Empirico-inductive and/or hypothetico-deductive methods in food science and nutrition research: which one to favor for a better global health?

Scientific research generally follows two main methods: empirico-inductive (EI), gathering scattered, real-world qualitative/quantitative data to elaborate holistic theories, and the hypothetico-deductive (HD) approach, testing the validity of hypothesized theory in specific conditions, generally according to reductionist methodologies or designs, with the risk of over simplifying the initial complexity empirically perceived in its holistic view. However, in current food and nutrition research, new hypotheses are often elaborated from reductionist data obtained with the HD approach, and aggregated to form (ultra)reductionist theories, with no application of EI observations, limiting the applicability of these hypotheses in real life. This trend and the application of the EI method are illustrated as regards with the global health issue through the examples of food classifications/scoring, clinical studies, the definition of a sustainable diet, the "matrix effect"-related hypothesis, the concept of healthy core metabolism, and obesity prevention within the perspective of social sciences. To be efficient for producing food and nutritional data appropriable by the society, it finally appears that not only both approaches are necessary, starting with the EI method then the HD one, but also a back and forth between the two, this being not always realized, potentially leading to confusion and misunderstanding in society.

Exclusive reductionism, chronic diseases and nutritional confusion: the degree of processing as a lever for improving public health.

Exclusive reductionism in nutritional science consists of viewing foods as only the sum of nutrients. This position paper argues that the extreme application of this paradigm since 1950 has greatly contributed to confusion about a healthy diet among consumers and to the development of chronic diseases worldwide. First, history of nutritional sciences in Western countries shows that by approximately 1850, laboratory research had mainly been conducted by reducing foods to nutrients that were interchangeable from one food to another. Second, descriptive and experimental studies show that the increased prevalence of chronic diseases mainly derive from ultra-processed foods. With such foods being representative of a final output in the degree of food processing, the relevance of reformulating food versus developing less unstructured processed foods is discussed. Third, the reductionist validation of food additives, randomized controlled trials, and food scoring is also questioned. Additionally, epidemiological studies that associate dietary patterns with the risk of chronic diseases and that aggregate approaches in nutrition, technology, food science and food scoring appear to be more adapted for nutritional recommendations in society. It is concluded that a complementary holistic perspective is needed to communicate to society about diet/food health potential and to efficiently prevent populations from chronic diseases.

Statistical mediation of the relationships between chronological age and lipoproteins by nonessential amino acids in healthy men.

Aging is a major risk factor for metabolic impairment that may lead to age-related diseases such as cardiovascular disease. Different mechanisms that may explain the interplay between aging and lipoproteins, and between aging and low-molecular-weight metabolites (LMWMs), in the metabolic dysregulation associated with age-related diseases have been described separately. Here, we statistically evaluated the possible mediation effects of LMWMs on the relationships between chronological age and lipoprotein concentrations in healthy men ranging from 19 to 75 years of age. Relative and absolute concentrations of LMWMs and lipoproteins, respectively, were assessed by nuclear magnetic resonance (NMR) spectroscopy. Multivariate linear regression and mediation analysis were conducted to explore the associations between age, lipoproteins and LMWMs. The statistical significance of the identified mediation effects was evaluated using the bootstrapping technique, and the identified mediation effects were validated on a publicly available dataset. Chronological age was statistically associated with five lipoprotein classes and subclasses. The mediation analysis showed that serine mediated 24.1% (95% CI: 22.9 - 24.7) of the effect of age on LDL-P, and glutamate mediated 17.9% (95% CI: 17.6 - 18.5) of the effect of age on large LDL-P. In the publicly available data, glutamate mediated the relationship between age and an NMR-derived surrogate of cholesterol. Our results suggest that the age-related increase in LDL particles may be mediated by a decrease in the nonessential amino acid glutamate. Future studies may contribute to a better understanding of the potential biological role of glutamate and LDL particles in aging mechanisms and age-related diseases.

Nutrition transition and chronic diseases in China (1990-2019): industrially processed and animal calories rather than nutrients and total calories as potential determinants of the health impact.

