bluuebunny/crossref_metadata_2025 huggingface dataset:
Visualizing Biochemistry: The Implementation of Biomolecular Visualization and Structure‐Function Conceptual Game‐based Card Sorting Activities
Authors: Bates Alison , Terrell Cassidy R.
Published: June 2021

Biochemistry, at the undergraduate level, is in a distinctive position to unveil misconceptions as the topics covered are abstract and require recollection of content from prior STEM courses. These components, when compounded with myriad of representations, likely exceed the cognitive load of the students and therefore propagate misconceptions. Current research suggests that undergraduate students learn visual literacy skills indiscriminately throughout his/her academic career. Therefore, we propose that direct instruction of biomolecular visualization will reduce the cognitive load and increase conceptual understanding.

This study aims to answer the questions: Do targeted interventions improve accuracy and precision of card sorting across the semester? and Do targeted interventions increase conceptual understanding of structure and function across the semester? Preliminary findings from pre‐ and post‐ “free sort” card sorting activities in control semesters provided evidence for the design of a series of interventions. These interventions aim to increase the visual literacy of undergraduate biochemistry students while simultaneously increasing structure‐function content understanding using game‐based card sorting activities. Game‐based learning is used to increase motivation, comprehension, and sense of achievement in students. Three interventions were designed and implemented into the biochemistry curriculum for fall 2017. Student responses from these activities and the matching card section on exams were analyzed in addition to the pre‐ and post‐”free sort” card sorting activities. These data are compared to control groups from two previous semesters. Furthermore, the data are analyzed using qualitative inferential, quantitative inferential, and descriptive statistical analysis. Refinement of the interventions will continue into future semesters of biochemistry with the intention of setting a standard of targeted instruction of visual literacy skills.

Support or Funding Information

This project is partially supported by the National Science Foundation under award number DUE1323414 to Milwaukee School of Engineering.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using developmental rate variation to establish a fine‐grained sequence of ossification center differentiation
Authors: Flaherty Kevin V. , Halmi Caroline , Richtsmeier Joan
Published: June 2021

Mouse embryos from inbred strains exhibit a large range of phenotypic diversity, despite tight controls placed on genetic variation and environmental conditions. Individuals vary in terms of body size and the progress of developmental morphology, even within litters. This indicates that individual embryos develop at different rates. Staging atlases such as those developed by Kaufman and Theiler provide details of the morphological features and developmental processes evident on successive days of development, but do not offer a systematic method for determining if two embryos represent similar developmental stages within more fine‐grained time periods. To ensure that studies of developmental processes like tissue interactions and gene expression events that occur within restricted periods of a few hours are not confounded by inter‐individual variation in rates of development, precise temporal control is needed to sequentially order and contextualize data collected from individual embryos. We collected and compared temporal sequences of ossification center appearance and development using two measures of individual development: size, measured as crown‐rump length, and shape, using the eMOSS staging system (http://www.limbstaging.crg.es/) that generates estimates of individual ages based on the 2D outline of the hindlimb.

Using crown‐rump length and the eMOSS system, we demonstrate significant developmental variation within litters, and significant disparity between an embryo's age based on the age at collection and age generated from eMOSS. Using eMOSS, we are able to elucidate the sequence of bone development in unprecedented detail. We recorded the appearance of ossification centers by scoring the presence of the critical osteoblast differentiation gene Osterix in OSX‐GFP mice sacrificed at 6 hour increments from E12.0 to E15.0 and scored the appearance of calcified bone by staining C57Bl/6 embryos between E14.0 and E16.0 with calcein. We improved the understanding of the temporal sequence and precise timing of these developmental events by generating an ossification center development sequence that details the appearance of bones according to the overall body size, and more importantly, the developmental hour at which they appeared.

Support or Funding Information

This work is supported in part by the NIH: Craniosynostosis Network P01HD078233 & R01DE018500, R01DE022988; NSF BCS‐1650824; and the Burroughs Wellcome Fund.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early‐Life Sickness May Predispose Individuals to Behavioral Changes Following Gut Dysbiosis in Adulthood
Authors: Sylvia Kristyn E. , Deyoe Jessica E. , Demas Gregory E.
Published: June 2021

Although it is well‐established that the immune system plays an important role in the development of physiology and behavior, the gut microbiome has recently become of interest in the study of developmental origins of behavior. Some studies suggest that the effects of early‐life immune activation may not occur until a secondary stressor is introduced, though the precise nature and timing of the stressor may be critical in the response. In order to test the hypothesis that early‐life immune stress makes an individual more vulnerable to future insults, we mimicked an early‐life infection by injecting pups at postnatal day 3 and 5 with lipopolysaccharide (LPS; cell wall component of gram‐negative bacteria) or saline, and subsequently exposed the same animals to antibiotic treatment (known to influence microbial community composition ad behavior) or water in adulthood. We tracked physiology across development, and paired males and females with a novel individual of the same sex in adulthood to score behavior before antibiotic treatment, immediately following treatment, and after recovery from antibiotics. LPS‐treated females exhibited impaired reproductive physiology and function in adulthood (e.g., smaller ovaries and abnormal estrous cycles), and female and male gut microbial communities were strongly affected by antibiotic treatment in adulthood, but only slightly affected by postnatal LPS alone. Interestingly, LPS‐treated males exhibited more robust changes in their behavioral response following adult antibiotic treatment, including decreased investigation and increased grooming. These data suggest that males may be more vulnerable than females to behavioral abnormalities after being predisposed to an immune challenge early in life. Collectively, these results provide evidence that some of the sex‐specific behavioral consequences of a stressor may not transpire until an individual is faced with a secondary stressor, and the context in which an individual is exposed can greatly influence the response.

