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Esplanade, OR, United States
Esplanade, OR, United States
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Tumor necrosis factor-α (TNF-α) plays an important role in inflammatory processes. This study examined the effects of natural eggshell membrane (NEM®) (ESM Technologies, LLC, Carthage, MO, USA) on interleukin (IL)-2, IL-4, IL-6, IL-10, interferon-γ (IFN-γ), and TNF-α cytokine production by 4-day peripheral blood mononuclear cell (PBMC) cultures exposed to serial dilutions of either an aqueous extract of natural eggshell membrane (NEM-AQ) or NEM subjected to in vitro digestion (NEM-IVD). The effects on cytokine production were also assessed in the presence of phytohemagglutinin (PHA) and pokeweed mitogen (PWM) where exposure to NEM-AQ resulted in reduced levels of proliferation and statistically significant effects on IL-6, IL-10, IFN-γ, and TNF-α cytokine production. NEM-AQ reduced levels of IL-6, IL-10, IFN-γ, and TNF-α in cultures exposed to PHA. In cultures containing PWM, NEM-AQ reduced production of IL-10 and at the highest dose tested increased IL-6 and decreased TNF-α cytokine levels. NEM-IVD, at the two lowest concentrations of product, significantly reduced TNF-α production by PBMC cultures exposed to PWM compared with the in vitro digest control or native NEM. Taken together, these results suggest that NEM-AQ can influence signaling events in response to the T cell-specific mitogen PHA as well as to the mitogen PWM that require cellular cross-talk and that these effects may be partially mediated through a reduction in level of the pro-inflammatory cytokine TNF-α. The suppression of TNF-α production in the presence of NEM-IVD is promising for the use of NEM as a consumable anti-inflammatory product. © Copyright 2012, Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition.


Kang J.,University of Arkansas for Medical Sciences | Thakali K.M.,University of Arkansas for Medical Sciences | Xie C.,University of Arkansas for Medical Sciences | Kondo M.,Brunswick Laboratories | And 6 more authors.
Food Chemistry | Year: 2012

There are two predominant palm tree species producing edible fruit known as "aaí" found widely dispersed through the Amazon: Euterpe oleracea Mart. and Euterpe precatoria Mart. They differ from each other in terms of how the plants grow and their phytochemical composition. E. oleracea (EO) has received considerable attention as a "super fruit" because of its high antioxidant capacity, while studies on E. precatoria (EP) remain rare. In this study, the antioxidant and anti-inflammatory activities of EP fruit pulps were evaluated by different assays including a series of oxygen radical absorbance capacity (ORAC) based assays, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, the cell-based antioxidant protection in erythrocyte (CAP-e) assay, as well as the nuclear factor-kappa B (NF-κB) secreted embryonic alkaline phosphatase (SEAP) assay. Total phenolics were also measured as an indication of the total phenol content. For comparative purposes, the EO fruit pulp was included. The antioxidant capacity of the EP fruit pulp was determined to be superior to the EO fruit pulp in every chemical based assay. In the cell-based CAP-e assay, the EP fruit pulp showed a dose-dependent inhibition against oxidative damage with an IC 50 of 0.167 g/l. In the SEAP reporter assay, the EP fruit pulp polyphenol-rich extracts inhibited lipopolysaccharide (LPS)-induced NF-κB activation by 23% (p < 0.05) at 20 μg/ml, whereas the extract of the EO fruit pulp did not show a significant inhibitory effect at comparable doses. In addition, carotenoids were quantified for the first time in EP, since EP has high scavenging capacity against singlet oxygen. © 2012 Elsevier Ltd. All rights reserved.


Benson K.F.,NIS Labs | Newman R.A.,University of Houston | Newman R.A.,Nerium Biotechnology | Jensen G.S.,NIS Labs
Clinical, Cosmetic and Investigational Dermatology | Year: 2015

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. © 2015 Benson et al.


Kang J.,University of Arkansas for Medical Sciences | Li Z.,Shanghai Institute of Pharmaceutical Industry | Wu T.,Shanghai Institute of Pharmaceutical Industry | Jensen G.S.,NIS Labs | And 2 more authors.
Food Chemistry | Year: 2010

