bluuebunny/crossref_metadata_2025 huggingface dataset:
Confronting Public Nutrition Misconceptions: Using Timelines and Backchannels to Promote Engagement and Critical Reasoning
Authors: Ludy Mary‐Jon , Hamady Carrie , Haar Christine , Plaine Emily
Published: June 2021

Nutrition professionals are inundated with questions about food fads. Students struggle with mechanisms for confronting misconceptions. This innovative educational approach's aim was to promote critical reasoning and elicit reflective inquiry about popular diets using a technology‐rich format. Graduate nutrition students (n=8) participated in a semester‐long, student‐led course using timelines and backchannels. Students used a web app, TikiToki, to create an interactive multimedia timeline profiling food documentaries, customized with embedded images, text, and videos. Students then generated a list of “must‐see” documentaries. Subsequent classes involved viewing documentaries (frontchannel: presentation itself), while simultaneously discussing content on Today's Meet (backchannel: online, synchronous chatroom). Interview data were collected at the end of each class. Qualitative methods were used for analysis. Students generated a timeline profiling 16 documentaries and averaged 187±87 Today's Meet postings per class. They stated this provided a “safe” environment to challenge ideas and reaffirm knowledge of evidence‐based guidelines. Faculty believed multiple learning styles were accommodated by enabling verbally quiet students to express themselves more openly in a virtual classroom. Perceived benefits over Twitter included the private/isolated environment, no account registration, and no need for hashtags. In a society with unrestricted Internet access, timelining and backchanneling serve as meaningful ways to incorporate technology into classes. These techniques present unique methods for disseminating ideas, sharing knowledge, and fostering understanding.
Teachers Don't Know What They Don't Know About Nutrition: A Case for Improving Professional Development
Authors: Linnell Jessica , Smith M. , Brian K. , Scherr R. , Zidenberg‐Cherr S.
Published: June 2021

Self‐efficacy (SE) and lack of nutrition knowledge (NK) are barriers to teaching nutrition. Lesson study is a professional development (PD) model observed to improve SE and NK in other subject areas. The objective of this pilot study was to examine the influence of this PD model in nutrition. Fourth ‐grade teachers (n = 8) participated in lesson study PD on Discovering Healthy Choices, a garden‐enhanced, inquiry‐based curriculum for the Shaping Healthy Choices Program. Pre/post questionnaires were used to evaluate SE and NK, in addition to prior experience teaching nutrition and resources used. Data were analyzed using t‐tests and Spearman's correlations. Changes in SE were not significant, however 3 increased (μ = 10), 4 decreased (μ = ‐9), and 1 did not change. There was a negative correlation between change in SE and those who reported teaching nutrition before (r = ‐0.69). Top resources reported were other teachers and Dairy Council materials, and none reported use of nutrition standards. There were no significant changes in NK; however, 4 increased (μ = 9), and 4 decreased (μ = ‐8). A negative correlation was observed between changes in NK and “not sure” responses (r = ‐0.90). These results generate new questions about PD for teachers, including: Does teaching using a comprehensive nutrition curriculum increase teachers' awareness of the complexity of nutrition? Does teaching nutrition require PD emphasizing basic nutrition and use of evidence‐based resources? These questions should be further examined to develop effective PD for teachers.
The Effect of Almond Consumption on Postprandial Metabolic Response in High‐Risk Pregnant Women
Authors: Lesser Mary , Sawrey‐Kubicek Lisa , Mauldin Kasuen , King Janet
Published: June 2021

The prevalence of metabolic disorders during pregnancy has increased concurrently with the rise in maternal obesity. Obese pregnant women are more prone to chronic metabolic diseases, such as diabetes and cardiovascular disease, following pregnancy. Also, their babies are at greater risk for obesity and metabolic disorders later in life. Almonds provide a healthy source of dietary fat and carbohydrates. A number of clinical trials have also shown that almonds reduce oxidative stress and inflammation, and they promote vascular health and glycemic control. A cross‐over, randomized control trial examining the metabolic response to 2 oz of almond consumption versus dairy fat consumption in two standardized, isocaloric, equal macronutrient composition test meals was done in 18 overweight/obese (pre‐pregnancy BMI = 25‐40) women during 30‐36 weeks of gestation. Circulating concentrations of glucose, triglycerides, insulin, glucagon, and GLP‐1 were measured in the fasting state and during a 5‐hour postprandial period. Hormones involved in appetite, glucose and lipid metabolism (leptin, ghrelin, and c‐peptide) were also measured. Replacing cream cheese with almonds in an isocaloric, equivalent fat and carbohydrate meal had a minimal effect on postprandial lipids, glucose, and hormones. This suggests that the positive metabolic effects of almonds in previous feeding studies were likely related to shifts in the energy and macronutrient content of the overall diet in association with consuming almonds as a fiber‐rich, fat source. (Supported by the Almond Board of California).
Inflammatory markers and Nutrients intake in Korean patients with non alcholic fatty liver
Authors: Kim Soon , Lim Hee , Kwon Yeon
Published: June 2021

The purpose of this study was to examine the correlations between fatty liver disease, obesity, blood inflammatory markers, biochemical indicators, and diet. The subjects included 40 males and 40 females who visited the Health Promotion Center at the S University Hospital. Among the participants, 22 males (55.0%) and 10 females (25.0%) were diagnosed with non‐alcoholic fatty liver disease. The study reported that subjects in the fatty liver group (FLG) had a significantly higher mean body mass index, body fat content, and WHR than those in the non‐fatty liver group (NFLG). In addition, subjects in the FLG had higher levels of serum ALT, γ‐glutamyl transpeptidase, cholesterol, and triglycerides than those in the NFLG. The mean level of adiponectin, an inflammatory marker, was significantly lower in male and female subjects in the FLG than those in the NFLG, and the C‐reactive protein and tumor necrosis factor‐α levels were significantly higher in female subjects in the FLG than those in the NFLG. The subjects in the FLG had significantly higher levels of metabolic syndrome indicator, and approximately 60% of the female subjects in the FLG also had metabolic syndrome. Male subjects in the FLG had significantly higher intake of sodium, vitamin A, and saturated fatty acids but lower intake of iron than those in the NFLG. Female subjects in the FLG had significantly higher intake of calories, carbohydrates, iron, sodium, cholesterol, and saturated fatty acid but lower intake of vitamin B6 than those in the NFLG.The adiponectin level was significantly lower in all subjects in the FLG than those in the NFLG.
Changes in Microbiome Profiles and Nutrient Absorption in Intestine by Soy Protein
Authors: Lim Jinkyu , Kim Sunghee , Choi Jaesun , Chen Xiaocen
Published: June 2021

