bluuebunny/crossref_metadata_2025 huggingface dataset:
Identifying the Expression Patterns of xCT in Zebrafish to Determine its Role in Neuroregeneration
Authors: Solorzano Marissa A , Ladd Nicole A , Franz Kevin C , Da Silva Christopher O , Degnan Shannon M , Chase Leah A , Putzke Aaron P , Krueger Brent P
Published: June 2021

System xc− is a heterodimeric amino acid transporter comprised of a transmembrane light chain unit, xCT, and an extracellular heavy chain unit, 4F2HC. System xc− has been shown to exchange intracellular glutamate for extracellular cystine, which is then reduced within the cell to cysteine, the limiting reagent for glutathione production. Glutathione is a reducing agent that is important in reducing oxidative stress, which untreated can trigger cell death. It has been shown that system xc− is strongly expressed in the central nervous system, particularly in neuroprotective cells such as astrocytes and microglia. It is believed the reduction of oxidative stress in the environment of neurons and neuroprotective cells is critical in processes such as neuroregeneration. The current focus of this study is to qualitatively determine the expression patterns of the xCT gene in zebrafish embryos using in situ hybridization; preliminary results will be presented. This is in preparation for exploring the role that xCT plays in neuroregeneration using laser ablation to mimic spinal cord injury in vivo.

Support or Funding Information

National Science Foundation award 1058981.
Activation of Akt increases Cell Surface Expression of System xc−
Authors: Versluis Philip , Gibson Amanda , Schmidt Mackenzie , Smith Daniel , Chase Leah
Published: June 2021

System xc− is a heterodimeric plasma membrane transporter involved in the exchange of intracellular glutamate for extracellular cystine. As such, this transporter plays a critical role in the production of the antioxidant glutathione. Previous studies in our lab have demonstrated that there is an increase in cell surface expression within ten minutes of exposure to H2O2 in confluent U138MG human glioma cells. The study described herein sought to begin to characterize the mechanism by which H2O2 regulates the trafficking of xCT. We hypothesized that Akt signaling is necessary for H2O2‐mediated trafficking of of xCT. A significant increase in Akt phosphorylation was observed in U138MG cells following ten‐minute exposure to 3 mM H2O2 compared to vehicle‐treated cells using western blot analysis. Treatment with the Akt inhibitor 10‐DEBC (2.5μM) for 30 minutes prior to and during H2O2 exposure resulted in a decrease in H2O2‐induced phosphorylation of Akt at Ser473. Similar inhibition of Akt phosphorylation at Thr308 was observed following treatment of cells with 1.0μM API‐2. Next, we used simultaneous treatment of cultured glioma cells with both inhibitors in the presence of H2O2 to determine if such treatment led to a reduction in the trafficking of endogenously expressed xCT to the plasma membrane. Our preliminary data suggests that Akt activation is necessary for H2O2‐induced trafficking xCT to the plasma membrane in cultured glioma cells. To determine if the regulation of xCT cell surface expression is not limited to human glioma cells where xCT is oven overexpressed, we also studied the role Akt plays in the trafficking of recombinant, transiently‐expressed xCT in COS‐7 cells. COS‐7 cells transfected with myc‐tagged xCT, 4F2HC and a constitutively active form of Akt showed higher levels of xCT localized to the membrane compared with cells transfected with a dominant negative form of Akt. Collectively, these data suggest that Akt is an important regulator of xCT cell surface expression.

Support or Funding Information

This work was supported by NSF‐RUI 0843564.
Relationship between Solid Food Introduction and Picky Eating in the STRONG Kids 2 Cohort
Authors: Cole Natasha Chong , Lee Soo‐Yeun , Donovan Sharon M.
Published: June 2021

Picky eating is prevalent in toddlers and has been linked to infant feeding practices. Early introduction of solid food is associated with picky eating; however, the relationship with specific types of complementary food is unknown. The objective of this study was to examine associations between types of solid food introduced during infancy and picky eating behavior in children 18‐months of age. Participants were drawn from an ongoing STRONG Kids 2 birth cohort of mother‐infant dyads (current study n = 259, full sample n = 440). Mothers responded to survey questions about the type and amount of solid food introduced during infancy, the perception of their child's picky eating behaviors at 18‐months, and whether they breastfed or formula‐fed their child during the introduction to solids. Ordinal logistic regression (i.e., proportional odds model) was used to determine associations between types of solid food introduced and odds of picky eating, while controlling for child sex, race/ethnicity, age of introduction to solids, and mode of milk feeding. The introduction of sweets (e.g., ice cream and pudding), cereal, pureed fruit, and egg yolk were positively associated with parental perception of picky eating at 18‐months (all p<0.05). There was no association between parental perception of picky eating and the introduction of vegetables, meat, or dairy. Overall, our findings suggest that early exposure to various tastes and textures may influence the development of picky eating later in childhood. Further research is needed to determine the influence of the timing and type of solid food introduction on children's long‐term dietary variety and eating behavior.

