Human milk contains large amounts of small extracellular vesicles (sEVs) and their microRNA cargo... more Human milk contains large amounts of small extracellular vesicles (sEVs) and their microRNA cargos, whereas infant formulas contain only trace amounts of sEVs and microRNAs. We assessed the transport of sEVs across the blood-brain barrier (BBB) and sEV accumulation in distinct regions of the brain in brain endothelial cells and suckling mice. We further assessed sEV-dependent gene expression profiles and effects on the dendritic complexity of hippocampal granule cells and phenotypes of EV depletion in neonate, juvenile and adult mice. The transfer of sEVs across the BBB was assessed by using fluorophore-labeled bovine sEVs in brain endothelial bEnd.3 monolayers and dual chamber systems, and in wild-type newborn pups fostered to sEV and cargo tracking (ECT) dams that express sEVs labeled with a CD63-eGFP fusion protein for subsequent analysis by serial two-photon tomography and staining with anti-eGFP antibodies. Effects of EVs on gene expression and dendritic architecture of granule...
Background Milk exosomes and their microRNA (miR) cargos are bioavailable. The content of exosome... more Background Milk exosomes and their microRNA (miR) cargos are bioavailable. The content of exosomes and miRs is negligible in infant formulas compared to human milk, and dietary depletion of exosomes led to changes in bacterial communities and impaired gut health in juvenile mice. Adverse effects of formula feeding may be compounded by using soy formulas due to exosome binding by abundant lectins in that matrix. The purpose of this study was to assess the bioavailability of milk exosomes and their miR cargos added to soy formula in adults, as well as the potential role of soy lectins in exosome bioavailability. Methods Eleven healthy adults (6 men, 5 women) enrolled in this randomized crossover study. Participants consumed 1.0 l of soy formula without (SF) or with (SFE) bovine milk exosomes added. Concentration-time curves of six plasma miRs were analyzed using reverse transcription quantitative PCR. Lectin affinity chromatography was used to assess the binding of exosomes by soy lec...
Additional file 2. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by huma... more Additional file 2. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by human PBMCs ex vivo. PBMCs were isolated before and six hours after milk consumption and treated with or without ConA for 72 h. Boxes represents the 5th and 95th percentiles, horizontal bars are the medians, and error bars indicate the maximum values. *p
Additional file 1. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by huma... more Additional file 1. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by human PBMCs ex vivo. PBMCs were isolated before and six hours after milk consumption and incubated with or without ConA for 48 h. Boxes represents the 5th and 95th percentiles, horizontal bars are the medians, and error bars indicate the maximum values. *p
Background: Human milk contains large amounts of exosomes (MEs) and their regulatory microRNA car... more Background: Human milk contains large amounts of exosomes (MEs) and their regulatory microRNA cargos, whereas infant formulas contain only trace amounts of MEs and microRNAs. Breastfeeding has been implicated in optimal brain development but experimental evidence linking ME intake with brain development is limited. Objectives: We assessed the transport of MEs across the blood-brain barrier (BBB) and ME accumulation in distinct regions of the brain in brain endothelial cells and suckling mice. We further assessed BME-dependent gene expression profiles and effects on the dendritic complexity of hippocampal granule cells and phenotypes of BME depletion in neonate, juvenile and adult mice. Methods: The transfer of MEs across the BBB was assessed by using bovine MEs labeled with FM4-64 or loaded with IRDye-labeled miR-34a in murine brain endothelial bEnd.3 cell monolayers and dual chamber systems, and in wild-type newborn pups fostered to exosome and cargo tracking (ECT) dams that expres...
BackgroundBovine milk exosomes contain cargos such as miRNAs, mRNAs, DNA and proteins. We have de... more BackgroundBovine milk exosomes contain cargos such as miRNAs, mRNAs, DNA and proteins. We have demonstrated that dietary bovine exosomes deliver functional RNA species and regulate genes and metabo...
