Progress 10/01/05 to 09/30/10
Outputs
OUTPUTS: We identified corn distillers by-product as a major and inexpensive source of lutein and developed a method for lutein isolation from plant materials including aflatoxin-contaminated plant materials. A US Patent was applied for and issued. Two manuscripts are in prepation for submission to referred journals. A M.S thesis dissertation was defended by Naomi Sundalius in 2008. A monoclonal antibody was developed against the Bowman Birk inhibitor (BBI) for the determination of BBI levels in soy-containing food products. A publication on BBI was issued in 2008. Fenugreek as an ingredient in food for individuals with diabetes was studied and products containing fenugreek were developed. During 2010, three refereed journal publications were generated and results were disseminated through five presentations at the IFT annual meeting in Chicago. PARTICIPANTS: Jack N. Losso, Karen Mcdonough, and John W. Finley, LSU AgCenter. TARGET AUDIENCES: The targeted audience includes individuals with macular degeneration, diabetes, history of diabetes in their family, individual with angiogenesis-dependent diseases, and the general population. PROJECT MODIFICATIONS: Black seed oil, and auraptene have been added to the project.
Impacts
We identified corn distillers by-product as a major and inexpensive source of lutein. The isolated lutein was used to evaluate its efficacy against lipofuscin-catalyzed oxidation of retinal cells. We also investigated the interactions of blueberry anthocyanins and retinal cells in vitro and demonstrated that blueberry anthocyanins can protect retinal cells against lipofuscin-induced loss of viability of retinal cells. The mechanism of inhibition involved the inhibition of angiogenic biomarkers including VEGF. We also measured the levels of anthocyanins in a variety of berries grown across the US and in South America. Data will be used by Frito-Lay to develop lines of berry-rich food products. The efficacy of berry anthocyanins against low-grade inflammation was evaluated using human adipocytes as a model. Results show that berries can be effective in preventing low-grade inflammation stimulated by dietary free fatty acids or inflammatory lipopolysaccharides. The efficacy of resveratrol against hyperglycemia-induced inflammation and connexin degradation in retinal cells was investigated. Results indicate that resveratrol at low concentration can inhibit inflammation in the retinal cells by inhibiting the biomarkers of angiogenesis and protecting connexin. Research showed that the Bowman-Birk inhibitor (BBI) from soybean inhibits matrix metalloproteinase and prevents connexin degradation. We also investigated the efficacy of thymoquinone, a major bioactive compound in black seed oil, against the formation of advanced glycation end products which are commonly found in the eyes of people with diabetes and macular degeneration. Thymoquinone at physiologically acceptable concentration was effective in inhibiting the formation of advanced glycation end products ex vivo. The efficacy of black seed oil against the biomarkers of angiogenesis using a cancer cell line model is being pursued. The impact of the project has been significant in a number of areas including new scientific discoveries and new industry partnership opportunities. Ultimately, the impact on the consumer will be most important but is yet to be realized.
Publications
- Losso JN, Menelaou E, King JM. 2010. Isolation of aflatoxin-free lutein from aflatoxin-contaminated plants and plant products. US patent 7,671,242 B2.
- Losso JN, Bawadi HA, Chintalapathi M. 2011. Inhibition of the formation of advanced glycation ned products by thymoquinone. Food Chemistry. In press.
- Losso JN, Truax RE, Richard G. 2010. Trans-resveratrol inhibits hyperglycemia-induced inflammation and connexin down-regulation in retinal pigment epithelial cells. J. Agric. Food Chem. 58:8246-8252.
- Losso JN, Holliday DE, Richard, G. Karki N, Finley JW. 2010. The health benefits of Fenugreek. Cereal Foods World. 55: 236-241.
- Ganea GM, Fakayode SO, Losso JN, van Nostrum CF, Sabliov CM, Warner IM. 2010.Delivery of phytochemical thymoquinone using molecular micelle modified poly (D, L lactide-co- glycolide (PLGA) nanoparticles. Nanotechnology. 21:285104.
