bluuebunny/crossref_metadata_2025 huggingface dataset:
Does changing the PUFA content of a high saturated fat meal influence postprandial lipid and lipoprotein expression in children with nonalcoholic fatty liver disease?
Authors: Mager Diana , Rodriguez-Dimitrescu Carla , Yap Jason , Mazurak Vera , Vine Donna , Jetha Mary , Ball Geoff
Published: June 2021

Background

Hyperinsulinemia leading to altered lipid metabolism induced by high saturated intakes may contribute to NAFLD. We hypothesized that consumption of a ↑saturated/↓PUFA meal would evoke a postprandial metabolic environment characterized by prolonged hyperinsulinemia, hypertriglyceridemia and hypercholesterolemia when compared to a ↑saturated/↓ PUFA meal in childhood NAFLD.

Methods

We studied 36 children (n=15 NAFLD; n=21 lean controls) following consumption of two meals: ↑saturated/↓ PUFA vs↑saturated/↓PUFA that were equivalent except for PUFA content (2.8% vs.4.9%). Blood for analysis of total‐HDL‐LDL‐cholesterol, triglycerides (TG), glucose, insulin, non‐esterifiedfatty‐ acids (NEFA), Apo‐B48, ApoB‐100, ApoC‐III and ALT (fasting) was collected at 0, 1, 3 and 6 hr. Data was analyzed using the trapezoidal method for area under the curve (AUC)/iAUC.

Results

: Mean age and BMI‐z (±SD) of participants was 13.4±2.8 and 2.3±0.4 (NAFLD); and 13.8±2.6 and 0.1±1.0 (lean). Mean insulin AUC (371 ±238 vs. 82±26) and iAUC (210 ± 179 vs. 41 ±20) for NAFLD was significantly higher than for controls (p<0.05). No significant differences in AUC/iAUC for insulin, TG, total‐and‐LDL cholesterol or ApoB48 was observed between meal types (p>0.05), although significant differences in iAUC NEFA (1.27±0.91 (↓PUFA) vs. 0.42±0.17 (↑PUFA), HDL‐cholesterol and ApoB‐100 by meal type in children with NAFLD and lean controls were observed (p<0.05).

Conclusions

: Children with NAFLD experience postprandial hyperinsulinemia and marked changes in lipid, cholesterol, and lipoprotein expression when compared to lean children when fed meals with high saturated fat with varying PUFA content.
Targeting DNA repair protein as an approach for sensitizing cancer cells to methylseleninic acid
Authors: Tzeng Tiffany J , Wu Ryan TY , Rocourt Caroline RB , Cheng Wen-Hsing
Published: June 2021

DNA‐dependent protein kinase (DNA‐PKcs) is part of the DNA‐PK complex that senses and repairs DNA double strand breaks by the nonhomologous end‐joining pathway. The WRN protein is a member of the RecQ DNA helicases and plays essential roles in the maintenance of genome stability. Whereas mutations in WRN result in Werner syndrome that displays premature aging phenotypes and cancer predisposition, the only human PRKDC (DNA‐PKcs) mutation known to date is the L3062R missense mutation found in a severe combined immunodeficiency patient. To test the hypothesis that targeting DNA repair proteins can improve the cytotoxic effect of selenium in cancer cells, we tested the sensitivity of 1) WRN small hairpin RNA (shRNA) and scrambled shRNA U‐2 OS osteosarcoma cells and 2) DNA‐PK−/− and control HCT 116 colorectal cancer cells to methylseleninic acid (MSeA, 0–5 μM). Here we show that DNA‐PKcs or WRN deficiency sensitizes the cancerous cells to MSeA‐induced cytotoxicity in a dose‐dependent manner. DNA‐PKcs or WRN does not affect MSeA‐induced ATM phosphorylation on Ser‐1981 or H2A.X phosphorylation on Ser‐139. However, WRN promotes recovery of the cells from MSeA‐induced DNA damage. Taken together, DNA‐PKcs and WRN protect cancer cells against MSeA‐induced cytotoxicity, suggesting that targeting DNA repair pathway is a promising approach for improving selenium chemoprevention.
Selenoprotein induced M1 to M2 murine macrophage phenotype switching is imperative in helminth parasite clearance
Authors: Nelson Shakira M , Carlson Bradley , Urban Joseph , Lei Xingen , Hatfield Dolph , Prabhu K. Sandeep
Published: June 2021

Selenium (Se) in the form of selenoproteins, imparts many beneficial health and anti‐inflammatory properties. In the present study, we investigated the anti‐inflammatory activity of Se using a LPS and IL4 treated murine BMDM model. IL4‐treated macrophages supplemented with 100 nM Se significantly increased the expression of alternatively activated macrophage (M2) markers, Arg‐I and Mrc‐1. Se treatment also increased the enzymatic activity of Arg‐I and surface expression of Mrc‐1. Conversely, expression of classically activated macrophage (M1) markers, TNFα, and IL1β, were significantly decreased in LPS treated macrophages cultured in 100 nM Se and IL4, suggesting a synergistic effect of Se and IL4. Additional studies utilizing tRNA[Ser]Sec (Trsp) KO mice macrophages, characterized by a total loss of selenoprotein expression, showed a complete abrogation of Arg‐I activity, even under high Se (250nM) conditions, demonstrating the importance of selenoproteins. Current in vivo studies, using the helminthic parasite Nippostrongylus brasiliensis, show a significant increase in intestinal M2 marker expression, in addition to a decreased presence of intestinal adult worms and fecal eggs, in Se supplemented mice. These data suggest that optimal selenium status, in the form of selenoproteins, is critical to skew macrophage activation to promote resolution of (parasite) infection. NIH grant DK077152.
Folic Acid Supplementation Alters Adipose Tissue Lipid Metabolism
Authors: Kelly Karen B , Proctor Spencer D , Field Catherine J , Jacobs Rene L
Published: June 2021