Support or Funding Information

Supported by National Institute of Food and Agriculture, U.S. Department of Agriculture, under award number 2011‐67001‐30101 and the Dairy Research Institute.
Culturally‐Relevant Diabetes Education Program Viewed Favorably by Rural‐Residing Latinos
Authors: Keane Michelle , Bahl Morgan , Francis Sarah L
Published: June 2021

Rural‐residing Latinos are at high risk for diabetes and diabetes complications. Culturally‐relevant diabetes education is needed for this group. Latinos Living Well (LLW) was created using the Social Marketing Theory (SMT) to ensure it was audience‐centered and culturally‐sensitive for rural‐residing Latinos in a Midwest state. LLW is a four‐week diabetes education program focused on diet and exercise. The two‐hour classes include culturally‐relevant physical activity and cooking practicums, group activities and goal setting. Classes were taught in Spanish and delivered through Cooperative Extension. This study evaluated participant satisfaction and perceptions of LLW. A convenience sample of rural‐residing Latino adults ages 18+ years (n=120) were recruited through word of mouth, flyers, and radio ads; 76 completed the program (63.3% completion rate). Participants completed sociodemographic questions at Class 1 and a qualitative program evaluation at the end of Class 4. The majority of participants were first generation immigrants (n=63; 87.5%), females (n=73; 97.3%), ages 30–49 (n=34; 51.6%), with at least 9–12 years of education (n=56; 77.8%). A diabetes or pre‐diabetes diagnosis was not required for enrollment into LLW; however, two‐thirds (n=49; 65.3%) reported a history of diabetes. The evaluation asked about participants' perceptions of LLW and what lifestyle changes were made as a result of attending. Descriptive statistics analyzed sociodemographic and evaluation data. Nearly all (n=69, 94.5%) believed LLW helped them to make healthy lifestyle changes. These changes included preparing at least one class recipe at home (n=57, 79.2%) and being more physically active (n=58, 81.7%). The content was viewed as understandable (n=72, 98.6%). The majority (n=70, 95.8%) rated LLW as “excellent” or “good” and would recommend the program to a friend (n=72, 94.7%). Results suggest rural‐residing Latinos enjoyed LLW, thus indicating using culturally‐relevant examples and activities helped participants view the program favorably. Furthermore, it is likely participants will refer friends and family, be motivated to apply the information they learned and attend other health programming through Cooperative Extension.

Support or Funding Information

This research was funded by the United States Department of Agriculture, National Institute of Food and Agriculture Rural Health and Safety Education Grant, #2014‐46100‐22349
Development and Validation of a B.E.A.N. Survey for College Students
Authors: Sowers Morgan , Colby Sarah , Allison Chelsea , Zhou Wenjun
Published: June 2021

Background

College students often do not consume the recommended amounts of beans and legumes. To develop interventions aimed at improving the intake of beans and legumes in the college population, more information is needed on the behaviors, environment, attitudes, and nutrition knowledge (B.E.A.N.) of college students regarding beans. Validated tools to assess these bean‐related constructs are currently lacking.

Objectives

The objectives of this study are to 1). Gather preliminary qualitative information about bean‐related constructs (Phase I); and 2). Create a valid and reliable survey tool to measure the B.E.A.N. constructs with a larger sample of college students (Phase II).

Methods

Phase I included cognitive interviews with university students (n = 30) to collect baseline data on the B.E.A.N. constructs. Phase II involved survey development and content validation with 5 content experts and face validation with 5 undergraduate students. A convenience sample of undergraduate students was surveyed to test internal structure and consistency (n = 355), establish convergent and discriminant evidence (n = 351), and measure test‐retest reliability (n = 242) of the developed tool.

Results

Development and revisions in Phase I and content and face validation resulted in a 13‐question survey consisting of four primary constructs of behavior (4 items), environment (2 items), attitudes (5 items), and nutrition knowledge (2 items). In Phase II, factor analysis resulted in one factor of “bean consumption” being created from combining attitude and behavior questions, with factor loadings from .776 to .947. However, two environment and two knowledge questions were retained due to perceived relevancy. The internal consistency was α = 0.967 for the “bean consumption” construct and α = 0.962 for the entire survey. The knowledge construct was validated by those that had taken an introductory nutrition course having significantly higher mean knowledge scores than those that had not taken an introductory nutrition course (p < 0.001; 3.05 ± .89 vs. 2.30 ± 1.62, respectively). The test‐retest results ranged from r = .480 to .825 with all correlations being statistically significant (p < 0.010; 2‐tailed). The average total score for the pilot testing was 34.74 (± 27.27) out of a possible 100 points.

Conclusion

This validated survey may be used in future research to examine the B.E.A.N. of college students regarding beans. This knowledge may be valuable in developing future effective health promotion interventions with college students.

Support or Funding Information

This project was supported by Agriculture and Food Research Initiative Grant no. 2014‐67001‐21851 from the USDA National Institute of Food and Agriculture, “Get Fruved:” A peer‐led, train‐the‐trainer social marketing intervention to increase fruit and vegetable intake and prevent childhood obesity – A2101.
Impact of long‐term dietary patterns and short‐term nutrient intake on the gut microbiota of children 4 to 8 years of age
Authors: Berding Kirsten , Donovan Sharon
Published: June 2021

