Topographical changes in high-protein, milk powders as a function of moisture sorption using amplitude-modulation atomic force microscopy

Mishra, Vinay; Ochalski, Tomasz J.; McCarthy, Noel A.; Brodkorb, André; Rodriguez, Brian J.; Hogan, Sean A.

doi:10.1016/j.foodhyd.2022.107504

Cited by 5 publications

(2 citation statements)

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“…It is known that EVs can withstand a range of pH (2)(3)(4)(5)(6)(7)(8)(9)(10)(11), with pH 4.6 routinely used to remove caseins before collection of high-quality EVs. [5,21,31] In addition, the degree of lipolysis is low in the gas-tric phase in adults and infants compared to the intestinal phase, suggesting that gastric lipase has limited effects on EVs. [25] Salivary EVs have been digested with 0.03-3 mg mL −1 pepsin for 3 h at pH 3 with no difference in their morphology by transmission electron microscopy, but pepsin activity was not reported.…”

Section: Discussionmentioning

confidence: 99%

“…[30] The spring constant of the tip ranged between 32.2 and 35.2 N m −1 . The topographical studies were performed in air using amplitude-modulated intermittent contact mode as described by Mishra et al [31] The surface was scanned at 10 and 1.5 μm 2 in 512 × 512 pixels to visualize EVs cluster morphology and obtain close-ups of single vesicles. Image acquisition and data analyses were performed using Asylum Research software (Ver.…”

Section: Atomic Force Microscopymentioning

confidence: 99%

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Effect of In Vitro Enzyme Digestion and Bile Treatment on Milk Extracellular Vesicles Stability

Oliver,

Mishra,

Santoro

et al. 2024

Molecular Nutrition Food Res

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ScopeMilk extracellular vesicles (EVs) are nanosized particles with potential immune bioactivities. This study examines their fate during in vitro infant gastrointestinal digestion (GI).Methods and resultsBovine milk is digested using the in vitro INFOGEST method, adjusted for the infant. To unravel the contribution of digestive enzymes from bile, milk is treated with digestive enzymes, bile, or a combination of both. EVs are collected posttreatment using differential ultracentrifugation. EVs characterization includes electrophoresis, immunoblotting, nanoparticle tracking analysis, and atomic force microscopy. EVs protein markers programmed cell death 6‐interacting protein (ALIX), tumor susceptibility gene 101 (TSG101), cluster of differentiation 9 (CD9), and xanthine dehydrogenase (XDH) are detected after gastric digestion (G60), but their signal intensity is significantly reduced by intestinal conditions (p < 0.05). Enzyme digestion, compared to bile treatment (I60 + bile), results in a significant reduction of signal intensities for TSG101 and CD9 (p < 0.05). Nanoparticle tracking analysis shows a significant reduction (p < 0.05) of EV numbers at the end of the intestinal phase. EVs are detected by atomic force microscopy at the end of the intestinal phase, showing that intact EVs can survive upper gut digestion.ConclusionIntact EVs can be found at the end of the intestinal phase. However, digestive enzymes and bile reduce the quantity and characteristics of EVs, with digestive enzymes playing a larger role.

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