Soybean has good nutritious values with high content of proteins cultivated all over the world. Form our previous studies, we found that soy protein is resistant to in vitro digestion by proteases from digestive tract, bacteria and fungi. We call the protease resistant soy protein or protease resistant food protein (PRFP) and assume that the PRFPs interact with the intestinal tracts, nutrients, or microbes in the intestine to influence the physiological status of the body. We tested the interaction between PRFP and microbes, and nutrients, such as cholesterol and triglyceride. The interference of the fluorescence labeled bacteria‐intestinal cell interaction was shown in the presence of soy proteins. Plus, we observed the structural change in the microvilli of small intestine of mice led soy diet. For the preliminary studies, we also analyzed the microbiome profiles in the fees of the mice fed soy proteins using metagenomic sequencing. This study suggests that soy protein is a good source of nutrient but also a biological polymer to influence physiological change and microbiome changes.
Functional Studies of the Putative Organic Cation Transporter SLC22A23
Authors: Chaity Nazia
Published: June 2021

Introduction

SLC22A23 (Solute Carrier 22 family 23) is an orphan gene that encodes for a putative organic cation membrane transporter protein. The gene has been repeatedly associated to inflammatory bowel disease however; the gene structure in human and its function is still uncharacterized. Inflammatory bowel disease (IBD) is a chronic disease that is steadily increasing worldwide with the highest prevalence in Canada.

Objectives

The proposed research aims to determine the tissue expression and characterize the subcellular location of the SLC22A23 protein, in mammalian cells.

Methods

Human SLC22A23 has been subcloned by Gateway® Technology and tagged with green fluorescent protein (GFP) and transfected in three cell lines‐ CHO, MDCK and Caco‐2. Membrane localization of SLC22A23 has been performed using confocal fluorescence microscopy. And to determine the tissue expression Real‐Time polymerase chain reaction has been used.

Results

Human SLC22A23 protein is located in the cell membrane and is abundantly expressed in various tissues in human body with high expression in brain, kidney and intestine.

Conclusion

The characterization of the SLC22A23 gene will provide the basis to determine which genetic variation is causative in the development of IBD in future studies. The correct sub‐cellular location will confirm that the human SLC22A23 is a membrane transporter protein (as known from the rat orthologous). Further functional studies of this gene will explicate the mechanism in the development of IBD and will contribute to the understanding which metabolite/nutrient imbalance plays a role in the development of IBD.
Placental Transcriptomic Changes Due to Maternal Exercise are Dependent on Maternal Diet and Offspring Sex
Authors: Shankar Kartik , Borengasser Sarah , Faske Jennifer , Kang Ping , Zhong Ying , Thakali Keshari
Published: June 2021

While exercise (EX) is beneficial during pregnancy for both mother and child, little is known about the mechanisms by which maternal (MAT) EX mediates changes in utero. Here we examined transcriptomic signatures in the placenta following MAT EX. Female C57BL/6 mice were fed either a control or HFD (45% fat) starting at 6 wk of age, with (EX) or without access to voluntary running wheels. Mice had 24‐h access to wheels for 10 wk prior to conception until the end of the study (18.5 days post coitum). Placenta were collected from individual pups for microarray analysis in a sex‐identified fashion. In control‐fed mice, EX affected expression (± 1.4‐fold, p < 0.05) of 661 transcripts in female placenta (vs. 283 transcripts in males), while in female placenta from HFD‐fed mice,1.7‐fold more genes had altered expression due to MAT EX. Gene‐set enrichment analysis of male placenta from control‐fed dams showed that MAT EX led to enrichment of genes involved in neurotransmitter levels and glutamate signaling and decreases in lipid biosynthesis and transport, while EX in female placenta from the same dams led to increases in genes involved in muscle and brain development and decreases in hormone metabolic processes. On HFD, female placenta had a prominent decrease in oxidative phosphorylation related genes, and both sexes showed decreased glutathione metabolism genes. Specific genes were confirmed using qPCR. Our results suggest that effects of MAT EX on the placenta and presumably on the offspring are influenced by maternal habitus and are sexually dimorphic. (NIH‐NIDDK R01‐084225 and USDA‐ARS CRIS 6251‐51000‐007‐04S).
Fatty Acid Patterns and the Risk of Prostate Cancer in the Physicians' Health Study
Authors: Yang Meng , Ayuningtyas Azalea , Kenfield Stacey , Sesso Howard , Campos Hannia , Ma Jing , Stampfer Meir , Chavarro Jorge
Published: June 2021

