This document compares three methods for evaluating the antioxidant potential of foods and natural products: the oxygen radical absorbance capacity (ORAC) assay, a cell-based antioxidant protection assay using erythrocytes (CAP-e), and an assay measuring reactive oxygen species formation in polymorphonuclear cells (ROS PMN). The document applies these three methods to four natural products and finds that while the products showed similar antioxidant properties by ORAC, they had different effects on human cells in the CAP-e and ROS PMN assays. Specifically, Acai provided strong inhibition of ROS formation, indicating anti-inflammatory effects, while Immunel and EpiCor mildly enhanced ROS formation, suggesting activation of the innate immune response. This illustrates that complex
LABORATORY ANALYSIS OF ANTIOXIDANTS IN FOODNutraLima
To characterize the antioxidants in food, it is necessary to know:
-How much antioxidant is contained in the food.
-Are the antioxidants absorbed from the food to the blood?
-If the antioxidants in the blood make their way to the intended body cells.
-Do the antioxidants help the cells to defend themselves against oxygen radicals? May it be given by increasing the production of the cell's own defense enzymes or by direct chemical neutralization of the radicals?
For all these above-mentioned steps different measurement techniques exist.
LABORATORY ANALYSIS OF ANTIOXIDANTS IN FOODNutraLima
To characterize the antioxidants in food, it is necessary to know:
-How much antioxidant is contained in the food.
-Are the antioxidants absorbed from the food to the blood?
-If the antioxidants in the blood make their way to the intended body cells.
-Do the antioxidants help the cells to defend themselves against oxygen radicals? May it be given by increasing the production of the cell's own defense enzymes or by direct chemical neutralization of the radicals?
For all these above-mentioned steps different measurement techniques exist.
Reactive oxygen and nitrogen species (RONS) are involved in deleterious/beneficial biological processes.
The present study sought to investigate the capacity of single and combinatorial herbal formulations of
Acanthus montanus, Emilia coccinea, Hibiscus rosasinensis, and Asystasia gangetica to act as superoxide
radicals (SOR), hydrogen peroxide (HP), nitric oxide radical (NOR), hydroxyl radical (HR), and 2,2-
diphenyl-1-picrylhydrazyl (DPPH) radical antagonists using in vitro models. The herbal extracts were
single herbal formulations (SHfs), double herbal formulations (DHfs), triple herbal formulations (THfs),
and a quadruple herbal formulation (QHf). The phytochemical composition and radical scavenging capacity
index (SCI) of the herbal formulations were measured using standard methods. The flavonoids
were the most abundant phytochemicals present in the herbal extracts. The SCI50 defined the concentration
(mg/mL) of herbal formulation required to scavenge 50% of the investigated radicals. The SHfs,
DHfs, THfs, and QHf SCI50 against the radicals followed the order HR > SOR > DPPH radical > HP > NOR.
Although the various herbal formulations exhibited ambivalent antioxidant activities in terms of their
radical scavenging capabilities, a broad survey of the results of the present study showed that combinatorial
herbal formulations (DHfs, THfs, and QHf) appeared to exhibit lower radical scavenging capacities
than those of the SHfs in vitro.
Effects of foliar application with salicylic acid on the biochemical paramete...INNS PUBNET
Low temperature is an important environmental factor that limits the survival, productivity and geographical distribution of plants. Oil seeds are the second global food resources among which Brassica napus L. is the third annual oil seed in the world. In cold stress, some biochemical and physiological reactions occur in response to reactive oxygen species (ROS). Hence, the effect of foliar application of salicylic acid (SA) on total chlorophyll content, malondialdehyde, and antioxidant enzymes activity and solute protein and proline contents were assessed in two canola varieties (Brassica napus L., cv RGS and LICORD) leaves exposed to cold stress during 0, 24, and 48 hours after salicylic acid treatment. They were first grown in a controlled growth room at 22/20 °C (day/night) for one month followed by SA spraying application (100, 200 and 400µM) and then plots were transferred to a cold environment (-2 °C) for 3 days. The results showed that the total chlorophyll content was decreased in RGS cultivars related to high salicylic acid concentration during the experiment. The results of antioxidant status showed that superoxide dismutase (SOD), peroxidase (POX), and also lipid peroxidation were increased significantly after 48 hours compared first day. Catalase (CAT) activity was decreased 24 hours after salicylic acid treatment. Results showed an increase in protein content in both cultivars treated with SA, by contrast proline was greatly affected by salicylic acid treatment and its content was the highest 24 hours after treatment. According to the results of the present study indicated that application of salicylic acid has useful effects on the biochemical traits. Thereupon it may be effective for the improvement of plant growth in cold regions.
DPPH Scavenging Assay of Eighty Four Bangladeshi Medicinal PlantsIOSR Journals
This study was designed to screen out free radical scavenging potentiality of 84 medicinal plants. Stock solution of different plant extracts and standard were diluted to achieve suitable concentrations. A control was also prepared without plant extract solution. Then 0.004% DPPH solution was added. The mixtures were incubated in the room temperature for 30 minutes. Then the absorbance was measured at 517 nm against solvent in UV-spectrophotometer and then IC50 was calculated. In this experiment two standard were used-ascorbic acid and BHT. Both showed a significant IC50 value of 15.5μg/mL, and 46.54μg/mL respectively. Among 84 medicinal plants Syzygim cumini, Casuarina littorea, Borassus flabellifer, Enhydra fluctuans, and Minusops elengi exhibited highest radical scavenging potential with an IC50 value of 12.816μg/mL, 14.467μg/mL, 15.755μg/mL, 15.653μg/mL, and 20.380μg/mL respectively. All these value are very close to the IC50 value of ascorbic acid and better than IC50 value of BHT (Butylated Hydroxy Toluene). Syzygim cumini is the most powerful scavenger among all tested medicinal plants and also most strong scavenger than ascorbic acid and BHT. Scavenging activity was found to increase in dose dependent manner. Another 30 medicinal plants exhibited good scavenging property and 14 medicinal plants showed moderate scavenging activity. The rest presented lower scavenging activity. This present study indicates that plants having good scavenging property may have various health beneficial effects and these plants can be considered as valuable source of bioactive components with high antioxidant properties.
Hepatoprotective and antioxidant effects of Azolla microphylla based gold nan...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Our present study sought to evaluate hepatoprotective and antioxidant effects of methanol extract of Azolla microphylla phytochemically synthesized gold nanoparticles (GNaP) in acetaminophen (APAP) - induced hepatotoxicity of fresh water common carp fish.
Materials and Methods:
GNaP were prepared by green synthesis method using methanol extract of Azolla microphylla. Twenty four fishes weighing 146 ± 2.5 g were used in this experiment and these were divided into four experimental groups, each comprising 6 fishes. Group 1 served as control. Group 2 fishes were exposed to APAP (500 mg/kg) for 24 h. Groups 3 and 4 fishes were exposed to APAP (500 mg/kg) + GNaP (2.5 mg/kg) and GNaP (2.5 mg/kg) for 24 h, respectively. The hepatoprotective and antioxidant potentials were assessed by measuring liver damage, biochemical parameters, ions status, and histological alterations.
Results:
APAP exposed fish showed significant elevated levels of metabolic enzymes (LDH, G6PDH and MDH), hepatotoxic markers (GPT, GOT and ALP), reduced hepatic glycogen, lipids, protein, albumin, globulin, increased levels of bilirubin, creatinine, and oxidative stress markers (TBRAS, LHP and protein carbonyl), altered the tissue enzymes (SOD, CAT, GSH-Px and GST) non-enzyme (GSH), cellular sulfhydryl (T-SH, P-SH and NP-SH) levels, reduced hepatic ions (Ca2+, Na+ and K+), and abnormal liver histology. It was observe that GNaP has reversal effects on the levels of above mentioned parameters in APAP hepatotoxicity.
Conclusion:
Azolla microphylla phytochemically synthesized GNaP protects liver against oxidative damage and tissue damaging enzyme activities and could be used as an effective protector against acetaminophen-induced hepatic damage in fresh water common carp fish.
In Vitro Cell Tests for Functional FoodInstitut Kurz
The relationship between the food we eat and our health is
clear. In the constant search for healthier foods rich in
bioactive compounds that promote health and healthy
ageing, a wide variety of functional foods have appeared on
the market.
To know the real function of these functional foods
in our body, it is necessary to carry out different types of in
vitro cell tests.
Institut Kurz specializes in conducting in vitro cell tests for functional foods.
Contact us for more information:
info@institut-kurz.com
https://www.institut-kurz.com/
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Engineering escherichia coli to convert acetic acid to free fatty acidszhenhua82
Fatty acids (FAs) are promising precursors of advanced biofuels. This study investigated conversion of acetic acid (HAc) to FAs by an engineered Escherichia coli strain. We combined established genetic engineering strategies including overexpression of acs and tesA genes, and knockout of fadE in E. coli BL21, resulting in the production of similar to 1 g/L FAs from acetic acid. The microbial conversion of HAc to FAs was achieved with similar to 20% of the theoretical yield. We cultured the engineered strain with HAc-rich liquid wastes, which yielded similar to 0.43 g/L FAs using waste streams from dilute acid hydrolysis of lignocellulosic biomass and similar to 0.17 g/L FAs using effluent from anaerobic-digested sewage sludge. C-13-isotopic experiments showed that the metabolism in our engineered strain had high carbon fluxes toward FAs synthesis and TCA cycle in a complex HAc medium. This proof-of-concept work demonstrates the possibility for coupling the waste treatment with the biosynthesis of advanced biofuel via genetically engineered microbial species.
