bluuebunny/crossref_metadata_2025 huggingface dataset:
Lafora disease: Convergence of neurodegeneration with plant starch metabolism
Authors: Gentry Matthew Shawn , Vander Kooi Craig W. , Taylor Adam O. , Pace Rachel , Meekins David A. , Guo Hou‐Fu
Published: June 2021

Reversible phosphorylation modulates many proteins involved in glycogenesis and glycogenolysis. Recent results from mammalian cells, mouse models, eukaryotic algae, and plants have identified a new phosphorylation target in glycogen metabolism, the glucan itself.

Mutations in EPM2A lead to Lafora disease, a fatal neurodegenerative disease. EPM2A encodes laforin, a bimodular protein with an amino‐terminal carbohydrate binding module and a carboxy‐terminal dual specificity phosphatase domain. We recently demonstrated that laforin removes phosphate from phosphoglucans.

Starch Excess 4 (SEX4) encodes a similar phosphatase in plants, and we demonstrated that laforin and SEX4 are the founding members of the glucan phosphatases. Multiple groups have now shown that these phosphatases dephosphorylate phosphoglucans, but the physical basis for the function of laforin and SEX4 is unclear.

We determined the crystal structure of SEX4, and found that it contains a phosphatase domain (DSP), carbohydrate binding module (CBM), and a previously unknown carboxy‐terminal domain. Hydrogen‐deuterium exchange mass spectrometry (DXMS) identified regions of the DSP domain that interact with glucans. Cumulatively, these results reveal the unique tertiary architecture of SEX4 that provides the physical basis for its function as a glucan phosphatase and present a model for the function of laforin.
Lipid dependent interaction of protein phosphatase 2A‐A (PP2A‐A) and G protein‐coupled receptor kinase 2 (GRK2)
Authors: Som Tapan , Stock Jeffry B
Published: June 2021

PP2A is a hetero‐trimeric Ser/Thr protein phosphatase. The A‐subunit serves as a scaffold protein onto which the catalytic subunit‐C binds at the carboxyl end and the B‐subunit at the amino‐terminal region. Using yeast 2‐hybrid selection, we found that GRK2, a Ser/Thr kinase, also binds to the A‐subunit. We present here evidence that in vitro using purified proteins GRK2 binds to PP2A‐A at its carboxyl terminal region. This binding is significantly enhanced in the presence of lipid. In Cos‐1 cells, while complex of GRK2/PP2A are absent in the cytosol, phospholipid, can generate such a complex from soluble components. In contrast, we show that such a complex exists in the membrane and associates with β2 adrenergic receptor (β2AR). Interestingly, this association occurs in the absence of an agonist while the presence of agonist does not cause a significant change in the level of their interaction. Whole cell experiments show that the phosphorylation of the receptor by GRK2 is increased upon agonist addition and further enhanced in the presence of an inactive phosphatase. These results suggest formation of GRK2/PP2A complex in the membrane potentially regulating the level of receptor phosphorylation.
Serum 25‐hydroxyvitamin D and physical function in older adults with knee osteoarthritis
Authors: Houston Denise K , Davis Cralen , Legault Claudine , Hausman Dorothy B , Nicklas Barbara J , Loeser Richard F , Kritchevsky Stephen B , Messier Stephen P
Published: June 2021

Vitamin D is necessary for optimal musculoskeletal function; however, many older adults are vitamin D insufficient putting them at risk of poor physical function. Baseline associations between serum 25‐hydroxyvitamin D (25[OH]D) and physical function were examined in older overweight and obese adults with knee osteoarthritis (OA) participating in the Intensive Diet and Exercise for Arthritis Study (n=420; mean age 65.6, 72% women, 17% black). At baseline, 17% of participants had 25(OH)D<50 nmol/L while 45% had 25(OH)D≥75 nmol/L. Serum 25(OH)D status was associated with 6 min walk distance and self‐reported physical function (SF‐36) using multi‐linear regression models adjusted for age, gender, race, education, and season (p≤0.05). Compared to participants with 25(OH)D≥75 nmol/L, those with 25(OH)D<50 nmol/L covered less distance on the 6 min walk (LS means ± SE: 458.8±9.2 vs. 477.9±7.1 meters; p=0.09) and had lower SF‐36 Physical Function scores (52.0±2.5 vs. 58.6±1.9; p=0.03). In summary, vitamin D deficiency was associated with lower physical function among older overweight and obese adults with knee OA. Supported by grants from NIH (R01 AR052528, P30 AG21332, M01 RR00211, and K01 AG030506).
Effect of Soy Protein on Blood Pressure in Prehypertensive/Stage 1 Hypertensive Adults
Authors: Lasrado Joanne , Alekel Lee D , Messina Mark , Heiberger Kevin , Dettmer Michelle , Franke Warren D
Published: June 2021

Research is mixed on the BP lowering effect of soy protein. However, even if such effects are modest, reductions in hypertension‐related mortality and morbidity in the population could be significant. Thus, the aims were to determine if daily intake of soy milk made from whole soy beans (WSB) or soy protein isolate (SPI) for 8 weeks reduces BP compared to cow's milk (CM) and if the nature of soymilk, WSB (Silk) or SPI (8th Continent) has a bearing on the degree of effect in pre‐/Stage I hypertensives. Participants (n=68) consumed the randomly assigned beverage equal to 21 g of protein/d in this single‐blind study. BP was measured in‐house using the recommended office assessment and with a 24‐hr ambulatory BP monitor at baseline, 4 and 8 weeks. Data analysis shows that while baseline BP did not differ across groups, responses to treatment differed based on the method used to assess BP. In‐house data showed significant improvements in systolic BP (SBP) for all groups and in diastolic BP (DBP) for only the CM group. Ambulatory data showed a significant improvement in DBP for only the CM group. Ambulatory BP likely provides more insight about overall BP because 1) measures are taken every 15 min for 24 h and 2) it also overcomes “whitecoat syndrome” (inflation of BP due to testing environment). Overall, soy protein did not have a greater effect in lowering BP compared to milk protein, and no difference between WSB and SPI was noted.
Baltimore Healthy Eating Zones program significantly improves food preparation methods among adult caregivers of low‐income African American youth
Authors: Yong Rachel , Rowan Megan T , Shin Ahyoung , Christiansen Karina , Coutinho Anastasia , Lwin Julia , Sharma Sangita , Martins Paula , Gittelsohn Joel
Published: June 2021