Support or Funding Information

National Institute of Child Health and Human Development (T32HD49336); National Science Foundation, Division of Integrative Organismal Systems (1656414); and Indiana University

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying C‐Mannosylated Proteins in RAW264.7 Cells
Authors: Kegley Nicholas Ryan , Boober Karli , Ito Atsuko , Takeuchi Megumi , Haltiwanger Robert S , Rakus John
Published: June 2021

C‐mannosylation is a specific type of glycosylation in which α‐mannose is covalently bound to the C2 atom of a tryptophan via a carbon‐carbon bond. Presently, it is known to occur on proteins that contain the W‐x‐x‐W‐x‐x‐W/C motif found within the Thrombospondin Type‐1 Repeat (TSR), with ‘x’ denoting any other amino acid. C‐mannosylation has been implicated in a few cellular functions, including protein secretion and type I cytokine receptor function. Previous research has shown that exogenous C‐mannosylated peptides derived from TSR containing proteins upregulate the production of the pro‐inflammatory cytokine Tumor Necrosis Factor alpha (TNF‐α) in RAW264.7 macrophage cells. This upregulation is substantially increased in the presence of Hsc70. When these peptides are internalized by RAW264.7 cells, a binding interaction occurs between the two proteins, which also correlates with increased TNF‐α levels. With the knowledge that exogenous C‐mannosylated peptides and Hsc70 can influence cytokine production, this study has focused on identifying endogenous C‐mannosylated proteins in RAW264.7 cells that interact with Hsc70. The potential of a naturally occurring interaction between these two proteins could elucidate the importance of C‐mannosylation in the innate immune response. Through a Hsc70 co‐immunoprecipitation, a potentially C‐mannosylated protein has been identified in Junctophilin‐1 (JPH1). JPH1 forms a junctional membrane complex (JMC) between the endoplasmic reticulum and plasma membrane of excitable cells, facilitating communication between calcium ion release channels. When comparing the nano‐LC MS/MS results for JPH1 against a mouse database, 69 separate PSMs were shown where JPH1 carries a hexose. MS/MS data suggests that the hexose appears on a single tryptophan in a peptide bearing the sequence: E‐G‐E‐W‐A‐N‐N‐K. This indicates that JPH1 may be an example of non‐canonical C‐mannosylation. To further explore the status of JPH1 as a C‐mannosylated protein, the protein was overexpressed in HEK293T cells. This experiment showed that JPH1 was not C‐mannosylated in HEK293T cells. Noting the lack of modification, RTPCR was performed with both RAW264.7 and HEK293T RNA. These experiments showed that all four DPY19L homologs were expressed in HEK293T cells, while only DPY19L1 and L4 were expressed in RAW cells. Further work is being conducted to overexpress JPH1 in RAW264.7 cells to determine why, or if, JPH1 is C‐mannosylated in this particular cell line.

Support or Funding Information

This work was supported by NSF grant OIA‐1738707.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An ex vivo splenic explant model system for the identification of small molecule therapeutics for visceral leishmaniasis
Authors: Osorio Yaneth E. , Travi Bruno L. , Melby Peter C.
Published: June 2021

New therapies for leishmaniasis are needed. We established an ex vivo splenic explant culture system from hamsters infected with luciferase‐transfected Leishmania donovani to screen chemical compounds for leishmanicidal activity. This model has advantages over in vitro systems in that it: 1) includes the cell populations involved in the host‐parasite interaction, 2) is initiated when the permissive immune mechanisms that lead to progressive disease are evident; 3) supports active parasite replication that can be easily measured by detection of parasite‐expressed luciferase; and 4) can be used to identify compounds that have both direct and immune‐mediated activity. The assay showed excellent discrimination between positive and negative controls (Z’ Factor >0.8). A screen of 1,040 small molecules identified 57 hits (Z score > 1.96; p<0.05). One‐third of the compounds were identified by database search as having toxicity; one‐fourth were administered previously to humans. The results were consistent across a 2.5–10 μM concentration range. Further testing of these lead compounds may identify new drugs for the treatment of leishmaniasis. Supported by the Department of Defense.
Comparing the susceptibility of genetically modified mice to intoxication by organophosphorus nerve agents
Authors: Cadieux C. Linn , Guzman Juanita J. , Corun Charlene M. , Rausa Rebecca C. , Lenz David E. , Cerasoli Douglas M.
Published: June 2021

To examine the contributions of butyrylcholinesterase (BuChE) and paraoxonase 1 (PON1) to the gestalt resistance of mice to organophosphorus (OP) nerve agents, we determined the LD50s of GB, GD, and VX in mice deficient in BuChE, PON1, or both; likewise, mice expressing either human PON1 or the mutant G117H human BuChE were tested. The results indicate that the absence of BuChE, PON1, or both does not enhance susceptibility to GB or GD, and has only a modest effect on susceptibility to VX (statistically significant only for female mice). The transgenic expression of human PON1 or G117H BuChE did not enhance resistance to OP intoxication. LD50s varied by as much as 50% between mice of different backgrounds, independent of the presence of knockout or transgene alleles. Female mice of all backgrounds were more resistant to OP intoxication than corresponding males. The results suggest that in mice, neither BuChE nor PON1 plays a substantial role in resistance to OP intoxication. Further, for these enzymes to provide protection if administered as exogenous bioscavengers, they must be present at concentrations higher than normally found in mice.

