bluuebunny/crossref_metadata_2025 huggingface dataset:
A Novel laccase from Lentinus sp.
Authors: Hsu Chih‐An , Wen Tuan‐Nan , Shyur Lie‐Fen
Published: June 2021

Laccase is known to play an important role in various industrial applications, such as biopulping, pretreatment of lignocellulosics for bioethanol production, dye decoloration, and bioremediation. A native laccase protein of Lentinus sp. was purified and identified as N‐linkage glycosylated by LC/MS‐MS analysis. Kinetics study showed that the specific activity of the purified laccase was 2047 U mg−1 at pH 2.5 and 70°C with ABTS as the substrate. The kcat, Km and catalytic efficiency (kcat/Km) of the enzyme were 2017 s−1, 8.4 μM, and 240 s−1μM−1, respectively. The enzyme was stable at pH 2.5–6.0, and t1/2 of enzymatic activity was approximately of 20 min at 70°C and pH 2.5 and 118 h at 25ºC and pH 6.0. Differential scanning calorimetry analysis demonstrated that the laccase possesses a high Tm value at 77.1°C. Glycosylation was found to play an important role to the enzymatic activity. N75D, N236D, N399D, and N458D mutant laccases exhibited approximately 80–100% decreases in enzyme activity. The laccase showed a good stability against ethanol and remained 80% of the original activity after a 120‐h incubation in 25% ethanol. We found 34% lignin degradation efficiency on rice straw with crude laccase enzymes, and 47% decoloration efficiency on Remazol Brilliant Blue R with partial purified laccase. The fungal laccase identified in this study possesses novel functions with great potential in bio‐industrial applications.
Characterization by Mutagenesis of Heme Binding Pocket Interactions in Caenorhabditis elegans Dual Oxidase 1 (DUOX1)
Authors: Meitzler Jennifer L. , Ortiz de Montellano Paul R.
Published: June 2021

The DUOX members of the NOX protein family, which are found throughout eukaryotic species, including Caenorhabditis elegans and sea urchins, have garnered much attention for their unique N‐terminal peroxidase‐like domain(s) and their ability to produce hydrogen peroxide. Two highly conserved DUOX isoforms are encoded in the C. elegans genome; CeDUOX1/BLI‐3 and CeDUOX2. RNAi experiments in C. elegans to knock out the expression of nematode DUOX1 have found that this protein participates in the biogenesis of the nematode cuticle through formation of di‐ and trityrosine linkages. Peroxidases, including mammalian peroxidases and ovoperoxidases, have been identified as enzymes capable of catalyzing such cross‐linking in the presence of hydrogen peroxide. However, the absence of key active site residues within the N‐terminal domain of DUOX(s) makes their heme binding and catalytic activity uncertain. To further characterize the function and mechanism of action of the DUOX peroxidase‐like domain, the N‐terminal region of C. elegans DUOX1 was expressed and purified from a baculovirus system. Evaluation of this protein demonstrated that it binds heme covalently and exhibits a modest peroxidase activity. Mutation studies of this protein conducted to investigate the role of residues interacting with the co‐factor in the heme binding pocket will be presented. This work was partially supported by NIH Grant GM32488.
Requirement of conserved amino acid residues for ER‐export and recycling of platelet‐activating factor receptor
Authors: Nakamura Motonao , Hirota Nobuaki , Yasuda Daisuke , Hashidate Tomomi , Yamaguchi Satoshi , Nagamune Teruyuki , Shimizu Takao
Published: June 2021

In the transmembrane domains (TMs) of G‐protein coupled receptors, several amino acid residues are conserved. Here, we demonstrate that a conserved proline, Pro247, in TM6 of platelet‐activating factor receptor (PAFR) is critical for endoplasmic reticulum (ER)‐export and recycling after agonist‐induced internalization. Alanine‐substituted mutants of the conserved residues of PAFRs, including P247A, were accumulated in the ER. Because a PAFR antagonist, Y‐24180, acted as a pharmacological chaperone to rescue ER‐retention, this retention is due to misfolding of PAFR. mc‐PAF, a PAFR agonist, did not increase surface expression of P247A, even if another ER‐retained mutant, D63A, was trafficked. Signaling and accumulation of the receptors in the early endosomes were observed in the mc‐PAF‐treated P247A‐expressing cells, suggesting that P247A was trafficked to the cell surface by mc‐PAF, thereafter internalized, but not recycled. Deficient recycling was confirmed with a sortase‐A‐mediated method for labeling cell surface proteins. These results demonstrate that the conserved proline in TM6 is crucial for the intracellular trafficking of PAFR. Supported in part by a Grants‐in‐aid from Ministry of Education, Culture, Sports, Science, and Technology of Japan
PROTEOME DYNAMICS ASSOCIATED TO THE PATHOBIOLOGY OF ALZHEIMER'S DISEASE
Authors: De Jesús Héctor José
Published: June 2021

The central role of the microtubule associated protein tau in neurodegeneration is underscored by the identification of mutations on the tau gene in patients with frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP‐17), corticobasal degeneration (CBD), and progressive supranuclear palsy (PSP), as well as been considered as a pathological hallmark in Alzheimer's disease. Several hypotheses have been proposed to explain the pathobiological mechanisms induced in neurodegenerative diseases associated to tau pathology, collectively known as tauopathies. However, the molecular mechanisms underlying tau‐mediated neurodegeneration are poorly understood. The main goal of our studies is to identify and characterize proteome changes underlying tau‐mediated neurodegeneration. Proteome dynamics could be defined as transient changes in post‐translational modification, interactions, function and/or subcellular localization. Taking this concept as a research approach has led us to the identification of protein level changes associated to tau‐mediated neurodegeneration. The identification of affected proteins may provide insights on the pathobiological process linked to tau‐mediated neurodegeneration. The approach, result and significance of the findings will be discussed.
A structural model for catalysis by tyrosyl‐tRNA synthetase
Authors: First Eric Allen
Published: June 2021

