bluuebunny/crossref_metadata_2025 huggingface dataset:
UV irradiation/cold shock‐induced NOS2 expression for causing nuclear bubbling is WWOX and p53 dependent (1010.11)
Authors: Chang Nan‐Shan , Chen Szu‐Jung , Lin Pei‐Wen
Published: June 2021

When a cell receives UV irradiation and cold shock, an untypical apoptosis occurs rapidly in less than 5‐20 min. We determined here that the UV energy activates cytosolic NOS2 (Nitric oxide synthase 2) for generating nitric oxide (NO)‐containing gas, which causes nuclear pore enlargement and a single large‐size bubble formation, as determined by electron microscopy (thus designated bubbling). Unlike typical apoptosis, no DNA fragmentation, nuclear condensation, membrane blebbing, and flip over of phosphatidylserine (PS) on cell membrane was observed. UV irradiation activated tumor suppressor p53 with phosphorylation at Ser46 and WWOX at Tyr33. Intriguingly, cold shock, but not UV, activates antiapoptotic TRAF2 (TNF‐α receptor‐associated factor 2) and downstream NFκB (nuclear factor κB). TRAF2 is abundant in the normal skin, and is significantly decreased in frostbitten skin tissues. In contrast, the expression of tumor suppressor WWOX is greatly upregulated in frostbitten skin. Mechanistically, UV/cold shock induced WWOX release from the WWOX/TRAF2 complex in 30 minutes, and the released WWOX bound p53 for relocating to the nucleus and caused cell death. Both dominant negative p53 and WWOX abolished the effect of UV/cold shock. Also, the UV/cold shock effect was blocked in Wwox‐/‐ MEF cells and in cells transiently overexpressed with TRAF2. Taken together, UV energy activates cellular NOS2 to generate nitric oxide‐containing gas in the nucleus, and causes cell damage in a WWOX/p53‐dependent manner. (Research supported by NSC and NHRI, Taiwan, and DoD, USA)

Grant Funding Source: national science council; national health research institute
Synthesis and anti‐microbial studies of a series of 3‐formylchromone thiosemicarbazone ligands and their Cu(II) complexes including the potent Akt inhibitor [Cu(FC‐TSC)Cl2] (1010.15)
Authors: Werlein Amanda , Lisic Edward
Published: June 2021

A series of six thiosemicarbazone compounds synthesized from a 3‐formylchromone backbone and the synthesis of their copper (II) complexes is reported here. Each of the synthesized thiosemicarbazone ligands and their corresponding copper (II) complexes were tested for antimicrobial activity by determining the minimum inhibitory concentrations (MIC) against two gram negative bacteria (Escherichia coli and Pseudomonas aeruginosa), two gram positive bacteria (Bacillus subtilis, Staphylococcus aureus,), two yeast (Candida albicans and Saccharomyces cervisiae), and one mold (Aspergillus niger). The series of 3‐formylchromone thiosemicarbazone ligands showed little to no anti‐proliferative activity. However, the copper (II) complexes of the ligands, including the potent Akt inhibitor [Cu(FC‐TSC)Cl2], have somewhat better anti‐proliferative activity especially against the gram positive bacteria.
Anti‐apoptotic mechanism(s) of angiotensin 1‐7/mas in alveolar epithelial cell survival (1010.3)
Authors: Gopallawa Indiwari , Uhal Bruce
Published: June 2021

Earlier work showed that autocrine synthesis of angiotensin II (ANGII) and c‐Jun‐N‐terminal kinase phosphorylation (pJNK) are required events in alveolar epithelial cell (AEC) apoptosis which is linked to severe lung diseases. Angiotensin 1‐7 (ANG 1‐7) prevents AEC apoptosis which is induced by locally synthesized ANG II. However the exact inhibitory mechanisms of ANG 1‐7/mas are currently unclear in AECs. The objective of the current research is to identify molecular mechanism(s) by which ANG1‐7 promotes AEC survival. We hypothesized that ANG 1‐7 activates a map kinase phosphatase (MKP) and maintains low pJNK levels thereby preventing apoptosis. To evaluate whether adding ANG 1‐7 acts through MKP‐2, freshly prepared ANG 1‐7 (0.1µM) in serum free media was added to a mouse lung epithelial cell line (MLE‐12). Cell lysates were run on 10% SDS‐PAGE to detect MKP‐2 protein by Western blotting. Current preliminary data suggests that ANG 1‐7 increased MKP‐2 protein (p=0.0286). Further, blockade of endogenous ANG 1‐7 with the mas receptor antagonist A779 reduced MKP‐2 levels (p=0.0014). Moreover data shows that inhibition of protein kinase A (PKA) in MLE‐12s induced pJNK and hence may be involved in promoting AEC survival. The results from these studies suggest that MKP‐2 is a key protein in the ANG 1‐7/mas pathway, and may provide insights to potential therapeutic strategies for lung diseases that involve AEC apoptosis.

Grant Funding Source: Supported by HL‐45136
Expression in yeast of the metacaspase scp3 from the fungus Schizophyllum commune (1010.8)
Authors: Kaminski Anne , Fox Kristin
Published: June 2021

Caspase homologs, metacaspases, are found in fungi, protists, and protozoa and are involved in programmed cell death‐like behavior. There is also evidence that metacaspases are additionally involved in proteostasis and cell cycle regulation. Metacaspases contain a cysteine histidine catalytic diad and cleave after lysine and arginine. Some metacaspases are activated by calcium and autocatalytic processing occurs in several, but is not necessary for activity.

