@article{satheesh babu_petersen_iglesias-carres_paz_wankhade_neilson_anandh babu_2023, title={Blueberry intervention mitigates detrimental microbial metabolite trimethylamine N-oxide by modulating gut microbes}, ISSN={["1872-8081"]}, DOI={10.1002/biof.2014}, abstractNote={Gut microbes play a pivotal role in host physiology by producing beneficial or detrimental metabolites. Gut bacteria metabolize dietary choline and L-carnitine to trimethylamine (TMA) which is then converted to trimethylamine-N-oxide (TMAO). An elevated circulating TMAO is associated with diabetes, obesity, cardiovascular disease, and cancer in humans. In the present study, we investigated the effect of dietary blueberries and strawberries at a nutritional dosage on TMA/TMAO production and the possible role of gut microbes. Blueberry cohort mice received a control (C) or freeze-dried blueberry supplemented (CB) diet for 12 weeks and subgroups received an antibiotics cocktail (CA and CBA). Strawberry cohort mice received a control (N) or strawberry-supplemented (NS) diet and subgroups received antibiotics (NA and NSA). Metabolic parameters, choline, TMA, and TMAO were assessed in addition to microbial profiling and characterization of berry powders. Blueberry supplementation (equivalent to 1.5 human servings) reduced circulating TMAO in CB versus C mice (~48%) without changing choline or TMA. This effect was not mediated through alterations in metabolic parameters. Dietary strawberries did not reduce choline, TMA, or TMAO. Depleting gut microbes with antibiotics in these cohorts drastically reduced TMA and TMAO to not-quantified levels. Further, dietary blueberries increased the abundance of bacterial taxa that are negatively associated with circulating TMA/TMAO suggesting the role of gut microbes. Our phenolic profiling indicates that this effect could be due to chlorogenic acid and increased phenolic contents in blueberries. Our study provides evidence for considering dietary blueberries to reduce TMAO and prevent TMAO-induced complications.}, journal={BIOFACTORS}, author={Satheesh Babu, Adhini Kuppuswamy and Petersen, Chrissa and Iglesias-Carres, Lisard and Paz, Henry A. and Wankhade, Umesh D. and Neilson, Andrew P. and Anandh Babu, Pon Velayutham}, year={2023}, month={Nov} }
 @article{iglesias-carres_chadwick-corbin_sweet_neilson_2023, title={Dietary phenolics and their microbial metabolites are poor inhibitors of trimethylamine oxidation to trimethylamine N-oxide by hepatic flavin monooxygenase 3}, volume={120}, ISSN={["1873-4847"]}, DOI={10.1016/j.jnutbio.2023.109428}, abstractNote={High circulating levels of trimethylamine N-oxide (TMAO) have been associated with cardiovascular disease risk. TMAO is formed through a microbiome-host pathway utilizing primarily dietary choline as a substrate. Specific gut microbiota transform choline into trimethylamine (TMA), and, when absorbed, host hepatic flavin-containing monooxygenase 3 (FMO3) oxidizes TMA into TMAO. Chlorogenic acid and its metabolites reduce microbial TMA production in vitro. However, little is known regarding the potential for chlorogenic acid and its bioavailable metabolites to inhibit the last step: hepatic conversion of TMA to TMAO. We developed a screening methodology to study FMO3-catalyzed production of TMAO from TMA. HepG2 cells were unable to oxidize TMA into TMAO due to their lack of FMO3 expression. Although Hepa-1 cells did express FMO3 when pretreated with TMA and NADPH, they lacked enzymatic activity to produce TMAO. Rat hepatic microsomes contained active FMO3. Optimal reaction conditions were: 50 µM TMA, 0.2 mM NADPH, and 33 µL microsomes/mL reaction. Methimazole (a known FMO3 competitive substrate) at 200 µM effectively reduced FMO3-catalyzed conversion of TMA to TMAO. However, bioavailable chlorogenic acid metabolites did not generally inhibit FMO3 at physiological (1 µM) nor supra-physiological (50 µM) doses. Thus, the effects of chlorogenic acid in regulating TMAO levels in vivo are unlikely to occur through direct FMO3 enzyme inhibition. Potential effects on FMO3 expression remain unknown. Intestinal inhibition of TMA production and/or absorption are thus likely their primary mechanisms of action.}, journal={JOURNAL OF NUTRITIONAL BIOCHEMISTRY}, author={Iglesias-Carres, Lisard and Chadwick-Corbin, Sydney A. and Sweet, Michael G. and Neilson, Andrew P.}, year={2023}, month={Oct} }
 @article{babu_petersen_paz_iglesias-carres_li_zhong_neilson_wankhade_babu_2023, title={Gut Microbiota Depletion Using Antibiotics to Investigate Diet-Derived Microbial Metabolites: An Efficient Strategy}, ISSN={["1613-4133"]}, DOI={10.1002/mnfr.202300386}, abstractNote={SCOPE
Gut microbiota depletion using antibiotics in drinking water is a valuable tool to investigate the role of gut microbes and microbial metabolites in health and disease. However, there are challenges associated with this model. Animals avoid drinking water because of the antibiotic bitterness, which affects their metabolic health. The present study develops an efficient strategy to deplete gut microbes without affecting metabolic parameters.