Twenty‐eight tyrosyl‐tRNA synthetase (TyrRS) structures were aligned and their active sites were compared. This analysis revealed that the HIGH signature sequence adopts two distinct conformations that correlate with the open and closed forms of the KMSKS loop. Based on this analysis, we propose a model in which the binding of ATP forces HIGH to adopt a closed conformation. This forces the KMSKS loop to adopt a closed, strained, conformation, bringing the KMSKS sequence into the active site and positioning it to interact with pyrophosphate in the TyrRS·[Tyr‐ATP]‡ transition state. Formation of the transition state results in cleavage of the scissile bond in ATP, releasing the KMSKS loop from the closed (strained) conformation. This mechanism provides a structural explanation for the observations that: (1) tyrosyl‐tRNA synthetase uses ATP‐binding energy to stabilize the transition state of the reaction and (2) deletion of the KMSKS sequence increases the binding affinity of the TyrRS·Tyr complex for ATP. Lastly, structural comparison of eukaryotic and archaeal tyrosyl‐tRNA synthetases revealed a ligand‐dependent conformational change that links the tyrosine binding site to the dimer interface, suggesting a possible model for the observation that tyrosyl‐tRNA synthetase displays half‐of‐the‐sites reactivity with respect to the binding of tyrosine.

This research was funded NIH GM68070 and MDA 4268.
LC‐MS/MS determination of phosphatidylcholine hydroperoxide and its truncated products in human blood plasma (1001.9)
Authors: Miyazawa Teruo , Kato Shunji , Sawane Kento , Miyazawa Taiki , Nakagawa Kiyotaka
Published: June 2021

Accumulation of phosphatidylcholine hydroperoxide (PCOOH), a primary oxidation product of phosphatidylcholine (PC), in the blood plasma has been observed in vascular pathological conditions, including inflammation and atherosclerosis. Here, we developed a method for quantification of PCOOH (1‐palmitoyl‐2‐hydroperoxyoctadecadienoyl‐sn‐glycero‐3‐phosphocholine, 16:0/HODE PC), especially isomers such as 16:0/13‐HODE PC and 16:0/9‐HODE PC that might be present in human plasma, together with their breakdown products (truncated PC such as 16:0/azelaoyl PC), by using LC‐MS/MS. Collision‐induced dissociation of sodiated PCOOH isomers ([M+Na]+, m/z 812) produced not only a fragment ion (m/z 147) but also characteristic neutral losses (88 Da for 16:0/13‐HODE PC and 169 Da for 16:0/9‐HODE PC). Three multiple reaction monitorings (MRMs) of sodiated PCOOH could be performed under optimum conditions. MRM (812/147) enabled determination of 16:0/HODE PC, and MRM (812/541 and 812/388) allowed measurement of 16:0/13‐HODE PC and 16:0/9‐HODE PC, respectively. By using the method, we could determine plasma PCOOH concentrations in healthy subjects (260nmol/L of total PCOOH, 50‐80nmol/L of truncated PC, 36nmol/L of 16:0/13‐HODE PC and 33nmol/L of 16:0/9‐HODE PC) and in patients with angiographically significant stenosis (45nmol/L of 16:0/13‐HODE PC, 51 nmol/L of 16:0/9‐HODE PC). The PCOOH detected is formed with lipoxygenase and free radical reaction. Oxidized PC with singlet molecular oxygen was not found. The newly developed LC‐MS/MS method appears to be a powerful tool for understanding in vivo membrane lipid peroxidation.
Chemoenzymatic synthesis of HIV‐1 V1V2 glycopeptide antigens for epitope characterization and neutralizing antibody detection (1004.4)
Authors: Amin Mohammed , Lomino Joseph , Wang Lai‐Xi
Published: June 2021

Recent discovery of a new class of glycan‐dependent broadly neutralizing antibodies (bnAbs), including PG9, PG16, and the PGT series antibodies, provides valuable templates for HIV‐1 vaccine design. Epitope mapping suggests that PG9 and PG16 recognize unique glycopeptide antigens located in the V1V2 region of gp120. However, further characterization of the neutralizing epitopes has been difficult due to the glycosylation heterogeneity of gp120. In this paper, we report the design, synthesis, and antigenicity of well‐defined V1V2 glycopeptides aiming to characterize the glycan specificity of the epitopes. Homogeneous V1V2 glycopeptides carrying defined N‐glycans at the glycosylation sites (N160 and N156 or N173) were synthesized by a convergent chemoenzymatic approach. Binding studies confirmed the essence of a Man5GlcNAc2 glycan at Asn160 for recognition by PG9 and PG16 and further revealed a critical role of a sialylated N‐glycan at the secondary site (N156 or N173) in the context of glycopeptides. We have also synthesized novel V1V2 glycopeptides in which the innermost GlcNAc was replaced with a Glc moiety. Binding studies show that the Glc replacement did not impair antibody recognition. The usefulness of the synthetic V1V2 glycopeptides as coating antigens for neutralizing antibody detection was demonstrated in an ELISA format, which enables the detection of PG9 and PG16 at up to 50 pM concentration. The high‐affinity synthetic glycopeptides identified also provides a basis for HIV‐1 vaccine design.