Analyzing blood fatty acid (FA) patterns could better capture FA interactions and aid in characterizing the relation to prostate cancer (PCa). We aimed to assess data‐derived FA patterns and PCa risk. We conducted a nested case‐control among men in the Physicians' Health Study. FA levels were measured in pre‐diagnostic whole blood samples of 476 cases and their matched controls to identify 28 individual FA. FA patterns were identified using principal component analysis. Conditional logistic regression was used to estimate RRs and 95% CIs of PCa across quintiles of FA patterns. Two patterns explaining 40.9% of total variation were identified. Pattern 1 was characterized by high levels of trans‐FA and α‐linolenic acid along with low levels of n‐3 PUFA. Pattern 2 was characterized by high levels of 14 and 16 carbon saturated FA and MUFA along with low levels of α‐linolenic acid and saturated FA of 18 or more carbons; this pattern largely reflects de novo lipogenesis. There was a positive but nonsignificant association between pattern 1 and PCa risk (RRQ5 vs Q1=1.37, 95%CI: 0.91‐2.05). Pattern 2 was statistically significantly associated with higher PCa risk (RRQ5 vs Q1=1.63, 95%CI: 1.04‐2.55). This association was similar across tumor stage, grade, and clinical aggressiveness categories. The two FA patterns were consistent with known interactions between FA intake and metabolism. A pattern suggestive of higher activity in the de novo lipogenesis pathway was related to higher PCa risk.
Metabolic Alterations in Mice Lacking a Major Folate Catabolic Enzyme
Authors: Fekry Baharan , Krupenko Natalia , Krupenko Sergey
Published: June 2021

Aldh1l1 (10‐formyltetrahydrofolate dehydrogenase, FDH) is a major folate enzyme, regulating the overall flow of one‐carbon groups towards biosynthetic processes. We have generated mice with the targeted knockout of Aldh1l1 and characterized their metabolic and protein expression profiles using the metabolomics approach and NanoString technology. We have observed that KO mice have significantly elevated 10‐formyltetrahydrofolate and depleted tetrahydrofolate pools in the liver, in agreement with Aldh1l1 catalytic function. The total liver folate was depleted in KO mice, which could indicate the lack of folate protection function proposed for Aldh1l1. In agreement with this decrease, levels of formiminogluatmate were elevated in KO mice implying the functional folate deficiency. Interestingly, the total folate in blood (represented mostly by 5‐methyltetrahydrofolate) was not changed. The metabolomics approach revealed changes in serine and glycine metabolism in these mice, as well as broad alterations in lipid metabolites. In agreement with the proposed function of Aldh1l1 as a candidate tumor suppressor, significant alterations in tumorigenesis‐related genes and pathways were observed including the elevation of mRNA levels of TGFb family members, certain transcription factors, and enzymes implicated in cancer cell metabolism (most noticeably glucose‐6‐phosphate dehydrogenase and phosphofructokinase). Furthermore, decreased protein levels of p53, JNK2 and CerS6, previously reported as mediators of Aldh1l1 metabolic effects, were seen in livers of Aldh1l1 KO mice. These findings underscore Aldh1l1 as a crucial regulator of metabolic pathways in mammalian cells. This work was supported by NIH grant DK54388.
A Lycium Chinense Beverage Alleviated Inflammation in Overweight Subjects with High LDL Cholesterol Level: a Double‐Blinded, Randomized, Placebo‐Controlled Study
Authors: Ahn Youngsook , Lee You jin , Lim Yeni , Kwon Oran
Published: June 2021

Obesity and high level of low density lipoprotein cholesterol (LDL‐C) are considered as major risk factors of the metabolic syndrome and cardiovascular disease. In this study, we hypothesized that an administration of a Lycium chinense (LC) beverage may exert anti‐inflammatory effect in overweight subjects with high LDL‐C level. 53 eligible subjects were randomized to either a LC (n=26) or a placebo (n=27) for 8 weeks in a double‐blinded trial. LC was standardized with anthocyanin. Transcriptomics analysis of inflammatory mediators and signaling molecules, lipid metabolism, blood cell differentiation, plague formation, coagulation, and antioxidant makers were applied using quantitative real‐time reverse transcription‐polymerase chain reaction array. Postprandial plasma level of inflammatory cytokines were assessed by proteomics using Luminex assay. Subjects who consumed a LC for 8 weeks showed a significant down‐regulation of mRNA levels related to oxidation and inflammation compared to the placebo. Consistent results were found in proteomics, showing significant increases of the postprandial plasma levels of tumor necrosis factor‐α and fms‐related tyrosine kinase 3 ligand in the LC compared with the placebo. In addition, 11 subjects were received the postprandial test after co‐administration of a bolus of fat challenge formula and a LC at the final visit. Postprandial blood samples were obtained at 0, 2, 4, 6, and 8 hours after administration and the results will be presented as well. Taken together, these results suggest that daily consumption of a LC beverage may provide a potential health benefits of reducing inflammatory cytokines in subjects overweight and high level of LDL‐C.
Antioxidant Characteristics of Algal Sagassum muticum Yendo Fensholt And Ishige okamurae Yendo Based on Its Total Phenolic Content
Authors: Lee Je‐Hyuk
Published: June 2021

Sagassum muticum Yendo Fensholt and Ishige okamurae Yendo are marine algae used as a material for local food in Jeju‐island, South Korea. Antioxidant characteristics of Sagassum muticum Yendo Fensholt and Ishige okamurae Yendo based on its total phenolic content was investigated in this study. Scavenging activity for DPPH radical (100 μM) in Sagassum muticum Yendo Fensholt and Ishige okamurae Yendo extract (300μg/ml) was approximately 69.9% (174.06 μg/ml=IC50) and 89.3% (29.9 μg/ml=IC50), respectively. As considering total phenolic content (approximately 37.98±0.53 and 120.80±6.05 mg GAE/g dw, respectively) in extract of Sagassum muticum and Ishige okamurae, total phenolic compound (11.39 and 36.24 μg/ml) in Sagassum muticum and Ishige okamurae had approximately 69.9% (IC50=4.184 μg/ml, more than 41 times) and 89.3% (IC50=3.094 μg/ml, more than 11 times) for DPPH radicals. Reducing power of Sagassum muticum and Ishige okamurae extract (300μg/ml) was approximately 0.102±0.0001 and 0.126±0.00007 (OD700), and a slope with 1.591E‐4 and 3.169E‐4, respectively. However, reducing power of total phenolic compounds (11.39 and 36.24μg/ml) in Sagassum muticum and Ishige okamurae had approximately 0.102±0.0001(OD700) and 0.126±0.00007(OD700) showing linearity with a slope of 4.187E‐3 and 2.623E‐3, respectively. ABTS and ORAC in Sagassum muticum extract was 00.56±12.77 and 181.48±8.32 μmol TE/g dw, whereas their total phenolic compounds had approximately 7913±334 and 4778±219 μmol TE/g dw, respectively. Additionally, Ishige okamurae extract showed 748.27±11.13 and 328.82±13.08 μmol TE/g dw of ABTS and ORAC, whereas their total phenolic compounds had approximately 6194±92.1 and 2722±108.2 μmol TE/g dw, respectively.
Effect of oral administration of DHA on the remodeling of calcium handling proteins in congestive heart failure in male rats
Authors: Yamanushi Tomoko , Kabuto Hideaki , Akiyama Kaoru , Tsushima Tadahiro , Misawa Yoshihisa
Published: June 2021