The gut microbiota is undergoing rapid changes in the first few years of life and diet plays a fundamental role in shaping the gut microbiota. It is commonly believed that by age 3 years, the microbiota composition resembles that of an adult. However, recent research shows differences in the microbiota composition between pre‐adolescence children and adults. Little is known about how diet influences the gut microbiota composition in young, healthy children. Herein, the effect of long‐term dietary patterns and short‐term dietary intake on longitudinal changes in microbiota composition were studied in children aged 4–8 years. A food frequency questionnaire, 3‐day food record and stool sample were collected for each participant (n=24) at baseline, 6‐weeks and 6‐months post‐baseline. Nutrient intake was analyzed using Nutrition Data System for Research (NDSR). Dietary patterns based on food groups were identified using principal component analysis. Bacterial genomic DNA was analyzed for total bacteria, Prevotella, Lactobacillus, and Roseburia by quantitative real‐time qPCR. Distinct microbial profiles were identified for three long‐term dietary patterns. Compared to the other two dietary patterns, a diet pattern characterized by a higher intake of fruits, vegetables, meat, fish, legumes, nuts and seeds, but also sugary drinks and desserts showed higher abundance of total bacteria. A diet pattern defined by a higher intake of whole grains and starchy vegetables was characterized by lower density of Lactobacillus and Roseburia, but higher density of Prevotella. Lastly, a diet pattern characterized by higher intake of refined grains, French fries and a lower intake in salty snacks was associated with lower abundance of total bacteria. Short‐term dietary fiber intake was not significantly correlated with bacterial abundance. Overall, there was no significant difference in the mean of the bacteria concentration between baseline and 6 weeks. However, bacterial abundance differed for some individuals between the two time points. In conclusion, long‐term dietary patterns influence the fecal microbiota of children aged 4 to 8 years. Analysis of 6 month samples will inform findings relative to the longitudinal stability of the gut microbiota. Future analysis includes microbiome analyses using next generation sequencing and assessment of short chain fatty acid concentrations.

Support or Funding Information

USDA Hatch Grant ILLU‐698‐311
Vitamin K, Vitamin D, and Lower Extremity Function: Results from the Osteoarthritis Initiative and Health, Aging and Body Composition Studies
Authors: Shea Kyla , Loeser Richard F , Kritchevsky Stephen B , Houston Denise K , McAlindon Timothy E , Booth Sarah L
Published: June 2021

Knee osteoarthritis (OA) is the leading cause of impaired lower‐extremity function in older adults. Vitamin K insufficiency has been associated with greater structural knee OA progression, but studies associating vitamin K status with functional consequences of knee OA are lacking. The purpose of this study was to determine the association between vitamin K intake and lower‐extremity function in the Osteoarthritis Initiative (OAI), a prospective observational study of knee OA (n=4497, 58% female, 61±9 years old). Because the mechanisms underlying the role of vitamin K in joint health also implicate vitamin D, we additionally evaluated the association of combined vitamin K and vitamin D intake with lower extremity function. Dietary intakes were estimated using a food frequency questionnaire at baseline. Lower extremity function was assessed at baseline and annually over 4 years using the 20‐meter gait speed and time to complete 5 chair stands. Participants in the highest quartile of vitamin K intake had significantly faster 20‐meter gait speed and chair stand time over 4 years compared those in the lower quartiles (both p‐trend<0.001; analyzed using linear mixed models adjusted for demographics, co‐morbidities, diet and lifestyle characteristics). When vitamin K and D intakes were considered together, those in the highest quartiles of both vitamin K (≥229 mcg/d) and vitamin D (≥600 IU/d) intake had significantly faster gait speed (p<0.001) and chair stand time (p=0.002) over 4 years compared to participants in the highest quartile of either nutrient alone and compared to participants in the lowest quartile of both nutrients. We sought to replicate these findings, given their novelty, in the Healthy, Aging and Body Composition Knee OA sub‐study (n=1102, 60% female, 75±3 years old). Vitamin K and D status were assessed using circulating vitamin K and 25(OH)D at baseline and lower extremity function was assessed using the short physical performance battery (SPPB) and 20‐meter gait speed at baseline and over 4–5 years of follow‐up. Participants with both sufficient vitamin K (≥1.0 nM) and 25(OH)D (≥50nM) had significantly better SPPB scores and faster gait speed over follow‐up compared to those with sufficient levels of either nutrient alone and compared to those who were insufficient in both (p=0.001 and 0.005 respectively; adjusted for demographics, season, co‐morbidities, diet and lifestyle characteristics). These findings suggest vitamin K and vitamin D may be mutually beneficial to lower‐extremity function related to knee OA but need to be confirmed in clinical trials.

Support or Funding Information

Supported by the USDA (Cooperative Agreement 58‐1950‐7‐707) and the NIH (grants K01AR063167, P30AG21332, R01AG028050, R01NR012459, contracts N01AG62101, N01AG62103, N01AG62106)
Calcium intake below the recommended dietary allowance is associated with lower tibia bone mineral content and strength in young adults entering initial military training
Authors: Nakayama Anna , Lutz Laura J , McClung James P , Gaffney‐Stomberg Erin
Published: June 2021

Background

Calcium (Ca), a critical micronutrient for bone health, has been deemed a nutrient of public health concern due to consumption at levels below the recommended dietary allowance (RDA). Stress fracture risk is elevated during initial military training (IMT) and supplemental Ca during IMT reduces stress fracture incidence.

Objective

To assess dietary Ca in young adults entering IMT and to determine whether habitual Ca intake is associated with indices of bone health at the tibia, a commonly fractured site.