Grant Funding Source: Supported by NIH Grant 1R21AI101035 to L‐XW
Expression of the metacaspase cp3 from the fungus Schizophyllum commune in Escherichia coli (1010.9)
Authors: Spencer Zachary , Gagliardi Connor , Fox Kristin
Published: June 2021

Programmed cell death (PCD) is a cellular suicide protocol that eliminates eukaryotic cells in a highly controlled manner. Traditionally, proteins known as caspases have been the main focus of study concerning PCD. Only recently has a related set of PCD regulatory and execution proteins been identified in non‐metazoans that lack caspases; these proteins are called metacaspases. A set of five putative metacaspases native to the fungus Schizophyllum commune were identified using a PSI‐BLAST search, and named Scp1‐5. A very strong indicator of similar function between the metacaspase and caspase protease families is the presence of a highly conserved Cys‐His catalytic dyad found throughout the known metacaspase family. In order to further characterize these proteases, Scp3’s gene sequence was cloned into two different expression plasmids that utilize two separate expression pathways. The first, pQE‐80L (Addgene), uses a phage T5 promoter to recruit native E.coli RNA polymerases. The second plasmid pET His6 Sumo TEV LIC cloning vector (Addgene), uses the T7 expression method. The latter is currently being used in order to test the efficacy of the small ubiquitin‐related modifier (SUMO) in solubilizing the overexpressed Scp3 protein that resides in inclusion bodies when the pQE‐80L system is used.
Analysis of the expression of eNOS in erythrocytes under hypoxic conditions (1013.15)
Authors: Nava Pérez Tania , Estrada José , Contreras Irazú , Moreno‐Altamirano Bertha , Pliego‐Rivero Francisco
Published: June 2021

Background: Congenital heart diseases are among the most common birth defects, being the second leading cause of infant death in Mexico. Some of these pathologies are characterized by low concentrations of O2 in the blood, resulting in cyanosis. There is evidence that erythrocytes express active and functional eNOS, which catalyzes the production of nitric oxide (NO). NO activity in erythrocytes appears to include the regulation of membrane deformability and inhibition of platelet activation; however, the activities of eNOS, NO export to the vasculature and the relative contribution of erythrocytes to the active NO pool in circulation, have not yet been explored in detail.

Objective: To identify differences in eNOS expression and activity in erythrocytes from children with congenital heart disease and cyanosis, compared to healthy controls.

Methods: Analysis of eNOS expression and activity by western blot and fluorometry, in erythrocytes of patients diagnosed with congenital heart disease and blood O2 saturation below 80%, at the Children's Hospital‐IMIEM, Toluca, Mexico.

Results: 30 samples from patients with congenital heart disease and blood O2 saturation less than 80%, and 6 samples from healthy controls matched for age and sex, have been collected. Erythrocytes of all samples have been isolated and changes in eNOS expression are being analyzed. Enzymatic activity is being analyzed by subjecting the erythrocytes to mechanical stress and by measuring changes in intracellular calcium concentrations and NO production.
p54nrb/NONO regulates cAMP signaling and cortisol production by modulating phosphodiesterase splicing (1013.21)
Authors: Yang Jia , Sewer Marion
Published: June 2021

Phosphodiesterases (PDEs) play an integral role in modulating cellular signaling by degrading the second messenger cAMP. Mammalian PDEs are encoded by 21 different genes and are subdivided into 11 families (PDE1‐11), with more than 80 alternative splice variants generated from multiple promoters. We have shown previously that p54nrb/NONO is required for glucocorticoid biosynthesis, however the molecular mechanism by which p54nrb/NONO confers optimal cortisol production was unclear. We find that silencing p54nrb/NONO in H295R human adrenocortical cells abrogates adrenocorticotrophin (ACTH)‐stimulated cAMP production. Interestingly, the expression of multiple PDE isoforms, including PDE2, PDE3, PDE4, and PDE11 were induced in p54nrb/NONO knockdown cells. Analysis of PDE splice variants revealed that p54nrb/NONO plays a critical role in regulating PDE splicing. Additionally, whole transcriptome analysis identified that silencing p54nrb/NONO altered the splicing of multiple genes in adrenocortical cells, including cAMP responsive element binding protein 5 (CREB5), and histone deacetylase 9 (HDAC9). Collectively, our findings demonstrate that p54nrb/NONO plays a key role in regulating the capacity for cAMP signaling by controlling PDE splicing and expression, ultimately influencing glucocorticoid biosynthesis.

Grant Funding Source: Supported by NIH GM073241.
Structural and functional analyses confirm the presence of functional P‐loops and intrinsic GTPase activity in Evc protein (1013.9)
Authors: Odunuga Odutayo
Published: June 2021

Ellis van Creveld (EvC) syndrome is a rare multi‐symptom human genetic disorder clinically characterized by mild short stature, postaxial polydactyly, dysplastic nail and teeth. Evc is one of the novel proteins that have been implicated in the pathogenesis of EvC syndrome and its milder form, Weyers acrofacial dysostosis (WAD). Mutations in EVC and EVC2 genes that cause these syndromes introduce premature termination codons and frameshifts, resulting in production of defective Evc and Evc2 proteins respectively. Genetic and biochemical experiments have identified Evc and Evc2 as novel proteins that are essential for positive regulation of both Indian and sonic hedgehog (Hh) signaling pathways during bone and heart development, and pathogenesis of EvC syndrome. Two separate research reports demonstrated that Evc‐Evc2 complex interacts with smoothened (smo) at the base of the cilia to transduce Hh‐induced signal to downstream proteins. We present spectroscopic and structural evidence to show that the putative P‐loop motif in Evc is functional and that it confers GTPase activity on the protein. The implications of our findings on the mechanism of transduction and regulation of Hh‐induced signals will be discussed.
Household food insecurity is associated with childhood stunting in vulnerable populations in Lebanon (1014.3)
Authors: Ghattas Hala , Sahyoun Nadine , Sassine AnnieBelle , Barbour Jessica , Seyfert Karin , Hwalla Nahla , Nord Mark
Published: June 2021

