Myofibroblastic cancer-associated fibroblast subtype heterogeneity in pancreatic cancer.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) has a fibrotic stroma that has both tumor-promoting and tumor-restraining properties. Different types of cancer-associated fibroblasts (CAFs) have been described. Here, we investigated whether CAFs within the same subtype exhibit heterogeneous functions. METHODS: We evaluated the gene and protein expression differences in two myofibroblastic CAF (myCAF) lines using single-cell and bulk RNA-sequencing. We utilized proliferation and migration assays to determine the effect of different CAF lines on a tumor cell line. RESULTS: We found that myCAF lines express an activated stroma subtype gene signature, which is associated with a shorter survival in patients. Although both myCAF lines expressed α-smooth muscle actin (α-SMA), platelet-derived growth factor-α (PDGFR-α), fibroblast-activated protein (FAP), and vimentin, we observed heterogeneity between the two lines. Similarly, despite being consistent with myCAF gene expression overall, heterogeneity within specific genes was observed. We found that these differences extended to the functional level where the two myCAF lines had different effects on the same tumor cell line. The myCAF216 line, which had slightly increased inflammatory CAF-like gene expression and higher protein expression of α-SMA, PDGFR-α, and FAP was found to restrain migration of tumor cells. CONCLUSIONS: We found that two myCAF lines with globally similar expression characteristics had different effects on the same tumor cell line, one promoting and the other restraining migration. Our study highlights that there may be unappreciated heterogeneity within CAF subtypes. Further investigation and attention to specific genes or proteins that may drive this heterogeneity will be important.

Antioxidant Effects and Probiotic Properties of Latilactobacillus sakei MS103 Isolated from Sweet Pickled Garlic.

Fermented vegetable-based foods, renowned for their unique flavors and human health benefits, contain probiotic organisms with reported in vitro antioxidative effects. This study investigates the probiotic properties of Latilactobacillus sakei MS103 (L. sakei MS103) and its antioxidant activities using an in vitro oxidative stress model based on the hydrogen peroxide (H2O2)-induced oxidative damage of RAW 264.7 cells. L. sakei MS103 exhibited tolerance to extreme conditions (bile salts, low pH, lysozyme, H2O2), antibiotic sensitivity, and auto-aggregation ability. Moreover, L. sakei MS103 co-aggregated with pathogenic Porphyromonas gingivalis cells, inhibited P. gingivalis-induced biofilm formation, and exhibited robust hydrophobic and electrostatic properties that enabled it to strongly bind to gingival epithelial cells and HT-29 cells for enhanced antioxidant effects. Additionally, L. sakei MS103 exhibited other antioxidant properties, including ion-chelating capability and the ability to effectively scavenge superoxide anion free radicals, hydroxyl, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid, and 2,2-diphenyl-1-picrylhydrazyl. Furthermore, the addition of live or heat-killed L. sakei MS103 cells to H2O2-exposed RAW 264.7 cells alleviated oxidative stress, as reflected by reduced malondialdehyde levels, increased glutathione levels, and the up-regulated expression of four antioxidant-related genes (gshR2, gshR4, Gpx, and npx). These findings highlight L. sakei MS103 as a potential probiotic capable of inhibiting activities of P. gingivalis pathogenic bacteria and mitigating oxidative stress.

Leuconostoc mesenteroides LVBH107 Antibacterial Activity against Porphyromonas gingivalis and Anti-Inflammatory Activity against P. gingivalis Lipopolysaccharide-Stimulated RAW 264.7 Cells.

Probiotics, active microorganisms benefiting human health, currently serve as nutritional supplements and clinical treatments. Periodontitis, a chronic infectious oral disease caused by Porphyromonas gingivalis (P. gingivalis), activates the host immune response to release numerous proinflammatory cytokines. Here, we aimed to clarify Leuconostoc mesenterica (L. mesenteroides) LVBH107 probiotic effects based on the inhibition of P. gingivalis activities while also evaluating the effectiveness of an in vitro P. gingivalis lipopolysaccharide-stimulated RAW 264.7 cell-based inflammation mode. L. mesenteroides LVBH107 survived at acid, bile salts, lysozyme, and hydrogen peroxide conditions, auto-aggregated and co-aggregated with P. gingivalis, exhibited strong hydrophobicity and electrostatic action, and strongly adhered to gingival epithelial and HT-29 cells (thus exhibiting oral tissue adherence and colonization abilities). Moreover, L. mesenteroides LVBH107 exhibited sensitivity to antibiotics erythromycin, doxycycline, minocycline, ampicillin, and others (thus indicating it lacked antibiotic resistance plasmids), effectively inhibited P. gingivalis biofilm formation and inflammation (in vitro inflammation model), reduced the secretion of pro-inflammatory cytokines (TNF-α, IL-6 and IL-1ß) and inflammatory mediators (NO and PGE2), and decreased the expression levels of inflammation related genes. Thus, L. mesenterica LVBH107 holds promise as a probiotic that can inhibit P. gingivalis biofilm formation and exert anti-inflammatory activity to maintain oral health.

