PMID: 

Ann Palliat Med. 2020 May 7. Epub 2020 May 7. PMID: 32389004

Abstract Title: 

Blueberry polyphenols play a preventive effect on alcoholic fatty liver disease C57BL/6 J mice by promoting autophagy to accelerate lipolysis to eliminate excessive TG accumulation in hepatocytes.

Abstract: 

BACKGROUND: This study aims to explore blueberry polyphenols and its roles in nonalcoholic fatty liver disease (NAFLD) by relieving hepatic steatosis, and to understand alcoholic fatty liver disease (AFLD). Cell autophagy has been proved to promote lipid metabolism and is involved in the pathogenesis of AFLD; however, whether blueberry polyphenol affects autophagy is unknown. Therefore, our study analyzes the functions of blueberry polyphenol on AFLD and if its mechanisms are engaged with hepatocytes autophagy.METHODS: We built the AFLD mice model via alcohol abduction, and the TG lipid droplets content detected the hepatic steatosis through ORO and HE stains. For blood lipid levels measurements, serum CHOL and TG concentrations were tests. For mechanism analysis, the lipogenic genes of SREBP1, FAS, and ACCα, and the lipodieretic genes of ATGL and Sirt1 were evaluated by qRT-PCR, as well as the autophagy proteins of p62; WB measured LC3-I and LC3-II.RESULTS: We found that chronic alcohol intake successfully induced AFLD occurrence with increased TG lipid droplets content in liver and serum CHOL and TG levels that accompanied by increased lipogenic and reduced lipodieretic mRNA levels, as well as enhancive p62 protein and decreased LC3-II/LC3-I proportion. However, after blueberry polyphenol intake, there were opposite outcomes happened. Moreover, blueberry polyphenol alone did not affect the lipid metabolism but promoted the hepatocytes autophagy at 200 mg/kg concentration.CONCLUSIONS: In summary, we are unparalleled that illustrated blueberry polyphenols can prevent AFLD development by promoting autophagy to accelerate lipid metabolism than to lighten hepatic steatosis.

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PMID: 

Biomolecules. 2020 May 10 ;10(5). Epub 2020 May 10. PMID: 32397678

Abstract Title: 

Blueberry-Derived Exosome-Like Nanoparticles Counter the Response to TNF-α-Induced Change on Gene Expression in EA.hy926 Cells.

Abstract: 

Exosome-like nanoparticles (ELNs) are attracting interest as important vehicles of intercellular communication, both in prokaryotes and eukaryotes. Recently, dietary nanoparticles similar to mammalian exosomes have attracted attention for these features. In particular they appear to be relevant in the modulation of several cellular processes as well as candidate carriers of bioactive molecules (proteins, lipids, and nucleic acids, including miRNAs) with therapeutic value. Herein, we investigated the cellular uptake of blueberry-derived ELNs (B-ELNs) by a human stabilized endothelial cell line (EA.hy926) and the ability of B-ELNs to modulate the expression of inflammatory genes as the response of tumor necrosis factor-α (TNF-α). Our results indicate that 1) EA.hy926 cells internalize B-ELNs in a dose-dependent manner; 2) pretreatment with B-ELNs counters TNF-α-induced reactive oxygen species (ROS) generation and loss of cell viability and modulates the differential expression of 29 genes (fold change>1.5) induced by TNF-α compared to control; 3) pathway analysis reveals their involvement in a total of 340 canonical pathways, 121 KEGG pathways, and 121 GO Biological processes; and 4) the intersection between differentially expressed (DE) genes and miRNAs contained in B-ELNs unveils a set of candidate target genes,such as prostaglandin I2 synthase (PTGIS), mitogen-activated protein kinase 14 (MAPK14), and phosphodiesterase 7A (PDE7A), for ELNs-contained cargo. In conclusion, our study indicates that B-ELNs can be considered candidate therapeutic carriers of bioactive compounds potentially able to protect vascular system against various stressors.