Household food insecurity (HFI) has been shown to be an independent predictor of poor child nutritional status in various settings. In the Middle East region, no data exist on the effect of directly measured HFI on children’s nutritional outcomes. Using data from three household surveys of at‐risk communities in Lebanon, we examined the effect of HFI on nutritional status of children aged 6‐59 months (n=355) living in such communities. Food security was assessed using a 6‐item validated food security survey module. We found significantly higher rates of stunting in severely food insecure households (SFI) (23.4%) than food secure (4.9%) and moderately food insecure households (6.1%). After adjusting for age, gender and socio‐economic status, severe food insecurity remained an independent predictor of stunting (OR=3.74, p=0.002). Prevalence of overweight was also higher in the SFI, but this difference did not reach statistical significance. In the context of the advancing nutrition transition in Lebanon and other Middle Eastern countries, the results of this study indicate the need to explore and address the underlying causes of household food insecurity. This could contribute to reducing risk of childhood stunting, and its potential long term consequences on child health and development.

Grant Funding Source: Supported by the American University of Beirut, the Heinrich Boll Foundation and UNHCR.
Longer breastfeeding reduces the positive relationships among gestational weight gain, birthweight, and childhood growth (1017.11)
Authors: Zhu Yeyi , Hernandez Ladia , Dong Yongquan , Himes John , Hirschfeld Steven , Forman Michele
Published: June 2021

Despite the well‐studied relationship between gestational weight gain (GWG) and childhood obesity, little is known about whether breastfeeding protects against obesity through GWG. We investigated the mediation effect of breastfeeding duration on the relationships among GWG, birthweight, and childhood growth in 1464 mothers and their children aged 0‐6 y in the National Children’s Study in Anthropometry. Child growth parameters included weight‐for‐age Z‐score (WAZ), height‐for‐age Z‐score (HAZ), weight‐for‐height Z‐score (WHZ), and body mass index Z‐score (BMIZ) based on CDC growth references. Structural equation modeling assessed both the direct and indirect effects of GWG on growth. After adjusting for maternal sociodemographic and perinatal covariates, GWG had significant direct effects on HAZ, WHZ, and BMIZ [path coefficients (β) = 0.055, 0.061, and 0.084, respectively]. In addition, birthweight mediated the positive relationships between GWG and all growth parameters whereas breastfeeding duration had negative effects on WAZ (β = ‐0.067, P < 0.01) and BMIZ (β = ‐0.09, P < 0.05). Thus, longer breastfeeding reduces the positive relationships among GWG, birthweight, and WAZ/BMIZ. Given that greater effects of breastfeeding duration were observed with increasing GWG, these findings underscore the importance of promoting breastfeeding following pregnancy to mitigate childhood obesity.

Grant Funding Source: Supported by HHSN275200800020C
Comparative analysis of gut microbiota of breast‐ and formula‐fed Korean infants by using pyrosequencing (1017.4)
Authors: Lim Ji Ye , Lee Sang A , Rhie Mi Na , Kim Bong‐Soo , Cho Su Jin , Kim Ok Bin , Kim Yuri
Published: June 2021

Infant feeding is one of the significant determinants of the intestinal microbiota in early life. Although numerous researches have performed to investigate the bacteria community in the infant gut, culture‐based methods had showed limited bacteria population. Recently, pyrosequencing, a new high throughtput DNA sequencing technique, has been introduced and applied to explore microbiome. The purpose of this study was to compare the compositions of the fecal microbiota of Korean infants fed breast milk to those of infants fed milk formula. We examined microbial communities in fecal samples collected from 20 Korean infants at 4 weeks of age using pyrosequencing. Diversity analysis of total microbiota sequences revealed that a more diverse collection in milk formula‐fed infants (MFI), whereas, the probiotics, including S. salivarius, L. gasseri, and B. longum were abundant in the microbiota of breast milk‐fed infants (BFI). Moreover, Bifidobacteriaceae were significantly abundant in BFI compared with those of FFI. Our study is the first attempt to analyze gut microbiota in Korean infants by using pyrosequencing, which may provide valuable information to future study of the role of intestinal microbiota affecting in adulthood.

Grant Funding Source: Ewha Womans University Research Grant of 2013.
Maternal attitudes toward human milk sharing (1017.7)
Authors: O'Sullivan Elizabeth , Geraghty Sheela , Rasmussen Kathleen
Published: June 2021

Sharing of expressed human milk (HM) is becoming increasingly common in the US. Little is known about mothers’ opinions of this practice. Our goal was to gain a deeper understanding of maternal attitudes toward HM sharing. We conducted retrospective interviews (n = 30) with a diverse sample of mothers of children aged ~1‐3 years. Women were asked how they felt about HM sharing generally as well as their personal experience of providing or obtaining HM. Interviews were transcribed and themes were identified using an inductive approach. Several women had provided HM for another infant and some had obtained HM from another woman. Providers often had excess HM that they did not think that their infant would ever consume. Obtainers either did not produce enough HM or they had another medical condition affecting their ability to feed their own HM. Many interviewees were open to the idea of providing HM for another infant; however, fewer said they would be willing to obtain HM from another woman. Many women, whether they obtained HM or not, expressed concern about the dietary habits or disease status of HM providers; only a few mentioned hygienic HM expression and handling. It was generally assumed that, if the HM was being consumed by the provider’s own child, then it must be safe. Given the potential for microbial contamination of expressed HM, the extent of HM sharing and its safety must be determined.