Mandatory folate fortification polices in Canada and the United States has resulted in high levels of circulating folate in a large proportion of the population. At the same time, a trend toward increased energy intake and reduced energy expenditure has lead to an increased prevalence of obesity, characterized by an accumulation of TAG in adipose tissue. Folate plays an essential role in maintaining S‐adenosylmethionine (AdoMet), an important biological methyl‐donor. AdoMet functions in the formation of phosphatidylcholine from phosphatidylethanolamine catalyzed by the enzyme phosphatidylethanolamine N‐methyltransferase (PEMT). PEMT activity contributes to storage of triacylglycerol (TAG) in adipocytes.

The objective of this study was to investigate the effect of dietary folic acid supplementation on adipose tissue lipid metabolism. Sprague Dawley rats were fed a high fat diet containing 0.75 mg or 7.5 mg folic acid for 12 weeks.

Adipose tissue weight was significantly greater in rats fed excess folic acid. This was accompanied by an increased expression of genes involved in re‐esterification of fatty acids to glycerol, suggesting elevated TAG formation. Up‐regulation of genes involved in phospholipid biosynthesis supports increased storage of TAG in adipocytes upon folate supplementation.

In summary, folate supplementation enhances TAG storage in adipose tissue of high fat diet‐fed rats.

Grant Funding Source : CIHR, Food and Health Innovation Initiative
Dietary vitamin D3 at 50x the adequate intake increases apoptosis in the quadriceps of the female G93A mouse model of amyotrophic lateral sclerosis: a pilot study
Authors: Parkhomenko Egor , Milionis Angelica , Gianforcaro Alexandro , Solomon Jesse Adam , Hamadeh Mazen Jamal
Published: June 2021

Background

We previously showed that dietary vitamin D3 (D3) at 50x the adequate intake (AI) increased functional capacity in the transgenic G93A mouse model of amyotrophic lateral sclerosis (ALS), with females exhibiting signs of D3 toxicity. ALS is a fatal neuromuscular disease characterized by death of motor neurons in the CNS, muscle weakness and paralysis.

Objective

In this pilot study, we analyzed the quadriceps of G93A mice following dietary D3 supplementation at 50x the AI for contractile proteins (α‐actin; troponin C, trop C), inflammation (IL‐6), cell stress response (Hsp70), mitochondrial efficiency (UCP3) and apoptosis (CASP3 cleaved/pro ratio), mechanisms contributing to ALS pathology.

Methods

Starting at age 25 d, 41 G93A mice were provided food ad libitum with either adequate (AI; 1 IU D3/g feed; 12 M, 11 F) or high (HiD; 50 IU D3/g feed; 10 M, 8 F) D3. At age 113 d, the quadriceps were analyzed for protein content. Differences were considered significant at P ≤ 0.10.

Results

In females, HiD had 87% higher CASP3 vs AI (P = 0.084). Males had 59% higher α‐actin (P = 0.063), 21% lower trop C (P = 0.017), 42% lower IL‐6 (P = 0.099) and 58% higher Hsp70 vs females (P = 0.097).

Conclusion

Dietary D3 at 50x the AI increases apoptosis in the quadriceps of female G93A mice, which could be due to D3 toxicity. (Supported by NSERC and Faculty of Health‐York U).

Grant Funding Source : Supported by NSERC and Faculty of Health‐York U
Increased dietary saturated fat intake is negatively associated with compliance to exercise regimen and improvement in fitness parameters
Authors: Krishnan Sridevi , Gustafson Mary , Campbell Caitlin , Souza Elaine , Keim Nancy L
Published: June 2021

Objective

To determine if dietary intake of fat, sugar, and/or sodium affects improvement in aerobic fitness in response to a 6‐mo endurance and resistance training program in peri‐menopausal women.

Methods

18 women aged 42–52y underwent a 6 mo exercise intervention, working out 6 d/wk, 60 min/d. The program included daily aerobic activity at ≥ 50% of heart rate (HR) reserve and resistance exercises 3 d/wk. Monitors recorded heart rate during exercise, and compliance was calculated using the number of minutes above prescribed HR. Improvement in aerobic capacity was measured by VO2max at beginning and end of the study. Dietary intake was assessed by 24h recalls on 4 d in 1stmonth and 4 d in 6thmonth of intervention.

Results

Total fat and saturated fat intake were negatively correlated to compliance (r = −0.501, p<0.05; r = −0.511, p<0.05), but added sugars, total sugars and sodium intake were not. Change in VO2max was positively correlated with compliance (r = 0.475 p<0.05), but negatively associated with saturated fat intake (r= −0.416, p<0.05).

Conclusion

Our observations on the association of fat intake, exercise compliance, and fitness improvement warrant further investigation to understand their underlying biological and behavioral significance. Funded by USDA CRIS 5306 51530 019 00D

Grant Funding Source : USDA
Preventing excessive weight gain by encouraging healthy eating habits among adolescents in Brazil: a randomised community trial
Authors: Cunha Diana , Souza Bárbara , Pereira Rosangela , Sichieri Rosely
Published: June 2021

Objective

Effect of an educational programme aimed to discouraging sweetened beverages and cookies, crackers and chips and encouraging fruits consumption, to prevent excessive weight gain.

