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1.
Hypertens Res ; 47(6): 1642-1653, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38503939

ABSTRACT

Akkermansia muciniphila (Am) shows a beneficial role as a probiotic in the treatment of metabolic syndrome. However, the mechanism remains to be elucidated. We tested the hypothesis that Am extracellular vesicles (AmEVs) have a protective effect against hypertension. Extracellular vesicles purified from anaerobically cultured Am were characterized by nanoparticle tracking analysis, transmission electron microscopy, and silver stain after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). AmEVs (1.0 × 1010 log particles/L) or vehicles were added into organ baths to induce vasorelaxation. In addition, AmEVs (1.0 × 108 or 1.0 × 109 particles/kg) or vehicles were injected into the tail veins of Wistar-Kyoto rats (WKYs) and spontaneously hypertensive rats (SHRs) weekly for 4 weeks. Peripheral blood mononuclear cells (PBMCs) and splenocytes isolated from both rat strains were analyzed by flow cytometry, RT-qPCR, and western blot. AmEVs affected neither vascular contraction nor endothelial relaxation in thoracic aortas. Moreover, AmEVs protected against the development of hypertension in SHRs without a serious adverse reaction. Additionally, AmEVs increased the population of T regulatory (Treg) cells and tended to reduce proinflammatory cytokines. These results indicate that AmEVs have a protective effect against hypertension without a serious adverse reaction. Therefore, it is foreseen that AmEVs may be utilized as a novel therapeutic for the treatment of hypertension.


Subject(s)
Akkermansia , Extracellular Vesicles , Hypertension , Probiotics , Rats, Inbred SHR , Rats, Inbred WKY , Animals , Male , Rats , Aorta, Thoracic , Leukocytes, Mononuclear , Blood Pressure , Vasodilation , Spleen
2.
Diagnostics (Basel) ; 11(4)2021 Mar 30.
Article in English | MEDLINE | ID: mdl-33808296

ABSTRACT

Cancer screening and diagnosis can be achieved by analyzing specific molecules within serum-derived extracellular vesicles (EVs). This study sought to profile EV-derived proteins to identify potential lung cancer biomarkers. EVs were isolated from 80 serum samples from healthy individuals and cancer patients via polyethylene glycol (PEG)-based precipitation and immunoaffinity separation using antibodies against CD9, CD63, CD81, and EpCAM. Proteomic analysis was performed using 2-D gel electrophoresis and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The expression of proteins that were differentially upregulated in the EVs or tissue of lung cancer samples was validated by Western blotting. The area under the curve (AUC) was calculated to assess the predictability of each differentially expressed protein (DEP) for lung cancer. A total of 55 upregulated protein spots were selected, seven of which (CD5L, CLEC3B, ITIH4, SERFINF1, SAA4, SERFINC1, and C20ORF3) were found to be expressed at high levels in patient-derived EVs by Western blotting. Meanwhile, only the expression of EV CD5L correlated with that in cancer tissues. CD5L also demonstrated the highest AUC value (0.943) and was found to be the core regulator in a pathway related to cell dysfunction. Cumulatively, these results show that EV-derived CD5L may represent a potential biomarker-detected via a liquid biopsy-for the noninvasive diagnosis of lung cancer.

3.
J Hazard Mater ; 329: 280-289, 2017 May 05.
Article in English | MEDLINE | ID: mdl-28183017

ABSTRACT

Arsenic (As) biogeochemistry coupled with iron (Fe) and sulfur (S) was studied using columns packed with As(V)-contaminated sediments under two phases: a reduction phase followed by an oxidation phase. During the reduction phase, four identical columns inoculated with G. sulfurreducens were stimulated with 3mM acetate for 60days. The As(III) in the effluent rapidly increased then gradually decreased. The Fe(II) and sulfate concentration indicated ferrous sulfide precipitation inside the column after day 14 and X-ray absorption near edge structure spectra showed that As(III) was enriched at the column outlet. The genera Desulfosporosinus and Anaeromyxobacter as well as the Geobacter inoculum played a primary role in As reduction. During the oxidation phase, dissolved oxygen was consumed by heterotrophic aerobes belonging to the phylum Cloroflexi in the column with acetate, resulting in more As in the effluent. When only nitrate was injected, sulfur-oxidizing bacteria such as Thiobacillus thioparus instantly oxidized the sulfide formed during the first phase, resulting in less As(V) in the aqueous phase compared to the column with dissolved oxygen alone. This study showed that redox gradients and dynamics linked to Fe and S biogeochemistry have an important role in controlling As mobility in subsurface environments.


Subject(s)
Arsenic/metabolism , Groundwater/microbiology , Iron/metabolism , Microbiota , Sulfur/metabolism , Biotransformation , Firmicutes/metabolism , Geobacter/metabolism , Geologic Sediments/chemistry , Geologic Sediments/microbiology , Groundwater/chemistry , Oxidation-Reduction
4.
J Infect Dis ; 213(1): 131-8, 2016 Jan 01.
Article in English | MEDLINE | ID: mdl-26136468

ABSTRACT

RtxA1 toxin, which results in cytoskeletal rearrangement, contact cytotoxicity, hemolysis, tissue invasion, and lethality in mice, is the most potent cytotoxic virulence factor of Vibrio vulnificus. Bioinformatics analysis of rtxA1 predicted 4 functional domains that presumably performed discrete functions during host cell killing. V. vulnificus RtxA1 has a unique domain designated as RtxA1-D2, corresponding to amino acids 1951-2574, which is absent in Vibrio cholerae multifunctional-autoprocessing repeats-in-toxin, suggesting that this domain confers specific biological functions to V. vulnificus RtxA1. HeLa cells expressing green fluorescent protein-RtxA1-D2 became round and lost their viability. A yeast 2-hybrid system identified prohibitin (PHB) 1 as the host partner of RtxA1-D2. The specific interaction of RtxA1-D2 with PHB1 was confirmed by performing immunoprecipitation. Interestingly, V. vulnificus RtxA1 up-regulated PHB1 expression on the cytoplasmic membrane of host cells. Extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways were confirmed as being important in the up-regulation of PHB1 by using inhibitors. Down-regulation of PHB1 by small interfering RNAs decreased the cytotoxicity of RtxA1-D2 against HeLa cells. The pretreatment of an anti-PHB1 antibody impaired the cytotoxicity of V. vulnificus RtxA1. These results suggest that the involvement PHB1 in the RtxA1 cytotoxicity has significant implications for the pathogenesis of V. vulnificus infections.


Subject(s)
Bacterial Proteins/metabolism , Bacterial Toxins/metabolism , Cell Survival/drug effects , Repressor Proteins/metabolism , Vibrio vulnificus/metabolism , Animals , Antibodies, Bacterial , Bacterial Proteins/chemistry , Bacterial Proteins/pharmacology , Bacterial Toxins/chemistry , Bacterial Toxins/pharmacology , Female , HeLa Cells , Humans , MAP Kinase Signaling System , Mice , Prohibitins , Protein Binding , Repressor Proteins/chemistry , Repressor Proteins/pharmacology , Vibrio vulnificus/chemistry , Vibrio vulnificus/pathogenicity
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