TY - JOUR TI - Techno-economic analysis (TEA) of low-moisture anhydrous ammonia (LMAA) pretreatment method for corn stover AU - Yang, Minliang AU - Rosentrater, Kurt A. T2 - Industrial Crops and Products AB - Techno-economic analysis (TEA) plays an important role in assessing economic performance and potential market acceptance for new technologies. Previous work has shown that the construction and operation of a cellulosic bioethanol plant can be very expensive. One of the largest cost categories is pretreatment processing. The purpose of this study was to conduct a detailed cost analysis to assess low moisture anhydrous ammonia (LMAA) pretreatment process at the commercial-scale, and to estimate the breakeven point in large-scale production. In this study, capital expenses, including annualized purchase and installation fees, and annual operating costs associated with each unit operation were determined. The lowest unit cost obtained from this study was $3.86/gal, but it was still high compared with current gasoline price. DA - 2015/12// PY - 2015/12// DO - 10.1016/j.indcrop.2015.06.023 VL - 76 SP - 55-61 J2 - Industrial Crops and Products LA - en OP - SN - 0926-6690 UR - http://dx.doi.org/10.1016/j.indcrop.2015.06.023 DB - Crossref ER - TY - JOUR TI - Corrigendum to “Enzymatic synthesis of substituted epicatechins for bioactivity studies in neurological disorders” [Biochem. Biophys. Res. Commun. 417 (1) (6 January 2012) 457–461] AU - Blount, Jack W. AU - Ferruzzi, Mario AU - Raftery, Dan AU - Pasinetti, Giulio M. AU - Dixon, Richard A. T2 - Biochemical and Biophysical Research Communications DA - 2015/5// PY - 2015/5// DO - 10.1016/J.BBRC.2015.02.142 VL - 460 IS - 2 SP - 489 J2 - Biochemical and Biophysical Research Communications LA - en OP - SN - 0006-291X UR - http://dx.doi.org/10.1016/J.BBRC.2015.02.142 DB - Crossref ER - TY - CONF TI - Sulfurophane represses matrix-degrading proteases and protects cartilage from destruction in vitro and in vivo AU - Davidson, R. AU - Jupp, O. AU - de Ferrars, R. AU - Kay, C. AU - Culley, K. AU - Norton, R. AU - Driscoll, C. AU - Vincent, T. AU - Donell, S. AU - Bao, Y. T2 - Joint meeting of British Societies for Matrix Biology and Developmental Biology C2 - 2015/// C3 - International Journal of Experimental Pathology CY - University of East Anglia (Norwich) DA - 2015/// PY - 2014/9/1/ VL - 96 SP - A24 M1 - 2 ER - TY - JOUR TI - Carbohydrate and Phytochemical Digestibility in Pasta AU - Camelo-Méndez, Gustavo A. AU - Ferruzzi, Mario G. AU - González-Aguilar, Gustavo A. AU - Bello-Pérez, Luis A. T2 - Food Engineering Reviews DA - 2015/3/10/ PY - 2015/3/10/ DO - 10.1007/S12393-015-9117-Z VL - 8 IS - 1 SP - 76-89 J2 - Food Eng Rev LA - en OP - SN - 1866-7910 1866-7929 UR - http://dx.doi.org/10.1007/S12393-015-9117-Z DB - Crossref KW - Pasta products KW - Raw materials KW - Processing technologies KW - Carbohydrates digestibility KW - Phytochemicals KW - Glycemic index ER - TY - JOUR TI - The bioactivity of flavonoids is likely the result of cumulative low exposure to a variety of structurally similar phenolic metabolites AU - Kay, Colin AU - de Gesso, Jessica AU - Warner, Emily AU - Amin, Hiren AU - de Ferrars, Rachel AU - Edwards, Michael AU - Zang, Qing AU - O'Hagan, David AU - O'Connell, Maria T2 - The FASEB Journal AB - Activities of 21 flavonoid metabolites were investigated relative to their unmetabolized structures, utilizing 20 assays of vascular reactivity, inflammation and cellular adhesion, in endothelial (HUVEC), smooth muscle (AVSMC) and monocyte (THP‐1) cells, at physiologically relevant concentrations (0.1‐10µM). Unmetabolized flavonoids were generally less active than their phenolic metabolites. In vascular screens no metabolites altered expression of endothelin‐1 or nitric oxide, 3 metabolites induced eNOS, 2metabolites reduced angiotensin‐II induced superoxide, no metabolites induced NOX or p47phox, 1 metabolite upregulated p22phox and 3 metabolites induced HO‐1. In vascular inflammation and adhesion screens in HUVEC, 10, 9 and 6 metabolites reduced IL‐6 following CD40L, oxLDL or TNF‐α stimulation, respectively. 10 and 6 metabolites reduced sVCAM‐1 following CD40L or TNF‐α stimulation, respectively. 2 metabolites attenuated IL‐1β‐stimulated phosphorylation of NF‐κB p65, 1 metabolite induced Nrf2. In monocyte activity assays, 5 metabolites reduced LPS‐induced TNF‐α, 1 metabolite induced IL1B, no metabolites were active on HO‐1, tissue factor, NFkB or nfKB. The evidence suggests that bioactivity of flavonoids in vivo results from low exposure to a variety of structurally similar metabolites modifying inflammation and cellular adhesion pathways. Support: UK Biotechnology and Biological Sciences Research Council (BB/I006028/1, BB/H004963/1). DA - 2015/// PY - 2015/// DO - 10.1096/fasebj.29.1_supplement.118.4 VL - 29 IS - Supplement 1 SP - 118.4 UR - https://doi.org/10.1096/fasebj.29.1_supplement.118.4 ER - TY - JOUR TI - Method to harness bioactive secondary metabolites from intact Quinoa seeds with implications for chronic disease prevention AU - Graf, BL AU - Poulev, A AU - Kuhn, P AU - Esposito, D AU - Lila, MA AU - Raskin, I T2 - Planta Medica AB - Quinoa (Chenopodium quinoa Willd.) is an Andean seed crop rich in bioactive phytochemicals, including phytoecdysteroids (PE) and flavonoid glycosides (FG). Innovations designed to harness the pharmacological value of quinoa through simple, food-grade technologies may facilitate the development of functional foods and nutraceuticals to combat global human health challenges. We optimized a method to leach and concentrate quinoa bioactives from intact (un-macerated) quinoa seeds into aqueous ethanol, yielding a complex phytochemical mixture termed quinoa leachate (QL). QL, comprised of 1.0% PE and 2.6% FG, contained essentially all PE and FG available in the initial seeds compared with traditional extraction of macerated seed powder (567.6 µg PE and 540.9 µg FG/g seed). QL significantly lowered fasting blood glucose in obese, hyperglycemic C57Bl/6J mice and significantly attenuated lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) production in human dermal fibroblasts. Quinoa seed leaching provides an efficient means to produce a food-grade mixture that may be applicable for the treatment and prevention of chronic, complex diseases, while rendering intact post-leached seeds available for additional uses in food. DA - 2015/6/25/ PY - 2015/6/25/ DO - 10.1055/S-0035-1556449 VL - 81 IS - 11 J2 - Planta Med LA - en OP - SN - 0032-0943 1439-0221 UR - http://dx.doi.org/10.1055/S-0035-1556449 DB - Crossref ER - TY - JOUR TI - New functionally-enhanced soy proteins as food ingredients with anti-viral activity AU - Turmagambetova, Aizhan Sabirzhanovna AU - Sokolova, Nadezhda Sergeevna AU - Bogoyavlenskiy, Andrey Pavlinovich AU - Berezin, Vladimir Eleazarovich AU - Lila, Mary Ann AU - Cheng, Diana M. AU - Dushenkov, Vyacheslav T2 - VirusDisease AB - Respiratory viruses are a major public health problem because of their prevalence and high morbidity rate leading to considerable social and economic implications. Cranberry has therapeutic potential attributed to a comprehensive list of phytochemicals including anthocyanins, flavonols, and unique A-type proanthocyanidins. Soy flavonoids, including isoflavones, have demonstrated anti-viral effects in vitro and in vivo. Recently, it was demonstrated that edible proteins can efficiently sorb and concentrate cranberry polyphenols, including anthocyanins and proanthocyanins, providing greatly stabilized matrices suitable for food products. The combination of cranberry and soy phytoactives may be an effective dietary anti-viral resource. Anti-viral properties of both cranberry juice-enriched and cranberry pomace polyphenol-enriched soy protein isolate (CB-SPI and CBP-SPI) were tested against influenza viruses (H7N1, H5N3, H3N2), Newcastle disease virus and Sendai virus in vitro and in ovo. In our experiments, preincubation with CB-SPI or CBP-SPI resulted in inhibition of virus adsorption to chicken red blood cells and reduction in virus nucleic acid content up to 16-fold, however, CB-SPI and CBP-SPI did not affect hemagglutination. Additionally, CB-SPI and CBP-SPI inhibited viral replication and infectivity more effectively than the commercially available anti-viral drug Amizon. Results suggest CB-SPI and CBP-SPI may have preventative and therapeutic potential against viral infections that cause diseases of the respiratory and gastro-intestinal tract. DA - 2015/8/15/ PY - 2015/8/15/ DO - 10.1007/S13337-015-0268-6 VL - 26 IS - 3 SP - 123-132 J2 - VirusDis. LA - en OP - SN - 2347-3584 2347-3517 UR - http://dx.doi.org/10.1007/S13337-015-0268-6 DB - Crossref ER - TY - JOUR TI - Novel application of brain-targeting polyphenol compounds in sleep deprivation-induced cognitive dysfunction AU - Zhao, Wei AU - Wang, Jun AU - Bi, Weina AU - Ferruzzi, Mario AU - Yemul, Shrishailam AU - Freire, Daniel AU - Mazzola, Paolo AU - Ho, Lap AU - Dubner, Lauren AU - Pasinetti, Giulio Maria T2 - Neurochemistry International AB - Sleep deprivation produces deficits in hippocampal synaptic plasticity and hippocampal-dependent memory storage. Recent evidence suggests that sleep deprivation disrupts memory consolidation through multiple mechanisms, including the down-regulation of the cAMP-response element-binding protein (CREB) and of mammalian target of rapamycin (mTOR) signaling. In this study, we tested the effects of a Bioactive Dietary Polyphenol Preparation (BDPP), comprised of grape seed polyphenol extract, Concord grape juice, and resveratrol, on the attenuation of sleep deprivation-induced cognitive impairment. We found that BDPP significantly improves sleep deprivation-induced contextual memory deficits, possibly through the activation of CREB and mTOR signaling pathways. We also identified brain-available polyphenol metabolites from BDPP, among which quercetin-3-O-glucuronide activates CREB signaling and malvidin-3-O-glucoside activates mTOR signaling. In combination, quercetin and malvidin-glucoside significantly attenuated sleep deprivation-induced cognitive impairment in -a mouse model of acute sleep deprivation. Our data suggests the feasibility of using select brain-targeting polyphenol compounds derived from BDPP as potential therapeutic agents in promoting resilience against sleep deprivation-induced cognitive dysfunction. DA - 2015/10// PY - 2015/10// DO - 10.1016/J.NEUINT.2015.07.023 VL - 89 SP - 191-197 J2 - Neurochemistry International LA - en OP - SN - 0197-0186 UR - http://dx.doi.org/10.1016/J.NEUINT.2015.07.023 DB - Crossref KW - Sleep deprivation KW - Polyphenols KW - Cognitive dysfunction KW - Resilience KW - Memory consolidation ER - TY - JOUR TI - Role of intestinal microbiota in the generation of polyphenol-derived phenolic acid mediated attenuation of Alzheimer's disease β-amyloid oligomerization AU - Wang, Dongjie AU - Ho, Lap AU - Faith, Jeremiah AU - Ono, Kenjiro AU - Janle, Elsa M. AU - Lachcik, Pamela J. AU - Cooper, Bruce R. AU - Jannasch, Amber H. AU - D'Arcy, Bruce R. AU - Williams, Barbara A. AU - Ferruzzi, Mario G. AU - Levine, Samara AU - Zhao, Wei AU - Dubner, Lauren AU - Pasinetti, Giulio M. T2 - Molecular Nutrition & Food Research AB - Scope Grape seed polyphenol extract (GSPE) is receiving increasing attention for its potential preventative and therapeutic roles in Alzheimer's disease (AD) and other age‐related neurodegenerative disorders. The intestinal microbiota is known to actively convert many dietary polyphenols, including GSPE, to phenolic acids. There is limited information on the bioavailability and bioactivity of GSPE‐derived phenolic acid in the brain. Methods and results We orally administered GSPE to rats and investigated the bioavailability of 12 phenolic acids known to be generated by microbiota metabolism of anthocyanidins. GSPE treatment significantly increased the content of two of the phenolic acids in the brain: 3‐hydroxybenzoic acid and 3‐(3´‐hydroxyphenyl)propionic acid, resulting in the brain accumulations of the two phenolic acids at micromolar concentrations. We also provided evidence that 3‐hydroxybenzoic acid and 3‐(3´‐hydroxyphenyl)propionic acid potently interfere with the assembly of β‐amyloid peptides into neurotoxic β‐amyloid aggregates that play key roles in AD pathogenesis. Conclusion Our observation suggests important contribution of the intestinal microbiota to the protective activities of GSPE (as well as other polyphenol preparations) in AD. Outcomes from our studies support future preclinical and clinical investigations exploring the potential contributions of the intestinal microbiota in protecting against the onset/progression of AD and other neurodegenerative conditions. DA - 2015/4/27/ PY - 2015/4/27/ DO - 10.1002/MNFR.201400544 VL - 59 IS - 6 SP - 1025-1040 J2 - Mol. Nutr. Food Res. LA - en OP - SN - 1613-4125 UR - http://dx.doi.org/10.1002/MNFR.201400544 DB - Crossref KW - -Amyloid KW - Bioavailability KW - Grape seed polyphenol extract KW - Phenolic metabolites KW - Proanthocyanidins ER - TY - JOUR TI - Impact of a Service-Learning-Based Community Nutrition Course on Students’ Nutrition Teaching Self-Efficacy AU - Cooke, Natalie K. AU - Ash, S.L. AU - Nietfeld, J. AU - Fogleman, A. AU - Goodell, L.S. T2 - Journal of Nutrition Education and Behavior AB - The purpose of this study was to determine the effect of a service-learning (SL) course on student self-efficacy (SE) in teaching nutrition in the community. Students in the experimental group (n=20) were enrolled in a Community Nutrition SL course, and students in the control group were in a Public Health Nutrition non-SL course (n=63). To build SE, students need to practice their skills in an appropriate setting. Service-learning, a pedagogy combining academic learning with service in the community, is an ideal framework for this preparation. The SL course consisted of a 6-week pre-SL training designed to increase SE through skills-based training, observation, and practice and a 6-week SL experience in which students taught a pre-packaged curriculum in the community. Researchers developed the Self-Efficacy in Teaching Nutrition in the Community (SET-NC) survey to evaluate changes in student SE for teaching nutrition in the community. Experimental and control groups took the SET-NC survey three times during the semester. Using time 1 as a covariant, results from a repeated measures ANCOVA indicated the experimental group had significantly higher SE scores than the control group at both time 2 (p=0.008) and time 3 (p<0.001). The SL course increased students’ SE in teaching nutrition in the community, and other nutrition and dietetics programs may benefit from a similar SL course design. Researchers are currently validating the SET-NC survey in a nationwide population to provide a useful tool for measuring students’ SE in teaching nutrition in the community. DA - 2015/7// PY - 2015/7// DO - 10.1016/J.JNEB.2015.04.013 VL - 47 IS - 4 SP - S4 J2 - Journal of Nutrition Education and Behavior LA - en OP - SN - 1499-4046 UR - http://dx.doi.org/10.1016/J.JNEB.2015.04.013 DB - Crossref ER - TY - JOUR TI - Physicochemical, bioactive and functional evaluation of the exotic fruits Opuntia ficus-indica AND Pilosocereus pachycladus Ritter from the Brazilian caatinga T2 - Journal of Food Science and Technology AB - The Brazilian caatinga is characterized as one of the most populated and biologically diverse semi-arid regions in the world. Two exotic fruits collected in this unique bioma, the prickly pear (Opuntia ficus-indica) and facheiro (Philosocereus pachycladus Ritter), were investigated in regard to their physicochemical, bioactive and functional characteristics. Four different extracts were prepared and investigated: water (W), ethanol/water 70:100 (E70), ethanol/water 80:100 (E80), and ethanol only (E100). The betalain compounds were investigated using the LC-DAD-ESI-MS technique. The prickly pear fruits are sweeter and less acidic when compared to facheiro fruits, but they also have lower total solids, ash and protein. The total phenolic content (TPC) of water and ethanolic extracts ranged from 8.89 to 12.34 mg GAE/100 g and 82.23 to 107.67 mg GAE/100 g for Opuntia and facheiro fruits, respectively. The identification of betanin and isobetanin derivatives, as well as the TPC, antioxidant and antienzymatic activities of facheiro fruits were reported for the first time. All extracts presented some degree of alpha-amylase and alpha-glucosidase inhibition, with the exception of the facheiro water extracts. DA - 2015/11// PY - 2015/11// DO - 10.1007/s13197-015-1821-4 UR - http://dx.doi.org/10.1007/s13197-015-1821-4 KW - Caatinga KW - Cactaceae fruits KW - Phenolics KW - Betalains ER - TY - JOUR TI - Synthesis and Quantitative Analysis of Plasma-Targeted Metabolites of Catechin and Epicatechin AU - Blount, Jack W. AU - Redan, Benjamin W. AU - Ferruzzi, Mario G. AU - Reuhs, Bradley L. AU - Cooper, Bruce R. AU - Harwood, John S. AU - Shulaev, Vladimir AU - Pasinetti, Giulio AU - Dixon, Richard A. T2 - Journal of Agricultural and Food Chemistry AB - Grape seed polyphenolic extract (GSPE) rich in the flavan-3-ols (+)-catechin and (-)-epicatechin beneficially modulates Alzheimer's Disease phenotypes in animal models. The parent molecules in the extract are converted to a series of methylated and glucuronidated derivatives. To fully characterize these metabolites and establish a robust quantitative assay of their levels in biological fluids, we have implemented a partial synthetic approach utilizing chemical methylation followed by enzymatic glucuronidation. Liquid chromatography/time-of-flight mass spectrometry (LC-TOF-MS) and nuclear magnetic resonance (NMR) spectroscopy were used to assign unequivocal structures to the compounds. An analytical method using solid-phase extraction and LC-MS/MS in selective reaction monitoring mode (SRM) was validated for their quantitation in plasma. These studies provide a basis for improvements in future work on the bioavailability, metabolism, and mechanism of action of metabolites derived from dietary flavan-3-ols in a range of interventions. DA - 2015/2/20/ PY - 2015/2/20/ DO - 10.1021/JF505922B VL - 63 IS - 8 SP - 2233-2240 J2 - J. Agric. Food Chem. LA - en OP - SN - 0021-8561 1520-5118 UR - http://dx.doi.org/10.1021/JF505922B DB - Crossref KW - catechin KW - glucuronosylation KW - O-methylation KW - phase II metabolites KW - semisynthesis KW - structure determination KW - two-site validation ER - TY - JOUR TI - Dietary Phenolic Compounds Selectively Inhibit the Individual Subunits of Maltase-Glucoamylase and Sucrase-Isomaltase with the Potential of Modulating Glucose Release AU - Simsek, Meric AU - Quezada-Calvillo, Roberto AU - Ferruzzi, Mario G. AU - Nichols, Buford L. AU - Hamaker, Bruce R. T2 - Journal of Agricultural and Food Chemistry AB - In this study, it was hypothesized that dietary phenolic compounds selectively inhibit the individual C- and N-terminal (Ct, Nt) subunits of the two small intestinal α-glucosidases, maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI), for a modulated glycemic carbohydrate digestion. The inhibition by chlorogenic acid, caffeic acid, gallic acid, (+)-catechin, and (-)-epigallocatechin gallate (EGCG) on individual recombinant human Nt-MGAM and Nt-SI and on mouse Ct-MGAM and Ct-SI was assayed using maltose as the substrate. Inhibition constants, inhibition mechanisms, and IC50 values for each combination of phenolic compound and enzymatic subunit were determined. EGCG and chlorogenic acid were found to be more potent inhibitors for selectively inhibiting the two subunits with highest activity, Ct-MGAM and Ct-SI. All compounds displayed noncompetitive type inhibition. Inhibition of fast-digesting Ct-MGAM and Ct-SI by EGCG and chlorogenic acid could lead to a slow, but complete, digestion of starch for improved glycemic response of starchy foods with potential health benefit. DA - 2015/4/13/ PY - 2015/4/13/ DO - 10.1021/JF505425D VL - 63 IS - 15 SP - 3873-3879 J2 - J. Agric. Food Chem. LA - en OP - SN - 0021-8561 1520-5118 UR - http://dx.doi.org/10.1021/JF505425D DB - Crossref KW - alpha-glucosidases KW - inhibition KW - maltase-glucoamylase KW - phenolics KW - sucrase-isomaltase ER - TY - JOUR TI - Microwave-assisted extraction of lycopene in tomato peels: Effect of extraction conditions on all-trans and cis-isomer yields AU - Ho, K.K.H.Y. AU - Ferruzzi, M.G. AU - Liceaga, A.M. AU - San Martín-González, M.F. T2 - LWT - Food Science and Technology AB - Lycopene is the primary carotenoid in tomato peels, a processing byproduct, and can be used as a natural color or bioactive ingredient. Unfortunately, extractions are inefficient as lycopene is extremely nonpolar and susceptible to degradation. As a rapid technique, microwave-assisted extraction (MAE) potentially offers efficient lycopene recovery. Thus, the objectives of this research were to: 1) optimize MAE of lycopene from tomato peels and 2) evaluate the effect of treatment on all-trans and isomer yields. Response surface methodology (RSM) was employed to optimize lycopene extraction with solvent ratio solid–liquid ratios, microwave power, and delivered energy equivalents as factors. High performance liquid chromatography with a diode array detector (HPLC-DAD) was used for isomer separation and quantification. Optimum MAE conditions were determined as: 0:10 solvent ratio at 400 W with a yield of 13.592 mg/100 g of extracted all-trans-lycopene. RSM suggested that ethyl acetate was a better MAE solvent for lycopene recovery as compared to hexane, which overall extracted less lycopene. HPLC-DAD indicated that MAE significantly improved all-trans and total lycopene yields, while conventional extraction demonstrated higher proportions of cis-isomer yields. Additionally, electron micrographs showed that significant structural disruption occurred in MAE-treated samples, possibly allowing for the improved lycopene extraction. DA - 2015/6// PY - 2015/6// DO - 10.1016/J.LWT.2014.12.061 VL - 62 IS - 1 SP - 160-168 J2 - LWT - Food Science and Technology LA - en OP - SN - 0023-6438 UR - http://dx.doi.org/10.1016/J.LWT.2014.12.061 DB - Crossref KW - All-trans-lycopene KW - Cis-isomers KW - Microwave-assisted extraction KW - Response surface methodology ER - TY - JOUR TI - Correction to Survey of Polyphenol Constituents in Grapes and Grape-Derived Products AU - Xu, Yanping AU - Simon, James E. AU - Welch, Cara AU - Wightman, JoLynne D. AU - Ferruzzi, Mario G. AU - Ho, Lap AU - Pasinetti, Giulio M. AU - Wu, Qingli T2 - Journal of Agricultural and Food Chemistry AB - ADVERTISEMENT RETURN TO ISSUEPREVAddition/CorrectionORIGINAL ARTICLEThis notice is a correctionCorrection to Survey of Polyphenol Constituents in Grapes and Grape-Derived ProductsYanping Xu, James E. Simon, Cara Welch, JoLynne D. Wightman, Mario G. Ferruzzi, Lap Ho, Giulio M. Pasinetti, and Qingli Wu*Cite this: J. Agric. Food Chem. 2015, 63, 14, 3804Publication Date (Web):April 6, 2015Publication History Published online6 April 2015Published inissue 15 April 2015https://doi.org/10.1021/acs.jafc.5b01552Copyright © 2015 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views468Altmetric-Citations1LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (110 KB) Get e-Alertsclose Get e-Alerts DA - 2015/4/6/ PY - 2015/4/6/ DO - 10.1021/ACS.JAFC.5B01552 VL - 63 IS - 14 SP - 3804-3804 J2 - J. Agric. Food Chem. LA - en OP - SN - 0021-8561 1520-5118 UR - http://dx.doi.org/10.1021/ACS.JAFC.5B01552 DB - Crossref ER - TY - JOUR TI - Influence of molecular weight on intracellular antioxidant activity of invasive silver carp ( Hypophthalmichthys molitrix ) protein hydrolysates AU - Malaypally, Sravanthi P. AU - Liceaga, Andrea M. AU - Kim, Kee-Hong AU - Ferruzzi, Mario AU - Martin, Fernanda San AU - Goforth, Reuben R. T2 - Journal of Functional Foods AB - Protein hydrolysates from underutilized silver carp (SPH) were prepared using Flavourzyme (F-15 to F-60) and Alcalase (A-15 to A-60) at 15, 30, 45 and 60 min, respectively. SPH F-30 and A-60 showed promising chemical-based antioxidant activity and were further fractionated according to size to evaluate caco-2 cell based antioxidant activity. F-30 and A-60 peptide fractions with <3 kDa (F-30 < 3, A-60 < 3) showed higher cell-based antioxidant activity under stressed and non-stressed conditions. Further, IC50 values of F-30 < 3 (1–3 mg/mL) was lower than A-60 < 3 fractions (4 to 12 mg/mL), indicating higher cellular antioxidant activity of F-30 < 3 compared to A-60 < 3 under all conditions. The presence of active peptides with desired amino acid sequence in F-30 < 3 compared to A-60 < 3 may have contributed to its higher cellular antioxidant activity. Overall, SPH exhibited antioxidant capacity, hence using an underutilized, invasive fish for environmental and economic gain in the form of promising functional ingredients. DA - 2015/10// PY - 2015/10// DO - 10.1016/J.JFF.2014.06.011 VL - 18 SP - 1158-1166 J2 - Journal of Functional Foods LA - en OP - SN - 1756-4646 UR - http://dx.doi.org/10.1016/J.JFF.2014.06.011 DB - Crossref KW - Protein hydrolysates KW - Intracellular antioxidant activity KW - Molecular weight KW - Peptide sequence ER - TY - JOUR TI - Thermal degradation of green tea flavan-3-ols and formation of hetero- and homocatechin dimers in model dairy beverages AU - Song, Brian J. AU - Manganais, Chris AU - Ferruzzi, Mario G. T2 - Food Chemistry AB - Interactions between polyphenols and macromolecules may impact polyphenol stability and bioavailability from foods. The impact of milk on tea flavan-3-ol stability to thermal treatment was investigated. Single strength (36.2 protein per L), quarter strength (9.0 g protein per L) milk, and control model beverages were incubated with epigallocatechin gallate and green tea extract at 62 or 37 °C for 180 min. Intact flavan-3-ols and select auto-oxidation products [theasinesins (THSNs) and P-2 dimers] were quantified by LC–MS. Generally, greater polyphenol to protein ratios increased first order degradation rates, consequently decreasing formation of oxidation products. The presence of galloyl and hydroxy moieties was associated with higher stability of monomeric flavan-3-ols with increasing protein concentrations suggesting potential for protein affinity to stabilise flavan-3-ols to thermal treatment. Absence of these moieties led to no observable improvements in stability. These results suggest that protein interactions may be useful in stabilising flavan-3-ols through thermal processing. DA - 2015/4// PY - 2015/4// DO - 10.1016/J.FOODCHEM.2014.10.026 VL - 173 SP - 305-312 J2 - Food Chemistry LA - en OP - SN - 0308-8146 UR - http://dx.doi.org/10.1016/J.FOODCHEM.2014.10.026 DB - Crossref KW - Flavan-3-ols KW - Thermal stability KW - Oxidation KW - Theasinesin KW - P-2 dimer KW - Protein-polyphenol interaction ER - TY - JOUR TI - Chemical investigation of commercial grape seed derived products to assess quality and detect adulteration AU - Villani, Tom S. AU - Reichert, William AU - Ferruzzi, Mario G. AU - Pasinetti, Giulio M. AU - Simon, James E. AU - Wu, Qingli T2 - Food Chemistry AB - Fundamental concerns in quality control arise due to increasing use of grape seed extract (GSE) and the complex chemical composition of GSE. Proanthocyanidin monomers and oligomers are the major bioactive compounds in GSE. Given no standardized criteria for quality, large variation exists in the composition of commercial GSE supplements. Using HPLC/UV/MS, 21 commercial GSE containing products were purchased and chemically profiled, major compounds quantitated, and compared against authenticated grape seed extract, peanut skin extract, and pine bark extract. The antioxidant capacity and total polyphenol content for each sample was also determined and compared using standard techniques. Nine products were adulterated, found to contain peanut skin extract. A wide degree of variability in chemical composition was detected in commercial products, demonstrating the need for development of quality control standards for GSE. A TLC method was developed to allow for rapid and inexpensive detection of adulteration in GSE by peanut skin. DA - 2015/3// PY - 2015/3// DO - 10.1016/J.FOODCHEM.2014.08.084 VL - 170 SP - 271-280 J2 - Food Chemistry LA - en OP - SN - 0308-8146 UR - http://dx.doi.org/10.1016/J.FOODCHEM.2014.08.084 DB - Crossref KW - Grape seed extract KW - LC-MS KW - Polyphenol KW - Proanthocyanidins KW - Adulteration KW - Quality control ER - TY - JOUR TI - Plasma bioavailability and regional brain distribution of polyphenols from apple/grape seed and bilberry extracts in a young swine model AU - Chen, Tzu-Ying AU - Kritchevsky, Janice AU - Hargett, Katherine AU - Feller, Kathryn AU - Klobusnik, Ryan AU - Song, Brian J. AU - Cooper, Bruce AU - Jouni, Zeina AU - Ferruzzi, Mario G. AU - Janle, Elsa M. T2 - Molecular Nutrition & Food Research AB - The pharmacokinetics, bioavailability, and regional brain distribution of polyphenols from apple-grape seed extract (AGSE) mixture and bilberry extract were studied after 3 weeks of dosing in weanling pigs.Weanling piglets were treated for 3 weeks with extracts of (AGSE) or bilberry extracts, using a physiological (27.5 mg/kg) or supplement (82.5 mg/kg) dose. A 24-h pharmacokinetic study was conducted and brain tissue was harvested. Major flavan-3-ol and flavonol metabolites including catechin-O-β-glucuronide, epicatechin-O-β-glucuronide, 3'O-methyl-catechin-O-β-glucuronide, 3'O-methyl-epicatechin-O-β-glucuronide, quercetin-O-β-glucuronide, and O-methyl-quercetin-O-β-glucuronide were analyzed in plasma, urine, and regional brain extracts from AGSE groups. Anthocyanidin-O-galactosides and O-glucosides of delphinidin (Del), cyanidin (Cyn), petunidin (Pet), peonidin (Peo), and malvidin (Mal) were analyzed in plasma, urine, and brain extracts from bilberry groups.Significant plasma dose-dependence was observed in flavan-3-ol metabolites of the AGSE group and in Mal, Del and Cyn galactosides and Pet, Peo, and Cyn glucosides of the bilberry groups. In the brain, a significant dose dependence was found in the cerebellum and frontal cortex in all major flavan-3-ol metabolites. All anthocyanidin glycosides, except for delphinidin, showed a dose-dependent increase in the cerebellum. DA - 2015/10/28/ PY - 2015/10/28/ DO - 10.1002/MNFR.201500224 VL - 59 IS - 12 SP - 2432-2447 J2 - Mol. Nutr. Food Res. LA - en OP - SN - 1613-4125 UR - http://dx.doi.org/10.1002/MNFR.201500224 DB - Crossref KW - Anthocyanins KW - Brain distribution KW - Catechins KW - Pharmacokinetics KW - Polyphenols ER - TY - JOUR TI - Impact of Federal, State, and Center Policies and Regulations on Nutrition Education in North Carolina Head Start Preschools AU - Peterson, Amanda D. AU - Willden, S. AU - Goodell, L.S. AU - Carraway-Stage, V. T2 - Journal of Nutrition Education and Behavior AB - The purpose of this study was to explore federal, state, and center-level policies and regulations and their affect on nutrition education in North Carolina-based Head Start preschools. Researchers conducted 63 semi-structured phone interviews with North Carolina Head Start teachers (n=32) and Health/Nutrition Coordinators (n=31). All interviews were transcribed and coded for emergent themes. Interrelated themes were condensed and aligned within a substantive-level grounded theory model. Researchers triangulated the findings by reading federal and state policies capable of impacting nutrition education; triangulation helped researchers distinguish between the actual policies versus participant perceived policies. Researchers identified four primary interrelated themes that impacted teachers ability to teach nutrition education including: classroom-based factors (e.g. federal education policies, state sanitation policies, teacher- and child-related