Five metacaspases, named scp1‐5, have been identified from the fungus Schizophyllum commune with the goal of expressing and characterizing them. Previous attempts to express these metacaspases in E. coli have resulted in either low amounts of protein or protein that forms inclusion bodies. The yeast Saccharomyces cerevisiae is often used to express eukaryotic proteins and should allow the production of large amounts of soluble protein. Therefore, the expression of these metacaspases in yeast is being investigated as an alternative to expression in E. coli.

The scp3 gene has been successfully cloned from S. commune and inserted into the vector pYES2.1/V5‐His‐TOPO (Life Technologies). The vector containing scp3 was transformed into S. cerevisiae. Expression has been attempted by induction with galactose through the GAL1 promoter in the vector. Scp3 will be detected by western blotting using a V5 antibody (Invitrogen).
Effects of heparin and heparin‐binding growth factor on human aortic adventitial fibroblasts (1012.4)
Authors: Kalle Fanta , Akins Robert
Published: June 2021

Cardiovascular diseases are primary treated using bypass graft surgery and angioplasty; unfortunately, these procedures often fail due to maladaptive tissue remodeling leading to stenosis, fibrosis, and vessel failure. Methods to decrease failure rates are needed. We are developing instructive biomaterials for placement along the abluminal surface of at‐risk vessels to provide mechanical support and to deliver bioactive molecules and/or stem cells to help attenuate maladaptive responses, encourage healing, and improve clinical outcomes. The present study examines the effects of critical biomaterial components including basic fibroblast growth factor (FGF2), low‐molecular weight heparin (LMWH) and desulfated LMWH on vascular cells.

Cell morphology, proliferation, and viability are assessed using fluorescence microscopy. Image Pro and SPSS software packages are used to acquire and analyze data. Data indicate that FGF2 and LMWH affect human aortic adventitial fibroblast (AoAF) phenotype. The presence of FGF2 significantly increased proliferation rates and the appearance of filopodia; whereas, the absence of FGF2 was associated with an increase in the prevalence of lamellipodia and stress fibers. Increasing LMWH doses correlated with decreased AoAF proliferation and alterations in cell morphology. Desulfated LMWH did not alter FGF2‐mediated effects on cell phenotype.

These data suggest that the biomaterial components LMWH and FGF2 can influence the phenotype of cardiovascular cells like AoAFs. Although in vivo, LMWH is thought to potentiate FGF2‐induced effects, in our system, LMWH significantly decreased AoAF proliferation, particularly in the absence of FGF2. The sulfation state of LMWH was critical to these effects. Further studies to determine the mechanisms accounting for these effects and to establish combined effects on cell phenotype will be needed to enable development of injectable formulations that may be useful in improving clinical outcome.

Grant Funding Source: supported by the Delaware INBRE program with a grant from the National Institute of General Medical
Applying a chemical genetic approach to inositol polyphosphate signaling (1013.10)
Authors: Bratz Jeffrey , Dul Barbara , Fiedler Dorothea
Published: June 2021

Signal transduction is a key area of study as we try to understand habitual cell physiology, and also discover new targets for amelioration of disease symptoms. Inositol polyphosphates, IPs, are small molecule messengers that play a critical role in cellular signal transduction pathways.

Several highly phosphorylated IPs, such as 5‐diphosphoinositol pentakisphosphate, (5‐PP‐IP5) have been linked to insulin signaling in mammalian cells and maintaining overall energy balance in yeast. While these phenotypes are exciting, the field has been limited by traditional genetic techniques, and the exact mechanism of action by IPs has remained elusive.

Through the development of a chemical genetic technique largely applied to protein kinases, we have engineered analog sensitive kinases responsible for the metabolism of IPs in yeast. We have exhibited the functionality of these enzymes, while simultaneously displaying the ability to specifically and temporally inhibit their kinase activity in both in‐vitro and in‐vivo assays. We plan to use this system in conjunction with metabolomics to further elucidate the role that these intriguing small molecules play in signal transduction pathways.

Grant Funding Source: NIH
Nucleotide regulation of soluble guanylate cyclase (1013.14)
Authors: Surmeli Nur , Marletta Michael
Published: June 2021

The interaction of the enzyme soluble guanylate cyclase (sGC) with nitric oxide (NO) mediates numerous physiological and pathological processes. NO production regulates smooth muscle relaxation, myocardial function, perfusion and platelet aggregation. NO signaling engages the second messenger cyclic guanosine monophosphate (cGMP) synthesized by sGC. Due to the critical importance of cGMP in many cellular processes, sGC activity is tightly regulated by NO, ATP and GTP. The mechanistic details of sGC regulation by these molecules are not well understood. Nucleotides influence sGC activity via a poorly characterized allosteric site. The catalytic domain of sGC has two putative nucleotide binding sites. One of these sites lacks an aspartate residue that is essential for catalysis, and is denoted as the pseudosymmetric site. The role of pseudosymmetric site on nucleotide regulation was investigated by site‐point mutations at this site. Kinetic analysis of the pseudosymmetric site mutant at various NO states has shown that pseudosymmetric site is involved in nucleotide regulation of sGC, and the effect of nucleotides is also dependent on the NO binding state of the enzyme.

