but some proteins are not secretable even though they carry the right signal sequence. The invention of a method to secrete these non-secretable proteins would be valuable both for understanding the secretory physiology of ABC transporters and for industrial applications. Herein, we postulate that cationic “supercharged” regions within the target substrate protein block the secretion by ABC transporters. We also suggest that the secretion of such substrate proteins can be rescued by neutralizing those cationic supercharged regions via structure-preserving point mutageneses. Surface-protruding, non-structural cationic amino acids within the cationic supercharged regions were replaced by anionic or neutral hydrophilic amino acids, reducing the cationic charge density. The examples of rescued secretions we provide include the spike protein of SARS-CoV-2, glutathione-S-transferase, streptavidin, lipase, tyrosinase, cutinase, growth factors, etc. In summary, our study provides a method to predict the secretability and a tool to rescue the secretion by correcting the secretion-blocking regions, making a significant step in understanding the physiological properties of ABC transporter-dependent protein secretion and laying the foundation for the development of a secretion-based protein-producing platform.     

Intratracheal Transplantation of Mesenchymal Stem Cells Attenuates Hyperoxia-Induced Microbial Dysbiosis in the Lungs,Brain, and Gut in Newborn Rats

We attempted to determine whether intratracheal (IT) transplantation of mesenchymal stem cells (MSCs) could simultaneously attenuate hyperoxia-induced lung injuries and microbial dysbiosis of the lungs, brain, and gut in newborn rats. Newborn rats were exposed to hyperoxia (90% oxygen) for 14 days. Human umbilical cord blood-derived MSCs (5 × 10⁵) were transplanted via the IT route on postnatal day (P) five. At P14, the lungs were harvested for histological, biochemical, and microbiome analyses. Bacterial 16S ribosomal RNA genes from the lungs, brain, and large intestine were amplified, pyrosequenced, and analyzed. IT transplantation of MSCs simultaneously attenuated hyperoxia-induced lung inflammation and the ensuing injuries, as well as the dysbiosis of the lungs, brain, and gut. In correlation analyses, lung interleukin-6 (IL-6) levels were significantly positively correlated with the abundance of Proteobacteria in the lungs, brain, and gut, and it was significantly inversely correlated with the abundance of Firmicutes in the gut and lungs and that of Bacteroidetes in the lungs. In conclusion, microbial dysbiosis in the lungs, brain, and gut does not cause but is caused by hyperoxic lung inflammation and ensuing injuries, and IT transplantation of MSCs attenuates dysbiosis in the lungs, brain, and gut, primarily by their anti-oxidative and anti-inflammatory effects.     

Prognosis of breast cancer is associated with one-carbon metabolism related nutrients among Korean women

ABSTRACT: BACKGROUND: The 5-year survival rate for breast cancer among Korean women has increased steadily; however, breast cancer remains the leading cause of cancer mortality among women. One-carbon metabolism, which requires an adequate supply of methyl group donors and B vitamins, may affect the prognosis of breast cancer. This aim of this study was to investigate the associations of dietary intake of vitamin B2, vitamin B6 and folate before diagnosis on the prognosis of breast cancer. METHODS: We assessed the dietary intake using a food frequency questionnaire with 980 women who were newly diagnosed and histopathologically confirmed to have primary breast cancer from hospitals in Korea, and 141 disease progression events occurred. Cox's proportional hazard regression models were used to estimate the hazard ratio (HR) and 95% confidence interval (95% CI) adjusting for age, education, recruitment sites, TNM stage, hormone status, nuclear grade and total calorie. RESULTS: There was no significant association between any one-carbon metabolism related nutrients (vitamin B2, B6 and folate) and the progression of breast cancer overall. However, one-carbon metabolism related nutrients were associated with disease progression in breast cancer patients stratified by subtypes. In ER + and/or PR + breast cancers, no association was observed; however, in ER--/PR-- breast cancers, a high intake of vitamin B2 and folate statistically elevated the HR of breast cancer progression (HR = 2.28; 95% CI, 1.20-4.35, HR = 1.84; 95% CI, 1.02-3.32, respectively) compared to a low intake. This positive association between the ER/PR status and progression of the disease was profound when the nutrient intakes were categorized in a combined score (Pinteraction = 0.018). In ER--/PR-- breast cancers, high combined scores were associated with a significantly poor DFS compared to those belonging to the low score group (HR = 3.84; 95% CI, 1.70-8.71). CONCLUSIONS: In conclusion, our results suggest that one-carbon related nutrients have a role in the prognosis of breast cancer depending on the ER/PR status.     