factors), mealtime-related factors (e.g. federal mealtime requirements, quality of meals served, mealtime education), administrative-related factors (e.g. administrative priority, oversight), and policy perceptions (e.g. job-related responsibilities, policy interpretations, policy changes). Findings indicated that the provision of quality nutrition education is impacted by a multitude of policies and regulations; state-and center-level policies posed the greatest barriers for teachers and administrators. For instance, some teachers could not follow federal recommendations of educating children through hands-on food activities because of local sanitation and safety regulations. Despite policies being in place to facilitate education, development, and child safety, the cumulative effect of multiple policies and regulations may impose unintended limitations on teacher’s ability to provide quality nutrition education to preschool children. Future research is needed to explore the origin and impacts of center-level policies on nutrition education. DA - 2015/7// PY - 2015/7// DO - 10.1016/J.JNEB.2015.04.177 VL - 47 IS - 4 SP - S67 J2 - Journal of Nutrition Education and Behavior LA - en OP - SN - 1499-4046 UR - http://dx.doi.org/10.1016/J.JNEB.2015.04.177 DB - Crossref ER - TY - JOUR TI - Development of a Family Focused Child Obesity Prevention Program AU - Hughes, Sheryl AU - Power, T. AU - Johnson, S. AU - Parker, L. AU - Beck, A. AU - Overath, I. AU - Betz, D. AU - Goodell, L.S. AU - Lanigan, J. T2 - Journal of Nutrition Education and Behavior AB - The objective was to use coded data from direct observation of low-income family meals and information from focus groups to inform the development of curriculum for a seven week family intervention program. Parent, child, and family sessions were developed for African-American and Hispanic low-income families with preschoolers. Parent sessions included: Introduction, Trying New Foods (how do children develop preferences), Internal Cues of Hunger and Fullness (how do children decide when they’ve had enough to eat; how do parents know when their child has had enough to eat), Portion Sizes (how do parents decide what is an appropriate portion for preschoolers) Structure of the Environment (outside influences), Structure of the Home Environment (structure of the home meal), and Review (responsive feeding). Extensive curriculum was developed for each of the seven parent sessions and included fun and engaging videos incorporated into the content. The seven child sessions included activities targeting trying new foods and identifying internal cues similar to the content in the parent sessions. Fun and engaging dolls were crafted to help teach children to identify and describe cues of hunger and fullness. Family sessions were developed as well with the parents and children focusing on what they learned in their individual sessions. Programs are being implemented in the states of Washington and Texas by trained parent and child facilitators. A total of 6 to 8 parent-child dyads are enrolled in each intervention arm and 6-8 parent-child dyads are enrolled in each control arm receiving no intervention. Recruiting from Head Start or low-income nutrition programs in each state, five programs have been completed to date with a total of 79 parent-child dyads having completed the program (42 in intervention groups and 37 in control groups). Programs will continue in each state for the coming year and possibly into the following year. DA - 2015/7// PY - 2015/7// DO - 10.1016/J.JNEB.2015.04.294 VL - 47 IS - 4 SP - S106 J2 - Journal of Nutrition Education and Behavior LA - en OP - SN - 1499-4046 UR - http://dx.doi.org/10.1016/J.JNEB.2015.04.294 DB - Crossref ER - TY - JOUR TI - Effect of cook method on consumer perception of bacon AU - Tennant, T. AU - Drake, M.A. AU - Hanson, D.J. T2 - Meat Science DA - 2015/3// PY - 2015/3// DO - 10.1016/J.MEATSCI.2014.09.023 VL - 101 SP - 106-107 J2 - Meat Science LA - en OP - SN - 0309-1740 UR - http://dx.doi.org/10.1016/J.MEATSCI.2014.09.023 DB - Crossref ER - TY - JOUR TI - Anthocyanins and their physiologically relevant metabolites alter the expression of IL-6 and VCAM-1 in CD40L and oxidized LDL challenged vascular endothelial cells AU - Amin, Hiren P. AU - Czank, Charles AU - Raheem, Saki AU - Zhang, Qingzhi AU - Botting, Nigel P. AU - Cassidy, Aedín AU - Kay, Colin D. T2 - Molecular Nutrition & Food Research AB - Scope In vitro and in vivo studies suggest that dietary anthocyanins modulate cardiovascular disease risk; however, given anthocyanins extensive metabolism, it is likely that their degradation products and conjugated metabolites are responsible for this reported bioactivity. Methods and results Human vascular endothelial cells were stimulated with either oxidized LDL (oxLDL) or cluster of differentiation 40 ligand (CD40L) and cotreated with cyanidin‐3‐glucoside and 11 of its recently identified metabolites, at 0.1, 1, and 10 μM concentrations. Protein and gene expression of IL‐6 and VCAM‐1 was quantified by ELISA and RT‐qPCR. In oxLDL‐stimulated cells the parent anthocyanin had no effect on IL‐6 production, whereas numerous anthocyanin metabolites significantly reduced IL‐6 protein levels; phase II conjugates of protocatechuic acid produced the greatest effects (>75% reduction, p ≤ 0.05). In CD40L‐stimulated cells the anthocyanin and its phase II metabolites reduced IL‐6 protein production, where protocatechuic acid‐4‐sulfate induced the greatest reduction (>96% reduction, p ≤ 0.03). Similarly, the anthocyanin and its metabolites reduced VCAM‐1 protein production, with ferulic acid producing the greatest effect (>65% reduction, p ≤ 0.04). Conclusion These novel data provide evidence to suggest that anthocyanin metabolites are bioactive at physiologically relevant concentrations and have the potential to modulate cardiovascular disease progression by altering the expression of inflammatory mediators. DA - 2015/4/30/ PY - 2015/4/30/ DO - 10.1002/MNFR.201400803 VL - 59 IS - 6 SP - 1095-1106 J2 - Mol. Nutr. Food Res. LA - en OP - SN - 1613-4125 UR - http://dx.doi.org/10.1002/MNFR.201400803 DB - Crossref KW - Adhesion molecule KW - Cytokine KW - Cyanidin-3-glucoside KW - Inflammation KW - Metabolites ER - TY - JOUR TI - Flavonoid metabolites reduce tumor necrosis factor-α secretion to a greater extent than their precursor compounds in human THP-1 monocytes AU - di Gesso, Jessica L. AU - Kerr, Jason S. AU - Zhang, Qingzhi AU - Raheem, Saki AU - Yalamanchili, Sai Krishna AU - O'Hagan, David AU - Kay, Colin D. AU - O'Connell, Maria A. T2 - Molecular Nutrition & Food Research AB - Scope Flavonoids are generally studied in vitro, in isolation, and as unmetabolized precursor structures. However, in the habitual diet, multiple flavonoids are consumed together and found present in the circulation as complex mixtures of metabolites. Using a unique study design, we investigated the potential for singular or additive anti‐inflammatory effects of flavonoid metabolites relative to their precursor structures. Methods and results Six flavonoids, 14 flavonoid metabolites, and 29 combinations of flavonoids and their metabolites (0.1–10 μM) were screened for their ability to reduce LPS‐induced tumor necrosis factor‐α (TNF‐α) secretion in THP‐1 monocytes. One micromolar peonidin‐3‐glucoside, cyanidin‐3‐glucoside, and the metabolites isovanillic acid (IVA), IVA‐glucuronide, vanillic acid‐glucuronide, protocatechuic acid‐3‐sulfate, and benzoic acid‐sulfate significantly reduced TNF‐α secretion when in isolation, while there was no effect on TNF‐α mRNA expression. Four combinations of metabolites that included 4‐hydroxybenzoic acid (4HBA) and/or protocatechuic acid also significantly reduced TNF‐α secretion to a greater extent than the precursors or metabolites alone. The effects on LPS‐induced IL‐1β and IL‐10 secretion and mRNA expression were also examined. 4HBA significantly reduced IL‐1β secretion but none of the flavonoids or metabolites significantly modified IL‐10 secretion. Conclusion This study provides novel evidence suggesting flavonoid bioactivity results from cumulative or additive effects of circulating metabolites. DA - 2015/5/3/ PY - 2015/5/3/ DO - 10.1002/mnfr.201400799 VL - 59 IS - 6 SP - 1143-1154 J2 - Mol. Nutr. Food Res. LA - en OP - SN - 1613-4125 UR - http://dx.doi.org/10.1002/mnfr.201400799 DB - Crossref KW - Cytokine KW - Inflammation KW - Metabolism KW - Phase 2 conjugates KW - Polyphenol ER - TY - JOUR TI - Rethinking paradigms for studying mechanisms of action of plant bioactives AU - Kay, C. D. T2 - Nutrition Bulletin AB - Many foods in our diets such as berries, tea, chocolate and wine contain flavonoids, which are natural components of plants. A substantial body of evidence supports the role of flavonoids in providing protection against cardio-metabolic diseases and disorders. Despite the nearly exponential growth in flavonoid research in the past 20 years, limited progress has been made in understanding how these dietary components work. Research initially focused on their antioxidant activity without taking into account their metabolism, which now appears extensive. This has provided a new research impetus to understand the biological activity of the flavonoid metabolites. Here, we outline recent research, which suggests a highly complex interplay between metabolism, intestinal microflora, the immune system and various tissues of our body. DA - 2015/11/18/ PY - 2015/11/18/ DO - 10.1111/nbu.12178 VL - 40 IS - 4 SP - 335-339 J2 - Nutr Bull LA - en OP - SN - 1471-9827 UR - http://dx.doi.org/10.1111/nbu.12178 DB - Crossref KW - anti-inflammatory KW - bioactivity flavonoid KW - metabolism KW - microflora KW - vascular ER - TY - JOUR TI - An updated evolutionary classification of CRISPR–Cas systems AU - Makarova, Kira S. AU - Wolf, Yuri I. AU - Alkhnbashi, Omer S. AU - Costa, Fabrizio AU - Shah, Shiraz A. AU - Saunders, Sita J. AU - Barrangou, Rodolphe AU - Brouns, Stan J. J. AU - Charpentier, Emmanuelle AU - Haft, Daniel H. AU - Horvath, Philippe AU - Moineau, Sylvain AU - Mojica, Francisco J. M. AU - Terns, Rebecca M. AU - Terns, Michael P. AU - White, Malcolm F. AU - Yakunin, Alexander F. AU - Garrett, Roger A. AU - van der Oost, John AU - Backofen, Rolf AU - Koonin, Eugene V. T2 - Nature Reviews Microbiology AB - CRISPR–Cas systems provide bacteria and archaea with adaptive immunity to invading foreign DNA. In an Analysis article, Koonin and colleagues update a previous classification of these systems to incorporate the large volume of genomic data generated in recent years. The evolution of CRISPR–cas loci, which encode adaptive immune systems in archaea and bacteria, involves rapid changes, in particular numerous rearrangements of the locus architecture and horizontal transfer of complete loci or individual modules. These dynamics complicate straightforward phylogenetic classification, but here we present an approach combining the analysis of signature protein families and features of the architecture of cas loci that unambiguously partitions most CRISPR–cas loci into distinct classes, types and subtypes. The new classification retains the overall structure of the previous version but is expanded to now encompass two classes, five types and 16 subtypes. The relative stability of the classification suggests that the most prevalent variants of CRISPR–Cas systems are already known. However, the existence of rare, currently unclassifiable variants implies that additional types and subtypes remain to be characterized. DA - 2015/9/28/ PY - 2015/9/28/ DO - 10.1038/nrmicro3569 VL - 13 IS - 11 SP - 722-736 J2 - Nat Rev Microbiol LA - en OP - SN - 1740-1526 1740-1534 UR - http://dx.doi.org/10.1038/NRMICRO3569 DB - Crossref ER - TY - JOUR TI - Conserved S-Layer-Associated Proteins Revealed by Exoproteomic Survey of S-Layer-Forming Lactobacilli AU - Johnson, Brant R. AU - Hymes, Jeffrey AU - Sanozky-Dawes, Rosemary AU - Henriksen, Emily DeCrescenzo AU - Barrangou, Rodolphe AU - Klaenhammer, Todd R. T2 - Applied and Environmental Microbiology AB - The Lactobacillus acidophilus homology group comprises Gram-positive species that include L. acidophilus, L. helveticus, L. crispatus, L. amylovorus, L. gallinarum, L. delbrueckii subsp. bulgaricus, L. gasseri, and L. johnsonii. While these bacteria are closely related, they have varied ecological lifestyles as dairy and food fermenters, allochthonous probiotics, or autochthonous commensals of the host gastrointestinal tract. Bacterial cell surface components play a critical role in the molecular dialogue between bacteria and interaction signaling with the intestinal mucosa. Notably, the L. acidophilus complex is distinguished in two clades by the presence or absence of S-layers, which are semiporous crystalline arrays of self-assembling proteinaceous subunits found as the outermost layer of the bacterial cell wall. In this study, S-layer-associated proteins (SLAPs) in the exoproteomes of various S-layer-forming Lactobacillus species were proteomically identified, genomically compared, and transcriptionally analyzed. Four gene regions encoding six putative SLAPs were conserved in the S-layer-forming Lactobacillus species but not identified in the extracts of the closely related progenitor, L. delbrueckii subsp. bulgaricus, which does not produce an S-layer. Therefore, the presence or absence of an S-layer has a clear impact on the exoproteomic composition of Lactobacillus species. This proteomic complexity and differences in the cell surface properties between S-layer- and non-S-layer-forming lactobacilli reveal the potential for SLAPs to mediate intimate probiotic interactions and signaling with the host intestinal mucosa. DA - 2015/10// PY - 2015/10// DO - 10.1128/aem.01968-15 VL - 82 IS - 1 SP - 134-145 ER - TY - JOUR TI - Metagenomic reconstructions of bacterial CRISPR loci constrain population histories AU - Sun, Christine L AU - Thomas, Brian C AU - Barrangou, Rodolphe AU - Banfield, Jillian F T2 - The ISME Journal AB - Bacterial CRISPR-Cas systems provide insight into recent population history because they rapidly incorporate, in a unidirectional manner, short fragments (spacers) from coexisting infective virus populations into host chromosomes. Immunity is achieved by sequence identity between transcripts of spacers and their targets. Here, we used metagenomics to study the stability and dynamics of the type I-E CRISPR-Cas locus of Leptospirillum group II bacteria in biofilms sampled over 5 years from an acid mine drainage (AMD) system. Despite recovery of 452,686 spacers from CRISPR amplicons and metagenomic data, rarefaction curves of spacers show no saturation. The vast repertoire of spacers is attributed to phage/plasmid population diversity and retention of old spacers, despite rapid evolution of the targeted phage/plasmid genome regions (proto-spacers). The oldest spacers (spacers found at the trailer end) are conserved for at least 5 years, and 12% of these retain perfect or near-perfect matches to proto-spacer targets. The majority of proto-spacer regions contain an AAG proto-spacer adjacent motif (PAM). Spacers throughout the locus target the same phage population (AMDV1), but there are blocks of consecutive spacers without AMDV1 target sequences. Results suggest long-term coexistence of Leptospirillum with AMDV1 and periods when AMDV1 was less dominant. Metagenomics can be applied to millions of cells in a single sample to provide an extremely large spacer inventory, allow identification of phage/plasmids and enable analysis of previous phage/plasmid exposure. Thus, this approach can provide insights into prior bacterial environment and genetic interplay between hosts and their viruses. DA - 2015/9// PY - 2015/9// DO - 10.1038/ismej.2015.162 VL - 10 IS - 4 SP - 858-870 ER - TY - JOUR TI - CRISPR-Based Technologies and the Future of Food Science AU - Selle, Kurt AU - Barrangou, Rodolphe T2 - Journal of Food Science AB - The on-going CRISPR craze is focused on the use of Cas9-based technologies for genome editing applications in eukaryotes, with high potential for translational medicine and next-generation gene therapy. Nevertheless, CRISPR-Cas systems actually provide adaptive immunity in bacteria, and have much promise for various applications in food bacteria that include high-resolution typing of pathogens, vaccination of starter cultures against phages, and the genesis of programmable and specific antibiotics that can selectively modulate bacterial population composition. Indeed, the molecular machinery from these DNA-encoded, RNA-mediated, DNA-targeting systems can be harnessed in native hosts, or repurposed in engineered systems for a plethora of applications that can be implemented in all organisms relevant to the food chain, including agricultural crops trait-enhancement, livestock breeding, and fermentation-based manufacturing, and for the genesis of next-generation food products with enhanced quality and health-promoting functionalities. CRISPR-based applications are now poised to revolutionize many fields within food science, from farm to fork. In this review, we describe CRISPR-Cas systems and highlight their potential for the development of enhanced foods. DA - 2015/10/7/ PY - 2015/10/7/ DO - 10.1111/1750-3841.13094 VL - 80 IS - 11 SP - R2367-R2372 KW - CRISPR KW - genome KW - microbiology ER - TY - JOUR TI - Diversity of CRISPR-Cas immune systems and molecular machines AU - Barrangou, Rodolphe T2 - Genome Biology AB - Bacterial adaptive immunity hinges on CRISPR-Cas systems that provide DNA-encoded, RNA-mediated targeting of exogenous nucleic acids. A plethora of CRISPR molecular machines occur broadly in prokaryotic genomes, with a diversity of Cas nucleases that can be repurposed for various applications. DA - 2015/11/9/ PY - 2015/11/9/ DO - 10.1186/s13059-015-0816-9 VL - 16 IS - 1 ER - TY - JOUR TI - Orange juice--derived flavanone and phenolic metabolites do not acutely affect cardiovascular risk biomarkers: a randomized, placebo-controlled, crossover trial in men at moderate risk of cardiovascular disease-- T2 - The American journal of clinical nutrition DA - 2015/// PY - 2015/// VL - 101 IS - 5 SP - 931-938 ER - TY - JOUR TI - Phenolic Metabolites of Anthocyanins Modulate Mechanisms of Endothelial Function AU - Edwards, Michael AU - Czank, Charles AU - Woodward, Gary M. AU - Cassidy, Aedín AU - Kay, Colin D. T2 - Journal of Agricultural and Food Chemistry AB - Anthocyanins are reported to have vascular bioactivity, however their mechanisms of action are largely unknown. Evidence suggests that anthocyanins modulate endothelial function, potentially by increasing nitric oxide (NO) synthesis, or enhancing NO bioavailability. This study compared the activity of cyanidin-3-glucoside, its degradation product protocatechuic acid, and phase II metabolite, vanillic acid. Production of NO and superoxide and expression of endothelial NO synthase (eNOS), NADPH oxidase (NOX), and heme oxygenase-1 (HO-1) were established in human vascular cell models. Nitric oxide levels were not modulated by the treatments, although eNOS was upregulated by cyanidin-3-glucoside, and superoxide production was decreased by both phenolic acids. Vanillic acid upregulated p22(phox) mRNA but did not alter NOX protein expression, although trends were observed for p47(phox) downregulation and HO-1 upregulation. Anthocyanin metabolites may therefore modulate vascular reactivity by inducing HO-1 and modulating NOX activity, resulting in reduced superoxide production and improved NO bioavailability. DA - 2015/3/2/ PY - 2015/3/2/ DO - 10.1021/JF5041993 VL - 63 IS - 9 SP - 2423-2431 J2 - J. Agric. Food Chem. LA - en OP - SN - 0021-8561 1520-5118 UR - http://dx.doi.org/10.1021/JF5041993 DB - Crossref KW - endothelium KW - eNOS KW - cyanidin KW - HUVEC KW - NADPH-oxidase ER - TY - JOUR TI - Anthocyanins and their physiologically relevant metabolites alter the expression of IL-6 and VCAM-1 in CD40L and oxidized LDL challenged vascular endothelial cells T2 - Molecular nutrition & food research DA - 2015/// PY - 2015/// VL - 59 IS - 6 SP - 1095-1106 ER - TY - JOUR TI - Neuroprotective effects of dried camu-camu (Myrciaria dubia HBK McVaugh) residue in C. elegans AU - Azevêdo, Juliana C.S. AU - Borges, Kátia C. AU - Genovese, Maria I. AU - Correia, Roberta T.P. AU - Vattem, Dhiraj A. T2 - Food Research International AB - The effect of hot air dried camu-camu (Myrciaria dubia HBK McVaugh) residue on modulating redox response signaling was evaluated in transgenic Caenorhabditis elegans. Camu-camu residue was fractionated into low and high molecular weight fractions and used as treatments. Relative fold changes in gene expression in response to camu-camu treatments were quantified using fluorescence microscopy. Also, the neuroprotective effects of camu-camu residue in experimentally induced neurodegeneration were evaluated in C. elegans models for Alzheimer's disease (AD) and Parkinson's disease (PD). For AD, time to thermally induced Aβ1–42 aggregation mediated paralysis was evaluated in transgenic C. elegans (CL4176). For PD, MPP + induced neurodegeneration was quantified by loss in motility due to paralysis. Results suggest a significant upregulation expression of superoxide dismutase (SOD-3 and SOD-4) and catalases (CTL-1; CTL-2; CTL-3) in response to treatment with camu-camu residue, especially with the low molecular weight fraction. Furthermore, treatment with this fraction significantly extended the life span in C. elegans by 20% and delayed Aβ1–42 induced paralysis by 21%. Additionally, treatment with camu-camu residue also abrogated MPP + induced neurodegeneration for PD by 15–21%. DA - 2015/7// PY - 2015/7// DO - 10.1016/j.foodres.2015.02.015 UR - https://doi.org/10.1016/j.foodres.2015.02.015 KW - Camu-camu KW - Neurodegeneration KW - Life span KW - Alzheimer's disease KW - Parkinson's disease ER - TY - JOUR TI - Caprine frozen yoghurt produced with fresh and spray dried jambolan fruit pulp (Eugenia jambolana Lam) and Bifidobacterium animalis subsp. lactis BI-07 AU - Bezerra, M. AU - Araujo, A. AU - Santos, K. AU - Correia, R. T2 - LWT - Food Science and Technology AB - The present study investigated four experimental groups of caprine frozen yoghurt produced with the addition of fresh and spray dried jambolan fruit pulp (Eugenia jambolana Lam) and the probiotic strain Bifidobacterium animalis subsp. lactis BI-07. The physicochemical characteristics, overrun, meltdown test, selected bioactive-related attributes, besides the survival of added probiotic bacteria during the frozen storage and the sensory acceptance of the products were investigated. The probiotic addition decreased the pH of frozen yoghurt samples (P < 0.05) and low overrun values (14.2–22.6%) were observed for all samples. Higher phenolic content for samples with the probiotic strain was observed (P < 0.05), which suggests a possible biotransformation that would lead to enhanced phytochemical level. The frozen yoghurt samples with probiotic presented high initial cell counts (approx. 9 log CFU/g) and an outstanding probiotic survival rate (97%) was observed throughout the 90 days of frozen storage. Frozen yoghurt samples with probiotic bacteria received significantly lower flavor scores (P < 0.05), and similar results were obtained for appearance, aroma and consistency among frozen yoghurt samples. Overall, the caprine frozen yoghurt enriched with jambolan fruit proved to be an efficient carrier for B. animalis subsp. lactis BI-07 and presented desirable bioactive and technological attributes. DA - 2015/7// PY - 2015/7// DO - 10.1016/j.lwt.2015.01.049 VL - 62 IS - 2 SP - 1099-1104 UR - https://doi.org/10.1016/j.lwt.2015.01.049 KW - Goat milk KW - Dairy KW - Bioactive compounds KW - Probiotic ER - TY - JOUR TI - Potentially probiotic ice cream from goat's milk: Characterization and cell viability during processing, storage and simulated gastrointestinal conditions AU - Silva, Priscilla Diniz Lima AU - Fátima Bezerra, Maria AU - Santos, Karina Maria Olbrich AU - Correia, Roberta Targino Pinto T2 - LWT - Food Science and Technology AB - In this work, the physicochemical characteristics, meltdown behavior and sensory properties of goat's milk ice cream produced with and without the probiotic bacteria Bifidobacterium animalis subsp. lactis BLC1 were analyzed. The ice cream with added B. animalis was further evaluated in regard to the probiotic viability during processing, frozen storage, and simulated gastrointestinal conditions. Results showed that the addition of B. animalis decreased the pH (p < 0.05), but it had no effect on physicochemical properties, including overrun and melting behavior of ice cream from goat's milk (p > 0.05). After 120 days of frozen storage, a survival rate of 84.7% was registered. With regard to cell viability during gastrointestinal conditions, the exposure to bile and pancreatin resulted in the decline of 3.82 log cycles in ice cream samples previously stored at −18 °C for 120 days. Overall, the goat's milk ice cream with B. animalis received good sensory scores and satisfactory probiotic viability (6–7 log CFU/g) was maintained throughout the 120 days of frozen storage. Therefore, this research shows that goat's milk ice cream is an adequate delivery vehicle for the probiotic bacteria B. animalis. DA - 2015/6// PY - 2015/6// DO - 10.1016/j.lwt.2014.02.055 UR - https://doi.org/10.1016/j.lwt.2014.02.055 KW - Goat's milk KW - Ice cream KW - Probiotic survival KW - Gastrointestinal simulation ER - TY - JOUR TI - High-Pressure Enhanced Infusion: Influence of Process Parameters AU - Mahadevan, S. AU - Nitin, N. AU - Salvi, D. AU - Karwe, M.V. T2 - Journal of Food Process Engineering AB - Abstract High‐pressure processing ( HPP ) has been shown to enhance infusion of small‐size molecules into foods primarily because of cell membrane rupture. The enhanced infusion has been previously explained using increased effective diffusivity, which does not elucidate the actual mass transport mechanisms. This study evaluates and explains the role of transport processes and the influence of process parameters on enhanced infusion of antioxidants by HPP . Frozen‐thawed cranberries were infused with quercetin. A greater amount of quercetin (three times) was infused faster (18 times) in cranberries after HPP (100 MPa–551 MP a at 22 C ) compared with those infused at ambient conditions. While freeze‐thawing process ruptured cell membranes in cranberries, measured as cell permeability, there was no additional cell rupture after HPP . A similar degree of measured cell permeability before (0.48 ± 0.01) and after HPP (0.48 ± 0.04), and observed enhanced infusion under high‐pressure suggest that cell membrane permeabilization may not be the only cause for high–pressure‐assisted infusion. Practical Applications Conventional diffusion‐based mass transport methods currently used in the food industry are slow processes. In order to accelerate the process, this research investigates (1) the potential of high‐pressure processing ( HPP ) to enhance and accelerate infusion in cranberries and (2) the mechanisms operative during high–pressure‐assisted infusion. The present study was focused on understanding influence of process variables on extent of infusion and cell permeability. HPP significantly enhanced and accelerated infusion of quercetin in cranberries. Additionally, the commonly accepted explanation of cell permeabilization being the only cause for enhanced infusion under high pressure was not found to be operative in the current system of study. The success of this research can highlight the potential of HPP as an alternative technology that will be useful to the food industry to develop a broad class of nutrient infused food products. DA - 2015/// PY - 2015/// DO - 10.1111/jfpe.12190 VL - 38 IS - 6 SP - 601-612 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-84944169510&partnerID=MN8TOARS ER - TY - JOUR TI - High pressure-enhanced infusion in fresh and frozen-thawed cranberries: A comparative study AU - Mahadevan, S. AU - Salvi, D. AU - Karwe, M.V. T2 - Journal of Food Process Engineering AB - Abstract High‐pressure processing ( HPP ) has been shown to enhance and accelerate infusion of molecules compared with osmosis‐driven diffusion. The primary cause for enhanced HPP infusion is attributed to cell membrane permeabilization. This study tests the validity of this commonly accepted mechanism by conducting HPP (100 MPa–551 MPa) infusion of quercetin into cranberries. Two systems used in this study were fresh cranberries, cells of which are intact, and frozen‐thawed cranberries, cells of which are permeabilized during freeze‐thaw process. Results showed that while fresh and frozen‐thawed cranberries had similar cell membrane permeability after HPP, twice as much quercetin was infused into fresh cranberries as compared with frozen‐thawed cranberries. If cell membrane permeability were the only cause of infusion, the amount of quercetin infused into cranberries in both systems should have been same. These results suggest that enhanced infusion caused by HPP is not just caused by cell permeabilization alone. Practical Applications Conventional diffusion based mass transport methods, currently used in the food industry for infusion, are slow processes. In order to accelerate the mass transport, this research investigated the potential of HPP to enhance and accelerate infusion in cranberries. The present study was focused on the comparison of infusion during HPP using fresh cranberries, cells of which were intact and frozen‐thawed cranberries, cells of which were permeabilized (with ruptured cell membranes) by freeze‐thaw process. Results showing increased infusion in fresh cranberries and similar cell permeability in both fresh and frozen‐thawed cranberries suggested that enhanced infusion during HPP was not caused only by cell permeabilization. This research opens new avenues for investigating additional mass transport mechanisms occurring during HPP , which will help develop accurate predictive mass transport models for HPP infusion processes. DA - 2015/// PY - 2015/// DO - 10.1111/jfpe.12198 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-84922880226&partnerID=MN8TOARS ER - TY - JOUR TI - Effect of temperature abuse on frozen army rations. Part 1: Developing a heat transfer numerical model based on thermo-physical properties of food AU - Karthikeyan, J.S. AU - Desai, K.M. AU - Salvi, D. AU - Bruins, R. AU - Karwe, M.V. T2 - Food Research International AB - Numerical simulation was carried out to predict the effect of external temperature conditions on thermal behavior of frozen US military rations, during storage and transportation. An army breakfast menu box containing beefsteaks, concentrated orange juice, peppers & onions, French toast, and Danishes, was selected for conducting this study. Thermo-physical properties of each food item were characterized using their composition and differential scanning calorimeter (DSC). Apparent heat capacity method was used to account for the latent heat of phase change during simulation of thawing and freezing. Numerically simulated results were experimentally validated using a gel-based model food system and the food items in the menu box. The average deviation between numerically predicted temperature and experimentally measured temperature for the model food system was approximately 1 °C and for the targeted food items the deviation ranged from 2 °C to 5 °C, depending on the food item. DA - 2015/// PY - 2015/// DO - 10.1016/j.foodres.2015.07.007 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-84937676028&partnerID=MN8TOARS KW - Numerical simulation KW - Frozen food KW - Differential scanning calorimeter KW - Apparent heat capacity ER - TY - JOUR TI - Effect of temperature abuse on frozen army