This work was supported by funding from DTRA.
The relation between serum xanthophylls, fatty acids, macular pigment and cognitive function in the Health ABC Study
Authors: Renzi Lisa Marie , Iannaccone Alessandro , Johnson Elizabeth , Kritchevsky Stephen
Published: June 2021

Xanthophylls lutein and zeaxanthin (L, Z) and ù‐3 fatty acids (FAs), are optimally positioned in retina and cortex to protect from oxidative damage. Research suggests that this placement in retinal tissue is related to reduced risk for acquired retinal diseases and improved visual function (e.g., reduced glare disability, scotopic noise). Whether L, Z and FAs in the cortex are capable of similarly influencing cognitive function (CF) is unknown. The purpose of this investigation was to determine the relation between L, Z and FAs and CF in a healthy cohort of older adults. 118 subjects in the Memphis, TN area, aged 76–85 yrs, with approx. equal numbers of males/females. Retinal L, Z levels (i.e., macular pigment density, MPOD) were significantly related to performance on a variety of indices designed to assess processing speed, accuracy and completion ability (p≤0.05). These relations remained significant after adjusting for age, sex and ethnicity. Serum xanthophyll and fatty acid levels were not related to CF. MPOD, unlike serum xanthophylls, is a reflection of L and Z embedded into CNS tissue. The relation between MPOD and CF suggests that L and Z embedded in cortical tissue are capable of influencing cortical function. Additional research will be necessary to confirm these relations.

Support:USDA#581950‐7‐07; DSM; AHAFMDR; NIHK23EY000409; N01AG62101; N01AG62103; N01AG62106; IRRF; RPB
Improved nutrient intake and diet quality associated with lean beef consumption in the US: National Health and Nutrition Examination Survey (NHANES) 1999–2004
Authors: Zanovec Michael , O'Neil Carol E , Keast Debra R , Fulgoni Victor L , Nicklas Theresa A
Published: June 2021

The dietary guidelines recommend consuming meats in its lowest fat form. NHANES 1999–2004 24‐hr dietary recall data were used to compare nutrient intake and diet quality (HEI‐2005) between highest lean/lowest fat (HLLF) beef consumers and lowest lean/highest fat (LLHF) beef consumers aged 4+ y (n = 11,507). Beef intakes were determined by summing amounts of beef contained in survey foods. The fat content of beef consumed was linked to the survey‐specific Nutrient Database for Standard Reference. Beef consumers were classified into tertile groups based on the fat content of beef consumed. HLLF beef consumers had higher protein intake; lower energy; total, saturated, and monounsaturated fat; and carbohydrate than LLHF beef consumers. HLLF beef consumers also had significantly higher intakes of vitamins A, C, B3, B6, B12; phosphorus; magnesium; iron; zinc; and potassium; and lower intake of calcium than LLHF beef consumers. Compared to the LLHF beef group, HLLF beef consumers also had higher cup equivalent intakes of total vegetables, dark green/orange vegetables, and legumes, higher meat oz equiv, lower grain oz equiv, lower milk cup equiv, and lower energy from solid fat and SoFAAS. Additionally, HLLF beef consumers had higher total HEI scores than LLHF beef consumers (51.0 ± 0.3 vs. 45.2 ± 0.3). Improved nutrient intake and diet quality were associated with consumption of lean beef. Support: USDA and Beef Checkoff
Cinnamon extract lowers blood glucose in hyperglycemic subjects
Authors: Stoecker Barbara J , Zhan Zhiwei , Luo Rencai , Mu Xiaoqun , Guo Xiuhua , Liu Yingying , Guo Qing , Zhou Jin , Kong Jiang , Zhou Zhenli , Cui Baoyin , Anderson Richard A
Published: June 2021

Based on insulin potentiating activity of cinnamon, effects of a water extract of cinnamon were tested in a 2‐mo double‐blind placebo trial with 137 participants in China. Mean±SEM age was 61.3 ± 0.8 years, BMI was 25.3 ± 0.3 and M/F ratio was 65/72. A placebo capsule or a 250 mg dried water‐extract cinnamon (CinSulin) capsule was given twice per day. At baseline, homeostasis model assessment‐estimated insulin resistance (HOMA‐IR) was significantly correlated with diastolic blood pressure (r=0.23) postprandial glucose (r=0.45) and insulin (r=0.42), triglycerides (r=0.29), fructosamine (0.23), BMI (r=0.29) and negatively correlated with HDLC (r=0.37). After 2 mo, fasting glucose decreased (p<0.001) in the aqua‐cinnamon‐supplemented group (8.85 ± 0.36 to 8.19 ± 0.29 mmol/L) compared with 8.57 ± 0.32 to 8.44 ± 0.34 mmol/L in the placebo group (p=0.45). Glucose 2 h after a 75 g carbohydrate load also decreased (p<0.0001) with CinSulin (15.09 ± 0.57 to 13.30 ± 0.55 mmol/L) compared to 14.18 ± 0.60 to 13.74 ± 0.58 mmol/L with placebo. Insulin concentrations and HOMA‐IR tended to be improved by aqua‐cinnamon supplements but differences were not significant. In summary, supplementation of a water extract of cinnamon had beneficial effects in subjects with hyperglycemia. (Supported by USDA‐ARS, Tang‐An Medical & Oklahoma State Univ).
Inhibition of Monkeypox virus replication by RNAi
Authors: Alkhalil Abdulnaser , Strand Sarah , Mucker Eric , Huggins John W , Jahrling Peter B , Ibrahim Sofi M
Published: June 2021