Grant Funding Source: EOS was supported by an International Fulbright Science & Technology Award
Yogurt consumption, socio‐demographic factors and lifestyle of the Italian population: results from the INRAN‐SCAI 2005‐06 food consumption survey (1018.2)
Authors: Turrini Aida , D'Addezio Laura , Mistura Lorenza , Sette Stefania
Published: June 2021

Yogurt consumption and associated parameters were analysed on a subsample of 2831 subjects 蠅18 years from the INRAN‐SCAI 2005‐06 survey. Association of yogurt consumption with socio‐demographics, smoking and alcohol habits, physical activity, dieting, regular breakfast intake, eating out habits, and interest for nutrition information was evaluated through logistic regression analysis.

Mean intake of yogurt, based on 3 consecutive days, was 21 g/day, with 24% of consumers who had a mean intake of 88 g/day. Females were twice as likely to consume yogurt. Likelihood of consumption was higher for 18‐64 vs. 蠅65 year olds, Northern & Central vs. Southern regions, and for higher (蠅2 h/week) vs. lower (<2 h/week) physical activity. Smokers were 30% less likely to consume yogurt than non‐smokers. Regular breakfast consumers were 2.7 times more likely to eat yogurt than those who skipped it, knowing that breakfast was the moment of consumption for which highest intake was registered (8.3 g/day vs. 4.8 g/day for snacking). Those who reported to often eat out showed a reduced tendency to consume yogurt.

Finally, yogurt consumers also showed a positive trend towards a better diet quality (PanDiet score) and increasing interest in nutritional information.

In conclusion, Italian population has a low consumer rate of yogurt but data suggest that consumption of this product is associated with several healthy lifestyle parameters.

Grant Funding Source: Danone Nutricia Research, Centre Daniel Carasso
Children’s liking of child‐friendly shaped fruits and vegetables: does shape influence liking? (1019.22)
Authors: Baker Selena , McCabe Sean , Swithers Susan , Payne Collin , Kranz Sibylle
Published: June 2021

Fruit and vegetable intake is low in most children. Children’s liking of foods predicts intake; therefore, increasing liking of fruits and vegetables (FV) may be an initial step in increasing consumption. The aim of this study was to explore whether offering FV in child‐friendly (CF) shapes increases children’s liking of these foods. A convenience sample was recruited for a survey and optional taste‐test. FV were randomly offered in regular shapes (chunks/slices) or “fun” CF shapes (butterfly, chick, flower, teddy bear). Participants (n=365) were 34% more likely to select CF vs. regular‐shaped FV. Liking of CF‐shaped foods was high (median=5 on 5‐pt. scale) and they were seen as more fun than regular‐shaped foods (p<0.001). Sensory ratings were not significantly different between shapes. Participants were more likely to choose fruits than vegetables. Our results suggest that the shape of FV influences how fun children perceive them to be and how likely they are to select them for a taste‐test, but does not impact liking. Offering FV in CF shapes may increase children’s motivation to select these foods. Future research should examine a wider variety of foods in order to elucidate these effects.
Development of child‐friendly fish dishes to increase young children’s acceptance and consumption of fish (1019.24)
Authors: Huss Lyndsey , McCabe Sean , Dobbs‐Oates Jennifer , Burgess John , Behnke Carl , Santerre Charles , Kranz Sibylle
Published: June 2021

Although fish and algae are the only dietary sources of docosahexaenoic acid (DHA), a nutrient critical for brain development, fish is vastly under‐consumed in children. Plant‐based omega‐3 fatty acids can be converted to DHA but the conversion is inefficient. The Dietary Guidelines for Americans recommend consumption of seafood twice a week. We examined if substituting salmon for chicken in childcare center lunch meals could increase preschoolers’ fish and DHA consumption. Participants ages 2 ‐ 5 (n=45) were served eight study lunches (four chicken or salmon dishes) once a week over 8 weeks. Analysis of plate‐waste showed no significant differences in energy intake when the foods looked similar, but when the fish dishes looked different, energy intake on “fish days” was lower than on the corresponding “chicken day.” However, DHA intake increased significantly on all “fish days”. These results indicate that fish and DHA intake can be effectively increased if fatty fish, such as salmon, is used as a substitute for already accepted protein sources in the childcare center menu.
Nutrient intakes among infants and children from birth to 3 years (1023.1)
Authors: Storey Maureen , Anderson Patricia
Published: June 2021

The Dietary Guidelines has focused on adults and children 2 years of age and older. However, the 2020 Dietary Guidelines will address Americans of all ages, starting from birth. We examined nutrient intakes of children from birth to 6 months, from 6 months to 12 months, and from 1‐3 years using the most recent publicly available dietary data from the National Health and Nutrition Examination Survey 2009‐2010. We chose these age groups to align with the age groups used for the National Academies of Sciences Dietary Reference Intakes. Appropriate survey weights were used to calculate average daily intakes of protein, total fat, dietary fiber, calcium, phosphorus, magnesium, and potassium. On average, most infants from birth to 6 months and from 6 to 12 months met the existing National Academies’ nutrient recommendations. However, children 1 to 3 years of age did not consume adequate amounts of potassium or dietary fiber, on average. In general, children 1‐3 y consumed 10.3 g of dietary fiber while the Adequate Intake (AI) level is 19 g. Similarly, children 1‐3 y consumed 2024.8 mg of potassium, on average, which is nearly 1000 mg short of the established AI for potassium. The inadequacies were consistent for both males and females and across different race/ethnicities. These results suggest that increasing potassium and dietary fiber intake among young children should be a priority for the 2020 Dietary Guidelines.
Recent decreases in beverage calories among U.S. preschoolers slowed by economic recession (1023.3)
Authors: Ford Christopher , Ng Shu Wen , Popkin Barry
Published: June 2021