Methods

A retrospective analysis was performed using baseline data from 3 randomized control trials that assessed the efficacy of Ca and vitamin D supplementation for optimizing bone health during IMT in Army, Air Force, and Marine recruits. Ca intake was estimated from a semi‐quantitative food frequency questionnaire and adequacy was based on the age‐specific RDAs. Bone health was evaluated by peripheral quantitative computed tomography (pQCT) of the tibia at the metaphysis and diaphysis. The associations between pQCT parameters and Ca intake adequacy were assessed by multivariate regression (SPSS, v21) controlling for age, sex, race, self‐reported physical activity, BMI, smoking, education level, and use of Depo Provera.

Results

Of the 399 study participants (n=223 females, n=180 males), 251 (62%) did not meet the RDA for Ca. Adequate Ca intake was positively associated with total bone mineral content (BMC; B=.091, p=.005) and strength (B=.084, p=.020) at the metaphysis. When participants were stratified by sex, 70% of females and 52% of the males did not meet the RDA. In the female cohort, adequate Ca intake was positively associated with BMC (B=.169, p=.011) and strength (B=.142, p=.033) at the metaphysis and cortical BMC (B=.134, p=.042) at the diaphysis. There were no associations in the male cohort.

Conclusion

Adequate Ca intake in young adults is positively associated with measures of tibia BMC and strength. As recruits with Ca intake below the RDA at the start of training may experience diminished bone health potentially leading to greater injury susceptibility, adequate Ca intake should be emphasized prior to initiating military training.

Disclaimer: The views expressed in this abstract are those of the authors and do not reflect the official policy of the Department of Army, Department of Defense, or the U.S. Government.

Support or Funding Information

Research supported by USAMRMC, DHP, and in part by an appointment to the Postgraduate Research Participation Program administered by the Oak Ridge Institute for Science and Education.
High Oleic Soybean Oil Improves Cardiometabolic Health in Adults
Authors: Baer David J , Henderson Theresa , Gebauer Sarah K
Published: June 2021

High oleic soybean oil (HOSBO) is a shelf‐stable oil that provides broad versatility for both solid fat and liquid oil applications for food preparation. The impact of replacement of HOSBO for alternative solid fat and liquid oils on cardiometabolic risk factors is largely unknown. This randomized, diet‐controlled, crossover trial was conducted with four treatments: 1) HOSBO, 2) soybean oil (SBO), 3) 80:20 blend of HOSBO and fully hydrogenated SBO (FHSBO), and 4) 50:50 blend of palm oil and palm olein (“palm blend”) to measure the lipid, lipoprotein, and other cardiometabolic factor responses to use of these oils under typical food preparation methods. Subjects (n=60) were randomized to a sequence of the 4 treatments and fed a controlled diet containing 50% of their total fat from the treatment (total dietary fat = 35% of energy). At the end of each 29‐day treatment period, blood samples were collected on 2 days and a 24 hr urine sample was collected. Concentrations of atherogenic markers [LDL cholesterol (LDLc), ApoB, Non HDL cholesterol, LDL particles] were higher after consumption of the palm blend compared to HOSBO and the other treatments (overall treatment P <0.0001). LDLc and ApoB were higher after consumption of the HOSBO compared to SBO (P=0.01, P=0.02, respectively). There was no significant difference in these atherogenic markers between the HOSBO+FHSBO blend and the HOSBO. Thus, replacement of up to 20% of HOSBO with FHSBO provides solid fat functionality with no change in atherogenic markers. Consumption of the palm blend increased HDL cholesterol (HDLc) and ApoAI compared to all other treatments, with no differences among the other treatments for HDLc and ApoAI. Ratios of total cholesterol‐to‐LDLc and LDLc‐to‐HDLc were highest after consumption of the palm blend compared to the other treatments (P <0.0001). Triglycerides, fasting glucose, and blood pressure were similar across all 4 treatments. HOSBO, an important option for replacement of solid fat and liquid oils in multiple food applications, maintains or improves lipid and lipoprotein profiles in humans compared to alternative functional fats.

Support or Funding Information

Funded by USDA, ARS and Qualisoy
Comparison of the Impact of High‐Dose DHA and EPA Supplementation on the Intravascular Kinetics of Very‐Low Density Lipoprotein Apolipoprotein C‐III
Authors: Vors Cecile , Johanne Marin , Allaire Janie , Tchernof André , Couture Patrick , Lamarche Benoit
Published: June 2021

Background

Apolipoprotein C‐III (apoC‐III) is an emerging risk factor for cardiovascular disease (CVD) and its presence on very‐low density lipoproteins (VLDL) may enhance their atherogenicity, primarily by delaying their rate of catabolism. We have shown in a recent study that high‐dose supplementation with docosahexaenoic acid (DHA) led to more pronounced reductions in TG compared with eicosapentaenoic acid (EPA), but mechanisms remain unclear. The objective of this study was to compare the individual effect of EPA and DHA on VLDL‐apoC‐III metabolism in men and women at risk of CVD.

Methods

In a randomized double‐blind crossover trial, 10 men and 8 women with abdominal obesity and low‐grade inflammation were subjected to three 10‐wk supplementation phases: 1) EPA (2.7 g/d); 2) DHA (2.7 g/d); 3) corn oil (0g DHA, 0g EPA, control), separated by a 9‐wk washout. All supplements were provided as 3×1g capsules for a total of 3g/d. After each dietary intervention, subjects received a primed‐constant infusion of [5,5,5‐D(3)]‐L‐leucine for 12h under constant feeding conditions for the determination of VLDL‐apoC‐III kinetics.