Grant Funding Source: American Heart Association
Antitumor activity of sphingosine‐1‐phosphate (1013.20)
Authors: Yang Jian , Ling Binbing , Sultan Ahlam , Michel Deborah , Zhang Huihua , Ma Baohua , Maley Jason , Sammynaiken Ramaswami , Alcorn Jane
Published: June 2021

Sphingosine‐1‐phosphate (S1P) is an endogenous metabolite derived from ceramide as part of the sphingomyelin cycle. It is a potent molecular messenger which exerts its function both intracellularly and extracellularly. Intracellularly, S1P acts as a second messenger regulating calcium mobilization, and cellular proliferation and survival. Extracellularly, S1P acts as a ligand of the G‐protein‐coupled S1P receptors (S1PRs) and mediates a variety of physiological and pathological processes. Levels of S1P and other sphingosine metabolites are maintained in a delicate balance through the action of two enzymes, sphingosine kinase (SK) and sphingosine‐1‐phosphate lyase (SPL). Our previous studies showed that exogenously administered S1P (concentration: 1 µM) exhibited synergistic effects with chemotherapy drugs in human breast cancer MCF7 and MDA‐MB‐361 cells [1, 2]. Herein, we report other cell lines and mouse xenograf study results on the antitumor activity of S1P.
Factors influencing the decision to formula feed infants (1016.2)
Authors: Brown Nicole , Gregoire Mary , Sowa Diane , Chen Yimin , Ratz Natalie
Published: June 2021

This research investigated factors that contributed to 88 mothers’ decisions to formula feed (either exclusively or in conjunction with breast milk) infants delivered at a Midwestern academic medical center where many Baby Friendly practices are used. The sample of black (39%), Hispanic (39%), and white (17%) mothers had a mean age of 27.8 years; 35.2% were married. The mother’s decision to formula feed was consistent with how the infant’s father thought the baby should be fed (73% preferred formula) for white mothers, less so with black (48% of fathers preferred formula) and Hispanic mothers (35%). The opinions of the baby’s father, mother’s mother, and mother’s friends on how the baby should be fed were of somewhat limited importance to all mothers regardless of race (m<3.5 on 5 pt scale). Mothers’ scores on the Iowa Infant Feeding Attitude Scale were similar across race (scores 56, 56, 57 out of 85 with lower scores indicating stronger attitudes towards formula feeding). Few of the mothers (<20%) indicated that their doctor, their baby’s doctor, or the hospital staff favored their choice of formula feeding their infant and only a third indicated they received help with formula feeding while in the hospital. Mothers choosing to formula feed in a hospital focusing on Baby Friendly practices may receive limited support from physicians and staff.
Breastfeeding intentions and practices among the different vegetarian groups in the United States (1016.8)
Authors: Armstrong Anne , Anderson Alex
Published: June 2021

The study assessed the breastfeeding (BF) intentions and practices among the different types of vegetarian women in the United States. This was a cross‐sectional online survey. The survey was emailed to administrators of vegetarian listservs for the membership to complete. Most of the respondents were vegans (38.9%), Caucasians (77%), employed (80.3%), married (53.4%), and have education above high school (82.1%). 74.5% were breastfed as a child. Overall, the proportion of participants who intended to BF was higher than actual BF practice, irrespective of vegetarian type. While intentions for exclusive breastfeeding (EBF) for 6 months was 67.7%, 51.7% of the participants reported to have EBF a previous child for 6 months. While 50% of flexitarians intended to EBF, 79.1% of vegans intended to EBF for 6 months. BF practice was generally low compared to BF intentions among the different vegetarian groups with only 5.1% of flexitarians, lacto‐vegetarians and ovo‐vegetarians, respectively compared to 44.4% vegans reported to have BF a previous child. In conclusion, although a greater percentage of vegetarians intend to BF, only a small proportion actually reported to have BF, suggesting a disconnect between BF intentions and actual practice. Our findings suggest an understanding of the barriers to BF among the different vegetarian groups to inform effective BF promotion and support for these groups of women.
Dynamics of dining in the groups of people (1019.17)
Authors: Sigirci Ozge , Wansink Brian
Published: June 2021

Objective: In an all‐you‐can‐eat (AYCE) buffet, how does diners’ eating and taste orientations change based on the gender of people they are dining with? Methods: The consumption amounts and taste evaluations of diners eating at an AYCE restaurant were taken along with other measures of behavior and self‐perceptions. The differences between males sitting in groups of females and males sitting in groups of males vs. females sitting in groups of males and females sitting in groups of females are analyzed using T‐Test.

Results: Males who dine with females eat more pizza and salad than males who dine with males (p<0.05); on the other hand, females who dine with males eat more pizza but less salad than females who dine with females. Males dining with females find the meal and pizza tastier and more enjoyable than males dining with males, whereas females dining with males find the meal and pizza less tasty and less enjoyable than females dining with females. Moreover, females dining with males think they ate more than they should have and felt rushed than females dining with females (p<0.05).

Conclusions: Dining in a group of people with same sex vs. mixed sex influences diners’ eating behaviors. Although eating with females lead males to eat more, they also eat more of healthy options such as salad and enjoy the meal more. But eating with males lead females to eat more and choose more unhealthy options, and enjoy the dinner less. A diner wanting a more enjoyable and perhaps a healthier experience might want to realize that eating in groups and eating with males or females will affect and alter the eating behavior.
Evaluation of the effectiveness of a cafeteria‐based behavioral economics intervention designed to improve student intake of reimbursable meal components (1019.18)
Authors: Golub Haleigh , Gunther Carolyn , Kennel Julie
Published: June 2021

Behavioral economics (BE), the study of the effects of psychological factors on decision‐making, can be used in school cafeterias to influence student food choice and consumption. This study aimed to determine the impact of a cafeteria‐based BE intervention on student intake of meal components (entrée (grain/protein), vegetable, fruit, milk) and associated key nutrients (eg, fiber). Based on an assessment of cafeteria environments using The Smarter Lunchrooms Movement protocol, a BE intervention (identical across sites) was implemented in 4 elementary schools. A process evaluation checklist measured fidelity of intervention. Plate waste, an indirect indicator of dietary intake, was estimated 3 times before and after the intervention by scoring each meal component on a 5‐point scale from 0 (all item eaten) to 4 (all item wasted). Estimated nutrient intakes were calculated by comparing plate waste data against nutrient content of food items per serving size. Of the 637 students’ trays surveyed from the 4 schools at baseline, 21.8% and 44.5% of students ate all or none of the entrée, respectively; 68.7% ate less than 1/4 of the vegetable; and 60.3% ate 3/4 or all of the fruit. Only 13.7% of students chose unflavored milk over flavored varieties. Findings from this study will determine the effectiveness of a BE intervention designed to increase student intake of reimbursable meal components and key nutrients in schools.