Calcium (Ca2+) cycling is essential in cardiac contractile function, and remodeling of calcium handling is thought to be one of the factors contributing to the dysfunction in heart failure (HF). This study investigated the effect of DHA administration on remodeling of Ca2+handling proteins in congestive heart failure (CHF). Male rats were divided into 4 groups (n=8 for each group): C (normal, administered saline); D (normal, administered DHA‐ethyl ester (DHA‐Et)); H (CHF, administered saline); HD (CHF, administered DHA‐Et). CHF was produced by injection of monocrotaline (MCT) into 5‐week‐old rats. Starting from 3 days before MCT injection, saline or DHA‐Et wasorally administered(1g/kg body weight) daily. The right ventriclewas removed on day 24, and quantitative PCR was used to measure the expression of transcripts.The expression of sarcoplasmic reticulum Ca2+‐ATPase (SERCA) 2a, which is responsible to transport back Ca2+ to sarcoplasmic reticulum (SR), was significantly decreased in group H compared to groups C, D and HD (P 蠄 0.05). The expression of ryanodine receptor (RYR) 3, which releases Ca2+ from SR, in group H was significantly increased compared to groups C and D (P 蠄 0.05), and also tended to be increased in group H compared to group HD (P=0.06).The expression of Cav1.2, a component of L‐type Ca2+ channel,was significantly decreased in group H compared to groups C and D (P 蠄 0.05). Remodeling of intracellular Ca2+ handling may be caused by these changes in gene expression in CHF. Administration of DHA‐Et may improve abnormal Ca2+ handling through recovery of the expression of these proteins.
Wild Blueberry Consumption (V. angustifolium) Attenuates Local Inflammation in the Perivascular Adipose Tissue (PVAT) of the Obese Zucker Rat (OZR)
Authors: Klimis‐Zacas Dorothy , Vendrame Stefano , Kristo Aleksandra
Published: June 2021

PVAT has emerged as critical in vasoregulation and a link between systemic and local vascular inflammation.

The role of wild blueberries (WB) was examined on Phenylephrine‐induced vasoconstriction (10‐8 to 3x10‐6M) in aortic rings with or without PVAT in 12 week‐old OZRs (n=16) and their lean littermates (LZR) (n=16) on a control (C) or an 8% WB diet for eight weeks. Interleukin (IL)‐8, monocyte chemoactractant protein (MCP)‐1, tumor necrosis factor (TNF)‐a and adiponectin concentrations were measured in PVAT by ELISA.

Maximal vasoconstriction force (Fmax) in aorta without PVAT was significantly lower in OZR‐C compared to LZR‐C (0.41 ± 0.05 and 0.71 ± 0.06 g, respectively). Following WB diet, a significant increase in Fmax was observed in the OZR group (0.54 ± 0.06 g). In aortic rings with PVAT, Fmax was significantly lower in all groups and all treatments (0.31 ± 0.06 OZR‐C, 0.30 ± 0.05 OZR‐WB, 0.29 ± 0.03 LZR‐C and 0.30 ± 0.04 g LZR‐WB), but no difference was observed between treatments.

In PVAT, concentrations of MCP‐1, TNF‐a and adiponectin were significantly higher in OZR group compared to LZR (+102%, +108%, and +45%, respectively). Following the WB diet, concentrations of IL‐8 in PVAT were significantly lower in both OZR (–37%) and LZR (–30%), while adiponectin concentrations significantly increased in both OZR (+11%) and LZR (+16%). MCP‐1 concentrations significantly decreased (–31%) in the PVAT of OZR with WB diet.

Thus wild blueberries have the potential to attenuate local inflammation in PVAT with possible implications on arterial function.

Funded by the Maine Wild Blueberry Commission
Effect of Rosemary (Rosmarinus Officinalis L.) Supplementation on Cardiac Remodeling after Myocardial Infarction in Rats
Authors: Murino Rafacho Bruna , dos Santos Priscila , Gonçalves Andrea , Fernandes Ana Angelica , Gaiolla Paula , Zornoff Leonardo , Minicucci Marcos , de Paiva Sergio
Published: June 2021

Aim

To analyze the influence of rosemary (R) consumption on cardiac remodeling after myocardial infarction (MI). Material: Male Wistar rats were allocated into: 1) Sham group fed standard chow (SA0, n=6); 2) sham group fed standard chow + 0.02% of dried R (SA002, n=6); 3) sham group fed standard chow + 0.2% of dried R (SA02, n=6); 4) MI rats fed standard chow (IA0, n = 6); 5) MI rats fed standard chow + 0.02% of R (IA002, n=6); 6) MI rats fed standard chow + 0.2% of R (IA02, n=6). After 3 months, heart tissue was collected and evaluated for collagen and myocite cross seccional area (CSA), lipid hydroperoxide concentration (LH) and enzyme activity of gluthatione peroxidase (GPx), superoxide dismutase (SOD), citrato sintase (CS), lactate dehydrogenase (LDH), pyruvate dehydrogenase (PDH), β‐Acyl CoA dehydrogenase (β‐OH‐acil CoA‐DH), ATP sinthase, Complex I and II of respiratory chain. Comparisons were made by two‐way ANOVA. Values were presented as mean ± standard error. Results: MI rats showed higher CSA and collagen content, higher cardiac LH and SOD, and lower GSH‐Px and Complex I activity than Sham rats, suggesting hypertrophy, oxidative stress and mitochondrial dysfunction 3 months after MI. Rosemary supplementation with 0.02% reduced CSA. Both doses of R reduced LH, SOD and LDH, increased GSH‐Px, CS, β‐OH‐acil CoA‐DH and Complex I activity after MI. Treatment with 0.2% of R increased Complex II.