- Losso JN, Karki N, Jombai T, Mcdonough K, Grimm C, McMillin K, Finley JW, Enright FM. 2010. Inhibition of colon cancer stem cell biomarkers by black seed oil. Annual Meeting of the Institute of Food Technologists, Chicago, IL, July 17-21.
- Gao C, Mcdonough K, Finley JW, Losso JN. 2010. The effect of lutein and zeaxanthin against hyperglycemia and hypoxia on ARPE-cells. Annual Meeting of the Institute of Food Technologists, Chicago, IL, July 17-21.
- Karki N, JombaiT, Mcdonough K, Grimm C, McMillin K, Finley JW,Enright, FM, Losso JN. 2010. Black seed oil down-regulates breast cancer stem cell surface biomarkers more efficiently than fish oil or resveratrol. Annual Meeting of the Institute of Food Technologists, Chicago, IL, July 17-21.
- Soto AR, Finley JW, Losso JN, Reed TJ. 2010. Changes on the expression of three inflammatory biomarkers on human coronary arterial cells influenced by coffee phenolics of the Arabica and Robusta varieties. Annual Meeting of the Institute of Food Technologists, Chicago, IL, July 17-21.
- Soto AR, Finley JW, Losso JN, Richard GE. 2010. Effect of blueberry and black raspberry anthocyanins on the expression of PGE2-COX-p-p38 inflammatory pathway of human coronary arterial cells. Annual Meeting of the Institute of Food Technologists, Chicago, IL, July 17-21.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: In 2009, the following research activities, on pathological angiogenesis in general and angiogenesis in retinal cells in particular, were pursued. (1) A final US Patent application entitled "Isolation of aflatoxin-free lutein from aflatoxin-contaminated plants and plan products" was submitted to the US Patent office; a US patent will be issued in 2010. (2) Development of fenugreek-enriched wheat bread: We developed a fenugreek-enriched whole wheat bread for people with diabetes, a disease of excessive and insufficient angiogenesis. A clinical study conducted at Pennington Biomedical Research Center showed that fenugreek-enriched bread improves insulin sensitivity in diabetic individuals. (3) Sulforaphane and phenethyl isothiocyanate were investigated as inhibitors of inflammation that leads to pathological angiogenesis. Results were published in the spring of 2009. (4) Ceramides inhibition of pathological angiogenesis: We investigated and demonstrated that ceramides from oysters inhibit angiogenesis in endothelial cells and the proliferation of hormone-dependent and independent breast cancer cells in vitro. (5) Potential of resveratrol against angiogenesis in retinal cells: We investigated the interactions of resveratrol and human retinal cells under normal and hyperglycemic conditions. The results of our study indicate that resveratrol at low concentration can inhibit inflammation in the retinal cells. (6) Development of a monoclonal antibody against the Bowman-Birk inhibitor (BBI) for the analysis of BBI, as anti-angiogenic compound in food products. Work is in progress. (7) Development of marine collagen peptides as inhibitors of angiogenesis. Using our patented process (US Patent 7,109,300 B2) for marine collagen isolation, we have been working on developing collagen peptides from black drum or sheepshead collagen for use in biomedical and beverage applications. Work is till in progress for this effort. (8) Identification and Evaluation of Berries with Superior Functional and Processing Properties. Funded by Pepsi-Cola through Frito-Lay we examined several varieties of berries harvested from across the US and Chile for total anthocyanins, anthocyanins profile, and antioxidant activity. Because of the proprietary nature of the research, only progress and final reports were required. These reports have been submitted to Frito-Lay. As a result, Frito-Lay asked us to assist in the analyses of anthocyanins from some of their products after the conclusion of the projects. (9) Development of black seed oil as inhibitor of pathological angiogenesis. We have identified black seed oil as dietary oil that has the potential to inhibit angiogenesis in several angiogenesis-dependent diseases including macular degeneration and cancer. While our research is still on-going, several manuscripts have been drafted for submission and in 2010 this will be a major focus of our research publications. PARTICIPANTS: Robert Truax, formely with LSU AgCenter John Finley, Head and Professor of Food Science Roy Martin, Human Ecology and LSU AgCenter Frank Greenway, Pennington Biomedical Research Center Jennifer Rood, Pennington Biomedical Research Center Y. Yu, Pennington Biomedical Research Center Madhavi Chintalapathi, graduate student Food Science Darryl Holliday, graduate student Food Science TARGET AUDIENCES: The target audience for our research is widespread: individuals with macular degeneration, diabetes complications, and individuals with other angiogenic diseases. PROJECT MODIFICATIONS: We have added "black seed oil, sulforaphane, phenethyl isothiocyanate, and vertebrate collagen as part of the list of bioactive compounds to be investigated.