rations. Part 2: Predicting microbial spoilage AU - Karthikeyan, J.S. AU - Desai, K.M. AU - Salvi, D. AU - Bruins, R. AU - Schaffner, D.W. AU - Karwe, M.V. T2 - Food Research International AB - Numerically simulated heat transfer model of frozen US military rations was combined with microbial kinetics to predict the microbial spoilage of the food products, during two possible temperature abuse scenarios. An army breakfast menu box containing five different food items was selected for conducting this research. One of the food item in the menu box, beefsteak, was chosen for detailed microbial study. A microbial predictive tool was used to identify and evaluate the kinetics of the most prone microorganism that can grow in a beefsteak. Numerical predictions suggested that the food items exposed to external temperatures ranging from 20°C to 40°C can be allowed to stay at those temperatures for maximum times of 28.7h to 11.9h, respectively. The food items can be allowed to stay inside the broken freezer for a maximum time of 186h, to ensure microbial safety in the case of freezer failure. DA - 2015/// PY - 2015/// DO - 10.1016/j.foodres.2015.07.012 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-84937468736&partnerID=MN8TOARS KW - Numerical simulation KW - Frozen food KW - Temperature abuse KW - Microbial spoilage ER - TY - JOUR TI - In-Feed Supplementation of trans-Cinnamaldehyde Reduces Layer-Chicken Egg-Borne Transmission of Salmonella enterica Serovar Enteritidis AU - Upadhyaya, Indu AU - Upadhyay, Abhinav AU - Kollanoor-Johny, Anup AU - Mooyottu, Shankumar AU - Baskaran, Sangeetha A. AU - Yin, Hsin-Bai AU - Schreiber, David T. AU - Khan, Mazhar I. AU - Darre, Michael J. AU - Curtis, Patricia A. AU - Venkitanarayanan, Kumar T2 - APPLIED AND ENVIRONMENTAL MICROBIOLOGY AB - ABSTRACT Salmonella enterica serovar Enteritidis is a major foodborne pathogen in the United States, causing gastroenteritis in humans, primarily through consumption of contaminated eggs. Chickens are the reservoir host of S . Enteritidis. In layer hens, S . Enteritidis colonizes the intestine and migrates to various organs, including the oviduct, leading to egg contamination. This study investigated the efficacy of in-feed supplementation with trans -cinnamaldehyde (TC), a generally recognized as safe (GRAS) plant compound obtained from cinnamon, in reducing S . Enteritidis cecal colonization and systemic spread in layers. Additionally, the effect of TC on S . Enteritidis virulence factors critical for macrophage survival and oviduct colonization was investigated in vitro . The consumer acceptability of eggs was also determined by a triangle test. Supplementation of TC in feed for 66 days at 1 or 1.5% (vol/wt) for 40- or 25-week-old layer chickens decreased the amounts of S . Enteritidis on eggshell and in yolk ( P < 0.001). Additionally, S . Enteritidis persistence in the cecum, liver, and oviduct in TC-supplemented birds was decreased compared to that in controls ( P < 0.001). No significant differences in feed intake, body weight, or egg production in birds or in consumer acceptability of eggs were observed ( P > 0.05). In vitro cell culture assays revealed that TC reduced S . Enteritidis adhesion to and invasion of primary chicken oviduct epithelial cells and reduced S . Enteritidis survival in chicken macrophages ( P < 0.001). Follow-up gene expression analysis using real-time quantitative PCR (qPCR) showed that TC downregulated the expression of S . Enteritidis virulence genes critical for chicken oviduct colonization ( P < 0.001). The results suggest that TC may potentially be used as a feed additive to reduce egg-borne transmission of S . Enteritidis. DA - 2015/5// PY - 2015/5// DO - 10.1128/aem.03809-14 VL - 81 IS - 9 SP - 2985-2994 SN - 1098-5336 ER - TY - JOUR TI - Evaluation of US Poison Center Data for Surveillance of Foodborne Disease AU - Gruber, Joann F. AU - Bailey, Jennifer E. AU - Kowalcyk, Barbara B. T2 - FOODBORNE PATHOGENS AND DISEASE AB - Enhancing foodborne disease (FBD) surveillance and improving the timeliness of outbreak detection have been identified as public health priorities. Consumer complaint data have become increasingly useful for FBD surveillance and the detection of outbreaks. Calls to poison centers are a potential source of consumer complaint data. A retrospective analysis of data from the National Poison Data System (NPDS) (2000-2011) was undertaken to evaluate the value of data collected through the United States poison centers for detection of large national outbreaks and recalls.Demographic and clinical data were summarized. Prevalences of FBD calls were calculated and analyzed for time trends. Significant increases in daily call prevalences were identified, and dates of the increases were compared to the announcement of 18 national outbreaks/recalls.Over the 12-year period, there were 433,788 unique calls self-reporting a suspected FBD exposure in humans. Overall, daily call prevalences decreased over time. Only about half of callers reported common gastrointestinal clinical effects. Of the 42 identified significant increases in call prevalences, none occurred within 14 days before an outbreak announcement; 7 occurred within 14 days after an outbreak announcement.Based on this analysis, there are significant limitations to using self-reported FBD exposures to NPDS as a source of information for FBD surveillance of large national outbreaks and recalls; however, a syndromic approach may yield different results and should be explored. Improved data collection and coordination with public health agencies may improve the ability to use NPDS data to monitor FBD in near real-time, identify potential outbreaks, and improve situational awareness. DA - 2015/6/1/ PY - 2015/6/1/ DO - 10.1089/fpd.2014.1907 VL - 12 IS - 6 SP - 467-478 SN - 1556-7125 ER - TY - JOUR TI - Application of Choice Experiments to Determine Stakeholder Preferences for Woody Biomass Harvesting Guidelines AU - Serenari, Christopher AU - Peterson, M. Nils AU - Moorman, Christopher E. AU - Cubbage, Frederick AU - Jervis, Suzanne T2 - JOURNAL OF SUSTAINABLE FORESTRY AB - Biomass harvesting guidelines (BHGs) have been developed to address concerns about the sustainability of harvesting woody biomass. Assessing preferences among BHG stakeholders is important for designing operationally feasible and socially acceptable standards in different contexts. We used choice modeling to determine how foresters, loggers, and landowners perceived the relative importance of stumpage price, wildlife habitat quality, percentage of coarse woody debris (CWD) remaining, and distribution of CWD in their choices of BHG scenarios. Responses (N = 718) indicated stumpage price was nearly double the importance of wildlife habitat quality, and three times more important than debris distribution and debris remaining. DA - 2015/// PY - 2015/// DO - 10.1080/10549811.2015.1007511 VL - 34 IS - 4 SP - 343-357 SN - 1540-756X KW - woody biomass KW - guidelines KW - BHG KW - choice modeling KW - choice-based conjoint ER - TY - JOUR TI - Characterization of Phenolic Compounds and Antioxidant and Antiinflammatory Activities from Mamuyo (Styrax ramirezii Greenm.) Fruit AU - Timmers, Michael A. AU - Guerrero-Medina, Jorge L. AU - Esposito, Debora AU - Grace, Mary H. AU - Paredes-Lopez, Octavio AU - Garcia-Saucedo, Pedro A. AU - Ann Lila, Mary T2 - JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY AB - Extracts of Styrax ramirezii Greenm., a fruit traditionally valued for health and wellness in Mexico, were analyzed phytochemically and evaluated for antioxidant and anti-inflammatory activity. Six norneolignans were identified by HPLC-TOF-MS, and the two major compounds were isolated for further evaluation. The effects of the isolated norneolignans, egonol and homoegonol, on lipopolysaccharide (LPS)-induced nitric oxide (NO) production, reactive oxygen species (ROS) production, and biomarkers of inflammation were evaluated. Of the tested compounds, egonol potently inhibited the production of NO and also significantly reduced the release of ROS. Consistent with these observations, the mRNA expression levels of inducible nitric oxide synthase (iNOS) (0.668 ± 0.108), cyclooxygenase-2 (COX-2) (0.553 ± 0.007), interleukin-1β (IL-1β) (0.093 ± 0.005), and interleukin-6 (IL-6) (0.298 ± 0.076) were reduced by egonol. The activity for both egonol and homoegonol increased in a concentration-dependent manner. These results suggest the potential of S. ramirezii Greenm. fruit to contribute to a healthy diet, rich in antioxidant and anti-inflammatory compounds. DA - 2015/12/9/ PY - 2015/12/9/ DO - 10.1021/acs.jafc.5b04781 VL - 63 IS - 48 SP - 10459-10465 SN - 1520-5118 KW - LC-TOF-MS KW - Styrax ramirezii KW - antioxidant activity KW - anti-inflammatory activity KW - norneolignan ER - TY - JOUR TI - Mitochondrial DNA Fragmentation to Monitor Processing Parameters in High Acid, Plant-Derived Foods AU - Caldwell, Jane M. AU - Perez-Diaz, Ilenys M. AU - Harris, Keith AU - Hassan, Hosni M. AU - Simunovic, Josip AU - Sandeep, K. P. T2 - JOURNAL OF FOOD SCIENCE AB - Abstract Mitochondrial DNA (mtDNA) fragmentation was assessed in acidified foods. Using quantitative polymerase chain reaction, C t values measured from fresh, fermented, pasteurized, and stored cucumber mtDNA were determined to be significantly different ( P > 0.05) based on processing and shelf‐life. This indicated that the combination of lower temperature thermal processes (hot‐fill at 75 °C for 15 min) and acidified conditions (pH = 3.8) was sufficient to cause mtDNA fragmentation. In studies modeling high acid juices, pasteurization (96 °C, 0 to 24 min) of tomato serum produced C t values which had high correlation to time‐temperature treatment. Primers producing longer amplicons (approximately 1 kb) targeting the same mitochondrial gene gave greater sensitivity in correlating time‐temperature treatments to C t values. Lab‐scale pasteurization studies using C t values derived from the longer amplicon differentiated between heat treatments of tomato serum (95 °C for <2 min). MtDNA fragmentation was shown to be a potential new tool to characterize low temperature (<100 °C) high acid processes (pH < 4.6), nonthermal processes such as vegetable fermentation and holding times of acidified, plant‐derived products. DA - 2015/12// PY - 2015/12// DO - 10.1111/1750-3841.13139 VL - 80 IS - 12 SP - M2892-M2898 SN - 1750-3841 KW - high-acid foods KW - mitochondrial DNA KW - quantitative PCR KW - thermal processing ER - TY - JOUR TI - Food Protein Functionality-A New Model AU - Foegeding, E. Allen T2 - JOURNAL OF FOOD SCIENCE AB - Proteins in foods serve dual roles as nutrients and structural building blocks. The concept of protein functionality has historically been restricted to nonnutritive functions--such as creating emulsions, foams, and gels--but this places sole emphasis on food quality considerations and potentially overlooks modifications that may also alter nutritional quality or allergenicity. A new model is proposed that addresses the function of proteins in foods based on the length scale(s) responsible for the function. Properties such as flavor binding, color, allergenicity, and digestibility are explained based on the structure of individual molecules; placing this functionality at the nano/molecular scale. At the next higher scale, applications in foods involving gelation, emulsification, and foam formation are based on how proteins form secondary structures that are seen at the nano and microlength scales, collectively called the mesoscale. The macroscale structure represents the arrangements of molecules and mesoscale structures in a food. Macroscale properties determine overall product appearance, stability, and texture. The historical approach of comparing among proteins based on forming and stabilizing specific mesoscale structures remains valid but emphasis should be on a common means for structure formation to allow for comparisons across investigations. For applications in food products, protein functionality should start with identification of functional needs across scales. Those needs are then evaluated relative to how processing and other ingredients could alter desired molecular scale properties, or proper formation of mesoscale structures. This allows for a comprehensive approach to achieving the desired function of proteins in foods. DA - 2015/12// PY - 2015/12// DO - 10.1111/1750-3841.13116 VL - 80 IS - 12 SP - C2670-C2677 SN - 1750-3841 KW - foam KW - gel KW - protein KW - protein functionality KW - sol ER - TY - JOUR TI - Commercial Scale Cucumber Fermentations Brined with Calcium Chloride Instead of Sodium Chloride AU - Perez-Diaz, I. M. AU - McFeeters, R. F. AU - Moeller, L. AU - Johanningsmeier, S. D. AU - Hayes, J. AU - Fornea, D. S. AU - Rosenberg, L. AU - Gilbert, C. AU - Custis, N. AU - Beene, K. AU - Bass, D. T2 - JOURNAL OF FOOD SCIENCE AB - Development of low salt cucumber fermentation processes present opportunities to reduce the amount of sodium chloride (NaCl) that reaches fresh water streams from industrial activities. The objective of this research was to translate cucumber fermentation brined with calcium chloride (CaCl2 ) instead of NaCl to commercial scale production. Although CaCl2 brined cucumber fermentations were stable in laboratory experiments, commercial scale trials using 6440 L open-top tanks rapidly underwent secondary cucumber fermentation. It was understood that a limited air purging routine, use of a starter culture and addition of preservatives to the cover brine aids in achieving the desired complete cucumber fermentation. The modified process was used for subsequent commercial trials using 12490 and 28400 L open-top tanks packed with variable size cucumbers and from multiple lots, and cover brines containing CaCl2 and potassium sorbate to equilibrated concentrations of 100 and 6 mM, respectively. Lactobacillus plantarum LA0045 was inoculated to 10(6) CFU/mL, and air purging was applied for two 2-3 h periods per day for the first 10 d of fermentation and one 2-3 h period per day between days 11 and 14. All fermentations were completed, as evidenced by the full conversion of sugars to lactic acid, decrease in pH to 3.0, and presented microbiological stability for a minimum of 21 d. This CaCl2 process may be used to produce fermented cucumbers intended to be stored short term in a manner that reduces pollution and waste removal costs. DA - 2015/12// PY - 2015/12// DO - 10.1111/1750-3841.13107 VL - 80 IS - 12 SP - M2827-M2836 SN - 1750-3841 KW - sustainable processing KW - cucumber fermentation KW - low salt KW - vegetable preservation KW - chloride waste reduction ER - TY - JOUR TI - Comparison of process control viruses for use in extraction and detection of human norovirus from food matrices AU - Gentry-Shields, Jennifer AU - Jaykus, Lee-Ann T2 - FOOD RESEARCH INTERNATIONAL AB - Abstract Although RT-qPCR is a powerful tool for human norovirus (HuNoV) detection, low virus concentrations in potentially large sample volumes necessitate the use of inefficient sample processing step(s) prior to detection. Process control viruses (PCVs) are used to monitor the efficiency of these virus concentration steps. This study compared five PCVs [Mengovirus (Mengo), murine norovirus (MNV-1), MS2 coliphage, Tulane virus, and turnip crinkle virus (TCV)] to two HuNoV strains for recovery during the steps of elution, polyethylene glycol precipitation (PEG), and RNA extraction from select foods (lettuce and sliced deli ham). Results demonstrate high recovery efficiencies of HuNoV GI.6 and GII.4 using the methods described in this study: combined (sequential) losses during processing from sliced deli ham and lettuce were 10 genome equivalent copies (GEC). When considering the processing steps separately, HuNoV loss was negligible after elution, and low after PEG precipitation (mean 0.5 log 10 GEC) and RNA extraction (mean 0.1 log 10 GEC). The virus that least mimicked the behavior of HuNoV during sample processing was MNV-1. Of the viruses tested, a commercial mengovirus strain gave recovery efficiencies closest to HuNoV, showing combined losses from sliced deli ham and lettuce of 10 GEC and ~ 1 log 10 GEC, respectively. All PCVs do not behave equivalently and validation of their performance is recommended before their routine use on an application-by-application basis. DA - 2015/11// PY - 2015/11// DO - 10.1016/j.foodres.2015.05.027 VL - 77 SP - 320-325 SN - 1873-7145 KW - Norovirus KW - Process control virus KW - RT-qPCR KW - Processing efficiency ER - TY - JOUR TI - The Bacterial Origins of the CRISPR Genome-Editing Revolution AU - Sontheimer, Erik J. AU - Barrangou, Rodolphe T2 - HUMAN GENE THERAPY AB - Like most of the tools that enable modern life science research, the recent genome-editing revolution has its biological roots in the world of bacteria and archaea. Clustered, regularly interspaced, short palindromic repeats (CRISPR) loci are found in the genomes of many bacteria and most archaea, and underlie an adaptive immune system that protects the host cell against invasive nucleic acids such as viral genomes. In recent years, engineered versions of these systems have enabled efficient DNA targeting in living cells from dozens of species (including humans and other eukaryotes), and the exploitation of the resulting endogenous DNA repair pathways has provided a route to fast, easy, and affordable genome editing. In only three years after RNA-guided DNA cleavage was first harnessed, the ability to edit genomes via simple, user-defined RNA sequences has already revolutionized nearly all areas of biological science. CRISPR-based technologies are now poised to similarly revolutionize many facets of clinical medicine, and even promise to advance the long-term goal of directly editing genomic sequences of patients with inherited disease. In this review, we describe the biological and mechanistic basis for these remarkable immune systems, and how their engineered derivatives are revolutionizing basic and clinical research. DA - 2015/7/1/ PY - 2015/7/1/ DO - 10.1089/hum.2015.091 VL - 26 IS - 7 SP - 413-424 SN - 1557-7422 ER - TY - JOUR TI - Mucosal Immunogenicity of Genetically Modified Lactobacillus acidophilus Expressing an HIV-1 Epitope within the Surface Layer Protein AU - Kajikawa, Akinobu AU - Zhang, Lin AU - LaVoy, Alora AU - Bumgardner, Sara AU - Klaenhammer, Todd R. AU - Dean, Gregg A. T2 - PLOS ONE AB - Surface layer proteins of probiotic lactobacilli are theoretically efficient epitope-displaying scaffolds for oral vaccine delivery due to their high expression levels and surface localization. In this study, we constructed genetically modified Lactobacillus acidophilus strains expressing the membrane proximal external region (MPER) from human immunodeficiency virus type 1 (HIV-1) within the context of the major S-layer protein, SlpA. Intragastric immunization of mice with the recombinants induced MPER-specific and S-layer protein-specific antibodies in serum and mucosal secretions. Moreover, analysis of systemic SlpA-specific cytokines revealed that the responses appeared to be Th1 and Th17 dominant. These findings demonstrated the potential use of the Lactobacillus S-layer protein for development of oral vaccines targeting specific peptides. DA - 2015/10/28/ PY - 2015/10/28/ DO - 10.1371/journal.pone.0141713 VL - 10 IS - 10 SP - SN - 1932-6203 ER - TY - JOUR TI - Diversity of CRISPR-Cas immune systems and molecular machines AU - Barrangou, R. T2 - Genome Biology DA - 2015/// PY - 2015/// VL - 16 ER - TY - JOUR TI - Sensory and Functionality Differences of Whey Protein Isolate Bleached by Hydrogen or Benzoyl Peroxide AU - Smith, Tucker J. AU - Foegeding, E. Allen AU - Drake, MaryAnne T2 - JOURNAL OF FOOD SCIENCE AB - Whey protein is a highly functional food ingredient used in a wide variety of applications. A large portion of fluid whey produced in the United States is derived from Cheddar cheese manufacture and contains annatto (norbixin), and therefore must be bleached. The objective of this study was to compare sensory and functionality differences between whey protein isolate (WPI) bleached by benzoyl peroxide (BP) or hydrogen peroxide (HP). HP and BP bleached WPI and unbleached controls were manufactured in triplicate. Descriptive sensory analysis and gas chromatography-mass spectrometry were conducted to determine flavor differences between treatments. Functionality differences were evaluated by measurement of foam stability, protein solubility, SDS-PAGE, and effect of NaCl concentration on gelation relative to an unbleached control. HP bleached WPI had higher concentrations of lipid oxidation and sulfur containing volatile compounds than both BP and unbleached WPI (P < 0.05). HP bleached WPI was characterized by high aroma intensity, cardboard, cabbage, and fatty flavors, while BP bleached WPI was differentiated by low bitter taste. Overrun and yield stress were not different among WPI (P < 0.05). Soluble protein loss at pH 4.6 of WPI decreased by bleaching with either hydrogen peroxide or benzoyl peroxide (P < 0.05), and the heat stability of WPI was also distinct among WPI (P < 0.05). SDS PAGE results suggested that bleaching of whey with either BP or HP resulted in protein degradation, which likely contributed to functionality differences. These results demonstrate that bleaching has flavor effects as well as effects on many of the functionality characteristics of whey proteins.Whey protein isolate (WPI) is often used for its functional properties, but the effect of oxidative bleaching chemicals on the functional properties of WPI is not known. This study identifies the effects of hydrogen peroxide and benzoyl peroxide on functional and flavor characteristics of WPI bleached by hydrogen and benzoyl peroxide and provides insights for the product applications which may benefit from bleaching. DA - 2015/10// PY - 2015/10// DO - 10.1111/1750-3841.13000 VL - 80 IS - 10 SP - C2153-C2160 SN - 1750-3841 KW - bleaching KW - functionality KW - sensory KW - whey protein isolate ER - TY - JOUR TI - Occurrence and activity of a type II CRISPR-Cas system in Lactobacillus gasseri AU - Sanozky-Dawes, Rosemary AU - Selle, Kurt AU - O'Flaherty, Sarah AU - Klaenhammer, Todd AU - Barrangou, Rodolphe T2 - MICROBIOLOGY-SGM AB - Bacteria encode clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated genes (cas), which collectively form an RNA-guided adaptive immune system against invasive genetic elements. In silico surveys have revealed that lactic acid bacteria harbour a prolific and diverse set of CRISPR-Cas systems. Thus, the natural evolutionary role of CRISPR-Cas systems may be investigated in these ecologically, industrially, scientifically and medically important microbes. In this study, 17 Lactobacillus gasseri strains were investigated and 6 harboured a type II-A CRISPR-Cas system, with considerable diversity in array size and spacer content. Several of the spacers showed similarity to phage and plasmid sequences, which are typical targets of CRISPR-Cas immune systems. Aligning the protospacers facilitated inference of the protospacer adjacent motif sequence, determined to be 5'-NTAA-3' flanking the 3' end of the protospacer. The system in L. gasseri JV-V03 and NCK 1342 interfered with transforming plasmids containing sequences matching the most recently acquired CRISPR spacers in each strain. We report the distribution and function of a native type II-A CRISPR-Cas system in the commensal species L. gasseri. Collectively, these results open avenues for applications for bacteriophage protection and genome modification in L. gasseri, and contribute to the fundamental understanding of CRISPR-Cas systems in bacteria. DA - 2015/9// PY - 2015/9// DO - 10.1099/mic.0.000129 VL - 161 SP - 1752-1761 SN - 1350-0872 ER - TY - JOUR TI - Novel value-added uses for sweet potato juice and flour in polyphenol- and protein-enriched functional food ingredients AU - Grace, Mary H. AU - Truong, An N. AU - Truong, Van-Den AU - Raskin, Ilya AU - Lila, Mary Ann T2 - FOOD SCIENCE & NUTRITION AB - Abstract Blackcurrant, blueberry, and muscadine grape juices were efficiently sorbed, concentrated, and stabilized into dry granular ingredient matrices which combined anti‐inflammatory and antioxidant fruit polyphenols with sweet potato functional constituents (carotenoids, vitamins, polyphenols, fibers). Total phenolics were highest in blackcurrant‐orange sweet potato ingredient matrices (34.03 mg/g), and lowest in muscadine grape‐yellow sweet potato matrices (10.56 mg/g). Similarly, anthocyanins were most concentrated in blackcurrant‐fortified orange and yellow sweet potato matrices (5.40 and 6.54 mg/g, respectively). Alternatively, other protein‐rich edible matrices (defatted soy flour, light roasted peanut flour, and rice protein concentrate) efficiently captured polyphenols (6.09–9.46 mg/g) and anthocyanins (0.77–1.27 mg/g) from purple‐fleshed sweet potato juice, with comparable efficiency. Antioxidant activity correlated well with total phenolic content. All formulated ingredient matrices stabilized and preserved polyphenols for up to 24 weeks, even when stored at 37°C. Complexation with juice‐derived polyphenols did not significantly alter protein or carbohydrate profiles of the matrices. Sensory evaluation of the ingredient matrices suggested potential uses for a wide range of functional food products. DA - 2015/9// PY - 2015/9// DO - 10.1002/fsn3.234 VL - 3 IS - 5 SP - 415-424 SN - 2048-7177 KW - Anti-inflammatory KW - antioxidant KW - berries KW - polyphenols KW - stability KW - sweet potato ER - TY - JOUR TI - Metabolic footprinting of Lactobacillus buchneri strain LA1147 during anaerobic spoilage of fermented cucumbers AU - Johanningsmeier, Suzanne D. AU - McFeeters, Roger F. T2 - INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY AB - Lactobacillus buchneri has recently been associated with anaerobic spoilage of fermented cucumbers due to its ability to metabolize lactic acid into acetic acid and 1,2-propanediol. However, we have limited knowledge of other chemical components in fermented cucumber that may be related to spoilage and the unique metabolic capabilities of L. buchneri. Comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry metabolite profiling methods were applied for nontargeted detection of volatile and nonvolatile compounds to determine changes that occurred during anaerobic fermented cucumber spoilage by L. buchneri LA1147 and during reproduction of spoilage with natural microbiota. Univariate analysis of variance combined with hierarchial clustering analysis revealed 92 metabolites that changed during spoilage (P<0.01). Decreases were observed in mono and disaccharides, amino acids, nucleosides, long chain fatty acids, aldehydes, and ketones, and increases were observed in several alcohols and butanoic and pentanoic acids. Most of the metabolite changes preceded lactic acid utilization, indicating that lactic acid is not a preferred substrate for anaerobic spoilage organisms in fermented cucumbers. The ability to detect biochemical changes that preceded lactate utilization revealed citrulline, trehalose, and cellobiose as compounds that may signify metabolic activity of L. buchneri spoilage strains prior to any significant product degradation. DA - 2015/12/23/ PY - 2015/12/23/ DO - 10.1016/j.ijfoodmicro.2015.08.004 VL - 215 SP - 40-48 SN - 1879-3460 KW - GCxGC-ToFMS KW - Metabolomic platform KW - Lactic acid bacteria KW - Pickled vegetables KW - Spoilage organisms KW - Spoilage metabolites KW - L. buchneri metabolism ER - TY - JOUR TI - Expanding the biotechnology potential of lactobacilli through comparative genomics of 213 strains and associated genera AU - Sun, Zhihong AU - Harris, Hugh M. B. AU - McCann, Angela AU - Guo, Chenyi AU - Argimon, Silvia AU - Zhang, Wenyi AU - Yang, Xianwei AU - Jeffery, Ian B. AU - Cooney, Jakki C. AU - Kagawa, Todd F. AU - Liu, Wenjun AU - Song, Yuqin AU - Salvetti, Elisa AU - Wrobel, Agnieszka AU - Rasinkangas, Pia AU - Parkhill, Julian AU - Rea, Mary C. AU - O'Sullivan, Orla AU - Ritari, Jarmo AU - Douillard, Francois P. AU - Ross, R. Paul AU - Yang, Ruifu AU - Briner, Alexandra E. AU - Felis, Giovanna E. AU - Vos, Willem M. AU - Barrangou, Rodolphe AU - Klaenhammer, Todd R. AU - Caufield, Page W. AU - Cui, Yujun AU - Zhang, Heping AU - Paul W. O'Toole, T2 - NATURE COMMUNICATIONS AB - Lactobacilli are a diverse group of species that occupy diverse nutrient-rich niches associated with humans, animals, plants and food. They are used widely in biotechnology and food preservation, and are being explored as therapeutics. Exploiting lactobacilli has been complicated by metabolic diversity, unclear species identity and uncertain relationships between them and other commercially important lactic acid bacteria. The capacity for biotransformations catalysed by lactobacilli is an untapped biotechnology resource. Here we report the genome sequences of 213 Lactobacillus strains and associated genera, and their encoded genetic catalogue for modifying carbohydrates and proteins. In addition, we describe broad and diverse presence of novel CRISPR-Cas immune systems in lactobacilli that may be exploited for genome editing. We rationalize the phylogenomic distribution of host interaction factors and bacteriocins that affect their natural and industrial environments, and mechanisms to withstand stress during technological processes. We present a robust phylogenomic framework of existing species and for classifying new species. DA - 2015/9// PY - 2015/9// DO - 10.1038/ncomms9322 VL - 6 SP - SN - 2041-1723 ER - TY - JOUR TI - Effect of temperature and concentration on benzoyl peroxide bleaching efficacy and benzoic acid levels in whey protein concentrate AU - Smith, T. J. AU - Gerard, P. D. AU - Drake, M. A. T2 - JOURNAL OF DAIRY SCIENCE AB - Much of the fluid whey produced in the United States is a by-product of Cheddar cheese manufacture and must be bleached. Benzoyl peroxide (BP) is currently 1 of only 2 legal chemical bleaching agents for fluid whey in the United States, but benzoic acid is an unavoidable by-product of BP bleaching. Benzoyl peroxide is typically a powder, but new liquid BP dispersions are available. A greater understanding of the bleaching characteristics of BP is necessary. The objective of the study was to compare norbixin destruction, residual benzoic acid, and flavor differences between liquid whey and 80% whey protein concentrates (WPC80) bleached at different temperatures with 2 different benzoyl peroxides (soluble and insoluble). Two experiments were conducted in this study. For experiment 1, 3 factors (temperature, bleach type, bleach concentration) were evaluated for norbixin destruction using a response surface model–central composite design in liquid whey. For experiment 2, norbixin concentration, residual benzoic acid, and flavor differences were explored in WPC80 from whey bleached by the 2 commercially available BP (soluble and insoluble) at 5 mg/kg. In liquid whey, soluble BP bleached more norbixin than insoluble BP, especially at lower concentrations (5 and 10 mg/kg) at both cold (4°C) and hot (50°C) temperatures. The WPC80 from liquid whey bleached with BP at 50°C had lower norbixin concentration, benzoic acid levels, cardboard flavor, and aldehyde levels than WPC80 from liquid whey bleached with BP at 4°C. Regardless of temperature, soluble BP destroyed more norbixin at lower concentrations than insoluble BP. The WPC80 from soluble-BP-bleached wheys had lower cardboard flavor and lower aldehyde levels than WPC80 from insoluble-BP-bleached whey. This study suggests that new, soluble (liquid) BP can be used at lower concentrations than insoluble BP to achieve equivalent bleaching and that less residual benzoic acid remains in WPC80 