Methods

20 classes were randomly assigned in to intervention or control. From 572 students enrolled, 399 students participated to the study (intervention: 10 classes with 207 individuals; control: 10 classes with 192 participants). There were nine sessions with nutrition education in the intervention group during the school year 2010. For all interventions, parents and teachers were included and received specific materials. The main outcome was the change in BMI. Intention‐to‐treat analysis was performed taking into account the cluster (classes) effect.

Results

BMI change was not significantly different between groups (p=0.22), however, consumption of sweetened beverages (mean difference= −0,20; 95% CI=−0,30; −0,11) and cookies (−1,35; 95% CI = −0,20; −0,75) showed a greater reduction and fruits consumption showed a greater incrase (0,17; 95% CI =0,01;0,34) in the intervention compared to control.

Conclusion

The incentive to adopt healthy eating habits did not lead to changes in BMI, but changed dietary habits. Strategies based exclusively on the quality of diet is not enough to reduce weight gain.

Trial Registration: clinicaltrials.gov. Identifier: NCT01046474.

Funding: PAHEF, FAPERJ, CNPq.

Grant Funding Source: Pan American Health and Education and Brazilian Research Council
Beverage choices and contributions to food and nutrient intakes differ by smoking status: Results from What We Eat in America (WWEIA), NHANES 2005–2008
Authors: Sebastian Rhonda , Enns Cecilia Wilkinson , Goldman Joseph , Zizza Claire
Published: June 2021

This study sought to identify relationships between adults’ smoking status and beverage intake patterns. A nationally representative sample of men and women 20+ years (n=9495) with a complete 24‐hour dietary recall and smoking status data in What We Eat in America (WWEIA), NHANES 2005–2008 was analyzed. Respondents were classified as current (n= 2148), former (n=2430), or never smokers (n= 4917). Regression procedures provided adjusted estimates by smoking status of beverage group intakes, likelihood of reporting each beverage type, and contributions of beverages to nutrient and food group intakes. Relative to former and never smokers, current smokers have higher mean intakes of total beverages, coffee, sugary drinks, and beer (p<.001) and are more likely to report coffee and beer and less likely to report plain water (though mean daily water intakes do not differ). Total daily energy intake does not vary by smoking status, but beverages provide more energy (mostly “empty calories”), carbohydrate (mostly added sugars), and caffeine to current than to former and never smokers. Beverage choices of adults overall need improvement, and those of smokers are particularly poor. Funding source: ARS, USDA.
Beverage intakes by weight status ‐‐ do obese individuals drink more? Results from What We Eat in America (WWEIA), NHANES 2005–2008
Authors: Chester Deirdra Nicole , Sebastian Rhonda S. , Enns Cecilia Wilkinson , Goldman Joseph D.
Published: June 2021

This study sought to measure relationships between weight status and beverages intakes. A sample of adults 20 years of age and older with a complete 24‐hour dietary recall and weight status data in What We Eat in America (WWEIA), NHANES 2005–2008 was examined. Underreporters were identified using the Goldberg equation and excluded from further analyses. Individuals in the remaining sample were classified as normal weight (BMI>=18.5 and <25.0; n = 2194), overweight (BMI>=25.0 and <30.0; n=2515), or obese (BMI>=30.0; n=2206). Regression procedures provided adjusted estimates by weight status of beverage group intakes, likelihood of reporting each beverage type, and contributions of beverages to nutrient and food group intakes. Relative to normal weight men, overweight and obese men drank more total beverages and low‐calorie drinks (p<.001). Among women, the obese drank more low‐calorie drinks than their normal weight counterparts. Overall, few differences were found in contribution of beverages to food and nutrient intakes by weight status. Although obese individuals consume more beverages overall, they do not account for higher intakes of calories or nutrients. This study shows that regardless of weight status, all adults are in need of interventions aimed at educating them on healthy beverage choices. Funding source: ARS, USDA.
New roles of erythritol identified via transcriptomic profiling
Authors: Berger Alvin , de Cock Peter , den Hartog Gertjan , Boesten Daniëlle , Bast Aalt
Published: June 2021

Erythritol (ERT) is a polyol bulk sweetener with endothelial‐ and oxidant‐protective roles in diabetic rats and human umbilical vein endothelial cells (HUVECs). To unravel mechanism(s) besides radical scavenging, we studied herein transcriptional effects of ERT in HUVECs under high glucose (HG) conditions using Affymetrix microarrays. In response to ERT under HG (HGERT), 521 genes were changed vs HG (p<0.05; 0.94–1.13 fold change). Over‐represented canonical pathways included: TCA cycle/oxidative phosphorylation/electron transport chain; focal adhesion; insulin signaling; and the polyol pathway. Non‐canonical networks included PIK3R1, NFκB, HNF, XBP1, SOS, and RELA. These canonical and non‐canonical pathways are linked to diabetes onset, insulin signaling and production of adhesion molecules/nitric oxide. Many genes were oppositely regulated with HGERT/HG vs HG/NG (normal glucose) demonstrating protective effects of ERT. Notable changed genes were succinate dehydrogenase complex (SDHC‐down‐regulated), linked to energy production; and interferon α‐inducible protein (IFI6‐up‐regulated), linked to immune responses. In addition to its antioxidant activity, ERT influenced expression of various diabetes‐linked genes and pathways. ERT could be a clinically‐relevant nutritional‐ or pharmaceutical ingredient for prevention and treatment of diabetic complications or vascular damage.
Delta‐6 desaturase inhibition reverses cardiolipin remodeling and hypertrophy, but not mitochondrial dysfunction, in the aged mouse heart
Authors: Mulligan Christopher M , Le Catherine H , de Mooy Anthony Bret , Chicco Adam J
Published: June 2021