Conclusion

Chow supplementation with rosemary attenuated cardiac remodeling after myocardial infarction in rats. Financial support: FAPESP.
Anthocyanin‐rich Black Elderberry Extract Improves Markers of HDL Function and Reduces Aortic Cholesterol in Hyperlipidemic Mice
Authors: Farrell Nicholas , Norris Gregory , Lee Sang Gil , Chun Ock , Blesso Christopher
Published: June 2021

Atherosclerosis is a chronic inflammatory disease and is associated with impairments in high‐density lipoprotein (HDL) function, including reductions in HDL‐cholesterol, antioxidant and anti‐inflammatory activities. Anthocyanins are polyphenols with anti‐inflammatory properties. We investigated whether an anthocyanin‐rich black elderberry extract (Sambucus nigra) (BEE) (13% anthocyanins) would protect against inflammation‐related impairments in HDL function and atherosclerosis in apoE‐/‐ mice. We fed an AIN‐93M diet supplemented with 1.25% (wt/wt) BEE or control diet to 10 week old male apoE‐/‐ mice for 6 weeks. The BEE fed to mice was rich in cyanidin 3‐sambubioside (~9.8% wt/wt) and cyanidin 3‐glucoside (~3.8% wt/wt). After 6 weeks, serum lipids did not differ significantly between groups, while aspartate transaminase (AST) and fasting glucose were reduced in BEE‐fed mice. Hepatic and intestinal mRNA changes with BEE‐feeding were consistent with an improvement in HDL function (Apoa1, Pon1, Saa1, Lcat, Clu) and a reduction in hepatic cholesterol levels (increased Ldlr and Hmgcr, reduced Cyp7a1). In BEE‐fed mice, serum paraoxonase‐1 activity was significantly higher. In addition, mice fed BEE had significantly lower serum chemokine (C‐C motif) ligand 2 (CCL2) and tended to have lower serum thiobarbituric acid reactive substances compared to control‐fed mice. Notably, we observed significant reductions in total cholesterol content of the aorta of BEE‐fed mice, indicating less atherosclerosis progression. These findings suggest that black elderberry may be useful to prevent HDL dysfunction associated with chronic inflammation. [Supported by the USDA Hatch Project #1003343].
Peptides from Common Bean (Phaseolus vulgaris L.) non‐Digestible Fraction Inhibit Angiotensin‐I Converting Enzyme by Interacting with Amino Acids in its Catalytic Cavity
Authors: Luna Vital Diego , Gonzalez de Mejia Elvira , Mendoza Sandra , Loarca‐Pina Guadalupe
Published: June 2021

Five major peptides (GLTSK, LGSNK, GEGSGA, MPACGSS and MTEEY) from non‐digestible fraction (NDF) of common bean have demonstrated antiproliferative activity against HCT116 and RKO human colorectal cancer cells. The aim was to evaluate angiotensin‐I converting enzyme (ACE) inhibitory activity and antioxidant capacity of peptides in common bean. For ACE inhibition, hippuril‐histidyl‐leucine was used as substrate and the product (hippuric acid) was quantified by HPLC. Antioxidant capacity was evaluated through FRAP, ABTS and DPPH assays. The inhibition of ACE ranged from IC50=33 to 91 µg of peptide/mL. Peptides inhibited ACE activity by interacting with residues His353, Ala354, Glu384, Glu411 and Tyr523 inside the catalytic cavity of the enzyme as determined by molecular docking. The peptide MPACGSS had the highest (p<0.05) antioxidant activity: 421.6 µmol FeSO4/mg (FRAP), 561.4 μmol Trolox/mg (ABTS) and 748.4 μmol Trolox/mg (DPPH). Since ACE inhibition and antioxidant activity may be involved in colorectal chemopreventive mechanisms, the results support the notion that peptides in common bean can contribute to colon health.

Supported by University of Illinois and University of Queretaro research program
Formation of Urolithins in Normal Volunteers Following Pomegranate Extract Consumption for 28 Days is Associated with Differences in the Gut Microflora Population
Authors: Henning Susanne , yang jieping , Carlson Paula , Finegold Sydney , Heber David , Li Zhaoping
Published: June 2021

Although pomegranate ellagitannins are highly bioactive in vitro, they are not absorbed intact but are hydrolyzed in the intestine to ellagic acid and urolithins. There are known interindividual differences in the intestinal formation and urinary excretion of urolithin A (UA). To investigate the difference in gut microbiota between UA producers and non‐producers 20 healthy volunteers consumed pomegranate extract for 4 weeks. Urine and stool samples were collected at baseline and 4 weeks for urolithin analysis by HPLC and DNA sequencing with Illumina MiSeq analysis. The following differences (percent of total bacteria) in baseline gut microflora phyla were seen between UA producers (n=14) and non‐producers (n=6): Verrucomicrobia (genus Akkermansia) (0.2% vs.6.6%); Actinobacteria (5.2 %vs.1.8%); Firmicutes (68% vs.57%); Bacteroidetes (23%vs.26%) and the Firm/Bact ratio (2.96 vs. 2.19). The abundance of Proteobacteria was significantly increased (3.1% vs. 5.5%) and Actinobacteria decreased (1.8% vs 1.4%) comparing baseline to 4 weeks in producers. The production of urolithins was positively associated with the abundance of Akkermansia, a bacteria previously shown to be inversely correlated with body weight in rodents and humans. Probiotic bifidobacteria were generally not affected by pomegranate extract. Pomegranate extract consumption over 28 days induced changes in the microbiome to enhance production of urolithins but the profile of microflora was stable and different between producers and non‐producers. Future clinical studies of pomegranate juice and extract may benefit from stratifying between producers and non‐producers.
Granzyme B Mediates Both Direct and Indirect Cleavage of Extracellular Matrix in Skin After Chronic Low‐Dose Ultraviolet Light Irradiation
Authors: Parkinson Leigh , Toro Ana , Zhao Hongyan , Granville David
Published: June 2021

