Project Tag: CAP-e

Antioxidant and Immune-Modulatory Effects of Aloe vera Decolorized Leaf Juice vs. Inner Leaf Juice

The aloe vera (Aloe vera) plant has a long history of traditional therapeutic topical use. Its leaf and inner leaf juices are also ingested in various forms. Aloe vera leaf juice is made from macerated aloe leaves, and when it is intended for human ingestion, it is purified of latex constituents via a charcoal filtration process known as decolorization. Several publications have shown immunomodulatory effects, as well as antibacterial, antifungal and anti-parasitic properties of aloe vera inner leaf juice, but limited information is available regarding these effects for aloe vera leaf juice after the decolorization process. The aim of this preliminary study was to compare biologically-relevant antioxidant and immune-modulatory effects of aloe vera inner leaf and decolorized leaf preparations using a variety of in vitro techniques.

ExpBio poster 5-1-17


Antioxidant, anti-inflammatory, anti-apoptotic, and skin regenerative properties of an Aloe vera-based extract of Nerium oleander leaves (NAE-8®)


Objective: The goal for this study was to evaluate the effects of an Aloe vera-based Nerium oleander extract (NAE-8®), compared to an extract of A. vera gel alone (ALOE), and to an aqueous extract of N. oleander (AQ-NOE) in bioassays pertaining to dermatologic potential with respect to antioxidant protection, anti-inflammatory effects, and cytokine profiles in vitro.
Methods: Cellular antioxidant protection was evaluated in three separate bioassays: The cellular antioxidant protection of erythrocytes (CAP-e) assay, protection of cellular viability and prevention of apoptosis, and protection of intracellular reduced glutathione levels, where the last two assays were performed using human primary dermal fibroblasts. Reduction of intracellular formation of reactive oxygen species (ROS) was tested using polymorphonuclear cells in the absence and presence of oxidative stress. Changes to cytokine and chemokine profiles when whole blood cells and human primary dermal fibroblasts were exposed to test products were determined using a 40-plex Luminex array as a method for exploring the potential cross-talk between circulating and skin-resident cells.
Results: The NAE-8® provided significantly better antioxidant protection in the CAP-e bioassay than AQ-NOE. NAE-8® and AQ-NOE both protected cellular viability and intracellular reduced glutathione, and reduced the ROS formation significantly when compared to control cells, both under inflamed and neutral culture conditions. ALOE showed minimal effect in these bioassays. In contrast to the NAE-8®, the AQ-NOE showed induction of inflammation in the whole blood cultures, as evidenced by the high induction of CD69 expression and secretion of a number of inflammatory cytokines. The treatment of dermal fibroblasts with NAE-8® resulted in selective secretion of cytokines involved in collagen and hyaluronan production as well as re-epithelialization during wound healing.
Conclusion: NAE-8®, a novel component of a commercial cosmetic product, showed beneficial antioxidant protection in several cellular models, without the induction of leukocyte activation and secretion of inflammatory cytokines. The biological efficacy of NAE-8® was unique from both ALOE and AQ-NOE.

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Antioxidant and Anti-Inflammatory Properties of an Aqueous Cyanophyta Extract Derived from Arthrospira Platensis: Contribution to Bioactivities by the Non-Phycocyanin Aqueous Fraction.


The goal for this work was to characterize basic biological properties of a novel Arthrospira platensis-based aqueous cyanophyta extract (ACE), enriched in the known anti-inflammatory cyclooxygenase-2 (COX-2) inhibitor phycocyanin (PC), but also containing a high level of non-PC bioactive compounds. Antioxidant properties were tested in parallel in the Folin-Ciocalteu assay (chemical antioxidant capacity) and in the cellular antioxidantprotection (CAP-e) bioassay, where both the PC and the non-PC fractions contributed to the antioxidant capacity and CAP of ACE. In contrast to the COX-2 inhibition seen in the presence of PC, the inhibition of enzymatic activity of the inflammatory mediator Lipoxygenase was associated specifically with the non-PC fraction of ACE. Inhibition of formation of reactive oxygen species (ROS) was evaluated using polymorphonuclear cells from healthy human donors. The inhibition of ROS formation was seen for both the PC and non-PC fractions, with ACE showing the most robust effect. The effects of PC, non-PC, and ACE on clotting and clot lysing was tested using a modified Euglobulin fibrinolytic assay in vitro. In the presence of PC, non-PC, and ACE, the time for clot formation and lysis was not affected; however, the clots were significantly more robust. This effect was statistically significant (p<.05) at doses between 125-500 μg/mL, and returned to baseline at lower doses. Both PC and the non-PC fraction contributed to the antioxidant properties and anti-inflammatory effects, without a negative impact on blood clotting in vitro. This suggests a potential benefit for the consumable ACE extract in assisting the reduction of inflammatory conditions.

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Phenolic acids of the two major blueberry species in the US market and their antioxidant and anti-inflammatory activities.