Alterations in the fatty acid composition of cardiolipin (CL), a mitochondrial phospholipid, may contribute to mitochondrial dysfunction associated with myocardial aging, but its mechanism and role in this process remains unclear. We hypothesized that CL remodeling in the aged heart, characterized by a loss of linoleic acid (LA) and increase in highly unsaturated fatty acids (HUFAs), results from increased activity of delta 6‐desaturase (D6D), which catalyzes the production of HUFAs from essential fatty acids such as LA. Aged (24 mo) C57Bl/6 mice were administered a selective D6D inhibitor (SC‐26196, 100 mpk/d; SC) or no drug for 4 weeks, followed by isolation of cardiac subsarcolemmal (SSM) and interfibrillar (IFM) mitochondria for assessment of respiratory function and CL composition. Aged hearts exhibited a classic pattern of CL remodeling in IFM, but not SSM, which paralleled a 25% loss in state 3 respiration in IFM only. SC reversed CL remodeling in IFM, but had no effect on mitochondrial respiration. Interestingly, SC also reversed left ventricular dilatation, reduced heart weight and attenuated contractile dysfunction in aged mice. These studies indicate that D6D plays a pivotal role in CL remodeling in the aged heart, but argue against the role of this process in mitochondrial dysfunction. The mechanism by which D6D influences cardiac remodeling and function warrants further investigation. Funding: AHA
Effects of flaxseed oil supplementation and FADS genotype on declarative memory abilities in toddlers
Authors: Cheatham Carol L
Published: June 2021

Objective

Docosahexaenoic acid (DHA) is integral to neuronal function and subsequent cognition. Biosynthesis of DHA from its precursor alpha‐linolenic acid (LNA) is controlled by the FADS gene complex. Previously unobserved genetic differences may be a confound in studies of biosynthesis. This intervention was designed to determine the cognitive effects of providing substrate (flaxseed oil) to a system with little exogenous DHA intake (human toddlers).

Methods

16‐month‐olds (n=65) were enrolled in a 4‐month double‐blind intervention and randomized to flaxseed or corn oil. Parents squeezed 2400 mg of oil into their toddlers’ food daily for 180 days. Memory was assessed at 0, 7, 180, and 187 days using a non‐verbal declarative memory paradigm. Toddlers were exposed to 4 new imitation events and were tested for memory immediately, after 20 min, and after 7 days at baseline and end (total events = 8). Genotype at rs174575 on the FADS2 gene was determined.

Results

Baseline, 20‐min delay, and 7‐day delay results indicate no differences in memory performance when analyses are based on participant genotype. However, when analyses are completed with maternal genotype, children of homozygous recessive mothers earned lower scores on the memory tasks.

Conclusion

Maternal genotype needs to be considered instead of or in addition to the participants’ genotypes when assessing the effects of fatty acid supplementation.
Consumption of foods that meet the American Heart Association Heart‐Check Program is associated with better diet quality and lower cardiovascular disease risk
Authors: Fulgoni Victor L , Carson JAS , Johnson RK , Kris-Etherton PM , Lichtenstein AH , Stitzel KF
Published: June 2021

The American Heart Association administers the Heart‐Check Program (HCP) which places a heart icon on food packages based on specific nutritional criteria. We evaluated the association of consuming foods that meet the HCP criteria with overall diet quality, food group/nutrient intakes, body mass index (BMI) and blood pressure using data from the National Health and Nutrition Examination Survey, 2003–2006. Adult (19+ yrs) participants were stratified into quartiles based on % energy consumed from HCP foods (≤6.3% energy in the lowest and ≥22.2% in the highest quartile). Regression analyses were conducted to determine the relationship between consuming HCP foods and diet quality (Healthy Eating Index), food group/nutrient intakes, and physiological parameters while logistic regression analyses were conducted to assess relationship with CVD risk factors. Diet quality was higher in consumers of HCP foods versus non‐consumers (41.0 ± 0.5 vs 52.8 ± 0.3, p<0.01). Diet quality was also positively (p<0.01) related to % energy from HCP foods. Saturated fat (p<0.01), added sugars (p<0.01), and sodium (p<0.02) intake were all inversely related to % energy from HCP foods while total fruit (p<0.01) and total vegetable (p<0.01) intake were positively related. BMI and risk of being overweight or obese were inversely related to the % of energy from HCP foods (both (p<0.01). The risk of high blood pressure was lower (p<0.01) as the % energy from HCP foods increased. These data demonstrate that a greater consumption (as % total energy) of foods that meet HCP is associated with better diet quality, BMI and blood pressure. Supported by the American Heart Association.
Lactoferrin modulates lipopolysaccharide‐induced perturbations in gene expression in human fetal intestinal epithelial cells
Authors: Nonnecke Eric B , Lönnerdal Bo L
Published: June 2021

Lactoferrin (Lf) functions in the orchestration of iron homeostasis, immune maturation, and proliferation of intestinal epithelial cells (IEC) in the breastfed infant. The exaggerated inflammatory response of enteropathogenic‐derived infantile diarrheas is potentiated, in part, by exposure to lipopolysaccharide (LPS). Lf sequesters the lipid A moiety of LPS with high affinity and may dampen the effects of overt TLR complex activation. In the present study, effects of Lf in the presence of LPS on the expression patterns of stress‐response genes were examined using a fetal IEC model. FHs74 cells were treated for 24 h with 400 μg/ml native (N)‐ and holo (H)‐ human (hLf), bovine (bLf), or commercially available bovine Lf (cbLf) with or without 1 μg/ml LPS. Cell viability was attenuated (> 30%) by LPS challenge, an effect that was ameliorated by the addition of Lf(s). Using PCR array profiling N‐hLf treatment was determined to differentially modulate LPS‐induced up‐regulation of numerous intermediates and products of the NFκB, MAPK, and Fas signaling pathways. Q‐PCR was employed to validate PCR array findings that Lf(s) mediates perturbations of multiple pro‐inflammatory cytokines due to LPS challenge including IL1, IL6, and IL8. Our findings suggest Lf may protect the neonatal intestinal epithelium against LPS by altering the expression of key regulatory genes associated with an LPS derived inflammatory response.