Impacts
The impact of our work was significant in 2009. (1) On January 26-28 representatives from the Louisiana oyster industry invited the PI to travel to Washington, DC with a delegation of oyster industry representatives to meet with US Senators from Louisiana, Mississippi, Florida, Alabama, and Texas or their staff members. I was invited to give a brief presentation on my research on oysters and their potential health benefits associated with ceramides. On January 28, 2009 I was a guest speaker at the NOAA Sea Grant National Library in Silver Spring, MD. The title of my presentation was "Compounds in oysters are effective in preventing cancer cell growth". (2) Another impact of work on angiogenesis was illustrated by the invitation that I received from the CEO of the Angiogenesis Foundation to speak at the Judah Folkman International Conference on Angiogenesis on October 12-13, Cambridge, MA. I was one of the 16 speakers (14 medical doctors, me, and one cancer survivor) invited to give a 30 minutes lecture on their research activity. The topic of my presentation was "Angioprevention discovery: land and sea screening". After my presentation I was offered to be on the faculty advisory board of the foundation and was asked to assist in organizing the 2010 International Conference section on dietary angiopreventive compounds. I am also working with the foundation on a research proposal on dietary anti-angiogenic factors. (3) The impact of my work on Angiogenesis was also realized when I received an invitation from the CEO (Rexford Titus) of a healthcare insurance company, Pro-HealthCare, with headquarters in Pewaukee, WI to speak to medical doctors and insurance executives about angiogenesis and chronic degenerative diseases. I gave a presentation entitled "Angioprevention with dietary factors" on November 18 to medical doctors and healthcare executives. (4) The impact of fenugreek has been good and we are still making progress to have a company acquire our technology to make fenugreek bread. (5) The impact of collagen is an ongoing effort to have Omega Protein, a major menhaden processing company based in Houston, TX adopt our patented process for the isolation of collagen from menhaden waste.