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PMID: 

Food Chem Toxicol. 2020 May 6 ;141:111414. Epub 2020 May 6. PMID: 32387444

Abstract Title: 

Chemoprotective effect of carotenoids from Lycium barbarum L. on SH-SY5Y neuroblastoma cells treated with beauvericin.

Abstract: 

Goji berry has recently been introduced in Mediterranean diet and its consumption is increasing. This study aims to determine cytoprotection of lutein (LUT), zeaxanthin (ZEAX) and goji berry extract (GBE) rich in carotenoids against Beauvericin (BEA)-induced cytotoxicity on SH-SY5Y neuroblastoma cells. Both carotenoids and GBE showed cytoprotective effects. Cytoprotection was evaluated by simultaneous combination of the two xanthophylls LUT and ZEAX with BEA, as well as using pre-treatment assays. The highest protective effect occurred in 16%, 24% and 12% respectively for LUT, ZEAX and LUT + ZEAX incubating simultaneously with BEA, while by pre-treatment assay LUT showed a cytoprotection effect over 30% and ZEAX alone or LUT + ZEAX promoted only a slight cytoprotection (

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PMID: 

J Med Food. 2020 May 11. Epub 2020 May 11. PMID: 32392444

Abstract Title: 

Polysaccharide Extracted fromLeaves Ameliorates Asthma in Mice by Reducing Inflammation and Modulating Gut Microbiota.

Abstract: 

This study was designed to explore the impact ofpolysaccharide (LBP) on inflammation and gut microbiota in mice with allergic asthma. Mice were divided into four groups: control group, OVA (ovalbumin) group, Con+LBP group, OVA+LBP group. After 28 days of LBP intervention, mice were euthanized and associated indications were investigated. Histopathological examination demonstrated that LBP reduced lung injury. The results of our current study provide evidence that supplementation with LBP in asthmatic mice decreases,,,, andin plasma and bronchoalveolar lavage fluid (BALF). Sequencing and analysis of gut microbiota indicated that compared with the OVA group,andwere increased, but,,, andwere decreased in the OVA+LBP group. We also found that gut microbiota were related to inflammation-related factors. Therefore, we speculate that LBP may improve allergic asthma by altering gut microbiota and inhibiting inflammation in mice.

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PMID: 

HIV Med. 2020 Apr ;21(4):246-278. Epub 2019 Nov 22. PMID: 31756034

Abstract Title: 

MicroRNAs and exosomes: key players in HIV pathogenesis.

Abstract: 

OBJECTIVES: HIV infection is well known to cause impairment of the human immune system, and until recently was a leading cause of death. It has been shown that T lymphocytes are the main targets of HIV. The virus inactivates T lymphocytes by interfering with a wide range of cellular and molecular targets, leading to suppression of the immune system. The objective of this review is to investigate to what extent microRNAs (miRNAs) are involved in HIV pathogenesis.METHODS: The scientific literature (Pubmed and Google scholar) for the period 1988-2019 was searched.RESULTS: Mounting evidence has revealed that miRNAs are involved in viral replication and immune response, whether by direct targeting of viral transcripts or through indirect modulation of virus-related host pathways. In addition, exosomes have been found to act as nanoscale carriers involved in HIV pathogenesis. These nanovehicles target their cargos (i.e. DNA, RNA, viral proteins and miRNAs) leading to alteration of the behaviour of recipient cells.CONCLUSIONS: miRNAs and exosomes are important players in HIV pathogenesis. Additionally, there are potential diagnostic applications of miRNAs as biomarkers in HIV infection.

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PMID: 

Virulence. 2020 Dec ;11(1):32-38. PMID: 31885311

Abstract Title: 

Exosomes cloak the virion to transmit Enterovirus 71 non-lytically.