Reactive oxygen and nitrogen species (RONS) are involved in deleterious/beneficial biological processes.
The present study sought to investigate the capacity of single and combinatorial herbal formulations of
Acanthus montanus, Emilia coccinea, Hibiscus rosasinensis, and Asystasia gangetica to act as superoxide
radicals (SOR), hydrogen peroxide (HP), nitric oxide radical (NOR), hydroxyl radical (HR), and 2,2-
diphenyl-1-picrylhydrazyl (DPPH) radical antagonists using in vitro models. The herbal extracts were
single herbal formulations (SHfs), double herbal formulations (DHfs), triple herbal formulations (THfs),
and a quadruple herbal formulation (QHf). The phytochemical composition and radical scavenging capacity
index (SCI) of the herbal formulations were measured using standard methods. The flavonoids
were the most abundant phytochemicals present in the herbal extracts. The SCI50 defined the concentration
(mg/mL) of herbal formulation required to scavenge 50% of the investigated radicals. The SHfs,
DHfs, THfs, and QHf SCI50 against the radicals followed the order HR > SOR > DPPH radical > HP > NOR.
Although the various herbal formulations exhibited ambivalent antioxidant activities in terms of their
radical scavenging capabilities, a broad survey of the results of the present study showed that combinatorial
herbal formulations (DHfs, THfs, and QHf) appeared to exhibit lower radical scavenging capacities
than those of the SHfs in vitro.
Effects of foliar application with salicylic acid on the biochemical paramete...INNS PUBNET
Low temperature is an important environmental factor that limits the survival, productivity and geographical distribution of plants. Oil seeds are the second global food resources among which Brassica napus L. is the third annual oil seed in the world. In cold stress, some biochemical and physiological reactions occur in response to reactive oxygen species (ROS). Hence, the effect of foliar application of salicylic acid (SA) on total chlorophyll content, malondialdehyde, and antioxidant enzymes activity and solute protein and proline contents were assessed in two canola varieties (Brassica napus L., cv RGS and LICORD) leaves exposed to cold stress during 0, 24, and 48 hours after salicylic acid treatment. They were first grown in a controlled growth room at 22/20 °C (day/night) for one month followed by SA spraying application (100, 200 and 400µM) and then plots were transferred to a cold environment (-2 °C) for 3 days. The results showed that the total chlorophyll content was decreased in RGS cultivars related to high salicylic acid concentration during the experiment. The results of antioxidant status showed that superoxide dismutase (SOD), peroxidase (POX), and also lipid peroxidation were increased significantly after 48 hours compared first day. Catalase (CAT) activity was decreased 24 hours after salicylic acid treatment. Results showed an increase in protein content in both cultivars treated with SA, by contrast proline was greatly affected by salicylic acid treatment and its content was the highest 24 hours after treatment. According to the results of the present study indicated that application of salicylic acid has useful effects on the biochemical traits. Thereupon it may be effective for the improvement of plant growth in cold regions.
DPPH Scavenging Assay of Eighty Four Bangladeshi Medicinal PlantsIOSR Journals
This study was designed to screen out free radical scavenging potentiality of 84 medicinal plants. Stock solution of different plant extracts and standard were diluted to achieve suitable concentrations. A control was also prepared without plant extract solution. Then 0.004% DPPH solution was added. The mixtures were incubated in the room temperature for 30 minutes. Then the absorbance was measured at 517 nm against solvent in UV-spectrophotometer and then IC50 was calculated. In this experiment two standard were used-ascorbic acid and BHT. Both showed a significant IC50 value of 15.5μg/mL, and 46.54μg/mL respectively. Among 84 medicinal plants Syzygim cumini, Casuarina littorea, Borassus flabellifer, Enhydra fluctuans, and Minusops elengi exhibited highest radical scavenging potential with an IC50 value of 12.816μg/mL, 14.467μg/mL, 15.755μg/mL, 15.653μg/mL, and 20.380μg/mL respectively. All these value are very close to the IC50 value of ascorbic acid and better than IC50 value of BHT (Butylated Hydroxy Toluene). Syzygim cumini is the most powerful scavenger among all tested medicinal plants and also most strong scavenger than ascorbic acid and BHT. Scavenging activity was found to increase in dose dependent manner. Another 30 medicinal plants exhibited good scavenging property and 14 medicinal plants showed moderate scavenging activity. The rest presented lower scavenging activity. This present study indicates that plants having good scavenging property may have various health beneficial effects and these plants can be considered as valuable source of bioactive components with high antioxidant properties.
Hepatoprotective and antioxidant effects of Azolla microphylla based gold nan...Nanomedicine Journal (NMJ)
Abstract
Objective(s):
Our present study sought to evaluate hepatoprotective and antioxidant effects of methanol extract of Azolla microphylla phytochemically synthesized gold nanoparticles (GNaP) in acetaminophen (APAP) - induced hepatotoxicity of fresh water common carp fish.
Materials and Methods:
GNaP were prepared by green synthesis method using methanol extract of Azolla microphylla. Twenty four fishes weighing 146 ± 2.5 g were used in this experiment and these were divided into four experimental groups, each comprising 6 fishes. Group 1 served as control. Group 2 fishes were exposed to APAP (500 mg/kg) for 24 h. Groups 3 and 4 fishes were exposed to APAP (500 mg/kg) + GNaP (2.5 mg/kg) and GNaP (2.5 mg/kg) for 24 h, respectively. The hepatoprotective and antioxidant potentials were assessed by measuring liver damage, biochemical parameters, ions status, and histological alterations.
Results:
APAP exposed fish showed significant elevated levels of metabolic enzymes (LDH, G6PDH and MDH), hepatotoxic markers (GPT, GOT and ALP), reduced hepatic glycogen, lipids, protein, albumin, globulin, increased levels of bilirubin, creatinine, and oxidative stress markers (TBRAS, LHP and protein carbonyl), altered the tissue enzymes (SOD, CAT, GSH-Px and GST) non-enzyme (GSH), cellular sulfhydryl (T-SH, P-SH and NP-SH) levels, reduced hepatic ions (Ca2+, Na+ and K+), and abnormal liver histology. It was observe that GNaP has reversal effects on the levels of above mentioned parameters in APAP hepatotoxicity.
Conclusion:
Azolla microphylla phytochemically synthesized GNaP protects liver against oxidative damage and tissue damaging enzyme activities and could be used as an effective protector against acetaminophen-induced hepatic damage in fresh water common carp fish.
In Vitro Cell Tests for Functional FoodInstitut Kurz
The relationship between the food we eat and our health is
clear. In the constant search for healthier foods rich in
bioactive compounds that promote health and healthy
ageing, a wide variety of functional foods have appeared on
the market.
To know the real function of these functional foods
in our body, it is necessary to carry out different types of in
vitro cell tests.
Institut Kurz specializes in conducting in vitro cell tests for functional foods.
Contact us for more information:
info@institut-kurz.com
https://www.institut-kurz.com/
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Engineering escherichia coli to convert acetic acid to free fatty acidszhenhua82
Fatty acids (FAs) are promising precursors of advanced biofuels. This study investigated conversion of acetic acid (HAc) to FAs by an engineered Escherichia coli strain. We combined established genetic engineering strategies including overexpression of acs and tesA genes, and knockout of fadE in E. coli BL21, resulting in the production of similar to 1 g/L FAs from acetic acid. The microbial conversion of HAc to FAs was achieved with similar to 20% of the theoretical yield. We cultured the engineered strain with HAc-rich liquid wastes, which yielded similar to 0.43 g/L FAs using waste streams from dilute acid hydrolysis of lignocellulosic biomass and similar to 0.17 g/L FAs using effluent from anaerobic-digested sewage sludge. C-13-isotopic experiments showed that the metabolism in our engineered strain had high carbon fluxes toward FAs synthesis and TCA cycle in a complex HAc medium. This proof-of-concept work demonstrates the possibility for coupling the waste treatment with the biosynthesis of advanced biofuel via genetically engineered microbial species.
Inria - leaflet of research centre Saclay - Île-de-FranceInria
The Inria Saclay – Île-de-France research centre is a major part of the highly diversified ecosystem at Saclay which features leading partners in computational sciences. Research bodies, leading schools, competitiveness clusters and industry comprise the three complementary facets of research, training and innovation. A coordination program launched in 2000 took concrete form with the establishment of the Digiteo advanced research network. Most of our research teams are pooled with several institutions as proof of the strong links we enjoy with our partners. Centre researchers contribute to education for the students in Saclay. Training through research extends the approach with many doctoral students welcomed in research teams.
Construction of a 200' long x 16' high x 3' thick x 3500 psi, solid concrete retaining wall, backfilled with 3000m3 of road grade stone, compacted every 12"
CONNECT, the magazine for SMEs seeking innovation through the digital sciences, and get access to ICST news, updates on the state of research and applied technology for industry, portraits of partners who have participated in INRIA research teams working with innovative SMEs, special reports and other features. In the current issue: Is cloud computing where it's at for SMEs? Digital sciences for domestic health.