Grant Funding Source: N/A
Hepatic Stellate Cells Modulate the Phenotype of Liver Myeloid Dendritic Cells Rendering Them Tolerogenic
Authors: Dangi Anil , Sumpter Tina L , Matta Benjamin M , Stolz Donna B , Murase Noriko , Thomson Angus W , Gandhi Chandrashekhar R
Published: June 2021

Liver exhibits immune tolerance as illustrated by spontaneous acceptance of the allograft across MHC barriers in rodents and several cases of human transplantation. Since, rejections of the allografts still a major clinical problem, precise understanding of the mechanisms underlying the tolerogenic property of liver is important. Evidence indicates that the perisinusoidal hepatic stellate cells (HSCs) play an important role in hepatic tolerance by interacting with the immune‐regulatory cells. We hypothesized that HSCs might regulate migratory antigen‐presenting dendritic cells (DCs) in determining the fate of liver graft. We observed that myeloid (m) DCs co‐localize in vivo and in vitro with HSCs. Such association increases in transplantation due to ischemia/reperfusion‐induced disruption of endothelial barrier. In co‐culture, HSCs up‐regulated the expression of MHC II, CD80, CD86, and inhibitory ligand B7‐H1 on mDCs. The co‐culture also resulted in increased production of IL‐6 and IL‐10. HSCs suppressed the ability of mDCs to induce proliferation of allogeneic CD4+ and CD8+ T cells. HSC‐induced modulation of mDCs required cell‐cell contact, and was partially dependent on HSC‐derived MIP1á, and IL‐6‐induced STAT3 activation. The results suggest that HSCs play an important role in hepatic immunotolerance by altering the phenotype of mDCs, rendering them tolerogenic. (Supported by NIH PO1A1081678)
Role of PDGFRα in liver regeneration using hepatocytespecific knockout mice
Authors: Awuah Prince , Nejak-Bowen Kari , Monga Satdarshan
Published: June 2021

PDGFRα is classically known for its role in mesenchymal cells such as sinuoidal endothelial cells (SEC) of the liver. We have recently identified low expression of PDGFRα in adult hepatocytes, which is upregulated in hepatocellular cancer. To determine the role and regulation of PDGFRα in hepatocyte biology, we generated hepatocyte‐specific PDGFRα knockout mice (KO). Loss of PDGFRα in hepatocytes but not SEC was evident at 2 months, albeit no gross or histological anomaly was discernible. To further dissect the role of PDGFRα in hepatocyte proliferation and survival, we examined liver regeneration (LR) in KO and WT mice after partial‐hepatectomy (PH). A dramatic increase in total PDGFRα was evident at 24hrs after PH in WT, which was accompanied by tyrosine phosphorylation. KO mice maintained low PDGFRα levels at 24hrs although they did exhibit an increase over baseline suggesting ongoing signaling in SEC. Absent PDGFRα in hepatocytes did not lead to any changes in hepatocyte proliferation (PCNA) or viability (TUNEL) during LR at 24hrs or 48hrs after PH. Since molecular redundacy is well recognized during LR, we examined KO livers for any compensation after PH that may explain lack of a defect in LR process. A robust increase in phospho‐EGFR expression was observed in KO over the WT at 24hrs. Thus, while PDGFRα increase is clearly evident during LR, its loss in hepatocytes appears to be compensated by EGFR activation that may explain absence of overt defects proliferation or survival in KO. Funded by CATER fellowship (NIH T32 EB001026‐05) to PA; 1R01DK62277 and 1R01CA124414 to SPM
Tandem peptide blocks pemphigus vulgaris skin blistering in vivo and identifies a desmoglein receptor function leading to p38MAPK modulation
Authors: Spindler Volker , Kempf Bettina , Waschke Jens
Published: June 2021

In the life‐threatening autoimmune disease pemphigus vulgaris, epidermal blisters are caused by autoantibodies primarily targeting the cadherin‐type cell adhesion molecules desmoglein (Dsg) 3 and Dsg1 leading to loss of keratinocyte cohesion. Due to limited deeper insights in disease pathogenesis current therapy is restricted to relatively unspecific long term immunosuppression. Both, direct inhibition of Dsg trans‐interaction as well as altered intracellular signaling likely contribute to loss of cell adhesion. Here, we applied a tandem‐peptide (TP) in vivo consisting of two connected peptide sequences each targeting the Dsg adhesive interface. TP was capable to block autoantibody‐mediated direct interference of Dsg3 transinteraction as revealed by atomic force microscopy and optical trapping. Importantly, it was effective to completely abrogate autoantibody‐mediated skin blistering in mice. Relevant for a possible therapeutic use in patients, TP was effective also when applied topically. In addition, TP blunted autoantibody‐induced activation of p38MAPK, a central signaling pathway in pemphigus pathogenesis. These data indicate that p38MAPK links autoantibody‐induced direct inhibition of Dsg binding to skin blistering. By limiting both loss of Dsg trans‐interaction and subsequent p38MAPK activation, TP may serve as promising treatment option in pemphigus vulgaris. DFG SFB487
Recognition of Fungal β‐glucan by Human Neutrophil CR3 Results in Homotypic Aggregation and Neutrophil Extracellular Traps
Authors: Byrd Angel Shree’ , Yu Kebing , Johnson Courtney M. , Lavigne Liz M. , Salomon Arthur R. , Reichner Jonathan S.
Published: June 2021