Background Bovine milk exosomes are studied for their roles as bioactive food compounds and as ve... more Background Bovine milk exosomes are studied for their roles as bioactive food compounds and as vehicles for drug delivery. Both lines of investigation converge on immune function, e.g., immune regulation by absorption of microRNAs encapsulated in milk exosomes across species boundaries, and the possibility of exosomes and their cargos triggering an immune response if used in drug delivery. This study assessed the bioavailability of immune-related microRNAs from bovine milk and changes in plasma cytokine concentrations after milk consumption in humans, and the secretion of cytokines by human peripheral blood mononuclear cells (PBMCs) cultured with milk exosomes transfected with immune-relevant microRNAs. Results Human plasma samples were collected before and at timed intervals after a milk meal and analyzed for concentrations of six immune-relevant microRNAs and nine cytokines. The peak plasma concentrations of miR-15b-5p, miR-21-5p, miR-106b-5p, and miR-223-3p were 60 ± 9.80% to 162...
Objectives Exosomes are natural nanoparticles that facilitate cell-to-cell communication by trans... more Objectives Exosomes are natural nanoparticles that facilitate cell-to-cell communication by transferring RNAs from donor to recipient cells. Previous studies focused on microRNA cargos and disregarded mRNAs. Exosomes and RNA cargos may be absorbed from milk. RNAs elicit antiviral responses by activating interferon type I (IFN) and NF-kappa B (NF-kB) signaling (Figure 1A). To assess whether 1) bovine milk exosomes (BME) contain microbial mRNAs, 2) microbial mRNAs are bioavailable in humans and mice, and 3) microbial mRNAs activate antiviral responses in reporter cells and mice. Methods The mRNA content in BME was assessed by RNA-sequencing. mRNA bioavailability was assessed by RNA-sequencing analysis of human plasma before and 4 h after consumption of 1 L milk and by administering BME transfected with synthetic, fluorophore (IRDye)-labeled Pseudomonas fluorescens capB mRNA to Balb/c mice by oral gavage. (capB mRNA was abundantly expressed in all BME samples sequenced.) Antiviral resp...
Exosomes are natural nanoparticles that play an important role in cell-to-cell communication. Com... more Exosomes are natural nanoparticles that play an important role in cell-to-cell communication. Communication is achieved through the transfer of cargos, such as microRNAs, from donor to recipient cells and binding of exosomes to cell surface receptors. Exosomes and their cargos are also obtained from dietary sources, such as milk. Exosome and cell glycoproteins are crucial for intestinal uptake. A large fraction of milk exosomes accumulates in the brain, whereas the tissue distribution of microRNA cargos varies among distinct species of microRNA. The fraction of milk exosomes that escapes absorption elicits changes in microbial communities in the gut. Dietary depletion of exosomes and their cargos causes a loss of circulating microRNAs and elicits phenotypes such as loss of cognitive performance, increase in purine metabolites, loss of fecundity, and changes in the immune response. Milk exosomes meet the definition of bioactive food compounds. Expected final online publication date f...
Exosomes participate in cell-to-cell communication, facilitated by the transfer of RNAs, proteins... more Exosomes participate in cell-to-cell communication, facilitated by the transfer of RNAs, proteins and lipids from donor to recipient cells. Exosomes and their RNA cargos do not exclusively originate from endogenous synthesis but may also be obtained from dietary sources such as the inter-species transfer of exosomes and RNAs in bovine milk to humans. Here, we assessed the bioavailability and distribution of exosomes and their microRNA cargos from bovine, porcine and murine milk within and across species boundaries. Milk exosomes labeled with fluorophores or fluorescent fusion proteins accumulated in liver, spleen and brain following suckling, oral gavage and intravenous administration in mice and pigs. When synthetic, fluorophore-labeled microRNAs were transfected into bovine milk exosomes and administered to mice, distinct species of microRNAs demonstrated unique distribution profiles and accumulated in intestinal mucosa, spleen, liver, heart or brain. Administration of bovine milk...