OBJECTIVE: To extend analyses of nutrition transition in developed countries to China within the framework of the 3Vs rule considering degree of processing starting with plant/animal calorie ratio (Rule 1), industrially processed foods (IPFs, Rule 2), and food diversity through nutrient intakes (Rule 3). DESIGN: Total and main food group (n 13) calorie intakes, percentages of animal and IPF calories, adequacy of the Dietary Reference Intake (DRI) and prevalence of chronic diseases were retrieved from scientific literature and international databases. SETTING: China, 1990­2019. PARTICIPANTS: Overall population. RESULTS: The total calorie intake decreased by 9 % over 30 years while the prevalence of chronic diseases substantially increased. Percentages of IPFs (Rule 1) and animal (Rule 2) calorie intake shifted from 9 to 30 % and 2 to 30 %, respectively. Meanwhile, the overall DRI adequacy (Rule 3) did not improve, with calcium and retinol deficiencies in 2019, and, although remaining above DRI, iron, copper, magnesium, and vitamins E, C and B1­B9 intakes regularly decreased. Notably, the prevalence of obesity increased five-fold, paralleling the exponential increase in IPF calorie intake. Both sources of calories were highly correlated with prevalence of main chronic diseases. CONCLUSIONS: Despite a slight decreased of total calorie consumption and small variations of adequacy with DRI, the farther the Chinese population moved away from the 3Vs rule during the 1990­2019 period, the more the prevalence of chronic diseases increased. Further analyses on foods' transitions will be better assessed when advocating sources/quality of calories (Rules 1/2), rather than only nutrient composition (Rule 3).

Precipitated sdLDL: An easy method to estimate LDL particle size.

BACKGROUND: LDL-C lowering is the main measure in cardiovascular disease prevention but a residual risk of ischemic events still remains. Alterations of lipoproteins, specially, increase in small dense LDL (sdLDL) particles are related to this risk. OBJECTIVE: To investigate the potential use of sdLDL cholesterol concentration (sdLDL-C) isolated by an easy precipitation method and to assess the impact of a set of clinical and biochemical variables determined by NMR on sdLDL concentration. METHODS: sdLDL-C and NMR lipid profile were performed in 85 men samples. Association among them was evaluated using Pearson coefficients (rxy ). A multivariate regression was performed to identify the influence of NMR variables on sdLDL-C. RESULTS: A strong association between sdLDL-C and LDLLDL-P (rxy  = 0.687) and with LDL-Z (rxy  = -0.603) was found. The multivariate regression explained a 56.8% in sdLDL-C variation (P = 8.77.10-12). BMI, ApoB, triglycerides, FFA, and LDL-Z showed a significant contribution. The most important ones were ApoB and LDL-Z; a 1nm increase (LDL-Z) leads to decrease 126 nmol/L in sdLDL-C. CONCLUSION: The association between sdLDL-C, LDL-Z, and LDL-P is clear. From a large number of variables, especially LDL-Z and apoB influence on sdLDL-C. Results show that the smaller the LDL size, the higher their cholesterol concentration. Therefore, sdLDL-C determination by using this easy method would be useful to risk stratification and to uncover cardiovascular residual risk.

Perspective: Reductionist Nutrition Research Has Meaning Only within the Framework of Holistic and Ethical Thinking.

Today, it seems that nutrition is in a state of great confusion, especially for the general public. For decades, some nutrients (e.g., cholesterol, saturated fats, sugars, gluten, salt) and food groups (e.g., dairy, cereals, meats) have been regularly denigrated. In this position paper, we hypothesize that such a state of confusion is mainly the result of the reductionist paradigm applied to nutrition research for more than a century, and by being pushed to its extreme, this perspective has led to accusations about some nutrients and foods. However, the real issue is about foods taken as a whole and therefore about their degree of processing, which affects both the food matrix and composition. Indeed, we eat whole foods, not nutrients. Therefore, the objectives of this article are to emphasize the need for more holistic approaches in nutrition to preserve our health, animal welfare, and planet. We propose to first redefine the food health potential on a holistic basis and then to show that reductionism and holism are interconnected approaches that should coexist. Then, we try to explain how extreme reductionism has been disconnected from reality and ethical considerations and has ultimately led to environmental degradation and loss of biodiversity, notably through very specific crops, and to an increased prevalence of chronic diseases. Furthermore, to address the confusion of the general public and to simplify nutritional messages, we propose 3 holistic golden rules based on scientific evidence to protect human health, animal welfare, and the environment (climate and biodiversity). Finally, we try to show how these 3 rules can be easily applied worldwide while respecting the environment, cultural traditions, and heritage.

Progesterone-associated arginine decline at luteal phase of menstrual cycle and associations with related amino acids and nuclear factor kB activation.