powder from liquid whey bleached hot (50°C) than cold (4°C), which may provide opportunities to reduce benzoic acid residues in dried whey ingredients, expanding their marketability. DA - 2015/11// PY - 2015/11// DO - 10.3168/jds.2015-9890 VL - 98 IS - 11 SP - 7614-7627 SN - 1525-3198 KW - benzoyl peroxide KW - benzoic acid KW - whey protein concentrate KW - bleaching ER - TY - JOUR TI - An updated evolutionary classification of CRISPR-Cas systems AU - Makarova, K. S. AU - Wolf, Y. I. AU - Alkhnbashi, O. S. AU - Costa, F. AU - Shah, S. A. AU - Saunders, S. J. AU - Barrangou, R. AU - Brouns, S. J. J. AU - Charpentier, E. AU - Haft, D. H. AU - Horvath, P. AU - Moineau, S. AU - Mojica, F. J. M. AU - Terns, R. M. AU - Terns, M. P. AU - White, M. F. T2 - Nature Reviews. Microbiology DA - 2015/// PY - 2015/// VL - 13 IS - 11 SP - 722-736 ER - TY - JOUR TI - Acid Resistance and Molecular Characterization of Escherichia coli O157:H7 and Different Non-O157 Shiga Toxin-Producing E. coli Serogroups AU - Kim, Gwang-Hee AU - Breidt, Frederick AU - Fratamico, Pina AU - Oh, Deog-Hwan T2 - JOURNAL OF FOOD SCIENCE AB - The objective of this study was to compare the acid resistance (AR) of non-O157 Shiga toxin-producing Escherichia coli (STEC) strains belonging to serogroups O26, O45, O103, O104, O111, O121, and O145 with O157:H7 STEC isolated from various sources in 400 mM acetic acid solutions (AAS) at pH 3.2 and 30 °C for 25 min with or without glutamic acid. Furthermore, the molecular subgrouping of the STEC strains was analyzed with the repetitive sequence-based PCR (rep-PCR) method using a DiversiLab(TM) system. Results for a total of 52 strains ranged from 0.31 to 5.45 log reduction CFU/mL in the absence of glutamic acid and 0.02 to 0.33 CFU/mL in the presence of glutamic acid except for B447 (O26:H11), B452 (O45:H2), and B466 (O104:H4) strains. Strains belonging to serogroups O111, O121, and O103 showed higher AR than serotype O157:H7 strains in the absence of glutamic acid. All STEC O157:H7 strains exhibited a comparable DNA pattern with more than 95% similarity in the rep-PCR results, as did the strains belonging to serogroups O111 and O121. Surprisingly, the DNA pattern of B458 (O103:H2) was similar to that of O157:H7 strains with 82% similarity, and strain B458 strain showed the highest AR to AAS among the O103 strains with 0.44 log reduction CFU/mL without glutamic acid. In conclusion, STEC serotypes isolated from different sources exhibited diverse AR and genetic subtyping patterns. Results indicated that some non-O157 STEC strains may have higher AR than STEC O157:H7 strains under specific acidic conditions, and the addition of glutamic acid provided enhanced protection against exposure to AAS. DA - 2015/10// PY - 2015/10// DO - 10.1111/1750-3841.12996 VL - 80 IS - 10 SP - M2257-M2264 SN - 1750-3841 KW - acid resistance KW - E. coli non-O157 KW - DiversiLab (TM) system KW - glutamic acid KW - repetitive sequence-based PCR ER - TY - JOUR TI - The effect of microfiltration on color, flavor, and functionality of 80% whey protein concentrate AU - Qiu, Y. AU - Smith, T. J. AU - Foegeding, E. A. AU - Drake, M. A. T2 - Journal of Dairy Science AB - The residual annatto colorant in fluid Cheddar cheese whey is bleached to provide a neutral-colored final product. Currently, hydrogen peroxide (HP) and benzoyl peroxide are used for bleaching liquid whey. However, previous studies have shown that chemical bleaching causes off-flavor formation, mainly due to lipid oxidation and protein degradation. The objective of this study was to evaluate the efficacy of microfiltration (MF) on norbixin removal and to compare flavor and functionality of 80% whey protein concentrate (WPC80) from MF whey to WPC80 from whey bleached with HP or lactoperoxidase (LP). Cheddar cheese whey was manufactured from colored, pasteurized milk. The fluid whey was pasteurized and fat separated. Liquid whey was subjected to 4 different treatments: control (no bleaching; 50°C, 1 h), HP (250 mg of HP/kg; 50°C, 1 h), and LP (20 mg of HP/kg; 50°C, 1 h), or MF (microfiltration; 50°C, 1 h). The treated whey was then ultrafiltered, diafiltered, and spray-dried to 80% concentrate. The entire experiment was replicated 3 times. Proximate analyses, color, functionality, descriptive sensory and instrumental volatile analysis were conducted on WPC80. The MF and HP- and LP-bleached WPC80 displayed a 39.5, 40.9, and 92.8% norbixin decrease, respectively. The HP and LP WPC80 had higher cardboard flavors and distinct cabbage flavor compared with the unbleached and MF WPC80. Volatile compound results were consistent with sensory results. The HP and LP WPC80 were higher in lipid oxidation compounds (especially heptanal, hexanal, pentanal, 1-hexen-3-one, 2-pentylfuran, and octanal) compared with unbleached and MF WPC80. All WPC80 had >85% solubility across the pH range of 3 to 7. The microstructure of MF gels determined by confocal laser scanning showed an increased protein particle size in the gel network. MF WPC80 also had larger storage modulus values, indicating higher gel firmness. Based on bleaching efficacy comparable to chemical bleaching with HP, flavor, and functionality results, MF is a viable alternative to chemical or enzymatic bleaching of fluid whey. DA - 2015/// PY - 2015/// DO - 10.3168/jds.2014-9174 VL - 98 IS - 9 SP - 5862-5873 ER - TY - JOUR TI - RNA-Seq analysis and annotation of a draft blueberry genome assembly identifies candidate genes involved in fruit ripening, biosynthesis of bioactive compounds, and stage-specific alternative splicing AU - Gupta, V. AU - Estrada, A. D. AU - Blakley, I. AU - Reid, R. AU - Patel, K. AU - Meyer, M. D. AU - Andersen, S. U. AU - Brown, A. F. AU - Lila, M. A. AU - Loraine, A. E. T2 - Gigascience DA - 2015/// PY - 2015/// VL - 4 ER - TY - JOUR TI - Mitochondrial DNA Fragmentation as a Molecular Tool to Monitor Thermal Processing of Plant-Derived, Low-Acid Foods, and Biomaterials AU - Caldwell, Jane M. AU - Perez-Diaz, Ilenys M. AU - Sandeep, K. P. AU - Simunovic, Josip AU - Harris, Keith AU - Osborne, Jason A. AU - Hassan, Hosni M. T2 - JOURNAL OF FOOD SCIENCE AB - Cycle threshold (Ct) increase, quantifying plant-derived DNA fragmentation, was evaluated for its utility as a time-temperature integrator. This novel approach to monitoring thermal processing of fresh, plant-based foods represents a paradigm shift. Instead of using quantitative polymerase chain reaction (qPCR) to detect pathogens, identify adulterants, or authenticate ingredients, this rapid technique was used to quantify the fragmentation of an intrinsic plant mitochondrial DNA (mtDNA) gene over time-temperature treatments. Universal primers were developed which amplified a mitochondrial gene common to plants (atp1). These consensus primers produced a robust qPCR signal in 10 vegetables, 6 fruits, 3 types of nuts, and a biofuel precursor. Using sweet potato (Ipomoea batatas) puree as a model low-acid product and simple linear regression, Ct value was highly correlated to time-temperature treatment (R(2) = 0.87); the logarithmic reduction (log CFU/mL) of the spore-forming Clostridium botulinum surrogate, Geobacillus stearothermophilus (R(2) = 0.87); and cumulative F-value (min) in a canned retort process (R(2) = 0.88), all comparisons conducted at 121 °C. D121 and z-values were determined for G. stearothermophilus ATCC 7953 and were 2.71 min and 11.0 °C, respectively. D121 and z-values for a 174-bp universal plant amplicon were 11.3 min and 9.17 °C, respectively, for mtDNA from sweet potato puree. We present these data as proof-of-concept for a molecular tool that can be used as a rapid, presumptive method for monitoring thermal processing in low-acid plant products. DA - 2015/8// PY - 2015/8// DO - 10.1111/1750-3841.12937 VL - 80 IS - 8 SP - M1804-M1814 SN - 1750-3841 KW - mitochondrial DNA KW - quantitative PCR KW - thermal processing ER - TY - JOUR TI - Aerosolization of a Human Norovirus Surrogate, Bacteriophage MS2, during Simulated Vomiting AU - Tung-Thompson, Grace AU - Libera, Dominic A. AU - Koch, Kenneth L. AU - Reyes, Francis L., III AU - Jaykus, Lee-Ann T2 - PLOS ONE AB - Human noroviruses (NoV) are the leading cause of acute gastroenteritis worldwide. Epidemiological studies of outbreaks have suggested that vomiting facilitates transmission of human NoV, but there have been no laboratory-based studies characterizing the degree of NoV release during a vomiting event. The purpose of this work was to demonstrate that virus aerosolization occurs in a simulated vomiting event, and to estimate the amount of virus that is released in those aerosols. A simulated vomiting device was constructed at one-quarter scale of the human body following similitude principles. Simulated vomitus matrices at low (6.24 mPa*s) and high (177.5 mPa*s) viscosities were inoculated with low (108 PFU/mL) and high (1010 PFU/mL) concentrations of bacteriophage MS2 and placed in the artificial “stomach” of the device, which was then subjected to scaled physiologically relevant pressures associated with vomiting. Bio aerosols were captured using an SKC Biosampler. In low viscosity artificial vomitus, there were notable differences between recovered aerosolized MS2 as a function of pressure (i.e., greater aerosolization with increased pressure), although this was not always statistically significant. This relationship disappeared when using high viscosity simulated vomitus. The amount of MS2 aerosolized as a percent of total virus “vomited” ranged from 7.2 x 10-5 to 2.67 x 10-2 (which corresponded to a range of 36 to 13,350 PFU total). To our knowledge, this is the first study to document and measure aerosolization of a NoV surrogate in a similitude-based physical model. This has implications for better understanding the transmission dynamics of human NoV and for risk modeling purposes, both of which can help in designing effective infection control measures. DA - 2015/8/19/ PY - 2015/8/19/ DO - 10.1371/journal.pone.0134277 VL - 10 IS - 8 SP - SN - 1932-6203 ER - TY - JOUR TI - US products-dry-cured hams AU - Hanson, Dana AU - Rentfrow, G. AU - Schilling, M. W. AU - Mikel, W. B. AU - Stalder, K. J. AU - Berry, N. L. T2 - Handbook of fermented meat and poultry, 2nd edition AB - Country ham production in the United States began as early as the 1500s. European explorers and settlers brought pigs to North America to provide a source of food. Without refrigeration, it was critical to preserve the meat. The United States Department of Agriculture Food Safety and Inspection Service (USDA-FSIS) defines country ham as an uncooked, cured, dried, smoked, or unsmoked meat product made from a single piece of pork ham muscle. The dry-cure mix (salt, sugar, sodium nitrate and/or sodium nitrite) is applied to the surface of the raw meat in a sufficient quantity to ensure the finished dry product has an internal salt content >4%. Most country ham has a finished salt percentage of 5–8%. The salted raw hams are then maintained (cured) at temperatures of 1–4°C for approximately 40–50 days. After this cure step, the hams are maintained at 10–12°C for an additional 14–21 days. The final production step is to dry or age the ham at 30–35°C. This step can be as short as 20 days or as long as an additional year. Country ham flavor develops and intensifies over time, so the aging time is dependent on the desired flavor profile. Finished country ham will have a water activity of 0.85–0.91. The lowered water activity and salt content ensure the safety and shelf stability of the finished product. In the United States, most consumers of country ham will cook it prior to eating. Common methods of cooking include baking the whole ham and pan frying thin slices of meat. However, a small percentage of long-aged and low-water-activity country ham is consumed uncooked, similar to European dry-cured ham products. DA - 2015/// PY - 2015/// DO - 10.1002/9781118522653.ch40 SP - 347–354 ER - TY - JOUR TI - JFS Special Issue: 75 years of advancing food science, and preparing for the next 75 CRISPR-based technologies and the future of food science AU - Selle, K. AU - Barrangou, R. T2 - Journal of Food Science DA - 2015/// PY - 2015/// VL - 80 IS - 11 SP - R2367-2372 ER - TY - JOUR TI - Hepatoprotective Activity of Easter Lily (Lilium longiflorum Thunb.) Bulb Extracts AU - Tang, Wenping AU - Munafo, John P., Jr. AU - Palatini, Kimberly AU - Esposito, Debora AU - Huang, Mou-Tuan AU - Komarnytsky, Slavko AU - Ho, Chi-Tang AU - Gianfagna, Thomas J. T2 - JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY AB - The hepatoprotective activities of two different extracts, a hydroethanolic crude bulb extract (CB) and a steroidal glycoside-rich 1-butanol extract (BuOH), prepared from the bulbs of Easter lily (Lilium longiflorum Thunb.), were evaluated in a 24 week study in the female KK.Cg-A(y)/J Type 2 diabetic mouse model. Animals were divided into six groups (n = 16): control mice received Easter lily bulb extract-free drinking water together with a low- or high-fat diet (diabetic control); drinking water for the remaining groups was supplemented with CB extract (1%), BuOH extract (0.1 or 0.2%), and reference drug Metformin (0.001%), together with a high-fat diet. Both CB and BuOH extract treatment groups exhibited significantly improved liver function based on comparisons of triglycerides [diabetic 219 ± 34 mg/dL, CB 131 ± 27 mg/dL, BuOH(0.2%) 114 ± 35 mg/dL], CB total cholesterol (TC) (diabetic 196 ± 12 mg/dL, CB 159 ± 5 mg/dL), average liver mass [diabetic 2.96 ± 0.13 g, CB 2.58 ± 0.08 g, BuOH(0.1%) 2.48 ± 0.13 g], alanine transferase [diabetic 74 ± 5 units/L, CB 25 ± 1 units/L, BuOH(0.1%) 45 ± 1 units/L], and histological examinations. Glucose metabolism was improved only in CB, which was confirmed by oral glucose tolerance tests (OGTT) in diet-induced obese C57BL/6J mice exposed to CB extract. These data suggest that steroidal glycosides 1-5 might play a role in the hepatoprotective activity of the BuOH extracts, while the results of the TC measurements and OGTT study indicate that other constituents present in the CB extract are responsible for its hypocholesterolemic and hypoglycemic activity. DA - 2015/11/11/ PY - 2015/11/11/ DO - 10.1021/acs.jafc.5b04078 VL - 63 IS - 44 SP - 9722-9728 SN - 1520-5118 KW - Lilium longiflorum Thunb KW - Easter lily KW - steroidal glycoside KW - hepatoprotective activity KW - hypoglycemic activity ER - TY - JOUR TI - Transforming Structural Breakdown into Sensory Perception of Texture AU - Foegeding, E. Allen AU - Vinyard, Christopher J. AU - Essick, Gregory AU - Guest, Steve AU - Campbell, Caroline T2 - JOURNAL OF TEXTURE STUDIES AB - Abstract Food structure research has historically focused on building and stabilizing specific structural elements. Precise execution of unit operations in food processing is required to ensure consistent structure formation, but does not explain why a particular structure is desirable. The pattern by which food structure breaks down during oral processing, generating texture, flavor and taste, determines the acceptability of a food. However, breakdown patterns and textural perception associated with desirable food qualities are not well understood. Food materials are classified based on their physical state (liquids, semisolids or solids) and interactions with surfaces (adhesive or nonadhesive). Physical and chemical processes during oral processing are used to further characterize each food category. Typical processes include particle reduction, dissolution, hydration, removal of food from oral surface and bolus formation. This review presents a framework for classifying food materials and material transformations during oral processing in order to identify their contributions to texture perception. Practical Applications There is a constant desire to reformulate or modify foods based on economic, sustainability, health and dietary considerations. One goal of a reformulated food is to change the composition but cause no significant changes in flavor, taste and textural properties associated with desirability. An alternative goal is to modulate a specific textural element to make the food more acceptable to a target population (e.g., those with compromised chewing ability). Both goals require an understanding of the fundamental elements that cause specific textural properties. This manuscript presents an overview of how the breakdown of food structure during oral processing generates the cognition of textural properties. Once textural properties are understood based on specific elements of structural transitions, this will allow for development of textures for specific market segments (e.g., vegan, gluten free or swallowing disorders) as well as the ability to control nutrient bioavailability and flavor release. DA - 2015/6// PY - 2015/6// DO - 10.1111/jtxs.12105 VL - 46 IS - 3 SP - 152-170 SN - 1745-4603 KW - Food structure KW - mechanoreceptors KW - oral processing KW - sensory analysis KW - texture ER - TY - JOUR TI - The Development and Validation of the Childhood Obesity Prevention Self-Efficacy (COP-SE) Survey AU - Cooke, Natalie K. AU - Nietfeld, John L. AU - Goodell, L. Suzanne T2 - CHILDHOOD OBESITY AB - Background: Physicians can play an important role in preventing and treating childhood obesity. There are currently no validated measures of medical students' self-efficacy in these skills; therefore, we sought to develop a valid and reliable computerized survey to measure medical students' self-efficacy in skills needed to prevent and treat childhood obesity. Methods: We developed the Childhood Obesity Prevention Self-Efficacy (COP-SE) survey with input from two expert panels and cognitive interviews with medical students. We administered the 43-item COP-SE computerized survey to a nation-wide sample of medical students. Results: The final sample consisted of 444 medical students from 53 medical schools. Exploratory factor analysis revealed a two-factor structure with a correlation of 0.637 between factors and high reliability within factors. The correlation between the COP-SE and a measure of general self-efficacy was moderate (0.648), and reliability within factors was high (Factor 1=0.946; Factor 2=0.927). Conclusions: The 18-item COP-SE is a valid and reliable measure of childhood obesity prevention self-efficacy. Factor 1 assesses self-efficacy in nutrition counseling, and Factor 2 measures self-efficacy to assess readiness to change and initiate nutrition lifestyle changes. The correlation between the COP-SE and a measure of general self-efficacy indicates that the COP-SE is a distinct, valid assessment of domain-specific self-efficacy. The high reliability of items within factors indicates the items measure the same constructs. Therefore, medical schools can use this valid and reliable instrument as a formative or summative assessment of students' self-efficacy in childhood obesity prevention and treatment. DA - 2015/4/1/ PY - 2015/4/1/ DO - 10.1089/chi.2014.0103 VL - 11 IS - 2 SP - 114-121 SN - 2153-2176 ER - TY - JOUR TI - Sortase-deficient lactobacilli: effect on immunomodulation and gut retention AU - Call, Emma K. AU - Goh, Yong Jun AU - Selle, Kurt AU - Klaenhammer, Todd R. AU - O'Flaherty, Sarah T2 - MICROBIOLOGY-SGM AB - Surface proteins of probiotic microbes, including Lactobacillus acidophilus and Lactobacillus gasseri, are believed to promote retention in the gut and mediate host–bacterial communications. Sortase, an enzyme that covalently couples a subset of extracellular proteins containing an LPXTG motif to the cell surface, is of particular interest in characterizing bacterial adherence and communication with the mucosal immune system. A sortase gene, srtA, was identified in L. acidophilus NCFM (LBA1244) and L. gasseri ATCC 33323 (LGAS_0825). Additionally, eight and six intact sortase-dependent proteins were predicted in L. acidophilus and L. gasseri, respectively. Due to the role of sortase in coupling these proteins to the cell wall, ΔsrtA deletion mutants of L. acidophilus and L. gasseri were created using the upp-based counterselective gene replacement system. Inactivation of sortase did not cause significant alteration in growth or survival in simulated gastrointestinal juices. Meanwhile, both ΔsrtA mutants showed decreased adhesion to porcine mucin in vitro. Murine dendritic cells exposed to the ΔsrtA mutant of L. acidophilus or L. gasseri induced lower levels of pro-inflammatory cytokines TNF-α and IL-12, respectively, compared with the parent strains. In vivo co-colonization of the L. acidophilus ΔsrtA mutant and its parent strain in germ-free 129S6/SvEv mice resulted in a significant one-log reduction of the ΔsrtA mutant population. Additionally, a similar reduction of the ΔsrtA mutant was observed in the caecum. This study shows for the first time that sortase-dependent proteins contribute to gut retention of probiotic microbes in the gastrointestinal tract. DA - 2015/2// PY - 2015/2// DO - 10.1099/mic.0.000007 VL - 161 SP - 311-321 SN - 1350-0872 ER - TY - JOUR TI - Sensory Perception, Nutritional Role, and Challenges of Flavored Milk for Children and Adults AU - Li, Xiaomeng E. AU - Drake, MaryAnne T2 - JOURNAL OF FOOD SCIENCE AB - Milk and milk products provide essential nutrients for both adults and children. However, overall milk consumption of both adults and children does not meet the recommendations from Dietary Guidelines for Americans. Flavored milk can increase milk consumption for children and adolescents, but the added sugar content raises concern. Since the removal or reduction of flavored milk decreases milk consumption for children, it is important to understand all aspects of flavored milk in order to increase milk consumption while minimizing sugar intake. This review will address adult and children perception of flavored milk as well as its nutrition, regulations in school meal programs, and challenges. Understanding the sensory perception of both adults and children for flavored milk can help food developers and manufacturers to address attractive attributes while reducing the sugar content to meet the needs of a healthy diet. DA - 2015/4// PY - 2015/4// DO - 10.1111/1750-3841.12828 VL - 80 IS - 4 SP - R665-R670 SN - 1750-3841 KW - children KW - flavored milk KW - milk consumption KW - sugar content KW - sweeteners ER - TY - JOUR TI - Registration of 'Sugg' Peanut AU - Isleib, Thomas G. AU - Milla-Lewis, Susana R. AU - Pattee, Harold E. AU - Copeland, Susan C. AU - Zuleta, M. Carolina AU - Shew, Barbara B. AU - Hollowell, Joyce E. AU - Sanders, Timothy H. AU - Dean, Lisa O. AU - Hendrix, Keith W. AU - Balota, Maria AU - Chapin, Jay W. AU - Monfort, W. Scott T2 - JOURNAL OF PLANT REGISTRATIONS AB - ‘Sugg’ (Reg. No. CV-125, PI 666112) is a large-seeded virginia-type peanut (Arachis hypogaea L. subsp. hypogaea var. hypogaea) cultivar with partial resistance to four diseases that occur commonly in the Virginia–Carolina production area: early leafspot caused by Cercospora arachidicola S. Hori, Cylindrocladium black rot caused by Cylindrocladium parasiticum Crous, Wingfield & Alfenas, Sclerotinia blight caused by Sclerotinia minor Jagger, and tomato spotted wilt caused by the Tomato spotted wilt tospovirus. Sugg was developed as part of a program of selection for multiple disease resistance funded by growers, seed dealers, shellers, and processors. Sugg was tested under the experimental designation N03091T and released by the North Carolina Agricultural Research Service (NCARS) in 2009. Sugg was tested by the NCARS, the Virginia Agricultural Experiment Station, and five other state agricultural experiment stations and the USDA–ARS units participating in the Uniform Peanut Performance Tests. Sugg has alternate branching pattern, intermediate runner growth habit, medium green foliage, and high contents of fancy pods and medium virginia-type seeds. It has seeds with pink testa averaging 957 mg seed−1, approximately 40% jumbo and 46% fancy pods, and extra-large kernel content of ∼47%. Sugg is named in honor of Norfleet “Fleet” Sugg and the late Joseph “Joe” Sugg, cousins who served consecutively as executive directors of the North Carolina Peanut Growers Association from 1966 through 1993. DA - 2015/1// PY - 2015/1// DO - 10.3198/jpr2013.09.0059crc VL - 9 IS - 1 SP - 44-52 SN - 1940-3496 ER - TY - JOUR TI - Parents' and Children's Acceptance of Skim Chocolate Milks Sweetened by Monk Fruit and Stevia Leaf Extracts AU - Li, X. E. AU - Lopetcharat, K. AU - Drake, M. A. T2 - JOURNAL OF FOOD SCIENCE AB - Abstract Chocolate milk increases milk consumption of children, but high sugar content raises health concerns. Interest in sugar reduction and parents’ preference for natural sweeteners necessitates further research on natural nonnutritive sweeteners. However, it is important to maintain consumer acceptability, especially for children, while reducing sugar in chocolate milk. The objectives of this study were to identify the sweetness intensity perception of stevia leaf (STV) and monk fruit (MK) extracts in skim chocolate milk (SCM), to evaluate STV and MK as the sole or partial sweetener source for SCM for young adults (19 to 35 y) and children (5 to 13 y), and to determine if information on natural nonnutritive sweeteners impacted parents’ acceptability of SCM. Power function and 2‐alternative forced choice studies were used to determine the iso‐sweetness of nonnutritive sweeteners to a sucrose control in SCM (51.4 g/L, SUC control). Young adults ( n = 131) evaluated 9 different SCM (SUC control, STV, MK, STV:sucrose blends, or MK:sucrose blends) in a completely randomized 2‐d test. Children ( n = 167) evaluated SUC control SCM and SCM with 39.7 g/L sucrose and 46 mg/L MK (MK25) or 30 mg/L STV (STV25). Parents evaluated SUC control, MK25, and STV25 in a balanced crossover design with a 40‐d wait time between primed or unprimed ballots. Chocolate milks solely sweetened by nonnutritive sweeteners were less acceptable compared with SUC control by young adults. MK25 and STV25 were acceptable by young adults and children. The presentation of chocolate milk label information had different effects on parental acceptance. Traditional parents preferred sucrose sweetened SCM, and label conscious parents preferred SCM with natural nonnutritive sweeteners. DA - 2015/5// PY - 2015/5// DO - 10.1111/1750-3841.12835 VL - 80 IS - 5 SP - S1083-S1092 SN - 1750-3841 KW - chocolate milk KW - natural nonnutritive sweetener KW - priming KW - power function KW - sugar reduction ER - TY - JOUR TI - Microbial Ecology of Watery Kimchi AU - Kyung, Kyu Hang AU - Medina Pradas, Eduardo AU - Kim, Song Gun AU - Lee, Yong Jae AU - Kim, Kyong Ho AU - Choi, Jin Joo AU - Cho, Joo Hyong AU - Chung, Chang Ho AU - Barrangou, Rodolphe AU - Breidt, Frederick T2 - JOURNAL OF FOOD SCIENCE AB - Abstract The biochemistry and microbial ecology of 2 similar types of watery ( mul ) kimchi, containing sliced and unsliced radish and vegetables ( nabak and dongchimi , respectively), were investigated. Samples from kimchi were fermented at 4, 10, and 20 °C were analyzed by plating on differential and selective media, high‐performance liquid chromatography, and high‐throughput DNA sequencing of 16S rDNA. Nabak kimchi showed similar trends as dongchimi , with increasing lactic and acetic acids and decreasing pH for each temperature, but differences in microbiota were apparent. Interestingly, bacteria from the Proteobacterium phylum, including Enterobacteriaceae , decreased more rapidly during fermentation at 4 °C in nabak cabbage fermentations compared with dongchimi . Although changes for Proteobacterium and Enterobacteriaceae populations were similar during fermentation at 10 and 20 °C, the homolactic stage of fermentation did not develop for the 4 and 10 °C samples of both nabak and dongchimi during the experiment. These data show the differences in biochemistry and microbial ecology that can result from preparation method and fermentation conditions of the kimchi, which may impact safety ( Enterobacteriaceae populations may include pathogenic bacteria) and quality (homolactic fermentation can be undesirable, if too much acid is produced) of the product. In addition, the data also illustrate the need for improved methods for identifying and differentiating closely related lactic acid bacteria species using high‐throughput sequencing methods. DA - 2015/5// PY - 2015/5// DO - 10.1111/1750-3841.12848 VL - 80 IS - 5 SP - M1031-M1038 SN - 1750-3841 KW - high-throughput sequencing KW - microbial ecology KW - watery kimchi ER - TY - JOUR TI - Human Norovirus as a Foodborne Pathogen: Challenges and Developments AU - Moore, Matthew D. AU - Goulter, Rebecca M. AU - Jaykus, Lee-Ann T2 - ANNUAL REVIEW OF FOOD SCIENCE AND TECHNOLOGY, VOL 6 AB - Human noroviruses (NoVs) are the leading cause of foodborne illness in the United States, and they exact a considerable human and economic burden worldwide. In fact, the many challenging aspects of human NoV have caused some to call it the nearly perfect foodborne pathogen. In this review, a brief overview of NoVs and their genetic structure is provided. Additionally, the challenges and recent developments related to human NoVs regarding viral evolution, transmission, epidemiology, outbreak identification, cultivation, animal and human models, and detection are presented. DA - 2015/// PY - 2015/// DO - 10.1146/annurev-food-022814-015643 VL - 6 SP - 411-433 SN - 1941-1413 KW - epidemiology KW - detection KW - evolution KW - cell culture KW - infectivity ER - TY - JOUR TI - Harnessing CRISPR-Cas systems for bacterial genome editing AU - Selle, Kurt AU - Barrangou, Rodolphe T2 - TRENDS IN MICROBIOLOGY AB - Manipulation of genomic sequences facilitates the identification and characterization of key genetic determinants in the investigation of biological processes. Genome editing via clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) constitutes a next-generation method for programmable and high-throughput functional genomics. CRISPR-Cas systems are readily reprogrammed to induce sequence-specific DNA breaks at target loci, resulting in fixed mutations via host-dependent DNA repair mechanisms. Although bacterial genome editing is a relatively unexplored and underrepresented application of CRISPR-Cas systems, recent studies provide valuable insights for the widespread future implementation of this technology. This review summarizes recent progress in bacterial genome editing and identifies fundamental genetic and phenotypic outcomes of CRISPR targeting in bacteria, in the context of tool development, genome homeostasis, and DNA repair. DA - 2015/4// PY - 2015/4// DO - 10.1016/j.tim.2015.01.008 VL - 23 IS - 4 SP - 225-232 SN - 1878-4380 KW - genome editing KW - CRISPR-Cas KW - Cas9 ER - TY - JOUR TI - Genetic Mechanisms of Prebiotic Oligosaccharide Metabolism in Probiotic Microbes AU - Goh, Yong Jun AU - Klaenhammer, Todd R. T2 - ANNUAL REVIEW OF FOOD SCIENCE AND TECHNOLOGY, VOL 6 AB - Recent insights into the relationship between the human gut and its resident microbiota have revolutionized our appreciation of this symbiosis and its impact on health and disease development. Accumulating evidence on probiotic and prebiotic interventions has demonstrated promising effects on promoting gastrointestinal health by modulating the microbiota toward the enrichment of beneficial microorganisms. However, the precise mechanisms of how prebiotic nondigestible oligosaccharides are metabolized by these beneficial microbes in vivo remain largely unknown. Genome sequencing of probiotic lactobacilli and bifidobacteria has revealed versatile carbohydrate metabolic gene repertoires dedicated to the catabolism of various oligosaccharides. In this review, we highlight recent findings on the genetic mechanisms involved in the utilization of prebiotic