Grant Funding Source: Supported by the Ohio Department of Education Team Nutrition grant
Purchasing behaviors of individuals who shop at corner stores (1019.6)
Authors: Craven Tara , Swanson Ann , Young Taylor , Martin Donna , Gibson Cheryl
Published: June 2021

Background. The Jackson County MO Healthy Corner Store Initiative is designed to work with small retail establishments to increase healthy food availability where residents have limited access to full line grocery stores. As part of this initiative, we assessed the purchasing habits of individuals who frequent one of four neighborhood corner stores. Method. Intercept surveys were administered during 28 two‐hour sessions and 2 four‐hour sessions. Survey sessions were held early morning, mid‐day and late afternoon over a 2‐week period. Participants were asked to indicate age, gender, transportation mode, food purchase frequency, typical items purchased, perceived healthiness of food items available, what influenced purchase decisions, and what items they purchased that day. Results. 446 surveys were completed. Most participants were greater than 18 years old (88%), male (61%) and drove a car or truck to the store. About 36% reported visiting corner stores daily. Most participants purchased snack items (59%) and felt that available food items were somewhat healthy (38%). Chips (17%), regular soda (25%), alcohol (15%) and tobacco (14%) were common items purchased. Almost 50% of participants reported to be very interested in healthier food options while about 23% felt that prices of healthy food items were comparable to other stores. 44% indicated variety would be the most important factor when deciding to purchase healthy foods. Discussion. Intercept survey results provided insight about buying habits and what changes might be possible to increase healthier food purchases. Education about healthy food choices will be an important factor when implementing the Healthy Corner Store Initiative.

Grant Funding Source: Supported by CDC
Development of a food preference instrument for energy dense foods and sugar sweetened beverages (1022.2)
Authors: McGuerty Amber , Cater Melissa , Prinyawiwatkul Witoon , Tuuri Georgianna
Published: June 2021

The current environment may promote adolescent palate preferences for highly marketed, energy dense foods and sugar‐sweetened beverages. The present study developed and examined the construct validity of a food preference survey to estimate adolescent’s willingness to eat these items. Five focus group interviews with 13 to 19 year‐old male and female high school students were conducted. A list of 5 items representing each of the following was developed: energy‐dense foods, energy‐dilute foods, sweetened beverages, and unsweetened beverages. The final survey instrument was administered to 234 public high school students. Willingness to consume each of the 20 items was assessed using a 7‐point Likert Scale with 1 representing “Extremely Unwilling” to 7 indicating “Extremely Willing.” Exploratory Factor Analysis using Principal Axis factoring with a Promax (oblique) rotation found two factors. Factor one included French fries, Kool‐Aid, glazed donuts, cookies, lemonade, and pizza (23.9% of the variance). Factor 2 included nuts or peanut butter, low‐fat or fat‐free yogurt, grapes, and bananas (13.8% of the variance). Internal consistency measured by Cronbach’s alpha was 0.770 for factor 1 and 0.664 for factor 2. The factor explaining most of the variance included foods processed with added fat and/or sugar.

Grant Funding Source: Supported in part by Hatch Act Funds # LAB 93846
Soy intake and age at onset of menarche among adolescents with a high and wide range of soy consumption (1024.1)
Authors: Sabaté Joan , Siapco Gina , Pribis Peter , Oda Keiji
Published: June 2021

Early onset of menarche impacts the future health of adolescent girls. We examined if soy is related to age at onset of menarche (AOM) in a population with a high and wide range of soy intake. We conducted a cross‐sectional study on 339 girls ages 12‐18 years in selected middle and high schools in California and Michigan using a web‐based physical development and dietary intake (DI) questionnaire. Soy consumption (categorized as total soy, meat analogs, tofu/ traditional soy, and soy beverages) was estimated from the DI while AOM was self‐reported. Data analyses included descriptive statistics, Cox proportional hazard ratios, and Poisson regression with adjustment for confounders. Mean (SD) intakes were: total soy,12.9 (14.4) svg/wk; meat analogs, 7.0 (8.9) svg/wk; tofu/traditional soy, 2.1 (3.8) svg/wk; soy beverages, 3.8 (6.3) svg/wk. Mean AOM for those who reached menarche was 12.5 (1.4) y. Consumption of total soy and the 3 types of soy foods were not significantly associated with AOM neither with the odds for early‐ or late‐AOM. Adjustment for demographic and dietary factors did not change the results. Soy intake is not associated with AOM in a population of adolescents who have a wide range and relatively higher intake of soy than the general US population. Our finding suggests that the increasing popularity of soy in the US may not be associated with the declining trend in menarcheal age.