Results

Both DHA (−11.7%, p=0.01) and EPA (−6.4%, p=0.03) similarly reduced the fractional catabolism rate (FCR) of VLDL‐apoC‐III compared with control (p=0.75 between DHA and EPA). On the other hand, neither DHA nor EPA supplementation had a significant impact on VLDL‐apoC‐III concentrations and production rate (PR). Compared with control, DHA‐ and EPA‐induced reductions in TG were correlated with DHA‐ and EPA‐induced variations in VLDL‐apoC‐III PR (r=0.49, p=0.03 and r=0.48, p=0.05 respectively), but not with changes in VLDL‐apoC‐III FCR.

Conclusions

High‐dose supplementation with DHA or EPA is associated with reduced clearance of VLDL‐apoC‐III, with no effect, however, on VLDL‐apoC‐III concentrations and production. Although the DHA‐ and EPA‐induced reductions in serum TG may be more sensitive to treatment‐induced variations in VLDL‐apoC‐III production, other mechanisms must be investigated to explain why DHA may reduce serum TG more than EPA.

Support or Funding Information

Financial support for this RCT was provided exclusively by a grant from the Canadian Institutes for Health Research (MOP‐123494). Douglas Laboratories provided the EPA, DHA and placebo capsules used in this study. The funding source was not involved in the study design, conduct of the study, or collection, management, analysis, or interpretation of the data or in the preparation or review of the manuscript and had no right to approve or disapprove of the submitted manuscript. CV is a fellow of the French Foundation for Medical Research (FRM, file code: 40303). JA is a recipient of a PhD Scholarships from the Canadian Institutes for Health Research and the Fonds de recherche du Québec – Santé.
Effects of Dietary Resveratrol on Gut Microbiota in Mice with Colitis
Authors: Li Fang , Gu Min , Cai Xiaokun , Song Mingyue , Li Zhengze , Goulette Tim , Xiao Hang
Published: June 2021

Accumulating evidence indicated that gut microbiota play important roles in the pathogenesis of colitis, and microbiota composition could be modulated by dietary components. Therefore, ameliorating colitis‐associated bacterial dysbiosis by dietary components may be a unique strategy to improve gut health. Herein, we determined the effects of resveratrol on gut microbiota and their implication in anti‐colonic inflammation in mice with colitis induced by dextran sodium sulfate (DSS). Our results showed that DSS treatment dramatically disturbed the composition of gut microbiota in mice, which was associated with elevated colonic inflammation. Dietary treatment with resveratrol partially attenuated the gut microbiota dysbiosis induced by DSS, for example, resveratrol effectively reduced the abundance of bacteria from Verrucomicrobia and Proteobacteria phyla, decreased the abundance of Sutterella and Bilophila genera, and increased the abundance of Bifidobacterium and Lactobacillus genera in DSS‐treated mice. Alpha and beta‐diversity analysis showed that DSS treatment reduced the microbial diversity and shifted the microbial community structure in comparison with the negative control (healthy) mice, while resveratrol ameliorated the aforementioned alteration and restored the diversity of the gut microbiota in DSS‐treated mice. Furthermore, resveratrol alleviated the symptoms of colitis and decreased the expression levels of pro‐inflammatory cytokines such as IL‐10, IL‐2, GM‐CSF, IL‐1β, IL‐6 and TNF‐α, in the colon of DSS‐treated mice. Overall, our data demonstrated that resveratrol partially reversed bacterial dysbiosis in DSS‐treated mice, which may play an important role in its anti‐colitis effect.

Support or Funding Information

This study was partially supported by fund from USDA
Comparison of Cell Growth Inhibition by Various Phenolic‐enriched Sorghum Accessions in Human Liver Carcinoma HepG2 Cells
Authors: Chen Xi , Rhodes Davina , Herald Tom , Su Xiaoyu , Xu Jingwen , Shen Yanting , Wang Weiqun
Published: June 2021

Phenolic agents in plant foods have been associated with chronic disease prevention, especially cancer. In this study, we selected 13 phenolic‐enriched sorghum accessions and tested their extracted phenolic agents on cell growth inhibition in human hepatocellular carcinoma HepG2 cells. Total phenolic contents as determined by Folin‐Ciocalteu were 30–64 mg GAE/g DW on the extracts of various accessions. Treatment of HepG2 cells with the extracted phenolics at 0–200 uM up to 72 hrs resulted in a dose‐ and time‐dependent reduction in cell number. IC50 was varied from 85 to 221 ug/mL for each of the 13 extracted accessions when compared with a negative control at 1,275 ug/mL. Furthermore, the underlying mechanism of cell growth inhibition was examined in one accession, indicating the cell growth inhibition that appeared not to be mediated by cytotoxicity, but cytostatic mechanism with a dose‐dependent increase of cell cycle arrest at G2/M. Meanwhile, the percentage of apoptotic cells were significantly induced. Our on‐going studies conducted on the colon cancer cells will be compared with these liver cancer cell data for a better understanding of anti‐cancer cell activities by sorghum phenolic agents (supported by USDA Cooperative Award 58‐3020‐5‐016).