Orthopoxviruses cause human diseases with various severities and outcome ranging from mild conditions to death in fulminating cases. Currently, vaccination is the only protective measure against infection with these viruses and no licensed antiviral drug therapy is available. We investigated the potential of RNA interference (RNAi) in inhibiting Monkeypox virus (MPV) replication. Based on genome‐wide expression study, we selected 12 viral genes for targeting by small interference RNA (siRNA). Forty‐eight siRNA constructs were developed and evaluated in vitro for their ability to inhibit viral replication. Two genes, each targeted with four different siRNA constructs in one pool, were detrimental to viral replication. Seven siRNA constructs from these two pools, targeting either an essential gene for viral replication (A6R) or an important gene in viral entry (E8L), inhibited viral replication in cell culture by 65–95% with no apparent cytotoxicity. Further analysis using wild‐type and recombinant MPV expressing green fluorescence protein identified the most potent construct (siA6‐a) that inhibited virus replication with IC50 less than 5nM. These results identify A6R as an essential gene in MPV replication, and validate RNAi applications in developing antivirals for MPV and other Orthopoxviruses.

This work was supported by a grant from the DTRA, Project number 4.10022_07_RD_B.
Confirmatory factor analysis in nutrition and obesity research: Beyond dietary patterns
Authors: Lemacks Jennifer Lynn , Ilich Ernst Jasminka Z.
Published: June 2021

Confirmatory factor analysis (CFA) is primarily used for determining population's dietary patterns in nutritional epidemiological studies. However, CFA also allows substitution of easily attained variables (BMI, girth circumferences) to represent a variable otherwise measured with expensive equipment by certified staff (e.g. DXA). Our objectives were to: 1. Provide example of a model for adiposity determination in 189 postmenopausal, overweight women utilizing CFA; 2. Determine the strength of association between latent construct “adiposity” and total body fat percent (%fat) measured by iDXA as reference. Mplus Editor (v5) was used for the analysis with following: waist/hip circumference, BMI, %IBW. The construct was compared with %fat (iDXA). Model fit indices include chi‐square test (χ2), root mean square error or approximation (RMSEA), comparative fit index (CFI), and Tucker‐Lewis index (TLI). RMSEA/SRMR≥0.8 and CFI/TLI ≤0.90 or 0.95 indicates poor fit. Model fit indices (χ2=13.48, df=7, RMSEA=.10, SRMR=.03, CFI=.99, TLI=.98) were optimal and the latent construct “adiposity” significantly correlated with %fat (r=.20, p=.05). Each of the body adiposity indicators alone has certain limitations but based on this analysis, they may be more informative when used in combination as a defined latent construct and therefore substitute for less easily obtained variables.

USDA/CSREES/NRI#2004‐05287

Grant Funding Source: USDA
Maternal and breastmilk vitamin B12 correlated with infant status but was not influenced by maternal supplementation among HIV‐infected Malawian women in the Breastfeeding, Antiretrovirals and Nutrition Study (BAN)
Authors: Shahab‐Ferdows S , Allen LH , Hampel D , Bentley ME , Adair LS , Flax VL , Kayira D , Chasela CS , Kacheche Z , Tegha G , Kourtis AP , Jamieson DJ , van der Horst C
Published: June 2021

Vitamin B12 is important for infant growth and supplementation to lactating mothers could increase intakes to exclusively breastfed (EBF) infants in areas where maternal diet is poor in B12. HIV+ mothers in the Breastfeeding, Antiretrovirals, and Nutrition study were randomized to receive a lipid‐based nutrient supplement (LNS), containing the RDA for B12 (2.6μg), or no LNS. We examined the relationships among maternal and infant plasma (MP, IP) and breastmilk (BM) concentrations of B12 in a sub‐sample (n=521) at 2 or 6 wk (baseline) and 24 wk. At baseline MP B12 was positively correlated with BM (r=0.36) and IP (r=0.42), and IP and BM B12 were correlated (r=0.32) (all p<0.001). At 24 wk the relationships remained significant: MP B12 with BM (r=0.42) and IP (r=0.32), and IP with BM (r=0.29). At baseline there were no differences between LNS and no LNS in MP, BM or IP B12 concentrations. At 24 wk MP and BM B12 were higher in the LNS vs. no LNS group (p<0.005).However IP B12 was not increased by LNS despite strong correlation among IP, BM and MP. IP B12 is likely explained by maternal status in pregnancy. Maternal supplementation with 1 RDA of B12 was insufficient to improve IP concentrations through breast milk in this population, suggesting that higher levels of supplementation may be necessary.

Grant Funding Source: CDC (U48‐DP000059–01); Bill & Melinda Gates Foundation (OPP53107); Carolina Population Center (NICHD 5 R24 HD050924); USDA, ARS‐WHNRC.
Evaluation of physiological strain in law enforcement personnel wearing protective clothing and equipment
Authors: Yokota Miyo , Karis Anthony J , Goetz Victoria C , Tharion William J
Published: June 2021