Grant Funding Source: WhiteWave
Adiponectin receptor 1 mediates regulation of bone formation and osteoblast differentiation by GSK‐3β/β‐catenin signaling (1027.1)
Authors: Lin Yuan Yu , Chen Ching Yi , Wu Shinn Chih , Mersmann Harry , Ding Shih Torng
Published: June 2021

The purpose of this study was to evaluate the role of adiponectin receptor 1 (adipoR1) on bone physiology using porcine AdipoR1 transgenic mice (pAdipoR1) as a model. We found that pAdipoR1 transgenic mice have higher bone mineral density than wild‐type mice in older ages. The percent bone volume and trabecular number determined by micro‐CT, was also significantly higher in pAdipoR1 transgenic mice than in wild‐type female mice at both young and older ages. ELISA analysis revealed that serum osteocalcin (Oc) and osteoprotegrin (OPG) were significantly increased in young pAdipoR1 transgenic mice in both genders. Furthermore, serum OPG also elevated at 8 week and 32 week in female and male pAdipoR1 transgenic mice at young and old ages. Serum TRAP5b concentration was reduced in male pAdipoR1 mice compared with wild‐type mice. Knock‐down AdipoR1 significantly decreased the gene expression of Oc, OPG, alkaline phosphatase and msh homeobox 2 and mineralization in MC3T3‐E1 and mesenchymal stem cells, suggesting that AdipoR1 may mediate the function of bone growth. In addition, we found that AdipoR1 may regulate osteoblasts differentiation through GSK‐3β/β‐catenin signaling by pathscan analysis. We conclude that adipoR1 is a critical factor regulating osteoblast differentiation and bone homeostasis.
Effects of defatted dried bonito fish on bone mass, bone composition, and bone strength in Wistar and Goto‐Kakizaki rats (1027.3)
Authors: Ochiai Masaru , Gohtani Shoichi , Matsuo Tatsuhiro
Published: June 2021

Obese type 2 Diabetes (T2DB) is well known to induce osteoporosis in older animals, but little is known about bone health in non‐obese T2DB growing animals. Dietary vegetable proteins and polyunsaturated fatty acids, especially eicosapentaenoic and docosahexaenoic acids, are effective to enhance bone health in aged rats and ovariectomized rats, but the bone developing effects of fish proteins have not been investigated in non‐obese T2DB growing animals. Dried bonito fish (BF) is one of the important and traditional food materials in Japan, but its nutritional perspectives are not well known. In this study, we prepared the 97% defatted dried bonito fish (DBF) using petroleum ether and hexane from BF and then investigated the effects of dietary DBF on bone strength, bone mass, and bone composition in Wistar (WT) and non‐obese T2DB Goto‐Kakizaki (GK) rats. WT and GK rats aged 5 weeks were divided into three dietary groups: casein, BF, and DBF, and then fed each diet for 6 weeks. At the end of the treatment period, the serum and femur were collected for analysis. Dietary BF and DBF enhanced maximum load, structural stiffness, and dry weight of the femur in both WT and GK rats. In GK rat study, dietary DBF decreased the serum glucose level and alkaline phosphatase activity in parallel with a little improvement of femur strength. However, remarkable differences in bone health were not observed between the BF and DBF groups in both rats, which indicated that the main active nutrients to enhance bone health were not lipids but proteins which contained in BF. These findings conclude that dietary proteins derived from BF and DBF can enhance bone health in WT and non‐obese T2DB GK rats.
High refined carbohydrates diet promotes comparable hepatic tumorigenesis in mice as high fat diet, potentially through inducing endoplasmic reticulum stress PKR‐like kinase signaling (1029.1)
Authors: IP Blanche , Liu Chun , Smith Donald , Wang Xiang‐Dong
Published: June 2021

Previous animal studies showed that obesity induced with a semi‐purified high fat diet (HFD) promoted hepatocellular carcinoma (HCC) development as compared to mice fed a non‐semi‐purified chow, through inducing systemic and hepatic inflammation. Since ingredients in chow may elicit chemopreventative effects that are absent in HFD, this present study evaluated the liver tumorigenic effects of dietary fat by replacing it with refined carbohydrates (digestible saccharides; HCD), without altering other dietary components. C57Bl/6J mice fed ad‐libitum HCD (66% energy from carbohydrates) for 24 weeks resulted in comparable degree of carcinogen‐initiated liver tumorigenesis (tumor number and volume) as HFD‐fed mice (60% energy from fat), despite having reduced body weights, suggesting HCD may promote HCC through obesity‐independent pathways. As compared to HFD, HCD: 1) induced liver endoplasmic reticulum (ER) stress‐mediated PKR‐like kinase (PERK) activation; 2) increased hepatic oncogenic IL‐6/STAT3 signaling; 3) induced Akt‐Erk1/2 activation and cyclin D1 protein expression in tumors; 4) promoted pro‐apoptotic CHOP and cleaved caspase‐3 protein expression in tumors; 5) had more severe liver steatosis; 6) elevated hepatic de novo lipogenesis (ACC, SCD gene and protein). This data indicated that chronic HCD consumption can result in comparable severity of hepatic tumorigenesis as HFD, potentially through inducing ER stress‐mediated PERK signaling.