NHANES data show decreases in caloric intake in preschool children from 2003 to 2010. The effect of the Great Recession on beverage intake trends is unclear, as the cross‐sectional NHANES analyses cannot control for economic factors. Thus, we compare trends in preschooler beverage intakes in NHANES to trends in beverage purchases, adjusted for socioeconomic factors, among households with preschoolers in Nielsen Homescan.

We included 3,358 preschoolers (age 2‐5y) from NHANES 2003/04 to 2009/10, and 14,700 households with preschoolers from Nielsen Homescan 2003 to 2012. For total beverages, juice/juice drinks, sodas, plain milks, and sweetened milks, we computed mean intakes per capita/d using NHANES data, and adjusted per capita/d purchases using Homescan data and random effects models controlling for household composition, demographics, market‐quarter unemployment rates and food/ beverage prices.

Between 2003 to 2010, unadjusted per capita intake and purchases for total beverages fell by an annual average of 2.7% and 3.4% respectively. After adjusting for economic factors, total beverage purchases fell by an annual average of 6.6% between 2003 and 2010. For juice/juice drinks, plain milks and sweetened milks, average annual decreases in beverage purchases were more slightly more pronounced than those for beverage intakes. After adjusting household beverage purchases for socioeconomic factors and prices, average annual decreases in household purchases for all beverage groups (including total beverages) were even more pronounced than unadjusted household beverage purchases. Adjusted annualized decreases in household beverage purchases from 2009/10 to 2011/12 were similar to those of prior years.

Decreases in beverage purchases, adjusted for socioeconomic factors, were more pronounced than unadjusted decreases in beverage intakes and purchases, suggesting that the Great Recession attenuated a secular decline in preschooler beverage trends that persisted through 2012.

Grant Funding Source: Supported by RWJF (Grant 70017), NIH (R01DK098072) and CPC (5 R24 HD050924)
Micronutrient intakes among U.S. children: understanding variability in different age and gender groupings in NHANES 2007‐2010 (1024.13)
Authors: Eldridge Alison , Rytz Andreas , Van der Horst Klazine , Fulgoni Victor
Published: June 2021

The Institute of Medicine (IOM) presents Dietary Reference Intake recommendations according to specified age, gender and life‐stage groups, including 1‐3 years and 4‐8 years for children, and 9‐13 years and 14‐18 years separately for males and females. Researchers, however, often present dietary intake data using different groupings. This project used data from the U.S. National Health and Nutrition Examination Survey (NHANES) 2007‐2010 to evaluate the variability in reported intakes of micronutrients in children using various age and gender cut‐offs. The IOM groupings were used, but since dietary intake data from NHANES are not available for 1‐year‐olds, a grouping for 2‐3 year olds was used instead. In addition, intake data for 4‐6 year olds, 7‐9 year olds, 10‐12 year olds, 13‐18 year olds and 4‐12 year olds were also evaluated for both genders combined and for males and females separately. The National Cancer Institute method was used to adjust the NHANES dietary intake data to provide more reliable estimates of habitual consumption (NCI, 2012). Variability of micronutrient intakes and percentage of the population below recommended intakes within each age and gender category were evaluated using z‐scores. Finding the most relevant age groups to present dietary intake data is important because the type and amount of foods eaten change throughout growth and development. Age categories should reflect these changes to best understand dietary patterns.

Grant Funding Source: Nestle Research Center
Comparison of childhood cancer survivors’ dietary intake with U.S. dietary guidelines (1024.2)
Authors: Zhang Fang Fang , Saltzman Edward , Must Aviva , Liu Shanshan , Kelly Michael , Parsons Susan , Roberts Susan
Published: June 2021

Despite improved survival, childhood cancer survivors are at a much higher risk of developing chronic health conditions than their siblings. Poor diet quality increases chronic health risk but the diet quality of childhood cancer survivors has not been adequately studied. We assessed the Healthy Eating Index (HEI) in 22 childhood cancer survivors and compared survivors’ dietary intake to the 2010 Dietary Guidelines for Americans (DGA). Dietary data were collected using repeated 24‐hour diet recalls over a one‐year study period. The mean HEI 2010 in childhood cancer survivors was 52.7. Long‐term survivors who were off‐treatment 蠅10 years had a significantly lower HEI than recent survivors who were off‐treatment< 5 years (β = ‐11.5, 95% CI: ‐22.1, ‐0.9, p=0.047). For individual nutrients, none of the survivors met the guidelines for dietary fiber and potassium intake. Only 29%, 24%, 19% and 4% met the guidelines for saturated fat, calcium, sodium and vitamin D intake. The average intake in relative to the guidelines was 63% for fiber, 85% for calcium, 32% for vitamin D, and 50% for potassium, but was 115% for saturated fat and 143% for sodium. Childhood cancer survivors, in particular long‐term survivors, have a poor diet quality as determined using the HEI 2010. Targeted dietary interventions are needed to address the poor diet quality in childhood cancer survivors early during survivorship.