Publications
- Losso JN, Holliday DL, Finley JW, Martin RJ, Rood JC, Yu Y, Greenway FL. 2009. Fenugreek bread: a treatment for diabetes mellitus. J. Med. Food 12:1046-1049
- Chintalapathi M, Truax RE, Stout R, Portier RJ, Losso JN. 2009. In vitro and in vivo antiangiogenic activities and inhibition of hormone dependent and independent breast cancer cells by ceramide amethymainoethylphosphonate. J. Agric. Food Chem. 57:5201-5210
- Tao Dong, John W. Finley, Jack N. Losso. 2009. The anti-inflammation effect of muscadine juice on human coronary arterial smooth muscle and endothelial cells. Presented at the Annual meeting of the Institute of Food Technologists, Anaheim, CA, June 6-9,2009
- Tao Dong, John W. Finley, Jack N. Losso, Robert E. Truax, Gerald I. Richard, Mark Dreher. 2009. The anti-inflammatory effect of pomegranate juice on human coronary arterial smooth muscle and endothelial cells. Presented at the Annual meeting of the Institute of Food Technologists, Anaheim, CA, June 6-9,2009
- Losso JN, Truax, RE. 2009. Comparative inhibitory activities of sulforaphane and phenethyl isothiocyanate against leukemia resistant CEM/C2 cancer cells. J. Functional Foods 1:229-235
- Adriana Soto, John W. Finley, Jack N. Losso, Gerald I. Richard, Robert E. Truax. 2009. Effect of blueberry (Vaccinium spp) anthocyanins on the expression of inflammation biomarkers on human coronary arterial smooth muscle cells. Presented at the Annual meeting of the Institute of Food Technologists, Anaheim, CA, June 6-9,2009
- Adriana Soto, John W. Finley, Jack N. Losso, Gerald I. Richard, Robert E. Truax. 2009. Changes on the expression of inflammation biomarkers on human coronary arterial smooth muscle and endothelial cells influenced by black raspberry (Rubus occidentalis) anthocyanins. Presented at the Annual meeting of the Institute of Food Technologists, Anaheim, CA, June 6-9,2009
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: From this project, one M.S. thesis was written and defended in December 2008 covering the potential health benefits of anthocyanins from blueberries against lipofuscin (A2E)-induced degradation of retinal cells through accumulation of low-grade inflammation and angiogenic biomarkers. From this project, work has been done, food products made, and a manuscript accepted for publication to demonstrate the health benefits of consuming fenugreek-enriched food products such as fenugreek bread, fenugreek tortilla, and fenugreek cookies. From this project, we also published a paper that promotes the use of soybean protein known as the Bowman-Birk inhibitor (BBI) as an important functional food that helps maintain cells in the body in communication even under disease condition so that treatment can be effective. From this project, we have also shown that resveratrol, commonly found in red wine, is a strong inhibitor of oxidative stress in retinal cells and prevention of hyperglycemia-induced retinal cell degradation that leads to macular degeneration in diabetic individuals. The work on resveratrol is still in progress and a manuscript has been prepared for submission. We have also concluded that lutein in combination with phosphatidylglycerol can significantly reduce the damaging effects of lipofuscin in retinal cells. We have isolated lutein from distillers grains, a byproduct of ethanol production from corn, and demonstrated that lutein is stable during corn fermentation for ethanol production and the isolated lutein can protect retinal cells against lipofuscin and peroxide-catalyzed oxidation in retinal pigment epithelial cells. The significance of our findings have been shared with readers around the US and world and corn processors in Louisiana. PARTICIPANTS: Jack N. Losso, LSU AgCenter; Robert E. Truax, LSU AgCenter; Naomi M Sundalius, LSU Food Acience; John W. Finley, LSU AgCenter. TARGET AUDIENCES: The targeted audience includes individuals with macular degeneration, diabetes, history of diabetes in their family, and the general population. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
As the continuation of our research on the effects of blueberry anthocyanins on A2E-induced retinal cell degradation (which was partially reported in 2007), we have determined that blueberry anthocyanins when present in retinal cells can delay retinal cell degradation and reduce the generation of low-grade inflammation such as IL-6 and IL-8 and angiogenic biomarkers such as VEGF. We have also shown that whole fenugreek-enriched bread can control hyperglycemia which is a stimulator of macular degeneration. Research on resveratrol has indicated that this compound is a good inhibitor of low-grade inflammation in the retinal cells. Our work on BBI protein suggests that this protein promotes cell to cell communication which is a critical factor to prevent the onset and progression of macular degeneration and several other chronic degenerative diseases. The impact of our studies is as follows. Anthocyanins in blueberries prevent oxidants in the eyes from damaging retinal cells through continuous inhibition of oxidative stress. Oxidative stress is known to generate radicals which, if not inactivated, progressively destroy the ability of retinal cells to remain together and help individual see things clearly. Information on the health benefits of blueberry anthocyanins is already assisting blueberry growers, processors, and consumers in producing more blueberries, processing blueberries under the best conditions that retain the health attributes, and consuming blueberries to remain healthy and maintain good vision. The information on fenugreek is already being discussed with fenugreek producers to launch fenugreek-enriched cereal and baked products for individuals with diabetes, individuals susceptible to diabetes, and the general population to prevent the risk of diabetes.