Acai fruit (Euterpe oleracea Mart.) has been demonstrated to exhibit extremely high anti-oxidant capacity. Seven major flavonoids were isolated from freeze-dried acai pulp by various chromatographic methods. Their structures were elucidated as orientin (1), homoorientin (2), vitexin (3), luteolin (4), chrysoeriol (5), quercetin (6), and dihydrokaempferol (7) by NMR, MS and compared with the reported literature. Compounds 3 and 6 were reported from acai pulp for the first time. Anti-oxidant capacities of these flavonoids were evaluated by oxygen radical absorbance capacity (ORAC) assay, cell-based anti-oxidant protection (CAP-e) assay and reactive oxygen species (ROS) formation in polymorphonuclear (PMN) cells (ROS PMN assay). ORAC values varied distinctly (1420-14,800 μmol TE/g) among the seven compounds based on numbers and positions of hydroxyl groups and/or other substitute groups. The ORAC values of aglycones are generally higher than that of glycosides. CAP-e results indicated that only three compounds (4, 6 and 7) could enter the cytosol and contribute to the reduction of oxidative damage within the cell. The ROS PMN assay showed that five compounds (2-3 and 5-7) demonstrated exceptional effects by reducing ROS formation in PMN cells, which produced high amounts of ROS under oxidative stress. In evaluating the anti-oxidant capacity of natural products, combining both chemical and cell-based assays will provide more comprehensive understanding of anti-oxidant effects and potential biological relevance. © 2010 Elsevier Ltd. All rights reserved.


George A.,Biotropics Malaysia Berhad | Ng C.P.,Cerca Insights Sdn Bhd Technology Development Center Level 2 | O'Callaghan M.,Cerca Insights Sdn Bhd Technology Development Center Level 2 | Jensen G.S.,NIS Labs | Wong H.J.,Biotropics Malaysia Berhad
BMC Complementary and Alternative Medicine | Year: 2014

Background: Polygonum minus Huds.is 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.Method: 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. Results: 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.Conclusion: 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. © 2014 George et al.; licensee BioMed Central Ltd.


Benson K.F.,NIS Labs | Ager D.M.,Cascade Chiropractic and Rehabilitation | Landes B.,Nutritional Products Consulting Group | Aruoma O.I.,Signal Sciences | Jensen G.S.,NIS Labs
Preventive Medicine | Year: 2012

Objective: To evaluate anti-inflammatory properties of a nutraceutical blend containing L-ergothioneine in concert with other anti-inflammatory and analgesic ingredients, combined with nutritional cartilage support. Methodology: Twelve human subjects were tested over a 6-week period of product consumption followed by a 6-week wash-out period, conducted at NIS Labs during late fall/early winter 2010. Range of motion (ROM) assessment of joint motility was performed using JTECH dual digital inclinometry and included flexion, extension, and rotation through the vertical weight-bearing column (neck, thorax, lumbar, hip, knees) and shoulders. Pain evaluation included questionnaires and Visual Analogues Scales regarding primary and secondary pain complaints at rest and at use. Results: ROM improvements were seen after 1. week, and further improved at 6. weeks (primary pain area P< 0.2, secondary pain area P< 0.03). Pain in primary and secondary areas at use was significantly reduced already at 1. week, compared to baseline (P< 0.05). Pain reduction for both primary and secondary pain areas during use reached a high level of statistical significance at 6. weeks (P< 0.004), and remained highly significant after the 6-week wash-out period. Conclusion: Pain reduction and improved ROM were observed during the 6-week consumption. Residual effects were seen 6. weeks after stopping consumption of the ergothioneine supplement. © 2012 Elsevier Inc..


Jensen G.S.,NIS Labs | Patel D.,South Dakota State University | Patel D.,Sterling Technology Inc. | Benson K.F.,NIS Labs
Preventive Medicine | Year: 2012

Objective: To evaluate acute effects of bovine colostrum low-molecular weight fraction (CLMWF) on selected aspects of innate immune function in healthy human subjects. Methodology: A placebo-controlled, double-blinded, randomized cross-over trial involving 12 healthy subjects, age 22-72, was conducted at NIS Labs during the year 2010. Placebo or 150. mg CLMWF was given orally. Blood was drawn immediately before and at 1 and 2. h after consumption. Results: A single dose of CLMWF, when compared to placebo, resulted in rapid increase in phagocytic activity of monocytes at 1h (P<0.12) and polymorphonuclear cells at 1h (P<0.08) and 2h (P<0.03) after consumption. Observations included increased numbers of CD3 + T cells (P<0.05), and a transient reduction in circulating CD3 -CD56 + natural killer (NK) cells at 1h (P<0.04), returning to normal levels at 2h after consumption (P<0.96). The relative increase of NK cells from 1 to 2h after consumption was not associated with an increase in CD69 or CD25 activation markers, suggesting that new NK cells were mobilized into circulation. Conclusion: The increased phagocytic activity and rapid transient changes in NK cell numbers suggest that upon consumption, interaction of CLMWF with immune cells in the gut mucosa triggers immediate events with systemic consequences. © 2012 Elsevier Inc..