Extracellular vesicles (EVs) in milk harbor a variety of compounds, including lipids, proteins, n... more Extracellular vesicles (EVs) in milk harbor a variety of compounds, including lipids, proteins, noncoding RNAs, and mRNAs. Among the various classes of EVs, exosomes are of particular interest, because cargo sorting in exosomes is a regulated, nonrandom process and exosomes play essential roles in cell-to-cell communication. Encapsulation in exosomes confers protection against enzymatic and nonenzymatic degradation of cargos and provides a pathway for cellular uptake of cargos by endocytosis of exosomes. Compelling evidence suggests that exosomes in bovine milk are transported by intestinal cells, vascular endothelial cells, and macrophages in human and rodent cell cultures, and bovine-milk exosomes are delivered to peripheral tissues in mice. Evidence also suggests that cargos in bovine-milk exosomes, in particular RNAs, are delivered to circulating immune cells in humans. Some microRNAs and mRNAs in bovine-milk exosomes may regulate the expression of human genes and be translated ...
Hibiscus sabdariffa L. (sorrel) has been widely used in the development of tropical beverages and... more Hibiscus sabdariffa L. (sorrel) has been widely used in the development of tropical beverages and folk medicine. This study's objective was to investigate the anti-obesity potential of sorrel calyx extracts (methanol and water) on 3T3-L1 adipocyte cells. Phytochemical content, antioxidant potential as DPPH (1, 1-Diphenyl-2-picrylhydrazyl) radical scavenging activity and ferric reducing antioxidant power (FRAP) and enzyme (α-glucosidase, α-amylase, and pancreatic lipase) inhibitory activities were determined in sorrel methanol extracts (SME) and sorrel water extracts (SWE). Effect of SWE and SME on lipid accumulation, lipolysis and apoptosis were tested in 3T3-L1 adipocyte differentiation and maintenance stage of cells at selected concentrations (200-1000 µg/ml) was studied. The total phenolic (GAE mg/100g dry weight) and total flavonoid (mg catechin equi/ 100g dry weight) contents in SME and SWE were 158.31 and 317.27 and 90.77 and 100.08. DPPH% inhibition (IC-50-mg/ml) and FRAP (mmol Fe [II]/100g dry weight) were 0.82 and 0.33 and 1799.13 and 2296.38 for SWE and SME, respectively. SME and SWE inhibited α-glucosidase, α-amylase, and pancreatic lipase activities by more than 40% at 4mg/ml. Significant (p < 0.05) reduction in lipid accumulation and increased glycerol release in 3T3-L1 cells was observed at concentrations ranged from 600 mg/ml of both extracts. Treating cells with SME-1000 µg/ml at differentiation resulted inhibition (p < 0.05) of lipid accumulation by 45% compared to untreated cells. Highest (p < 0.05) (35%) decrease in triglyceride content as well as higher glycerol release was seen in cells exposed to SME at the differentiation stage. Sorrel extracts induced apoptosis in adipocytes at higher concentrations with prominent effect of treating cells at differentiation stage. The results of this study showed effect of sorrel extracts in reduction of lipid accumulation and increase in lipolysis of 3T3-L1 cells.
The purpose of this study was to assess secondary ecosystem succession in the Livingstone Forest ... more The purpose of this study was to assess secondary ecosystem succession in the Livingstone Forest using a combination of remote sensing and ground-based methods. Livingstone Forest is 16 ha research site in the west end of the city of Salisbury in Rowan county, in the North ...
... 5 , Marcus C. Mentzer 5 , Ezra Mutai 4 , Doshie A. Smith 2 , Tucker J. Souther 5 , Rolanda Su... more ... 5 , Marcus C. Mentzer 5 , Ezra Mutai 4 , Doshie A. Smith 2 , Tucker J. Souther 5 , Rolanda Sue 3 , Caleb J. Sutton 2 ... Secondary successional ecosystems are widespread across the southeastern US and, as a whole, comprise a substantial terrestrial sink for atmospheric carbon ...