Highbush (cultivated) and lowbush (wild) are the two major blueberry species in the US market. Eight phenolic acids were detected and quantified from these two species by HPLC-MS. Chlorogenic acid was found to be the predominant phenolic acid in both species, with 0.44 mg/g fresh weight in lowbush blueberries and 0.13 mg/g fresh weight in highbush blueberries. Total phenolic content in lowbush blueberries is over three times higher than that of highbush blueberries. The phenolic acid mixtures representing those in the two species were prepared by using authentic standards to assess their contribution to total antioxidant and anti-inflammatory activities of the whole berries. Neither lowbush nor highbush blueberry phenolic acid mixture contributed significantly to the total antioxidant capacity of their relevant whole berries measured by oxygen radical absorbance capacity (ORAC). Both phenolic acid mixtures were able to enter the cell and showed in cell antioxidant activities from the cell based antioxidant protection of erythrocytes (CAP-e) assay. Lowbush blueberry phenolic acid mixture was found to show anti-inflammatory activities by inhibiting the nuclear factor-κB (NF-κB) activation and the production of inflammatory cytokines (TNF-α and IL-6) at the high dose.

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Consumption of dried apple peel powder increases joint function and range of motion.


The goal for this study was to evaluate the effects of consumption of dried apple peel powder (DAPP) on joint function and range of motion (ROM). Additional in vitro and clinical testing was performed to suggest specific mechanisms of action. An open-label clinical pilot study involved 12 healthy people with moderate loss of joint ROM and associated chronic pain. The subjects consumed 4.25 g DAPP daily for 12 weeks, with evaluations at baseline, 2, 4, 8, and 12 weeks. ROM was evaluated at each visit using dual digital inclinometry. Pain scores were collected using Visual Analogue Scales. Blood draws enabled testing of serum antioxidant protective capacity using the cellular antioxidant protection (CAP-e) bioassay. Additional in vitro testing involved testing of cyclooxygenase-2 (COX-2) and lipoxygenase inhibition, cellular antioxidant protection by the CAP-e bioassay, and formation of reactive oxygen species (ROS) by polymorphonuclear (PMN) cells by flow cytometry. Twelve weeks of consumption of DAPP was associated with improved ROM. DAPP provided antioxidants that were available to enter into and protect cells from oxidative damage in vitro, and consumption of DAPP for 12 weeks was associated with a statistically significant improvement in serum antioxidant protective status. DAPP inhibited both COX-2 and lipoxygenase enzymes, and pretreatment of inflammatory PMN cells with DAPP before inflammatory stimulus resulted in reduced ROS formation. This suggests multifaceted anti-inflammatory properties of DAPP. Consumption of DAPP was associated with improved joint function and improved serum antioxidant protection status. The observed pain reduction may be associated with the improved antioxidant status and linked to the apple polyphenols’ anti-inflammatory effects.

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In vitro and ex-vivo cellular antioxidant protection and cognitive enhancing effects of an extract of Polygonum minus Huds (Lineminus™) demonstrated in a Barnes Maze animal model for memory and learning.



Polygonum minus a culinary flavouring that is common in South East Asian cuisine and as a remedy for diverse maladies ranging from indigestion to poor eyesight. The leaves of this herb have been reported to be high in antioxidants. Flavonoids which have been associated with memory, cognition and protection against neurodegeneration were found in P. minus.


This study examined a P. minus aqueous extract (Lineminus™) for its antioxidant activity using the Oxygen Radical Absorbance Capacity (ORAC) assay, the ex vivo Cellular Antioxidant Protection of erythrocytes (CAP-e) assays and for potential anticholinesterase activity in vitro. Cognitive function and learning of Lineminus™ was evaluated using scopolamine induced cognition deficits in a Barnes maze, rodent model of cognition.


The extract displayed in vitro antioxidant activity with a total ORAC value of 16,964 μmole TE/gram. Cellular antioxidant protection from free radical damage using the CAP-e assay, with an IC50 of 0.58 g/L for inhibition of cellular oxidative damage, was observed. The extract inhibited cholinesterase activity with an IC50 of 0.04 mg/ml with a maximum inhibition of 68%. In a rodent model of cognition using scopolamine induced cognition deficits in the Barnes maze, the extract attenuated scopolamine induced disruptions in learning at the higher dose of 100 mg/kg.


These data shows that P. minus possesses antioxidant and anticholinesterase activity and demonstrated enhanced cognition in vivo. The data suggest neuroprotective properties of the extract.

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Analgesic and anti-inflammatory properties of a phycocyanin-enriched microalgae extract CyaninPlus.

Objective and methodology

The objective for this research was to document the anti-inflammatory  properties Of the microalgae extract in vitro and in vivo.

