Effects of Diacetyl Exposure in the Metabolism of Male and Female Mice

Metabolism and Bioenergetics 433 Effects of Diacetyl Exposure in the Metabolism of Male and Female Mice Leticia Dias Lima Jedlicka1, Juciara Costa Silva2, Aleksandro Martins Balbino1, Giuseppe Bruno Neto1, Heron Dominguez Torres Silva1, Danielle Zildeana Souza Furtado1, Fernanda de Barros Correia Cavalcanti2, Karin Marrie van de Heijden2, Carlos Alberto Avellaneda Penatti2, Etelvino José Henriques Bechara3, and Nilson Antonio Assuncao1 1 UNIFESP, Brazil, 2UNINOVE, Brazil, 3USP, Brazil The search for safe and high quality food has become an increasing concern in recent decades. This phenomenon is due to the increased consumption of both processed and artificially preserved foods. The food industry has increasingly improved organoleptic characteristics and storage of food using food additives. Diacetyl (2,3-butanedione) is a flavoring that imparts a buttery flavor to foods such as microwave popcorn, cookies, wine, beer, hard liquor and dairy products. In this work, we investigated the effect of oral intake of 2,3-butanedione in C57/Bl mice (male and female). We performed a target metabolomics assay using ultra performance liquid chromatography–triple-quadrupole (UPLC-MS/MS) for the determination and quantification of plasmatic metabolites. We observed alterations in the metabolites present in some important metabolic routes such as the urea and tricarboxylic acid (TCA) cycles. Peroxynitrite plasmatic levels were also evaluated by colorimetric method and final activity of superoxide dismutase (SOD) evaluated by enzymatic method. Additionally, mouse behavior was evaluated. Most of the assays showed differences between control and treatment groups and between genders, which may indicate the involvement of sex hormones in the regulation of a normal metabolic profile and the implication of sex differences in metabolite disease response.

doi: 10.1016/j.freeradbiomed.2016.10.474 434 Absence of Functional Sirtuin 3 Alters Fatty Acid and Glucose Metabolism Following 4-OH-PCB11 Exposure Sinthia Jahan1, Gwendolyn S. Carter1, Kimberly J. Krager1, Xueshu Li2, Hans-Joachim Lehmler2, and Nukhet Aykin-Burns1 1 University of Arkansas for Medical Sciences, Little Rock, USA, 2 The University of Iowa, Iowa City, USA Sirtuin3 (Sirt3) is the major deacetylase in mitochondria. It has been determined that SIRT3 null mice have deficiencies in ATP production and demonstrate a susceptibility to develop metabolic syndrome. Polychlorinated biphenyls (PCBs) are organic pollutants that accumulate in adipose tissue, and have been shown to disturb metabolism. They have been suggested to contribute to metabolic disease states such as diabetes and obesity. Our goal is to examine the effects of 4-hydroxy-3,3’-dichlorobiphenyl (4OHPCB11), a major PCB 11 metabolite, on fatty acid and glucose metabolism using embryonic fibroblasts (MEF) isolated from SIRT3 wild type and SIRT3 null mice. RT² Profiler™ PCR Array for fatty acid metabolism exhibited a dose dependent up-regulation of Acot12, Acsbg2, Acsm2, Fabp1, Oxct2a, Gk2, Hmgs2, LpI,

SLC27A5 and Acsl1 genes in Sirt3–/– MEFs compared to Sirt3+/+ MEFs following 24 h treatment with 0.1, 1, and 3 μM 4OH-PCB11. PCR Array for glucose metabolism also demonstrated upregulation of G6pc, Pdk4, and Prps1L1 in both Sirt3–/–and Sirt3+/+MEFs upon 3 μM 4OH-PCB11 exposure, however fold increases in the expression of these genes were more pronounced in the knockout background. On the other hand, the expression of Pygl gene was down-regulated in both Sirt3–/–and Sirt3+/+MEFs at comparable levels. Our future studies will investigate the enzymatic activities of proteins encoded by these genes as well as utilize a proteomics approach to determine if they are specific Sirt3 targets during PCB induced cellular stress. Supported by NIGMS P20- GM109005, NIEHS R15 ES022781, and P42 ES013661.

doi: 10.1016/j.freeradbiomed.2016.10.475 435 Metabolomic-Based Amino Acid and Biogenic Amine Plasmatic Profile of Brazilian Cri-du-Chat Patients Etelvino Bechara1, Danielle Furtado2, Fernando Leite2, Cleber Barreto2, Elisangela Silva3, Heron Silva2, and Nilson Assunção2 1 University of São Paulo, Brazil, 2Federal University of São Paulo, Brazil, 3AC Camargo Cancer Research, Brazil Cri-du-chat syndrome (CDCS) is a rare innate disease attributed to chromosome 5p deletion characterized by a cat-like cry, craniofacial malformation, and altered behavior of affected children. Here, UHPLC/MS and chemometrics were employed to analyze blood samples withdrawn from CDCS carriers (n=18) and normal parental subjects (n=18), all aged 0-34 years, aiming to set up a representative CDCS profile constructed from 33 targeted amino acids and biogenic amines. Methionine sulfoxide (MetO), nitrotyrosine and His were of particular concern with respect to CDCS redox balance. Increased serotonin (3-fold), methionine sulfoxide (2-fold), and Asp (50%) levels, and a little lower Orn (26%), citrulline (23%), Leu (29%), Val (20%), Ile (16%), Asn (17%), Gln (10%), Trp (15%), Thr (13%), His (12%), Phe (11%), Met (6%), and creatinine (35%) levels were found in the plasma of CDCS patients. Nitrotyrosine did not differ in normal and CDCS individuals, whereas the ratio Trp/Σtotal metabolites was 37% higher in the latter group. The accumulated metabolites may reflect disturbances in the redox balance, deficient purine biosynthesis, and altered behavior, whereas the amino acid abatement in the latter group may affect the homeostasis of the urea cycle, citric acid cycle, branched chain amino acid synthesis, Tyr and Trp metabolism and amino acid biosynthesis. The identification of enzymatic deficiencies leading to the amino acid burden in CDCS is further required for elucidating its molecular bases and eventually propose specific or mixed amino acid supplementation to newborn patients aiming to balance their metabolism. Support: FAPESP, CNPq, CAPES.

doi: 10.1016/j.freeradbiomed.2016.10.476

SfRBM / SFRRI 2016

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