bluuebunny/crossref_metadata_2025 huggingface dataset:
Epigenomic Studies of Facioscapulohumeral Muscular Dystrophy (FSHD)
Authors: Zeng Weihua , de Greef Jessica C , Chen Yen‐yun , Chien Richard , Kong Xiangduo , Winokur Sara T , Ball Alexander R , van der Maarel Silvere M , Yokomori Kyoko
Published: June 2021

Most cases of Facioscapulohumeral Muscular Dystrophy (FSHD) are linked to a repeat number contraction of D4Z4 sequences in the subtelomeric region of chromosome 4q (4qter), so‐called “4q‐linked” FSHD. However, the role of the D4Z4 repeats and molecular consequences of their contraction in the disease are still unclear. With chromatin crosslinking and immunoprecipitation (ChIP), we found that histone H3 lysine 9 trimethylation (H3K9me3), which is present in normal cells’ D4Z4, is lost in FSHD. This epigenetic change was also observed in the rare FSHD cases with no detectable D4Z4 contraction (i.e., phenotypic FSHD), but not in other muscular dystrophies. The H3K9me3 modification on other repeat sequences we studied remains intact in FSHD. These data suggest that H3K9me3 loss at 4qter D4Z4 can serve as a faithful diagnostic marker for FSHD, and that FSHD is a disease with “epigenetic abnormality”. We identified HP1γ and cohesin recruitment by H3K9me3 to D4Z4 in a cell type‐specific manner in normal cells. This binding is lost in FSHD, which may be related to the muscle‐specific pathogenesis of the disease. Furthermore, cohesin plays an active role in HP1γ recruitment to D4Z4. We hypothesize that heterochromatic D4Z4 repeats spread a silencing chromatin structure to other genomic regions through HP1γ/cohesin‐mediated long‐distance chromatin interactions. D4Z4 heterochromatin is disrupted in FSHD, which may deregulate downstream target genes resulting in muscular dystrophy.
Early Intervention by Targeting inhibitors of Multiple Signal Transduction Pathways in LPS induced Human PBMCs
Authors: Mendis Chanaka
Published: June 2021

Lipopolysaccharide (LPS) is an endotoxin that induces the secretion of pro‐inflammatory cytokines in many cells and has been known to induce septic shock in humans. This project is aimed at developing a rapid diagnostic method to combat the effects of LPS and eventually other toxins by utilizing signal transduction pathways induced by of LPS in Peripheral Blood Mononuclear Cells (PBMC) as a prototype module. We have identified a set of LPS specific genes in human PBMCs through micro array analysis and have identified central signaling pathways consisting a set of LPS specific genes. A thorough investigation of the newly identified common pathways allowed us to find key pathway components that may alter the expression pattern of a set of genes and may have the potential to act as diagnostic markers before symptoms and septic shock can take effect. We will further evaluate the alterations induced by specific inhibitors of the known gene expression profile of LPS by first using RT‐PCR and then confirm the gene expression pattern by carrying out protein expression analysis through ELISA. We believe that our work will allow us to better understand the complex interactions of multiple signal transduction pathways induced by LPS.
Novel signaling interactions between proteinase‐activated receptor 2 and Toll‐like receptors in vitro and in vivo
Authors: NHU QUAN MINH , Rallabhandi Prasad , Shirey Kari Ann , Teijaro John , Farber Donna , Netzel‐Arnett Sarah , Antalis Toni , Fasano Alessio , Vogel Stefanie
Published: June 2021

Toll‐like receptors (TLRs) and proteinase‐activated receptors (PARs) function as innate immune biosensors in mucosal epithelial cells (ECs). We previously reported the functional and physical interactions between TLR4 and PAR2, and more recently, between PAR2 and the TLR adapter, TRIF, and established a novel model of “receptor cooperativity” between PAR2 and TLR4. We hypothesized that intracellular signaling pathways utilized by TLRs and PAR2 would converge either cooperatively or non‐cooperatively when co‐engaged. We demonstrate herein the cooperation between PAR2 and TLR2, TLR3, or TLR4 for activation of NF‐κB‐dependent signaling in mucosal EC lines. In contrast, activation of PAR2 negatively regulated TLR3‐dependent antiviral pathway, blunting the expression of TLR3/IRF‐3‐driven genes, as well as activation of IRF‐3 and STAT1. Similarly, TLR4‐TRIF‐activated, IRF‐3‐driven genes were also attenuated by PAR2 co‐activation. Consistent with these in vitro observations, PAR2−/− and TLR4−/− mice, which were refractory to footpad edema induced by PAR2 agonist peptide, were protected from mouse‐adapted H1N1 influenza A virus‐induced lethality when compared to wild‐type mice. These data support and extend our recently described, novel model of PAR2‐TLR4 “receptor cooperativity” and highlight the complexity of signaling integration between heterologous innate immune biosensors. (Supported by NIH Grants T32‐AI‐07540 (QN), R37 AI‐18797 (SV), R01 DK048373 (AF), R01 HL084387, R01 DK081376, R01 CA098369 (TA), R01 AI50632S1 (JT), and R01 AI050632 (DF))
Zucker lean and fatty rat hepatocytes respond differently to insulin and retinoids in the expression of glucose and lipid metabolic genes
Authors: Li Rui , Chen Guoxun
Published: June 2021