Role of Oxidative Stress on male infertilityFavourji
Oxidative stress is associated with several body cell damage which can result into cell death and eventually mortality. Its a key factor in several infertility cases and with the knowledge of its impact a lot can be known to avert its further implications.
LABORATORY ANALYSIS OF ANTIOXIDANTS IN FOODInstitut Kurz
To characterize the antioxidants in food, it is necessary to know:
- How much antioxidant is contained in the food.
- Are the antioxidants absorbed from the food to the blood?
- If the antioxidants in the blood make their way to the intended body cells.
- Do the antioxidants help the cells to defend themselves against oxygen radicals? May it be given by increasing the production of the cell's own defense enzymes or by direct chemical neutralization of the radicals?
iNSTITUT KURZ performs all laboratory analysis of antioxidants in food!
VISIT OUR WEBSITE: www.institut-kurz.com/
Toxicology testing, also known as safety assessment, or toxicity testing, is conducted to determine the degree to which a substance can damage a living or non-living organism. It is often conducted by researchers using standard test procedures to comply with governing regulations, for example for medicines and pesticides. Much toxicology is considered to be part of the field of preclinical development. Stages of in vitro and in vivo research are conducted to determine safe doses of exposure in humans before a first-in-man study. Toxicology testing may be conducted by the pharmaceutical industry, biotechnology companies or contract research organizations.
Drug development is a high-risk enterprise. The typical new drug takes 10-12 years to get to market and costs up to $500 million. Pharmaceutical companies face continually increasing challenges in drug development— shorter product life cycles, global competition, as well as daunting technical and regulatory hurdles. Meanwhile, as a result of the Human Genome Project and high throughput drug development methods, there are many more drug candidates to test. Thus, there is growing pressure on pharmaceutical and biopharmaceutical companies.
Antioxidant Activity and Cytotoxicity against Cancer Cell Lines of the Extrac...Juan Hernandez
Xylaria species associated with termite nests or soil have been considered rare species in nature and the few which have been reported upon have been found to act as a rich source of bioactive metabolites. This study evaluated 10 ethyl acetate extracts of five new Xylaria species associated with termite nests or soil for their antioxidant activity, and cytotoxicity against different cancer and normal cell lines. DPPH and ABTS radical scavenging activities of the extracts demonstrated strong capacity with low IC50 values. The highest observed activities belonged to X. vinacea SWUF18-2.3 having IC50 values of 0.194 ± 0.031 mg/mL for DPPH assay and 0.020 ± 0.004 mg/mL for ABTS assay. Total phenolic content ranged from 0.826 ± 0.123 to 3.629 ± 0.381 g GAE/g crude extract which correlated with antioxidant activities. The high total phenolic content could contribute to the high antioxidant activities. Cytotoxicity was recorded against A549, HepG2, HeLa and PNT2 and resulted in broad spectrum to specific activity depending on the cell lines. The highest activities were observed with X. subintraflava SWUF16-11.1 which resulted in 11.15 ± 0.32 to 13.17 ± 2.37% cell viability at a concentration of 100 µg/mL. Moreover, LC-MS fingerprints indicated over 61 peaks from all isolates. There were 18 identified and 43 unidentified compounds compared to mass databases. The identified compounds were from various groups of diterpenoids, diterpenes, cytochalasin, flavones, flavonoids, polyphenols, steroids and derivatives, triterpenoids and tropones. These results indicate that Xylaria spp. has abundant secondary metabolites that could be further explored for their therapeutic properties
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
Nutrients present in Nutrease powder play an important role in maintaining the normal functions of the human body. The major nutrients present in Nutrease powder include Natural carbohydrates, proteins, lipids, vitamins, and minerals. Besides these, there are some bioactive food components known as “phytonutrients” that play an important role in human health. They have tremendous impact on the health care system and may provide medical health benefits including the prevention and/or treatment of disease and various physiological disorders such as Andropause or Male menopause. Phytonutrients play a positive role by maintaining and modulating immune function to prevent specific diseases. Being natural products, they hold a great promise in clinical therapy. Phytonutrients in Nutrease powder are the plant nutrients with specific biological activities that support human health. Some of the important bioactive phytonutrients in Nutrease powder include polyphenols, terpenoids, resveratrol, flavonoids, isoflavonoids, carotenoids, limonoids, glucosinolates, phytoestrogens, phytosterols, anthocyanins, and probiotics. They play specific pharmacological effects in human health such as anti-microbial, anti-oxidants, anti-inflammatory, anti-allergic, anti-spasmodic, anti-cancer, anti-aging, hepatoprotective, hypolipidemic, neuroprotective, hypotensive, diabetes, osteoporosis, CNS stimulant, analgesic, protection from UVB-induced carcinogenesis, immuno-modulator, and carminative. This article reviews the current available scientific literature regarding the effect of Nutrease powder as an effective supplementation in Male & Female Fertility.
Antioxidant potentials of tannic acid on lipid peroxidation induced by severa...Premier Publishers
Various prospective studies have indicated the antioxidant potency of tannic acid in several models. However, there is no clear-cut evidence revealing that the reported antioxidant properties of tannic acid remains potent regardless of the lipid sources and pro-oxidants employed for the oxidative assault. Hence, this study sought to investigate the antioxidant properties of tannic acid against cerebral and hepatic lipid peroxidation induced by several pro-oxidants (Iron (II) sulfate, Sodium nitroprusside, cyclophosphamide and acetaminophen) in vitro. Rats were decapitated under mild ether anesthesia and the tissues were rapidly dissected, placed on ice, weighed and immediately homogenized in cold 50 mM Tris-HCl, pH 7.4 (1/10, w/v). The homogenates were centrifuged for 10 min at 4000 g to yield a pellet that was discarded and a low-speed supernatant (S1). Our results indicated that Fe (II) showed the highest pro-oxidative effects in both tissues lipids. Furthermore, tannic acid demonstrated potent inhibitory effects against lipid peroxidation in both tissues lipids regardless of the pro-oxidant employed. To this end, there is a dire need to exploit the protective benefits of tannic acid as a potential exogenous antioxidant against lipid peroxidation with a view to providing solution to the global oxidative stress menace.
The Indian Dental Academy is the Leader in continuing dental education , training dentists in all aspects of dentistry and
offering a wide range of dental certified courses in different formats.for more details please visit
www.indiandentalacademy.com
Transcript: Selling digital books in 2024: Insights from industry leaders - T...BookNet Canada
The publishing industry has been selling digital audiobooks and ebooks for over a decade and has found its groove. What’s changed? What has stayed the same? Where do we go from here? Join a group of leading sales peers from across the industry for a conversation about the lessons learned since the popularization of digital books, best practices, digital book supply chain management, and more.
Link to video recording: https://bnctechforum.ca/sessions/selling-digital-books-in-2024-insights-from-industry-leaders/
Presented by BookNet Canada on May 28, 2024, with support from the Department of Canadian Heritage.
Slack (or Teams) Automation for Bonterra Impact Management (fka Social Soluti...Jeffrey Haguewood
Sidekick Solutions uses Bonterra Impact Management (fka Social Solutions Apricot) and automation solutions to integrate data for business workflows.
We believe integration and automation are essential to user experience and the promise of efficient work through technology. Automation is the critical ingredient to realizing that full vision. We develop integration products and services for Bonterra Case Management software to support the deployment of automations for a variety of use cases.
This video focuses on the notifications, alerts, and approval requests using Slack for Bonterra Impact Management. The solutions covered in this webinar can also be deployed for Microsoft Teams.
Interested in deploying notification automations for Bonterra Impact Management? Contact us at sales@sidekicksolutionsllc.com to discuss next steps.
Connector Corner: Automate dynamic content and events by pushing a buttonDianaGray10
Here is something new! In our next Connector Corner webinar, we will demonstrate how you can use a single workflow to:
Create a campaign using Mailchimp with merge tags/fields
Send an interactive Slack channel message (using buttons)
Have the message received by managers and peers along with a test email for review
But there’s more:
In a second workflow supporting the same use case, you’ll see:
Your campaign sent to target colleagues for approval
If the “Approve” button is clicked, a Jira/Zendesk ticket is created for the marketing design team
But—if the “Reject” button is pushed, colleagues will be alerted via Slack message
Join us to learn more about this new, human-in-the-loop capability, brought to you by Integration Service connectors.
And...
Speakers:
Akshay Agnihotri, Product Manager
Charlie Greenberg, Host
Let's dive deeper into the world of ODC! Ricardo Alves (OutSystems) will join us to tell all about the new Data Fabric. After that, Sezen de Bruijn (OutSystems) will get into the details on how to best design a sturdy architecture within ODC.
LF Energy Webinar: Electrical Grid Modelling and Simulation Through PowSyBl -...DanBrown980551
Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
Then welcome to this PowSyBl workshop, hosted by Rte, the French Transmission System Operator (TSO)!
During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
- A fully editable and extendable library for grid component modelling;
- Visualization tools to display your network;
- Grid simulation tools, such as power flows, security analyses (with or without remedial actions) and sensitivity analyses;
The framework is mostly written in Java, with a Python binding so that Python developers can access PowSyBl functionalities as well.