Grant Funding Source: USDA/ARS grant 1950‐51000‐074S
A case study of the influence of maternal pregnancy body composition changes on newborn outcomes (1031.16)
Authors: Weekley Arielle , Harper Kristin , Bailey Caitlin , Anderson Alex
Published: June 2021

Studies suggest women are entering pregnancy overweight/obese and gaining excess gestational weight, both negatively associated with pregnancy outcome. This was a case study that examined the impact of gestational body composition changes of 3 pregnant women and their newborns. Maternal body composition was monitored via the BOP POD at 16, 24 and 36 weeks gestation as well as 2 weeks postpartum. Newborn body composition via the PEA POD was measured at 2 weeks after delivery. All 3 mothers were Caucasians, aged 29 yrs, 34 yrs and 37 yrs, pre‐pregnancy BMI of 18.8 kg/m2, 22.1 kg/m2 and 40.5 kg/m2, respectively. All cases have college level or more education. Pregnancy weight gain was within the recommended gestational weight gain for each case. Interestingly, the obese case lost percent body fat but gained fat free mass during gestational weight gain. Although, both the normal weight and lean cases gained percent body fat during pregnancy, the gain was lower in the normal weight case compared to the lean case (4.2% vs. 7.9%). Overall, each case had normal vaginal deliveries of normal birth weight newborns. The infant of the normal weight case weighed the most (3.38 kg) at birth while that of the lean case weighed the least (3.24 kg). Findings from this study suggest that percent body fat gained during pregnancy may be associated with newborn adiposity and not the absolute gestational weight gain.

Grant Funding Source: Hatch‐GEO00701
Cannabinoid receptor expression in femora and tibiae of C57/blk6 mice fed DHA and relationship to bone ash and BMC (1032.2)
Authors: Kim Jeffrey , Insogna Karl , Watkins Bruce
Published: June 2021

Although the endocannabinoid system (ECS) acts in most peripheral tissues its biology remains largely unexplored in organs besides the brain. The ECS utilizes cannabinoid receptors, CB1 and CB2, and lipid ligands derived from polyunsaturated fatty acids (PUFA) including the n‐3 PUFA. Given the current epidemic of osteoporosis we sought to examine the role of the ECS in bone. Male C57/blk6 mice were fed a semi‐purified diet (AIN 93M) with either DHA (n‐3 PUFA) containing lipid or control lipid for 118 d to assess changes in growth and skeletal metabolism. No difference was observed in body weight during the study. Mice femora and tibiae were collected at 62 and 118 d of the feeding study. DXA analysis showed a higher BMC per body weight in mice after 62 d of DHA feeding compared to the controls. Femora and tibiae ash weight was higher in the DHA fed mice at 118 d. At day 118, mice on both diets were found to have higher expression of CB1 than CB2 in subarticular chondrocytes in the femur and to a lesser extent in tibiae. In trabecular bone, DHA fed mice had a higher level of CB2 expression compared to controls. It is noteworthy that CB2 has been previously shown to promote osteoblast bone remodeling. Taken together, these findings demonstrate that DHA diet fed mice have a higher BMC per body weight and femora/tibiae ash that may be associated to a change in the skeletal topography of the ECS receptors.
Ripening of cheddar cheese with Lactobacillus helveticus as adjunct culture produces peptides that protect human vein endothelial cells against inflammatory markers relevant to cardiovascular disease (1034.12)
Authors: Ravisankar Shreeya , Yuan Jialing , Munske Gerhard , Noratto Giuliana
Published: June 2021

Lactobacillus helveticus has been recognized as an adjunct culture that contributes to flavor development in cheddar cheese. However changes in peptides profiles throughout ripening and their anti‐inflammatory effects on HUVEC have not been investigated.

Peptides in cheddar cheese (Cougar Gold) were assessed by LC‐MS/MS analysis throughout ripening (0, 6, and 12 months). The protective effects of peptides in HUVEC challenged with glucose (G) or E. coli lipopolysaccharides (LPS) were assessed by quantification of reactive oxygen species (ROS), H2O2, glutathione S‐transferase activity (GST), and expression of inflammatory genes by standard analytical methods.

Results showed that peptides produced within 0‐12 months ripening varied in masses from 1226 to 7474 Da with the lowest masses found at 12 months. These peptides prevented ROS production in HUVEC challenged with LPS or G to 0.7‐fold and to 0.36‐fold of LPS‐ and G‐controls, respectively and this was correlated with levels of H2O2. These effects were in part due to higher induction of GST by the 12 month peptides (1.64‐fold of control) and downregulation of NF‐ĸB mRNA and NF‐ĸB‐target genes IL‐8, IL‐6, IL‐1β, and ICAM1. We are currently investigating the peptide profiles and bioavailability throughout ripening. Overall, these results strongly suggest that these peptides can protect HUVEC against inflammation and the mechanisms involve more than the widely reported antihypertensive activities with relevance for CVD prevention.
Role for dusp4 in modulating PPARγ activity by covalent modification in adipocytes (1037.12)
Authors: Guo Wei , Ferguson Bradley , Nam Heesun , Hopkins Robin , Morrison Ron
Published: June 2021

Peroxisome proliferator‐activated receptor gamma (PPARγ) is a ligand‐activated transcription factor that plays a prominent role in adipocyte differentiation and insulin sensitivity with aberrant activity linked to obesity and type 2 diabetes. Recent studies have highlighted covalent modification of PPARγ, including phosphorylation, sumoylation and ubiquitylation, as an important mechanisms modulating biological outcome. Phosphorylation of PPARγ by mitogen‐activated protein kinases (MAPKs; ERK, JNK) inhibits transcriptional activity while mice with targeted mutation against the single MAPK‐consensus recognition site are protected against diet‐induced insulin resistance. Others have also shown MAPK‐dependent phosphorylation of PPARγ as an obligatory step in sumoylation‐dependent inhibition of activity. As MAPK activity is modulated by kinases and phosphatases, the objective of this study was to investigate a role for MAPK‐specific phosphatases in modulating PPARγ activity through covalent modification. We examined the expression profile of the 10 known MAPK‐specific phosphatases, collectively referred to as dual‐specificity phosphatases (DUSPs), in insulin‐responsive tissues of genetic‐ and diet‐induced obese mice. We observed that dusp4 was significantly more abundant in white adipose tissue relative to liver, skeletal muscle, and heart in lean mice and that this phosphatase was induced during early stages of obesity and repressed during later stages correlating with the onset of obesity‐induced inflammation. The expression profile of dusp4 was further shown to correlate precisely with PPARγ during differentiation of 3T3‐L1 and F442A adipocytes. Loss‐of‐function studies are underway to support our working model that dusp4 plays a role in modulating biological outcome of PPARγ activity through covalent modification involving phosphorylation by MAPK.