The armament of neutrophil‐mediated host defense against fungal and bacterial pathogens includes the formation of a lattice of DNA and microbicidal enzymes known as Neutrophil Extracellular Traps (NETs). The receptor mediated interactions and intracellular signaling events responsible for the elaboration of NETs in response to fungi are not known and were determined in this study. Recognition of the fungal cell wall component β‐glucan by human neutrophils resulted in rapid formation of cell aggregates and NET release. This response required fibronectin + β‐glucan demonstrating a role for CD11b/CD18. CD11b/CD18 blockade obviated this response which did not require production of oxidative metabolites. Downstream signaling events were determined using a global phosphoproteomic approach. Upon fibronectin + β‐glucan treatment, 46 phosphopeptides were upregulated and 15 were downregulated as compared to fibronectin alone. Validation revealed a functional role for ERK (Y204). NET formation to Candida albicans hyphae was also shown to be dependent on CD11b/CD18 recognition of hyphal β‐glucan, occur in the absence of reactive oxidant species, and depend on ERK. This study successfully identified a key receptor ligand interaction that induces NET formation and elucidates the global tyrosine phosphoproteome of human neutrophils to gain a better understanding of innate immune responses to fungal pathogens.

NIH GM066194
Parents are not fully knowledgeable of their children's experiences of food‐insecurity
Authors: Escobar-Alegría Jessica L , Frongillo Edward A , Fram Maryah S , Pérez-Garay Melly , Macauda Mark M , Billings Deborah L
Published: June 2021

Although children and adults experience food insecurity differently, it is commonly assumed that parents are knowledgeable about their children's experiences. To examine this assumption, we explored and assessed parent knowledge of child food insecurity experiences in 16 Hispanic food‐insecure families. Separate in‐depth interviews were conducted in Spanish with a child, the mother, and another adult (when applicable). Interview data were analyzed thematically, applying previous research showing that children experience food insecurity through domains of awareness and responsibility. We compared child versus adult reports of what children experienced. Parents were fully knowledgeable of children's cognitive and emotional awareness of food insecurity in only 7 out of 16 families. Although half of children had initiated actions to help manage food insecurity, no parents were aware of this. No parents were knowledgeable of children's physical hunger or efforts to generate new food resources. Overall, parents had incomplete or inaccurate information of children's experiences of food insecurity; this is of serious concern since parental report is the basis for current monitoring and estimation of the prevalence of child food insecurity and hunger in the U.S. Funded by University of South Carolina.
Provitamin A Absorption and Conversion from a Unique High Beta‐Carotene Tomato is Higher when Consumed with Avocado
Authors: Kopec Rachel E , Cooperstone Jessica L , Schweiggert Ralf M , Riedl Kenneth M , Harrison Earl H , Francis David M , Clinton Steven K , Schwartz Steven J
Published: June 2021

It has been widely demonstrated that dietary lipid increases carotenoid absorption in humans. In contrast, surprisingly little is known about the effect of lipid on the conversion of provitamin A carotenoids from a typical vegetable meal. We hypothesized that the consumption of provitamin A (from a novel high β‐carotene tomato variety or carrots) with lipid rich avocados would promote the absorption of provitamin A carotenoids and deliver greater quantities of vitamin A as compared to the same meal without avocado. We performed two separate human clinical studies (n=12) with a randomized, crossover design where we quantitated β‐carotene, α‐carotene, lutein, and multiple retinyl esters in the immediate post‐prandial triglyceride rich lipoprotein fraction (TRL) fraction of blood plasma. After a two week washout, the test food was consumed with or without avocado, and blood samples were taken over 12 hours. TRL fractions were immediately isolated, extracted, and quantitated by High Performance Liquid Chromatography‐Mass Spectrometry. A greater level of carotenoid was absorbed when the test meals were consumed with avocado as compared to no avocado. We expect that provitamin A carotenoid conversion to retinyl esters will also be enhanced by co‐consumption of dietary lipid. These findings provide new information on the effects of fat on carotenoid absorption and metabolism in humans to deliver vitamin A.
Challenges to making local food accessible through farmersˈ markets and community supported agriculture: DeKalb County, Georgia
Authors: McAuliffe Corey , Whitaker Laura , Girard Aimee Webb
Published: June 2021

Local farmers and producers face many challenges when trying to harvest and distribute their goods. Farmers’ markets and community supported agriculture (CSA) offer an alternative distribution system that has yet to be systematically assessed within metro Atlanta.

We partnered with Georgia Organics (a non‐profit dedicated to ensuring healthy, sustainable, and locally grown food for all Georgians) in order to assess issues faced by local producers in product accessibility. We completed interviews with farmers’ market managers, farmers, CSA managers, and a cooperative market manager to assess the challenges and successes of product accessibility through famers’ markets and CSAs within DeKalb County, Georgia. Research was completed to inform DeKalb County Board of Health initiatives. Qualitative data was collected on system operations, accessibility to local foods, challenges and barriers to the system, motivating factors, and sustainable system components.

Data shows financial barriers to production, prohibitive policies, limitations for marketing and distribution systems, and limiting environmental factors. In order to increase access to local, healthy foods in low income areas of DeKalb County, local and state level government can reduce producers’ risk through funding and supporting adapted models. This study was supported by the Emory University Office of University‐Community Partnerships.