Publications
- Naomi M. Sundalius.Examination of blueberry anthocyanins in prevention of age-related macular degeneration through retinal pigment epithelial cell culture. M.S. Thesis, Louisiana State University, 2008.
- Losso, JN. 2008. The biochemical and functional food properties of the Bowman-Birk inhibitor. Critical Review in Food Science and Nutrition. 48:94-118.
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: From this project, one US patent application has been filed. US Patent Application 20070142678: Isolation of aflatoxin-free lutein from aflatoxin-contaminated plants and plant products. June 21, 2007. Also from this project, a scientific presentation was made at the Annual meeting of the American Chemical Society, Aug 19-23, 2007, Boston, MA.; Annual Meeting of the Institute of Food Technologists, July 28-August 1, 2007, Chicago, IL. The significance of our findings were shared with participants at the meeting.
PARTICIPANTS: Robert Truax, AgCenter Biotech Lab, collaborator; Naomi Sundalius, Graduate student; USDA-Blueberry Council,funding agency; Louisiana Soybean and Grain Promotion and Research Board, previous funding
TARGET AUDIENCES: academics and general population
Impacts
We have determined the following: 1. Anthocyanins in blueberry can protect retinal cells from oxidative stress induced by lipofuscin (A2E). 2. Lutein prevents the degradation of gap junctional communication in retinal cells. 3. Phosphatidylglycerol is a powerful inhibitor of oxidative stress in retinal cells. The impact of our studies is as follows. Anthocyanins in blueberries are important health enhancing bioactive compounds that act as antioxidants and prevent radical formation associated with oxidative stress. The inhibition of oxidative stress has been linked to enhanced health. Information on the health enhancing effects of blueberries can assist growers, processors, and consumers in marketing blueberries, processing them to retain quality, and consuming these berries for better vision. The information on phosphatidylglycerol will also assist food processors and formulators in incorporating this emulsifier into food products. Similarly consumers will benefit by
consuming health enhancing phosphatidylglycerol for healthy vision.
Publications
- ABSTRACTS 1. Losso, J.N. and Truax, R.E. 2007. Protective effect of lutein against lipofuscin and peroxide-catalyzed oxidation, proliferation, and gap jucntional communicationdegradation of retinal pigment epithelial cells. Annual Meeting of the Institute of Food Technologists, Chicago, IL July.
- 2. Losso, J.N. 2007. Interactions of functional foods and age-related macular degeneration. Annual Meeting of American Chemical Society, Boston, Ma.
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Progress 01/01/06 to 12/31/06
Outputs
We are investigating lutein, blueberry anthocyanins and phosphatidylglycerol as functional food inhibitors of lipofuscin , N-retinyl-N-retinylidene (A2E) accumulation in the RPE cells. 1. Protective effect of lutein against lipofuscin and peroxide-catalyzed oxidation, proliferation and gap junctional communication degradation of retinal pigment epithelial cells - The interactions of A2E and retinal pigment epithelial cells are being studied in vitro in absence or presence of lutein (100 micromoles) before or after the cells have been exposed to 100 umol/L of A2E, illumination under a fluorescent light for 20 min, exposure to 100 umol/L of peroxide, and 6 days incubation at 5 percent CO2 and 37 degrees C. In the absence of lutein, A2E caused retinal cell degradation as demonstrated by the increase in the levels of angiogenic proteins such pigment epithelium derived factor (PEDF), vascular endothelial growth factor (VEGF), and loss of gap junctional communications.