What you will learn during the webinar:
- For beginners: discover PowSyBl's functionalities through a quick general presentation and the notebook, without needing any expert coding skills;
- For advanced developers: master the skills to efficiently apply PowSyBl functionalities to your real-world scenarios.
Dev Dives: Train smarter, not harder – active learning and UiPath LLMs for do...UiPathCommunity
💥 Speed, accuracy, and scaling – discover the superpowers of GenAI in action with UiPath Document Understanding and Communications Mining™:
See how to accelerate model training and optimize model performance with active learning
Learn about the latest enhancements to out-of-the-box document processing – with little to no training required
Get an exclusive demo of the new family of UiPath LLMs – GenAI models specialized for processing different types of documents and messages
This is a hands-on session specifically designed for automation developers and AI enthusiasts seeking to enhance their knowledge in leveraging the latest intelligent document processing capabilities offered by UiPath.
Speakers:
👨🏫 Andras Palfi, Senior Product Manager, UiPath
👩🏫 Lenka Dulovicova, Product Program Manager, UiPath
The Art of the Pitch: WordPress Relationships and SalesLaura Byrne
Clients don’t know what they don’t know. What web solutions are right for them? How does WordPress come into the picture? How do you make sure you understand scope and timeline? What do you do if sometime changes?
All these questions and more will be explored as we talk about matching clients’ needs with what your agency offers without pulling teeth or pulling your hair out. Practical tips, and strategies for successful relationship building that leads to closing the deal.
Accelerate your Kubernetes clusters with Varnish CachingThijs Feryn
A presentation about the usage and availability of Varnish on Kubernetes. This talk explores the capabilities of Varnish caching and shows how to use the Varnish Helm chart to deploy it to Kubernetes.
This presentation was delivered at K8SUG Singapore. See https://feryn.eu/presentations/accelerate-your-kubernetes-clusters-with-varnish-caching-k8sug-singapore-28-2024 for more details.
Essentials of Automations: Optimizing FME Workflows with ParametersSafe Software
Are you looking to streamline your workflows and boost your projects’ efficiency? Do you find yourself searching for ways to add flexibility and control over your FME workflows? If so, you’re in the right place.
Join us for an insightful dive into the world of FME parameters, a critical element in optimizing workflow efficiency. This webinar marks the beginning of our three-part “Essentials of Automation” series. This first webinar is designed to equip you with the knowledge and skills to utilize parameters effectively: enhancing the flexibility, maintainability, and user control of your FME projects.
Here’s what you’ll gain:
- Essentials of FME Parameters: Understand the pivotal role of parameters, including Reader/Writer, Transformer, User, and FME Flow categories. Discover how they are the key to unlocking automation and optimization within your workflows.
- Practical Applications in FME Form: Delve into key user parameter types including choice, connections, and file URLs. Allow users to control how a workflow runs, making your workflows more reusable. Learn to import values and deliver the best user experience for your workflows while enhancing accuracy.
- Optimization Strategies in FME Flow: Explore the creation and strategic deployment of parameters in FME Flow, including the use of deployment and geometry parameters, to maximize workflow efficiency.
- Pro Tips for Success: Gain insights on parameterizing connections and leveraging new features like Conditional Visibility for clarity and simplicity.
We’ll wrap up with a glimpse into future webinars, followed by a Q&A session to address your specific questions surrounding this topic.
Don’t miss this opportunity to elevate your FME expertise and drive your projects to new heights of efficiency.
PHP Frameworks: I want to break free (IPC Berlin 2024)Ralf Eggert
In this presentation, we examine the challenges and limitations of relying too heavily on PHP frameworks in web development. We discuss the history of PHP and its frameworks to understand how this dependence has evolved. The focus will be on providing concrete tips and strategies to reduce reliance on these frameworks, based on real-world examples and practical considerations. The goal is to equip developers with the skills and knowledge to create more flexible and future-proof web applications. We'll explore the importance of maintaining autonomy in a rapidly changing tech landscape and how to make informed decisions in PHP development.
This talk is aimed at encouraging a more independent approach to using PHP frameworks, moving towards a more flexible and future-proof approach to PHP development.
GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
1. J. Agric. Food Chem. 2008, 56, 8319–8325 8319
Comparison of Chemical and Cell-Based Antioxidant
Methods for Evaluation of Foods and Natural
Products: Generating Multifaceted Data by Parallel
Testing Using Erythrocytes and Polymorphonuclear
Cells
DANA HONZEL,† STEVE G. CARTER,† KIMBERLEE A. REDMAN,†
ALEXANDER G. SCHAUSS,‡ JOHN R. ENDRES,‡ AND GITTE S. JENSEN*,§
NIS Labs, 1437 Esplanade, Klamath Falls, Oregon 97601; Natural & Medicinal Products Research,
AIBMR Life Sciences Inc., Puyallup, Washington 98373; and Holger NIS Inc.,
601 13th Avenue N.E., Calgary, Alberta, Canada T2E 1C7
The objective of this study was to compare three tests frequently used for evaluation of antioxidant
potential in natural products: (1) oxygen radical absorbance assay (ORAC), (2) cell-based antioxidant
protection in an erythrocyte model (CAP-e), and (3) reactive oxygen species formation in polymor-
phonuclear cells (ROS PMN). The methods were applied to four natural products, all containing
antioxidants capable of entering and protecting cells in the CAP-e assay. The magnitude of this effect
was not directly correlated to the ORAC value of each product. Furthermore, the products showed
different effects in the ROS PMN assay. Acai provided strong inhibition of ROS formation, indicating
¸
anti-inflammatory properties. In contrast, Immunel and EpiCor mildly enhanced ROS formation,
suggesting activation of the innate immune response. HA Joint Formula showed a complex, nonlinear
dose-response in the ROS PMN assay. This illustrates that complex natural products may have similar
antioxidant properties but different effects on human cells. Cell-based antioxidant protection is
addressed best in the CAP-e assay, since some natural products contain compounds that may provoke
cellular signaling in other cell types. The PMN cell type is a useful model for assessment of overall
anti-inflammatory versus immune supportive properties of a product. The sequential use of the three
methods serves to bridge analytical and biological testing methods.
KEYWORDS: Antioxidant; method; erythrocyte; polymorphonuclear; cell-based antioxidant protection
assay (CAP-e); oxygen radical absorbance capacity (ORAC) assay
INTRODUCTION been associated with obesity (2), immune dysfunction (3),
cardiovascular disease (4), declining cognitive function (5), and
Free radicals arise during normal metabolism, are produced
cancer (6, 7).
during immune activity, and are introduced by many environ-
mental factors such as pollution, smoke, and sunlight. Antioxi- As continuing research examines the effect of antioxidants
dant mechanisms in blood, cells, and tissue fluids play an on health, the testing for antioxidant protection has become a
important role in neutralizing the normal level of oxidative powerful focus in the dietary and natural products industry.
damage caused by free radicals. During chronic inflammatory Researchers associated with the natural product industry have
conditions, combined with the absence of sufficient dietary pushed for a standardized method for measuring antioxidant
antioxidants, the oxidative damage is accelerated, causing further capacity in natural products (8, 9). A large number of methods
dysregulation involving inflammatory reactions that contribute have been developed to evaluate the antioxidant capacity in
to many degenerative diseases and aging (1). A growing body foods, including nutritional supplements, vitamins, minerals, and
of research is focused on nutritional and pharmaceutical extracts of various natural products of plant, fungal, animal,
prevention of chronic inflammatory conditions, as these have and bacterial origins. The natural products industry has taken
many steps in creating standardized tests to measure antioxidant
levels in (1) nutritional and natural products and (2) blood
* To whom correspondence should be addressed. Phone: (403) 277-
4134. Fax: (403) 441-5236. E-mail: gitte@holgernis.com.
samples before and after consuming such products. One of the
†
NIS Labs. most popular and best standardized chemical antioxidant
‡
AIBMR Life Sciences Inc. methods is the oxygen radical absorbance capacity (ORAC)
§
Holger NIS Inc. test (10–12). This test is widely used for evaluation and
10.1021/jf800401d CCC: $40.75 2008 American Chemical Society
Published on Web 08/22/2008
2. 8320 J. Agric. Food Chem., Vol. 56, No. 18, 2008 Honzel et al.