Antibacterial and anti-biofilm activities of probiotic Lactobacillus curvatus BSF206 and Pediococcus pentosaceus AC1-2 against Streptococcus mutans.

Streptococcus mutans (S. mutans) is the most important oral pathogenic bacterial cause of dental caries. Here we investigated the abilities of probiotic lactobacillus strains of Lactobacillus curvatus (L. curvatus) BSF206 and Pediococcus pentosaceus (P. pentosaceus) AC1-2 to control S. mutans. Both probiotic strains are acid and bile salt tolerant and are resistant to hydrogen peroxide and lysozyme to promote their survival within the oral environment. In addition, both strains are highly hydrophobic and are also capable of engaging in electrostatic interactions. These properties enhance abilities of both strains to adhere to gingival epithelial cells and HT-29 for improved colonization of oral tissues, while also enabling these probiotics auto-aggregate and to form aggregates with S. mutans that both may prevent S. mutans from colonizing oral tissues and facilitate the clearance of the cariogenic bacteria from the mouth during swallowing of food and saliva. Furthermore, results presented herein revealed that L. curvatus BSF206 and P. pentosaceus AC1-2 effectively inhibited S. mutans activities (biofilm formation, secretion of extracellular matrix components, synthesis of water-insoluble glucans) and led to downregulation of expression of key S. mutans genes involved in biofilm production (gtfA, gtfB, ftf, brpA). Taken together, these results indicate that L. curvatus BSF206 and P. pentosaceus AC1-2 can inhibit S. mutans biofilm formation as a new strategy for preventing dental caries.

Novel insight from the first lung transplant of a COVID-19 patient.

BACKGROUND: To reveal detailed histopathological changes, virus distributions, immunologic properties and multi-omic features caused by SARS-CoV-2 in the explanted lungs from the world's first successful lung transplantation of a COVID-19 patient. MATERIALS AND METHODS: A total of 36 samples were collected from the lungs. Histopathological features and virus distribution were observed by optical microscope and transmission electron microscope (TEM). Immune cells were detected by flow cytometry and immunohistochemistry. Transcriptome and proteome approaches were used to investigate main biological processes involved in COVID-19-associated pulmonary fibrosis. RESULTS: The histopathological changes of the lung tissues were characterized by extensive pulmonary interstitial fibrosis and haemorrhage. Viral particles were observed in the cytoplasm of macrophages. CD3+ CD4- T cells, neutrophils, NK cells, γ/δ T cells and monocytes, but not B cells, were abundant in the lungs. Higher levels of proinflammatory cytokines iNOS, IL-1ß and IL-6 were in the area of mild fibrosis. Multi-omics analyses revealed a total of 126 out of 20,356 significant different transcription and 114 out of 8,493 protein expression in lung samples with mild and severe fibrosis, most of which were related to fibrosis and inflammation. CONCLUSIONS: Our results provide novel insight that the significant neutrophil/ CD3+ CD4- T cell/ macrophage activation leads to cytokine storm and severe fibrosis in the lungs of COVID-19 patient and may contribute to a better understanding of COVID-19 pathogenesis.

Hydrogen Sulfide Attenuates Hydrogen Peroxide-Induced Injury in Human Lung Epithelial A549 Cells.

Lung tissues are frequently exposed to a hyperoxia environment, which leads to oxidative stress injuries. Hydrogen sulfide (H2S) is widely implicated in physiological and pathological processes and its antioxidant effect has attracted much attention. Therefore, in this study, we used hydrogen peroxide (H2O2) as an oxidative damage model to investigate the protective mechanism of H2S in lung injury. Cell death induced by H2O2 treatment could be significantly attenuated by the pre-treatment of H2S, resulting in a decrease in the Bax/Bcl-2 ratio and the inhibition of caspase-3 activity in human lung epithelial cell line A549 cells. Additionally, the results showed that H2S decreased reactive oxygen species (ROS), as well as neutralized the damaging effects of H2O2 in mitochondria energy-producing and cell metabolism. Pre-treatment of H2S also decreased H2O2-induced suppression of endogenous H2S production enzymes, cystathionine-beta-synthase (CBS), cystathionine-gamma-lyase (CSE), and 3-mercapto-pyruvate sulfurtransferase (MPST). Furthermore, the administration of H2S attenuated [Ca2+] overload and endoplasmic reticulum (ER) stress through the mitogen-activated protein kinase (MAPK) signaling pathway. Therefore, H2S might be a potential therapeutic agent for reducing ROS and ER stress-associated apoptosis against H2O2-induced lung injury.

Prebiotic Potential of Xylooligosaccharides Derived from Corn Cobs and Their In Vitro Antioxidant Activity When Combined with Lactobacillus.