Jensen G.S.,NIS Labs | Attridge V.L.,NIS Labs | Lenninger M.R.,NIS Labs | Benson K.F.,NIS Labs
Journal of Medicinal Food | Year: 2015

The goal for this study was to evaluate the effects of daily oral intake of a consumable liquid fermentate containing high-molecular-weight hyaluronan, as well as to perform a basic evaluation of safety and tolerability. A randomized, double-blind placebo-controlled study design was used to examine the effects of oral intake of hyaluronan on chronic pain conditions. Safety assessment included a complete blood count with differential, blood chemistry and electrocardiogram. The study duration was 4 weeks, where three tablespoons (45?mL) product or placebo was ingested during the first 2 weeks, and two tablespoons (30?mL) was consumed during the last 2 weeks. Seventy-eight people between the age of 19 and 71 years enrolled, and 72 people completed the study. Statistical analysis was performed using the two-tailed independent t-test for between-group significance and using the paired t-test for within-group significance. A reduction in pain scores was seen after 2 weeks of consumption of both placebo (P<.1) and active (P<.065) product; the reduction was more pronounced in the group consuming the active test product. Using "within-subject" analysis, a highly significant reduction in chronic pain scores was seen after 2 weeks of consumption of three tablespoons of active product (P<.001), whereas only a mild nonsignificant reduction in pain scores was seen in the placebo group. During the reduced intake for the last 2 weeks of study participation, pain scores showed a slight increase. During the last 2 weeks, a significant increase in the quality of sleep (P<.005) and level of physical energy (P<.05) was seen. The pain reduction during the initial 2 weeks was associated with significant reduction in the use of pain medication (P<.05). Consumption of an oral liquid formula containing high-molecular-weight hyaluronan was associated with relief of chronic pain. © Copyright 2015, Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition 2015.


Benson K.F.,NIS Labs | Beaman J.L.,NIS Labs | Ou B.,Dover science | Okubena A.,Forever Inc. | And 2 more authors.
Journal of Medicinal Food | Year: 2013

The impact of chronic inflammatory conditions on immune function is substantial, and the simultaneous application of anti-inflammatory and immune modulating modalities has potential for reducing inflammation-induced immune suppression. Sorghum-based foods, teas, beers, and extracts are used in traditional medicine, placing an importance on obtaining an increased understanding of the biological effects of sorghum. This study examined selected anti-inflammatory and immune-modulating properties in vitro of Jobelyn™, containing the polyphenol-rich leaf sheaths from a West African variant of Sorghum bicolor (SBLS). Freshly isolated primary human polymorphonuclear (PMN) and mononuclear cell subsets were used to test selected cellular functions in the absence versus presence of aqueous and ethanol extracts of SBLS. Both aqueous and nonaqueous compounds contributed to reduced reactive oxygen species formation by inflammatory PMN cells, and reduced the migration of these cells in response to the inflammatory chemoattractant leukotriene B4. Distinct effects were seen on lymphocyte and monocyte subsets in cultures of peripheral blood mononuclear cells. The aqueous extract of SBLS triggered robust upregulation of the CD69 activation marker on CD3- CD56+ natural killer (NK) cells, whereas the ethanol extract of SBLS triggered similar upregulation of CD69 on CD3+ CD56+ NKT cells, CD3+ T lymphocytes, and monocytes. This was accompanied by many-fold increases in the chemokines RANTES/CCL5, Mip-1α/CCL3, and MIP-1β/CCL4. Both aqueous and nonaqueous compounds contribute to anti-inflammatory effects, combined with multiple effects on immune cell activation status. These observations may help suggest mechanisms of action that contribute to the traditional use of sorghum-based products, beverages, and extracts for immune support. © Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition.


Benson K.F.,NIS Labs | Carter S.G.,NIS Labs | Patterson K.M.,NIS Labs | Patel D.,South Dakota State University | And 2 more authors.
Preventive Medicine | Year: 2012

Objective: To evaluate effects on the innate immune system after exposure to, a consumable low-molecular weight fraction (CLMWF) of immunoglobulin-depleted bovine colostrum whey. Methodology: Cell-based immune assays were performed in vitro, and host resistance towards bacterial and viral infection was evaluated in two mouse studies. Results: In vitro data showed a multimodal effect, as CLMWF treatment resulted in a rapid increase in phagocytosis. CLMWF increased chemotaxis of polymorphonuclear cells towards the bacterial peptide f-MLP. CLMWF treatment of natural killer cells increased expression of the CD69 activation marker. Mononuclear phagocytes showed decreased numbers of CD14 bright and increased number of CD14 dim cells. The remaining CD14 bright cells showed reduced expression of CD80 and CD86, whereas CD14 dim cells showed increased expression of CD80 and CD86, suggesting dendritic cell maturation.Mouse models were applied to evaluate the immune-modulating capacity of CLMWF when consumed acutely during bacterial (Streptococcus) and viral (Influenza) infections in vivo. Reduced bacterial and viral loads were observed in lungs within 24. h. Viral load was also reduced when CLMWF was introduced intranasally. Conclusion: The data suggest that the support of antimicrobial immune defense mechanisms and maturation of antigen-presenting cells in vitro translates to protection in vivo when product is introduced across mucosal membranes. © 2012 Elsevier Inc..

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