The Baltimore Healthy Eating Zones (BHEZ) study, conducted in low income Baltimore City corner stores and recreation centers, was a youth‐targeted multi‐level environmental intervention trial to promote healthy food consumption. The trial included increased availability of healthy foods and interactive health communication strategies. Impact was assessed on a sample of 83 of the 242 youth and their primary caregivers. Pre‐ and post‐intervention, caregivers were asked about their preparation methods for chicken, pork, ground beef, eggs, and greens. Scores were based on the degree to which the preparation method added or reduced fat content of the cooked food. Deep frying or pan frying was given a score of −1; baked, grilled, or steamed were given +1; pan‐fried, drained then rinsed was given +2. The change in pre‐ and post‐intervention score was calculated. The control group averaged a change in score of +1.76 ±1.86 and the intervention group averaged +2.31 ±2.02. Once this calculated score was stratified to < 2 or > 2, caregivers in the intervention group were found to significantly decrease the use of fat in cooking as compared to those in the control group (Pearson Chi‐Square, p‐value= 0.033). This preliminary finding suggests that BHEZ was effective in promoting healthier cooking methods among caregivers; thereby potentially improving the healthiness of foods provided to youths at home.

Grant Funding Source: The Robert Wood Johnson Foundation
The accuracy of the Figure Rating Scale in reflecting BMI among health professionals, limited‐income adults and children ages 6–11
Authors: Bagwell Rebecka , Wong Siew Sun
Published: June 2021

Objective

To determine the accuracy of figure rating scale (FRS) in reflecting the actual BMI either self‐reported or selected by parents for their children ages 6–11.

Method

50 health professionals (H) in nutrition and 35 parent‐and‐child pairs were surveyed in a 1‐time, individual, face‐to‐face interview. Demographic, monthly food expense, height, and weight data were collected. Self‐reported body size was collected using an adult FRS chart for adults, and a gender and ethnic‐specific child FRS chart for children. Correlations between self‐reported body size and actual BMI were calculated.

Results

Significant, positive correlations were found in HP (r=0.75), mothers (r=0.73), children (r=0.39 self‐reported vs. r=0.66 when mother selected a FRS for the child), normal BMI mothers (r=0.51), normal BMI children (r=0.66 when FRS selected by mother), obese children (r=1.0 self‐reported). HP with graduate degree(s) had the highest correlation (r=0.97) compared to HP with college degree and RD. Single mothers had higher correlation than mothers living with a spouse (r=0.98 vs. r=0.73).

Conclusion

Adult FRS was effective in estimating adult BMI but the child FRS was not effective in children ages 6–11. It is more accurate to measure the children's height and weight directly, or ask the mothers to report their child's body size.

This study was funded by USU Extension.

Grant Funding Source: USU Extension
Diet of male College Students Falls Short of American Heart Association Guidelines for cardiovascular disease risk reduction: Findings from Project NEW
Authors: Cessnun Kathryn , Baker Gavin , Lawson Samantha , Gilbertson Mark , Bersamin Andrea
Published: June 2021

Male college students represent an understudied group in nutrition research that may be at risk for poor diet quality. Project NEW (Nanooks Eating Well) was a study that aimed to assess the social determinants and quality of the diet of male college students (ages 18 to 25 years). Two 24‐hour dietary recalls were collected from 100 male college students using Nutrition Data Systems for Research Software. A web‐based questionnaire was also administered that assessed sociodemographic characteristics, food attitudes and beliefs, and cooking facilities. We found that the diets fell short of the current American Heart Association guidelines, with less than 20% of the population meeting the recommendations for fruit and vegetable servings, EPA/DHA fatty acids, dietary fiber, and saturated fat intake. Only 25% of the population met the guidelines for trans‐fat intake and 45% met the guidelines for cholesterol. About 35% of the population ranked eating not processed food as important and 45% ranked eating food prepared at home as important. These results suggest that the diets of college‐aged men may put them at risk for the development of cardiovascular disease in the future and this group may represent an important target group for a dietary intervention.
Lutein uptake is greater from egg yolks compared to nanoemulsions in Caco‐2 cells
Authors: Vishwanathan Rohini , Wilson Thomas , Nicolosi Robert
Published: June 2021