METHODS AND RESULTS
Male C57BL/6J mice (7 weeks old) are fed a control (C) or high-fat (HF) diet. Subgroups of C and HF mice receive an antibiotic cocktail in drinking water (CA and HA). The antibiotic dosage is gradually increased so that the animals adapt to the taste of antibiotics. Metabolic parameters, gut microbiome, and microbial metabolites are assessed after 12 weeks treatment. Culture methods and 16s rRNA amplification confirm the depletion of gut microbes in antibiotic groups (CA and HA). Further, antibiotic treatment does not alter metabolic parameters (body weight, body fat, lean body mass, blood glucose, and glucose/insulin tolerance), whereas it suppresses the production of diet-derived microbial metabolites (trimethylamine and trimethylamine-N-oxide).


CONCLUSION
This strategy effectively depletes gut microbes and suppresses the production of microbial metabolites in mice without affecting their metabolic health.}, journal={MOLECULAR NUTRITION & FOOD RESEARCH}, author={Babu, Adhini Kuppuswamy Satheesh and Petersen, Chrissa and Paz, Henry A. and Iglesias-Carres, Lisard and Li, Ying and Zhong, Ying and Neilson, Andrew P. and Wankhade, Umesh D. and Babu, Pon Velayutham Anandh}, year={2023}, month={Dec} }
 @article{iglesias-carres_bruno_'antuono_linsalata_cardinali_neilson_2023, title={In vitro evidences of the globe artichoke antioxidant, cardioprotective and neuroprotective effects}, volume={107}, ISSN={["2214-9414"]}, DOI={10.1016/j.jff.2023.105674}, abstractNote={The chemical composition and in vitro biological activities of two globe artichoke varieties, Romanesco Siciliano (RS) and Spinoso Sardo (SS), were studied for their antioxidant activity and potential effects against age-related diseases. The globe artichoke extracts were subjected to three antioxidant activity tests, DPPH, ABTS and FRAP, and to the acetylcholinesterase neuro-related assay. The potential artichoke cardioprotective capacity was investigated by evaluating the inhibition of choline-d9 conversion into proatherogenic trimethylamine-d9 (TMA-d9), in an ex vivo-in vitro fecal fermentation model. In both globe artichoke varieties, the polyphenols profile was similar with chlorogenic and di-caffeoylquinic acids as the main polyphenols identified, although the amount changed with the variety (RS: 108.5; SS: 64.8 mg polyphenols/g DW). The RS sample gave the highest values of antioxidant and anti-acetylcholinesterase activities and mostly inhibited choline-d9 utilization (+28.0 % choline-d9 Area Under the Curve (AUC) compared to control), and TMA-d9 production (-31.9 % TMA-d9 AUC compared to control). All the reported results demonstrated that globe artichoke may provide potential cardioprotective, neuroprotective and antioxidant effects.}, journal={JOURNAL OF FUNCTIONAL FOODS}, author={Iglesias-Carres, Lisard and Bruno, Angelica and 'Antuono, Isabella and Linsalata, Vito and Cardinali, Angela and Neilson, Andrew P.}, year={2023}, month={Aug} }
 @article{iglesias-carres_racine_chadwick_nunn_kalambur_neilson_ferruzzi_2023, title={Mechanism of off-color formation in potato chips fried in oil systems containing ascorbic acid as a stabilizer}, volume={179}, ISSN={["1096-1127"]}, DOI={10.1016/j.lwt.2023.114682}, abstractNote={The use of alternative, green antioxidant (AOX) systems is demanded by consumers. Natural AOX systems pose significant challenges. For example, in frying applications, these AOX can negatively alter potato chip color, one of the most important traits in consumer selection. We evaluated the role of natural AOX systems containing ascorbic acid, tocopherols, and other antioxidants in amino acid-related undesirable color formation in fried potato chips. Results indicated that both oil phase AOX and potato factors are critical to generation of off-color formation in fried potato chips through Maillard type reactions. Ascorbic acid solubilization in oil and migration to the chip surface play key roles in observed off-color formation. However, multiple complex reactions may be responsible for color development, which may involve food matrix components. Contributions of AOX other than ascorbic acid appear minimal. Nevertheless, some browning can occur regardless of the presence of ascorbic acid. Color formation through glutamine occurred in the absence of ascorbic acid, but its presence greatly exacerbates color generation, while color generation via asparagine is barely modulated by ascorbic acid. AOX and free amino acid concentrations, temperature, and moisture are critical factors for controlling undesirable color formation during frying with natural oil AOX systems.