Grant Funding Source: P30DK46200, UL1RR025752, UL1TR000073
Maternal high fat feeding alters bone lipid content at weaning without long‐lasting effects on bone lipid content and bone strength in male offspring at young adulthood (1033.7)
Authors: Miotto Paula , Castelli Laura , LeBlanc Paul , Peters Sandra , Roy Brian , Ward Wendy
Published: June 2021

High fat (HF) feeding after weaning results in elevated bone fatty acid content and reduced bone strength in growing rats and mice. However, the effect of a maternal HF diet to offspring in utero and during suckling is less characterized. Our study objective was to determine if maternal HF feeding altered male offspring bone lipid composition at weaning and 3 months of age, and if this was accompanied by lower bone strength. Female Wistar rats were fed either control (CON, 7% soybean oil by weight (TD.94045); n = 9) or HF (20% lard by weight (TD.02016); n = 9) diet for 10 weeks. Rats were bred and consumed their respective diets throughout pregnancy and lactation. All offspring were weaned to CON diet. At weaning, but not 3 months of age, percent mole fraction of polyunsaturated fatty acids were lower and monounsaturated fatty acids were higher in femur of offspring from dams fed HF diet than offspring of dams fed CON diet (P <0.05). Peak load, the maximum force a bone withstands before fracture, was not different between groups at the femur (midpoint and neck) and 3rd lumbar vertebra at 3 months of age. In conclusion, in utero and suckling exposure to a HF diet does not have long‐lasting effects on bone health in this rat model. Moreover, at 3 months of age, lipid composition of femurs no longer reflects maternal HF feeding and may explain why bone strength is similar between groups.
Postprandial inflammatory responses and plasma‐free fatty acids: effects of blueberry intake (1034.6)
Authors: Ono‐Moore Kikumi , Snodgrass Ryan , Huang Shurong , Singh Shamsher , Burnett Dustin , Bonnel Ellen , Woodhouse Leslie , Freytag Tammy , Zunino Susan , Rutledge John , Hwang Daniel
Published: June 2021

We studied whether a high‐fat (HF) meal‐induced postprandial (PP) lipemia affects inflammatory responses in blood monocytes, and whether blueberry intake suppresses the PP inflammatory responses in healthy human subjects. A HF breakfast increased PP plasma triglycerides and insulin, but non‐esterified free fatty acid (NEFA) levels were decreased compared to fasting levels. TNF‐α , IL‐6 and IL‐8 in PP plasma were decreased compared to fasting levels. Cytokine and NEFA levels in PP plasma were not affected by blueberry intake. To determine whether endogenous NEFAs derived from the HF diet stimulate monocyte activation, whole blood or peripheral blood mononuclear cells (PBMC) were treated with lipoprotein lipase (LPL). LPL treatment of whole blood increased NEFA levels and secretion of IL‐1β, IL‐6, and TNF‐α. IL‐1β in LPL‐treated PP whole blood was elevated compared to fasting control. Blueberry intake suppressed IL‐1β and IL‐6 production without affecting levels of NEFAs released in LPL‐treated PP blood when fasting controls were used as a covariate. LPL treatment also increased NEFA and cytokine levels in PBMC. Together, these results demonstrate that NEFA levels in plasma or whole blood are an important determinant affecting pro‐inflammatory cytokine production. Therefore, modulation of levels of plasma NEFA by dietary means could be an effective strategy to suppress HF diet‐induced inflammatory responses.

Grant Funding Source: Supported by USDA‐ARS‐WHNRC Funds and US Highbush Blueberry Council grant
Theobromine suppresses adipocyte differentiation through the MAPK pathway in 3T3‐L1 preadipocytes (1037.11)
Authors: Jang Yeonjeong , Kim Jiyun , Lee Heeweon , Pyo Suhkneung
Published: June 2021

Theobromine (TB) is an alkaloid belonging to the methylxanthines. It is mainly found in cocoa beans. In this study, we show that the inhibitory effect of TB on adipocyte differentiation in 3T3‐L1 cells by reducing adipogenic gene expression. To investigate the effects of TB on adipocyte differentiation, 3T3‐L1 cells were induced to differentiate using 3‐isobutyl‐1‐methylzanthine, dexamethasone, and insulin (MDI) for 8 days in the presence of 50, 100, and 150 μg/ml TB. TB attenuated the accumulation of lipid droplets stained with Oil Red O in dose‐dependent manner. We also found the effect of TB on the inhibition of adipocyte differentiation occurred during early stage of adipogenesis. To elucidate the anti‐adipogenic mechanism of TB, adipogenic transcription factors, the phosphorylation levels of MAPK, and adipogenic gene expression were analyzed by Western blotting. Treatment of cells with TB resulted in a decrease in the protein level of (Peroxisome proliferator‐activated receptor gamma) PPARγ, (CCAAT/enhancer binding protein beta) C/EBPβ, and phosphorylation of ERK and JNK. These results demonstrate that TB inhibits adipogenic differentiation via suppression of the ERK and JNK signaling pathway during the early stages of adipogenesis.
Microflora changes with an acute high fat diet (1037.13)
Authors: Gonzalez Christopher , Haar Lauren , Engevik Melinda , Jones W.
Published: June 2021