We found that retinoids synergized with insulin to induce glucokinase gene (Gck) expression in primary hepatocytes, indicating their roles in modulating insulin sensitivity. Whether any change of this synergy exists in insulin resistant animals is unknown. The expression levels of Gck, Srebf‐1c, Pck1 and Fas in hepatocytes isolated from Zucker lean (ZL, insulin sensitive) and fatty (ZF, insulin resistant) rats in response to increasing concentrations of all trans retinoic acid (RA) with or without 1 nM insulin were analyzed by real time PCR. For Gck, RA started to induce its expression at 0.1 uM and 3uM, and synergized with insulin in ZL and ZF hepatocytes, respectively. The Srebf‐1c mRNA was induced by RA at 0.1 uM and insulin in ZL, but not ZF hepatocytes. The Pck1 mRNA was induced by RA at 0.1uM or higher in both ZL and ZF hepatocytes. Insulin‐mediated suppression of Pck1 expression was attenuated by RA at 3uM in ZL, but not ZF cells. Fas mRNA was not affected by RA in both ZL and ZF hepatocytes after 6 hours of treatment. Our results showed that ZL and ZF hepatocytes respond differently to insulin and RA treatments, suggesting alterations of RA responses in liver of insulin resistant rats.

Grant Funding Source: AHA
Effects of dietary conjugated linoleic acid (CLA) on intestinal absorption of vitamin A
Authors: Giordano Elena , Banni Sebastiano , Quadro Loredana
Published: June 2021

Conjugated linoleic acid (CLA) is a polynsaturated fatty acid of ruminant origin. We previously showed that CLA t10, c12 and CLA c9, t11 isomers elevate vitamin A levels in rodent liver. To investigate whether this hepatic vitamin A accumulation results from increased intestinal vitamin A absorption, we performed gavage experiments with wild‐type male mice. 12 hr prior the experiment, mice were injected with P‐407, a lipase inhibitor that blocks chylomicrons clearance. Next morning, they were given an oral bolus of all‐trans‐[3H] retinol (2× 106 cpm) and unlabelled all‐trans‐ retinol (6μg) in 100 μl of olive oil or olive oil + either CLA isomer (70:30, v:v). 3 hr after the gavage, mice were sacrificed to collect serum and tissues.

HPLC analysis of retinoids and [3H]‐counts/min revealed reduced retinoids levels in the intestine of mice gavaged with olive oil + either CLA isomer compared to those given olive oil. In serum, retinoids levels were not different among groups, but triglycerides were elevated in the olive oil group compared to mice given olive oil + either CLA isomer.

These results suggest that a single dose of dietary CLA could affect intestinal chylomicron assembly and/or secretion, as well as vitamin A incorporation in these particles. Further experiments are ongoing to elucidate the molecular mechanisms of this effect and the consequences of chronic dietary CLA intake on intestinal vitamin A absorption.
Elucidating the mechanism of vitamin A uptake in tumor‐prone, vitamin A‐sensitive tissues
Authors: Schulte Christina Elaine
Published: June 2021

Vitamin A (retinol) is not only essential for maintaining normal eye, brain, placenta, and skin function, but its active derivatives have powerful effects on cell proliferation and differentiation. The long‐term goal of this study is to understand how cancer incidence can be reduced through the optimization of vitamin A trafficking and signaling in tissues prone to tumor development. Retinol, the major circulating retinoid, travels to peripheral tissues in complex with retinol binding protein (RBP). Recently, a retinoic acid responsive protein with no known biological function, STRA6, has been identified as an essential transmembrane receptor that mediates retinol uptake in retinal epithelial cells. It conferred RBP binding to retinal cells transfected with a STRA6 expression vector, as well as increased cellular uptake of vitamin A. The goal of our current study is to examine the mechanism of RBP‐bound retinol uptake in mammary, prostate, and colon epithelial cells, which express abundant levels of STRA6. We will utilize siRNA to determine whether knockdown of STRA6 inhibits retinol‐RBP uptake as assessed by tracer assays and fluorescent microscopy, as well as the requirement of STRA6 for retinol‐mediated differentiation, apoptosis, and cell cycle arrest. These studies will lay the foundation for developing strategies for optimizing vitamin A status and retinol trafficking in tissues prone to tumors.