}, journal={LWT-FOOD SCIENCE AND TECHNOLOGY}, author={Iglesias-Carres, Lisard and Racine, Kathryn C. and Chadwick, Sydney and Nunn, Candace and Kalambur, Sathya B. and Neilson, Andrew P. and Ferruzzi, Mario G.}, year={2023}, month={Apr} }
 @article{racine_iglesias-carres_herring_ferruzzi_kay_tessem_neilson_2022, title={Cocoa extract exerts sex-specific anti-diabetic effects in an aggressive type-2 diabetes model: A pilot study}, volume={626}, ISSN={["1090-2104"]}, DOI={10.1016/j.bbrc.2022.08.018}, abstractNote={Type 2 diabetes (T2D) is characterized by hyperglycemia and insulin resistance. Cocoa may slow T2D development and progression. This study employed male and female BTBR.Cg-Lepob/ob/WiscJ (ob/ob) and wild type (WT) controls to assess the potential for cocoa to ameliorate progressive T2D and compare responses between sexes. Mice received diet without (WT, ob/ob) or with cocoa extract (ob/ob + c) for 10 weeks. Acute cocoa reduced fasting hyperglycemia in females, but not males, after 2 weeks. Chronic cocoa supplementation (6-10 weeks) ameliorated hyperinsulinemia in males and worsened hyperlipidemia and hyperinsulinemia in females, yet also preserved and enhanced beta cell survival in females. The underlying mechanisms of these differences warrant further study. If sex differences are apparent in subsequent preclinical studies, clinical studies will be warranted to establish whether these differences are relevant in humans. Sex differences may need to be considered when designing human dietary interventions for T2D.}, journal={BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS}, author={Racine, Kathryn C. and Iglesias-Carres, Lisard and Herring, Jacob A. and Ferruzzi, Mario G. and Kay, Colin D. and Tessem, Jeffery S. and Neilson, Andrew P.}, year={2022}, month={Oct}, pages={205–210} }
 @article{iglesias-carres_racine_neilson_2022, title={Phenolic-rich beverages reduce bacterial TMA formation in an ex vivo-in vitro colonic fermentation model}, ISSN={["2042-650X"]}, DOI={10.1039/d2fo01159j}, abstractNote={The production of pro-atherogenic trimethylamine N-oxide (TMAO) is dependent on the gut microbiota metabolism of quaternary amines (i.e., choline) into trimethylamine (TMA). Nutritional strategies that target microbial conversion of choline into TMA could reduce cardiovascular disease and atherosclerosis burden by reducing subsequent formation of TMAO. This study aimed to evaluate (1) whether beverages rich in known inhibitors of TMA production (chlorogenic acid, catechin and epicatechin) can reduce TMA formation and (2) the effect of upper gastrointestinal digestion on efficacy. To do this, either raw or digested coffee, tea and cocoa beverages were evaluated for their TMA-d9 production inhibition in our ex vivo-in vitro fermentation model with human fecal slurries and choline-d9 substrate. Results showed that digestion was required to unlock the TMA-d9 production inhibition potential of coffee and cocoa beverages, and that teas did not possess a strong inhibition potential either digested or undigested. By fractionating digested bioactive beverages, we determined that those fractions rich in chlorogenic acid were the most bioactive. Overall, this study suggests that regular cocoa and coffee consumption could be a nutritional strategy able to reduce TMAO levels. In vivo studies should be carried out to confirm the potential of these beverages as strategies to inhibit TMA production.}, journal={FOOD & FUNCTION}, author={Iglesias-Carres, Lisard and Racine, Kathryn C. and Neilson, Andrew P.}, year={2022}, month={Jul} }