BACKGROUND/OBJECTIVES: Given their role in female reproduction, the effects of progesterone on arginine and related amino acids, polyamines and NF-κB p65 activation were studied across the menstrual cycle. METHODS: Arginine, ornithine and citrulline as well as putrescine, spermidine, spermine, and N-acetyl-putrescine were determined in plasma, NF-κB p65 activation in peripheral blood mononuclear cells and progesterone in serum of 28 women at early (T1) and late follicular (T2) and mid (T3) and late (T4) luteal phase. RESULTS: Arginine and related amino acids declined from T1 and T2 to T3 and T4, while progesterone increased. At T3, arginine, ornithine, and citrulline were inversely related with progesterone. Changes (ΔT3-T2) in arginine, ornithine, and citrulline were inversely related with changes (ΔT3-T2) in progesterone. Ornithine and citrulline were positively related with arginine, as were changes (ΔT3-T2) in ornithine and citrulline with changes (ΔT3-T2) in arginine. At T2, NF-κB p65 activation was positively related with arginine. Polyamines did not change and were not related to progesterone. All results described were significant at P < 0.001. CONCLUSIONS: This study for the first time provides data, at the plasma and PBMC level, supporting a proposed regulatory node of arginine and related amino acids, progesterone and NF-κB p65 at luteal phase of the menstrual cycle, aimed at successful preparation of pregnancy.

In vitro and in vivo antioxidant potential of milks, yoghurts, fermented milks and cheeses: a narrative review of evidence.

The antioxidant potential (AP) is an important nutritional property of foods, as increased oxidative stress is involved in most diet-related chronic diseases. In dairy products, the protein fraction contains antioxidant activity, especially casein. Other antioxidants include: antioxidant enzymes; lactoferrin; conjugated linoleic acid; coenzyme Q10; vitamins C, E, A and D3; equol; uric acid; carotenoids; and mineral activators of antioxidant enzymes. The AP of dairy products has been extensively studied in vitro, with few studies in animals and human subjects. Available in vivo studies greatly differ in their design and objectives. Overall, on a 100 g fresh weight-basis, AP of dairy products is close to that of grain-based foods and vegetable or fruit juices. Among dairy products, cheeses present the highest AP due to their higher protein content. AP of milk increases during digestion by up to 2·5 times because of released antioxidant peptides. AP of casein is linked to specific amino acids, whereas ß-lactoglobulin thiol groups play a major role in the AP of whey. Thermal treatments such as ultra-high temperature processing have no clear effect on the AP of milk. Raw fat-rich milks have higher AP than less fat-rich milk, because of lipophilic antioxidants. Probiotic yoghurts and fermented milks have higher AP than conventional yoghurt and milk because proteolysis by probiotics releases antioxidant peptides. Among the probiotics, Lactobacillus casei/acidophilus leads to the highest AP. The data are insufficient for cheese, but fermentation-based changes appear to make a positive impact on AP. In conclusion, AP might participate in the reported dairy product-protective effects against some chronic diseases.

Body mass index correlates with atherogenic lipoprotein profile even in nonobese, normoglycemic, and normolipidemic healthy men.

OBJECTIVE: To establish a relationship between body mass index (BMI), lipid, and lipoprotein parameters among nonobese, normoglycemic, and normolipidemic healthy men without any cardiovascular, metabolic, or chronic diseases. METHODS: A total of 297 healthy, nonsmoking males between 20 and 75 years were recruited. Exclusion criteria included familial hypercholesterolemia, any chronic diseases, and BMI ≥ 30 kg/m(2). Lipid and lipoprotein particles were determined by standard methods, with the use of ultracentrifugation and nuclear magnetic resonance (NMR). Cholesterol in remnant-like particles (RLPc) was also determined. RESULTS: These healthy volunteers were separated into two groups: normoweight (BMI > 19 kg/m(2) and <25 kg/m(2) [n = 143]) and overweight (BMI ≥ 25 kg/m(2) and <30 kg/m(2) [n = 154]). Overweight participants were older (P < .001) compared to normoweight. Both groups had low-density lipoprotein (LDL) cholesterol levels (<130 mg/dL) considered as desirable, and although both groups had plasma triglyceride levels within the nonpathological range, overweight participants presented with 30% higher triglyceride levels (P < .001) and 9% lower high-density lipoprotein cholesterol (P < .001) compared to normoweight individuals. Although LDL was comparable between groups, NMR analysis showed that overweight participants had 27% more total LDL particles due to a 16% decrease in large LDL (P < .001) and 70% increase in the smaller subclasses (P < .001). In overweight participants, NMR analysis also showed a 2-fold increase in large very low-density lipoprotein (P = .001), and 30% more medium very low-density lipoprotein particles (P = .020). Overweight participants also had 70% more intermediate-density lipoprotein particles (P = .010), a 30% decrease in large high-density lipoprotein particles (P < .001), and a 39% increase in RLPc levels (P = .005). Results were adjusted for age and fat intake. CONCLUSION: BMI correlates with a shift toward a more proatherogenic lipoprotein profile even in individuals whose lipid levels were not elevated.