Support or Funding Information

supported by USDA Cooperative Award 58‐3020‐5‐016
AKT2 is the Predominant Akt Isoform Expressed in Human Skeletal Muscle
Authors: Geddis Alyssa Virginia , Abdalla Mary N , Leandry Luis A , McClung Holly L , Pasiakos Stefan M , Matheny Ronald W
Published: June 2021

AKT is a serine/threonine kinase that mediates a number of intracellular processes including proliferation, survival, and metabolism. Three Akt isoforms have been identified in mammals (AKT1, AKT2, and AKT3), and there exist distinct and redundant functions for each. While a great deal of our knowledge regarding the functions of the three AKT isoforms in skeletal muscle has been derived from the use of cell and animal models, there is limited information regarding the expression and actions of AKT isoforms in human skeletal muscle. Given this, we performed a series of experiments to determine the abundances of each AKT isoform in human skeletal muscle, and to identify the principal AKT isoform involved in human myogenesis. We first examined the RNA transcript expression of each AKT isoform from biopsies obtained from human quadriceps. AKT2 was the most highly expressed AKT transcript, exhibiting a15.5‐fold increase over AKT1 and AKT3 transcripts (P<0.001). No significant difference between expression of AKT1 and AKT3 was observed. We next determined the abundance of AKT protein isoforms by using antibody immunoprecipitation followed by Liquid Chromatography‐Parallel Reaction Monitoring/Mass Spectrometry (LC‐PRM/MS). Immunoprecipitation was performed using both mouse and rabbit pan AKT antibodies that were immunoreactive with purified peptides of all three AKT isoforms. Analysis of immunoprecipitates by LC‐PRM/MS revealed that AKT2 was the most highly expressed AKT in quadriceps muscle (4.2‐fold greater than AKT1 using the rabbit antibody (P<0.001) and 1.6‐fold greater than AKT1 using the mouse antibody(P<0.05)). Expression of AKT3 protein was virtually undetectable in quadriceps, suggesting that AKT3 plays little or no role in skeletal muscle physiology. Having found that AKT2 is the most highly expressed AKT isoform in human quadriceps, we next asked whether AKT2 was the principal isoform regulating human myotube formation. To test this, cultured primary human myoblasts were virally‐transduced with cDNAs encoding either wild‐type (WT) or kinase‐dead (KD) AKT1 or AKT2 and allowed to terminally differentiate. Myotubes expressing either WT‐AKT1 or WT‐AKT2 showed enhanced fusion compared to control myotubes(P<0.05 for each), while myotubes expressing KD‐AKT1 showed a 10% reduced fusion index (P<0.05). However, compared to control myotubes, myotubes expressing KD‐AKT2 showed a 63% decrease in fusion index (P<0.001). In conclusion, our findings identify AKT2 as the predominant AKT isoform expressed in human quadriceps muscle and as the principal AKT isoform that regulates differentiation of cultured human skeletal myoblasts.

Support or Funding Information

The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or reflecting the views of the Army or the Department of Defense.
Transient Receptor Potential Canonical Channel‐1 (TRPC1) KO Mice That Exercise Are Protected from High‐Fat Diet‐induced Obesity and Type 2 Diabetes Risk
Authors: Krout Danielle , Schaar Anne , Singh Brij B , Roemmich James N , Claycombe‐Larson Kate J
Published: June 2021

Objective

Transient receptor potential canonical channel‐1 (TRPC1) is a major class of calcium permeable channels found in key metabolic tissues, including the hypothalamus, adipose tissue, and skeletal muscle, making them likely candidates for the regulation of cellular energy metabolism. The exact role of TRPC1 in development of obesity, adipose tissue mass changes, and obesity‐associated metabolic disease risk has not yet been determined.

Method

Four month old male B6129SF2/J (WT) or TRPC1 knockout (TRPC1 KO) mice were fed a control normal‐fat (NF) (16% fat) or a high‐fat (HF) (45% fat) diet for 3 months and subjected to voluntary wheel running exercise or sedentary activity. Body composition, type 2 diabetes risk, and adipose tissue cellularity were determined.

Results

Under sedentary conditions, body fat mass was not different between WT and TRPC1 KO mice. Fat mass was decreased (P<0.05) in TRPC1 KO mice fed a HF diet and exercised compared to WT mice fed a HF diet and exercised. Fasting glucose concentrations were also lower in TRPC1 KO mice fed a HF diet in both exercise and sedentary groups. Adipocyte numbers were decreased in both the subcutaneous and visceral adipose tissue of TRPC1 KO mice fed a HF diet and exercised.

Conclusion

These findings suggest that in TRPC1 deficient mice, exercise is very protective against HF diet‐induced obesity and insulin resistance. Additional studies will be conducted to determine the mechanism underlying exercise‐mediated reduction in adiposity and insulin resistance in the absence of TRPC1.

Support or Funding Information

This work was supported by the USDA Agricultural Research Service Project #3062‐51000‐052‐00D awarded to K.J.L., the National Institutes of Health awarded to B.B.S. (DE017102; DE024300; GM113123), and by ND EPSCoR through the National Science Foundation (IIA‐1355466) awarded to A.S.
Dietary Prebiotics and Arachidonic Acid (ARA) Modulate Intestinal Injury and Microbial Taxa Following Acute Dextran Sodium Sulfate Induced Colitis in Formula‐Fed Piglets
Authors: Jacobi Sheila K. , Xi Lin , Maltecca Christian , Borst Luke , Smith Andrew , Odle Jack
Published: June 2021

