POLI-Mix Functional Food Enhances Steady-State Bioenergetic Status Independently of Age: An Experimental Study Reza Rastmanesh, 1 Francesco Marotta, 2 Makoto Keiichi Kantah, 3 Ravinder Nagpal, 4 Aldo Lorenzetti, 2 Hiroshi Takadanohara, 3 Hiroshi Mashizume, 3 Riyichi Kobayashi, 3 and De Hua Chui 5 Abstract BALB/c mice were divided into young, middle-aged, and aged groups, and each group was given 3 weeks of oral treatments: (1) 1 mL of VBC1-99 (a mixture of 42 fruits and vegetables extracts) or (2) 1 mL of same amount of antioxidant vitamins as control. Steady-state hepatic adenosine triphosphate (ATP) was assessed by phosphorus-31 nuclear magnetic resonance ( 31 P-NMR) spectroscopy as: b-ATP/reference peak, inorganic phosphorus (Pi)/peak and b-ATP/Pi. As compared to untreated control, VBC1-99 significantly enhanced b-ATP/peak and b-ATP/Pi ratios ( p < 0.01) in all age groups and throughout the observation period ( p < 0.05) together with a significant decrease of Pi/ref peak ratio ( p < 0.05). However, this value in middle-aged and aged mice was comparable to antioxidant control mice. These NMR data demonstrate that VBC1-99 has a beneficial effect on hepatic energy metabolism, irrespective of age. Introduction A ged mammalian tissues show a decreased capacity to produce adenosine triphosphate (ATP) by oxidative phosphorylation due to dysfunctional mitochondria. The mitochondrial content of rat brain and liver is not reduced in aging, and the impairment of mitochondrial function follows a decreased rate of electron transfer by the selectively reduced complexes I–IV activities. One major mechanism is inhibition of the heme biosynthesis pathway in mitochon- dria, which causes a deficit of heme a. Mitochondrial oxidative dysfunction is accelerated by many common mi- cronutrient deficiencies. Indeed, minerals such as iron or zinc and several vitamin deficiencies, such as biotin or pan- tothenic acid, increase mitochondrial oxidative dysfunction and turnover through the aforementioned mechanism. Ames et al. 1 have suggested that an optimum intake of micronutrients could tune up metabolism and give a marked increase in health, particularly for the elderly, obese. Under conditions of increased ATP demand, liver mitochondria are able to increase ATP synthesis up to 2.8 times by switching more mitochondria from the resting state 4 to active state 3. However, aged hepatocytes, with a 30% complex I activity reduction, increase ATP production only up to 1.9 times. 2 Nonetheless, higher demands in younger subjects together with possible concurrent subtle micronutrients deficiencies may represent a further unsuspected health risk. Therefore, we examined steady-state energy status change in mouse liver of different age during 3 weeks treatment with VBC1-99 (a mixture of 42 extracts of fruits and vegetables), as compared to antioxidant-treated and untreated controls. As high-energy triphosphate compounds, we tested ATP using phosphorus- 31 nuclear magnetic resonance ( 31 P-NMR) spectroscopy, which allows noninvasive direct measurements of metabolite concentrations and tracing of metabolic fluxes in situ in ex- perimental animals and human livers. 3 Materials and Methods Animals A total of 120 healthy BALB/c mice raised in a controlled vivarium were divided into three separate age groups (40 mice each): Young (1 month), middle-aged (12 months), and aged (24 months). All mice were given water ad libitum and 1 Department of Clinical Nutrition & Dietetics, Shahid Beheshti University of Medical Sciences, National Nutrition and Food Technology Research Institute, Teheran, Iran. 2 ReGenera Res Group for Aging Intervention, Milan, Italy. 3 Suheiro Chem-Tech Center, Niigata, Japan. 4 Department of Biotechnology, JMIT Institute of Engineering and Technology, Radaur, Haryana, India. 5 Neuroscience Research Institute, Peking University, Beijing, China. REJUVENATION RESEARCH Volume 15, Number 2, 2012 ª Mary Ann Liebert, Inc. DOI: 10.1089/rej.2011.1271 161