Grant Funding Source: NIH SR03CA128091‐0
Identification of Lactoferrin‐binding Proteins from Human Serum
Authors: Wood Jeremy , Elamin Ashraf , McAbee Douglas D.
Published: June 2021

The iron‐binding protein lactoferrin (Lf) present in blood originates in neutrophils and is cleared from the circulation by the liver, but the function of Lf during its sojourn in the blood remains obscure. Lf has been shown to bind the ferroxidase ceruloplasmin in serum but its possible interactions with other blood proteins has not been reported. In this study, we have purified and identified a set of Lf‐binding proteins (Lf‐BP) from human serum. Human sera were chromatographed on Lf‐agarose in 0.15 M NaCl, 20 mM NaPi, pH 7.2. Bound proteins were eluted with 0.5 M NaCl and separated by one‐ and two‐dimensional gel electrophoresis. Tryptic fragments of fractioned Lf‐BP were analyzed by MALDI‐TOF/TOF mass spectrometry and mass data were used to identify individual Lf‐BP, which included ceruloplasmin, immunoglobulin G, thrombin, vitronectin, and transthyretin. Chromatography of Lf‐BP on Sephacryl‐S200 suggested that individual Lf‐BP were not part of a larger complex. Finally, Lf‐BPs blocked binding of 125I‐Lf with asialoglycoprotein receptors on rat hepatocytes, suggesting that Lf in complex with Lf‐BP cannot bind this receptor. Thus, Lf‐BP may comprise a network of proteins that influence Lf's in vivo activities in blood. (Support: NIH DK‐61984)
Bioavailability of Zn from biofortified rice assessed in a Caco–2 cell model and in suckling rat pups
Authors: Du Xiaogu , Jou Ming‐Yu , Hotz Christine , Lonnerdal Bo
Published: June 2021

Biofortification is a sustainable method of increasing the micronutrient content of staple foods. We assessed the bioavailability of Zn from biofortified and conventional varieties of rice by two methods. Rice samples were digested with pepsin in vitro at pH 2 or pH 4, followed by pancreatin digestion at pH 7. Uptake of 65Zn from labeled rice samples into Caco‐2 cells was measured after 6 h. Rat pups were intubated with 65Zn‐labeled undermilled and polished conventional rice varieties and Zn biofortified rice. Pups were killed after 6 h and tissues were collected for radioactivity measurements. Means of radioactivity in Caco‐2 cells were 3.5±0.9% and1.9±0.2% after digestion at pH 2 and 4, respectively. Corresponding cellular Zn uptake ranged from 0.39 to 1.01 μmole Zn/g and 0.20 to 0.53 μmole Zn/g of cellular protein. In rat pups, means of radioactivity were 0.8, 5.4, 2.8, 10.7, 27.3, and 53.0 % in stomach, perfused small intestine, perfusate, liver, carcass, and cecum‐colon, respectively. Mean whole body (perfused small intestine, liver, and carcass) 65Zn uptake varied from 36% to 59%. Zn uptake in rat pups and by Caco‐2 cells after digestion at pH 2 were closely correlated for all rice varieties (r=0.53, p<0.05), and especially for polished rice (r=0.79, p<0.05). Zn uptake in rat pups and by Caco‐2 cells after digestion at pH 4 were closely correlated for under‐milled rice (r = 0.78, p < 0.05). Since uptake of Zn by rat pups has been shown to be closely correlated to results from human adults it is likely that this cell model can be used for screening of Zn bioavailability from staple foods. This study was supported by HarvestPlus.
Excess ATP7A reduces the copper content of the mammary gland in pregnancy and lactation but does not alter levels of plasma ceruloplasmin
Authors: Linder Maria C. , Fullriede Hendrik , Brennan Grant , Chu Yu‐Hsiang , Llanos Roxana , Michalczyk Agnes , Mercer Julian , Ackland Margaret Leigh
Published: June 2021