A panel of in vitro tests addressed the following:
  • Antioxidant capacity
  • Inhibition of inflammatory enzymes COX-2 and Lipoxygenase
  • Reduction of free radical production by inflammatory cells
Two human clinical pilot studies were performed:
  • An open-label 4-week study on 12 people 40-70 years of age suffering from chronic joint pain
  • A double-blinded placebo-controlled cross-over study involving 12 people in the same age range, where each consumption phase was 1 week, separated by a wash-out period of one week.

Poster (pdf)

Antioxidant bioavailability and rapid immune-modulating effects after consumption of a single acute dose of a high-metabolite yeast immunogen: results of a placebo-controlled double-blinded crossover pilot study.

The objective of this pilot study was to investigate the acute effects on circulating lymphocyte subsets, antioxidant status, and cytokine profile after consumption of EpiCor(®) (EP) (Embria Health Sciences, Ankeny, IA, USA), a dried fermentate produced from Saccharomyces cerevisiae, using a placebo-controlled randomized crossover study design with 12 healthy adult human subjects. EP contains high levels of bioavailable antioxidants and strongly activates natural killer (NK) cells in vitro. EP consumption has been shown to increase erythrocyte hematocrit levels, boost mucosal immune protection, reduce cold/flu symptoms, reduce seasonal allergy symptoms and the need for rescue medication, and increase salivary secretory immunoglobulin A levels. This warranted further study on immune effects in humans. A within-subject analysis of data collected before and at 1 and 2 hours after consumption of a single dose of 500 mg of EP versus placebo was performed. A transient reduction in circulating T and NK cell numbers was observed 2 hours post-consumption, suggesting that homing and recirculation of these cells, as part of healthy immune surveillance, were supported by EP. The increased expression of activation markers on the CD3(-) CD56(+) NK cell population was significant for CD69 at 1 hour post-consumption (CD25, P<.07; CD69, P<.05), whereas for CD25 it was significant at 2 hours after consumption (CD25, P<.03; CD69, P<.15). A rapid increase in serum interferon-γ was observed at 1 hour post-consumption (P<.07; after removal of two outlying data sets, P<.05) and may have contributed to the effects seen on NK and T cell subsets. Significant increase in serum antioxidant protection was seen 2 hours after consumption (P<.04). Thus consumption of a single 500 mg dose of EP provides a rapid and transient effect on the trafficking and activation status of specific lymphocyte subsets, as well as increased antioxidant protection.

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Evaluation of bioavailability of antioxidants in EpiCor at the cellular level and in clinical studies, using the cell-based antioxidant protection of erythrocytes (CAP-e) bioassay

Purpose of the study

The objective of this study was to evaluate EpiCor» antioxidant bioavailability.  One aspect of bioavailability is to examine whether compounds are capable of crossing the cell membrane and entering live cells. Performing cell-based testing for antioxidant protection involves the dynamics Of living cells, their enzymes, and membrane properties.

An improvement in serum antioxidant protection capacity after ingestion of a food or natural product may reflect the content of easily absorbed antioxidants existing in the native product, as well as antioxidant compounds released or generated as a result of normal digestive processes. Unknown metabolites may contribute significantly to the in vivo antioxidant protection. By using the CAP-e assay to evaluate the serum capacity for antioxidant protection, such metabolites do not need to be chemically identified; as long as they are able to enter and protect living cells from oxidative damage they contribute to the CAP-e results. This is in contrast to other types of bioavailability studies, where chemical analysis is performed on serum to examine the sample for metabolites known to have come from the digestion of a natural product.

Poster (pdf)

Açai (Euterpe oleracea) An Amazonian Palm Fruit with Broad Antioxidant and Anti-Inflammatory Activities

The pulp of the Amazonian palm fruit, açai (Euterpe oleracea Mart.), has been found to have exceptional antioxidant capacity in vitro, particularly against the superoxide, peroxyl, and hydroxyl radicals. Studying foods for their potential health benefits to humans can benefit fromin vitro assays that can correlate with in vivo outcomes. Using the cell-based protection of erthrocytes (CAP-e) assay can determine if red blood cells exposed to foods allow antioxidants to enter into cells and protect cells from oxidative damage. Using this assay, the reduction in fluoresecence has been shown to be proportional to the level of intracellular antioxidant protection. In comparing results from blood samples taken during a randomized, double-blind, placebo-controlled, cross over study involving an açai-based fruit juice taken by subjects in a state of oxidative stress, the CAP-e assay correlated well with levels of antioxidant compounds in cells and reduction in lipid peroxidation. A dose dependent reduction in formation of reactive oxygen species in polymorphonuclear (PMN) cells has shown açai pulp to be active even when diluted down to 0.1 picogram/mL, suggestive of cellular signaling, and indicative that the pulp may contain compounds with the potential to be a food able to exhibit potent anti-inflammatory activity in vivo. A pilot study on range of motion and joint pain in older adults supports this hypothesis, when studied in in older subjects with mild to moderate inflammation of the knees and lumbar spine.

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