Human milk contains large amounts of small extracellular vesicles (sEVs) and their microRNA cargo... more Human milk contains large amounts of small extracellular vesicles (sEVs) and their microRNA cargos, whereas infant formulas contain only trace amounts of sEVs and microRNAs. We assessed the transport of sEVs across the blood-brain barrier (BBB) and sEV accumulation in distinct regions of the brain in brain endothelial cells and suckling mice. We further assessed sEV-dependent gene expression profiles and effects on the dendritic complexity of hippocampal granule cells and phenotypes of EV depletion in neonate, juvenile and adult mice. The transfer of sEVs across the BBB was assessed by using fluorophore-labeled bovine sEVs in brain endothelial bEnd.3 monolayers and dual chamber systems, and in wild-type newborn pups fostered to sEV and cargo tracking (ECT) dams that express sEVs labeled with a CD63-eGFP fusion protein for subsequent analysis by serial two-photon tomography and staining with anti-eGFP antibodies. Effects of EVs on gene expression and dendritic architecture of granule...
Background Milk exosomes and their microRNA (miR) cargos are bioavailable. The content of exosome... more Background Milk exosomes and their microRNA (miR) cargos are bioavailable. The content of exosomes and miRs is negligible in infant formulas compared to human milk, and dietary depletion of exosomes led to changes in bacterial communities and impaired gut health in juvenile mice. Adverse effects of formula feeding may be compounded by using soy formulas due to exosome binding by abundant lectins in that matrix. The purpose of this study was to assess the bioavailability of milk exosomes and their miR cargos added to soy formula in adults, as well as the potential role of soy lectins in exosome bioavailability. Methods Eleven healthy adults (6 men, 5 women) enrolled in this randomized crossover study. Participants consumed 1.0 l of soy formula without (SF) or with (SFE) bovine milk exosomes added. Concentration-time curves of six plasma miRs were analyzed using reverse transcription quantitative PCR. Lectin affinity chromatography was used to assess the binding of exosomes by soy lec...
Additional file 2. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by huma... more Additional file 2. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by human PBMCs ex vivo. PBMCs were isolated before and six hours after milk consumption and treated with or without ConA for 72 h. Boxes represents the 5th and 95th percentiles, horizontal bars are the medians, and error bars indicate the maximum values. *p
Additional file 1. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by huma... more Additional file 1. Box plots of TNF-alpha, IL-1β, IL-6, and IL-10 concentrations secreted by human PBMCs ex vivo. PBMCs were isolated before and six hours after milk consumption and incubated with or without ConA for 48 h. Boxes represents the 5th and 95th percentiles, horizontal bars are the medians, and error bars indicate the maximum values. *p
Background: Human milk contains large amounts of exosomes (MEs) and their regulatory microRNA car... more Background: Human milk contains large amounts of exosomes (MEs) and their regulatory microRNA cargos, whereas infant formulas contain only trace amounts of MEs and microRNAs. Breastfeeding has been implicated in optimal brain development but experimental evidence linking ME intake with brain development is limited. Objectives: We assessed the transport of MEs across the blood-brain barrier (BBB) and ME accumulation in distinct regions of the brain in brain endothelial cells and suckling mice. We further assessed BME-dependent gene expression profiles and effects on the dendritic complexity of hippocampal granule cells and phenotypes of BME depletion in neonate, juvenile and adult mice. Methods: The transfer of MEs across the BBB was assessed by using bovine MEs labeled with FM4-64 or loaded with IRDye-labeled miR-34a in murine brain endothelial bEnd.3 cell monolayers and dual chamber systems, and in wild-type newborn pups fostered to exosome and cargo tracking (ECT) dams that expres...
BackgroundBovine milk exosomes contain cargos such as miRNAs, mRNAs, DNA and proteins. We have de... more BackgroundBovine milk exosomes contain cargos such as miRNAs, mRNAs, DNA and proteins. We have demonstrated that dietary bovine exosomes deliver functional RNA species and regulate genes and metabo...