Table 1. Comparison of Chemical and Cell-Based Assays in Vitro
chemical assays cell-based assays
assay principles based on known chemical reactions based on interaction between added
between limited number of reagents compounds and complex enzymatic
reactions in biological system
extraction of active ingredients some flexibility assay must take place in physiological
saline solution; limited use of solvents
use of alcohol-based solvents yes, if no interference with yes, if properly diluted, tested for tolerance,
chemical reactions in assay and altered cellular behavior (depends on assay)
dimethyl sulfoxide (DMSO) yes, with appropriate controls, if no no, alters bioavailability (14–16),
as solvent interference with chemical reactions in assay; thereby defeating the purpose of testing
DMSO is a free radical scavenger (17) bioavailability in vitro; DMSO is anti-inflammatory (18) and can
exaggerate mitochondrial ROS formation (19)
data analysis and interpretation quantitative qualitative
expectation of linear dose- responses yes no
applicability of area-under-curve yes (11) no
Table 2. Enzymes Involved in Redox Reactions in Red Blood Cells (RBC) and Polymorphonuclear Cells (PMN)
enzyme role for normal cell physiology RBC PMN
glutathione peroxidase reduces lipid hydroperoxides to their corresponding alcohols; reduces free X X
hydrogen peroxide to water; protects integrity of cellular membranes; function
depends on selenium; protects nucleated cells from oxidative stress-induced
programmed cell death (apoptosis)
glutathione reductase maintains glutathione in its reduced state X X
catalase breakdown of hydrogen peroxide to oxygen and water X X
superoxide dismutase catalyzes dismutation of superoxide to oxygen and hydrogen peroxide X X
myeloperoxidase produces hypochlorous acid (HOCl) from hydrogen peroxide (H2O2) and X
chloride anion (Cl-) during the neutrophil’s respiratory burst; oxidizes tyrosine
to tyrosyl radical using hydrogen peroxide
cyclooxygenase COX-1 contain both cyclooxygenase and peroxidase properties; catalyzes the first step constitutively expressed
in the biosynthesis of prostaglandins, thromboxanes, and prostacyclins
cyclooxygenase COX-2 contain both cyclooxygenase and peroxidase properties; catalyzes the first step induced upon inflammation
in the biosynthesis of prostaglandins, thromboxanes, and prostacyclins
hemoglobin/deoxyhemoglobin scavenging and transport of O2, CO2, NO- X
comparison of the antioxidant capacity in natural products and We have developed the CAP-e assay (21) as a cell-based
extracts and has been successfully applied to bioavailability antioxidant protection assay using erythrocytes to address the
studies, showing increased antioxidant capacity in serum in test question of whether antioxidants in complex natural products
subjects after consumption of antioxidant-rich foods (13). enter the cytosol and contribute to the reduction of oxidative
Going beyond the ORAC test for antioxidant testing currently damage within the cell (Figure 1). The assay measures the
requires either more complex cellular testing systems or full effects in the cytosol only, as the reporter dye we use in the
clinical trials focused on serum antioxidant capacity, lipid test is only functional after penetrating into the intracellular
peroxidation, and associated physiological measurements. The space (i.e., the cytosol) where it undergoes chemical modifica-
expense of full clinical trials makes selected panels of cell-based tion, resulting in its retention within the cell. The assay allows
assays attractive as intermediate testing methods. The transition for semiquantification specifically of those antioxidants that are
from chemical to cell-based assays involves different solubility capable of penetrating into live cells. We used this test as a
issues and restricted use of solvents, particularly DMSO, which baseline for further cell-based testing using primary pro-
alters bioavailability (14–16) and possesses antioxidant and anti- inflammatory PMN cells, to allow for a more definitive
inflammatory properties (17–19) (Table 1). assessment of the complex properties of natural products in vitro.
The use of erythrocytes, or red blood cells (RBC), in testing Examples of other existing cell-based assays for measuring
how well a substance protects against oxidative stress is effects on inflammatory cell types includes testing on freshly
biologically relevant, as RBC play critical roles in antioxidant isolated human PMN cells (22, 23). This cell type is an important
protection in the blood (20). The ability of RBC to scavenge part of our innate immune defense and is capable of rapid
reactive oxygen and nitrogen species represents a direct production of ROS in response to both oxidative damage and pro-
antioxidant and anti-inflammatory protection to the body. The inflammatory stimuli. The PMN cell can respond to compounds
cells are living, energy-producing cells and represent the most in natural products extracts in three distinct ways; the data obtained
abundant cell type in the blood circulation where they from a PMN-based assay represent a total summary of these
outnumber white blood cells by at least 100-fold. Using RBC mechanisms often operating in parallel (Figure 1). Thus, data
as a cell model reduces the confounding contribution of obtained from a PMN-based assay may be interpreted better in
cellular signaling. The assay is not affected by mitochondrial the light of a preceding RBC-based assay, such as the CAP-e.
production of reactive oxygen species (ROS), as RBC do
not have mitochondria but instead generate cellular energy MATERIALS AND METHODS
by other metabolic pathways. Many of the redox enzymes Reagents. The following buffers and reagents were obtained from
present in RBC are the same as the enzymes present in Sigma-Aldrich (St. Louis, MO): phosphate-buffered saline (PBS),
polymorphonuclear (PMN) cells (Table 2). RPMI-1640 culture medium, hydrogen peroxide 30% solution (H2O2),
3. Antioxidant Methods for Foods and Natural Products J. Agric. Food Chem., Vol. 56, No. 18, 2008 8321
Evaluation of Antioxidant Protection in a Red Blood Cell-Based
Assay. The red blood cell is a convenient test model for examination
of antioxidants that are able to enter living cells. Since the red blood
cell is not able to signal in response to pro-inflammatory stimuli and is
not able to produce reactive oxygen species, it provides a cleaner signal
for antioxidant capacity of a test product than the PMN cell. Whole
blood was layered onto Histopaque 1119 and centrifuged 25 min at
2400 rpm. All plasma, leukocytes, and Histopaque were removed. From
the remaining packed red blood cells, 0.1 mL was transferred into 10
mL PBS without calcium or magnesium. Parallel samples of this red
blood cell suspension were incubated at 37 °C, 5% CO2 for 90 min,
either untreated or with test products over a range of 5-fold serial
dilutions from 10 to 0.016 mg/mL. A stock solution of DCF-DA, which
becomes brightly green fluorescent upon exposure to free radicals, was
prepared by adding 0.18 mL of DMSO to a 0.05 mg aliquot of DCF-
DA and vortexing 3 times for 15 s. A working solution of DCF-DA
Figure 1. Three different but synergistic testing principles are shown. was then prepared by adding 0.01 mL of stock to 10 mL of PBS. The
(A) The oxygen radical absorbance capacity (ORAC) assay is a chemical red blood cells were washed twice in PBS, resuspended in the DCF-
DA working solution, and incubated for 1 h at 37 °C. All samples,
test, in which interference with specific chemical reactions are measured.
except for the untreated control samples, were then exposed to 167
(B) The cell-based antioxidant protection in erythrocytes (CAP-e) assay mM H2O2 for a period of 45 min to induce severe oxidative stress.
reflects whether antioxidants can enter into and protect live cells from Samples were washed twice in PBS to remove the peroxide, transferred
oxidative damage. (C) The reactive oxygen species in polymorphonuclear to cold PBS, and stored on ice in preparation for immediate analysis
cells (ROS PMN) assay monitors the combined effect of a test product by flow cytometry. Intracellular levels of DCF-DA fluorescence
on an inflammatory cell type, and the data reflects a combination of at intensity in untreated versus H2O2-challenged cells were analyzed by
least three different mechanisms: (1) Antioxidants penetrate into the cell flow cytometry. Data was collected in quadruplicate for controls and
and neutralize free radicals, similar to the CAP-e assay; (2) Anti- in duplicate for each sample concentration. The mean fluorescence
inflammatory compounds mediate cell signaling at the cell surface, intensity (MFI) of red blood cells was compared between untreated,
reprogramming the PMN cell to a less inflammatory behavior, resulting in H2O2-treated, and cells pretreated with test products. A reduction in
MFI in samples pretreated with test products prior to challenge with
a reduction in formation of ROS; and (3) Pro-inflammatory compounds
H2O2 signified a reduction in oxidative damage mediated by the test
capable of supporting innate immune functions mediate a signal at the product. Experiments were repeated five times with similar results.
cell surface, resulting in an increase in the PMN cell function, thus
Assessment of Reactive Oxygen Species (ROS) Formation in
increasing the production of ROS. PMN Cells. The evaluation of anti-inflammatory action of test products
dimethyl sulfoxide (DMSO), Histopaque 1077, and Histopaque 1119. was done using freshly purified human PMN cells. The PMN were
Dichlorofluorescein diacetate (DCF-DA) was obtained from Molecular incubated at 37 °C, 5% CO2 for 90 min, either untreated or with test
Probes (Eugene, OR), a subdivision of Invitrogen (Carlsbad, CA). products over a range of dilutions from 0.01 to 0.0001 mg/mL. The
Natural Products. One animal-based, one microbial-based, one precursor dye DCF-DA was prepared by adding 0.18 mL of DMSO
plant-based, and one mixed natural product were obtained directly from and 0.02 mL of a 20% solution of Pluronic F-127 in DMSO to a 50 µg
the following individual suppliers: A colostrum whey-based extract, aliquot of DCF-DA and vortexing 3 times for 15 s. A working solution
Immunel (Sterling Technologies Inc., Brookings, SD), the high- of DCF-DA was then prepared by adding 0.01 mL stock to 10 mL
metabolite yeast culture-based product, EpiCor (Embria Health Sci- PBS. The PMN cells were washed twice in PBS, resuspended in the
ences, Ankeny, IA), freeze-dried Amazonian palm berry Acai (Eutherpe
¸ DCF-DA working solution, and incubated for 1 h at 37 °C. All samples,
oleracea Mart.) (K2A LLC, Provo, UT), and H.A. Joint Formula except for the negative control samples, were then exposed to 167 mM
(HAJF) (Purity Products, Plainview, NY). H2O2 for 45 min to induce severe oxidative stress. Samples were washed
Preparation of Natural Products Extracts. Natural products twice in PBS to remove the peroxide, transferred to cold RPMI, and
extracts were prepared in PBS in preparation for in vitro cell-based stored on ice. The DCF-DA fluorescence intensity was immediately
assays by adding 0.5 g of powder to 5 mL of PBS. This was mixed analyzed by flow cytometry, using a FacsCalibur flow cytometer
well by vortexing, placed on a rocker in the dark, and incubated at (Becton-Dickinson, San Jose, CA) and the CellQuest Pro (Becton-
room temperature for 1 h. The remaining solids were removed by Dickinson, San Jose, CA) and FlowJo (TreeStar, Ashland, OR) software.
