Project Type: Peer Reviewed Publications


Anti-Inflammatory Properties of a Dried Fermentate In Vitro and In Vivo

ABSTRACT

The aim of this study was to document anti-inflammatory properties of a dried fermentate derived from Saccharomyces cerevisiae (EpiCor®), hereafter referred to as dried fermentate in vitro using cell-based bioassays, and in vivo using a skin irritation model in healthy humans. In vitro testing involved parallel assessment of primary human polymorphonuclear (PMN) cell formation of reactive oxygen species (ROS) and migration toward the inflammatory mediator Leukotriene B4. In vivo evaluation used a single-blind placebo-controlled design, where dermal histamine-induced inflammation was used as a model for the complex intercellular signals involved in the initiation, escalation, and resolution of the inflammatory response. Microvascular blood perfusion was evaluated using noninvasive laser Doppler probes applied to the inner forearms of 12 healthy human subjects, where parallel sites were treated with either dried fermentate or saline (placebo). Subjective scores of dermal irritation were also collected. Treatment of PMN cells in vitro resulted in reduced ROS formation and migratory activity toward Leukotriene B4. Clinical results demonstrated significantly reduced microvascular inflammatory responses to histamine-induced skin inflammation, and significantly reduced subjective scores of irritation at the inflamed sites treated with dried fermentate compared with the sites treated with placebo (P<.05). Treatment of inflammatory cells in vitro with dried fermentate resulted in reduced inflammatory responses. This was confirmed in vivo, suggesting that the dried fermentate facilitates the resolution of inflammatory responses. The effects using a topical skin model suggest that similar events may happen when the dried fermentate is introduced across mucosal membranes after consumption.

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Oral intake of a liquid high-molecular-weight hyaluronan associated with relief of chronic pain and reduced use of pain medication: results of a randomized, placebo-controlled double-blind pilot study.

Abstract

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.

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

Abstract

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.

Abstract

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.

Abstract

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.

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Clinical efficacy of a West African sorghum bicolor-based traditional herbal preparation Jobelyn shows increased hemoglobin and CD4+ T-lymphocyte counts in HIV-positive patients.

Abstract

OBJECTIVES:

The purpose of this study was to evaluate a traditional herbal preparation, Jobelyn,® for its effects on anemia and CD4+ T-cell counts in human immunodeficiency virus-positive (HIV+) patients in Nigeria.

DESIGN:

An open-label pilot study involving 10 confirmed (HIV+) patients who were not receiving antiretroviral therapy (ARVT) was performed, in which the patients consumed Jobelyn for 8 weeks, at a dose of 500 mg twice daily. The pilot study was followed by a controlled trial involving 51 patients, all confirmed HIV+, where the patients with CD4+ T-cell counts below 350 cells/μL were receiving ARVT. The eight patients with baseline CD4+ T-cell counts above 350 cells/μL received Jobelyn. The remaining patients who all received ARVT were randomized to ARVT alone versus ARVT+Jobelyn for 12 weeks.

RESULTS:

Patients receiving ARVT showed a statistically significant improvement in their CD4+ T-cell counts across the 12-week study period (p<0.01). Patients receiving ARVT+Jobelyn showed a faster improvement, reaching a high level of statistical significance compared to baseline already at 6 weeks (p<0.001), and remained highly significant at 12 weeks (p<0.001).

CONCLUSIONS:

This is the first controlled study conducted to evaluate efficacy of Jobelyn on immune status in HIV+ patients. The data suggest that consumption of Jobelyn contributed to improved hemoglobin levels and increased CD4+ T-cell counts in Nigerian HIV+ patients. Further studies are needed to examine similar effects in other populations, and to elaborate on the underlying mechanisms, specifically, whether the consumption of Jobelyn supported multiple aspects of bone marrow function.

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West African Sorghum bicolor Leaf Sheaths Have Anti-Inflammatory and Immune-Modulating Properties In Vitro

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.

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Bioactivities of açaí (Euterpe precatoria Mart.) fruit pulp, superior antioxidant and anti-inflammatory properties to Euterpe oleracea Mart.

There are two predominant palm tree species producing edible fruit known as “açaí” 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 IC50of 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.


A novel extract from bovine colostrum whey supports innate immune functions. II. Rapid changes in cellular immune function in humans Preventive Medicine.

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 2h 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.

 

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Improvement of joint range of motion (ROM) and reduction of chronic pain after consumption of an ergothioneine-containing nutritional supplement.

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.

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