Gastrointestinal (GI) disturbances are among the most common health challenges faced by the suckling neonate, and dietary interventions aimed to modulate inflammation and microbial colonization may provide a prophylactic option to enhance GI development. The objective of this experiment was to investigate the effects of supplemental ARA or prebiotics (alone or combined) on intestinal inflammation and the microbiome in an acute dextran sodium sulfate (DSS) colitis model. Day‐old pigs were randomized among 4 diets (0.5% arachidonic acid (ARA; control); 4g/L galactooligosaccharide + 4 g/L polydextrose (PRE); 2.5% ARA, and PRE+ARA; n=24/diet, and were fed 3 times per d for 21d. On d 17–21 pigs (12 per diet) were treated with 0.625 g DSS/kg BW to induce colitis. Pigs were euthanized on d22 and samples were collected for digesta pH, colonic mucosal fatty acid composition (via GC‐MS), histology, and fecal microbiome analysis (via 16S rDNA Illumina sequencing). Supplements did not affect growth. Colonic mucosal ARA% (w/w) increased from 3.83 in controls to 6.67 in pigs fed diets enriched in ARA (P < .001), with reciprocal reductions in linoleate concentration. Dietary prebiotics increased ARA and linoleate concentrations but reduced stearate concentration in colonic mucosa (P<.002). Colonic mucosal fatty acid composition was unaffected by DSS treatment (P > .05). There was a main effect of PRE on decreasing ileal, cecal, proximal and distal colon digesta pH compared with diets lacking PRE (7.4 vs. 7.3 ± 0.03; 6.2 vs 5.8 ±0.06; 6.3 vs 5.7 ± 0.04; 6.4 vs. 6.0 ± 0.04, respectively; P<0.05). PRE and DSS both increased laxation scores with highest diarrhea being found in pigs fed PRE and challenged with DSS (P<0.05). In addition, pathology assessments (means ± SEM) showed main effects of diet and DSS on epithelial damage, lamina propria inflammation, crypt abscess, crypt dropout, and overall colitis score (P < 0.05). Dietary treatments altered microbial taxa with significant clustering of the PRE diets distinct from ARA diets. In conclusion supplementation of formula with prebiotics significantly shifts microbial taxa on d22, modulates intestinal pH, and laxation.

Support or Funding Information

This project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2014‐67017‐21750 from the USDA National Institute of Food and Agriculture.
Ginger Extracts Inhibit Na‐Glucose Cotransport in Rat Intestinal Epithelial Cells
Authors: Talukder Jamil , Hill Cameron , Jaima Antara , Santiz‐Lopez Jessica
Published: June 2021

Background

Glucose absorption in the intestine is regulated by Na‐dependent glucose transporter (SGLT1). Food borne bioactive compounds may alter the role of Na‐dependent nutrient transport including SGLT1 in the gut lumen. Ginger is one of the common spices being used around the globe and its effect on SGLT1 is unknown.

Aim

To determine the effects of ginger extracts on SGLT1 in enterocytes.

Methods

IEC‐6 cells grown on transwell plates were treated with ginger extracts. Initially, dose response and time course experiments were performed for glucose uptake using [3H]‐O‐M‐Glucose. Cytosolic Ca2+ was measured using Flu 3AM and Caspase‐3 activity was determined using a kit. RTQ‐PCR and Western blot analyses were performed to determine SGLT1 mRNA and protein levels, respectively.

Results

Ginger extracts treatment decreased Na‐dependent glucose uptake dose dependently in IEC‐6. Kinetic studies demonstrated that ginger extracts inhibit glucose uptake by decreasing the affinity of SGLT1. Ginger extracts treatment increased cytosolic Ca2+ levels but not Caspase‐3 activity. Molecular studies showed that SGLT1 mRNA levels and protein on apical membrane were not down regulated by ginger extracts.

Conclusions

These studies revealed that ginger extracts inhibit glucose absorption by inhibiting SGLT1 activity in enterocytes.

Support or Funding Information

This work was supported by NSF grant, HRD‐1332459 to Drs. Golden, Hamada, and Talukder
Newly Formulated Fortified Blended Foods Result in Improved Protein Quality and Iron Bioavailability in Broiler Chickens
Authors: Fiorentino Nicole M , Kimmel Katheryne A , Joseph Michael , Alavi Sajid , Beyer R Scott , Lindshield Brian L
Published: June 2021

Background

Corn and soybean based fortified blended foods (FBFs) have been the primary food aid product provided by the United States. Sorghum and cowpea have been suggested as alternative FBF commodities because they are drought‐tolerant, can be locally grown in areas where food aid is provided, and are not genetically modified.

Objective

To determine protein quality and iron bioavailability of newly formulated sorghum, cowpea, soy, and corn‐based FBFs, compared with the current United States Agency for International Development corn and soy blend FBF, CSB+, in broiler chickens, which have been suggested to be a good model for iron bioavailability.

Methods

Nine groups of one‐day old broiler chicks (n = 10) consumed control chicken diet for one week before being randomized to consume prepared FBFs. The FBFs consisted of corn‐soy blend 14, white sorghum‐soy, white sorghum‐cowpea, white sorghum‐cowpea with soy protein isolate (instead of whey protein concentrate) extruded FBFs, two over‐extruded white sorghum‐cowpea FBFs (one with soy protein isolate), a non‐extruded sorghum‐cowpea FBF, CSB+, or a control chicken diet for 21 days. Results were analyzed by one‐way ANOVA with LSD test using SAS Studio 3.6.