centrifugation at 2400 rpm for 10 min followed by filtration of the Data was collected in triplicate for controls and in duplicate for each
supernatants using a 0.22 µm low protein binding cellulose acetate sample concentration. The MFI of PMN cells was compared between
syringe filter. The extracts were always used within 4 h of preparation, untreated, H2O2-treated, and cells pretreated with test products. A
as longer storage periods could result in oxidative breakdown of the reduction in MFI in samples pretreated with test products prior to
bioactive compounds that we were evaluating. challenge with H2O2 indicated a reduction in ROS production mediated
Purification of Peripheral Blood Polymorphonuclear Cells (PMN) by the test product. An increase in MFI in samples pretreated with test
and Red Blood Cells (RBC). Healthy human volunteers (n ) 12) products prior to challenge with H2O2 indicated an increase in ROS
between the ages of 20 and 60 years served as blood donors after production mediated by the test product. The experiments were repeated
informed consent, as approved by the Sky Lakes Medical Center three times with consistent results.
Institutional Review Board, was obtained. Peripheral venous blood As part of our initial testing, experiments were performed to reduce
samples were drawn into sodium heparin and layered onto a double- the contribution of pro-inflammatory signaling from the total results
gradient of Histopaque 1077 and 1119. The vials were centrifuged for in the ROS PMN assay. This was done by inhibiting cytoskeletal
25 min at 2400 rpm. The upper, PBMC-rich interface and the lower, movements involved in signaling and intracellular motility of vacuoles
PMN-rich interfaces were harvested using sterile transfer pipettes into involved in ROS formation. EpiCor was tested in two parallel sets of
new vials and washed twice in PBS without calcium or magnesium at tests, in which one set of duplicate tests was performed in culture
2400 rpm for 10 min. After the second wash, the PMN pellet was treated medium allowing full functionality of the PMN cells, and the parallel
with 4 mL of distilled H2O for 30 s to lyse red blood cells not removed set of duplicate tests were performed in culture medium, to which
by the gradient centrifugation. Immediately, 4 mL of 1.8% saline was sodium azide was added at a final concentration of 0.02%.
added to restore the physiological isotonic condition. Subsequently, 4 Oxygen Radical Absorbance Capacity (ORAC) Assay. Samples
mL of PBS was added, and the PMNs were centrifuged for 10 min at of Immunel and HAJF were shipped to Brunswick Laboratories, Norton,
2400 rpm. MA, an independent contract laboratory specializing in standardized
4. 8322 J. Agric. Food Chem., Vol. 56, No. 18, 2008 Honzel et al.
Table 3. Antioxidant Capacity of Test Products As Evaluated by ORAC Tests
ORAChydrophilic ORAClipophilic TAC NORAC HORAC SOD
natural product
(µmol TE/g) (µmol TE/g) (µmol TE/g) (µmol TE/g) (µmol GAE/g) (unit/g)
Immunel 18 NDa 18 0.59 1.72a NDa
OptiAcaib
¸ 997 30 1027 34 52 1614
HA Joint Formula 2219 139 2358 372 123 33
EpiCor 614 NDa 614 54 214 2200
a
ND ) not detected. b The antioxidant capacity for OptiAcai was previously published in ref 22.
¸
testing of natural products. The batch of EpiCor used for this project Immunel, known to provide a mild but significant effect in
was the same as the product used for in vitro testing of immunomodu- the CAP-e assay (27). As a plant-based extract, we used a
latory activity (24), but the results of the ORAC panel were not freeze-dried berry product OptiAcai from E. oleracea (acai),
¸ ¸
previously published. The ORAC values on acai were previously
¸ on which we have previously reported a strong ROS
published (22) and are reprinted here with the journal’s permission.
inhibitory effect in the PMN-based assay (22). In addition,
Data were obtained for all four natural products from ORAChydrophilic
testing using fluorescein as the fluorescent probe and 2,2′-azobis(2-
the joint health product HAJF was chosen. This product is a
amidinopropane) dihydrochloride as a peroxyl radical generator (11), blend of hyaluronic acid with hydroxytyrosol, a polyphenol
ORAClipophilic testing for lipid antioxidants capable of quenching peroxyl from olives. The yeast-based high-metabolite immunogen
free radicals (25), HORAC testing for antioxidants capable of quenching EpiCor was also included, based on its combination of high
hydroxyl free radicals (26), NORAC testing for antioxidants capable antioxidant content and immunomodulatory activity (24).
of quenching peroxynitrite, and SORAC testing for superoxide dismu- Comparison of CAP-e Data to the Effect on ROS Forma-
tase-like activity. tion in Human Polymorphonuclear (PMN) Cells. To illustrate
Statistical Analysis. Statistical analysis was performed using
the differences between the cell-based assays (Figure 1), we
Microsoft Excel. Statistical significance was tested using Student’s t
test with a p value of less than 0.05 indicating a significant difference
compared the effects of the four selected test products in
between data sets. both the CAP-e and the ROS PMN assays. For the ROS PMN
assay, freshly purified human PMN cells were used for
RESULTS assessment of modulatory effects on this inflammatory cell
type in vitro. Pretreatment of freshly isolated healthy human
Chemical Antioxidant Tests. The selected products were neutrophils with the four natural products before the induction
analyzed for total antioxidant capacity in a panel of ORAC of ROS by H2O2 treatment resulted in product-specific effects
assays (Table 3). The products were selected to include on ROS production.
products known to have various levels of protective activity
Immunel used for this study had a moderate ORAChydrophilic
in the CAP-e assay. As an example of an animal-based
value of 18 µmol TE/g, but antioxidants in Immunel were
product, we chose an extract from bovine colostrum whey,
available to live cells. Even more important, the data from
the CAP-e test suggested that the antioxidant compounds in
Immunel were retained within the cells, providing higher-
than-expected antioxidant protection at lower doses. In
addition, Immunel provided a mild but significant increase
in the formation of ROS by PMN cells, indicative of the
support Immunel provided to the innate immune system
(Figure 2).
Acai has a high ORAChydrophilic value of 997 µmol TE/g,
¸
and a significant amount of those antioxidants were able to
enter and protect live cells in the CAP-e assay (Figure 3).
In addition, acai inhibits ROS formation by PMN cells, even
¸
at extremely low doses. When comparing the CAP-e data to
the ROS PMN data for similar doses of acai, it became clear
¸
that the antioxidants alone could not account for the strong
anti-inflammatory effect of acai on PMN cells.
¸
HAJF, containing hydroxytyrosol from olives, had a very high
ORAChydrophilic of 2219 µmol TE/g. HAJF performed very well
in the CAP-e assay (Figure 4), indicating that the antioxidants
in HAJF were highly able to enter and protect live cells. HAJF
Figure 2. Comparison of the effects of the colostrum whey-based extract showed an inverted dose-response in the ROS PMN assay; that
Immunel in two different cell-based assays: The cell-based antioxidant is, the lower doses were more efficient at preventing ROS
protection of red blood cells (RBC) versus polymorphonuclear (PMN) cells. formation by PMN cells than higher doses. We interpret the
The RBC-based CAP-e assay shows that Immunel contains antioxidants inverse dose-response as the combined effect of hyaluronan,
able to enter and protect live cells from oxidative damage. The PMN- known to activate cells in the innate immune response, and the
based assay shows that Immunel triggers cellular signaling and ROS strong antioxidant capacity of hydroxytyrosol.
formation in PMN cells, as seen by the negative percent inhibition (i.e., EpiCor had a relatively high ORAChydrophilic value of 614
an increase in ROS formation). Thus, even though the PMN-based assay µmol TE/g, and a significant amount of those antioxidants
is not suitable for demonstrating the antioxidant capacity of the complex could enter and protect live cells, as shown by the CAP-e
product Immunel, the PMN-based assay demonstrates the immune assay (Figure 5). However, the PMN assay showed that
modulating property of Immunel. EpiCor at lower doses was able to induce a mild increase in
5. Antioxidant Methods for Foods and Natural Products J. Agric. Food Chem., Vol. 56, No. 18, 2008 8323
Figure 3. Comparison of the effects of the Amazonian palm berry acai ¸
in two different cell-based assays: The cell-based antioxidant protection
of red blood cells (RBC) versus polymorphonuclear (PMN) cells. The Figure 5. Comparison of the effects of EpiCor in three different cell-
freeze-dried acai contains antioxidants capable of entering and protecting
¸ based assays: The cell-based antioxidant protection of red blood cells
live cells in vitro. However, when a careful comparison across lower doses (RBC) versus polymorphonuclear (PMN) cells. The RBC-based CAP-e
of acai was performed in parallel using both the red blood cell (RBC)-
¸ assay showed that EpiCor contains antioxidants able to enter and protect
based CAP-e assay and the polymporphonuclear (PMN) cell-based assay, live cells in vitro, and the data demonstrate a clear and linear dose-
it was shown that the simple antioxidant content was not sufficient to response. The PMN-based assay was performed in two parallel manners:
explain the reduction in ROS formation in the PMN assay. We conclude (a) The exposure of the PMN cells to EpiCor was performed in culture
that the data from the PMN assay reflected strong anti-inflammatory medium, allowing the cells to react to signaling compounds present in
signaling of PMN cells by acai.¸ the EpiCor extract; (b) The exposure of the PMN cells to EpiCor was
performed in culture medium containing 0.05% sodium azide to block
cytoskeletal movements, thus abrogating many aspects of cellular signaling
and intracellular transport mechanisms involved in formation of reactive
oxygen species (ROS). The comparison between these two data sets on
PMN cells shows that when the PMN cells are able to perform cellular
signaling, the PMN cells respond to EpiCor by increasing their immune
reactivity and producing ROS. When cellular signaling is inhibited, some
antioxidant protection can be demonstrated in the PMN-based cell model.