Grant Funding Source: Support by NIH‐NIDDK (R15‐DK082799)
No effect of the type of sugar on ectopic fat storage (1039.1)
Authors: Lowndes Joshua , Bravo Steven , Sinnett Stephanie , Jordan Richard , Grench Kory , Rippe James
Published: June 2021

Dietary sugar intake has been implicated in the development of insulin resistance and subsequent cardiometabolic disease. It has been proposed the fructose component of commonly consumed sugars may promote the storage of ectopic fat deposits to a greater degree than other types of sugars. The purpose of the present study was to investigate the changes in muscle and liver fat after ten weeks of daily consumption of different types of sugars.

48 weight‐stable (weight change <3% in previous 30 days) individuals aged 20‐60 years old drank sugar‐sweetened low fat milk every day for 10 weeks as part of their usual diet. The amount of milk consumed was individualized for each participant based on the estimated number of calories required to maintain body weight (Miflin St Jeor equation) and random group assignment: Groups 1 and 2 ‐ 9% estimated caloric intake from fructose or glucose respectively added to milk. Groups 3 and 4 ‐ 18% of estimated caloric intake from HFCS or sucrose respectively added to the milk ( 50th percentile for consumption levels of fructose in the USA). Whole body fat percentage was measured by DEXA (GE Lunar) while muscle fat was measured by MRI and liver fat content was measured by MRS (Siemens 3T Skyra).

There was a small increase in weight (166.7 ± 29.4 vs 169.0 ± 31.2lbs, p<0.01) and body fat percentage (32.6 ± 8.9 vs 33.1 ±8.4%, p<0.05) in the entire cohort, but there was no effect of the type of sugar consumed. Small increases were also observed in the vastus lateralis (2.7 ± 0.9 vs 3.1 ±1.2 g/dl, p<0.01) and gluteus maximus (3.6 ±1.1 vs 4.3 ±1.6 g/dl, p<0.001) and in the liver (4.1 ±5.4 vs 4.9 ±6.1 %, p<0.01), but none of these changes were affected by the type of sugar consumed.

These data suggest that short‐term changes in overall body composition are also reflected in the fat content of the liver and muscles, even when the changes are small. Furthermore, in the context of overall fat gain, whether or not the sugar contained fructose had no impact on the magnitude of change.
Addition of protein or fiber to pasta does not alter satiety or mid‐afternoon snacking in healthy men and women (1040.4)
Authors: Korczak Renata , Ahnen Rylee , Timm Derek , Slavin Joanne , Thomas William
Published: June 2021

Protein and fiber can promote satiety and reduce energy intake independently, but few studies have compared the satiating effects of both nutrients in whole foods. This study compared satiety after protein‐enhanced pasta (16g protein, 6g fiber), fiber‐enhanced pasta (11g protein, 8g fiber) or control pasta (11g protein, 6g fiber). In a randomized, double‐blind crossover trial, fasted (4 hours) men (n=18) and women (n=18) ate pastas at 12:00pm along with 500mL of water and rated satiety using Visual Analog Scales at 0, 15, 30, 45, 60, 90, 120 and 180 minutes and palatability at 30 minutes. At 3:00pm, subjects received ad libitum snacks and intake was overtly measured by weight. Breakfast intake was recorded in a food diary and so was intake after men and women left the study visit. Gastrointestinal tolerance was rated by a questionnaire. No differences were found between the pastas for satiety, snacking or gastrointestinal tolerance. Men ate significantly more calories (p=0.007) after the protein pasta versus the fiber pasta (1701 ± 154 vs. 1083 ± 154) with control pasta intermediate (1368 ± 154). The protein pasta was rated less tasty (p=0.03) and less pleasant (p=0.01) than the other two pastas. Increasing the satiating properties of whole foods conveys a challenge as people are influenced by hedonic properties of food.
1, 25‐Dihydroxyvitamin D3/Vit D receptor suppresses brown adipocyte differentiation and mitochondrial biogenesis (1041.6)
Authors: Ricciardi Carolyn , Bae Jiyoung , Hu Pan , Esposito Debora , Komarnytsky Slavko , Chen Jiangang , Zhao Ling
Published: June 2021

Brown adipose tissue, now known to be present in adult humans, is a specialized tissue responsible for heat production through non‐shivering thermogenesis. Understanding of the regulation of the differentiation from the precursors to functional brown adipocytes holds the promise for novel strategies to combat obesity. The purpose of the study was to determine the effects of 1,25‐dihydroxyvitamin D/Vit D receptor on the differentiation and function of brown adipocytes from the immortalized precursor cells derived from interscapular brown adipose tissue of newborn C57BL/6 mice. Here we report that Vitamin D dose‐dependently (1‐100nM) suppressed brown adipocyte differentiation, as measured by both mRNA and protein expression of brown adipocyte markers and oil red O staining of lipid accumulation. Consistently, over‐expression of Vit D receptor also significantly impaired differentiation of the brown adipocytes. Further, results from the cellular bioenergetics measurements showed that suppression of differentiation by Vit D was accompanied by significant impairment of isoproterenol‐induced oxygen consumption rates, maximal respiration and the uncoupled respiration from proton leaks. Together, our results conclude that Vit D signaling suppresses brown adipocyte differentiation and function.