Current food classifications in epidemiological studies do not enable solid nutritional recommendations for preventing diet-related chronic diseases: the impact of food processing.

To date, observational studies in nutrition have categorized foods into groups such as dairy, cereals, fruits, and vegetables. However, the strength of the association between food groups and chronic diseases is far from convincing. In most international expert surveys, risks are most commonly scored as probable, limited, or insufficient rather than convincing. In this position paper, we hypothesize that current food classifications based on botanical or animal origins can be improved to yield solid recommendations. We propose using a food classification that employs food processes to rank foods in epidemiological studies. Indeed, food health potential results from both nutrient density and food structure (i.e., the matrix effect), both of which can potentially be positively or negatively modified by processing. For example, cereal-based foods may be more or less refined, fractionated, and recombined with added salt, sugars, and fats, yielding a panoply of products with very different nutritional values. The same is true for other food groups. Finally, we propose that from a nutritional perspective, food processing will be an important issue to consider in the coming years, particularly in terms of strengthening the links between food and health and for proposing improved nutritional recommendations or actions.

Marked antioxidant effect of orange juice intake and its phytomicronutrients in a preliminary randomized cross-over trial on mild hypercholesterolemic men.

BACKGROUND & AIMS: Blond orange juice is the most consumed fruit juice in the world. It is a source of hesperidin, a bioavailable flavonoid reported to exhibit potential vascular protective actions. However, the specific impact on vascular function of Citrus phytomicronutrients, is unknown. For the first time, we investigated the effects of blond orange juice compared with a control beverage mimicking the composition of orange juice (including Vitamin C but no phytomicronutrients), on antioxidant markers, cardiovascular risk factors and endothelial function. METHODS: Twenty five male volunteers with two cardiovascular risk factors (age over 50 years and LDL-cholesterol between 130 and 190 mg/L) were enrolled in a randomized cross-over study. They received 3 times daily 200 mL of either blond orange juice or control beverage for 4 weeks, spaced by a 5-week wash-out. Endothelial function (flow mediated dilatation and plasma markers), oxidative status, lipid profile and inflammatory markers were assessed. RESULTS: Daily intakes of orange juice significantly led to a marked antioxidant effect which was correlated to hesperetin plasma levels and related with a decrease in reactive oxygen species. A tendency towards reduction of endothelial dysfunction and modest increase in plasma apoA-I concentration were also observed. This allows further experiments demonstrating the specific effect of phytomicronutrients from orange juice. CONCLUSIONS: These findings suggest that daily intake of nutritionally relevant dose of blond orange juice may contribute for a significant antioxidant effect through the phytochemicals contained in. Orange juice may be associated to other healthy foods to achieve a significant effect on the vascular function. This study is recorded in ClinicalTrials.com as NCT00539916.

Liposcale: a novel advanced lipoprotein test based on 2D diffusion-ordered 1H NMR spectroscopy.

Determination of lipoprotein particle size and number using advanced lipoprotein tests (ALTs) is of particular importance to improve cardiovascular risk prediction. Here we present the Liposcale test, a novel ALT based on 2D diffusion-ordered (1)H NMR spectroscopy. Our method uses diffusion coefficients to provide a direct measure of the mean particle sizes and numbers. Using 177 plasma samples from healthy individuals and the concentration of ApoB and ApoA from isolated lipoprotein fractions, our test showed a stronger correlation between the NMR-derived lipoprotein particle numbers and apolipoprotein concentrations than the LipoProfile(®) test commercialized by Liposcience. We also converted LDL particle numbers to ApoB equivalents (milligrams per deciliter) and our test yielded similar values of LDL-ApoB to the LipoProfile(®) test (absolute mean bias of 8.5 and 7.4 mg/dl, respectively). In addition, our HDL particle number values were more concordant with the calibrated values determined recently using ion mobility. Finally, principal component analysis distinguished type 2 diabetic patients with and without atherogenic dyslipidemia (AD) on a second cohort of 307 subjects characterized using the Liposcale test (area under the curve = 0.88) and showed concordant relationships between variables explaining AD. Altogether, our method provides reproducible and reliable characterization of lipoprotein particles and it is applicable to pathological states such as AD.