Extracellular matrix (ECM) degradation is a hallmark of tissue aging, as well as many other chronic inflammatory diseases that can lead to a loss of function. Granzyme B (GzmB), a serine protease that is expressed by a variety cells, has been shown to accumulate in the extracellular space during chronic inflammation and cleave a number of proteins. Using a chronic low‐grade UV‐irradiation murine model, we hypothesized that GzmB contributes to skin damage and ECM degradation through processes involving both direct ECM cleavage and indirect ECM cleavage through the induction of other proteinases. Wild‐type and GzmB‐knockout (KO) mice were repeatedly exposed to minimal erythemal doses of solar simulated UV‐irradiation for up to 20 weeks. GzmB expression was significantly increased in wild‐type UV‐treated skin compared to non‐irradiated controls. GzmB deficiency significantly protected against the formation of wrinkles and the loss of dermal collagen density. GzmB‐KO mice were also protective against the loss of fibronectin (FN) in vivo, and FN fragments were released from the cell‐derived matrix of cultured fibroblasts after GzmB treatment. GzmB‐mediated FN fragments increased the release of metalloproteinase (MMP)‐1 and MMP‐3 from fibroblasts. Furthermore, GzmB cleavage of decorin predisposed collagen fibrils more susceptible to attack by MMP‐1. Finally, GzmB reduced the barrier function of a monolayer of keratinocytes in vitro as measured by electrical impedance. Collectively, these findings indicate a significant role for GzmB in ECM degradation, which may have implications in many aged‐related dermatological indications.
β1‐Integrin, KCa3.1 and TRPC4 Channels Play Complementary Roles in Alveolar Epithelial Repair
Authors: Chebli Jasmine , Girault Alban , Prive Anik , Trinh Nguyen Thu Ngan , Maille Emilie , Grygorczyk Ryszard , Brochiero Emmanuelle
Published: June 2021

Extensive alveolar epithelial injury and remodelling are typical features of acute respiratory distress syndrome (ARDS). It has been established that epithelial regeneration, and secondary lung oedema resorption, are crucial for the resolution of ARDS parameters. Interestingly, many evidences indicate that K+ channels are regulating epithelial repair processes; however the role of Ca2+‐activated KCa3.1 channels in alveolar repair has never been investigated before.

Wound healing assays first demonstrated that the repair rates of primary rat alveolar cell monolayers were increased on a fibronectin matrix compared to non‐coated supports. KCa3.1 inhibition impaired the fibronectin‐stimulated wound‐healing rates, as well as cell migration and proliferation, but was without effect in absence of coating. We then evaluated a putative relationship between KCa3.1 and the β1‐integrin, which is activated by fibronectin. Immunofluorescence and co‐immunoprecipitation assays indicated a cellular co‐distribution and a physical link between the two proteins. In addition, KCa3.1 and β1‐integrin membrane expressions were both increased on a fibronectin matrix. We also showed increased [Ca2+]i as well as enhanced expression of TRPC4, a non‐voltage gated Ca2+ channel of the large TRP channel family, in the presence of fibronectin. Finally, wound healing assays showed that TRPC4 function is also involved in alveolar repair.

Altogether, our data demonstrate for the first time a complementary role of β1‐integrin, KCa3.1 and TRPC4 channels in the regulation of alveolar repair processes.

Funded by CIHR, FRQS and NSERC.
ASSESSMENT OF THE RELATIONSHIP BETWEEN LEARNING PREFERENCES, INNOVATIVE TEACHING TOOLS, AND ACADEMIC PERFORMANCE IN A MEDICAL PHARMACOLOGY COURSE IN COLOMBIA.
Authors: Mantilla Rivas Jose , Daza‐Zuniga Camilo , Rodriguez‐Pulido Astrid , Pena Silva Ricardo
Published: June 2021

The impact of learning preferences on academic performance and use of innovative tools to innovate education, among medical students, is not clear. In our medical pharmacology course we have introduced several elements to foster student learning. Particularly, some of our lectures are using a classroom management system: ¨learning catalytics¨ that increases engagement of students during lectures.. We have also introduced crossword puzzles to help students to prepare for tests. Fifty‐three second‐year medical students taking the Pharmacology course in Universidad de Los Andes were interviewed according to the Gainen Learning Preferences Questionnaire. Their responses were then plotted against their average grades in the course and against the weighted average for the exams, and a linear regression was applied. The average grade mean was 3,71/5 (SD=0,437) and the mean value for the Gainen Questionnaire was 3,39/5 (SD=0,367). The point estimator for the linear regression was 0,1643 (R2=0,01899), suggesting there was no relationship between academic performance and learning preferences. Students scored highly the value of novel educational tools (crossword puzzles and learning catalytics). No relationship however was found between academic performance or learning preferences, and the value of novel educational tools. Further studies are needed in order to establish the relationship between learning preferences and academic performance in pharmacology courses. Importantly, most students in our pharmacology course appear to value and benefit from the use of innovative teaching and study methods.
Evaluating the Effectiveness of Pretest and Posttest Model of Active Learning in a Medical School
Authors: Mehnaatamai Mohanram Arun , Zhong Qing , Singh Tarun , Jagadeesh Ashraya
Published: June 2021