fructooligosaccharides, β-galactooligosaccharides, human milk oligosaccharides, and other prebiotic candidates by these probiotic microbes. DA - 2015/// PY - 2015/// DO - 10.1146/annurev-food-022814-015706 VL - 6 SP - 137-156 SN - 1941-1421 KW - Lactobacillus KW - Bifidobacterium KW - fructooligosaccharide KW - galactooligosaccharide KW - prebiotic KW - probiotic ER - TY - JOUR TI - Examining Extrinsic Factors that Influence Product Acceptance: A Review AU - Li, X. E. AU - Jervis, S. M. AU - Drake, M. A. T2 - JOURNAL OF FOOD SCIENCE AB - Drivers of liking (DOL) studies are useful for product development to formulate acceptable products; however, DOL alone are insufficient for understanding why a product is purchased and repurchased, which is ultimately the indication of a successful product. Ultimately sensory attributes drive product success (that is, repeat and continued purchase). However, ignoring the importance of extrinsic factors may neglect the vital product attributes responsible for the initial purchase, which may in turn, affect repeat purchase. The perception of sensory attributes assessed by DOL is mitigated by external perceptions of quality. If the sensory attributes do not deliver based upon the quality cues, the product will not be acceptable. Four key extrinsic factors that affect DOL are the perceived satiety, brand and labeling, price, and the emotional impact to decision making. In order to more thoroughly understand what the DOL for a product is, these 4 product cues should be considered in conjunction with sensory attribute perception to gain a holistic understanding of product acceptance. DA - 2015/5// PY - 2015/5// DO - 10.1111/1750-3841.12852 VL - 80 IS - 5 SP - R901-R909 SN - 1750-3841 KW - brand KW - decision making KW - drivers of liking KW - emotions KW - perceived satiety KW - willingness-to-pay ER - TY - JOUR TI - Decolorization of Cheddar cheese whey by activated carbon AU - Zhang, Yue AU - Campbell, Rachel AU - Drake, Mary Anne AU - Zhong, Qixin T2 - JOURNAL OF DAIRY SCIENCE AB - Colored Cheddar whey is a source for whey protein recovery and is decolorized conventionally by bleaching, which affects whey protein quality. Two activated carbons were studied in the present work as physical means of removing annatto (norbixin) in Cheddar cheese whey. The color and residual norbixin content of Cheddar whey were reduced by a higher level of activated carbon at a higher temperature between 25 and 55°C and a longer time. Activated carbon applied at 40 g/L for 2 h at 30°C was more effective than bleaching by 500 mg/L of hydrogen peroxide at 68°C. The lowered temperature in activated-carbon treatments had less effect on protein structure as investigated for fluorescence spectroscopy and volatile compounds, particularly oxidation products, based on gas chromatography-mass spectrometry. Activated carbon was also reusable, removing more than 50% norbixin even after 10 times of regeneration, which showed great potential for decolorizing cheese whey. DA - 2015/5// PY - 2015/5// DO - 10.3168/jds.2014-9159 VL - 98 IS - 5 SP - 2982-2991 SN - 1525-3198 KW - activated carbon KW - norbixin (annatto) removal KW - Cheddar cheese whey KW - bleaching KW - volatile analysis ER - TY - JOUR TI - Charge related astringency of chitosans AU - Luck, Paige AU - Varum, Kjell M. AU - Foegeding, E. Allen T2 - FOOD HYDROCOLLOIDS AB - Chitosan is a biopolymer that can be used in food applications such as forming edible coatings and nanoparticles. It is a linear polysaccharide composed of neutral (acetylated) and positively (deacetylated) sugar units and can be prepared with varying content of the two units. One factor limiting the use of chitosan is causing an unacceptable level of astringency. We hypothesized that astringency was related to the positive charge density and that decreasing the charge density would decrease astringency. Chitosans with fraction of acetylated units (FA) ranging from 0.01 to 0.49 (percentage degree of acetylation from 1 to 49) were analyzed for astringency using a time intensity sensory method. In addition, the ability of chitosans to precipitate saliva proteins was determined. Increasing FA from 0.01 to 0.49 caused a decrease in maximum astringency sensation from 10.9 to 7.2. Mucins and other saliva proteins were precipitated by chitosans but a simple association with astringency intensity was not observed. It can be concluded that altering the charge density of chitosans is a means to alter chitosan astringency. DA - 2015/6// PY - 2015/6// DO - 10.1016/j.foodhyd.2015.02.024 VL - 48 SP - 174-178 SN - 1873-7137 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-84924240333&partnerID=MN8TOARS KW - Astringency KW - Chitosan KW - Saliva KW - Proline-rich proteins KW - Mucins ER - TY - JOUR TI - Characterization of the microbial diversity in yacon spontaneous fermentation at 20 degrees C AU - Reina, L. D. AU - Perez-Diaz, I. M. AU - Breidt, F. AU - Azcarate-Peril, M. A. AU - Medina, E. AU - Butz, N. T2 - INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY AB - The prebiotic fructooligosaccharide content of yacon makes this root an attractive alternative for the supplementation of a variety of food products. The preservation of yacon by fermentation has been proposed as an alternative to increase the probiotic content of the root concomitantly with its shelf life. Thus the fermented yacon could have significant functional content. The objective of this research was to characterize the biochemistry and microbiology of spontaneous yacon fermentation with 2% NaCl and define the viability of the proposed process. The biochemical analysis of spontaneous heterolactic fermentation of yacon showed a progressive drop in pH with increased lactic and acetic acids, and the production of mannitol during fermentation. The microbial ecology of yacon fermentation was investigated using culture-dependent and culture-independent methods. Bacterial cell counts revealed a dominance of lactic acid bacteria (LAB) over yeasts, which were also present during the first 2 days of the fermentation. Results showed that the heterofermentative LAB were primarily Leuconostoc species, thus it presents a viable method to achieve long term preservation of this root. DA - 2015/6/16/ PY - 2015/6/16/ DO - 10.1016/j.ijfoodmicro.2015.03.007 VL - 203 SP - 35-40 SN - 1879-3460 KW - Fermented vegetables KW - Lactic acid bacteria KW - Heterolactic fermentation ER - TY - JOUR TI - Characterization and evolution of Salmonella CRISPR-Cas systems AU - Shariat, Nikki AU - Timme, Ruth E. AU - Pettengill, James B. AU - Barrangou, Rodolphe AU - Dudley, Edward G. T2 - MICROBIOLOGY-SGM AB - Prokaryotic CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated genes) systems provide adaptive immunity from invasive genetic elements and encompass three essential features: (i) cas genes, (ii) a CRISPR array composed of spacers and direct repeats and (iii) an AT-rich leader sequence upstream of the array. We performed in-depth sequence analysis of the CRISPR-Cas systems in >600 Salmonella, representing four clinically prevalent serovars. Each CRISPR-Cas feature is extremely conserved in the Salmonella, and the CRISPR1 locus is more highly conserved than CRISPR2. Array composition is serovar-specific, although no convincing evidence of recent spacer acquisition against exogenous nucleic acids exists. Only 12 % of spacers match phage and plasmid sequences and self-targeting spacers are associated with direct repeat variants. High nucleotide identity (>99.9 %) exists across the cas operon among isolates of a single serovar and in some cases this conservation extends across divergent serovars. These observations reflect historical CRISPR-Cas immune activity, showing that this locus has ceased undergoing adaptive events. Intriguingly, the high level of conservation across divergent serovars shows that the genetic integrity of these inactive loci is maintained over time, contrasting with the canonical view that inactive CRISPR loci degenerate over time. This thorough characterization of Salmonella CRISPR-Cas systems presents new insights into Salmonella CRISPR evolution, particularly with respect to cas gene conservation, leader sequences, organization of direct repeats and protospacer matches. Collectively, our data suggest that Salmonella CRISPR-Cas systems are no longer immunogenic; rather, their impressive conservation indicates they may have an alternative function in Salmonella. DA - 2015/2// PY - 2015/2// DO - 10.1099/mic.0.000005 VL - 161 SP - 374-386 SN - 1465-2080 ER - TY - JOUR TI - CRISPR-based screening of genomic island excision events in bacteria AU - Selle, Kurt AU - Klaenhammer, Todd R. AU - Barrangou, Rodolphe T2 - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA AB - Genomic analysis of Streptococcus thermophilus revealed that mobile genetic elements (MGEs) likely contributed to gene acquisition and loss during evolutionary adaptation to milk. Clustered regularly interspaced short palindromic repeats-CRISPR-associated genes (CRISPR-Cas), the adaptive immune system in bacteria, limits genetic diversity by targeting MGEs including bacteriophages, transposons, and plasmids. CRISPR-Cas systems are widespread in streptococci, suggesting that the interplay between CRISPR-Cas systems and MGEs is one of the driving forces governing genome homeostasis in this genus. To investigate the genetic outcomes resulting from CRISPR-Cas targeting of integrated MGEs, in silico prediction revealed four genomic islands without essential genes in lengths from 8 to 102 kbp, totaling 7% of the genome. In this study, the endogenous CRISPR3 type II system was programmed to target the four islands independently through plasmid-based expression of engineered CRISPR arrays. Targeting lacZ within the largest 102-kbp genomic island was lethal to wild-type cells and resulted in a reduction of up to 2.5-log in the surviving population. Genotyping of Lac(-) survivors revealed variable deletion events between the flanking insertion-sequence elements, all resulting in elimination of the Lac-encoding island. Chimeric insertion sequence footprints were observed at the deletion junctions after targeting all of the four genomic islands, suggesting a common mechanism of deletion via recombination between flanking insertion sequences. These results established that self-targeting CRISPR-Cas systems may direct significant evolution of bacterial genomes on a population level, influencing genome homeostasis and remodeling. DA - 2015/6/30/ PY - 2015/6/30/ DO - 10.1073/pnas.1508525112 VL - 112 IS - 26 SP - 8076-8081 SN - 0027-8424 KW - CRISPR KW - lactic acid bacteria KW - transposons KW - IS-elements KW - Cas9 ER - TY - JOUR TI - CRISPR Immunity Drives Rapid Phage Genome Evolution in Streptococcus thermophilus AU - Paez-Espino, David AU - Sharon, Itai AU - Morovic, Wesley AU - Stahl, Buffy AU - Thomas, Brian C. AU - Barrangou, Rodolphe AU - Banfield, Jillian F. T2 - MBIO AB - Many bacteria rely on CRISPR-Cas systems to provide adaptive immunity against phages, predation by which can shape the ecology and functioning of microbial communities. To characterize the impact of CRISPR immunization on phage genome evolution, we performed long-term bacterium-phage (Streptococcus thermophilus-phage 2972) coevolution experiments. We found that in this species, CRISPR immunity drives fixation of single nucleotide polymorphisms that accumulate exclusively in phage genome regions targeted by CRISPR. Mutation rates in phage genomes highly exceed those of the host. The presence of multiple phages increased phage persistence by enabling recombination-based formation of chimeric phage genomes in which sequences heavily targeted by CRISPR were replaced. Collectively, our results establish CRISPR-Cas adaptive immunity as a key driver of phage genome evolution under the conditions studied and highlight the importance of multiple coexisting phages for persistence in natural systems.Phages remain an enigmatic part of the biosphere. As predators, they challenge the survival of host bacteria and archaea and set off an "arms race" involving host immunization countered by phage mutation. The CRISPR-Cas system is adaptive: by capturing fragments of a phage genome upon exposure, the host is positioned to counteract future infections. To investigate this process, we initiated massive deep-sequencing experiments with a host and infective phage and tracked the coevolution of both populations over hundreds of days. In the present study, we found that CRISPR immunity drives the accumulation of phage genome rearrangements (which enable longer phage survival) and escape mutations, establishing CRISPR as one of the fundamental drivers of phage evolution. DA - 2015/// PY - 2015/// DO - 10.1128/mbio.00262-15 VL - 6 IS - 2 SP - SN - 2150-7511 ER - TY - JOUR TI - Blood lipid distribution, aortic cholesterol concentrations, and selected inflammatory and bile metabolism markers in Syrian hamsters fed a standard breeding diet AU - Stephens, A. M. AU - Sanders, T. H. T2 - Journal of the American Association for Laboratory Animal Science DA - 2015/// PY - 2015/// VL - 54 IS - 4 SP - 353-358 ER - TY - JOUR TI - Black Currant Anthocyanins Attenuate Weight Gain and Improve Glucose Metabolism in Diet-Induced Obese Mice with Intact, but Not Disrupted, Gut Microbiome AU - Esposito, Debora AU - Damsud, Thanakorn AU - Wilson, Mickey AU - Grace, Mary H. AU - Strauch, Renee AU - Li, Xu AU - Lila, Mary Ann AU - Komarnytsky, Slavko T2 - JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY AB - Black currant (Ribes nigrum L.) is a rich source of anthocyanins; however, the relationship between their apparently limited bioavailability and significant protection against metabolic pathologies is poorly understood. This study examined the gastrointestinal distribution of black currant anthocyanins and their phenolic acid metabolites in lean and diet-induced obese mice with healthy and antibiotic-disrupted microbiomes. Daily consumption of low- or high-fat diet supplemented with 1% black currant powdered extract (32% anthocyanins) for 8 weeks reduced body weight gain and improved glucose metabolism only in mice with the intact gut microbiome. Administration of antibiotic cocktail resulted in a 16-25-fold increase (P < 0.001) in anthocyanin content of feces, and cyanidin-based anthocyanins showed the largest increase in fecal content upon disruption of gut microbiome (92.3 ± 16.3 vs 4719 ± 158 μg/g feces), indicating their high susceptibility to microbial degradation in the gut. A 3-fold enrichment (P < 0.05) in gallic over protocatechuic acid was observed in the jejunum of both intact and antibiotic-treated animals, suggesting that this effect was likely independent of their gut microbiome status. Taken together, the data clearly demonstrate that gut microbiome and the type of the anthocyanin aglycone moiety can alter the protective effect of anthocyanins against obesity and associated insulin resistance. DA - 2015/7/15/ PY - 2015/7/15/ DO - 10.1021/acs.jafc.5b00963 VL - 63 IS - 27 SP - 6172-6180 SN - 1520-5118 KW - gastrointestinal tract KW - gut microbiome KW - obesity KW - inflammation KW - functional food ER - TY - JOUR TI - An explanatory framework of teachers' perceptions of a positive mealtime environment in a preschool setting AU - Mita, Satoko C. AU - Gray, Samuel A. AU - Goodell, L. Suzanne T2 - APPETITE AB - Attending a preschool center may help preschoolers with growth and development that encourage a healthy lifestyle, including sound eating behaviors. Providing a positive mealtime environment (PME) may be one of the keys to fostering a child's healthy eating habits in the classroom. However, a specific definition of a PME, the components of a PME, or directions on how to create one have not been established. The purpose of this study, therefore, was to explore Head Start teachers' perceptions related to a PME and create a conceptual framework representing these perceptions. To achieve this purpose, researchers conducted 65 in-depth phone interviews with Head Start teachers around the US. Applying principles of grounded theory, researchers developed a conceptual framework depicting teachers' perceptions of PME, consisting of five key components: (1) the people (i.e., teachers, kitchen staff, parent volunteers, and children), (2) positive emotional tone (e.g., relaxed and happy), (3) rules, expectations, and routines (e.g., family-style mealtime), (4) operations of a PME (i.e., eating, socialization, and learning), and (5) both short- and long-term outcomes of a PME. With this PME framework, researchers may be able to enhance the effectiveness of nutrition interventions related to a PME, focusing on the factors in the conceptual framework as well as barriers associated with achieving these factors. DA - 2015/7/1/ PY - 2015/7/1/ DO - 10.1016/j.appet.2015.02.031 VL - 90 SP - 37-44 SN - 1095-8304 KW - Preschool teachers KW - Mealtime environment KW - Healthy eating KW - Grounded theory KW - Qualitative research ER - TY - JOUR TI - Alaskan seaweeds lower inflammation in RAW 264.7 macrophages and decrease lipid accumulation in 3T3-L1 adipocytes AU - Kellogg, Joshua AU - Esposito, Debora AU - Grace, Mary H. AU - Komarnytsky, Slavko AU - Lila, Mary Ann T2 - JOURNAL OF FUNCTIONAL FOODS AB - Chronic inflammation is characterized by macrophage accumulation in adipose tissue, which subsequently up-regulates pro-inflammatory cytokines and promotes the dysregulation of lipid metabolism, ultimately leading to insulin resistance. This study was designed to examine the effects of coastal Alaskan seaweeds on the macrophage inflammatory response and lipid metabolism of adipocytes. Two bioactive subfractions from the brown alga Fucus distichus, a monoglycosyldiacylglycerol subfraction and a phlorotannin subfraction, decreased mRNA expression of acute and chronic inflammatory biomarkers. Expression of Toll-like receptors TLR4 and TLR9 were also reduced, suggesting a potential mechanism of anti-inflammatory activity via TLR attenuation. F. distichus fractions decreased lipid accumulation up to 55% and increased free glycerol concentrations by 28–45%. This result was supported by increases in adiponectin and UCP-1 and decreases in leptin mRNA expression. Overall, the Alaskan seaweed F. distichus inhibited proinflammatory responses and improved lipid metabolism, suggesting the potential for seaweed phytochemicals to attenuate inflammatory diseases. DA - 2015/5// PY - 2015/5// DO - 10.1016/j.jff.2015.03.049 VL - 15 SP - 396-407 SN - 2214-9414 KW - Seaweed KW - Phlorotannin KW - Monoglycosyldiacylglycerol KW - Inflammation KW - Insulin resistance KW - Fucus distichus ER - TY - JOUR TI - Aerobic Hydrogen Production via Nitrogenase in Azotobacter vinelandii CA6 AU - Noar, Jesse AU - Loveless, Telisa AU - Luis Navarro-Herrero, Jose AU - Olson, Jonathan W. AU - Bruno-Barcena, Jose M. T2 - APPLIED AND ENVIRONMENTAL MICROBIOLOGY AB - The diazotroph Azotobacter vinelandii possesses three distinct nitrogenase isoenzymes, all of which produce molecular hydrogen as a by-product. In batch cultures, A. vinelandii strain CA6, a mutant of strain CA, displays multiple phenotypes distinct from its parent: tolerance to tungstate, impaired growth and molybdate transport, and increased hydrogen evolution. Determining and comparing the genomic sequences of strains CA and CA6 revealed a large deletion in CA6's genome, encompassing genes related to molybdate and iron transport and hydrogen reoxidation. A series of iron uptake analyses and chemostat culture experiments confirmed iron transport impairment and showed that the addition of fixed nitrogen (ammonia) resulted in cessation of hydrogen production. Additional chemostat experiments compared the hydrogen-producing parameters of different strains: in iron-sufficient, tungstate-free conditions, strain CA6's yields were identical to those of a strain lacking only a single hydrogenase gene. However, in the presence of tungstate, CA6 produced several times more hydrogen. A. vinelandii may hold promise for developing a novel strategy for production of hydrogen as an energy compound. DA - 2015/7// PY - 2015/7// DO - 10.1128/aem.00679-15 VL - 81 IS - 13 SP - 4507-4516 SN - 1098-5336 ER - TY - JOUR TI - Stability and immunogenicity of hypoallergenic peanut protein-polyphenol complexes during in vitro pepsin digestion AU - Plundrich, Nathalie J. AU - White, Brittany L. AU - Dean, Lisa L. AU - Davis, Jack P. AU - Foegeding, E. Allen AU - Lila, Mary Ann T2 - FOOD & FUNCTION AB - Allergenic peanut proteins are relatively resistant to digestion, and if digested, metabolized peptides tend to remain large and immunoreactive, triggering allergic reactions in sensitive individuals. In this study, the stability of hypoallergenic peanut protein-polyphenol complexes was evaluated during simulated in vitro gastric digestion. When digested with pepsin, the basic subunit of the peanut allergen Ara h 3 was more rapidly hydrolyzed in peanut protein-cranberry or green tea polyphenol complexes compared to uncomplexed peanut flour. Ara h 2 was also hydrolyzed more quickly in the peanut protein-cranberry polyphenol complex than in uncomplexed peanut flour. Peptides from peanut protein-cranberry polyphenol complexes and peanut protein-green tea polyphenol complexes were substantially less immunoreactive (based on their capacity to bind to peanut-specific IgE from patient plasma) compared to peptides from uncomplexed peanut flour. These results suggest that peanut protein-polyphenol complexes may be less immunoreactive passing through the digestive tract in vivo, contributing to their attenuated allergenicity. DA - 2015/// PY - 2015/// DO - 10.1039/c5fo00162e VL - 6 IS - 7 SP - 2145-2154 SN - 2042-650X ER - TY - JOUR TI - SIGNR3-dependent immune regulation by Lactobacillus acidophilus surface layer protein A in colitis AU - Lightfoot, Yaima L. AU - Selle, Kurt AU - Yang, Tao AU - Goh, Yong Jun AU - Sahay, Bikash AU - Zadeh, Mojgan AU - Owen, Jennifer L. AU - Colliou, Natacha AU - Li, Eric AU - Johannssen, Timo AU - Lepenies, Bernd AU - Klaenhammer, Todd R. AU - Mohamadzadeh, Mansour T2 - EMBO JOURNAL AB - Article9 February 2015free access SIGNR3-dependent immune regulation by Lactobacillus acidophilus surface layer protein A in colitis Yaíma L Lightfoot Yaíma L Lightfoot Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Kurt Selle Kurt Selle Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA Search for more papers by this author Tao Yang Tao Yang Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Yong Jun Goh Yong Jun Goh Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA Search for more papers by this author Bikash Sahay Bikash Sahay Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Mojgan Zadeh Mojgan Zadeh Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Jennifer L Owen Jennifer L Owen Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Natacha Colliou Natacha Colliou Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Eric Li Eric Li Division of Infectious Diseases and Global Medicine, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Timo Johannssen Timo Johannssen Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Search for more papers by this author Bernd Lepenies Bernd Lepenies Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Search for more papers by this author Todd R Klaenhammer Todd R Klaenhammer Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA Search for more papers by this author Mansour Mohamadzadeh Corresponding Author Mansour Mohamadzadeh Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Yaíma L Lightfoot Yaíma L Lightfoot Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Kurt Selle Kurt Selle Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA Search for more papers by this author Tao Yang Tao Yang Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Yong Jun Goh Yong Jun Goh Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA Search for more papers by this author Bikash Sahay Bikash Sahay Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Mojgan Zadeh Mojgan Zadeh Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Jennifer L Owen Jennifer L Owen Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Natacha Colliou Natacha Colliou Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Eric Li Eric Li Division of Infectious Diseases and Global Medicine, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Timo Johannssen Timo Johannssen Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Search for more papers by this author Bernd Lepenies Bernd Lepenies Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Search for more papers by this author Todd R Klaenhammer Todd R Klaenhammer Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA Search for more papers by this author Mansour Mohamadzadeh Corresponding Author Mansour Mohamadzadeh Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA Search for more papers by this author Author Information Yaíma L Lightfoot1,2, Kurt Selle3, Tao Yang1,2, Yong Jun Goh3, Bikash Sahay1,2, Mojgan Zadeh1,2, Jennifer L Owen4, Natacha Colliou1,2, Eric Li5, Timo Johannssen6,7, Bernd Lepenies6,7, Todd R Klaenhammer3 and Mansour Mohamadzadeh 1,2 1Department of Infectious Diseases and Pathology, University of Florida, Gainesville, FL, USA 2Division of Gastroenterology, Hepatology & Nutrition, Department of Medicine, University of Florida, Gainesville, FL, USA 3Department of Food, Bioprocessing and Nutrition Sciences, and Genomic Sciences Program, North Carolina State University, Raleigh, NC, USA 4Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL, USA 5Division of Infectious Diseases and Global Medicine, Department of Medicine, University of Florida, Gainesville, FL, USA 6Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany 7Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany *Corresponding author. Tel: +1 352 294 4117; E-mail: [email protected] The EMBO Journal (2015)34:881-895https://doi.org/10.15252/embj.201490296 See also: G Lugo-Villarino & O Neyrolles (April 2015) PDFDownload PDF of article text and main figures. Peer ReviewDownload a summary of the editorial decision process including editorial decision letters, reviewer comments and author responses to feedback. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Intestinal immune regulatory signals govern gut homeostasis. Breakdown of such regulatory mechanisms may result in inflammatory bowel disease (IBD). Lactobacillus acidophilus contains unique surface layer proteins (Slps), including SlpA, SlpB, SlpX, and lipoteichoic acid (LTA), which interact with pattern recognition receptors to mobilize immune responses. Here, to elucidate the role of SlpA in protective immune regulation, the NCK2187 strain, which solely expresses SlpA, was generated. NCK2187 and its purified SlpA bind to the C-type lectin SIGNR3 to exert regulatory signals that result in mitigation of colitis, maintenance of healthy gastrointestinal microbiota, and protected gut mucosal barrier function. However, such protection was not observed in Signr3−/− mice, suggesting that the SlpA/SIGNR3 interaction plays a key regulatory role in colitis. Our work presents critical insights into SlpA/SIGNR3-induced responses that are integral to the potential development of novel biological therapies for autoinflammatory diseases, including IBD. Synopsis Functional intestinal homeostasis is tightly controlled by the interaction of gut microbial gene products with pattern recognition receptors, including C-type lectins. Bacterial SlpA via interaction with SIGNR3, the human DC-SIGN ortholog, controls proinflammation by reducing critical proinflammatory signals, thereby allowing the differentiation of extrathymic regulatory T cells (Tregs) and the mitigation of colitis. Interruption of SlpA:SIGNR3 interaction leads to the dysfunction of intestinal homeostasis and the formation of proinflammatory innate cells and Tregs. The Lactobacillus acidophilus surface layer protein SlpA protects mice against T-cell-, DSS-, and infection-induced colitis. SlpA enhances the intestinal barrier and the gut microbiota, and regulates immune responses in steady state and in disease. SlpA binds to murine C-type lectins, SIGNR3, and its human ortholog DC-SIGN. Protection by SlpA is highly controlled by SIGNR3 signaling. Introduction The gastrointestinal (GI) microbiota plays a critical role in determining the immunologic outcome of various signaling events in host cells via their gene products, exceeding the human genome by a 100-fold (Ley et al, 2006; Qin et al, 2010). As such, the composition of the GI microbiota and host immunity are mutualistic and continuously influence each other (Maslowski & Mackay, 2011; McDermott & Huffnagle, 2014). Therefore, it is inevitable that intestinal homeostasis is tightly controlled by regulatory immune mechanisms, which are established by the interactions of the trillions of microbes and their gene products with numerous pattern recognition receptors (PRRs), including C-type lectin receptors (CLRs), such as the specific intracellular adhesion molecule-3 grabbing non-integrin homolog-related 3 (SIGNR3) (Konstantinov et al, 2008; Osorio & Reis e Sousa, 2011). Disruption of this delicate balance by inimical signals has devastating consequences that may result in intestinal disorders, including inflammatory bowel disease (IBD). When this occurs, highly activated innate cells trigger intestine-infiltrating pathogenic T-cell subsets (e.g. Th1, Th17), and even regulatory T cells (Tregs) with proinflammatory characteristics (Khazaie et al, 2012; Geremia et al, 2014; Neurath, 2014) that ultimately drive tissue destruction and intestinal disease progression. Innate cells (e.g. dendritic cells, macrophages) are the initial targets of the culpable microbes and their gene products, which subsequently affect the regulation/stimulation of intestinal immunity (Ivanov & Honda, 2012; Atarashi et al, 2013; Yang et al, 2014). Given these entwined relationships, it is not surprising that microbial products have been linked to the pathology of intestinal autoinflammation (Nicholson et al, 2012). The underlying associations between gut microbes and inflammatory diseases (e.g. IBD) have already been well documented; however, the cellular and molecular mechanisms by which intestinal commensal gene product(s) and their molecular receptor(s) impact immune responses remain unclear. Information regarding the immunobiologic functions of Lactobacillus acidophilus surface layer proteins (Slps) is relatively limited. Slps are paracrystalline (glyco) protein arrays that are abundant on the cell surfaces of few eubacteria and archaea, including L. acidophilus (Johnson et al, 2013). The S-layer of L. acidophilus NCFM is composed of three Slp-encoding genes: slpA (LBA0169), slpB (LBA0175), and slpX (LBA0512) (Goh et al, 2009). Diverse functional roles have been proposed for Slps, including cell shape determinants, molecular sieves, protective layers against viral infection, anchoring sites for surface-associated enzymes, and facilitators of cellular adhesion through PRRs, including C-type lectins (CLECs) (Konstantinov et al, 2008). CLECs recognize carbohydrate structures on self and nonself antigens (Engering et al, 2002; Osorio & Reis e Sousa, 2011). Twenty-nine CLECs, including DC-specific ICAM-3-grabbing non-integrin (DC-SIGN), have been identified on dendritic cells (DCs) and macrophages (MΦs) thus far (van Kooyk & Geijtenbeek, 2003; Ehlers, 2010; Sancho & Reis e Sousa, 2013). DC-SIGN, which was previously shown to bind L. acidophilus SlpA in vitro (Konstantinov et al, 2008), is a calcium-dependent carbohydrate-binding protein with specificity for the mannose-containing glycans of microbial surface components and fucose-containing Lewis antigens (Ehlers, 2010). Of the eight murine homologs