Grant Funding Source: Supported by the University of Tennessee faculty start‐up funds to L.Z.
Fabrication of self‐assembled (‐)‐epigallocatechin gallate (EGCG) ovalbumin‐dextran conjugate nanoparticles and their transport across monolayers of human intestinal epithelial Caco‐2 cells (1044.1)
Authors: Li Zheng , Gu Liwei
Published: June 2021

Nanoparticles have the potential to increase bioavailability of bioactive compounds such as epigallocatechin gallate (EGCG). Ovalbumin was conjugated with dextran using Maillard reaction. The resultant conjugates were self‐assembled with EGCG to form EGCG ovalbumin‐dextran (O‐D) conjugate nanoparticles. Ovalbumin in EGCG O‐D conjugate nanoparticles were further crosslinked by glutaraldehyde. EGCG O‐D conjugate nanoparticles and crosslinked nanoparticles in the aqueous suspension had a particle size of 285 nm and 339 nm, and showed a spherical morphology. The loading efficiency of EGCG in these two nanoparticles was 23.4% and 30.0%, whereas the loading capacity was 19.6% and 20.9%, respectively. EGCG O‐D conjugate nanoparticles maintained the particle size of 183 nm‐349 nm in simulated gastric fluid (SGF) and 188 nm‐291 nm in simulated intestinal fluid (SIF), whereas crosslinked nanoparticles had the particle size of 294 nm‐527 nm in SGF and 206 nm‐300 nm in SIF, respectively. Limited release of EGCG was observed in both nanoparticles systems in simulated gastric and intestinal fluids without and with digestive enzymes. EGCG O‐D conjugate nanoparticles significantly enhanced the apparent permeability coefficient (Papp) of EGCG on Caco‐2 monolayers compared to EGCG solution, suggesting that these nanoparticles may improve the absorption of EGCG.
The effect of individual milk proteins on bioaccessibility of green tea flavan‐3‐ols (1044.12)
Authors: Moser Sydney , Chegeni Mohammad , Liceaga Andrea , Jones Owen , Ferruzzi Mario
Published: June 2021

While information regarding impact of flavan‐3‐ol‐protein interactions on food quality exists, the ability of these interactions to alter bioavailability of health promoting flavan‐3‐ols remains unclear. The objectives of this study were to characterize influence of milk proteins and matrix on in vitro bioaccessibility of flavan‐3‐ols and to assess impact of protein digestion on protein‐flavan‐3‐ol binding. Protein solutions containing sodium‐caseinate (S‐CSN), α‐lactalbumin (α‐LA), β‐lactoglobulin (β‐LG), or non‐fat dry milk (NFDM) were prepared in Jenness Koops (JK) buffer containing milk salts and formulated at 10‐40% (v/v) into green tea beverages containing 0.6 mg/mL total flavan‐3‐ols. Samples were subjected to a three‐stage in vitro digestion to assess flavan‐3‐ol stability and bioaccessibility. Milk protein, most notably S‐CSN, significantly decreased (p<0.05) bioaccessibility of flavan‐3‐ols relative to JK buffer controls (10 relative to 32%). Interestingly, milk minerals significantly increased (p<0.05) flavan‐3‐ol bioaccessibility compared to water formulated controls (32 relative to 18%). Data from in vitro digestions combined with SDS‐PAGE and fluorometric analyses suggest that both milk proteins and minerals may alter flavan‐3‐ol bioaccessibility but normal intestinal digestion likely minimizes the impact of protein interactions.
Compounds from rosemary and Mexican oregano are natural inhibitors of dipeptidyl peptidase‐IV, a target for type‐2 diabetes management (1045.20)
Authors: Bower Allyson , Real‐Hernandez Luis , Berhow Mark , de Mejia Elvira
Published: June 2021

Herbs are a concentrated source of polyphenols. Emerging research shows polyphenols can inhibit the diabetic drug target dipeptidyl peptidase‐IV (DPPiv). The aim was to identify compounds from culinary herbs that may be used in the management of type‐2 diabetes (T2DM). Total polyphenol (TP) and total flavonoid (TF) concentrations as well as DPPiv inhibition were measured and bioassay guided fractionation was used to identify compounds from oregano (OV, Origanum vulgare), marjoram (MO, Origanum majorana), rosemary (RO, Rosmarinus officinalis) and Mexican oregano (LG, Lippia graveolens) that inhibit DPPiv. OV had the highest concentration of TP (430 ± 17 μg gallic acid equiv/mg dry weight, DW) while RO of TF (661 ± 25 μg rutin equiv/mg DW, P<0.01). The herb extracts and their fractions with the lowest IC50 for DPPiv were RO (16 ± 5 μM), LG (31 ± 3 μM), RO fractions (range 0.37‐20.27 μM) and LG fractions (range 1.1‐17.4 μM). LC/MS identified flavonoids in fractions from RO and LG; of these compounds, computational modeling identified hispidulin (‐9.4), eriodictyol (‐8.9) and naringenin (‐8.6) as having the lowest binding affinities (kcal/mol). The IC50 of these compounds were 0.5, 11.2 and 2.8 μM, respectively, and formed H‐bonds to Thr565 and Arg560; hispidulin and naringenin to Lys512, while only hispidulin formed H‐bonds to Asn562. These amino acids may be important sites of DPPiv inhibition by flavonoids. Overall, these data suggest that hispidulin, naringenin and herbs containing these flavonoids are potent inhibitors of DPPiv and should be investigated further in the management of T2DM.

Grant Funding Source: Jonathan Baldwin Turner Fellowship, Division of Nutritional Sciences, University of Il