Activation of NF‐κB signaling and ensuing protective gene transcription in the heart is the basis for cardioprotection in the murine heart following an acute (24h) high fat diet (HFD). This protection is strongest after 2wks of high fat feeding and declines thereafter. The exact connection between acute high fat feeding and NF‐κB activation is not yet fully understood. Long‐term fat absorption in the intestine is associated with increased vascular permeability, decreased numbers of gram‐positive bacteria, and cytokine release into lymphatic circulation. We considered the novel hypothesis that an acute HFD affects the intestinal microflora. Bacterial DNA from stool samples, luminal flushes, and mucosal scrapings were isolated from 24h high fat fed mice and changes in the phyla Firmicutes, Actinobacteria, Bacteroidetes, and Y‐Proteobacteria measured by quantitative polymerase chain reaction. Our data indicates a significant increase in Actinobacteria in stool post high fat feeding. No statistically significant differences were found among bacteria in the flushes or mucosa at the 24hr time point. Conceivably, activation of pattern recognition receptors, or increased release of immune modifiers such as cytokines, known activators of NF‐κB systemically, could result from the observed changes in GI microflora post acute HFD. Our pilot data suggests focusing on cytokine regulation in response to short term high fat feeding. Further experimentation to identify the changing strains of Actinobacteria and quantify levels/activation of TRL4 and IL‐6 post 24h HFD, as well as experiments to directly link the changes in flora to cardioprotection are underway.
The effect of solid, semi‐solid and fluid snacks on food intake and satiety in children (1040.6)
Authors: Li Athena , Gheller Brandon , McCormick Mary , Bellissimo Nick , Anini Younes , Nunes Fernando , Hamilton Jill , Mollard Rebecca , Anderson G. Harvey , Luhovyy Bohdan
Published: June 2021

The increase in the frequency of snacking and the types of snacks consumed among children may contribute to metabolic disorders including obesity and diabetes. The objective of this study was to explore the short‐term effects of dairy and non‐dairy snacks on subjective satiety and food intake in children. Methods: In a repeated‐measures design, 23 normal weight (5th‐85th BMI percentile) children (16 girls and 7 boys; aged 9‐14 y) were randomly assigned to one of five isocaloric (180 kcal) treatments including solid foods: deep fried potato chips (32g), mini sandwich type cookies (39g), and mozzarella cheese (63g), semi‐solid: Greek yogurt (200g), and fluid: milk (2% milk fat, 346g). Following a 12hr overnight fast, participants consumed a standardized breakfast two hours prior to each study session. Subjective appetite was measured with visual analogue scale questionnaires before and at 15, 30, 45, 90, 120 and 145 min after the treatment. Food intake was measured with an ad libitum pizza meal provided at 120 min. Results: Food intake was 82 kcal lower after the cheese treatment compared to the milk treatment (P<0.05) while the other treatments led to the intermediate results. There was an effect of time (P<0.0001), but no treatment or time‐by‐treatment effect on subjective average appetite. Conclusion: A solid dairy snack (cheese) is effective in reducing food intake in children.

Grant Funding Source: Dairy Farmers of Canada
Vitamin D Intake among adults of varying age, race, and gender (1041.1)
Authors: Bailey Alison , Gonzales Dana , Crook Tina , Hakkak Reza
Published: June 2021

Vitamin D deficiency is a significant problem in the United States, especially among older adults. The objectives of this studywere to determine if age, gender, and race influence vitamin D intake and to determine if varying degrees of vitamin D inadequacy exist in adults. Secondary data analyses were performed on 1,065 adults (>41 years) in the 2009‐2010 NHANES data set. We found that Mexican Hispanics had significantly higher vitamin D intake levels compared to Non‐Hispanic Whites (p=.003, 2‐tailed). Analysis also showed that there was a significant difference in the vitamin D intake levels among males (0.6 ‐ 38.6 mcg/d) and females (5.6 ‐ 43.2 mcg/d), p<.001. There was no relationship between vitamin D intake and age. The findings suggest that the mean intake of vitamin D for age, race, and gender where all above the RDA (15 mcg/d), but the min ‐ max ranges for all participants in the study suggest some individuals were well above the RDA for vitamin D while others were well below that threshold. Much of the current research on vitamin D is showing a correlation between deficiency and all‐cause mortality. Further research is needed to confirm this correlation.
The bioavailability iron, zinc and copper in the rat due to their interactions (1042.8)
Authors: Gordon Dennis , Ellersick Mark
Published: June 2021

There are many factors believed to influence nutrient bioavailability. However, possibly the most significant factors affecting the bioavailability of Fe, Zn and Cu, because of their similar chemical nature, are their levels in the body and dietary concentrations. The ultimate objective and results of this interaction study in rats and related studies in pigs and humans were used to formulate foods with optimum mineral concentrations to achieve optimum bioavailability. First determining the threshold levels of Fe, Zn and Cu needed to sustain adequate status in weanling rats, 36 groups of 5 animals per groups, were randomly arranged in a 3 (Fe) X 4(Zn) X 3(Cu) factorial. While all nutrients were constant in the AIN‐76 diet, dietary levels in ppm were: Fe, 18, 90 and 270; Zn, 8.5, 10.5, 42.5 and; Cu, 2, 10 and 40. Mineral status among all groups after 28 days ad libitum feeing was assessed by Hb levels, and liver and femur Fe, Zn and Cu concentrations. Significant interactions were observed for replication, Fe, Zn, Cu, Fe*Zn, Fe*Cu, Zn*Cu and Fe*Zn*Cu and with few exceptions all at P蠄0.01. The most significant and important interaction among Fe*Zn*Cu were/are illustrated using response surface methodology. Taking into consideration the most appropriate level of one element to be added to a food, estimates for the other two elements at that level could be determined and the process can be completed/repeated for all three elements.

Grant Funding Source: Missouri Agricultural Experiment Station