 @article{iglesias-carres_krueger_herring_tessem_neilson_2022, title={Potential of Phenolic Compounds and Their Gut Microbiota-Derived Metabolites to Reduce TMA Formation: Application of an In Vitro Fermentation High-Throughput Screening Model}, volume={70}, ISSN={["1520-5118"]}, DOI={10.1021/acs.jafc.2c00247}, abstractNote={Trimethylamine N-oxide (TMAO) is a pro-atherosclerotic product of dietary choline metabolism generated by a microbiome-host axis. The first step in this pathway is the enzymatic metabolism of choline to trimethylamine (TMA) by the gut microbiota. This reaction could be targeted to reduce atherosclerosis risk. We aimed to evaluate potential inhibitory effects of select dietary phenolics and their relevant gut microbial metabolites on TMA production via a human ex vivo-in vitro fermentation model. Various phenolics inhibited choline use and TMA production. The most bioactive compounds tested (caffeic acid, catechin, and epicatechin) reduced TMA-d9 formation (compared to control) by 57.5 ± 1.3 to 72.5 ± 0.4% at 8 h and preserved remaining choline-d9 concentrations by 194.1 ± 6.4 to 256.1 ± 6.3% at 8 h. These inhibitory effects were achieved without altering cell respiration or cell growth. However, inhibitory effects decreased at late fermentation times, which suggested that these compounds delay choline metabolism rather than completely inhibiting TMA formation. Overall, caffeic acid, catechin, and epicatechin were the most effective noncytotoxic inhibitors of choline use and TMA production. Thus, these compounds are proposed as lead bioactives to test in vivo.}, number={10}, journal={JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY}, author={Iglesias-Carres, Lisard and Krueger, Emily S. and Herring, Jacob A. and Tessem, Jeffery S. and Neilson, Andrew P.}, year={2022}, month={Mar}, pages={3207–3218} }
 @article{griffin_essenmacher_racine_iglesias-carres_tessem_smith_neilson_2021, title={Diet-induced obesity in genetically diverse collaborative cross mouse founder strains reveals diverse phenotype response and amelioration by quercetin treatment in 129S1/SvImJ, PWK/EiJ, CAST/PhJ, and WSB/EiJ mice}, volume={87}, ISSN={["1873-4847"]}, DOI={10.1016/j.jnutbio.2020.108521}, abstractNote={Significant evidence suggests protective effects of flavonoids against obesity in animal models, but these often do not translate to humans. One explanation for this disconnect is use of a few mouse strains (notably C57BL/6 J) in obesity studies. Obesity is a multifactorial disease. The underlying causes are not fully replicated by the high-fat C57BL/6 J model, despite phenotypic similarities. Furthermore, the impact of genetic factors on the activities of flavonoids is unknown. This study was designed to explore how diverse mouse strains respond to diet-induced obesity when fed a representative flavonoid. A subset of Collaborative Cross founder strains (males and females) were placed on dietary treatments (low-fat, high-fat, high-fat with quercetin, high-fat with quercetin and antibiotics) longitudinally. Diverse responses were observed across strains and sexes. Quercetin appeared to moderately blunt weight gain in male C57 and both sexes of 129S1/SvImJ mice, and slightly increased weight gain in female C57 mice. Surprisingly, quercetin dramatically blunted weight gain in male, but not female, PWK/PhJ mice. For female mice, quercetin blunted weight gain (relative to the high-fat phase) in CAST/PhJ, PWK/EiJ and WSB/EiJ mice compared to C57. Antibiotics did not generally result in loss of protective effects of quercetin. This highlights complex interactions between genetic factors, sex, obesity stimuli, and flavonoid intake, and the need to move away from single inbred mouse models to enhance translatability to diverse humans. These data justify use of genetically diverse Collaborative Cross and Diversity Outbred models which are emerging as invaluable tools in the field of personalized nutrition.}, journal={JOURNAL OF NUTRITIONAL BIOCHEMISTRY}, author={Griffin, Laura E. and Essenmacher, Lauren and Racine, Kathryn C. and Iglesias-Carres, Lisard and Tessem, Jeffery S. and Smith, Susan M. and Neilson, Andrew P.}, year={2021}, month={Jan} }