Background Bovine milk exosomes are studied for their roles as bioactive food compounds and as ve... more Background Bovine milk exosomes are studied for their roles as bioactive food compounds and as vehicles for drug delivery. Both lines of investigation converge on immune function, e.g., immune regulation by absorption of microRNAs encapsulated in milk exosomes across species boundaries, and the possibility of exosomes and their cargos triggering an immune response if used in drug delivery. This study assessed the bioavailability of immune-related microRNAs from bovine milk and changes in plasma cytokine concentrations after milk consumption in humans, and the secretion of cytokines by human peripheral blood mononuclear cells (PBMCs) cultured with milk exosomes transfected with immune-relevant microRNAs. Results Human plasma samples were collected before and at timed intervals after a milk meal and analyzed for concentrations of six immune-relevant microRNAs and nine cytokines. The peak plasma concentrations of miR-15b-5p, miR-21-5p, miR-106b-5p, and miR-223-3p were 60 ± 9.80% to 162...
Objectives Exosomes are natural nanoparticles that facilitate cell-to-cell communication by trans... more Objectives Exosomes are natural nanoparticles that facilitate cell-to-cell communication by transferring RNAs from donor to recipient cells. Previous studies focused on microRNA cargos and disregarded mRNAs. Exosomes and RNA cargos may be absorbed from milk. RNAs elicit antiviral responses by activating interferon type I (IFN) and NF-kappa B (NF-kB) signaling (Figure 1A). To assess whether 1) bovine milk exosomes (BME) contain microbial mRNAs, 2) microbial mRNAs are bioavailable in humans and mice, and 3) microbial mRNAs activate antiviral responses in reporter cells and mice. Methods The mRNA content in BME was assessed by RNA-sequencing. mRNA bioavailability was assessed by RNA-sequencing analysis of human plasma before and 4 h after consumption of 1 L milk and by administering BME transfected with synthetic, fluorophore (IRDye)-labeled Pseudomonas fluorescens capB mRNA to Balb/c mice by oral gavage. (capB mRNA was abundantly expressed in all BME samples sequenced.) Antiviral resp...
Exosomes are natural nanoparticles that play an important role in cell-to-cell communication. Com... more Exosomes are natural nanoparticles that play an important role in cell-to-cell communication. Communication is achieved through the transfer of cargos, such as microRNAs, from donor to recipient cells and binding of exosomes to cell surface receptors. Exosomes and their cargos are also obtained from dietary sources, such as milk. Exosome and cell glycoproteins are crucial for intestinal uptake. A large fraction of milk exosomes accumulates in the brain, whereas the tissue distribution of microRNA cargos varies among distinct species of microRNA. The fraction of milk exosomes that escapes absorption elicits changes in microbial communities in the gut. Dietary depletion of exosomes and their cargos causes a loss of circulating microRNAs and elicits phenotypes such as loss of cognitive performance, increase in purine metabolites, loss of fecundity, and changes in the immune response. Milk exosomes meet the definition of bioactive food compounds. Expected final online publication date f...
Exosomes participate in cell-to-cell communication, facilitated by the transfer of RNAs, proteins... more Exosomes participate in cell-to-cell communication, facilitated by the transfer of RNAs, proteins and lipids from donor to recipient cells. Exosomes and their RNA cargos do not exclusively originate from endogenous synthesis but may also be obtained from dietary sources such as the inter-species transfer of exosomes and RNAs in bovine milk to humans. Here, we assessed the bioavailability and distribution of exosomes and their microRNA cargos from bovine, porcine and murine milk within and across species boundaries. Milk exosomes labeled with fluorophores or fluorescent fusion proteins accumulated in liver, spleen and brain following suckling, oral gavage and intravenous administration in mice and pigs. When synthetic, fluorophore-labeled microRNAs were transfected into bovine milk exosomes and administered to mice, distinct species of microRNAs demonstrated unique distribution profiles and accumulated in intestinal mucosa, spleen, liver, heart or brain. Administration of bovine milk...