Abstract: 

Enterovirus 71 (EV71) is a non-enveloped virus and it can be released from host cells through a traditional cytolytic manner. Now, we showed EV71 could be spread non-lytically between cells during early viral infection. In order to explain this phenomenon, we separated supernatant fluids of rhabdomyosarcoma (RD) cells cultures infected with EV71 by isopycnic gradient centrifugation. Two populations of virus particles were morphology indistinguishable by transmission electron microscope (TEM). It showed that some EV71 particles were wrapped inside extracellular vesicles which were verified to be exosomes by immunoassay and morphologic analysis. In addition, exosomes containing viral RNA were shed in plasma of EV71-infected encephalitis in children. Our findings indicate that the"non-enveloped" EV71 virions could be wrapped within exosomes which promote their spread in the absence of cell lysis.: EV71: enterovirus 71; EXO: exosome; RD: rhabdomyosarcoma; TEM: transmission electron microscope; HFMD: hand, foot, and mouth disease; HIV: immunodeficiency virus; HCV: hepatitis C virus; HTLV: Human T-cell lymphotropic virus; HAV: hepatitis A virus; MOI: multiplicity of infection; EVs: extracellular vesicles; VP1: viral capsid protein 1; NTA: nanoparticle tracking analysis; CNS: central nervous system.

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PMID: 

J Cell Commun Signal. 2020 Feb 14. Epub 2020 Feb 14. PMID: 32060725

Abstract Title: 

Where does the cargo go?: Solutions to provide experimental support for the"extracellular vesicle cargo transfer hypothesis".

Abstract: 

It is widely believed that extracellular vesicles (EVs) mediate intercellular communications by functioning as messengers. EVs contain various biomolecules, including nucleic acids and proteins, as cargo in the internal space. Thus, it has been postulated that this cargo can be transferred from donor cells to recipient cells, leading to phenotypic changes in the recipient cells. However, there is a lack of experimental evidence for the aforementioned hypothesis, that EVs function as messengers. This is presumably because of a lack of rigorous methodologies for EV research. Although cells usually incorporate nanoparticles (NPs) from the extracellular space via endocytosis, these NPs are processed through the endo/lysosomal system and do not escape to the cytoplasm unless they disrupt or fuse with the endo/lysosomal membrane. Whether EVs actually are capable of escaping endo/lysosomes is still debatable. In contrast, viruses have evolved to efficiently deliver their cargo (viral proteins and genetic material) into the cytoplasm of host (recipient) cells by circumventing endo/lysosomal degradation. Thus, it may be helpful to compare EVs to viruses in terms of cargo delivery. The present technological issues that hinder obtaining support for the"EV cargo transfer hypothesis"are summarized and potential solutions for EV research are proposed.

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PMID: 

Curr Clin Microbiol Rep. 2019 Sep ;6(3):121-131. Epub 2019 Jul 3. PMID: 32051811

Abstract Title: 

Extracellular Vesicles in Epstein-Barr Virus Pathogenesis.

Abstract: 

Purpose of review: Epstein-Barr virus (EBV) is a known determinant for numerous malignancies and may contribute to autoimmune diseases. The underlining mechanisms behind EBV pathologies is not completely understood. Recently, extracellular vesicles (EVs) released from infected cells have been found to produce profound effects on cellular microenvironments. Therefore, in this review we sought to critically evaluate the roles of EVs in EBV pathogenesis and assess their potential therapeutic and diagnostic utility.Recent findings: EBV-altered EVs are capable of activating signaling cascades and phenotypic changes in recipient cells through the transfer of viral proteins and RNAs. Moreover, several EV-associated microRNAs have encouraging prognostic or diagnostic potential in EBV-associated cancers.Summary: Current evidence suggests that EBV-modified EVs affect viral pathogenesis and cancer progression. However, further research is needed to investigate the direct role of both viral and host products on recipient cells and the mechanisms driving viral protein and RNA EV packaging and content modification.

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PMID: 

Int J Biol Sci. 2020 ;16(6):904-920. Epub 2020 Jan 22. PMID: 32140061

Abstract Title: 

Exosomes Transmit Viral Genetic Information and Immune Signals may cause Immunosuppression and Immune Tolerance in ALV-J Infected HD11 cells.