Law Enforcement (LE) officers are frequently required to utilize personal protective equipment (PPE) to protect against potential exposure to chemical, biological, radiological or nuclear materials. However, thermal safety standards for LE‐PPE have not been established. This study assessed thermal‐work strain (TWS) levels of LE practitioners wearing PPE and carrying loads (~20 kg) during field training exercises. Ten male LE personnel involved in 1.5 hr training exercises including various activities (e.g., walking, running, climbing, carrying a human body) in cool environment (4.2 °C). Unobtrusively collected heart rate (HR) data integrated into a simple thermoregulatory model were utilized to evaluate core temperature (Tc). During a post‐study focus group, thermal sensation (TS) was assessed using a 9 point ordinary scale. Maximum HR ranged from 101 to 182 bpm and maximum predicted Tc ranged from 37.1 to 37.9 °C. Seven individuals reported transiently experiencing a TS ≥ 6 (= hot) during the training. The cool environmental conditions reduced TWS associated with PPE and vigorous physical activities (e.g., carrying/dragging a person). This approach should facilitate the establishment of LE‐specific standards for TWS and thermal injury prevention. This research was supported by NIJ and US Army NSRDEC. Authors’ views are not official views of US Army/DOD.
Catabolism of 4‐hydroxy‐2(E)‐nonenal (HNE) via omega oxidation in perfused rat livers
Authors: Berthiaume Jessica Marie , Li Qingling , Sadhukhan Sushabhan , Henry Fabrice , Tochtrop Gregory P , Brunengraber Henri , Zhang Guofang
Published: June 2021

Oxidative stress triggers peroxidation of polyunsaturated fatty acids (FA) to HNE. HNE detoxification is classically defined by conjugation reactions and excretion. However, we previously showed that HNE is also degraded through α/β FA oxidation by 2 parallel pathways via the carboxyl end of its metabolite, 4‐ hydroxynonanoate (HNA); a process which is inhibited by increased FA concentrations. In this study, other catabolic intermediates were identified in perfused hearts and livers by gas chromatography and mass spectrometry. Perfusion with [2H11]HNE or [3,413C2]HNA revealed intermediates (4‐hydroxynonanedioyl‐ CoA, 4‐hydroxyheptanedioyl‐CoA and 4‐hydroxypentanedioyl‐ CoA) in the liver indicative of ω oxidation, implying HNE catabolism can proceed from either end of the molecule. In hearts perfused with labeled HNE or HNA catabolism by α/β FA oxidation was evident, however, ω oxidation of HNE was not detected. The lack of HNE ω oxidation in the heart could play a role in its susceptibility to oxidant stress, particularly in pathological conditions such as diabetes and heart failure where alterations in FA metabolism and oxidative damage occur. These alternative detoxification pathways not only expand the understanding of how this endogenous toxicant is handled in the cell, but also suggest a novel interplay between energy metabolism and cellular antioxidant defenses. (Supported by NIH, NSF, AHA & CWRU)
Egg intake increases peripheral blood mononuclear cell expression of ATP‐binding cassette transporter A1 in parallel with toll‐like receptor 4 as a potential mechanism to reduce cellular inflammation in metabolic syndrome
Authors: Andersen Catherine J. , Blesso Christopher N. , Lee Jiyoung , Fernandez Maria Luz
Published: June 2021

We investigated the effects of excess dietary cholesterol from eggs on peripheral blood mononuclear cell (PBMC) expression of inflammatory mediators and HDL‐associated lipid transporters in metabolic syndrome (MetS). Thirty‐seven men and women consumed either 3 eggs (EGG, additional 534 mg of dietary cholesterol) or the equivalent amount of a cholesterol‐free egg substitute (SUB, 0 additional mg of cholesterol) in combination with a carbohydrate‐restricted diet for 12 weeks. All participants reduced body weight an average of 4.0 ¡Ó 2.8% with no difference between groups. While PBMC mRNA expression of tumor necrosis factor ƒÑ (TNF‐ƒÑ), interleukin 6 (IL‐6), and interleukin 1ƒÒ (IL‐1ƒÒ) did not change, toll‐like receptor 4 (TLR4) increased in EGG only (+75.3% vs. −6.2% in SUB). Changes in TLR4 expression were positively correlated with nuclear factor £eB (NF‐£eB) p65 DNA binding activity. ATP‐binding cassette transporter A1 (ABCA1) expression increased in EGG only (+127.0% vs. +31.5% in SUB) while ABCG1 did not change. Interestingly, changes in ABCA1 were positively correlated with changes in IL‐6, IL‐1ƒÒ, and TLR4 in EGG only, as were changes in ABCG1 and IL‐6 and IL‐1ƒÒ. These findings suggest that the increase in PBMC ABCA1 expression from egg intake is potentially due to an effort to reduce cellular inflammation in MetS. Supported by the American Egg Board and USDA Grant No. 2012–67011‐19914

Grant Funding Source: USDA Grant No. 2012–67011‐19914
Food Intakes Converted to Retail Commodities Database 2003–04
Authors: Bowman Shanthy , Martin Carrie , Carlson Jennifer , Clemens John , Moshfegh Alanna , Lin Biing‐Hwan
Published: June 2021

The objective was to develop the Food Intakes Converted to Retail Commodities Database 2003–04 (FICRCD) that converts foods in What We Eat In America (WWEIA), the dietary component of the National Health and Nutrition Examination Survey (NHANES) 2003–04. There are two major steps in the FICRCD development: (1) foods consumed in the surveys are separated into their ingredients and assigned to an appropriate retail commodity category, and (2) conversion factors are applied to adjust for food preparation and processing losses and convert foods consumed back to their retail commodity levels. There are eight major retail‐level commodity categories in the FICRCD: Dairy Products; Fats and Oils; Fruits; Grains; Meat, Poultry, Fish and Eggs; Nuts; Caloric Sweeteners; and Vegetables. Because each major category has several components, FICRCD has a total of 65 retail‐level commodities. In the WWEIA, NHANES 2003–04, tomatoes, potatoes, and leafy vegetables contributed 31, 29, and 6 percent, respectively to the Total Vegetables Commodity; oranges, apples, bananas and melons contributed 41, 20, 8, and 5 percent, respectively to the Total Fruit Commodity; and wheat flour, 75 percent to the Grains Commodity. FICRCD is useful for identifying disparities among socio‐economic groups in commodity consumption and consequently for nutrition and agricultural policy development. Funded by ARS and ERS, USDA
Effect of almonds on cardiovascular risk factors in Chinese patients with type 2 diabetes mellitus: study design (1035.1)
Authors: Chen Chiao‐Ming , Li Hung‐Ta , Liao Yen‐Yin , Liu Jen‐Fang , Chen C‐Y.
Published: June 2021