 @article{iglesias-carres_hughes_steele_ponder_davy_neilson_2021, title={Use of dietary phytochemicals for inhibition of trimethylamine N-oxide formation}, volume={91}, ISSN={["1873-4847"]}, DOI={10.1016/j.jnutbio.2021.108600}, abstractNote={Trimethylamine-N-oxide (TMAO) has been reported as a risk factor for atherosclerosis development, as well as for other cardiovascular disease (CVD) pathologies. The objective of this review is to provide a useful summary on the use of phytochemicals as TMAO-reducing agents. This review discusses the main mechanisms by which TMAO promotes CVD, including the modulation of lipid and bile acid metabolism, and the promotion of endothelial dysfunction and oxidative stress. Current knowledge on the available strategies to reduce TMAO formation are discussed, highlighting the effect and potential of phytochemicals. Overall, phytochemicals (i.e., phenolic compounds or glucosinolates) reduce TMAO formation by modulating gut microbiota composition and/or function, inhibiting host's capacity to metabolize TMA to TMAO, or a combination of both. Perspectives for design of future studies involving phytochemicals as TMAO-reducing agents are discussed. Overall, the information provided by this review outlines the current state of the art of the role of phytochemicals as TMAO reducing agents, providing valuable insight to further advance in this field of study.}, journal={JOURNAL OF NUTRITIONAL BIOCHEMISTRY}, author={Iglesias-Carres, Lisard and Hughes, Michael D. and Steele, Cortney N. and Ponder, Monica A. and Davy, Kevin P. and Neilson, Andrew P.}, year={2021}, month={May} }
 @article{iglesias-carres_neilson_2021, title={Utilizing preclinical models of genetic diversity to improve translation of phytochemical activities from rodents to humans and inform personalized nutrition}, ISSN={["2042-650X"]}, DOI={10.1039/d1fo02782d}, abstractNote={Mouse models are an essential tool in different areas of research, including nutrition and phytochemical research. Traditional inbred mouse models have allowed the discovery of therapeutical targets and mechanisms of action and expanded our knowledge of health and disease. However, these models lack the genetic variability typically found in human populations, which hinders the translatability of the results found in mice to humans. The development of genetically diverse mouse models, such as the collaborative cross (CC) or the diversity outbred (DO) models, has been a useful tool to overcome this obstacle in many fields, such as cancer, immunology and toxicology. However, these tools have not yet been widely adopted in the field of phytochemical research. As demonstrated in other disciplines, use of CC and DO models has the potential to provide invaluable insights for translation of phytochemicals from rodents to humans, which are desperately needed given the challenges and numerous failed clinical trials in this field. These models may prove informative for personalized use of phytochemicals in humans, including: predicting interindividual variability in phytochemical bioavailability and efficacy, identifying genetic loci or genes governing response to phytochemicals, identifying phytochemical mechanisms of action and therapeutic targets, and understanding the impact of genetic variability on individual response to phytochemicals. Such insights would prove invaluable for personalized implementation of phytochemicals in humans. This review will focus on the current work performed with genetically diverse mouse populations, and the research opportunities and advantages that these models can offer to phytochemical research.}, journal={FOOD & FUNCTION}, author={Iglesias-Carres, Lisard and Neilson, Andrew P.}, year={2021}, month={Oct} }