Lutein, an oxygenated carotenoid, plays an important role as macular pigment protecting the retina from blue light damage and oxidative stress. Unpublished findings showed significantly greater increases in plasma lutein after a one week intervention with egg yolks compared to lutein nanoemulsions in BALB/c mice. The goal of the present study was to compare digestive stability, micellarization efficiency and cellular uptake of lutein from egg yolks and 3 different nanoemulsions using in‐vitro digestion coupled with Caco‐2 cell uptake in the absence of any added fat. Lutein nanoemulsions are oil‐in‐water emulsions of lutein prepared using Microfluidizer® Processor technology that have a particle size of 150 nm. Phospholipon 85G (N1), Polysorbate 80 (N2) and Lipoid S45 (N3) were used to prepare 3 different nanoemulsions. Digestive stability of lutein from the egg yolks and the 3 nanoemulsions did not differ significantly. The efficiency of micellarization of lutein during the intestinal digestion was significantly greater from N1 and N2 compared to egg yolks (p <0.05). Cellular uptake of lutein micelles obtained from the egg yolks was significantly greater than N1, N2 and N3 (p <0.05). The egg yolk matrix was more suitable for lutein uptake by Caco‐2 cells despite the significantly smaller nanometer‐sized particles of the nanoemulsions. Supported by American Egg Board‐Egg Nutrition Center.
Serum antioxidant status and its association with metabolic syndrome among US adults: Findings from recent national data
Authors: Beydoun May A , Shroff Monal R , Chen Xiaoli , Beydoun Hind A , Wang Youfa , Zonderman Alan B
Published: June 2021

We examined the association between serum antioxidant status and the metabolic syndrome (MetS) using data from the National Health and Nutrition Examination Surveys (NHANES 2001–06) among adults aged 20–85 years. Sample sizes ranged between 3,008 and 9,099, depending on the outcome measure. MetS was defined with the National Cholesterol Education Program, Adult Treatment Panel III (NCEP ATP III) criteria. Elevated homeostatic model assessment insulin resistance (HOMA‐IR), C‐reactive protein (CRP) and hyperuricemia were also considered. Serum antioxidants included retinol, retinyl esters, carotenoids (α‐carotene, β‐carotene (cis+trans), β‐cryptoxanthin, lutein+zeaxanthin, total lycopene) and vitamin E and vitamin C. Multiple linear, zero‐inflated poisson and logistic regression models were conducted. MetS+ adults had consistently lower serum carotenoid concentrations compared to MetS− adults. Vitamin E was not significantly associated with MetS, while retinol+retinyl esters were inversely related to MetS in men only. Retinol+retinyl esters were also inversely related to elevated CRP and positively associated with hyperuricemia. Vitamin C was inversely related to MetS, HOMA‐IR and hyperuricemia. Serum antioxidant levels, carotenoids in particular, need be monitored to assess the effectiveness of MetS primary and secondary prevention related to lifestyle changes.
Contrast ultrasound imaging of the aorta alters vascular morphology and circulating von Willebrand Factor in hypercholesterolemic rabbits
Authors: Smith Brendon W , Simpson Douglas G , Sarwate Sandhya , O'Brien William D , Erdman John W.
Published: June 2021

Ultrasound contrast agents (UCAs) are intravenously infused microbubbles that add definition to ultrasonic images. UCAs hold clinical promise in diagnostic imaging of cardiovascular disease, but their biological effects are not known with confidence. We utilized a cholesterol‐fed rabbit model to evaluate these effects when used in conjunction with ultrasound to image the descending aorta. Male New Zealand White rabbits (n=41) were weaned onto an atherogenic diet containing 1% cholesterol, 10% fat and 0.11% magnesium. After 21 days, rabbits were exposed to contrast ultrasound at one of four pressure levels using either the UCA Definity or saline control (n=5 per group). Blood samples were collected and analyzed for lipids and von Willebrand Factor (vWF), a marker of endothelial function. Animals were euthanized at 42 days and tissues were collected for analysis. After adjustment for pre‐exposure vWF, high‐level ultrasound (2.3 or 3.5 MPa) resulted in significantly lower vWF one hour post‐exposure (p=0.017). This difference disappeared within 24 hours. Animals receiving UCA displayed reduced atheroma thickness in the descending aorta as compared to animals receiving saline, possibly due to ultrasonic disruption of the atheroma. Plaque disruption can lead to thrombosis and myocardial infarction. Further research is warranted to characterize the effects of this procedure. [Supported by NIH R37EB002641]
Seasonal differences in household food insecurity in Sidama Zone, southern Ethiopia
Authors: Joray Maya , G/Egziabher Tafere , Stoecker Barbara J , Hambidge K Michael
Published: June 2021

This study investigated seasonal changes in food insecurity in a rural community in Sidama southern Ethiopia. A longitudinal survey of 36 households was conducted in July of 2008 and January of 2009. Anthropometric and sociodemographic data as well as blood samples were collected along with administration of the Household Food Insecurity Access Scale (HFIAS). Body Mass Index (BMI) was not different by season. Although in both seasons the mean HFIAS category was overall “mildly food insecure”, the mean score in July was significantly higher than after the harvest in January (8.8 vs 3.6). In July, the season of greater food insecurity, zinc was lower (p <0.01) and albumin was related to the household's food storage (r = .40, p <0.02). The α1‐acid glycoprotein (AGP), a measure of chronic inflammation, was negatively associated with albumin (r = −.91, p <0.0001) and with the household's food storage (r = −.42, p <0.02). The husband's education also was correlated with the household food storage (r = .45, p <0.005). The HFIAS scores in both July and January were negatively associated (r = −0.36 and −0.48, respectively, p <0.03) with the household's ownership of enset plants, an indigenous drought‐resistant plant in southern Ethiopia. In this study, the HFIAS questionnaire captured seasonal variation in food supply; in addition several socio‐economic factors significantly influenced food insecurity (Supported by Hawassa University and Oklahoma State University and Swiss National Science Foundation).
Predicting energy expenditure from physical activity, heart rate and anthropometry in female Indian tea pluckers
Authors: Przybyszewski Eric , Hammons Julie , Venkatramanan Sudha , Haas Jere D
Published: June 2021