Almonds have shown to improve glucose control and lipid profile and ameliorate oxidative stress and inflammation in Chinese patients with type 2 diabetes mellitus (T2D). The main objective of this study is to further substantiate health benefits of average 60 g/d almond consumption on cardiovascular risk factors in the same population. We hypothesize that consumption of ~60 g/d almonds for 3 mo will improve glucoregulation, adipokine regulation, inflammation, endothelial function, lipid profile, and oxidative stress. We design a randomized, cross‐over, placebo controlled clinical trial with all meals being provided to all subjects. During the 2‐wk run‐in period, subjects will receive a control diet resembling typical Taiwanese diets, prepared based on the NCEP Step 2 guidelines. Subsequently, subjects will be randomized to receive either the control diet or the control diet with whole almonds incorporated to replace 20% calories. After a 2‐wk washout, subjects will receive the alternate diet for the other 3 mo. Biochemical biomarkers will include lipid profile, HbA1c, glucose, insulin, nitric oxide, e‐selectin, endothelial‐1, and intracellular adhesion molecule‐1, adiponectin, leptin, and resistin, hsCRP, IL‐6, IL‐10, retinol binding protein‐4, TNF‐α, oxidized LDL, and protein carbonyls in fasting serum, oral glucose tolerance test, brachial artery flow mediated dilation, and urinary isoprostanes.

Grant Funding Source: Supported by Almond Board of California and USDA
Increased expression of sodium transport proteins and Na,K‐ATPase activity in the outer medulla of kangaroo rat is related to its greater urine concentrating ability compared to Sprague‐Dawley rat (1137.7)
Authors: Aw Mun , Evans Kristen , Pannabecker Thomas
Published: June 2021

The kangaroo rat (Dipodomys merriami), a desert rodent, is able to concentrate its urine to more than 6,000 mosmol/kg water, nearly twice that of the SD rat. We hypothesize that active sodium reabsorption in the thick ascending limb (TAL) of the kangaroo rat outer medulla (OM) is greater than that of the SD rat, and plays a critical role in generating a steeper corticopapillary osmotic gradient. The OM was dissected and prepared for semi‐quantitative western blotting. We measured a four‐fold greater abundance of Na,K‐ATPase α‐1 subunit protein (NKA‐α1) (n=5, p<0.0001), and nearly two‐fold greater abundance of NKCC2 cotransporter and NHE3 protein expression in OM homogenates of kangaroo rat compared to SD rat (n=3, p<0.01). Three‐day water restriction in SD rat (water reduced to 40% of normal consumption) did not significantly affect NKA‐α1, NKCC2, and NHE3 expression (n=3, p>0.1). Na,K‐ATPase (NKA) activity was measured as the difference in ATP hydrolyzed in the absence and presence of ouabain, as determined by measuring release of inorganic phosphate. OM NKA activity in kangaroo rat was significantly higher than in control SD rat, 7778 ± 973 and 4886 ± 296 nmoles ATP hydrolyzed/mg protein/30 min respectively (n=3). However, the volume of mitochondria per unit inner stripe TAL cell volume is identical in both species. We conclude that the greater urine concentrating ability in kangaroo rat compared to the SD rat is associated with greater expression of active Na transport proteins and higher NKA activity, which increases sodium reabsorption in the TAL of the OM.

Grant Funding Source: APS, NSF IOS‐0952885 and NIH DK08338
The toxic effects of manganese on dopamine D2 receptor activation is not due to inactivation of the phospholipase C receptor signal transduction component (1143.7)
Authors: Rogers Keisha , Beaubrun Isaac , Catapane Edward , Carroll Margaret
Published: June 2021

Manganese (Mn), a neurotoxin causing Manganism a Parkinsons‐like disease, disrupts dopamine (DA) neurons, but the neurotoxic mechanism is not fully resolved. Reports postulate it more related to DA neuron dysfunction than degeneration. Lateral gill cilia of Crassostrea virginica are controlled by serotonergic‐dopaminergic innervations from their ganglia. DA causes cilio‐inhibition, serotonin (HT) cilio‐excitation. We showed post‐synaptic DA receptors in gill lateral cells are D2 type and Mn blocks effects of DA by blocking DA post‐synaptic receptors. The adenylyl cyclase component of the D2 signal transduction pathway was not effected by Mn. Phospholipase C (PLC), another component of the D2 signal transduction pathway, is inactivated by DA. We hypothesized PLC is not effected by Mn. We measured cilia beating using stroboscopic microscopy. Adding HT (10‐6 ‐ 10‐3M) to gill increased beating to 25 beats/min. Adding DA (10‐6 ‐ 10‐3M) decreased rates to 0. Mn (100 µM) prevented the cilio‐inhibitory response. m3M3FBS (10‐6 ‐ 10‐3M), a PLC activator, increased beating. U73122 (10‐6 ‐ 10‐4M), a PLC inhibitor, decreased beating. Mn (100 µM) had no effect on the actions of either drug. The study helps elucidate the neurotoxic mechanism of action of Mn by showing Mn disrupts dopamine’s actions at D2 post‐synaptic receptors, but not at the level of adenylyl cyclase or PLC in the signal transduction pathway.