The advantage of the CAP-e assay over the PMN-based assay is that
RBC do not produce ROS, neither as part of their immune function nor
as part of mitochondrial metabolism; and thus, the RBC provides a much
simpler and more conclusive cell-based model for antioxidant testing in
vitro.
was inhibited, the data showed that EpiCor also provided
protection from oxidative damage in the PMN assay, at least
as well as in the CAP-e assay.
DISCUSSION
Figure 4. Comparison of the effects of HAJF in two different cell- Both the dietary and nutraceutical industries are rapidly
based assays: The cell-based antioxidant protection of red blood cells moving beyond simple measures of nutrient content toward a
(RBC) versus polymorphonuclear (PMN) cells. HAJF showed a simple more complex understanding of the functionality of foods and
linear dose- response in the RBC-based CAP-e assay, indicating that supplements in biological systems. The industry is moving away
HA Joint Formula contains antioxidants capable of entering and from basing claims solely on methods based in analytical
protecting live cells from oxidative damage in vitro. However, an chemistry and embracing methods based on biological systems
opposite dose-response was seen in the PMN-based cellular assay, in vivo and in vitro. A continued forum for discussion is in
indicating opposing effects between different compounds in HAJF, high demand to support the creation of new standards for the
probably involving (1) antioxidants, (2) activating compounds, and (3) measurement of the effect of natural products in nonchemical
anti-inflammatory signaling compounds. testing systems.
With regard to antioxidants and products with anti-inflam-
ROS formation by PMN cells. Subsequently, EpiCor was matory properties, each available chemical tests has their
tested in the PMN assay using two parallel culture conditions: strengths and weaknesses because each is based upon detecting
(a) regular culture medium allowing the PMN cells full antioxidant interference with well-known, but limited, chemical
functionality; and (b) culture medium containing sodium reactions. Moving into cell-based testing systems in vitro, as
azide to abrogate cytoskeletal movements involved in cellular well as to testing in whole organisms in vivo, eliminates many
signaling and intracellular transport involved in ROS forma- of the limitations of the chemical interaction based assays. This
tion. By using the culture conditions in which the signaling has both positive and negative consequences. The positive aspect
6. 8324 J. Agric. Food Chem., Vol. 56, No. 18, 2008 Honzel et al.
is that the results are more relevant for biological systems. One the protection from hydroxyl radicals in the CAP-e assay. It is
of the down-sides is that a singular numerical value is no longer possible that EpiCor provides a better protection from peroxyl
easily obtainable. However, it is more important to apply free radicals, and work is in progress to adapt the CAP-e assay
numerous assays to gain a comprehensive understanding of a to include protection from peroxyl free radicals. However, the
product’s performance than to use a single numerical value to possibility also exists that EpiCor may be sending an anti-
describe a product. inflammatory signal to the PMN cells, thus serving to illustrate
The choice of immortalized cell lines for the purpose of all three aspects of PMN responses to compounds in natural
antioxidant testing is not a desirable approach. Some immortal- products, as outlined in Figure 1.
ized cell lines show altered functional responses to oxidative The CAP-e assay has use beyond in vitro evaluation of natural
stress, such as the human hepatocellular carcinoma line HepG2, products. The assay was applied to serum samples in a double-
which shows increased expression of catalase mRNA in blinded, placebo-controlled, clinical pilot study examining the
response to oxidative stress (28). Most immortalized cell lines, antioxidant uptake after consumption of the acai-rich juice
¸
including the HepG2 cell line, are hyperdiploid and perform MonaVie Active (34). Using the CAP-e assay in parallel to an
asymmetrical cell divisions, often rendering a proportion of the assay for serum lipid peroxidation, we showed that consumption
cells in culture dysfunctional and in various stages of cell death. of the juice resulted in a statistically significant improvement
The process of programmed cell death (apoptosis) leads to some of serum antioxidant status within 2 h after consumption. A
production of ROS by the cell’s mitochondria (29). A number single laboratory validation manuscript is currently being
of berry extracts have been found to induce apoptosis in several prepared to document the CAP-e method for broader use in the
types of tumor cell lines (30–32). If an antioxidant-rich natural natural products industry, including parallel testing with hy-
product reduces ROS production in such a cell model (33), the droxyl and peroxyl free radical generators, adaptation to an
interpretations are far from simple. One possible interpretation accelerated testing system (21), dose-response of a panel of
is that the product possibly protects tumor cells, which is not a antioxidant standards, and the effects of excipients on bioavail-
desirable conclusion. ability at the cellular level. Standards regarding assay proce-
In this study, we used three methods sequentially to examine dures, choice of cellular models, and the appropriate use and
antioxidant and anti-inflammatory properties of four selected interpretation of nonlinear dose-responses in cellular models
natural products. The products have very different ORAC values, are needed to ensure more consistency in reporting of natural
and yet the ORAC value alone did not do each product justice products in biological systems.
with regard to their overall biological effect. The product We have found the following sequential, multifacetted
Immunel had the lowest ORAC value among the four products strategy highly useful for the testing of natural products: (1)
tested but performed best in the CAP-e assay at low doses. In ORAC, (2) CAP-e, and (3) ROS PMN tests. When these tests
addition, Immunel supported the innate immune response by were used sequentially, a foundation of understanding was
providing a mild but significant enhancement of PMN function, generated, providing a more comprehensive understanding of
as seen by a negative percent inhibition of ROS formation antioxidant, anti-inflammatory, and immunomodulatory effects.
(Figure 2). The induction of ROS formation by PMN cells The four test products used in this paper serve to illustrate
should not be interpreted on its own as a damaging effect but different ways to look at complex antioxidant-containing natural
should be seen as an activation of one of our innate immune products. The data demonstrates the frequent nonlinearity of
defense mechanisms against bacterial invaders. It is interesting data in biological systems where the total observed effect
that while such an effect may be beneficial, the resulting risk depends on the interaction between compounds in the test
of oxidative damage may be buffered by antioxidants in the products and the nature of the cell model and cannot be analyzed
same complex natural product. by the more simple approaches used in analytical chemistry.
Both acai and HAJF showed a more linear dose-response in
¸
the CAP-e assay, but they each performed differently in the ABBREVIATIONS USED
ROS PMN assay. acai drastically reduced the PMN response
¸
across a wide dose range, and therefore, we expect that acai ¸ CAP, cell-based antioxidant protection assay; DCF-DA,
possesses strong anti-inflammatory properties in vivo. HAJF dichlorofluorescein diacetate; ORAC, oxygen radical absorbance
showed a more complex effect of PMN cells, indicating that capacity assay; PMN, polymorphonuclear cells; RBC, red blood
different compounds in the product support different aspects cells; ROS, reactive oxygen species.
of PMN cell biology. HAJF showed an opposite dose-response
in the PMN-based cellular assay (i.e., more effect at lower doses) ACKNOWLEDGMENT
possibly indicating opposing effects between different com- We thank Kelly M. Patterson, Lue Ann Zaske, and Aaron
pounds in HAJF, likely involving (1) antioxidants, (2) activating N. Hart for excellent technical assistance. We thank Sterling
compounds, and (3) anti-inflammatory signaling compounds, Technology Inc. (Brookings, SD), Purity Products (Plainview,
as outlined in Figure 1C. Using the EpiCor yeast extract in the NY), Embria Health Sciences (Ankeny, IA), and K2A LLC
ROS PMN assay under two parallel culture conditions, we were (Provo, UT) for permission to use data on their products.
able to show that EpiCor had immunomodulatory effects on
PMN cells. These effects prevented detection of EpiCor’s
antioxidant capacity in the PMN-cell based assay, unless LITERATURE CITED
cytoskeletal rearrangement was inhibited by sodium azide, thus
(1) Flora, S. J. Role of free radicals and antioxidants in health and
abrogating many aspects of cellular signaling and intracellular
disease. Cell. Mol. Biol. 2007, 53, 1–2.
transport necessary for ROS formation. Under those conditions, (2) Martınez, J. A. Mitochondrial oxidative stress and inflammation:
´
the PMN-based assay could demonstrate the antioxidant capacity An slalom to obesity and insulin resistance. J. Physiol. Biochem.
of EpiCor, in parallel to the RBC-based CAP-e assay. Interest- 2006, 62, 303–306.
ingly, at the highest dose, the inhibition of ROS formation by (3) Larbi, A.; Kempf, J.; Pawelec, G. Oxidative stress modulation
PMN cells was greater than what could be accounted for by and T cell activation. Exp. Gerontol. 2007, 42, 852–858.