of DC-SIGN, SIGNR3 (CD209d) exhibits the most biochemical similarity to human DC-SIGN (Powlesland et al, 2006). SIGNR3 contains a carbohydrate recognition domain (CRD) and signals through a hemi-immunoreceptor tyrosine-based activation motif (hemi-ITAM) (Tanne et al, 2009). Such signaling potentially downregulates the ubiquitously expressed leukotriene A4 hydrolase (LTA4H) (Tobin et al, 2010) that catalyzes proinflammatory leukotriene B4 (LTB4) synthesis from LTA4 (Snelgrove et al, 2010), which subsequently activates interleukin (IL)-1β production. Here, we identify SlpA as a key effector molecule expressed by L. acidophilus and demonstrate its in vivo protective role in murine colitis models. Moreover, we provide evidence that protection by L. acidophilus SlpA is conferred via signaling through a single CLR, namely SIGNR3. Results NCK2187 promotes intestinal immune regulation in steady state Recently, we showed that transient colonization of the colon with NCK2025 (LTA−) significantly mitigated chemical and T-cell-mediated colitis (Mohamadzadeh et al, 2011). Additionally, NCK2025 significantly abated inflammation-promoting polyposis in our novel Apclox468 × TS4-Cre mouse model, where protection correlated with the regulation of innate and T-cell-induced inflammation (Khazaie et al, 2012). We therefore hypothesized that the controlled inflammation resulted from cross talk between NCK2025 SlpA with intestinal cells. To test this hypothesis, the upp-counterselective gene replacement strategy (Goh et al, 2009) was used to generate in-frame deletions in the slpB and slpX genes of NCK2030. The LTA− derivative was created by a deletion of the phosphoglycerol transferase gene (Mohamadzadeh et al, 2011), resulting in NCK2187, which expresses only SlpA (Fig 1A and B). To demonstrate that the newly generated NCK2187 transiently colonizes the gut, the clearance kinetics of both the erythromycin-resistant NCK56 and NCK2187 strains were determined in C57BL/6 (B6) mice that were orally treated once with 109 CFU/mouse. Data show that mice cleared both NCK56 and NCK2187 after 3 days, indicating that the deletion of LTA, SlpB, and SlpX in NCK2187 did not alter its transient passage through the GI tract when compared to its WT parent (Fig 1C). Figure 1. Lactobacillus acidophilus NCK2187 strain development and characteristics Agarose gel image illustrating PCR amplicons of ltaS (LBA0447), slpB, and slpX deletions in NCK2187. SDS–PAGE gel of 5 M LiCl-purified S-layer fractions from the parental strains, NCK56 and NCK1909, NCK2030 (LTA+, SlpA+, SlpB−, SlpX−), and NCK2187 (LTA−, SlpA+, SlpB−, SlpX−). B6 mice were orally gavaged with 109 CFU erythromycin-resistant NCK56 or NCK2187. Fecal pellets were collected daily and tested for the presence of erythromycin-resistant strains. n = 3 mice/group. Data are representative of five independent experiments and are shown as mean ± SEM. Colonic LP cells were co-cultured with NCK56 or NCK2187 (1:1), and secreted cytokines were measured in the supernatants. Data are shown as mean ± SEM and are representative of two experiments performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001. Download figure Download PowerPoint To investigate the activation of colonic DCs when co-cultured with NCK56 or NCK2187, colonic cells were obtained from naïve B6 mice. While such intestinal cell–bacterial co-cultures did not significantly change the expression of DC co-stimulatory molecules (e.g. CD40) (not shown) or IL-10, only NCK56 elevated the levels of IL-1β, IL-6, IL-12, and TNF-α (Fig 1D). Next, naïve B6 mice were orally gavaged with NCK56 or NCK2187, and colonic immune responses were analyzed. Treatment with NCK2187 significantly increased the frequency of colonic FoxP3+ Tregs when compared to both untreated (PBS) and NCK56-treated mice (Fig 2A). Moreover, IL-17A+ and IFN-γ+ CD4+ T cells were significantly reduced by NCK2187 treatment (Fig 2A). NCK2187-treated B6 FoxP3-GFP mice also exhibited higher numbers of colonic IL-10+ TGF-β1+ Tregs than did NCK56-treated and untreated mice (Fig 2B and C). Collectively, oral treatment with this novel L. acidophilus strain induced colonic regulatory immune responses. Figure 2. Lactobacillus acidophilus NCK2187 promotes intestinal regulation in steady state A. B6 mice were orally gavaged with 109 CFU NCK56 (blue) or NCK2187 (green) on days 0, 3, 6, and 9, or left untreated, and immune responses in the colon analyzed at day 14 by flow cytometry. B, C. B6 FoxP3-GFP mice were treated and evaluated as in (A). Regulatory cytokine production in FoxP3-GFP+ (green-dotted bars) versus FoxP3-GFP− (white bars) cells was measured by intracellular staining and FACS analyses (C). Data information: For all data shown, n = 5 mice/group. Data represent four individual experiments and are shown as mean ± SEM. *P < 0.05, **P < 0.01, ****P < 0.0001. Black asterisks compare NCK2187 to untreated (PBS) mice, and red asterisks to NCK56-treated mice. Download figure Download PowerPoint Protective properties of NCK2187 and its SlpA against intestinal inflammation and dysbiosis To clarify the consequences of the immunoregulatory responses observed above during inflammation, B6 Rag1−/− mice adoptively transferred with CD45RBhi CD4+ T cells were orally treated with NCK56, NCK2187, its purified SlpA, or PBS (Fig 3). Untreated (PBS) and NCK56-treated mice with adoptively transferred T cells developed severe colitis as demonstrated by weight loss, bloody diarrhea, shortening of the colon, and increased damage of the colon (Fig 3A–C, Supplementary Fig S1A). Furthermore, the levels of systemically induced proinflammatory IL-1β, IL-6, TNF-α, IFN-γ, G-CSF, and macrophage inflammatory protein (MIP)-1α were significantly enhanced in the sera of these groups of mice (Fig 3D). In contrast, similar to the Treg co-transferred mice, NCK2187 and its purified SlpA significantly protected Rag1−/− mice from T-cell-induced colitis (Supplementary Tables S1, S2 and S3). NCK2187- and SlpA-treated mice gained weight throughout the course of the study and did not develop bloody diarrhea in the way that the PBS and NCK56 groups did (Fig 3A). Furthermore, cecal and colonic atrophy due to pathogenic inflammation was not observed in these mice, as the tissue destruction and immune cell infiltration associated with T-cell-induced colitis were significantly abrogated in NCK2187- and SlpA-treated groups (Fig 3B and C). Accordingly, systemic inflammation was significantly reduced in these groups of mice (Fig 3D). Interestingly, the genes encoding the receptors for LTB4, Ltb4r1 and Ltb4r2, were significantly downregulated in the colons of NCK2187- and SlpA-treated mice (Supplementary Fig S1B), which may have contributed to the reduced expression of colonic Il1b (Supplementary Fig S1B). Figure 3. Lactobacillus acidophilus NCK2187 and its SlpA protect against pathogenic T-cell-induced colitis A. B6 Rag1−/− mice were injected with 106 CD4+CD45RBhi T cells and then orally gavaged with NCK56 (red), NCK2187 (green), or SlpA (blue), 1 and 3 days after transfer, and subsequently once a week for four consecutive weeks, or left untreated (magenta). A group of mice was co-transferred with CD4+CD25+ T cells as a positive control for protection (Tregs; gray). Colitis severity was determined in part by weight loss, diarrhea scores, and FOB. See Supplementary Tables S1, S2 and S3 for statistical analyses results. B, C. Colitis scores based on histopathology and gross morphology of the colons of mice treated as in (A) were also used as measures of disease. Scale bar = 50 μm. D. Circulating levels of proinflammatory cytokines were measured in the sera of the mice transferred and treated as in (A), or sham adoptive transferred (white bars). Data information: Data represent three individual experiments and are shown as mean ± SEM. n = 5 mice/group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Black asterisks compare NCK2187 to PBS-treated adoptively transferred mice, and red asterisks to NCK56-treated mice. Download figure Download PowerPoint T-cell-induced colitis resulted in intestinal epithelial erosions and ulcerations in mice that did not receive NCK2187 or its purified SlpA (Fig 3C). Indeed, the colonic expression of tight junction-associated genes was significantly downregulated in PBS- and NCK56-treated Rag1−/− mice (Fig 4A). Furthermore, FITC-dextran permeability assays confirmed that these mice were suffering from a dysfunctional intestinal barrier (Fig 4B). Accordingly, NCK2187 and SlpA significantly protected barrier integrity and function (Fig 4A and B). An immunologically and anatomically weakened intestinal epithelial barrier during intestinal inflammation allows luminal bacteria to interact with the intestinal mucosae and the infiltrating immune cells, initiating inflammatory responses directed against the gut commensals and introducing dysbiosis. To determine the status and composition of the microbiota in the T-cell-induced colitis model (week 7), we analyzed the microbial communities in the colons of the different experimental groups and found that the severity of colitis was associated with significant changes in the microbiota (Fig 4C–E, Supplementary Table S4). UniFrac analyses revealed that fecal bacterial diversity in PBS- and NCK56-treated Rag1−/− mice was modified in such a way that these groups were found to cluster separately from each other and from the protected mice (Fig 4C). Conversely, SlpA-, NCK2187-, and Treg-treated groups clustered together and showed similar phyla distributions (Fig 4C and D). Induced colitis in PBS- and NCK56-treated groups resulted in a significant contraction of members of the Bacteroidetes phyla (Fig 4D). Additionally, the normally underrepresented Verrucomicrobia phyla were increased in these colitogenic mice (Fig 4D), suggesting a shift in the intestinal milieu and the substrates available in the inflamed colon, which may promote the growth of previously underrepresented microbial communities so that they dominate the population. Alterations in the microbial composition were also manifested at lower taxonomic levels: NCK2187-, SlpA-, and Treg-treated groups once again showed similar relative abundance and distribution of several unclassified genera (Fig 4E). Figure 4. Lactobacillus acidophilus NCK2187 and its SlpA protect intestinal barrier function and prevent dysbiosis in pathogenic T-cell-induced colitisColitis was induced in B6 Rag1−/− mice as described in Fig 3. A, B. Colonic expression of tight junction-associated genes Cldn3 and Ocln, determined by RT–PCR relative to 18S rRNA (A), as well as passive transepithelial absorption of FITC-dextran (B), were used as measures of epithelial barrier integrity. Sham adoptive transferred B6 Rag1−/− mice (white bars) were used as baseline controls in some cases. n = 5 mice/group. Data represent three individual experiments and are shown as mean ± SEM. *P < 0.05, **P < 0.01. Black asterisks compare NCK2187 to PBS-treated adoptively transferred mice, and red asterisks to NCK56-treated mice. C. UniFrac analyses were used to calculate distances between the microbial communities of the different samples (week 7), and three-dimensional scatter plots were generated by using principal coordinate analyses (PCoA). Gray dots = CD4+CD45RBhi T cells + PBS; yellow dots = CD4+CD45RBhi T cells + NCK56; green dots = CD4+CD45RBhi T cells + NCK2187; blue dots = CD4+CD45RBhi T cells + SlpA; red dots = CD4+CD45RBhi T cells + Tregs. Each dot represents the fecal microbiota data of an individual mouse. D. Changes in the relative abundance of different phyla were also analyzed. See Supplementary Table S4 for results of statistical analyses among the different groups. Dark blue = Actinobacteria; green = Bacteroidetes; red = Proteobacteria; yellow = Verrucomicrobia; aqua = Firmicutes; purple = Tenericutes; black = others. E. Comparison of microbial communities at family or order levels. The heat map depicts the relative value in individual mice. n = 5–6 mice/group. Download figure Download PowerPoint To further investigate the regulatory role of L. acidophilus SlpA, we then selected an experimentally infectious model using Citrobacter rodentium that results in a breach of the intestinal epithelial barrier, potentially orchestrated by unrestrained proinflammatory immune responses (Lebeis et al, 2009). Accordingly, data demonstrate that treatment with NCK2187 and its purified SlpA significantly accelerated pathogen clearance (Supplementary Fig S2A), resulting in reduced size of the draining lymph nodes (Supplementary Fig S2B), and decreased colonic IL-1β expression (Supplementary Fig S2C). Conversely, this trend was not observed in C. rodentium-infected mice that were treated with NCK56 or PBS, suggesting that L. acidophilus SlpA regulates induced proinflammation (e.g. IL-1β), which results in less colonic damage and bloody diarrhea (Supplementary Fig S2D and E). Histologic analyses of colonic mucosal damage (e.g. goblet cell loss, abnormal crypts) and inflammation with C. rodentium alone or C. rodentium plus NCK56 revealed increased lymphoplasmacytic with lesser but mildly increased neutrophilic infiltrates within the lamina propria and colonic submucosae, which were decreased in L. acidophilus SlpA-treated groups (Supplementary Fig S2D). Future studies are warranted to elucidate the cellular and molecular mechanisms involved in such regulatory protective intestinal responses upon GI pathogen challenge. Lactobacillus acidophilus SlpA binding to SIGNR3 promotes colonic regulatory immune responses Symbiotic bacteria and their gene products dictate the nature of innate responses via their sensing receptors (Ivanov & Honda, 2012; Yang et al, 2014); however, such stimulatory signals must be regulated by other receptors to avoid intestinal inflammation. As stated earlier, SIGNR3 exhibits the most biochemical similarity to human DC-SIGN. We screened all known murine SIGNR1–8 and found the Signr1 and Signr3 genes to be differentially activated in the colonic tissue of mice orally treated with NCK2187 (Fig 5A), prompting us to evaluate the binding of SlpA to SIGNR1 and SIGNR3. Subsequently, the corresponding extracellular domains of SIGNR1 and SIGNR3 were fused to the Fc portion of human IgG1 (SIGNR1-hFc, SIGNR3-hFc) and then transiently expressed in Chinese hamster ovary (CHO)-S cells (Eriksson et al, 2013) (Supplementary Fig S3). Data demonstrate that while expressed SIGNR3-hFc bound to purified SlpA coated onto charged ferrous beads, SIGNR1-hFc, Ctrl-hFc (control protein tagged with hFc), and the secondary rat anti-human Fc antibody alone did not, suggesting SlpA-binding specificity to SIGNR3 (Fig 5B). Additionally, we observed the binding of L. acidophilus SlpA to full-length SIGNR3 but not SIGNR1 expressed on CHO-S cells, indicating once again receptor-binding specificity to this bacterial protein (Fig 5C). Moreover, given that SIGNR3 was found to be the mouse ortholog of DC-SIGN, most closely resembling its human homolog in terms of ligand binding (Powlesland et al, 2006), in vitro binding assays confirmed that purified SlpA binds to DC-SIGN-Fc (Fig 5D), as well as to CHO-S cells expressing DC-SIGN (Fig 5E). Figure 5. Lactobacillus acidophilus NCK2187 and its SlpA bind to murine SIGNR3 to induce regulatory signals A. B6 mice were orally gavaged with 109 CFU NCK56 or NCK2187, and the colonic gene expression of C-type lectin receptors were measured by RT–PCR. Each box represents an individual mouse; n = 4. Data represent three individual experiments. B. Binding of SlpA to various hFc fusion proteins was analyzed by flow cytometry. Gray tinted line = SlpA-coated beads only; orange = SlpA-coated beads + secondary antibody; green = SlpA-coated beads + control fusion protein; blue = SlpA-coated beads + SIGNR1-hFc; red = SlpA-coated beads + SIGNR3-hFc. Binding assay results were confirmed five independent times. C. Binding of SlpA to SIGNR3, but not SIGNR1 expressed by CHO-S cells. Gray tinted line = untransfected CHO-S cells; blue = untr DA - 2015/4/1/ PY - 2015/4/1/ DO - 10.15252/embj.201490296 VL - 34 IS - 7 SP - 881-895 SN - 1460-2075 KW - colitis KW - immune regulation KW - Lactobacillus acidophilus KW - SIGNR3 KW - surface layer protein A ER - TY - JOUR TI - Getting my child to eat the right amount. Mothers' considerations when deciding how much food to offer their child at a meal AU - Johnson, Susan L. AU - Goodell, L. Suzanne AU - Williams, Kimberly AU - Power, Thomas G. AU - Hughes, Sheryl O. T2 - APPETITE AB - Feeding young children successfully requires parenting skills, trust that children will eat, and nutrition and child development knowledge to ensure that foods and the amounts offered are developmentally appropriate. Mothers are often responsible for determining how much food is offered to their children; however, the influences on mothers' decisions regarding how much to offer their children – their motivations, goals for feeding and child consumption – have not been investigated. Study aims included gathering qualitative data regarding mothers' decisional processes related to preparing a dinner meal plate for her preschooler. Low income mothers (n = 15 African American and n = 15 Latina mothers) were recruited from preschools in the Denver, CO metropolitan area to participate in semi-structured interviews. Mothers prepared a plate for their preschooler and were asked about influences on their decisions about portion sizes and their expectations for children's eating. Data were coded by 2 independent coders using a consensus coding process and analyzed by investigators. Three themes emerged: (1) portion sizes differ for children who are “good” eaters and “picky” eaters; (2) mothers know the “right amounts” to serve their child; and (3) mothers have emotional investments in their children's eating. Some influencing factors were child-centered (e.g. child's likes and dislikes, “picky” and “good” eaters, and foods previously eaten in the day) and some related to adult expectations and concerns (nutrient content and waste). Interventions focusing on portion size may be more effective if tailored to the mothers' perceptions regarding her child's eating characteristics. DA - 2015/5/1/ PY - 2015/5/1/ DO - 10.1016/j.appet.2014.12.004 VL - 88 SP - 24-32 SN - 1095-8304 KW - Portion KW - Child KW - Preschool KW - Obesity KW - Underweight KW - Mother ER - TY - JOUR TI - Effects of Acetic Acid and Arginine on pH Elevation and Growth of Bacillus licheniformis in an Acidified Cucumber Juice Medium AU - Yang, Zhenquan AU - Meng, Xia AU - Breidt, Frederick, Jr. AU - Dean, Lisa L. AU - Arritt, Fletcher M. T2 - JOURNAL OF FOOD PROTECTION AB - Bacillus licheniformis has been shown to cause pH elevation in tomato products having an initial pH below 4.6 and metabiotic effects that can lead to the growth of pathogenic bacteria. Because of this, the organism poses a potential risk to acidified vegetable products; however, little is known about the growth and metabolism of this organism in these products. To clarify the mechanisms of pH change and growth of B. licheniformis in vegetable broth under acidic conditions, a cucumber juice medium representative of a noninhibitory vegetable broth was used to monitor changes in pH, cell growth, and catabolism of sugars and amino acids. For initial pH values between pH 4.1 to 6.0, pH changes resulted from both fermentation of sugar (lowering pH) and ammonia production (raising pH). An initial pH elevation occurred, with starting pH values of pH 4.1 to 4.9 under both aerobic and anaerobic conditions, and was apparently mediated by the arginine deiminase reaction of B. licheniformis. This initial pH elevation was prevented if 5 mM or greater acetic acid was present in the brine at the same pH. In laboratory media, under favorable conditions for growth, data indicated that growth of the organism was inhibited at pH 4.6 with protonated acetic acid concentrations of 10 to 20 mM, corresponding to 25 to 50 mM total acetic acid; however, growth inhibition required greater than 300 mM citric acid (10-fold excess of the amount in processed tomato products) products under similar conditions. The data indicate that growth and pH increase by B. licheniformis may be inhibited by the acetic acid present in most commercial acidified vegetable products but not by the citric acid in many tomato products. DA - 2015/4// PY - 2015/4// DO - 10.4315/0362-028x.jfp-14-478 VL - 78 IS - 4 SP - 728-737 SN - 1944-9097 ER - TY - JOUR TI - Destruction of the Capsid and Genome of GII.4 Human Norovirus Occurs during Exposure to Metal Alloys Containing Copper AU - Manuel, C. S. AU - Moore, M. D. AU - Jaykus, L. A. T2 - APPLIED AND ENVIRONMENTAL MICROBIOLOGY AB - Human norovirus (HuNoV) represents a significant public health burden worldwide and can be environmentally transmitted. Copper surfaces have been shown to inactivate the cultivable surrogate murine norovirus, but no such data exist for HuNoV. The purpose of this study was to characterize the destruction of GII.4 HuNoV and virus-like particles (VLPs) during exposure to copper alloy surfaces. Fecal suspensions positive for a GII.4 HuNoV outbreak strain or GII.4 VLPs were exposed to copper alloys or stainless steel for 0 to 240 min and recovered by elution. HuNoV genome integrity was assessed by reverse transcription-quantitative PCR (RT-qPCR) (without RNase treatment), and capsid integrity was assessed by RT-qPCR (with RNase treatment), transmission electron microscopy (TEM), SDS-PAGE/Western blot analysis, and a histo-blood group antigen (HBGA) binding assay. Exposure of fecal suspensions to pure copper for 60 min reduced the GII.4 HuNoV RNA copy number by ∼3 log10 units when analyzed by RT-qPCR without RNase treatment and by 4 log10 units when a prior RNase treatment was used. The rate of reduction of the HuNoV RNA copy number was approximately proportional to the percentage of copper in each alloy. Exposure of GII.4 HuNoV VLPs to pure-copper surfaces resulted in noticeable aggregation and destruction within 240 min, an 80% reduction in the VP1 major capsid protein band intensity in 15 min, and a near-complete loss of HBGA receptor binding within 8 min. In all experiments, HuNoV remained stable on stainless steel. These results suggest that copper surfaces destroy HuNoV and may be useful in preventing environmental transmission of the virus in at-risk settings. DA - 2015/8// PY - 2015/8// DO - 10.1128/aem.00388-15 VL - 81 IS - 15 SP - 4940-4946 SN - 1098-5336 ER - TY - JOUR TI - Chemical Composition of the Essential Oils from Leaves of Edible (Arachis hypogaea L.) and Perennial (Arachis glabrata Benth.) Peanut Plants AU - Constanza, Karen AU - Tallury, Shyamalrau AU - Whaley, Jeffrey AU - Sanders, Timothy AU - Dean, Lisa T2 - JOURNAL OF ESSENTIAL OIL BEARING PLANTS AB - :Peanut or groundnut (Arachis hypogaea L.) is a valuable oilseed crop, but other than the seed, the rest of the plant is of minimal value. Plant material including the leaves is used as soil conditioner or as animal feed. Perennial peanut (Arachis glabrata Benth and Arachis pintoi Krapov & W.C. Greg) known as forage or rhizoma peanut produces few seeds, but is grown specifically as a forage, turf or ornamental plant. The leaves from the peanut plants of the cultivated variety, Bailey and the perennial varieties, Arblick, Arbrook, and Amarillo were freeze dried, essential oils were extracted by distillation and the chemical compositions were determined using gas chromatography and gas chromatography-mass spectrometry. Oil yield from the A. hypogaea leaves was 0.0063 % (w/w). The major components were 1-octen-3-ol (12.4 %), heneicosane (11.7 %), nonanal (10.9 %), 4-vinylguaiacol (6.4 %) and phytol (7.4 %). The yield from A. glabrata and A. pintoi leaves ranged from 0.0044 % to 0.0061 % (w/w) with the major components in common among the three varieties tested 1-octen-3-ol (40.5-44.9 %), β-linalool (5.0-8.9 %) and 4-vinylguaiacol (1.4-8.6 %). DA - 2015/5/4/ PY - 2015/5/4/ DO - 10.1080/0972060x.2014.961039 VL - 18 IS - 3 SP - 605-612 SN - 0976-5026 ER - TY - JOUR TI - Associations between Weather and Microbial Load on Fresh Produce Prior to Harvest AU - Ward, Michelle AU - Dhingra, Radhika AU - Remais, Justin V. AU - Chang, Howard H. AU - Johnston, Lynette M. AU - Jaykus, Lee-Ann AU - Leon, Juan T2 - JOURNAL OF FOOD PROTECTION AB - Contaminated produce causes approximately 1 million cases of foodborne illness and 1 billion dollars in damages to the U.S. economy annually. The environmental conditions, especially weather, that influence the inoculation, proliferation, and dispersal of microbial load on produce are not well understood. Using a mixed models approach, we examined the relationship of temperature and precipitation to microbial indicators of contamination on fresh produce on the farm over a week-long period prior to harvest. Between 2000 and 2002, we assayed for four microbial indicators of contamination (aerobic plate count, Enterococcus, total coliforms, and Escherichia coli) on 10 produce types in 15 fields in the southern United States. The sample collection times varied, with most occurring between January and May. We collected hourly weather data for the corresponding time period and location. Our results indicated that there was a significant association between the average daily temperature (20°C) and both log aerobic plate count (e.g., an increase of 0.074 log CFU/g [standard error {SE}, 0.023] per °C increase in weekly average temperature) and log Enterococcus (e.g., an increase of 0.15 log CFU/g [SE, 0.031] per °C increase in weekly average temperature) for approximately 5 days prior to sample collection. Daily total precipitation was significantly associated with log coliforms on 2 days (∼0.11 log CFU/g [SE, 0.06] per mm of precipitation) during the week-long lag period prior to harvest. Our results suggest that microbial indicator concentrations may increase as the temperature increases. Precipitation may have a positive but complex relationship with microbial indicators, as precipitation may create moist conditions conducive to bacterial growth, spread contamination onto the field, or wash contamination off of the plant. DA - 2015/4// PY - 2015/4// DO - 10.4315/0362-028x.jfp-14-381 VL - 78 IS - 4 SP - 849-854 SN - 1944-9097 ER - TY - JOUR TI - Using State Diagrams for Predicting Colloidal Stability of Whey Protein Beverages AU - Wagoner, Ty B. AU - Ward, Loren AU - Foegeding, E. Allen T2 - JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY AB - A method for evaluating aspects of colloidal stability of whey protein beverages after thermal treatment was established. Three state diagrams for beverages (pH 3-7) were developed representing protein solubility, turbidity, and macroscopic state after two ultrahigh-temperature (UHT) treatments. Key transitions of stability in the state diagrams were explored using electrophoresis and chromatography to determine aggregation propensities of β-lactoglobulin, α-lactalbumin, bovine serum albumin, and glycomacropeptide. The state diagrams present an overlapping view of high colloidal stability at pH 3 accompanied by high solubility of individual whey proteins. At pH 5, beverages were characterized by poor solubility, high turbidity, and aggregation/gelation of whey proteins with the exception of glycomacropeptide. Stability increased at pH 6, due to increased solubility of α-lactalbumin. The results indicate that combinations of state diagrams can be used to identify key regions of stability for whey protein containing beverages. DA - 2015/5/6/ PY - 2015/5/6/ DO - 10.1021/acs.jafc.5b00633 VL - 63 IS - 17 SP - 4335-4344 SN - 1520-5118 KW - whey protein isolate KW - stability KW - heat treatment KW - beverage KW - solubility KW - turbidity ER - TY - JOUR TI - Occurrence and Diversity of CRISPR-Cas Systems in the Genus Bifidobacterium AU - Briner, Alexandra E. AU - Lugli, Gabriele Andrea AU - Milani, Christian AU - Duranti, Sabrina AU - Turroni, Francesca AU - Gueimonde, Miguel AU - Margolles, Abelardo AU - Sinderen, Douwe AU - Ventura, Marco AU - Barrangou, Rodolphe T2 - PLOS ONE AB - CRISPR-Cas systems constitute adaptive immune systems for antiviral defense in bacteria. We investigated the occurrence and diversity of CRISPR-Cas systems in 48 Bifidobacterium genomes to gain insights into the diversity and co-evolution of CRISPR-Cas systems within the genus and investigate CRISPR spacer content. We identified the elements necessary for the successful targeting and inference of foreign DNA in select Type II CRISPR-Cas systems, including the tracrRNA and target PAM sequence. Bifidobacterium species have a very high frequency of CRISPR-Cas occurrence (77%, 37 of 48). We found that many Bifidobacterium species have unusually large and diverse CRISPR-Cas systems that contain spacer sequences showing homology to foreign genetic elements like prophages. A large number of CRISPR spacers in bifidobacteria show perfect homology to prophage sequences harbored in the chromosomes of other species of Bifidobacterium, including some spacers that self-target the chromosome. A correlation was observed between strains that lacked CRISPR-Cas systems and the number of times prophages in that chromosome were targeted by other CRISPR spacers. The presence of prophage-targeting CRISPR spacers and prophage content may shed light on evolutionary processes and strain divergence. Finally, elements of Type II CRISPR-Cas systems, including the tracrRNA and crRNAs, set the stage for the development of genome editing and genetic engineering tools. DA - 2015/7/31/ PY - 2015/7/31/ DO - 10.1371/journal.pone.0133661 VL - 10 IS - 7 SP - SN - 1932-6203 ER - TY - JOUR TI - Integration of Resonant Acoustic (R) mixing into thermal processing of foods: A comparison study against other in-container sterilization technologies AU - Batmaz, Ediz AU - Sandeep, K. P. T2 - JOURNAL OF FOOD ENGINEERING AB - A comparison study was carried out for thermal processing times and final product quality in three different modes of retort operation. Still retort simulation, processing with horizontal reciprocating agitation, and ResonantAcoustic® Mixing (RAM) integrated processing were used to process banana puree samples at different °Bx (degree Brix) values (ranging from 16.6 to 41.5). Thermal processing times ranged from 76.6 min to 90.7 min for still retort simulation, from 6.8 min to 76.5 min for processing with horizontal reciprocating agitation, and from 4.3 min to 15.4 min for RAM integrated processing. While a significant amount of reduction in thermal process times (up to 91%) was observed for processing with horizontal reciprocating agitation for banana puree samples with a °Bx value between 16.6 and 29.1, this effectiveness dropped sharply (down to 16%) for samples with higher °Bx values (33.2–41.5). For the RAM integrated processing, the applied mixing technology was found to be effective in reducing the thermal process times and provide high quality post-process products throughout the tested range of product viscosities. DA - 2015/11// PY - 2015/11// DO - 10.1016/j.jfoodeng.2015.06.013 VL - 165 SP - 124-132 SN - 1873-5770 KW - Thermal processing KW - ResonantAcoustic (R) mixing KW - Horizontal reciprocathig agitation KW - Minimal processing ER - TY - JOUR TI - Identifying Key Attributes for Protein Beverages AU - Oltman, A. E. AU - Lopetcharat, K. AU - Bastian, E. AU - Drake, M. A. T2 - JOURNAL OF FOOD SCIENCE AB - This study identified key attributes of protein beverages and evaluated effects of priming on liking of protein beverages. An adaptive choice-based conjoint study was conducted along with Kano analysis to gain insight on protein beverage consumers (n = 432). Attributes evaluated included label claim, protein type, amount of protein, carbohydrates, sweeteners, and metabolic benefits. Utility scores for levels and importance