From a Reductionist to a Holistic Approach in Preventive Nutrition to Define New and More Ethical Paradigms.

This concept paper intends to define four new paradigms for improving nutrition research. First, the consequences of applying a reductionist versus a holistic approach to nutrition science will be discussed. The need for a more focused preventive nutrition approach, as opposed to a curative one, will then be presented on the basis of the 'healthy core metabolism' concept. This will lead us to propose a new classification of food products based on processing for future epidemiological studies. As a result of applying the holistic approach, health food potential will be redefined based on both food structure and nutrient density. These new paradigms should help define a more ethical preventive nutrition for humans to improve public recommendations while preserving the environment.

Proteomic analysis identifies cytoskeleton-interacting proteins as major downstream targets of altered folate status in the aorta of adult rat.

SCOPE: Mild folate deficiency and subsequently elevated plasma level of homocysteine are associated with an increased risk for vascular diseases in adults. Conversely, high intakes of folic acid (FA) may have beneficial effects on vascular function, presumably in part through homocysteine lowering. However, these effects have not yet been translated in terms of prevention or treatment of vascular pathologies. Besides, the complex biologic perturbation induced by variations of the folate supply is still not fully deciphered. We thus carried out a proteomic analysis of the aorta of adult rats after a dietary FA depletion or supplementation. METHODS AND RESULTS: Nine month-old rats were fed a FA-depleted, FA-supplemented or control diet for 8 weeks. Total proteins from adventitia-free aortas were separated by 2DE and differentially expressed proteins were identified by MS. FA depletion or supplementation resulted in significantly changed abundance of 29 spots (p < 0.05), of which 20 proteins were identified. Bioinformatic analysis revealed that most of these proteins are involved in cytoskeleton-related processes important to cell function/maintenance, assembly/organization, and movement. CONCLUSION: Our proteomic study supports that expression of proteins essential to vascular structure and, presumably, function is modulated by high intake as well as deprivation of FA.

Toward a new philosophy of preventive nutrition: from a reductionist to a holistic paradigm to improve nutritional recommendations.

The reductionist approach has been predominant to date in human nutrition research and has unraveled some of the fundamental mechanisms at the basis of food nutrients (e.g., those that involve deficiency diseases). In Western countries, along with progress in medicine and pharmacology, the reductionist approach helped to increase life expectancy. However, despite 40 y of research in nutrition, epidemics of obesity and diabetes are growing each year worldwide, both in developed and developing countries, leading to a decrease in healthy life years. Yet, interactions between nutrition-health relations cannot be modeled on the basis of a linear cause-effect relation between 1 food compound and 1 physiologic effect but rather from multicausal nonlinear relations. In other words, explaining the whole from the specific by a bottom-up reductionist approach has its limits. A top-down approach becomes necessary to investigate complex issues through a holistic view before addressing any specific question to explain the whole. However, it appears that both approaches are necessary and mutually reinforcing. In this review, Eastern and Western research perspectives are first presented, laying out bases for what could be the consequences of applying a reductionist versus holistic approach to research in nutrition vis-à-vis public health, environmental sustainability, breeding, biodiversity, food science and processing, and physiology for improving nutritional recommendations. Therefore, research that replaces reductionism with a more holistic approach will reveal global and efficient solutions to the problems encountered from the field to the plate. Preventive human nutrition can no longer be considered as "pharmacology" or foods as "drugs."

Does aging affect the immune status? A comparative analysis in 300 healthy volunteers from France, Austria and Spain.