Abstract: 

Avian leukosis virus (ALV) is oncogenic retrovirus that not only causes immunosuppression but also enhances the host's susceptibility to secondary infection. Exosomes play vital role in the signal transduction cascades that occur in response to viral infection. We want to explore the function of exosomes in the spread of ALV and the body's subsequent immunological response. RNA-sequencing and the isobaric tags for relative and absolute quantitation (iTRAQ) method were used to detect differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) in exosomes secreted by macrophage cells in response to injection with ALV subgroup J (ALV-J). RNA-sequencing identified 513 DEGs in infected cells, with specific differential regulation in mRNA involved in tight junction signaling, TNF signaling, salmonella infection response, and immune response, among other important cellular processes. Differential regulation was observed in 843 lncRNAs, with particular enrichment in those lncRNA targets involved in Rap1 signaling, HTLV-I infection, tight junction signaling, and other signaling pathways. A total of 50 DEPs were identified in the infected cells by iTRAQ. The proteins enriched are involved in immune response, antigen processing, the formation of both MHC protein and myosin complexes, and transport. Combined analysis of the transcriptome and proteome revealed that there were 337 correlations between RNA and protein enrichment, five of which were significant. Pathways that were enriched on both the RNA and protein levels were involved in pathways in cancer, PI3K-Akt signaling pathway, Endocytosis, Epstein-Barr virus infection. These data show that exosomes are transmitters of intercellular signaling in response to viral infection. Exosomes can carry both viral nucleic acids and proteins, making it possible for exosomes to be involved in the viral infection of other cells and the transmission of immune signals between cells. Our sequencing results confirme previous studies on exosomes and further find exosomes may cause immunosuppression and immune tolerance.

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PMID: 

J Virol. 2020 Apr 15. Epub 2020 Apr 15. PMID: 32295924

Abstract Title: 

hnRNPA2B1 associated with recruitment of RNA into exosomes plays a key role in HSV-1 release from infected cells.

Abstract: 

hnRNPA2B1, an abundant cellular protein has been reported to recruit RNAs bearing a specific sequence (EXO motif) into exosomes. We characterized an exosome population averaging 100+/- 50 nm in diameter and containing a defined set of constitutive exosome markers. This population packages miRNAs and can be directed to block targeted gene expression in a dose dependent fashion. The objectives of these studies were to characterize its role in the recruitment of miRNA. We report 4 key findings (i) hnRNPA2B1 is not a component of exosomes produced in HEp-2 or in HEK293T cells. Hence hnRNPA2B1 carries its cargo at most to the site of exosome assembly but it is not itself incorporated into exosomes. (ii) The accumulation of exosomes produced by cells in which the gene encoding hnRNPA2B1 has been knocked out (ΔhnRNPA2B1) was reduced 3 fold. (iii) In uninfected HEp-2 cells hnRNPA2B1 is localized in the nucleus. In cells infected with herpes simplex virus 1 (HSV-1) hnRNPA2B1 was quantitatively exported to the cytoplasm, at least a fraction of hnRNPA2B1 co-localized with a Golgi marker. Lastly (iv) in ΔhnRNPA2B1 cells there was a 2 to 3-fold reduction in virus yield but a significant (>10 fold) reduction in the HSV-1 released through the apical surface into the extracellular environment. The absence of hnRNPA2B1 had no significant impact on the basolateral export of HSV-1 from infected to uninfected cells by direct cell to cell contact. The results suggest that hnRNPA2B1 plays a key role in the transport of enveloped virus from its site of assembly to the extracellular environment.In this report we show hnRNPA2B1 is not a component of exosomes produced in HEp-2 or in HEK293T cells. In HSV-1 infected cells hnRNPA2B1 was quantitatively translocated from nucleus into the cytoplasm. In infectedΔhnRNPA2B1 cells Golgi dependent transport of virus from the apical surface to the extracellular medium was significantly reduced. In essence, this report supports the hypothesis that hnRNPA2B1 plays a key role in the egress of exosomes and of HSV-1 from infected cells.

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