scores for attributes were determined. Subsequently, two pairs of clear acidic whey protein beverages were manufactured that differed by age of protein source or the amount of whey protein per serving. Beverages were evaluated by 151 consumers on two occasions with or without priming statements. One priming statement declared "great flavor," the other priming statement declared 20 g protein per serving. A two way analysis of variance was applied to discern the role of each priming statement. The most important attribute for protein beverages was sweetener type, followed by amount of protein, followed by type of protein followed by label claim. Beverages with whey protein, naturally sweetened, reduced sugar and ≥15 g protein per serving were most desired. Three consumer clusters were identified, differentiated by their preferences for protein type, sweetener and amount of protein. Priming statements positively impacted concept liking (P < 0.05) but had no effect on overall liking (P > 0.05). Consistent with trained panel profiles of increased cardboard flavor with higher protein content, consumers liked beverages with 10 g protein more than beverages with 20 g protein (6.8 compared with 5.7, P < 0.05). Protein beverages must have desirable flavor for wide consumer appeal. DA - 2015/6// PY - 2015/6// DO - 10.1111/1750-3841.12877 VL - 80 IS - 6 SP - S1383-S1390 SN - 1750-3841 KW - conjoint KW - consumer preference KW - Kano KW - priming KW - protein beverages ER - TY - JOUR TI - Generation and characterization of nucleic acid aptamers targeting the capsid P domain of a human norovirus GII.4 strain AU - Moore, Matthew D. AU - Escudero-Abarca, Blanca I. AU - Suh, Soo Hwan AU - Jaykus, Lee-Ann T2 - JOURNAL OF BIOTECHNOLOGY AB - Human noroviruses (NoV) are the leading cause of acute viral gastroenteritis worldwide. Significant antigenic diversity of NoV strains has limited the availability of broadly reactive ligands for design of detection assays. The purpose of this work was to produce and characterize single stranded (ss)DNA aptamers with binding specificity to human NoV using an easily produced NoV target—the P domain protein. Aptamer selection was done using SELEX (Systematic Evolution of Ligands by EXponential enrichment) directed against an Escherichia coli-expressed and purified epidemic NoV GII.4 strain P domain. Two of six unique aptamers (designated M1 and M6-2) were chosen for characterization. Inclusivity testing using an enzyme-linked aptamer sorbent assay (ELASA) against a panel of 14 virus-like particles (VLPs) showed these aptamers had broad reactivity and exhibited strong binding to GI.7, GII.2, two GII.4 strains, and GII.7 VLPs. Aptamer M6-2 exhibited at least low to moderate binding to all VLPs tested. Aptamers significantly (p < 0.05) bound virus in partially purified GII.4 New Orleans outbreak stool specimens as demonstrated by ELASA and aptamer magnetic capture (AMC) followed by RT-qPCR. This is the first demonstration of human NoV P domain protein as a functional target for the selection of nucleic acid aptamers that specifically bind and broadly recognize diverse human NoV strains. DA - 2015/9/10/ PY - 2015/9/10/ DO - 10.1016/j.jbiotec.2015.06.389 VL - 209 SP - 41-49 SN - 1873-4863 KW - SELEX KW - Aptamer KW - Norovirus KW - Molecular detection KW - Viral diagnostics ER - TY - JOUR TI - Controlling Listeria monocytogenes in Cold Smoked Salmon with the Antimicrobial Peptide Salmine AU - Cheng, Christopher AU - Arritt, Fletcher AU - Stevenson, Clinton T2 - JOURNAL OF FOOD SCIENCE AB - Listeria monocytogenes (LM) is a major safety concern for smoked salmon producers, as it can survive both the brining and smoking process in cold smoked salmon production. Salmine is a cationic antimicrobial peptide derived from the milt of salmon that has been shown to inhibit the growth of LM in vitro. Commercialization of this peptide would add value to a waste product produced when raising salmon. The purpose of this study was to determine the anti-listeria activity of salmine in smoked salmon by measuring the viable counts of LM over time. Cold smoked salmon was treated with a salmine solution or coated with agar or k-carrageenan films incorporating salmine to maintain a high surface concentration of the antimicrobial. Samples were then inoculated with approximately 1.0 × 10(3) cells of LM. The viable counts were then enumerated throughout 4 wk at 4 °C storage. It was found that 5 mg/g salmine delayed the growth of LM on smoked salmon. These samples had significantly (P < 0.05) lower LM counts than on the untreated samples on days 13 and 22. Edible films did not significantly (P > 0.05) improve the antimicrobial efficacy of salmine. The peptide combined with biopolymers also had lower antimicrobial activity in vitro when compared to salmine alone. These results suggest there is potential for salmine to be used as a natural hurdle to inhibit growth of LM due to post process contamination; however, future investigations for extending this effect throughout the shelf life of smoked salmon products are warranted. DA - 2015/6// PY - 2015/6// DO - 10.1111/1750-3841.12886 VL - 80 IS - 6 SP - M1314-M1318 SN - 1750-3841 KW - antimicrobial KW - Listeria monocytogenes KW - salmon KW - smoked ER - TY - JOUR TI - Control of Histamine-Producing Bacteria and Histamine Formation in Fish Muscle by Trisodium Phosphate AU - Bjornsdottir-Butler, Kristin AU - Green, David P. AU - Bolton, Greg E. AU - McClellan-Green, Patricia D. T2 - JOURNAL OF FOOD SCIENCE AB - Scombrotoxin fish poisoning remains the primary cause of seafood poisoning outbreaks despite preventive guidelines. The purpose of this study was to investigate the use of pH for the control of growth and histamine formation by histamine-producing bacteria in fish muscle. We examined pH effects on growth and histamine formation in tuna fish infusion broth and in inoculated tuna and mahi-mahi fish muscle. Histamine production was significantly less for all bacterial strains at pH 8.5 compared to pH 5.5 in tuna fish infusion broth with no significant difference in growth. Elevated pH due to phosphate treatment of fish muscle tissues significantly reduced histamine formation with no effect on the growth of histamine-producing bacteria. This study revealed that phosphate treatment of mahi-mahi and tuna fish muscle resulted in significantly lower histamine production over 4 d of storage at 10 °C. Phosphate treatment of fish muscle may serve as a secondary barrier in addition to FDA recommended time and temperature controls for reducing public health concerns of scombrotoxin fish poisoning. DA - 2015/6// PY - 2015/6// DO - 10.1111/1750-3841.12875 VL - 80 IS - 6 SP - M1253-M1258 SN - 1750-3841 KW - bacteria KW - barrier KW - fish KW - histamine KW - histidine decarboxylase KW - pH ER - TY - JOUR TI - Consumer Acceptability of Cucumber Pickles Produced by Fermentation in Calcium Chloride Brine for Reduced Environmental Impact AU - Wilson, Emily M. AU - Johanningsmeier, Suzanne D. AU - Osborne, Jason A. T2 - JOURNAL OF FOOD SCIENCE AB - Fermentation of cucumbers in calcium chloride (CaCl2 ) brine has been proposed as an alternative process to reduce the environmental impact of traditional, high salt fermentations. The objective of this research was to determine whether consumer acceptability of pickle products would be impacted by fermentation and storage of cucumbers in CaCl2 brine. Cucumbers were fermented and stored with 0.1M CaCl2 or 1M sodium chloride (NaCl) in open-air, 3000 gal tanks at a commercial facility and processed into hamburger dill chips containing 0.38M NaCl. Cucumbers fermented in CaCl2 required additional desalting to reduce CaCl2 concentrations to that of current products. Consumers (n = 101) showed no significant preference for pickles from different fermentation treatments, whether stored for 2 mo (P = 0.75) or 8 mo (P = 0.68) prior to processing. In contrast, NaCl fermented pickles were preferred over CaCl2 fermented pickles stored for 10 mo and desalted only once (P < 0.01). A series of preference tests indicated that the taste of CaCl2 was not the factor affecting consumer preference, and the 50% detection threshold of CaCl2 in dill pickle chips was found to be 61.8 ± 7.6 mM, indicating that processors could potentially use CaCl2 fermentations with a single desalting step. Consumer liking of flavor (n = 73) was not influenced by fermentation in CaCl2 or by 23 or 35 mM CaCl2 in finished products (P > 0.05), but variability in texture decreased consumer liking (P < 0.05). Although promising, individual fermentation variability and texture quality of CaCl2 fermented products should be further evaluated prior to broad implementation of this process. DA - 2015/6// PY - 2015/6// DO - 10.1111/1750-3841.12882 VL - 80 IS - 6 SP - S1360-S1367 SN - 1750-3841 KW - calcium chloride taste KW - cucumber pickles KW - detection threshold KW - sodium-free fermentation KW - sustainability KW - vegetable preservation ER - TY - JOUR TI - Community-based research as a mechanism to reduce environmental health disparities in American Indian and Alaska native communities AU - McOliver, C. A. AU - Camper, A. K. AU - Doyle, J. T. AU - Eggers, M. J. AU - Ford, T. E. AU - Lila, M. A. AU - Berner, J. AU - Campbell, L. AU - Donatuto, J. T2 - International Journal of Environmental Research and Public Health DA - 2015/// PY - 2015/// VL - 12 IS - 4 SP - 4076-4100 ER - TY - JOUR TI - Combined use of variable pressure scanning electron microscopy and confocal laser scanning microscopy best reveal microstructure of comminuted meat gels AU - Liu, Wenjie AU - Lanier, Tyre C. T2 - LWT-FOOD SCIENCE AND TECHNOLOGY AB - Observing the distribution of protein and fat phases in comminuted meat products can be helpful to understanding the mechanisms of texture development and fat/water binding. In this study variable pressure scanning electron microscopy (VP-SEM) was compared to conventional scanning electron microscopy (SEM), and contrasted with confocal laser scanning microcopy (CLSM), as tools to characterize gel morphology of cooked meat batters or non-fat pastes. Gel morphology was varied by inclusion of whey protein isolate (WPI) that gels only at high temperature, in partial substitution of myofibrillar protein (MFP). CLSM (magnification:10–1000×) revealed that, when no WPI was added, a homogeneous gel structure was produced enmeshing small, well-distributed fat particles. Substituting 30 g/100 g MFP by WPI produced a coarse gel structure with clear microphase separation of fat. VP-SEM (magnification:1000–2000×) enabled visualization of small pore structure of gel matrix whereas SEM obscured details of this, as well as of the relationship between fat globules and gel matrix that were visible by VP-SEM. Since meat gels properties can be affected by multiple morphological features, visible only at different levels of magnification, the relationships between microstructure and important properties of meat gels can be most advantageously observed when both VP-SEM and CLSM are used in tandem. DA - 2015/7// PY - 2015/7// DO - 10.1016/j.lwt.2015.02.001 VL - 62 IS - 2 SP - 1027-1033 SN - 1096-1127 KW - Variable pressure scanning electron microscopy KW - Confocal laser scanning microscopy KW - Conventional scanning electron microscopy KW - Comminuted meat gels KW - Microstructure ER - TY - JOUR TI - Advances in CRISPR-Cas9 genome engineering: lessons learned from RNA interference AU - Barrangou, Rodolphe AU - Birmingham, Amanda AU - Wiemann, Stefan AU - Beijersbergen, Roderick L. AU - Hornung, Veit AU - Smith, Anja van Brabant T2 - NUCLEIC ACIDS RESEARCH AB - The discovery that the machinery of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 bacterial immune system can be re-purposed to easily create deletions, insertions and replacements in the mammalian genome has revolutionized the field of genome engineering and re-invigorated the field of gene therapy. Many parallels have been drawn between the newly discovered CRISPR-Cas9 system and the RNA interference (RNAi) pathway in terms of their utility for understanding and interrogating gene function in mammalian cells. Given this similarity, the CRISPR-Cas9 field stands to benefit immensely from lessons learned during the development of RNAi technology. We examine how the history of RNAi can inform today's challenges in CRISPR-Cas9 genome engineering such as efficiency, specificity, high-throughput screening and delivery for in vivo and therapeutic applications. DA - 2015/4/20/ PY - 2015/4/20/ DO - 10.1093/nar/gkv226 VL - 43 IS - 7 SP - 3407-3419 SN - 1362-4962 ER - TY - JOUR TI - Zero Risk Does Not Exist: Lessons Learned from Microbial Risk Assessment Related to Use of Water and Safety of Fresh Produce AU - De Keuckelaere, Ann AU - Jacxsens, Liesbeth AU - Amoah, Philip AU - Medema, Gertjan AU - McClure, Peter AU - Jaykus, Lee-Ann AU - Uyttendaele, Mieke T2 - COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY AB - Abstract Risk assessments related to use of water and safety of fresh produce originate from both water and food microbiology studies. Although the set‐up and methodology of risk assessment in these 2 disciplines may differ, analysis of the current literature reveals some common outcomes. Most of these studies from the water perspective focus on enteric virus risks, largely because of their anticipated high concentrations in untreated wastewater and their resistance to common wastewater treatments. Risk assessment studies from the food perspective, instead, focus mainly on bacterial pathogens such as Salmonella and pathogenic Escherichia coli . Few site‐specific data points were available for most of these microbial risk assessments, meaning that many assumptions were necessary, which are repeated in many studies. Specific parameters lacking hard data included rates of pathogen transfer from irrigation water to crops, pathogen penetration, and survival in or on food crops. Data on these factors have been investigated over the last decade and this should improve the reliability of future microbial risk estimates. However, the sheer number of different foodstuffs and pathogens, combined with water sources and irrigation practices, means that developing risk models that can span the breadth of fresh produce safety will be a considerable challenge. The new approach using microbial risk assessment is objective and evidence‐based and leads to more flexibility and enables more tailored risk management practices and guidelines. Drawbacks are, however, capacity and knowledge to perform the microbial risk assessment and the need for data and preferably data of the specific region. DA - 2015/7// PY - 2015/7// DO - 10.1111/1541-4337.12140 VL - 14 IS - 4 SP - 387-410 SN - 1541-4337 KW - fruits and vegetables KW - health risk KW - mitigation strategies KW - quantitative microbial risk assessment KW - water ER - TY - JOUR TI - Novel strategies for capturing health-protective mango phytochemicals in shelf stable food matrices AU - Guzman, Ivette AU - Grace, Mary H. AU - Yousef, Gad G. AU - Raskin, Ilya AU - Lila, Mary Ann T2 - INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION AB - Cost-effective methods for concentration and stabilization of otherwise perishable mango fruit phytoactives into shelf stable high protein ingredients were developed to combat stunting (malnutrition) in rural Africa. Mango juices complexed with sunflower oil and protein-rich legume flours yielded carotenoid-enriched oils and pelleted polyphenol-enriched flour matrices. Carotenoids from juices were concentrated 9-10 times in the fortified sunflower oil. Protein-rich soy and peanut flours captured 2.2-3.2 mg/g polyphenols from the juices. Alternatively, mango juice was sorbed and co-dried with flours, which stably bound the polyphenols, carotenoids, and natural sugars in soy or peanut protein-rich matrices. The concentration of provitamin A carotenoids was almost doubled and total polyphenols were enriched 4-5 times higher in the matrices compared to fresh pureed juice. Both strategies require minimal instrumentation, are compatible with rural village dietary practices; and capture the benefits of otherwise perishable seasonal resources by complexing healthful proteins together with phytoactive compounds. DA - 2015/3// PY - 2015/3// DO - 10.3109/09637486.2014.979315 VL - 66 IS - 2 SP - 175-185 SN - 1465-3478 KW - Carotenoids KW - fortified foods KW - polyphenols KW - protein-rich flour KW - vitamin A ER - TY - JOUR TI - Microbial Hazards in Irrigation Water: Standards, Norms, and Testing to Manage Use of Water in Fresh Produce Primary Production AU - Uyttendaele, Mieke AU - Jaykus, Lee-Ann AU - Amoah, Philip AU - Chiodini, Alessandro AU - Cunliffe, David AU - Jacxsens, Liesbeth AU - Holvoet, Kevin AU - Korsten, Lise AU - Lau, Mathew AU - McClure, Peter AU - Medema, Gertjan AU - Sampers, Imca AU - Jasti, Pratima Rao T2 - COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY AB - Abstract Accessibility to abundant sources of high‐quality water is integral to the production of safe and wholesome fresh produce. However, access to safe water is becoming increasingly difficult in many parts of the world, and this can lead to the production of fresh produce contaminated with pathogenic microorganisms, resulting in increased risk of human disease. Water, an important raw material in the fresh produce chain, is used in considerable amounts in many operations, including irrigation and application of pesticides and fertilizers, but also as a transport medium and for cooling and washing in postharvest practices. In several reported outbreaks related to uncooked fruit and vegetable products, water has been identified as a likely source of the outbreak. The present study, initiated by the ILSI Europe Emerging Microbiological Issues Task Force in collaboration with 8 other ILSI branches and support of WHO/FAO, was undertaken to review the status of, and provide suggestions for, consideration by different stakeholders on water and sanitation and its impact on food safety and public health. A limited number of guidelines and regulations on water quality for agricultural production are available, and many of them are still heavily based on microbial standards and (debated) parameters such as fecal coliforms. Data gaps have been identified with regard to baseline studies of microbial pathogens in water sources in many regions, the need for agreement on methods and microbial parameters to be used in assessing water quality, the fate of pathogens in water, and their transfer and persistence on irrigated/processed produce. DA - 2015/7// PY - 2015/7// DO - 10.1111/1541-4337.12133 VL - 14 IS - 4 SP - 336-356 SN - 1541-4337 KW - water KW - irrigation KW - outbreaks KW - fresh produce KW - good practices KW - testing ER - TY - JOUR TI - Unraveling the potential of CRISPR-Cas9 for gene therapy AU - Barrangou, Rodolphe AU - May, Andrew P. T2 - EXPERT OPINION ON BIOLOGICAL THERAPY AB - The molecular machinery from the prokaryotic clustered regularly interspaced short palindromic repeats (CRISPR)-Cas immune system has broadly been repurposed for genome editing in eukaryotes. In particular, the sequence-specific Cas9 endonuclease can be flexibly harnessed for the genesis of precise double-stranded DNA breaks, using single guide RNAs that are readily programmable. The endogenous DNA repair machinery subsequently generates genome modifications, either by random insertion or deletions using non-homologous end joining (NHEJ), or designed integration of mutations or genetic material using homology-directed repair (HDR) templates. This technology has opened new avenues for the investigation of genetic diseases in general, and for gene therapy applications in particular. DA - 2015/3// PY - 2015/3// DO - 10.1517/14712598.2015.994501 VL - 15 IS - 3 SP - 311-314 SN - 1744-7682 KW - Cas9 KW - CRISPR KW - gene therapy KW - genome editing ER - TY - JOUR TI - The roles of CRISPR-Cas systems in adaptive immunity and beyond AU - Barrangou, Rodolphe T2 - CURRENT OPINION IN IMMUNOLOGY AB - Clustered regularly interspaced short palindromic repeats (CRISPR) and accompanying Cas proteins constitute the adaptive CRISPR-Cas immune system in bacteria and archaea. This DNA-encoded, RNA-mediated defense system provides sequence-specific recognition, targeting and degradation of exogenous nucleic acid. Though the primary established role of CRISPR-Cas systems is in bona fide adaptive antiviral defense in bacteria, a growing body of evidence indicates that it also plays critical functional roles beyond immunity, such as endogenous transcriptional control. Furthermore, benefits inherent to maintaining genome homeostasis also come at the cost of reduced uptake of beneficial DNA, and preventing strategic adaptation to the environment. This opens new avenues for the investigation of CRISPR-Cas systems and their functional characterization beyond adaptive immunity. DA - 2015/2// PY - 2015/2// DO - 10.1016/j.coi.2014.12.008 VL - 32 SP - 36-41 SN - 1879-0372 ER - TY - JOUR TI - The Intricacies of Induced Lactation for Same-Sex Mothers of an Adopted Child AU - Wilson, Erica AU - Perrin, Maryanne Tigchelaar AU - Fogleman, April AU - Chetwynd, Ellen T2 - JOURNAL OF HUMAN LACTATION AB - The definition of a modern family is changing. In this case study, we describe the breastfeeding experience of a child receiving human milk from all 3 of his mothers: his 2 adoptive mothers, who induced lactation to nurse him, and his birth mother, who shared in his early feeding during the open adoption process and continued to pump and send milk to him for several months. We review the lactation protocol used by his adoptive mothers and the unique difficulties inherent in this multi-mother family dynamic. Both adoptive mothers successfully induced moderate milk production using a combination of hormonal birth control, domperidone, herbal supplements, and a schedule of breast pumping. However, because of the increased complexity of the immediate postpartum period and concerns with defining parental roles in a same-sex marriage, maintenance of milk production was difficult. DA - 2015/2// PY - 2015/2// DO - 10.1177/0890334414553934 VL - 31 IS - 1 SP - 64-67 SN - 1552-5732 KW - adoption KW - breastfeeding KW - induced lactation KW - lesbian ER - TY - JOUR TI - Escherichia coli O157:H7 bacteriophage Phi 241 isolated from an industrial cucumber fermentation at high acidity and salinity AU - Lu, Zhongjing AU - Breidt, Fred T2 - FRONTIERS IN MICROBIOLOGY AB - A novel phage, Φ241, specific for Escherichia coli O157:H7 was isolated from an industrial cucumber fermentation where both acidity (pH ≤ 3.7) and salinity (≥5% NaCl) were high. The phage belongs to the Myoviridae family. Its latent period was 15 min and average burst size was 53 phage particles per infected cell. The phage was able to lyse 48 E. coli O157:H7 strains, but none of the 18 non-O157 strains (including E. coli O104:H7) or the 2 O antigen-negative mutants of O157:H7 strain, 43895Δper (also lacking H7 antigen) and F12 (still expressing H7 antigen). However, the phage was able to lyse a per-complemented strain (43895ΔperComp) which expresses O157 antigen. These results indicated that phage Φ241 is specific for O157 antigen, and E. coli strains lacking O157 antigen were resistant to the phage infection, regardless of the presence or absence of H7 antigen. SDS-PAGE profile revealed at least 13 structural proteins of the phage. The phage DNA was resistant to many commonly used restriction endonucleases, suggesting the presence of modified nucleotides in the phage genome. At the multiplicity of infection of 10, 3, or 0.3, the phage caused a rapid cell lysis within 1 or 2 h, resulting in 3.5- or 4.5-log-unit reduction in cell concentration. The high lytic activity, specificity and tolerance to low pH and high salinity make phage Φ241 a potentially ideal biocontrol agent of E. coli O157:H7 in various foods. To our knowledge, this is the first report on E. coli O157:H7 phage isolated from high acidity and salinity environment. DA - 2015/2/17/ PY - 2015/2/17/ DO - 10.3389/fmicb.2015.00067 VL - 6 SP - SN - 1664-302X KW - bacteriophage KW - phage Phi 241 KW - Escherichia coli O157:H7 KW - biocontrol agent KW - cucumber fermentation KW - high acidity and salinity KW - food safety ER - TY - JOUR TI - Electromyographic Evaluation of Masticatory Muscles in Dentate Patients Versus Conventional and Implant-Supported Fixed and Removable Denture Wearers- A Preliminary Report Comparing Model Foods AU - Uram-Tuculescu, Sorin AU - Cooper, Lyndon F. AU - Foegeding, E. Allen AU - Vinyard, Christopher J. AU - De Kok, Ingeborg J. AU - Essick, Gregory T2 - INTERNATIONAL JOURNAL OF PROSTHODONTICS AB - To evaluate differences in masticatory muscle function during chewing of model foods designed to differ in fracture strength between dentate subjects (n = 5, ages 59 to 68 years) versus patients treated with a maxillary conventional complete denture opposing natural dentition or one of the following types of mandibular complete dentures: conventional, implant-supported overdenture, implant-supported fixed denture (n = 20, ages 45 to 83 years). The authors hypothesized that denture wearers would differ in duration of chewing, frequency of chewing, and masticatory muscle activity while preparing a bolus for swallowing.Surface electromyography was recorded bilaterally from the masseter, anterior temporalis, and anterior digastric. Masticatory muscle activity was evaluated using scaled values of the area under the electromyographic curve, while subjects chewed agar-based model foods with different fracture strengths. Chewing duration and frequency also were calculated from electromyographic recordings. Mixed model analysis of variance with "subject" as a random factor was used during statistical analysis. Logarithmic transformation was required to achieve normalization of residuals for the duration of chewing and the relative masticatory muscles activity, but not for the chewing frequency.Relative masticatory muscle activity was 2.57 times higher for the denture wearers than for the dentate subjects during chewing of model foods (P < .0001). The reduction in masticatory muscle activity from the 1st to the 10th chewing cycle was proportionally less in magnitude and occurred more gradually for denture wearers compared to dentate subjects. While chewing sequence duration increased with food fracture strength, it did not differ significantly in treatment versus dentate groups. Chewing cycle frequency did not differ between groups or with food fracture strength.The observed increases in relative masticatory muscle activity for denture wearers compared to the dentate subjects during oral food processing likely reflect supplemental mechanical efforts to accommodate the use of dentures for preparing a bolus for swallowing. DA - 2015/// PY - 2015/// DO - 10.11607/ijp.3931 VL - 28 IS - 1 SP - 79-92 SN - 1942-4426 ER - TY - JOUR TI - Bacteriophage exclusion, a new defense system AU - Barrangou, Rodolphe AU - Oost, John T2 - EMBO JOURNAL AB - Have you seen?12 December 2014free access Bacteriophage exclusion, a new defense system Rodolphe Barrangou Rodolphe Barrangou Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA Search for more papers by this author John van der Oost John van der Oost [email protected] Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands Search for more papers by this author Rodolphe Barrangou Rodolphe Barrangou Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA Search for more papers by this author John van der Oost John van der Oost [email protected] Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands Search for more papers by this author Author Information Rodolphe Barrangou1 and John Oost2 1Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC, USA 2Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands The EMBO Journal (2015)34:134-135https://doi.org/10.15252/embj.201490620 See also: T Goldfarb et al (January 2015) PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info The ability to withstand viral predation is critical for survival of most microbes. Accordingly, a plethora of phage resistance systems has been identified in bacterial genomes (Labrie et al, 2010), including restriction-modification systems (R-M) (Tock & Dryden, 2005), abortive infection (Abi) (Chopin et al, 2005), Argonaute-based interference (Swarts et al, 2014), as well as clustered regularly interspaced short palindromic repeats (CRISPR) and associated protein (Cas) adaptive immune system (CRISPR-Cas) (Barrangou & Marraffini, 2014; Van der Oost et al, 2014). Predictably, the dark matter of bacterial genomes contains a wealth of genetic gold. A study published in this issue of The EMBO Journal by Goldfarb et al (2015) unveils bacteriophage exclusion (BREX) as a novel, widespread bacteriophage resistance system that provides innate immunity against virulent and temperate phage in bacteria. It is well documented that viruses engage in a continuous co-evolutionary arms race with their microbial hosts, which has generated a broad arsenal of phage resistance mechanisms that are, in different combinations, widespread in bacteria and archaea. Actually, this evolutionary dialogue has shaped the genomic trajectory of microbial chromosomes, and previous studies have shown that phage resistance mechanisms often cluster within genomic defense islands (Makarova et al, 2011). One such defense island includes pglZ, which has been implicated in phage resistance as defined by phage growth limitation (Pgl) in Streptomyces coelicolor (Chinenova et al, 1982). The Pgl genotype consists of pglWXYZ, an operon encoding a serine/threonine kinase (PglW), an adenine-specific methyltransferase (PglX), an ATP-binding P-loop protein (PglY) and an alkaline phosphatase (PglZ). Investigating the occurrence of PglZ-containing cassettes in the genomes of ~1,500 bacteria and archaea, Goldfarb and collaborators establish that this phage resistance system is phylogenetically widespread, subject to horizontal gene transfer, and occurs in approximately 10% of microbial genomes. In the majority of cases, pglZ appears in a six-gene cassette (which they call BREX), which includes pglZ itself (phosphatase), pglX (methyltransferase) as well as brxABCL, that encodes an RNA-binding anti-termination protein (BrxA), an unknown protein (BrxB), an ATP-binding protein (BrxC) and a protease (BrxL). Further comparative analyses established six major BREX types based on gene sequence, content and number, all containing pglZ. In order to establish a direct correlation between the BREX genotype and a phage resistance phenotype, the authors insert the Bacillus cereus BREX system (Fig 1) into the Bacillus subtilis chromosome and confirm functional transcription in two operons (brxABC-pglX and pglZ-brxL). Next, they use a range of lytic and lysogenic phages for viral infection. Results show up to 10−5 reduction in efficiency of plaquing, illustrating a relatively high level of BREX-dependent phage resistance against both virulent and temperate phages. Figure 1. Bacteriophage exclusion(A) Invasion of a bacterial host by a temperate phage (left, red) and a lytic phage (purple, right). In the absence of an effective defense response, invasion will result in integration of the viral DNA in the host chromosome, whereas the DNA of the lytic viruses will be transcribed, translated and replicated, eventually leading to a new generation of viruses and lysis of the host cell. (B) Bacterial hosts that possess the bacteriophage exclusion (BREX) system are resistant to many (but not all) temperate and lytic phages. Different types of BREX systems are encoded by conserved gene clusters (in case of type-1 BREX from Bacillus cereus, two operons: brxABC-pglX and then pglZ-brxL). The well-conserved