Grant Funding Source: NIH‐ 2R25GM0600309, NSF‐ 0622197, NYSED‐ 0516041071
5‐Demethylnobiletin inhibits colon carcinogenesis in azoxymethane/dextran sulfate sodium‐treated mice (123.3)
Authors: Song Mingyue , Wu Xian , Zheng Jingkai , Xiao Hang
Published: June 2021

Our previous studies have demonstrated inhibitory effects of 5‐demethylnobiletin (5DN, a unique citrus flavonoid) in cultured colon cancer cells. Herein, we determined effects of 5DN on colitis‐associated colon carcinogenesis in azoxymethane/Dextran Sulfate Sodium ‐treated mice. Our results demonstrated that oral administration of 5DN significantly lowered colon tumor incidence by 54% and decreased overall tumor size by 49%. immunohistochemical analysis showed that 5DN had strong anti‐proliferative and pro‐apoptotic effects on colonic tumors. ELISA results demonstrated that 5DN suppressed the levels of IL‐1 and IL‐6 in colonic tissue by 89% and 87%. Immunoblot assay showed that 5DN significantly decreased the expression level of iNOS and COX‐2 in mice colon. HPLC analysis showed that oral administration of 5DN resulted in high levels of 5DN and its metabolites, i.e. 5,3'‐didemethylnobiletin, 5,4'‐didemethylnobiletin, and 5,3', 4’‐tridemethylnobiletin in the colonic tissues. Cell culture study showed that these metabolites, especially 5,3’‐didemethylnobiletin had much stronger inhibition on human colon cancer cells than 5DN. Flowcytometry analysis indicated that 5DN and its metabolites induced extensive cell cycle arrest and apoptosis. In conclusion, our results demonstrated that oral administration of 5DN significantly inhibited colitis‐associated colon carcinogenesis in mice and these effects can be partially attributed to the colonic metabolites of 5DN.

Grant Funding Source: This work was supported by funding from NIH, AICR and USDA.
Health professionals’ experiences providing care for obese women who breastfeed (131.2)
Authors: Garner Christine , Ratcliff Stephannie , Devine Carol , Thornburg Loralei , Rasmussen Kathleen
Published: June 2021

Obese women tend to stop breastfeeding (BF) earlier than normal‐weight women. Health professionals (HPs) are in a unique position to help obese women with BF, however, little is known about HPs’ strategies for assisting and experience providing BF care for obese women. To explore these issues, 34 HPs who care for pregnant or BF women were recruited in central New York. Qualitative interviews were audio‐recorded, transcribed and verified for accuracy; content analysis was used to identify themes. Across professions, HPs described providing care for obese women as requiring more time and physical work, and as being challenging. General strategies that HPs used included treating them the same as other women, using normalizing techniques, and treating them with dignity. The HPs disagreed about specific strategies to help obese women with BF including: discussing the benefits of weight loss, proper nursing positions and “making an air pocket” when putting baby to breast, and using nipple shields/shells. They agreed about emphasizing the benefits of BF for infant weight, that more could be done prenatally to prepare obese women for BF, and that home visits postpartum may benefit them. Providing care for obese women in a setting in which HPs feel like they don’t have enough time requires more resources. Comprehensive strategies are needed to provide consistent messages and assistance to help obese women breastfeed.

Grant Funding Source: Supported by grants from NIH (5T32HD007331) and USDA (Hatch 399449)
Sustaining dietary ketosis to improve blood flow and wound healing in young and aged Fisher rats (734.7)
Authors: Kesl Shannon , Jung Michelle , Prather Jamie , Sherwood Jacob , Gould Lisa , D'Agostino Dominic
Published: June 2021

Increasing evidence shows that limited energy and nutrient exchange is associated with age‐related impairment of wound healing. Ketone bodies, including β‐Hydroxybutyrate (βHB), are naturally occurring energy substrates, which may augment wound healing. We hypothesized that oral ketone supplementation without dietary restriction would enhance wound closure in young and aged Fisher rats by improving blood flow and supplying an alternative energy substrate. In our preliminary studies, we measured the magnitude and duration of ketosis following administration of a single 6.5g/kg dose of ketone precursors: R,S‐1,3‐Butanediol (KS1), Na+/K+ βHB salt and medium chain triglyceride (MCT) oil 1:1 mixture (KS2), or water in young and aged fisher 344 rats (n=6). Substances were administered through an intragastric gavage and whole blood samples (10 μl) were acquired for analysis of glucose and βHB at 0, 0.5, 1, 1.5, 2, 4, 8, and 12 hours following administration. Using an ischemic wound‐healing model, the ketogenic supplements were added to a standard diet fed ad libitum for 28 days. Laser Doppler imaging of the ischemic peri‐wound tissue every 7 days demonstrated significantly increased blood flow in young rats (n=10) fed KS1 at day 14 and 28 (p<0.001) and KS2 at day 7, 14, and 28 (p<0.01). In aged rats, blood flow was significantly increased in KS1‐fed at day 14 and KS2‐fed at days 7 and 14 (p<0.05). Wound size was significantly smaller in young rats fed KS1 and KS2 at 11 and 14 days following wound creation (p<0.05). In aged rats, KS1‐fed wounds were significantly smaller at days 11 and 14 (p<0.05) and in KS2‐fed at days 11, 14, and 28 (p<0.05). Wound healing improved by 3 days in aged KS1‐fed, 7 days in young KS2‐fed, and 10 days in aged KS2‐fed.