Extracellular vesicles (EVs) in milk harbor a variety of compounds, including lipids, proteins, n... more Extracellular vesicles (EVs) in milk harbor a variety of compounds, including lipids, proteins, noncoding RNAs, and mRNAs. Among the various classes of EVs, exosomes are of particular interest, because cargo sorting in exosomes is a regulated, nonrandom process and exosomes play essential roles in cell-to-cell communication. Encapsulation in exosomes confers protection against enzymatic and nonenzymatic degradation of cargos and provides a pathway for cellular uptake of cargos by endocytosis of exosomes. Compelling evidence suggests that exosomes in bovine milk are transported by intestinal cells, vascular endothelial cells, and macrophages in human and rodent cell cultures, and bovine-milk exosomes are delivered to peripheral tissues in mice. Evidence also suggests that cargos in bovine-milk exosomes, in particular RNAs, are delivered to circulating immune cells in humans. Some microRNAs and mRNAs in bovine-milk exosomes may regulate the expression of human genes and be translated ...
Hibiscus sabdariffa L. (sorrel) has been widely used in the development of tropical beverages and... more Hibiscus sabdariffa L. (sorrel) has been widely used in the development of tropical beverages and folk medicine. This study's objective was to investigate the anti-obesity potential of sorrel calyx extracts (methanol and water) on 3T3-L1 adipocyte cells. Phytochemical content, antioxidant potential as DPPH (1, 1-Diphenyl-2-picrylhydrazyl) radical scavenging activity and ferric reducing antioxidant power (FRAP) and enzyme (α-glucosidase, α-amylase, and pancreatic lipase) inhibitory activities were determined in sorrel methanol extracts (SME) and sorrel water extracts (SWE). Effect of SWE and SME on lipid accumulation, lipolysis and apoptosis were tested in 3T3-L1 adipocyte differentiation and maintenance stage of cells at selected concentrations (200-1000 µg/ml) was studied. The total phenolic (GAE mg/100g dry weight) and total flavonoid (mg catechin equi/ 100g dry weight) contents in SME and SWE were 158.31 and 317.27 and 90.77 and 100.08. DPPH% inhibition (IC-50-mg/ml) and FRAP (mmol Fe [II]/100g dry weight) were 0.82 and 0.33 and 1799.13 and 2296.38 for SWE and SME, respectively. SME and SWE inhibited α-glucosidase, α-amylase, and pancreatic lipase activities by more than 40% at 4mg/ml. Significant (p < 0.05) reduction in lipid accumulation and increased glycerol release in 3T3-L1 cells was observed at concentrations ranged from 600 mg/ml of both extracts. Treating cells with SME-1000 µg/ml at differentiation resulted inhibition (p < 0.05) of lipid accumulation by 45% compared to untreated cells. Highest (p < 0.05) (35%) decrease in triglyceride content as well as higher glycerol release was seen in cells exposed to SME at the differentiation stage. Sorrel extracts induced apoptosis in adipocytes at higher concentrations with prominent effect of treating cells at differentiation stage. The results of this study showed effect of sorrel extracts in reduction of lipid accumulation and increase in lipolysis of 3T3-L1 cells.
The purpose of this study was to assess secondary ecosystem succession in the Livingstone Forest ... more The purpose of this study was to assess secondary ecosystem succession in the Livingstone Forest using a combination of remote sensing and ground-based methods. Livingstone Forest is 16 ha research site in the west end of the city of Salisbury in Rowan county, in the North ...
... 5 , Marcus C. Mentzer 5 , Ezra Mutai 4 , Doshie A. Smith 2 , Tucker J. Souther 5 , Rolanda Su... more ... 5 , Marcus C. Mentzer 5 , Ezra Mutai 4 , Doshie A. Smith 2 , Tucker J. Souther 5 , Rolanda Sue 3 , Caleb J. Sutton 2 ... Secondary successional ecosystems are widespread across the southeastern US and, as a whole, comprise a substantial terrestrial sink for atmospheric carbon ...
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