The objective of this study was to test a methodological procedure for estimating energy expenditure for a population of Indian female tea pluckers. Subjects (N=40; age=20–50 y) working on a tea estate in West Bengal, India participated in the study. Each subject wore an Actigraph accelerometer, Polar heart rate monitor and CosMed K4b2 metabolic analyzer during a 90‐minute period to assess minute‐by‐minute physical activity (PA), heart rate (HR) and energy expenditure (EE), respectively. The testing period was meant to replicate a normal tea picker's work day which included 2 periods of rest, 3 periods of picking while carrying weight (0, 5, and 10kg) and 3 periods of walking while carrying weight (15, 20 and 25kg). An EE prediction equation was generated using a branched method that first distinguishes times at rest from non‐rest (picking and walking) using accelerometer counts. Resting EE was estimated from age and BMI, while minute‐by‐minute non‐resting EE, was estimated from HR, age and BMI. Predicted EE will be used to evaluate the efficiency of performing work (weight of tea plucked/kcal EE) relative to iron status in an independent sample of 248 tea pluckers. We conclude that energy expenditure can be accurately predicted with a single branched equation based on PA, HR, age and BMI for a specific population participating in a known set of activities. Supported by the Mathile Institute and Micronutrient Initiative.
Household Food Insecurity and Dietary Intake among Women and Children in Rural Uganda
Authors: Garber Eden , Charles Dustin , Stefani Monique Centrone , Masinde Dorothy , Mazur Robert , Humphries Debbie
Published: June 2021

Household food security has been used to screen participants and evaluate program effectiveness for an agriculture and economic development project implemented by Volunteer Efforts for Development Concerns (VEDCO) in Kamuli District, eastern Uganda.

Objectives

This study was conducted to investigate dietary intake patterns and anthropometry as they relate to household food security among program participants.

Methods

Stratified random sampling was used to select 70 households with children under the age of 6 yrs within six geographic parishes, and dietary intake (24 hour recall) and anthropometry were measured on female caregivers (n = 70) and children under the age of six (n = 127).

Results

Prevalence of HAZ≤−2.0 in children was 28.6% and 29.5% of the caregivers were overweight (BMI≥25.0). Lower household food security and lower absolute wealth index were related to increased dietary fiber intake in the women and children (p<0.05), and decreased lipid intake in women (p<0.05) in bivariate analysis.

Conclusions

Changes in dietary quality with increased household food insecurity demonstrate household level strategies in the face of insecurity. These results will be used in the planning and evaluation of nutrition‐focused interventions in Kamuli and beyond. Research Support: The Wilbur G. Downs International Student Health Travel Fellowship and the Office of Student Research at the Yale School of Medicine.
Validity of Urinary Metabolites α‐CEHC and α‐CMBHC as Biomarkers of α‐Tocopherol Consumption: Correlations with Dietary and Plasma α Tocopherol
Authors: Lebold Katie M , Traber Maret G , Ang Alfonso , Chen Jasmine , Arab Lenore
Published: June 2021

There are no functional biomarkers of vitamin E status and plasma concentrations are a poor indicator of intake. Therefore, clarity on the relationship between α‐tocopherol (α‐T) intake with urinary vitamin E metabolites (α‐CEHC or α‐CMBHC) is needed to discern the utility of the latter as biomarkers in human studies. We examined in subjects (n=235) from Los Angeles, relationships between intake (via 24 h recall), lipid‐adjusted plasma levels, and urinary α‐CEHC or α‐CMBHC. In a subset who did not consume supplemental α‐T (n=202), Pearson correlation analysis of log‐transformed data showed stronger correlations between dietary α‐T and urinary α‐CEHC (r=0.38, p< .001) or α‐CMBHC (r = 0.34, p < 0.001), than between dietary and plasma α‐T (r = 0.21; p = 0.003) or between plasma α‐T and urinary α‐CEHC (r=.23, p < 0.001) or α‐CMBHC (r = 0.20; p < 0.05). In the subset of supplemental vitamin E users (n=33), the association of dietary α‐T and urinary α‐CEHC (r=0.44, p<0.01) was strong, but that of dietary α‐T with α‐CMBHC was not significant. These observations suggest that urinary levels of α‐CEHC might serve as a reliable biomarker of α‐T intake. Support provided by R01CA105048 and R01DK081761.
Autophagy and silica‐induced pulmonary fibrosis
Authors: Jin Yulan , Chen Shi , Yao Sanqiao , Fan Xueyun , Yuan Juxiang
Published: June 2021

The aim of this study was to investigate the potential role of autophagy in the pathogenesis of pulmonary fibrosis. Pulmonary fibrosis was induced by infusing 1 ml SiO2 (100 mg/ml) suspension to rat trachea. Rats were sacrificed 2, 7, 14, 21, and 28 days after the treatment to detect pulmonary fibrosis and autophagy. Autophagy was assessed by measuring the ratio of LC3II/LC3I, an established marker of autophagy, with Western blot and by detecting autophagic vacuoles with electron microscopy. Pulmonary inflammation and fibrosis were detected with H.E. staining. Pulmonary inflammation was evident early after the exposure to SiO2, which was followed by a marked increase in fibrosis. Compared to the control, LC3II/LC3I ratio and autophagosome in the experimental group were significantly increased 7 days after the treatment, indicating that silica can induce autophagy in the early stage of pneumoconiosis. However, the autophagic indexes were markedly lower in the experimental group than that of the control 14, 21, and 28 days after the treatment, implying that autophagy was suppressed in the late stage of pneumoconiosis. Taken together, these data suggest that autophagy may play different roles in the early and late stages of pneumoconiosis. These findings may provide important new insights into the mechanism underlying the pathogenesis of silica‐induced pulmonary fibrosis.
Strategies to enhance T cell reconstitution following severe immunodepletion
Authors: Martinez Mariela Raquel , Dudakov Jarrod , Young Lauren , van den Brink Marcel
Published: June 2021

