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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV250074	2/2	Prunus amygdalus var. dulcis	homogenates	DC	Santangelo, Carmen	2024	38%
Study summary Full title Biophysical features of plant-derived nanovesicles: Focus on almonds All authors Carmen Santangelo, Enrico Binetti, S.N. Afifa Azman, Danilo Bondi, Virgilio Brunetti, Marco Farina, Cristina Purcaro, Lorenzo Marramiero, Rossella Di Raimo, Giulia Pietrangelo, Ester Sara Di Filippo, Tiziana Pietrangelo Journal Journal of Food Composition and Analysis Abstract Almond is a traditional food with established beneficial effects on health. Nothing is known about t (show more...)Almond is a traditional food with established beneficial effects on health. Nothing is known about the presence of extracellular vesicles (EVs), recently isolated from other plant material, ingested with food, or as engineered bioactive nanovectors. Aiming to develop and optimize a method to isolate almonds derived nano vesicles (ADNVs), we tested different protocols on pure, blanched and roasted almonds, and investigated the resulting biophysical features. The most feasible and valid method was the sucrose-based ultracentrifugation (DGUC). Size distribution ranged on average 100–200 nm. A negative zeta-potential in the range of –27 to –21 mV has been measured. Microscopy showed a spheroid morphology and preserved structural integrity of isolated ADNVs. Pure almond ADNVs showed the greater amount of TET8; PEN1 was also found, although cooking treatments resulted in a decrease. Attention should be paid to sample managing during DGUC steps and to sample deposition and drying for microscopy procedures. Our findings contribute to enriching the hot issue on EVs research from edible sources, giving reason to their possible role in interspecies communication, and their exploitation for the delivery of bioactive compounds or therapeutics molecules. The potentiality of almonds' industrial residuals should be investigated in further research. (hide) EV-METRIC 38% (75th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type homogenates Sample origin Cooked Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture Density cushion Protein markers EV: TET8/ PEN1 non-EV: None Proteomics no Show all info Study aim biophysical features Sample Species Prunus amygdalus var. dulcis Sample Type homogenates Separation Method Density cushion Density medium Sucrose Sample volume 8 Cushion volume 1 Density of the cushion 30% Centrifugation time 90 Centrifugation speed 100000 Characterization: Protein analysis Protein Concentration Method UV absorbance Western Blot Detected EV-associated proteins TET8/ PEN1 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Mean Reported size (nm) 138.9 EV concentration Yes Particle yield particles per milliliter of starting sample: 5.47e11 EM EM-type Atomic force microscopy Image type Close-up, Wide-field Report size (nm) 142

	EV250074	1/2	Prunus amygdalus var. dulcis	homogenates	DC	Santangelo, Carmen	2024	25%
Study summary Full title Biophysical features of plant-derived nanovesicles: Focus on almonds All authors Carmen Santangelo, Enrico Binetti, S.N. Afifa Azman, Danilo Bondi, Virgilio Brunetti, Marco Farina, Cristina Purcaro, Lorenzo Marramiero, Rossella Di Raimo, Giulia Pietrangelo, Ester Sara Di Filippo, Tiziana Pietrangelo Journal Journal of Food Composition and Analysis Abstract Almond is a traditional food with established beneficial effects on health. Nothing is known about t (show more...)Almond is a traditional food with established beneficial effects on health. Nothing is known about the presence of extracellular vesicles (EVs), recently isolated from other plant material, ingested with food, or as engineered bioactive nanovectors. Aiming to develop and optimize a method to isolate almonds derived nano vesicles (ADNVs), we tested different protocols on pure, blanched and roasted almonds, and investigated the resulting biophysical features. The most feasible and valid method was the sucrose-based ultracentrifugation (DGUC). Size distribution ranged on average 100–200 nm. A negative zeta-potential in the range of –27 to –21 mV has been measured. Microscopy showed a spheroid morphology and preserved structural integrity of isolated ADNVs. Pure almond ADNVs showed the greater amount of TET8; PEN1 was also found, although cooking treatments resulted in a decrease. Attention should be paid to sample managing during DGUC steps and to sample deposition and drying for microscopy procedures. Our findings contribute to enriching the hot issue on EVs research from edible sources, giving reason to their possible role in interspecies communication, and their exploitation for the delivery of bioactive compounds or therapeutics molecules. The potentiality of almonds' industrial residuals should be investigated in further research. (hide) EV-METRIC 25% (25th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type homogenates Sample origin Control condition Focus vesicles extracellular vesicle Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture Density cushion Protein markers EV: TET8/ PEN1 non-EV: None Proteomics no Show all info Study aim biophysical features Sample Species Prunus amygdalus var. dulcis Sample Type homogenates Separation Method Density cushion Density medium Sucrose Sample volume 8 Cushion volume 1 Density of the cushion 30% Centrifugation time 90 Centrifugation speed 100000 Characterization: Protein analysis Protein Concentration Method UV absorbance Western Blot Detected EV-associated proteins TET8/ PEN1 Characterization: Lipid analysis No Characterization: Particle analysis DLS Report type Mean Reported size (nm) 174.3 NTA Report type Mean Reported size (nm) 130.8 EV concentration Yes Particle yield particles per milliliter of starting sample: 7.73e11 EM EM-type Transmission-EM Image type Close-up

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EV-TRACK ID	EV250074
species	Prunus amygdalus var. dulcis
sample type	homogenates
condition	Cooked	Control condition
separation protocol	DC	DC
Exp. nr.	2	1
EV-METRIC %	38	25