Grant Funding Source: Emory University Office of University‐Community Partnerships
Food Availability is Related to Body Mass Index of Children in an Urban Area of Brazil
Authors: Martins Paula Andrea , de Carvalho Cremm Elena , de Oliveira Maria Aparecida , Gittelsohn Joel
Published: June 2021

Introduction

There is little research about the role of the neighbourhood food environment in the obesity epidemic in developing countries. We assessed the effects of food availability on nutritional status of the children.

Methods

Nutritional status of children under 10 years (n=531) was assessed in a household survey in 35 census tracts of the urban area of the city of Santos, Brazil (2010). Food availability of all stores (n=672) located in each census tract was assessed using a validated instrument. A Healthy Eating Score was calculated for each store according to the degree of industrial processing and the nutrient content of available food items. Stores were classified as More or Less Healthy Eating Promoter (MHEP and LHEP) using the median as a cutoff point. Food stores and household variables were aggregated to the census tract level.

Results

In Multivariate Linear Regression models, Children's BMI for age was inversely associated with the proportion of LHEP stores (β= −0.53, P=0.002) adjusting for household SES (R2=0.25, p=0.008).

Conclusion

Policies to improve food environment in developing countries are needed to address the growing problem of obesity in children.
Using Technology to Assess gain in Nutrition Knowledge Among the Youth
Authors: Roy Heli
Published: June 2021

The extension service in Louisiana offers nutrition education programs in schools across the state. We tested for change in nutrition knowledge among elementary school students based on 9 nutrition lessons delivered on various topics in nutrition during the school year using Turning Technologies audience response system. The enrichment classes varied from 20 minutes to 40 minutes in length. The students received informational fact sheets after the lessons and food tasting was conducted 50% of the time. We developed 10 pre and post test questions based on the lesson delivered to test students gain in nutrition knowledge. Pre and post test questions were delivered using Turning Point Student Response System software and the students used clickers to deliver the answers and the data was collected directly by the receiver. There were a total of 226 pre and posttests completed in six elementary schools. The students were enthusiastic about pre and post assessment and accepted the devices immediately. The results show that there was a highly significant gain in nutrition knowledge among the youth participating in the nutrition education programs (P<.0001) overall.
Modulation of Shh signaling by microRNAs
Authors: Xu Yanhua , Hu Diane , Barbeau Rebecca , Barczak Andrea , Marcucio Ralph
Published: June 2021

Sonic hedgehog (Shh) exhibits a dynamic expression sequence in the head. First, Shh is detected in the forebrain and then Shh is induced in the facial ectoderm where it forms a signaling center, the Frontonasal Ectodermal Zone (FEZ) that regulates morphogenesis of the upper jaw. Induction of Shh in the FEZ is regulated by SHH from the brain, but the mechanisms underlying this induction pattern is unknown. Here, we examine the extent to which the SHH pathway is regulated by miRNAs. Microarray analysis revealed that miR199b was expressed at lower levels in the FEZ at HH18 compared to HH22, and was up‐regulated in response to exogenous SHH. In situ hybridization at HH18 revealed that miR199b was expressed broadly in the future Shh domain of the FEZ, and by HH21 became restricted to the edge of the Shh domain in the FEZ. A similar complementary expression pattern between Shh and miR199b was found in the floorplate of the neural tube. Over‐expressing mir199b in embryos caused narrowing of the face that was associated with decreased Shh expression. There was no obvious increase in apoptosis, but we observed reduced cell proliferation in the mesenchyme. To assess how miR199b affects SHH signaling, we determined that infection of RCAS::mir199b in DF‐1 cells inhibited SHH signal activation and cell proliferation. Taken together, these results suggest that miR199b acts a negative regulator of SHH signaling in embryos and cultured cells.

Grant Funding Source : This work was funded by grant from the National Institutes of Health (5R01‐DE018234 to R.M.).

MicroRNA array was supported from the Sandler Asthma Basic Research (SABRE) Center Functional Genomics Core Facility.
TWISTing a Hand on the mandible
Authors: Ruest L. Bruno , Zhang Yanping , Blackwell Evan L. , McKnight Mitchell T. , Knutsen Gregory R.
Published: June 2021

Formation of the mandible requires the exquisite control of neural crest cell (NCC) development. Transcription factors controlling the early proximo‐distal development of the mandibular NCC are mostly elusive. We hypothesized that the interacting basic helix‐loop‐helix (bHLH) transcription factors are involved. One of these factors is the Saethre‐Chotzen gene Twist1. To investigate the function of Twist1 during mandibular development, we conditionally inactivated the gene at E9.5 in the mandibular arch NCC using the Hand2‐Cre mouse line. The mandible in the Twist1 mutant embryos was shorter due to the reduction of the proximal articular elements. The expression of several markers of lower jaw development appeared to be normal, including the bHLH factors Hand1 and Hand2. Unexpectedly, mandibular mineralization was affected in the heterozygous embryos, likely due to an imbalance between Twist1 and Hand2 to regulate Runx2 function. The expression of miR‐199 and 214 was normal in the mutant embryos and our results revealed that Hand2 is also controlling the expression of these microRNAs. These results suggest that Twist1 and Hand2 interact during mandibular development, but each factor may also have independent functions that depend on its own interactions with other factors or themselves. Further studies are needed to understand the role of the interaction between Twist1 and Hand2 in mandibular development.
Artificial neural network scaffold transplanted promotes the recovery of structure and function after rat spinal cord transection
Authors: Zeng Yuan-shan
Published: June 2021