Allogeneic hematopoietic stem cell transplant (AHSCT) is one of the only curative therapies for leukemia patients and others with hematological malignancies. Its use is restricted by major complications including post‐transplant immune deficiency which is devastating to the T cell compartment. Promotion of rapid and effective reconstitution of T cell immunity reduces incidence of transplant‐related complications and significantly improves the survival of HSCT recipients. Our previous studies show that precursor T cells (preT) generated ex vivo from stem cells on the OP9‐DL1 culture system can significantly enhance T cell reconstitution in AHSCT recipients. However, despite their regenerative benefits, the homing and seeding of preT to the thymus is relatively inefficient (only 1 out of every 10,000 iv injected preT engraft in the thymus). In the present study we examined strategies to improve the effectiveness of preT treatment. We found that direct administration of lineage‐depleted BM into the thymus can have significant benefit for T cell reconstitution. We also found that small changes in the purification strategy for seeding OP9‐DL1 cultures (based on differential CD150 expression) can have significant impacts on the generation and output of preT. These findings lay the foundation for improving T cell rejuvenation by enhancing the utility of preT as a potential clinical therapy.
Heat shock protein 70 can interact with and also prevent the high molecular weight FGF2‐induced apoptotic chromatin compaction
Authors: Ma Xin , Santiago Jon‐Jon , Nickel Barbara E. , Fandrich Robert R. , Kardami Elissavet
Published: June 2021

Fibroblast growth factor 2 (FGF2) is translated as high molecular weight (hi) and low molecular weight (lo) isoforms. We reported that increased nuclear expression of hi but not lo FGF2 elicited chromatin compaction and apoptotic cell death. We have begun investigating the role of hi FGF2 isoform‐specific interacting partners in modulating its pro‐apoptotic effect. Isoform‐specific affinity chromatography was used to capture interacting proteins from cell cytosolic and nuclear fractions. Nuclear proteins that bound only to the hi FGF2‐affinity column were identified by MS/MS peptide sequencing, and included the heat shock protein 70 (hsp70). Hsp70 was confirmed as a hi (but not lo) FGF2 interacting partner by co‐immunoprecipitation experiments. Overexpession of hsp70 prevented the hi FGF2‐induced chromatin compaction in cardiomyocytes. While hsp70 was mainly cytosolic in the absence of over‐expressed hi FGF2, it translocated to the nucleus in the presence of hi FGF2 over‐expression. The nuclear pattern of hsp70 localization partially overlapped with that of hi FGF2, suggesting interaction in situ. In conclusion, hsp70 interacts with hi but not lo FGF2, and is capable of preventing nuclear compaction by hi FGF2. It is possible that an in situ interaction between hsp70 and hi FGF2 is preventing the latter from exerting its apoptotic chromatin compaction effects.
Physical models of bacteriophage lytic enzymes: Sidwell Friends School S.M.A.R.T. Team Project
Authors: Hata Tommie , Nelson Daniel C , Herman Tim , Esch Tom , Estes Peter , Fishbein Daniela , Hamilton Kate , Katsnelson Alenka , Pyne Ottara , Robin Ari , Sako Angela , Stevens Eric , Stewart Ryan , Thomas Harper , Williams Lukas , Young Giorgio
Published: June 2021

The Sidwell Friends School S.M.A.R.T. Team (Students Modeling A Research Topic) has been working with the Nelson laboratory at the University of Maryland to design and produce accurate, three‐dimensional physical models of bacteriophage lysins using rapid prototyping (RP) technology. Lysins are bacterial cell wall hydrolases used by bacteriophage during progeny release. These enzymes in their pure form can be applied directly to Gram positive bacteria to quickly induce hypotonic lysis from without. This rapid activity shows promise as a novel antibacterial agent to both prevent and treat infections. Discussions with members of the Nelson laboratory allowed the students to design models of lysins to highlight their structural and functional characteristics. These designs were used to direct RP machines to build physical models of these enzymes. These physical models serve as “communication tools” used to enhance the understanding of lysins and their applications among the scientific and academic community. By contributing this tool to the Nelson laboratory research team, the students have the unique opportunity to experience and participate in the activities of a research laboratory. This work is supported by a grant awarded to Tim Herman by the NIH NCRR SEPA program and the HHMI Precollege Science Education Program.
Structural modeling of human DHFR in different ligand bound states
Authors: May James , Reed Colin , Bargman Jerrod , Huettig Emma , Chowdhury Parag , Bhabha Gira
Published: June 2021

Understanding the functional differences between bacterial and human enzymes is critical for the development of novel antibiotics to specifically target bacterial pathogens. Dihydrofolate Reductase (DHFR) is a ubiquitous enzyme present in both bacteria and humans that reduces dihydrofolate to tetrahydrofolate, which is a precursor for thymidylate synthesis, and is therefore essential for cell growth. For this reason, DHFR has been a focus in anticancer and antibiotic drug development. Crystollographic and NMR studies have elucidated how the E. coli human enzymes work. The E. coli enzyme undergoes large conformational changes in the active site loops as it progresses through the catalytic cycle. In contrast, the active site loops in the human enzyme are more rigid, and the enzyme undergoes a twisting hinge motion to accommodate ligand flux. We compared two human DHFR (hDHFR) crystal structures; one with hDHFR bound to NADPH, and the other model showing hDHFR bound to both the folic acid and NADP+. hDHFR:NADPH shows the active site with the hinge open. hDHFR:NADP+:folic acid shows the enzyme with the hinge closed, and the active site prepared for catalysis. Using 3D printing technology, we modeled these two conformations of hDHFR. These mechanistic differences between bacterial and human DHFRs can be utilized to design more specifically targeted antibiotics. Supported by a grant from HHMI Pre‐College Program.
A human milk oligosaccharide (HMO) influences cell proliferation in transformed human intestinal cells (CaCo‐2Bbe & HT‐29)
Authors: Holscher Hannah D , Davis Steven R. , Tappenden Kelly A.
Published: June 2021