Lutein at 100 umol/L protected RPE cells better when cells were pre-incubated with lutein for 24 h followed by addition of A2E and peroxide. In the later case, cell viability was enhanced with time and cells were 70-80 percent confluent within 72 h. The levels of PEDF and connexin 43 remained unchanged in cells treated with lutein, A2E, illumination and peroxide, whereas cells treated with A2E and peroxide lost significant levels of PEDF and connexin 43. 2. Synergistic Effects of Blueberry Phytochemicals Against Age-Related Macular Degeneration - The objective of this research is to examine whether a mixture of blueberry phytochemicals can prevent the accumulation of A2E in RPE cells in vitro and stabilize the anti-angiogenic protein pigment epithelium derived factor (PEDF) and prevent the increased expression in the RPE cells of angiogenic factors VEGF, IGF-1, TNF-a, IL-8, MCP-1, and MAP kinase in vitro. Blueberry anthocyanins or blueberry extracts were incubated with RPE in absence
or presence of A2E for 6 days. Briefly, A2E caused RPE cells degradation as demonstrated by cell morphology. A2E interaction with RPE was associated with an increase in the levels of angiogenic proteins VEGF, PlGF, IL-8, and MCP-1. Interactions of blueberry powder at 500 ug/ml or blueberry anthocyanins at 250 ug/ml time-dependently reduced the levels of angiogenic proteins mentioned above. Within 6 days of incubation, RPE cells recovered (similar results were obtained with blueberry anthocyanins) and the levels of the angiogenic proteins decreased significantly. 3. Biosynthesis of phosphatidylglycerol - Phosphatidylglycerol holds promise as a potent inhibitor of ARMD. However, availability and costs limit the use of this bioactive compound as functional food and emulsifier. We are using egg yolk and soybean as sources of phosphatidylcholine from which phosphatidylglycerol can be biosynthesized. We succeeded in converting phosphatidylcholine to phosphatidylglycerol using phospholipase
D but the conversion rate is slow (more than 12 hours). Work is in progress to optimize the reaction and conversion rates.
Impacts
Bioactive compounds such as lutein and anthocyanins when consumed in adequate amounts may help reduce the incidence and progression of age-related macular degeneration.
Publications
- Menalaou, E. Khachatryan A. Losso, J.N. Cavalier, M., Labonte, D. 2006. Lutein content in sweet potato leaves. Hortscience 41(5): 1-3.
- Losso, J. 2007. History and Scope of Functional Foods in the USA. Chapter 1. In Anti-angiogenic functional and medicinal foods. J. Losso, D. Bagchi, and F. Shahidi (eds.). Taylor and Francis.
- Losso, J. 2007. Screening functional foods as inhibitors of angiogenesis biomarkers. In Anti-angiogenic functional and medicinal foods. J. Losso, D. Bagchi, and F. Shahidi (eds.). Taylor and Francis.
- Bagchi, M., Zafra, S., Losso, J., Sen, C.K., Roy, S.,Hazra, S., Bagchi, D. 2007. Role of edible anthocyanins in angiogenesis. In Anti-angiogenic functional and medicinal foods. J. Losso, D. Bagchi, and F. Shahidi (eds.). Taylor and Francis.
- Losso, J. 2007. Development and delivery of anti-angiogenic functional foods. In Anti-angiogenic functional and medicinal foods. J. Losso, D. Bagchi, and F. Shahidi (eds.). Taylor and Francis.
- US Patent Losso, J.M., Ogawa. M., Portier, R.J., Schexnayder, M.A. 2006. Extraction of collagen from calcified tissues. US Patent No. 7,109,300,B2
- Losso, J.N., King, J.M., Menelaou, E. 2005. Isolation of flatoxin-free lutein from aflatoxin contaminated plant and plant products. US Patent Applied.
- Book Edited Anti-angiogenic functional and medicinal foods. J. Losso, D. Bagchi, and F. Shahidi (eds.). Taylor and Francis. 2007.
- Book Chapters Losso, J.N. 2007. Using marine by-products in pharmaceutical, medical, and cosmetic products. in Maximising the value of marine by-products. F. Shahidi (ed.) pp. 450-456.
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