Results

All newly formulated FBFs resulted in significantly higher protein efficiency ratio and caloric efficiency than CSB+. Unlike CSB+, all newly formulated FBFs resulted in significantly increased hepatic iron levels compared to the control group. The white sorghum‐soy group had significantly increased body weight compared to all other FBFs, except the corn‐soy blend 14 group.

Conclusion

Newly formulated FBFs' higher protein quality and iron bioavailability compared to CSB+, suggests they are of higher nutritional quality. Further research is needed to identify the best new FBF formulations.

Support or Funding Information

Supported by USDA‐MFFAPP.
Interrelationships between Inflammatory Biomarkers, Fecal and Plasma Short Chain Fatty Acids
Authors: Rust Bret , Cooper Danielle , Welch Lucas , Horn William , Newman John , Marco Maria , Martin Roy , Keim Nancy
Published: June 2021

Short chain fatty acids (SCFA) have been associated with improved glycemia and to reductions in inflammatory responses in cell and rodent models. Low grade inflammation has been associated with insulin resistance. We measured fecal and plasma SCFA and inflammatory biomarkers in response to a proprietary oligosaccharide intervention. 22 overweight and obese men and women participated in a three‐way crossover study that included 3 levels of oligosaccharide: 0, 15 and 30 g/d. for 3 weeks. During each treatment, a collection of 5 days' feces were combined and analyzed for SCFA. At the end of each treatment, a meal challenge protocol was employed in which participants consumed half their daily dose of oligosaccharide with breakfast followed by a standard lunch. We measured plasma acetate, propionate and butyrate in 12 blood draws over the course of a 9 hour test day and inflammatory cytokines at 3 time points, 0, 195 and 435 min following the breakfast test meal. All SCFA analyses were conducted by GC‐MS. Inflammatory cytokines were measured on an 8‐plex chemiluminescent MSD plate except for adiponectin and CRP which employed an Integra 400 bio‐analyzer. Breath hydrogen and methane were collected to characterize participants as either methane producers (MP) or non‐producers (NM). Adiponectin was positively associated with fecal SCFA (p<0.05). TNF‐α and IL‐10 were negatively associated with various plasma SCFA AUC segments (p<0.05). Neither CRP nor IL‐6 was associated with either fecal or plasma SCFA. Fecal SCFA concentrations did not correlate well with plasma SCFA AUCs. Fecal propionate and butyrate were lower in MP than NM. Breath methane was negatively associated with SCFA production in MP. Plasma and fecal SCFA may predict some inflammatory responses but differences in the time of collections and the span of collections may reflect different temporal relationships to cytokines.

Support or Funding Information

Funding provided by USDA CRIS 2032‐51530‐022‐OOD and the West Coast Metabolomics Center (U24 DK097154)
Pig Has No Brown Adipose Tissue
Authors: Hou Lianjie , Hu Ching Yuan , Wang Chong
Published: June 2021

Brown adipose tissue (BAT) is critical for mammal's survival in the cold environment. UCP1 is responsible for the thermogenesis in the BAT and is recognized as a specific marker for BAT. Pig is a useful medical model for humans due to its similarity in size and physiology. Pig is also important economically as a meat‐producing livestock. A previous study identified brown adipocytes in different pig adipose tissues by microscopic and electron microscopic methods. In one immunoblotting study, UCP1 was not detected in any porcine adipose tissues by the rabbit anti‐rat UCP1 antibody. But, a recent study showed that UCP1 protein is present in porcine adipose tissue and is responsive to postnatal leptin treatment. These authors used an ovine UCP1 antibody in their immunoblotting assay. However, the positive result may be explained by non‐specific hybridization with other mitochondrial anion carrier proteins, such as UCP2 or UCP3 by the antibody used in the previous study. Therefore, whether BAT or more precisely UCP1 protein, exists in pig has never been conclusively established. The objective of this study was to ascertain whether pig has UCP1 protein.

In this study, we used rapid amplification of cDNA ends (RACE) technique to obtain the UCP1 mRNA 3′ end sequence, confirmed only exons 1 and 2 of the UCP1 gene are transcribed in the pig. We then cloned the pig UCP1 gene exons 1 and 2, and used these two exons to express pig UCP1 protein by E. coli BL21. We purified and used the expressed pig UCP1 protein as antigen for antibody production in a rabbit. Immunoblotting assay was conducted using inguinal fat, interscapular fat, epididymal fat, retroperitoneal fat, supraclavicular fat, perirenal fat, neck fat, liver, muscle from pigs. Mice BAT was used as a positive control. Our rabbit anti‐pig UCP1 antibody can hybridize with mice UCP1 protein (about 35KD) and pig UCP1 (about 15KD) expressed by E. coli BL21, but could not detect UCP1 protein in any pig tissues. Our data clearly show only exons 1 and 2 of UCP1 gene are transcribed in pig adipose tissue, but the truncated pig UCP1 mRNA is not being translated. Since these results unequivocally demonstrate that pig has no UCP1 protein; therefore, the pig has no brown adipose tissue.

Support or Funding Information

This work was supported by the National High Technology Research and Development Program 863 (#2013AA102502); the National Natural Science Foundation of China (#31372283); The Team Project of Guangdong Agricultural Bureau (#2016LM2148) and the Natural Fund Key Projects of Guangdong Province (#2015A030311006). C.H. is supported by the USDA National Institute of Food and Agriculture, Hatch project HAW‐H2037, managed by the College of Tropical Agriculture and Human Resources, University of Hawaii at Manoa.