This study aimed to determine the time course of impact of a model HMO on proliferation and differentiation in HT‐29 and CaCo‐2Bbe cells. Pre‐confluent HT‐29, pre‐confluent Caco‐2Bbe, confluent Caco‐2Bbe, and post‐confluent Caco‐2Bbe cells were used to model various stages of cell kinetics and differentiation along the crypt‐villus axis. Cells were randomized to 1 of 5 treatments: HMO at 0, 100, 200 or 400 mg/L or control (carbohydrate) for 24, 48 and 72 hrs. Dependent variables were cell proliferation via BrdU and cellular differentiation via alkaline phosphatase (AP). Compared to control, HMO decreased cell proliferation in pre‐confluent CaCo‐2Bbe at 72 hrs (p< 0.01) and pre‐confluent HT‐29 cells at 48 hrs (p<0.05) and 72 hrs (p<0.01). AP activity increased over time of incubation and was highest in post‐confluent CaCo‐2Bbe cells (p<0.01). AP activity was not significantly different among the treatment groups at any time point. A 72 hr incubation time was identified as the most appropriate to measure treatment effects on cell proliferation and differentiation. The HMO decreases cell proliferation in pre‐confluent states but the impact on cell differentiation is less clear. These results demonstrate that HMO influences growth related characteristics in transformed human intestinal cells, which suggests that HMO may have a role in promoting maturation of the neonatal gut.

Research funded by Abbott Nutrition.
Dietary protein upregulates fractional intestinal iron absorption and directly increases DMT1 transcript and protein expression
Authors: Cucchi Carrie Elizabeth , Gaffney‐Stomberg Erin , Sun Ben‐hua , O'Brien Kimberly , Kerstetter Jane , Insogna Karl
Published: June 2021

Iron (Fe) deficiency is endemic worldwide. We have recently found that increasing dietary protein dose‐dependently improves Fe balance in rats. This change is associated with increased DMT1 transcript expression, a key regulator of intestinocyte Fe uptake. To confirm the Fe balance results, Fe absorption studies were performed in rats fed either a 20% (control) or 40% (high) protein diet for 1 wk. Rats on the 40% protein diet absorbed 30±9% of ingested Fe while those on the 20% protein diet absorbed 18±8% (p= 0.005). To determine if DMT1 protein expression is also increased with increasing dietary protein, Western analyses were undertaken using duodenal mucosal lysates prepared from rats on the 20% and 40% protein diets. There was a trend towards higher DMT1 protein levels on the 40% diet (1.7±1.1 fold; p=0.46). To evaluate if amino acids could directly increase DMT1 transcript expression, Caco‐2 Bbe cells were exposed to either a mixture of amino acids or amino acid‐free media for 6 hrs. Amino acid treatment induced DMT1 mRNA by 1.6±0.3 fold as determined by qPCR (p<0.02). These data indicate that increasing dietary protein increases intestinal Fe absorption in part by upregulation of DMT1. Further, there appears to be a direct effect of amino acids on DMT1 transcript expression.

This work was supported by a HATCH grant from the Univ. of CT.
Common Consumer Questions Regarding Omega‐3s
Authors: Bangia Deepika , Palmer Debra M , Adler Audrey
Published: June 2021

Omega‐3 fatty acids (n‐3s) are gaining recognition in the scientific literature and the popular media. Research‐based, valid and reliable nutrition information on n‐3s is limited. Therefore a USDA eXtension website (ABC's of N‐3s) is being developed. This study was designed to assess consumer questions about n‐3s to inform the website development. Intercept interviews were conducted with the general public (N=352) in New Jersey's Newark Penn Station waiting rooms. Following the completion of a brief demographic survey, study participants were asked what they wanted to know about n‐3s. The participants' questions were answered from a list of responses that had been composed by a group of n‐3 researchers, unless a set response was unavailable. This process was repeated until participants' questions were exhausted. The most common questions asked were: What are n‐3s? How will n‐3s benefit me? Which foods are rich in n‐3s? What are other foods rich in n‐3s besides fish? Where can I buy foods rich in n‐3s? Are n‐3 rich foods more expensive? Do fish oil capsules have mercury in them? How much n‐3s should I consume? Are there any side effects to consuming n‐3s? What are some vegan and vegetarian sources of n‐3s? These results show that the general public has many questions regarding n‐3s, indicating a need for reliable resources on this subject. Research funded by the Center for Advanced Food Technology, Rutgers University.
Comparison of predictive equations for resting energy expenditure (REE) in obese and nonobese children and adolescents
Authors: Kim Myung‐Hee , Kim Eun‐Kyung
Published: June 2021

The aim of this study was to assess the accuracy of predictive equations for REE in obese and nonobese young people. Subjects were 137 healthy Korean students (51 obese, 86 nonobese) aged 7–18y. REE(MREE) was measured by indirect calorimetry. Predicted REE(PREE) was calculated using the 9 equations(Schofield‐W/WH, WHO‐W/WH, Henry‐W/WH, IOM, Maffeis, and Kim equation). MREE of obese students (1558±308 kcal/day) was significantly higher than nonobese (1232±238 kcal/day). The validity of the equations was evaluated on basis of prediction accuracy (the percentage of subjects predicted within 10% of the MREE), difference, RMSPE, % difference, and limits of agreement of Bland‐Altman method between PREE and MREE. The most accurate for nonobese students was the Maffeis equation (prediction accuracy 61%, difference 0±161 kcal/day, RMSPE 109 kcal/day, % difference 2±12%). Although the WHO‐WH equation was the most reliable in obese (difference 113±164 kcal/day, RMSPE 165 kcal/day, % difference 7±10%), the prediction accuracy of the equation was only 53% and noteworthy errors exist. Thus, this study suggests that a new obesity‐specific predictive equation should be derived to estimate REE for obese students accurately.

