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IMPACT OF AGENTS WITH POTENTIAL USE IN FUNCTIONAL FOODS ON BIOMARKERS FOR INDUCTION OF AGE RELATED DISEASES

Periodic Report Summary 3 - FUNCFOOD (IMPACT OF AGENTS WITH POTENTIAL USE IN FUNCTIONAL FOODS ON BIOMARKERS FOR INDUCTION OF AGE RELATED DISEASES)

Project Context and Objectives:
The overall aim of this project is to investigate the protective action of agents with potential use as functional food constituents with respect to cancer, diabetes and cardiovascular disease. The burden posed by these age related diseases on public health is enormous. Even a modest decrease of these ailments caused by consumption of appropriate functional foods translates into millions of lives saved. Based on animal experiments a number of agents have shown promise in preventing cancer as well as diabetes. However, proof of the utility of specific dietary ingredients as well as the utility of various biomarkers for determination of protection against these diseases in humans is incomplete.
The main objective for this project is to select food ingredients beneficial with respect to (i) genotoxicity/carcinogenicity as well as (ii) diabetes type 2 and associated cardiovascular complications (CVD). Some of these agents may prove suitable for inclusion in functional foods.

Project Results:
The strategy for this project was to have a tier approach starting with reviewing the literature for protective agents that can be a part of a human diet. After selection of 13 agents, in vitro methods were applied to screen for protection against cancer, diabetes and CVD using biomarkers that could be further used in animal models and in human intervention studies. The first in vitro experiments were performed with Chinese hamster cells and the best were selected for cytogenetic studies in vitro. The putative cancer protective compounds selected on the basis of screening were chlorophyllin (CHL), ellagic acid (EA), blueberry, Gentiana lutea extract (GL) and 6-gingerol (6-G).
Selected for test on cytogenetic parameters in Swiss albino mice were CHL, EA and BB and were found effective in reducing the frequency of micronuclei against methyl-nitroso-guanidine. The best protective agent against dimethylbenzanthracene was CHL whereas EA had no protective effects and BB had a weak protective effect.
Two human cohorts with intervention protocols were performed. Food including grilled sausages with high levels of PAHs, with and without combination of a protective agent were scheduled, followed by analysis of the incidence of Lymphocyte Micronuclei, Comet Tail Intensity, 8-oxo-dG (nM), analysis of CoQ10 and -tocopherol in blood and metabolites in urine. According to the findings it could be concluded that CHL, BB and tomato extract (TE) were protective supported by results of urinary metabolites. GL did not behave as an antioxidant, it affected mitotic spindle, induced malsegregation of chromosomes, apoptosis and necrosis. It should not be recommended for intake by healthy humans.
In the second human intervention study the protective effect of 5 selected agents in a cohort of totally 25 smokers were investigated. Samples of blood and urine were taken and distributed to the different laboratories for analysis. Each volunteer were investigated in terms of different life style factors such as general health status using standard clinical markers, questionnaire for eating habits, health profile, micronuclei in young erythrocytes (MN), 8-OxodG, haemoglobin adducts, urinary metabolites, endogenous levels of vitamin E & Q10. The best protective effect was found with BB, followed by astaxanthine (AX) and CHL. Limited support was obtained for protection by TE and GL. The biomarkers for diabetes in this cohort were to some extent successful, although the number of individuals were limited.
Diabetic dbb mice were used for studies of protection against diabetes, nephropathy and wound healing in vivo after treatments with CoQ, carnosine and AX. In comparison to wild type mice itt was found that diabetic mice at 8 months of age exhibited loss of sensation, hypoalgesia, decreased mechanical hyperalgesia, cold allodynia and decrease sciatic nerve conduction velocity. Daily CoQ treatment during 6 months improved these pathological conditions to a large extent. The histochemical study showed a significant reduction in the proportion of medium-sized neuron profiles. The study on mice was followed up by a study on humans investigating the preventive effect of 3 months treatment with carnosine and CoQ on the development of diabetes and its complications. Recruitment of 40 prediabetic and 40 type 2 diabetic patients without significant complications. The investigations so far performed showed promising results and included as biomarkers serum concentrations of coenzyme Q10 and vitamin E, hydrogen peroxide generation status. Reactive oxygen species (ROS) was determined by free oxygen radicals testing (FORT). A large number of relevant urinary and serum biomarkers, and waist index were determined using with established methods. Diabetic peripheral sensory neuropathy was assessed, and subjects with excessive consumption of alcohol, or treatment with B12, as well as those with a history of myocardial infarction and renal failure were excluded from the study.

Potential Impact:
Scientific impact of the FUNCFOOD project:
# The challenge posed by this project was to enhance the health benefit from diets beyond the traditional nutrients by means of addition of bioactive ingredients. This goal was achieved by the step-wise approach providing robust scientific evidence for selecting active ingredients appropriate for inclusion in functional foods.
# By starting with experimental models, the research also included mechanistic aspects, in as much as the impact of protective agents on carcinogen uptake and metabolism in humans was predicted, as well as the mode of action of bioactive ingredients on the prevention of age related diseases.
# The results achieved indicated that experimental in vitro and in vivo data could be predictable for humans in the sense that it provides rather qualitative than quantitative detection of preventing activity.
# Concerning the results on humans, several of the biomarkers showed evidence for detecting preventive mechanisms of background exposure, e.g. decreasing genotoxic risk, and risk for diabetes complications and CVD.
# Assessed by the biomarkers used in the humans in intervention, fresh and freezed-dried blueberry showed convincing evidence for being protective against cancer diabetes complications and CVD.
# Based on the overall results of over 300 biomarkers, astaxanthine, tomato extract and chlorophyllin showed probable evidence for being protective against cancer, whereas Gentiana extracts are suggested to have no or limited protective activity to any of the diseases.
# The results from the human intervention with all five antioxidants showed protective response at individual level to antioxidant intake only when background levels of chromosomal damage were indicated.
# In the cohort 100 individuals of different categories, several life-style factors were identified to influence genotoxic risk. Smoking, alcohol consumption and medical therapy were the most important for enhanced risk for cancer. The most important for protection were antioxidant intake and being a vegetarian.
# A tool was constructed for prediction of genotoxic risk which will be further developed and evaluated in forthcoming studies on human cohorts.


Socio-economic impact and social implications
# We here provided solid scientific data supporting that selected bioactive compounds in this project can be included in functional foods with potential health benefits for prevention and treatment of age related diseases.
# Identification of bioactive compounds for which dubious health claims have been made, can here on scientific grounds be neglected for being used in foods.
# We hereby verified that several of the biomarkers used in this project could monitor the beneficial effects of functional food constituents on human health.
# Data generated here can be explored in models for risk estimation as a basis for dissemination to the public in order to improve health status.

List of Websites:

funcfood.eu