Effective medical education is extremely challenging in these times of high competitiveness. With the evolving curriculum changes and shorter contact hours, it is becoming extremely difficult to perform TBL sessions and Flipped Classroom sessions of active learning. In order to strike a balance between short contact hours and effective learning, we tried adding a Pretest and a Posttest to the traditional didactic lecture session. This method works on the principle that by including a multiple‐choice questions based Pretest and Posttest session before and after the didactic lecture respectively, the educator can heighten the attentiveness of the students and make them actively seek information during the lecture session. In order to test this hypothesis, we divided the pharmacology class into two groups namely Group A and Group B and conducted a cross‐over study. On Day 1, Group A was given a Pretest and a Posttest while Group B was given only a Posttest. On Day 2, Group A was given only a Posttest while Group B was given a Pretest and a Posttest. The tests were scored, totalled and compared. Analysis of the results showed that the groups that took both a Pretest and a Posttest significantly scored higher percentages and passing grades compared to the groups that took only the Posttest, albeit not statistically significant. This study clearly demonstrates the advantage of sensitizing the student's minds with a Pretest before the lecture begins. Based on the results of this study, by including a Pretest and a Posttest session to the routine didactic lecture, we can help students not only gain considerably from the lectures but also measure the same.
Natural Product Biosynthesis ‐ A Treasure Trove of Multitalented Flavoenzymes
Authors: Moore Bradley
Published: June 2021

Biosynthetic enzymology is alive and well thanks to the bioinformatics explosion associated with the specialized metabolism of natural product chemicals. Nowhere is that more evident than with flavin‐dependent enzymes, which catalyze a remarkable diversity of biochemical reactions that only appear to be growing in number and wonderment as more natural product biosynthetic pathways are discovered and characterized. The ever versatile flavin cofactor supports a myriad of redox reactions that go well beyond simple hydroxylation and dehydrogenation reactions to support diverse oxidative reactions such as C–H activations, Baeyer‐Villiger ring expansions, oxidative Favorskii rearrangements, and perhalogenation reactions. In the case of the enterocin “favorskiiase”, new flavin biochemistry involving a previously unrecognized superoxidized flavin‐N5‐oxide cofactor is employed as the oxygenating species that may be more widespread in nature. Examples from the author's laboratory will highlight the growing appreciation for flavin as biology's most versatile cofactor.
Effects of Adolescent Nicotine Exposure on Adult Cocaine Reward, Aversion and Self‐Administration
Authors: Pomfrey Rebecca , Bostwick Tamaara , Wetzell B , Riley Anthony
Published: June 2021

These studies examined adolescent nicotine preexposure effects on the rewarding and aversive effects of adult cocaine use and the relationship of these changes to self‐administration. In Experiment 1, rats on postnatal day (PND) 28 were divided into two groups and given daily 0.6 mg/kg injections of nicotine or vehicle until PND 42. Rats were given locomotor tests to examine nicotine‐induced sensitization. They were allowed to age untreated to PND 76, implanted with jugular catheters and allowed to recover for at least 5 days. On PND 90, cocaine self‐administration (0.25 or 0.75 mg/kg), progressive ratio (PR) responding, extinction and cue‐induced reinstatement testing began. Experiment 2 explored nicotine's impact on the balance of cocaine's affective properties. Rats (PND 28‐ 75) were treated as described above. On PND 76, they were tested for the aversive and rewarding effects of cocaine in a combined conditioned taste avoidance (CTA)/place preference (CPP) procedure. Rats were given access to a novel saccharin solution, injected with cocaine (5.6, 10 or 18 mg/kg) then placed in CPP chambers. In Experiment 1, the nicotine group showed sensitized locomotor activity over trials compared to controls, showing that the nicotine was behaviorally active. All rats in Experiment 1 showed clear, dose dependent responding during acquisition, PR, extinction and reinstatement, with no nicotine preexposure effect. In Experiment 2, rats showed a significant CTA trial x Drug effect (p = .012) with cocaine groups consuming less saccharin over trials with no preexposure or dose group differences. There was no significant CPP or interaction with preexposure. These studies suggest that adolescent nicotine preexposure does not have an impact on adult cocaine self‐administration or its affective properties under these parametric conditions.
Role of NKCC2A in Nervous system Activity
Authors: annu Kavya , Lucot James , Garrett Teresa , DiFulvio Mauricio , Castrop Hayo
Published: June 2021

The NKCC2A channel is a splice variant of the Na+K+2Cl‐ co‐transporter 2 (Slc12a1), which is abundantly expressed in macula densa and apical membrane of the tubular cells in the kidney but has not been reported in nervous tissue. This study was designed to determine if the NKCC2A absence altered behavior and nervous system activity in mice. NKCC2A wild type (WT), heterozygous (HE) and homozygous (HO) mice were tested in a behavioral battery in which there was a significant decrease in immobility in the Forced Swim Test (FST), a measure of depression‐like behavior. There were no differences in open field, Y‐maze, plus maze, pre‐pulse inhibition, functional observational battery or the rotarod test. Brain regions and adrenals were homogenized and catecholamines were measured using HPLC coupled to electrochemical detector. The serotonin metabolite to parent ratio in HE was elevated in rostral brainstem, significantly lower in the HO in the caudate nucleus with a non‐significant trend toward the same pattern in the amygdala. There was an increase in the intraneural but not the extraneural metabolite to parent ratio for dopamine. There were significantly lower levels of epinephrine and norepinephrine in the adrenals of HO, with the HE having intermediate levels. The pattern of results is consistent with the decreased immobility in the FST suggesting effects in brain areas involved in depressive‐like behavior. Further experiments will examine the biogenic amine use in nucleus accumbens, frontal cortex, hypothalamus, brain stem and urine. We will also employ a different test for depressive‐like behavior that has different false positives. These results implicate NKCC2A in brain and adrenal function in addition to its known role in the kidney.
CXCR4 Gradient Sensing Generates Unique Signaling Kinetic Signatures of Akt and p53 Pathways, and Drives ERK to Invadopodia in Metastatic Breast Cancer Cells
Authors: Winant Bryce , Freed Jacqueline , Stewart Joshua , Sobel Michael , Campbell Mallory , Jancina Anastasia , Myers Kenneth , Moore Catherine
Published: June 2021