BACKGROUND: As the European population is getting older, there is growing need in scientific data on how to achieve healthy and successful aging. A decline in immune function with age is unanimously supported by many epidemiological and clinical observations, with a decrease in T-cell mediated function encompassing a large part of this alteration. In the EU-funded VITAGE project, the effects of aging on biomarkers of immune status are being studied in three European countries. According to strict inclusion/exclusion criteria, a cohort of 300 healthy male non-smoking 20-75 years old volunteers were enrolled in France (n = 99), Spain (n = 100) and Austria (n = 101). In each country, the volunteers were classified as a function of age (one age group per decade). Biomarkers of immune status were determined including delayed-type hypersensitivity tests, measurement of lymphocyte surface markers, and serum determinations of interleukin-2, complement fractions and immunoglobulins. RESULTS: There were moderate differences in the biomarkers of immune status of the VITAGE study volunteers among the three European centres. The percentage of Natural Killer (NK) cells was 156% and 142% higher in Spain as compared to France and Austria, respectively (p < 0.0001), and this increase was observed at any age group above 30 years. Comparison between age-groups showed that in Spain, but not in France or Austria, older individuals had significantly a lower B lymphocyte distribution and conversely, a higher NK cell distribution. Moreover, the CD4/CD8 ratio was positively correlated with age in Austrian subjects (p < 0.0001). CONCLUSION: Our results provide evidence of an increased NK cell distribution in the elderly, especially in the Spanish population. NK cell status may predict morbidity and mortality in the elderly, emphasizing the importance of innate as well as adaptive immunity in ensuring healthy longevity and cancer resistance, possibly in link with the Mediterranean diet.

Immune status is more affected by age than by carotenoid depletion-repletion in healthy human subjects.

Prospective studies have indicated an age-related impairment of the immune response. Carotenoids have been hypothesised to enhance immune cell function. The aim of the present study was to describe the age-related effects and the impact of in vivo dietary carotenoid depletion and repletion on specific and non-specific immunity. A total of ninety-eight healthy male subjects (aged 20-75 years) received a carotenoid-depleted diet for 3 weeks and were then supplemented daily for 5 weeks with 30 mg ß-carotene, 15 mg lycopene and 9 mg lutein. Blood samples were collected at study entry, after depletion and supplementation, and biomarkers of immune status were determined. We found that serum IgA levels were positively correlated with ageing. Lymphocyte phenotyping indicated an increase with age in the memory T-helper cell subpopulation (CD4⁺CD45RO⁺) concomitantly with a decrease in naive T-helper cells (CD4⁺CD45RA⁺). A significant increase in the natural killer cells subpopulation and a small decrease in B lymphocytes were also observed, especially for the oldest volunteers. From ex vivo cell function exploration, a positive correlation was observed between age and IL-2 production of phytohaemagglutinin-stimulated lymphocytes. Neutrophils' bactericidal activity was significantly impaired with age (from 50 years) and was modulated by carotenoid status. An age effect was found on neutrophils' spontaneous migration but not on directed migration. Immune response in healthy human subjects is mostly affected by age rather than by dietary carotenoid depletion and repletion. Even in carefully selected healthy volunteers, some age-related immune changes occur predominantly from 50 years onwards. This immunosenescence could generate a loss in the immune system adjustment capacity.

Neutrophils are immune cells preferentially targeted by retinoic acid in elderly subjects.

BACKGROUND: The immune system gradually deteriorates with age and nutritional status is a major factor in immunosenescence. Of the many nutritional factors implicated in age-related immune dysfunction, vitamin A may be a good candidate, since vitamin A concentrations classically decrease during aging whereas it may possess important immunomodulatory properties via its active metabolites, the retinoic acids. This prompted us to investigate the immune response induced by retinoids in adults and elderly healthy subjects. Before and after oral supplementation with 13cis retinoic acid (0.5 mg/kg/day during 28 days), whole blood cells were phenotyped, and functions of peripheral blood mononuclear cells (PBMC) and polymorphonuclear cells (PMN) were investigated by flow cytometry and ELISA tests. RESULTS: In both young adults (n = 20, 25 ± 4 years) and older subjects (n = 20, 65 ± 4 years), retinoic acid supplementation had no effect on the distribution of leukocyte subpopulations or on the functions of PBMC (Il-2 and sIl-2R production, membrane expression of CD25). Concerning PMN, retinoic acid induced an increase in both spontaneous migration and cell surface expression of CD11b in the two different age populations, whereas bactericidal activity and phagocytosis remained unchanged. CONCLUSIONS: We demonstrated that retinoic acid induces the same intensity of immune response between adult and older subjects, and more specifically affects PMN functions, i.e. adhesion and migration, than PBMC functions.