Grant Funding Source: Funded by the Office of Naval Research
Peptide bond cleavage through cyclization of asparagine (768.4)
Authors: Giaccone Zachary , Urbanski Laura , Siegart Nicolle , Reitter Julie , Mills Kenneth
Published: June 2021

Protein splicing is the post translational modification by which an internal peptide, or intein, separates itself from the surrounding residues, known as exteins. In the third step of the mechanism, asparagine cyclization, coupled to peptide bond cleavage, cleaves the intein from the neighboring exteins. We have studied this cyclization in two ways. The first is by a model peptide assay by which the cyclization of Asn is coupled to fluorescence. We observed that an increase of temperature correlates to a faster cyclization reaction, while a decrease in the dielectric of the solvent slows the cleavage of the peptide. The second way we studied cyclization is through the inteins from Trichodesmium erythraeum and Syncechococcus sp. PCC 6803. The intein from T. erythraeum ends in a Gln as opposed to the highly conserved Asn. By making mutations to the last residues, Asn or Gln, we revealed that T. erythraeum intein can complete C‐terminal cleavage with either amino acid, whereas Syncechococcus sp. PCC 6803 requires the asparagine in order to cleave. This supports the hypothesis that a naturally occurring cyclization with Gln could also be completed if the residue was mutated to the more kinetically favorable Asn. However, it is unlikely that a natural Asn cyclization would be able to react in the same way if mutated to Gln.

Grant Funding Source: Supported by the National Science Foundation under grant MCB‐1244089 and by the Dreyfus Foundation
Fresh pear consumption is associated with a better nutrient intake profile, better diet quality, and lower risk of obesity in adults (19+ y): NHANES (2001‐2010) (810.16)
Authors: O'Neil Carol , Nicklas Theresa , Fulgoni Victor
Published: June 2021

The association between fresh pear consumption with nutrient intake, dietary quality, and risk of obesity was examined in adults using data from NHANES (2001‐2010). Consumers (n=492) were defined by consumption of any amount of fresh pears using a 24‐hour diet recall. Means and ANOVA (covariate adjusted) were determined using appropriate sample weights. Logistic regression was used to determine odds ratios. The Healthy Eating Index‐2010 (HEI) was used to determine diet quality. Mean intake of fresh pears in consumers was 170±7.0 g/d. Intake of total sugars 133±3.0 v 123±0.8 g/d), fiber (23.9±0.6 v 16±0.1 g/d), vitamin C (105±6.6 v 87±1.3 mg/d), copper (1.6±0.1 v 1.3±0.01 mg/d), magnesium (333±11 v 293±1.5 mg/d), and potassium (3006±58 v 3722±12 mg/d) was higher (p<0.01) in fresh pear consumers than in non‐consumers. Intake of added sugars (16±0.6 v 19.2±0.2 tsp eq), total fat (75±1.3 v 82±0.3 g/d), MUFA (27.6±0.6 g/d v 30±0.1 g/d) and SFA (23±0.5 v 27±0.1 g/d) was lower (p<0.01) in fresh pear consumers than non‐consumers. The total HEI score was higher (p<0.01) (58.8±1.2 v 46.7±0.2) in pear consumers than in non‐consumers.The likelihood of being obese was 35% lower (p<0.01) in fresh pear consumers than non‐consumers. Fresh pear consumption was associated with better nutrient intake, diet quality and weight parameters in adults and consumption should be encouraged.

Grant Funding Source: Supported by Pear Bureau Northwest and USDA.
Biomechanics of Locomotion in Antarctic Sea Spiders (Pycnogonida)
Authors: Roberts Stephen , Mahon Andrew , Halanych Kenneth
Published: June 2021

Eight‐legged locomotion within chelicerates and other arthropod groups allows a diverse array of maneuvers (e.g. variable speeds; forwards, sideways and backwards walking; low turning radius turns) on uneven substrate. During straight path locomotion, an alternating tetrapod gait cycle (R1, R3, L2, L4 to L1, L3, R2, R4) is characteristic in terrestrial eight‐legged species. However, far less is known about octopod locomotion in aquatic or marine arthropods which, like their terrestrial relatives, face highly uneven and variable substrates, but also live in a highly resistant fluid medium. While aboard the RV Laurence M. Gould, we investigated the walking kinematics of Antarctic sea spiders (Pycnogonida) in the genera Pallenopsis, Colossendeis, Nymphon, and Austropallene collected from the Southern Ocean near the Antarctic Peninsula. In all groups, absolute and relative walking speeds were comparatively slow, with absolute walking speed ranging from 0.05 to 0.2 cm/sec, and relative walking speed ranging from 0.07 to 0.11 body lengths/sec. While footfall patterns sometimes approximated an alternating tetrapod gait, the gait pattern was variable and often non‐symmetrical, with leg pairs commonly transitioning from out of phase to in phase within the same individual. Extremely cold temperatures, low relative muscle mass, and a resistant fluid medium likely contribute to the extremely slow walking speeds of sea spiders, while their variable gait patterns compared to those of terrestrial spiders suggest that aquatic and terrestrial habitats impose different selective pressures on central pattern generators controlling leg movements and walking.

Support or Funding Information

A.R.M. and S.P.R. were supported by research grant ANT‐1043670 by the US National Science Foundation (NSF). K.M.H. was supported by research grant ANT‐1043745 by the NSF.