Functional deficits after spinal cord injury attribute to neural cell death and loss of connectivity. Here we designed tissue engineered neural networks acting as bridges to rebuilt signal connection between the cut ends of the ruptured spinal cord. Mesenchymal stem cells (MSCs) with genetically enhanced expression of TrkC were induced into cholinergic neurons after cocultured with Schwann cells overexpressing neurotrophin‐3 in a gelatin sponge scaffold for 14 days. The formation of neural networks was confirmed by electron microscope and recording of spontaneous postsynaptic currents by whole‐cell patch clamp. Then, these neural network scaffolds were grafted acutely into rats with removal of 2 mm spinal cord tissue. Eight weeks later, MSCs‐derived neurons of the grafts maintained their synaptic connection or formed new connection with regenerative axons. Although Biotinylated Dextran Amine labeled fibers of corticospinal tract only formed a few connections with MSCs‐derived neurons in the grafts, 5‐HT labeled fibers formed much more connections. Furthermore, a part of these connections were confirmed to be synaptic connections by double‐labeled immunoelectron microscope. Animals with neural networks scaffold transplantation resulted in higher BBB score and improved cortical motor evoked potential. These findings indicated that MSCs‐derived neural networks integrated into host neural networks
Retinoic Acid Antagonism of Fgf8 during Forelimb Development
Authors: Cunningham Thomas James , Duester Gregg
Published: June 2021

Retinoic acid (RA) in the proximal limb is purported to act as a proximal signal during proximodistal patterning of chick limbs, however this remains controversial as it is not supported by genetic findings in mouse. Using a RARE‐lacZ RA‐reporter transgene (which we show to be sensitive to 2.5 nM RA) we found that mouse embryos lacking retinol dehydrogenase‐10 (Rdh10) lost all RA activity in fore‐ and hindlimb fields although they retain RA activity in posterior neuroectoderm. Although forelimbs were stunted, Rdh10 mutant hindlimbs were normal, and both expressed key patterning genes in the correct spatiotemporal pattern. Previous studies on Raldh2 mutants have shown that RA antagonizes Fgf8 and that excessive FGF signaling in both primitive streak and cardiac domains blocks forelimb initiation. Here, we show that Rdh10 mutants exhibit ectopic Fgf8 expression only posterior to the heart where the forelimb field arises, suggesting a mechanism for how stunted forelimbs arise. We propose that while RA does not pattern the forelimb itself, axial heart RA‐Fgf8 antagonism is required prior to forelimb field formation to provide an environment permissive for normal budding and patterning of the forelimb.
Soft tissue calcification in the Ossabaw miniature pig: experimental and kinetic modeling studies
Authors: Wastney Meryl , Lee WangHee , Jackson George , Alloosh Mouhamad , Sturek Michael , Lachcik Pam , Martin Berdine , Weaver Connie M
Published: June 2021

Early stage deposition of calcium into soft tissues is a pre‐cursor to several chronic diseases and an appropriate large animal model is needed. The Ossabaw pig develops metabolic syndrome when fed excess calorie atherogenic diet and progresses to type 2 diabetes and atherosclerosis. Male pigs were fed for 5 months with chow diet (health, lean; n=7) or atherogenic diet (n=8) consisting of chow supplemented with 2% cholesterol, 43% kcal from fat, and 20% kcal from fructose. Pigs were verified to have metabolic syndrome (MetS) by obesity, insulin resistance, impaired glucose tolerance, dyslipidemia, and hypertension. All pigs received intravenous injection of 50nCi/kg of 41Ca and allowed to recover for 1 month, then sacrificed. Blood was drawn at time 0, nearly hourly for the first 24 h, and 1, 2, 3, 6, 8, 10, 15, 20, and 28 days after injection. Peripheral artery segments were harvested from each pig at 4 time points over the 28d recovery. At sacrifice the MetS swine had detectable 41Ca in coronary arteries, but the lean controls had extremely low levels of 41Ca. Intravascular ultrasound showed calcification in MetS, but not lean pigs. A compartmental model of calcium metabolism was developed and deposition into arteries estimated. Through tissue acquisition and kinetics, this pig provides a humanoid model for studying vascular calcification.
Palmitoleate Supplementation Dissociates Liver Inflammatory Response from Hepatic Steatosis in Mice
Authors: Guo Xin , Li Honggui , Xu Hang , Meng Cong , Wu Chaodong
Published: June 2021

Supplementation of monounsaturated fatty acids generates beneficial effects. We investigated the effects of palmitoleate supplementation on liver inflammatory and metabolic responses. At 5 – 6 weeks of age, male wild‐type C57BL6/J mice were fed a low‐fat diet (LFD) for 12 weeks and supplemented with palmitoleate (600mg/kg/d), oleate (600mg/kg/d), or bovine serum albumin (BSA) via oral gavages for the last 4 weeks. Compared with controls, palmitoleate supplementation increased circulating levels of palmitoleate and improved systemic insulin sensitivity. Locally, palmitoleate supplementation increased hepatic fat deposition and the expression of sterol regulatory element‐binding protein 1c and fatty acid synthase. However, palmitoleate supplementation reduced the numbers of macrophages/Kupffer cells in livers of the treated mice. Consistently, supplementation of palmitoleate, but not oleate or BSA, decreased the phosphorylation of nuclear factor kappa B and the expression of tumor necrosis factor alpha and interleukin‐6 in livers. Together, these findings demonstrate a novel role for palmitoleate supplementation in dissociating liver inflammatory response from hepatic steatosis. When used with anti‐steatosis agents, palmitoleate supplementation may be a viable approach to preventing liver inflammation.

Grant Funding Source: ADA