core of the BREX system includes three enzymes with (predicted) functionality: an ATP-binding P-loop protein (PglY/BrxC, C), an alkaline phosphatase (PglZ, Z) and a methyl-transferase (PglX, X) that specifically methylates TAGGAG. The B. cereus BREX system has been demonstrated to inhibit the integration of temperate phages, as well as the replication (and proliferation) of lytic phages (Goldfarb et al, 2015). Download figure Download PowerPoint Mechanistically, the authors reveal that BREX is distinct from the canonical Pgl system, as it provides phage resistance prior to the first round of infection. Because neither the release of newly generated phages, nor the integration of phage into the host chromosome was detected, they conclude that the BREX system provides resistance against both lytic and lysogenic phages. The authors rule out abortive infection and also show that BREX does not prevent phage adsorption, but rather blocks phage DNA replication. Remarkably, using Pac-Bio sequencing, the authors demonstrate that the host DNA is widely m6A methylated at the fifth position of a non-palindromic 5′-TAGGAG-3′ hexamer motif, whereas the phage DNA is not, perhaps allowing the host to distinguish viral from chromosomal DNA and presumably prevent self targeting. Indeed, deletion of the pglX gene that encodes a DNA methylase results in abrogated resistance, strongly suggesting that methylation of the host chromosome is required for BREX-mediated viral defense. Southern blot analyses of total cellular DNA reveal replication of phage DNA in BREX-lacking cells, but not in BREX-encoding cells. The authors exclude a mechanism of action akin to R-M, as there was no sign of phage DNA cleavage or degradation, suggesting that BREX is involved in inhibiting phage replication and propagation rather than degradation of viral DNA. The authors also test whether BREX can confer resistance against plasmid DNA uptake; results indicate that there is a mild effect on episomal plasmid uptake, and no impact on integrative plasmid transformation. Other experiments reveal the ability of some phages to evade or circumvent BREX-encoded resistance, in a manner reminiscent of what was recently established for bacteriophage escape of CRISPR-Cas adaptive immune systems (Bondy-Denomy et al, 2013), further illustrating the arms race nature of host-phage dynamics. Overall, this study provides the crucial proof of concept that BREX constitutes a novel phage resistance system, which is distinct from previously characterized viral defense mechanisms, both genetically and phenotypically. This defense system allows phage absorption and DNA integration into the host cell, but precludes viral replication in a methylation-dependent manner, unraveling a role for epigenetics in bacterial virus resistance. Though present in only 10% of microbial genomes, this innate immune system is phylogenetically widespread. Future studies should determine the molecular underpinning of BREX-encoded immunity in bacteria and shed light on the biochemical processes that drive host chromosome methylation-dependent phage resistance. Additional studies should assess whether this system has a fitness cost and whether it coexists with other defense systems, notably R-M, Abi and CRISPR. The findings presented in Goldfarb et al advance our understanding of the role of phage resistance systems in the arms race between bacterial communities and their viral predators and potentially open new avenues for engineered phage resistance in bacteria. References Barrangou R, Marraffini LA (2014) CRISPR-Cas systems: prokaryotes upgrade to adaptive immunity. Mol Cell 54: 234–244CrossrefCASPubMedWeb of Science®Google Scholar Bondy-Denomy J, Pawluk A, Maxwell KL, Davidson AR (2013) Bacteriophage genes that inactivate the CRISPR/Cas bacterial immune system. Nature 493: 429–432CrossrefCASPubMedWeb of Science®Google Scholar Chinenova TA, Mkrtumian NM, Momovskaia ND (1982) Genetic characteristics of a new phage resistance trait in Streptomyces coelicolor A3(2). Genetika 18: 1945–1952CASPubMedWeb of Science®Google Scholar Chopin MC, Chopin A, Bidnenko E (2005) Phage abortive infection in lactococci: variations on a theme. Curr Opin Microbiol 8: 473–479CrossrefCASPubMedWeb of Science®Google Scholar Goldfarb T, Sberro H, Weinstock W, Cohen O, Doron S, Charpak A, Afik S, Ofir G, Sorek R (2015) BREX, a phage resistance system widespread in microbial genomes. EMBO J 34: 169–183Wiley Online LibraryPubMedWeb of Science®Google Scholar Labrie SJ, Samson JE, Moineau S (2010) Bacteriophage resistance mechanisms. Nat Rev Microbiol 8: 317–327CrossrefCASPubMedWeb of Science®Google Scholar Makarova KS, Wolf YI, Snir S, Koonin EV (2011) Defense islands in bacterial and archaeal genomes and prediction of novel defense systems. J Bacteriol 193: 6039–6056CrossrefCASPubMedWeb of Science®Google Scholar Swarts DC, Makarova K, Wang Y, Nakanishi K, Ketting RF, Koonin EV, Patel DJ, van der Oost J (2014) The evolutionary journey of the Argonaute proteins. Nat Struct Mol Biol 21: 743–753CrossrefCASPubMedWeb of Science®Google Scholar Tock MR, Dryden DT (2005) The biology of restriction and anti-restriction. Curr Opin Microbiol 8: 466–472CrossrefCASPubMedWeb of Science®Google Scholar Van der Oost J, Westra ER, Jackson RN, Wiedenheft B (2014) Unravelling the structural and mechanistic basis of CRISPR-Cas systems. Nat Rev Microbiol 12: 479–492CrossrefCASPubMedWeb of Science®Google Scholar Previous ArticleNext Article Read MoreAbout the coverClose modalView large imageVolume 34,Issue 2,14 January 2015Cover: Samburu lioness. A lioness marches across the grasslands of Samburu in northern Kenya. The moment was captured by Susan McConnell, a developmental neurobiologist and conservation photographer from Stanford, CA. You can explore more of her stunning photography at http://www.susankmcconnell.com. Volume 34Issue 214 January 2015In this issue FiguresReferencesRelatedDetailsLoading ... DA - 2015/1/14/ PY - 2015/1/14/ DO - 10.15252/embj.201490620 VL - 34 IS - 2 SP - 134-135 SN - 1460-2075 ER - TY - JOUR TI - A Pilot Study on the Protein Composition of Induced Nonpuerperal Human Milk AU - Perrin, Maryanne Tigchelaar AU - Wilson, Erica AU - Chetwynd, Ellen AU - Fogleman, April T2 - JOURNAL OF HUMAN LACTATION AB - Our understanding of the components of human puerperal milk is extensive and increasing, yet the literature on nonpuerperal human milk has been limited to studies that measure the success of induced lactation.This study aimed to describe the composition of total protein and key bioactive proteins when lactation is induced in nonpuerperal women.Two women who induced lactation in the absence of pregnancy provided weekly milk samples over a 2-month period for analysis of total protein, secretory immunoglobulin A (sIgA), lysozyme, and lactoferrin. Composition was compared to the mature milk of 3 puerperal control subjects who were 11 months postpartum.Median total protein for subject A was 2.30 g/dL (interquartile range [IQR] = 0.46) and showed a significant downward trend over time (P < .0001), whereas the median total protein for subject B was 2.21 g/dL (IQR = 0.18) and showed a nonsignificant decline (P = .232). Total protein in both nonpuerperal subjects was elevated compared to control subjects. Secretory IgA activity declined for both nonpuerperal subjects over time, whereas lysozyme concentrations increased over time. Both sIgA and lysozyme approached the levels seen in the puerperal controls. Lactoferrin levels remained stable for both nonpuerperal subjects and were elevated compared to puerperal milk samples.This pilot study suggests that nonpuerperal milk has similar or higher levels of total protein, sIgA, lactoferrin, and lysozyme compared to puerperal, mature milk at 11 months postpartum, which warrants more attention as adoptive mothers increasingly choose to induce lactation. DA - 2015/2// PY - 2015/2// DO - 10.1177/0890334414552827 VL - 31 IS - 1 SP - 166-171 SN - 1552-5732 KW - adopted children KW - breastfeeding KW - human milk KW - milk composition ER - TY - JOUR TI - Validation of a novel rinse and filtration method for efficient processing of fresh produce samples for microbiological indicator enumeration AU - Heredia, N. AU - Solis-Soto, L. AU - Venegas, F. AU - Bartz, F. E. AU - Aceituno, A. F. AU - Jaykus, L. A. AU - Leon, J. S. AU - Garcia, S. T2 - Journal of Food Protection AB - Several methods have been described to prepare fresh produce samples for microbiological analysis, each with its own advantages and disadvantages. The aim of this study was to compare the performance of a novel combined rinse and membrane filtration method to two alternative sample preparation methods for the quantification of indicator microorganisms from fresh produce. Decontaminated cantaloupe melons and jalapeño peppers were surface inoculated with a cocktail containing 10(6) CFU/ml Escherichia coli, Salmonella Typhimurium, and Enterococcus faecalis. Samples were processed using a rinse and filtration method, homogenization by stomacher, or a sponge-rubbing method, followed by quantification of bacterial load using culture methods. Recovery efficiencies of the three methods were compared. On inoculated cantaloupes, the rinse and filtration method had higher recovery of coliforms (0.95 log CFU/ml higher recovery, P = 0.0193) than the sponge-rubbing method. Similarly, on inoculated jalapeños, the rinse and filtration method had higher recovery for coliforms (0.84 log CFU/ml higher, P = 0.0130) and E. coli (1.46 log CFU/ml higher, P < 0.0001) than the sponge-rubbing method. For jalapeños, the rinse and filtration method outperformed the homogenization method for all three indicators (0.79 to 1.71 log CFU/ml higher, P values ranging from 0.0075 to 0.0002). The precision of the three methods was also compared. The precision of the rinse and filtration method was similar to that of the other methods for recovery of two of three indicators from cantaloupe (E. coli P = 0.7685, E. faecalis P = 0.1545) and was more precise for recovery of two of three indicators from jalapeño (coliforms P = 0.0026, E. coli P = 0.0243). Overall, the rinse and filtration method performed equivalent to, and sometimes better than, either of the compared methods. The rinse and filtration method may have logistical advantages when processing large numbers of samples, improving sampling efficiency and facilitating microbial detection. DA - 2015/// PY - 2015/// DO - 10.4315/0362-028x.jfp-14-324 VL - 78 IS - 3 SP - 525-530 ER - TY - JOUR TI - Formation of soluble whey protein aggregates and their stability in beverages AU - Ryan, Kelsey N. AU - Foegeding, E. Allen T2 - FOOD HYDROCOLLOIDS AB - The objectives of this work were to 1) determine the role of pH in forming thermally stable whey protein isolate (WPI) soluble aggregates (SA), 2) evaluate the functionality of SA formed under a range of conditions, and 3) evaluate SA stability in a model beverage. Whey protein isolate SA were formed using two sources of WPI at pH 6.5 or 7.5, and thermal stability was tested in the presence of 30–60 mM NaCl or 15–30 mM CaCl2. Soluble aggregates from the two sources of WPI exhibited physicochemical and functional differences over the range of conditions investigated. Soluble aggregates formed at pH 7.5, 7% w/w protein, by heating statically at 90 °C for 10 min exhibited lower turbidity and viscosity, and higher solubility when thermally treated in solutions containing salt for both types of WPI, and were more sensitive to CaCl2. Soluble aggregates and a model beverage containing the SA were prepared at pilot scale and evaluated for shelf stability. Soluble aggregates were formed under continuous flow at pH 6.5, 7% w/w WPI, added at 5% w/w to the beverage, and thermally processed. Over nine weeks of storage at 22.5 °C, the beverage with SA had lower turbidity, aggregate size, and viscosity; and the aggregates had a more negative zeta potential. Results indicate that SA can be used to improve thermal and storage stability in beverages containing salts, and that solution and heating conditions can be optimized for improved thermal stability. DA - 2015/1// PY - 2015/1// DO - 10.1016/j.foodhyd.2014.05.025 VL - 43 SP - 265-274 SN - 1873-7137 KW - Protein KW - Aggregation KW - Heat stability KW - Beverages ER - TY - JOUR TI - Compounds leached from quinoa seeds inhibit matrix metalloproteinase activity and intracellular reactive oxygen species AU - Graf, B. L. AU - Cheng, D. M. AU - Esposito, D. AU - Shertel, T. AU - Poulev, A. AU - Plundrich, N. AU - Itenberg, D. AU - Dayan, N. AU - Lila, M. A. AU - Raskin, I. T2 - INTERNATIONAL JOURNAL OF COSMETIC SCIENCE AB - Synopsis Objective Quinoa ( C henopodium quinoa W illd.) is a seed crop rich in bioactive compounds including phytoecdysones (especially 20‐hydroxyecdysone, 20 HE ), polyphenols, proteins and essential fatty acids. We previously reported a method to leach and concentrate quinoa bioactives into a complex phytochemical mixture termed quinoa leachate ( QL ). Here, we aimed to determine the effect of QL and its chemically distinct fractions on five biochemical endpoints relevant to skin care applications: (i) cell viability, (ii) matrix metalloproteinase ( MMP ) m RNA expression, (iii) MMP enzymatic activity, (iv) tyrosinase enzymatic activity and (v) intracellular reactive oxygen species ( ROS ) production. Methods Quinoa leachate was fractionated and chemically characterized using column chromatography and liquid chromatography–mass spectrometry ( LC ‐ MS ). Cell viability was determined using a MTT assay in four mammalian cell lines. MMP ‐1 m RNA expression was assessed in human dermal fibroblasts ( HDF ) via q RT ‐ PCR . The enzymatic activity of MMP ‐9 and tyrosinase was measured using fluorometric and colorimetric in vitro assays, respectively. Lipopolysaccharide ( LPS )‐induced ROS production was determined in human dermal fibroblasts by fluorescence intensity of an oxidant‐sensitive probe. Results Quinoa leachate was separated into three fractions: (i) carbohydrate‐rich fraction ( QL ‐ C ; 71.3% w/w of QL ); (ii) phytoecdysone, polyphenol and protein‐rich fraction ( QL ‐ P , 13.3% w/w of QL ); (iii) oil‐rich fraction ( QL ‐ O , 10.8% w/w of QL ). QL did not reduce cell viability in any of the four cell lines tested. QL , QL ‐ P and QL ‐ O each significantly inhibited MMP ‐1 m RNA expression in HDF at a concentration of 5 μg mL −1 . QL and QL ‐ P also significantly inhibited MMP ‐9 enzymatic activity, whereas QL ‐ P demonstrated significant tyrosinase enzymatic inhibition. Furthermore, QL , QL ‐ P , QL ‐ O and 20 HE significantly inhibited intracellular ROS production. Conclusion This study is the first to demonstrate the MMP , tyrosinase and ROS inhibiting properties of multiple different phytochemical components derived from quinoa seeds. Our work indicates that quinoa phytochemicals may play a role in the treatment and prevention of skin ageing through a multiplicity of effects. DA - 2015/4// PY - 2015/4// DO - 10.1111/ics.12185 VL - 37 IS - 2 SP - 212-221 SN - 1468-2494 KW - ageing KW - cell culture KW - chemical analysis KW - oxidative damage KW - phytochemistry KW - phytoecdysteroid ER - TY - JOUR TI - Cleaning and sanitation of Salmonella-contaminated peanut butter processing equipment AU - Grasso, Elizabeth M. AU - Grove, Stephen F. AU - Halik, Lindsay A. AU - Arritt, Fletcher AU - Keller, Susanne E. T2 - FOOD MICROBIOLOGY AB - Microbial contamination of peanut butter by Salmonella poses a significant health risk as Salmonella may remain viable throughout the product shelf life. Effective cleaning and sanitation of processing lines are essential for preventing cross-contamination. The objective of this study was to evaluate the efficacy of a cleaning and sanitation procedure involving hot oil and 60% isopropanol, ± quaternary ammonium compounds, to decontaminate pilot-scale processing equipment harboring Salmonella. Peanut butter inoculated with a cocktail of four Salmonella serovars (∼7 log CFU/g) was used to contaminate the equipment (∼75 L). The system was then emptied of peanut butter and treated with hot oil (90 °C) for 2 h followed by sanitizer for 1 h. Microbial analysis of food-contact surfaces (7 locations), peanut butter, and oil were conducted. Oil contained ∼3.2 log CFU/mL on both trypticase soy agar with yeast extract (TSAYE) and xylose lysine deoxycholate (XLD), indicating hot oil alone was not sufficient to inactivate Salmonella. Environmental sampling found 0.25–1.12 log CFU/cm2 remaining on processing equipment. After the isopropanol sanitation (±quaternary ammonium compounds), no Salmonella was detected in environmental samples on XLD (<0.16 log CFU/cm2). These data suggest that a two-step hot oil clean and isopropanol sanitization treatment may eliminate pathogenic Salmonella from contaminated equipment. DA - 2015/4// PY - 2015/4// DO - 10.1016/j.fm.2014.03.003 VL - 46 SP - 100-106 SN - 1095-9998 KW - Peanut butter KW - Salmonella KW - Sanitation ER - TY - JOUR TI - Sugar reduction of skim chocolate milk and viability of alternative sweetening through lactose hydrolysis AU - Li, X. E. AU - Lopetcharat, K. AU - Qiu, Y. AU - Drake, M. A. T2 - JOURNAL OF DAIRY SCIENCE AB - Milk consumption by Americans has not met the standards of the Dietary Guidelines for Americans. Chocolate milk can improve milk consumption, especially by children, due to its color and taste. However, the high sugar content of chocolate milk is a cause for concern about its healthfulness, resulting in its removal from some school lunch programs. It is important to reduce the sugar content of chocolate milk and still maintain acceptability among consumers. It is also important to investigate other natural alternatives to sweetening. The objectives of this study were to identify the different sweetness intensity perceptions of sucrose in water and various dairy matrices, to identify the acceptable reduction in sweet taste for chocolate milk for both young adults (19–35 yr) and children (5–13 yr), and to determine if lactose hydrolysis is a viable alternative. Threshold and power function studies were used to determine the benchmark concentration of sucrose in chocolate milk. The acceptability of sugar reduction from the benchmark concentration for both young adults and children and the acceptability of lactose hydrolyzed chocolate milk (4°C for 24 h) with added lactose for young adults were evaluated. Acceptability results demonstrated that sugar reduction in chocolate milk is possible for both young adults and children as long as it does not exceed a 30% reduction (from 205 mM). Lactose hydrolysis of added lactose was used to achieve the sweetness of sucrose in chocolate milk but required >7.5% (wt/vol) added lactose, which contributed undesirable calories, indicating that lactose hydrolysis may be more suitable for other dairy beverages that require less added sugar. The findings of this study demonstrate consumer acceptance of reduced-sugar chocolate milk and a possible way to use lactose hydrolysis in dairy beverages. DA - 2015/3// PY - 2015/3// DO - 10.3168/jds.2014-8490 VL - 98 IS - 3 SP - 1455-1466 SN - 1525-3198 KW - chocolate milk KW - power function KW - sugar reduction KW - lactose hydrolysis ER - TY - JOUR TI - Short communication: The influence of solids concentration and bleaching agent on bleaching efficacy and flavor of sweet whey powder AU - Jervis, M. G. AU - Smith, T. J. AU - Drake, M. A. T2 - JOURNAL OF DAIRY SCIENCE AB - Recent studies have demonstrated the effect of bleaching conditions and bleaching agent on flavor and functional properties of whey protein ingredients. Solids concentration at bleaching significantly affected bleaching efficacy and flavor effects of different bleaching agents. It is not known if these parameters influence quality of sweet whey powder (SWP). The purpose of this study was to determine the effects of solids concentration and bleaching agent on the flavor and bleaching efficacy of SWP. Colored cheddar whey was manufactured, fat separated, and pasteurized. Subsequently, the whey (6.7% solids) was bleached, concentrated using reverse osmosis (RO) to 14% solids, and then spray dried, or whey was concentrated before bleaching and then spray dried. Bleaching treatments included a control (no bleaching, 50°C, 60 min), hydrogen peroxide (HP; 250 mg/kg, 50°C, 60 min), benzoyl peroxide (50 mg/kg, 50°C, 60 min), lactoperoxidase (20 mg/kg of HP, 50°C, 30 min), and external peroxidase (MaxiBright, DSM Food Specialties, Delft, the Netherlands; 2 dairy bleaching units/mL, 50°C, 30 min). The experiment was repeated in triplicate. Sensory properties and volatile compounds of SWP were evaluated by a trained panel and gas chromatography-mass spectrometry, respectively. Bleaching efficacy (norbixin destruction) and benzoic acid were measured by HPLC. Differences in bleaching efficacy, sensory and volatile compound profiles, and benzoic acid were observed with different bleaching agents, consistent with previous studies. Solids concentration affected bleaching efficacy of HP, but not other bleaching agents. The SWP from whey bleached with HP or lactoperoxidase following RO had increased cardboard and fatty flavors and higher concentrations of lipid oxidation compounds compared with SWP from whey bleached before RO. The SWP bleached with benzoyl peroxide after RO contained less benzoic acid than SWP from whey bleached before RO. These results indicate that solids concentration at bleaching and bleaching agent affect quality of SWP. DA - 2015/4// PY - 2015/4// DO - 10.3168/jds.2014-8804 VL - 98 IS - 4 SP - 2294-2302 SN - 1525-3198 KW - sweet whey KW - reverse osmosis KW - bleaching ER - TY - JOUR TI - Persistence of Human Norovirus RT-qPCR Signals in Simulated Gastric Fluid AU - Tung-Thompson, Grace AU - Gentry-Shields, Jennifer AU - Fraser, Angela AU - Jaykus, Lee-Ann T2 - FOOD AND ENVIRONMENTAL VIROLOGY DA - 2015/3// PY - 2015/3// DO - 10.1007/s12560-014-9170-4 VL - 7 IS - 1 SP - 32-40 SN - 1867-0342 KW - Norovirus KW - Persistence KW - Simulated gastric fluid KW - Vomitus ER - TY - JOUR TI - Investigating the filled gel model in Cheddar cheese through use of Sephadex beads AU - Barden, L. M. AU - Osborne, J. A. AU - McMahon, D. J. AU - Foegeding, E. A. T2 - JOURNAL OF DAIRY SCIENCE AB - Cheese can be modeled as a filled gel whereby milkfat globules are dispersed in a casein gel network. We determined the filler effects using Sephadex beads (GE Healthcare Life Sciences, Pittsburgh, PA) as a model filler particle. Ideally, such a model could be used to test novel filler particles to replace milkfat in low-fat cheese. Low-filler (6% particles), reduced-filler (16%), and full-filler (33%) cheeses were produced using either Sephadex beads of varying sizes (20 to 150 μm diameter) or milkfat. Small- and large-strain rheological tests were run on each treatment at 8, 12, and 18 wk after cheese manufacturing. Differences in rheological properties were caused primarily by the main effects of filler volume and type (milkfat vs. Sephadex), whereas filler size had no obvious effect. All treatments showed a decrease in deformability and an increase in firmness as filler volume increased above 25%, although the beads exhibited a greater reinforcing effect and greater energy recovery than milkfat. DA - 2015/3// PY - 2015/3// DO - 10.3168/jds.2014-8597 VL - 98 IS - 3 SP - 1502-1516 SN - 1525-3198 KW - cheese KW - filled gel KW - Sephadex KW - particle size ER - TY - JOUR TI - Effect of serum cotinine on vitamin D serum concentrations among American females with different ethnic backgrounds AU - Manavi, K. R. AU - Alston-Mills, B. P. AU - Thompson, M. P. AU - Allen, J. C. T2 - Anticancer Research DA - 2015/// PY - 2015/// VL - 35 IS - 2 SP - 1211-1218 ER - TY - JOUR TI - Feeding practices of low-income mothers: how do they compare to current recommendations? AU - Power, Thomas G. AU - Hughes, Sheryl O. AU - Goodell, L. Suzanne AU - Johnson, Susan L. AU - Duran, J. Andrea Jaramillo AU - Williams, Kimberly AU - Beck, Ashley D. AU - Frankel, Leslie A. T2 - INTERNATIONAL JOURNAL OF BEHAVIORAL NUTRITION AND PHYSICAL ACTIVITY AB - Despite a growing consensus on the feeding practices associated with healthy eating patterns, few observational studies of maternal feeding practices with young children have been conducted, especially in low-income populations. The aim of this study was to provide such data on a low income sample to determine the degree to which observed maternal feeding practices compare with current recommendations.Eighty low-income mothers and their preschool children were videotaped at dinner in their homes. Mothers were chosen from a larger study to create a 2 X 2 X 2 design: maternal ethnicity (African American vs. Latina) by child gender by child weight status (healthy weight vs. overweight/obese). Observers coded videotapes for a range of maternal feeding strategies and other behaviors.Many mothers spent considerable time encouraging eating--often in spite of the child's insistence that he or she was finished. Mothers talked little about food characteristics, rarely referred to feelings of hunger and fullness, and made more attempts to enforce table manners than to teach eating skills. Latina mothers showed higher levels of teaching eating skills and encouraging their children to eat; African American mothers showed higher levels of enforcing table manners and getting children to clear their plates. Mothers of boys used more unelaborated commands and less questions/suggestions than mothers of girls. Finally, compared to mothers of overweight/obese children, mothers of healthy weight children showed higher levels of encouraging eating and lower levels of discouraging eating.Most of the mothers in this study did not engage in feeding practices that are consistent with current recommendations. They did this, despite the fact that they knew they were being observed. These results should be used to inform future research about the motivations behind mothers' feeding practices and the development of interventions by helping identify areas in greatest need of change. DA - 2015/3/7/ PY - 2015/3/7/ DO - 10.1186/s12966-015-0179-3 VL - 12 SP - SN - 1479-5868 KW - Mothers KW - Feeding behaviors KW - Obesity ER - TY - JOUR TI - An observational study of frequency of provider hand contacts in child care facilities in North Carolina and South Carolina AU - Fraser, Angela AU - Wohlgenant, Kelly AU - Cates, Sheryl AU - Chen, , Xi AU - Jaykus, Lee-Ann AU - Li, You AU - Chapman, Benjamin T2 - AMERICAN JOURNAL OF INFECTION CONTROL AB - •Children's clothing are the most frequently touched. •Common high-touch surfaces, such as light switches and doorknobs, are touched the least. •Guidelines are available for nonporous surfaces but not for porous surfaces. Background Children enrolled in child care are 2.3-3.5 times more likely to experience acute gastrointestinal illness than children cared for in their own homes. The purpose of this study was to determine the frequency surfaces were touched by child care providers to identify surfaces that should be cleaned and sanitized. Methods Observation data from a convenience sample of 37 child care facilities in North Carolina and South Carolina were analyzed. Trained data collectors used iPods (Apple, Cupertino, CA) to record hand touch events of 1 child care provider for 45 minutes in up to 2 classrooms in each facility. Results Across the 37 facilities, 10,134 hand contacts were observed in 51 classrooms. Most (4,536) were contacts with porous surfaces, with an average of 88.9 events per classroom observation. The most frequently touched porous surface was children's clothing. The most frequently touched nonporous surface was food contact surfaces (18.6 contacts/observation). Surfaces commonly identified as high-touch surfaces (ie, light switches, handrails, doorknobs) were touched the least. Conclusion General cleaning and sanitizing guidelines should include detailed procedures for cleaning and sanitizing high-touch surfaces (ie, clothes, furniture, soft toys). Guidelines are available for nonporous surfaces but not for porous surfaces (eg, clothing, carpeting). Additional research is needed to inform the development of evidence-based practices to effectively treat porous surfaces. Children enrolled in child care are 2.3-3.5 times more likely to experience acute gastrointestinal illness than children cared for in their own homes. The purpose of this study was to determine the frequency surfaces were touched by child care providers to identify surfaces that should be cleaned and sanitized. Observation data from a convenience sample of 37 child care facilities in North Carolina and South Carolina were analyzed. Trained data collectors used iPods (Apple, Cupertino, CA) to record hand touch events of 1 child care provider for 45 minutes in up to 2 classrooms in each facility. Across the 37 facilities, 10,134 hand contacts were observed in 51 classrooms. Most (4,536) were contacts with porous surfaces, with an average of 88.9 events per classroom observation. The most frequently touched porous surface was children's clothing. The most frequently touched nonporous surface was food contact surfaces (18.6 contacts/observation). Surfaces commonly identified as high-touch surfaces (ie, light switches, handrails, doorknobs) were touched the least. General cleaning and sanitizing guidelines should include detailed procedures for cleaning and sanitizing high-touch surfaces (ie, clothes, furniture, soft toys). Guidelines are available for nonporous surfaces but not for porous surfaces (eg, clothing, carpeting). Additional research is needed to inform the development of evidence-based practices to effectively treat porous surfaces. DA - 2015/2/1/ PY - 2015/2/1/ DO - 10.1016/j.ajic.2014.10.017 VL - 43 IS - 2 SP - 107-111 SN - 1527-3296 KW - Child care providers KW - Porous surfaces KW - Nonporous surfaces KW - Hand contact KW - Observational study ER - TY - JOUR TI - High pressure effects on heat-induced gelation of threadfin bream (Nemipterus spp.) surimi AU - Zhu, Zhiwei AU - Lanier, Tyre C. AU - Farkas, Brian E. T2 - JOURNAL OF FOOD ENGINEERING AB - We sought to determine, in threadfin bream surimi system, if 40 °C incubation carried out simultaneous with, or following, HPP might also induce a more effective setting on subsequently cooked gels. Threadfin bream surimi pastes were subjected to HPP 200 or 300; at 5 °C (little or no setting effect expected during HPP at this temperature) for 15 min or at 40 °C (optimal setting temperature for threadfin bream pastes) for 15 or 30 min. Pastes were then directly cooked (90 °C for 20 min) (pressureP°C/time > C), or first allowed to undergo setting at atmospheric pressure (40 °C for 60 min), followed by cooking (90 °C 20 min) (pressureP°C/time > Stime > C). With or without followed setting treatment at 40 °C, HPP treatment at 40 °C, or even lower pressure at 5 °C for shorter time (200MPaP5°C/15min > S40°C/60min > C) produce weak cooking gels. Two opposite actions of cross-linked polymer (CP) and degraded protein (DP) was observed during the HPP treatment. The given HPP treatment was sure to triggered the detriment of protein in combined treatments, the followed setting treatment after HPP enhanced the degradation of protein as well. The formation of disulfide bonds during HPP and cooking treatments was retarded, which was attributed to the influence of protein degradation behavior. Scanning electron microscopy revealed, with giving a prior HPP treatment, the fiberlike structure gradually disappeared and the compact microstructure increased in final cooking gel. DA - 2015/2// PY - 2015/2// DO - 10.1016/j.jfoodeng.2014.08.021 VL - 146 SP - 23-27 SN - 1873-5770 KW - High pressure KW - Setting KW - Heat-induced gelation KW - Threadfin bream KW - Surimi ER -