In the present work, the in vitro prebiotic activity of xylooligosaccharides (XOS) derived from corn cobs combined with Lactobacillus plantarum, a probiotic microorganism, was determined. These probiotics exhibited different growth characteristics depending on strain specificity. L. plantarum S2 cells were denser and their growth rates were higher when cultured on XOS. Acetate was found to be the major short-chain fatty acid produced as the end-product of fermentation, and its amount varied from 1.50 to 1.78 mg/ml. The antimicrobial activity of XOS combined with L. plantarum S2 was determined against gastrointestinal pathogens. The results showed that XOS proved to be an effective substrate, enhancing antimicrobial activity for L. plantarum S2. In vivo evaluation of the influence of XOS and L. plantarum S2, used both alone and together, on the intestinal microbiota in a mouse model showed that XOS combined with L. plantarum S2 could increase the viable lactobacilli and bifidobacteria in mice feces and decrease the viable Enterococcus, Enterobacter, and Clostridia spp. Furthermore, in the in vitro antioxidant assay, XOS combined with L. plantarum S2 possessed significant 2,2-diphenyl-1- picrylhydrazyl, 2,2'-azino-bis, and superoxide anion radical-scavenging activities, and the combinations showed better antioxidant activity than either XOS or L. plantarum S2 alone.

2-Amino-6-(naphthalen-1-yl)-4-phenyl-pyridine-3-carbonitrile.

In the title compound, C(22)H(15)N(3), the naphthyl ring system makes dihedral angles of 67.40 (2) and 59.80 (3)° with the pyridyl and phenyl rings, respectively. In the crystal, the mol-ecules are connected via inter-molecular N-H⋯N hydrogen bonds, forming a three-dimensional network.

5-(4-Methyl-piperazin-1-yl)-2-nitro-aniline.

In the title compound, C(11)H(16)N(4)O(2), the dihedral angle between the benzene ring and the plane of the four carbon atoms in the piperazine ring is 12.17 (3)°; the latter ring adopts a chair conformation. An intramolecular N-H⋯O hydrogen bond generates an S(6) ring. In the crystal, the molecules are linked by N-H⋯N hydrogen bonds, forming chains.

Poly[di-µ(2)-chlorido-µ(2)-(1,4-dioxane-κO:O')-cadmium(II)].

In the title complex, [CdCl(2)(C(4)H(8)O(2))](n), two different Cd(II) ions are present, one in a general position and one with site symmetry 2. The Cd(II) ions are coordinated by two O atoms from two 1,4-dioxane ligands and four chloride anions in a slightly distorted octa-hedral geometry and is connected to neighboring Cd(II) ions by two bridging chloride anions, generating infinite linear chains along the a axis. These chains are further inter-connected by bridging 1,4-dioxane ligands, affording a three-dimensional network.

2-Amino-4-(2-fluoro-phen-yl)-5,6-dihydro-benzo[h]quinoline-3-carbonitrile.

In the title compound, C(20)H(14)FN(3), the F atom of the fluoro-substituted benzene ring in the 4-position of the 5,6-dihydro-benzo[h]quinoline system is disordered over two positions (0.80 and 0.20 occupancy). The dihedral angle between the pyridine and fluorobenzene rings is 73.2 (2) Å. The crystal structure is established by inter-molecular N-H⋯N hydrogen bonds, forming a three-dimensional network.

Specific renal uptake of randomly 50% N-acetylated low molecular weight chitosan.

In our previous studies, randomly 50% N-acetylated low molecular weight chitosan (LMWC) has been confirmed as a potential carrier for the site-specific delivery of prednisolone to kidney, suggesting specific uptake of LMWC in kidney. Interestingly, aminoglycoside, a well-known ligand of megalin receptor, shares a similar glucosamine unit level with LMWC. Based on these, we proposed that the specific renal uptake of LMWC might also be mediated by the megalin receptor. To test this hypothesis, we performed the present study to further investigate the renal uptake process of LMWC and its possible mechanism as well. First, LMWC was found by fluorescent microscopy to selectively accumulate in the kidneys, especially in the renal proximal tubules after iv injection in mice. Then, our research also revealed that LMWC was internalized into renal tubular cells (RTCs) through endocytic pathway, with a concentration-dependent and saturable pattern. The uptake of LMWC could be competitively inhibited in the presence of gentamycin, a kind of aminoglycosides. In addition, cytotoxicity assay showed that there were no obvious effects of LMWC on the viability of L929 and RTCs lines. Finally, megalin-shedding mouse models were established and the distribution of LMWC in tissues of normal and megalin-shedding mice was evaluated. Consistent with gentamycin inhibition assay, in vivo results also suggested the role of megalin in the uptake of LMWC in kidney. In conclusion, LMWC could be specifically taken up by RTCs, where the megalin receptor would likely mediate its binding and uptake.