An efficient microwave-assisted Suzuki reaction using Pd/MCM-41 and Pd/SBA-15 as catalysts in solvent-free condition

Palladium catalysts, Pd/MCM-41 and Pd/SBA-15 were prepared by impregnation of an aqueous solution of [Pd(NH₃)₄]Cl₂ on MCM-41 and SBA-15. Palladium contents of Pd/MCM-41 and Pd/SBA-15 are 8.4% and 8.7%, respectively. It has been shown that these catalysts are very suitable to microwave-assisted Suzuki reactions under solvent-free condition. It is also found that the base additives for this reaction are K₂CO₃, Cs₂CO₃ or CsF. Thus, phenylboronic acid and phenyl iodide with Pd/MCM-41 produce biphenyl by microwave irradiation for 10 min in 97.4% yield. Phenyl bromide, instead of phenyl iodide, also proceeds the reaction with phenylboronic acid using Pd/MCM-41 or Pd/SBA-15 yielding biphenyl by microwave irradiation for 10 min in excellent yield. Whereas the reaction of phenyl chloride with phenylboronic acid gives poor yield in same condition. Various aryl iodides and aryl bromides are tested. In this paper our recent results of microwave-assisted Suzuki reaction using Pd/MCM-41 and Pd/SBA-15 under solvent-free condition are described.     

Potential Chemical Markers for the Identification of Irradiated Sausages

Abstract: Hydrocarbons, gas compounds, and off‐odor volatiles were determined for irradiated (0 or 5 kGy) commercial sausages with different fat contents (16% and 29%) during a 60‐d storage period at 4 °C. Total of 4 hydrocarbons (C14:1, C15:0, C16:2, and C17:1) were detected only in irradiated sausages: the amount of C16:2 was the highest, followed by C17:1, C14:1, and C15:0. The concentrations of hydrocarbons decreased significantly (P < 0.05) with storage, but were still detectable at the end of 60‐d storage. Irradiated sausages produced significantly higher amounts of CO than the nonirradiated ones. CH₄ was detected only in irradiated sausages. Dimethyl disulfide was detected only in irradiated sausages and its concentration decreased significantly (P < 0.05) with storage. Fat content of sausages showed a significant effect on the production and retention of hydrocarbons, gas compounds, and sulfur volatiles in irradiated sausages during storage. Some hydrocarbons (C16:2, C17:1, C14:1, and C15:0), CH₄, and dimethyl disulfide were only found in irradiated sausages indicating that these compounds can be used as potential markers for irradiated sausages.     

Molecular brushes with extreme grafted side chain densities

A series of densely grafted poly(n-butyl acrylate) (PBA) molecular brushes with four different grafting densities were synthesized by the “grafting-from” approach using atom transfer radical polymerization (ATRP). A novel monomer, isopropylidene-2,2-Bis(methoxy)propionic hydroxyethylmethacrylate (IMPHMA), was synthesized and copolymerized with methyl methacrylate (MMA) under different monomer feed ratios to yield a series of linear poly(methyl methacrylate-stat-IMAPA), [PMMA-s-(PIMPHMA)]. The resulting copolymers were deprotected and transformed to macroinitiators, [PMMA-s-(PHEMA-IMPHMA-Br)]. n-Butyl acrylate (BA) was grafted from these macroinitiators to yield a series of molecular brushes, [PMMA-s-{(PIMPHMA)-g-PBA}], with various side chain lengths. Molecular brushes were characterized by gel permeation chromatography (GPC) and ¹H NMR. PBA side chains were cleaved by acid hydrolysis, and the resulting linear PBA polymers were characterized by GPC to study initiation efficiency during the synthesis of molecular brushes. The initiation efficiency increased with polymerization time and decreased with macroinitiators that had more initiation sites. Atomic force microscopy (AFM) measurements demonstrated the characteristic molecular structure by resolving individual brush molecules.