During development, the chemokine receptor CXCR4 is essential for migration of neuronal and hematopoietic cells upon SDF gradient sensing. During disease, the SDF‐CXCR4 axis is critical for metastasis by promoting tumor cell invasion into distal SDF‐expressing organs. Despite this, the pathways downstream of motility inducing SDF gradients are unknown. Previously, we identified unique pathways associated with SDF gradient sensing, including a novel Arf6‐ERK motility pathway and a sustained MAPK pathway atypical for GPCRs but reminiscent of GFR signaling during malignancy. Here we extend these findings using microarray and IB, and identified that gradient SDF also leads to sustained Akt activation, and both the Akt and ERK pathways are inhibited by wortmannin or LY294002. Notably, both pathways regulate processes critical to malignancy including invadopodia formation. Additionally, gradient SDF promotes robust accumulation of p53, with an associated increase in Ser15 phosphorylation and implicated kinases. We predict that reduced cell cycle progression through a p53 pathway supports cell motility in response to SDF gradient sensing. We also extend our findings on the Arf6‐ERK motility pathway and determined that it mediates invasion of metastatic breast cancer cells, and that Arf6 drives ERK to pseudopodia and invadopodia upon gradient SDF treatment. These results provide insight into the biochemical pathways downstream of CXCR4 gradient sensing, with implications on the invasive phenotype of metastatic breast cancer cells.

Supported by grant GM‐097718
The Functional Role of Arrestin Domain‐Containing Protein 3 in β2‐Adrenergic Receptor Endocytic Trafficking
Authors: Tian Xufan , Puthenveedu Manojkumar , Benovic Jeffrey
Published: June 2021

Arrestin domain‐containing protein 3 (ARRDC3) is a member of the mammalian α‐arrestin family, which includes ARRDC1‐5 and TXNIP. α‐arrestins, together with visual/β‐arrestins and VPS26‐related proteins, form the arrestin clan. While the biological function of a‐arrestins is relatively unexplored compared to β‐arrestins, ARRDC3 has been proposed to play a role in G protein‐coupled receptor (GPCR) trafficking. Here we use molecular, biochemical and cellular strategies to gain mechanistic insight into the role of ARRDC3 in GPCR trafficking. Our studies indicate that endogenous and expressed ARRDC3 primarily localizes on EEA1‐positive early endosomes, where it interacts with the ESCRT‐0 complex. ARRDC3 interacts also with the β2‐adrenergic receptor (β2AR) in a ligand‐independent manner, as observed by co‐immunoprecipitation and bioluminescence resonance energy transfer (BRET) analysis. While functional studies reveal that ARRDC3 has no effect on agonist‐promoted internalization or degradation of the β2AR, ARRDC3 colocalizes with internalized β2AR on early endosomes and significantly delays β2AR recycling back to the plasma membrane. This effect is not due to a general disruption in trafficking, since ARRDC3 has no effect on the recycling of the transferrin receptor. High resolution live cell imaging suggests that ARRDC3 negatively regulates β2AR localization in endosomal tubular protrusions. In conclusion, we propose that ARRDC3 interacts with the β2AR on EEA1‐positive early endosomes and negatively regulates β2AR‐containing endosomal recycling tubules.
mGluR1‐β‐arrestin 2 Signaling Mediates Induction of Excitatory Synaptic Plasticity
Authors: Eng Andrew , Hedrick Tristan , Swanson Geoffrey
Published: June 2021

A subset of seven‐transmembrane receptors (7TMRs) couple to both G protein‐ and β‐arrestin (βarr)‐mediated signaling pathways. Our objective is to determine if group I metabotropic glutamate receptors (mGluR1 and mGluR5) engage βarr signaling during the induction of synaptic plasticity in the CNS. In these experiments we tested the role of βarr‐Src‐MAPK signaling in the induction of plasticity at the hippocampal mossy fiber‐CA3 synapse.
We compared mGluR1‐dependent synaptic plasticity in patch clamp recordings from CA3 pyramidal neurons in hippocampal slices prepared from wildtype, βarr 1 and βarr 2 knockout mice. Potentiation of EPSCs induced by low frequency, paired stimulation of mossy fiber inputs to CA3 pyramidal neurons was absent in mice lacking βarr 2 (91 ± 7% n=9 for βarr 2‐/‐, 125 ± 9% n=18 for wildtype, p=0.01) but intact in mice lacking βarr 1 (128 ± 11% n=8 for βarr 1‐/‐, 122 ± 13% n=17 for wildtype, p=0.74). In contrast, high‐frequency stimulation that induced long‐term potentiation was unaffected by gene targeting of either βarr isoform (190 ± 23% n=9 for βarr 1‐/‐, 153 ± 18% n=6 for wildtype, p=0.22; 163 ± 18% n=7 for βarr 2‐/‐, 167 ± 14% n=6 for wildtype, p=0.89). Pharmacological dissection implicated ERK1/2 of the MAPK pathway (104 ± 8% n=6 for u0126‐treated slices, 139 ± 9% n=15 for vehicle slices, p=0.01), whereas the role of Src tyrosine kinases was equivocal (114 ± 8% n=9 for PP2‐treated slices, 139 ± 9% n=15 for vehicle slices, p=0.06).
These data reveal a novel signaling function for βarrs when coupled to mGluR1 and therefore yield insight into new strategies for therapeutic targeting of group I mGluRs, which in recent years has been pursued for a number of neuropathologies.
Research supported by R21 NS088916‐01 from NINDS to GTS.