This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education, Science and Technology(2010‐0004472).
Dietary supplement use by US adult population in the 2007–2008 National Health and Nutrition Examination Survey
Authors: Luo Hanqi , Kennedy Eileen , Houser Robert
Published: June 2021

Data from the 2007–2008 National Health and Nutrition Examination Survey (NHANES) suggested that nearly half of the US adults age 20 to 69 reported taking at least one dietary supplement in the past month. The following factors were associated with a greater likelihood of supplement use: females, older adults, people with a higher level of education, non‐Supplemental Nutrition Assistance Program (SNAP) participants, and those in food secure households. In order to compare nutrient intakes between supplement users and non‐supplement users, daily intakes of eight nutrients were analyzed. When only considering nutrients from food, supplement users tended to consume greater amounts of Vitamin A, Vitamin C, Vitamin E, Folic acid, and Calcium; whereas, there was no association between supplement use and the remaining nutrients: Vitamin B‐12, Zinc and Iron. After including daily supplement use, supplement users consumed greater amounts of all eight nutrients. Therefore, supplement users were more likely to meet their Daily Reference Intakes (DRIs) both from only their food, and from their food and supplements. However, supplement users also experienced a higher risk of excessive nutrient intakes.
Nutrient biomarker patterns, cognitive function, and MRI measures of brain aging: a proof of principle study
Authors: Bowman Gene L. , Quinn Joseph F. , Shannon Jackilen , Dodge Hiroko , Frei Balz , Kaye Jeffrey , Howieson Diane
Published: June 2021

Clinical trials of single or few nutrients to slow the progression or delay the onset of Alzheimer's disease have been unsuccessful. One explanation may be related to more minimal effects that nutrients have in isolation. However, rigorous methods to define the ideal set of nutrients to inform this view have remained elusive. We applied a data driven method to generate distinct nutrient patterns derived from biological markers of diet and tested their relationship with cognitive and neuroimaging outcomes.

Thirty plasma nutrients were measured in 104 elders at risk for dementia. These nutrient biomarkers were selected on the basis of existing knowledge of association with neurodegeneration. Principle component analysis generated distinct nutrient biomarker patterns. Regression analysis tested cross sectional and longitudinal relationship with cognitive and MRI measures of brain aging.

Mean age was 87 (SD±10) and 62% were women. Eight nutrient biomarker patterns met out inclusion criteria (eigenvalue > 1.0). Three NBPs (NBP1, NBP5, NBP8) were associated with cognitive and neuroimaging outcomes in cross sectional analysis. Two NBPs, one favorable (NBP1) and another unfavorable (NBP8) explained significant variance in rates of change in Clinical Dementia Rating – sum of box score over a mean of 1.8 years.

Three distinct nutrient biomarker patterns were internally consistent. These may represent a successful formula of nutrition more capable of promoting cognitive health in people at high risk for Alzheimer's disease.
Protection of Oxidant‐induced astrocytes Injury by a Unique Cruciferous Nutraceutical
Authors: Jia Zhenquan , Zhu Hong , Li Yunbo , Misra Hara P.
Published: June 2021

The involvement of reactive oxygen species and electrophiles in the pathogenesis of various neurodegenerative disorders has provoked extensive studies on the use of exogenous antioxidative compounds to prevent oxidative neurodegenerative processes with little effects. In this study, we demonstrated that the cruciferous nutraceutical 3H‐1,2‐dithiole‐3‐thione (D3T) at micromolar concentrations (10–100 μM) has the potential to induce the levels of reduced glutathione (GSH) and NAD(P)H:quinone oxidoreductase 1 (NQO1), two crucial cellular defenses against oxidative and electrophilic stress in human astrocytes. In addition, pretreatment with D3T protected cells from oxidative and electrophilic neurocytotoxicity induced by various neurotoxicants including acrolein, 4‐hydroxy‐2‐nonenal (HNE), 6‐hydroxydopamine (6‐OHDA) and 1‐methyl‐4‐phenylpyridinium (MPP+). The results of this study may have important implications for the development of novel neuroprotective strategies against neurodegenerative disorders underlying Parkinson's disease.
Neuroprotective effect of Gastrodia elata Blume ethanol extracts (GEB) against beta‐amyloid‐induced BV2 cell death: Gastrodin and 4‐hydroxybenzaldehyde (4‐HBA) are the bio active candidates
Authors: Kim Sun‐Young , Back Hyang‐Im , Park Soo‐Hyun , Park Eun‐Ock , Oh Mi‐Ra , Kim Min‐Gul , So Byung‐Ok , Chae Soo‐Wan , Lee Geum‐Hwa , Kim Do‐Sung , Kim Hyung‐Ryong , Han Sang‐Young , Chae Han‐Jung , Chae Soo‐Wan
Published: June 2021