7. Antioxidant Methods for Foods and Natural Products J. Agric. Food Chem., Vol. 56, No. 18, 2008 8325
(4) Victor, V. M.; Rocha, M. Targeting antioxidants to mitochondria: Amazonian palm berry, Euterpe oleraceae mart. (acai). J. Agric.
¸
A potential new therapeutic strategy for cardiovascular diseases. Food Chem. 2006, 54, 8604–8610.
Curr. Pharm. Des. 2007, 13, 845–863. (23) Patterson, K. M.; Yoon, I.; Jensen, G. S. Yeast culture has anti-
(5) Swerdlow, R. H. Treating neurodegeneration by modifying inflammatory effects and specifically activates NK cells. Comp.
mitochondria: Potential solutions to a “complex” problem. Anti- Immunol. Microbiol. Infect. Dis. In press (doi:10.1016/j.cim-
oxid. Redox Signaling 2007, 9, 1591–1603. id.2007.08.005).
(6) Klaunig, J. E.; Kamendulis, L. M. The role of oxidative stress in (24) Jensen, G. S.; Hart, A. N.; Schauss, A. G. An anti-inflammatory
carcinogenesis. Annu. ReV. Pharmacol. Toxicol. 2004, 44, 239– immunogen from yeast culture induces activation and alters
267. chemokine receptor expression on human natural killer cells and
(7) Federico, A.; Morgillo, F.; Tuccillo, C.; Ciardiello, F.; Loguercio, B lymphocytes in vitro. Nutr. Res. (N. Y.) 2007, 27, 327–335.
C. Chronic inflammation and oxidative stress in human carcino- (25) Huang, D.; Ou, B.; Hampsch-Woodill, M.; Flanagan, J. A.;
genesis. Int. J. Cancer 2007, 121, 2381–2386. Deemer, E. K. Development and validation of oxygen radical
(8) Cao, G.; Prior, R. L. Comparison of different analytical methods absorbance capacity assay for lipophilic antioxidants using
for assessing total antioxidant capacity of human serum. Clin. randomly methylated beta-cyclodextrin as the solubility enhancer.
Chem. 1998, 44, 1309–1315. J. Agric. Food Chem. 2002, 50, 1815–1821.
(9) Prior, R. L.; Wu, X.; Schaich, K. Standardized methods for (26) Ou, B.; Hampsch-Woodill, M.; Flanagan, J.; Deemer, E. K.; Prior,
the determination of antioxidant capacity and phenolics in foods R. L.; Huang, D. Novel fluorometric assay for hydroxyl radical
and dietary supplements. J. Agric. Food Chem. 2005, 53, 4290– prevention capacity using fluorescein as the probe. J. Agric. Food
4302. Chem. 2002, 50, 2772–2777.
(10) Ou, B.; Huang, D.; Hampsch-Woodill, M. Method for assaying (27) Patterson, K. M.; Carter, S. G.; Jensen, G. S. A novel fraction
the antioxidant capacity of a sample. U.S. Patent 7132296 B2. from bovine colostrum whey supports phagocytosis and natural
(11) Ou, B.; Hampsch-Woodill, M.; Prior, R. L. Development and killer cell activation while protecting cells from oxidative damage
validation of an improved oxygen radical absorbance capacity in vitro. Manuscript submitted for publication.
assay using fluorescein as the fluorescent probe. J. Agric. Food (28) Cuthbert, C.; Wang, Z.; Zhang, X.; Tam, S. P. Regulation of
Chem. 2001, 49, 4619–4626. human apolipoprotein A-I gene extression by gramoxone. J. Biol.
(12) Prior, R. L.; Hoang, H.; Gu, L.; Wu, X.; Bacchiocca, M.; Howard, Chem. 1997, 272, 14954–14960.
L.; Hampsch-Woodill, M.; Huang, D.; Ou, B.; Jacob, R. Assays (29) Jou, M. J.; Jou, S. B.; Chen, H. M.; Lin, C. H.; Peng, T. I. Critical
for hydrophilic and lipophilic antioxidant capacity (oxygen radical role of mitochondrial reactive oxygen species formation in visible
absorbance capacity (ORAC(FL))) of plasma and other biological laser irradiation-induced apoptosis in rat brain astrocytes (RBA-
and food samples. J. Agric. Food Chem. 2003, 51, 3273–3279. 1). J. Biomed. Sci. 2002, 9, 507–516.
(13) Prior, R. L.; Gu, L.; Wu, X.; Jacob, R. A.; Sotoudeh, G.; Kader, (30) Seeram, N. P.; Adams, L. S.; Zhang, Y.; Lee, R.; Sand, D.;
A. A.; Cook, R. A. Plasma antioxidant capacity changes following Scheuller, H. S.; Heber, D. Blackberry, black raspberry, blueberry,
a meal as a measure of the ability of a food to alter in vivo cranberry, red raspberry, and strawberry extracts inhibit growth
antioxidant status. J. Am. Coll. Nutr. 2007, 26, 170–181. and stimulate apoptosis of human cancer cells in vitro. J. Agric.
(14) Gurtovenko, A. A.; Anwar, J. Modulating the structure and Food Chem. 2006, 54, 9329–9339.
properties of cell membranes: The molecular mechanism of action (31) Mertens-Talcott, S. U.; Lee, J. H.; Percival, S. S.; Talcott, S. T.
of dimethyl sulfoxide. J. Phys. Chem. B 2007, 111, 10453–10460. Induction of cell death in Caco-2 human colon carcinoma cells
(15) Di, L.; Kerns, E. H. Biological assay challenges from compound by ellagic acid rich fractions from muscadine grapes (Vitis
solubility: Strategies for bioassay optimization. Drug DiscoVery rotundifolia). J. Agric. Food Chem. 2006, 54, 5336–5343.
Today 2006, 11, 446–451. (32) Bermudez-Soto, M. J.; Larrosa, M.; Garcia-Cantalejo, J. M.; Espin,
(16) Williams, A. C.; Barry, B. W. Penetration enhancers. AdV. Drug J. C.; Tomas-Barberan, F. A.; Garcia-Conesa, M. T. Up-regulation
DeliVery ReV. 2004, 56, 603–618. of tumor suppressor carcinoembryonic antigen-related cell adhe-
(17) Santos, N. C.; Figueira-Coelho, J.; Martins-Silva, J.; Saldanha, sion molecule 1 in human colon cancer Caco-2 cells following
C. Multidisciplinary utilization of dimethyl sulfoxide: Pharma- repetitive exposure to dietary levels of a polyphenol-rich choke-
cological, cellular, and molecular aspects. Biochem. Pharmacol. berry juice. J. Nutr. Biochem. 2006, 18, 259–271.
2003, 65, 1035–1041. (33) Wolfe, K. L.; Liu, R. H. Cellular antioxidant activity (CAA) assay
(18) Remick, D. G.; Villarete, L. Regulation of cytokine gene for assessing antioxidants, foods, and dietary supplements. J.
expression by reactive oxygen and reactive nitrogen intermediates. Agric. Food Chem. 2007, 55, 8896–8907.
J. Leukocyte Biol. 1996, 59, 471–475. (34) Jensen, G. S.; Patterson, K. M.; Barnes, J.; Carter, S. G.; Wu, X.;
(19) Gibson, G. E.; Huang, H. M. Mitochondrial enzymes and Scherwitz, L.; Beaman, R.; Endres, J. R.; Schauss, A. G. In Vitro
endoplasmic reticulum calcium stores as targets of oxidative stress and in vivo antioxidant and anti-inflammatory capacity of a juice
in neurodegenerative diseases. J. Bioenerg. Biomembr. 2004, 36, containing the Amazonian palm berry, acai (Euterpe oleracea
¸
335–340. Mart.). Manuscript submitted for publication.
(20) Buehler, P. W.; Alayash, A. I. Redox biology of blood revisited:
The role of red blood cells in maintaining circulatory reductive Received for review February 8, 2008. Revised manuscript received
capacity. Antioxid. Redox Signaling 2005, 7, 1755–1760. March 26, 2008. Accepted April 15, 2008. This work was sponsored in
(21) Jensen, G. S. Cell-based antioxidant protection assay. U.S. part by NIS Labs, which is an independent contract research laboratory
Provisional patent application No.60/985,166. specializing in natural products research, and in part by Merle West
(22) Schauss, A. G.; Wu, X.; Prior, R. L.; Ou, B.; Huang, D.; Owens, Center for Medical Research, a nonprofit research organization.
J.; Agarwal, A.; Jensen, G. S.; Hart, A. N.; Shanbrom, E.
Antioxidant capacity and other bioactivities of the freeze-dried JF800401D