Increasing data from in vitro and in vivo studies indicate that amyloid beta‐induced damages in neurons and glia are mediated via ER (Endoplasmic Reticulum) stress. Gastrodia elate Blume ethanol extract (GEB) has been traditionally used as a folk medicine in Oriental countries. We investigated whether GEB, containing phenolic compounds, had a beneficial action on amyloid beta‐induced BV2 microglioma cell death and determined how this effect was produced. In addition, the bioactive candidates involved were identified. The treatment of amyloid beta showed BV2 cell death, dose and time dependently. Amyloid beta also induced ER stress proteins expression (CHOP, IRE‐1a, PERK and p‐eIF2a), dose and time dependently, except for GRP 78 protein. When cells were pre‐treated with GEB, Gastrodin or 4‐HBA, prior to amyloid beta BV2 cell death was inhibited. Furthermore, GEB, gastrodin or 4‐HBA regulated the cell death, and the expression of CHOP and GRP 78. Data from the study suggests that GEB, gastrodin, or 4‐HBA have protective effect on amyloid beta‐induced cytotoxicity in BV2 microglia cells. In conclusion, GEB especially gastrodin and 4‐HBA may provide therapeutic effects on Alzheimer's disease through attenuation of amyloid beta‐induced cell death.
Obesity increases carcinogenesis in a mouse model of basal‐like breast cancer (BLBC)
Authors: Sundaram Sneha , Freemerman Alex , Sampey Brante , Darr David , Bendt Katharine , Hua Kunjie , Troester Melissa , Makowski Liza
Published: June 2021

To determine whether diet‐induced obesity promotes post‐pregnancy carcinogenesis in a mouse model of BLBC. In this pilot study non‐parous C3‐Tag mice that are genetically prone to develop only BLBC were used. At 10 weeks old, baseline fat and lean mass were estimated using MRI imaging followed by initiation of diets. Mice, randomized into 3 groups, received low fat (10%; control) or high fat (45% or 60%) diets. Caloric intake was measured by determining the gram food intake/mouse/day. Mice were monitored for palpable tumors thrice weekly from the start of diet. Progression of diabetes and obesity was assessed every 2 weeks. Percent changes in fat mass were assessed monthly with MRI. Mice on all three diets did not exhibit any significant changes in blood glucose levels. Caloric intake was significantly higher on the 60% diet compared to the 10% and 45% diets. High fat diet (45% and 60%) fed mice exhibited higher fat mass, compared to mice on 10% diet after 3 months on diet. Diets did not alter tumor burden (tumor number) versus control diet. C3‐Tag mice exhibit an average tumor latency of approximately 18.5 weeks (n=469). Although, there were no differences observed in the 10% and 60% groups from the expected tumor onset, the 45% fat diet group exhibited a significant decrease in tumor onset by 2 weeks. Further analysis of potential diet‐induced alterations in tumor mass using 3‐D ultrasound is ongoing. In sum, an elevation in caloric intake and fat mass with increases in fat content of the diets is evident with no diabetic complications. We conclude that the 45% diet is sufficient to promote obesity and may decrease tumor latency in C3‐Tag mice. (U01 ES019472 NIEHS).
PROTEOMIC PROFILES OF DESMOID TUMORS AND OSTEOSARCOMA OF BONE AND SOMATIC SOFT TISSUE
Authors: RAO UMA N , CONRADS TOM P , HOOD BRIAN
Published: June 2021

DESMOID TUMORS AND HIGH GRADE OSTEOSARCOMAS OF BONE AND SOMATIC SOFT TISSUE HAVE DISTINCT PROTEOMIC PROFILES

Proteomic analysis of recurrent but non metastasizing desmoid tumors, and osteosarcoma (OS) with known metastatic potential occurring in older individuals was performed to identify potential predictive marker proteins for drug resistance and prognosis. Archived samples from nineteen desmoid tumors and fourteen primary osteosarcoma occurring in soft tissue, bone were collected following IRB approval. Five micron thick sections were obtained from paraffin blocks of tumor and scraped, digested and analyzed by mass spectrometry. Protein abundances determined by spectral counting and used for hierarchical cluster analysis. Increase in HSP70, clusterin, emelin, creatine kinase B and carbonic anhydrase noted predominantly in OS. Results were validated for myoglobin and clusterin by immunohistochemical stains.. Proteins at higher abundance in OS relate to oxidative stress and hypoxia. HSP90 and clusterin may be markers of aggressive extraosseous osteosarcoma and may be indicative of drug resistance. The absence of these proteins in desmoids support their being considered benign locally recurrent tumors.
Altered retinoid metabolism in breast cancer
Authors: Pingitore Attilio , Perri Mariarita , Cione Erika , Kane Maureen A.
Published: June 2021

Cancer, a disease of uncontrolled cell growth arising from aberrant cell proliferation, differentiation and apoptosis is the second leading cause of death in the United States with 26% of new cases attributed to breast cancer. The term “retinoids” describes compounds that include vitamin A (retinol), its metabolites, and synthetic analogs that exhibit vitamin A activity. Retinoic Acid (RA) is an active metabolite of vitamin A and is essential to a wide range of physiological processes, including cell proliferation, cell differentiation, and apoptosis. RA homeostasis is maintained through dietary intake of retinol, and its storage, mobilization, transport, and biosynthesis into RA. The biosynthesis and shuttling of RA to specific nuclear receptors (RAR, RXR, and/or PPARb/d) dictates the outcome of RA signaling. Defective RA metabolism in local populations of cells is thought to be a main initiator of mammary tumor formation and progression. With robust and rigorous analytical methods (LC‐MS/MS) we have studied retinol metabolism in a panel of breast cancer cells together with biochemical and genetic insights into the mechanisms involved in the disruption of vitamin A metabolism in mammary carcinoma.