Two Cu pumps, ATP7A and B, are critical for Cu transport and homeostasis in mammalian metabolism. Mammary gland (MG) and placenta are somewhat unique in expressing both ATPases, and the question arises as to their separate functions. We hypothesized that in the lactating MG, ATP7B promotes entry of Cu into the milk, while ATP7A promotes efflux of excess Cu back into the blood. In lactating human breast we showed with confocal microscopy that ATP7A was localized in the basolateral membrane. It moved towards the basolateral membrane from the TGN in response to lactational hormones in the PMC42 cell breast model. In transgenic mice with human ATP7A expression 10–20 fold higher than endogenous mouse Atp7a, total Cu in the mammary gland, brain and kidney was reduced. Hepatic Cu and plasma ceruloplasmin were unchanged; however, plasma ferroxidase activity was significantly higher in the transgenic animals. In lactation and throughout gestation, MG had half as much Cu as the wild type, as did the milk. Uptake of 64Cu tracer by MG 1h after i.p. injection increased about 3‐fold from early to late gestation in the wild type but was significantly lower in the ATP7A overexpressing mice. These findings are consistent with the concept that ATP7A promotes “backflow” of Cu from mammary epithelial cells to the maternal blood, not just in lactation but also during gestational mammary gland development. Supported by USPHS Grant HD 46949.
Gymnasterkoreayne B exerts chemopreventive effects through regulation of Nrf2 and p53 cooperatively
Authors: Lee Saet Byoul , Kang Kyungsu , Pan Cheol‐Ho , Lee Jae Kwon , Jung Sang Hoon , Lee En‐Ha , Nho Chu Won
Published: June 2021

Phytochemicals prevent cancer either through blocking initiation by detoxification of carcinogens or through suppressing by apoptosis. In this study, we have identified a Gymnasterkoreayne B (GKB), a polyacetylene compound, isolated from Aster koraiensis as a major principle for a significant quinone reductase (QR) induction via Nrf2‐ Antioxidant response element (ARE) pathway. Furthermore, the induction of QR activity was increased by GKB in vivo. Interestingly, GKB induced apoptosis and p53 and p21 protein levels. In p53‐ or p21‐ knock out cells, Nrf2 activation and stabilization was decreased than wild type cells. However, apoptosis was induced by GKB in the knock out cells, and more induced in Nrf2 knock down cells. From the results, we can suggest that Nrf2 and p53 have a close relationship in cancer cell model induced by phytochemicals in terms of cancer chemoprevention. Taken together, we present here that GKB not only increases QR through Nrf2‐ARE pathway but also induces apoptosis through p53, and that it could be induced cancer chemoprevention effect by the cooperative regulation of both blocking and suppressing of cancer.

Grant Funding Source: KIST‐Gangneung Institute intramural research grant (2Z03300)
Inhibitory effects of dietary luteolin on transendothelial migration of monocytes and formation of lipid‐laden macrophages
Authors: Kim Dongshoo , Kim Minsoo , Kang Sang‐Wook , Kang Young‐Hee
Published: June 2021

In the development of atherosclerosis, the leukocyte transmigration begins with a multistep adhesion process that captures leukocytes from circulating blood. Oxidatively‐modified LDL (ox‐LDL) involved in development of atherosclerosis has been shown to facilitate leukocyte transmigration. Our previous study showed that flavone luteolin inhibited ox‐LDL‐triggered expression of cell adhesion molecules on vascular endothelial cells responsible for the leukocyte recruitment. This study investigated that luteolin disturbed scavenger receptor (SR) expression and retarded foam‐cell formation of THP‐1‐derived macrophages. In macrophages treated with 20 μg/mL ox‐LDL there was a heavy Oil red O staining, which was diminished by 10 μmol/L luteolin, repealing formation of lipid‐laden foam‐cells. Additionally, luteolin blunted uptake of 40 μg/mL fluorescent DiI‐labeled acetyl LDL in THP‐1‐derived macrophages. Such effects of luteolin entailed its blockade of macrophage SR induction. Furthermore, luteolin dampened TNF‐α‐elevated matrix‐degrading MMP‐9 secretion in THP‐1 cells, thus impeding their transendothelial migration. These results demonstrate that dietary luteolin was effective in curtailing SR‐mediated lipid‐laden macrophages and disturbing transendothelial migration of monocytes. Supported by the MKETD Project for Regional Strategic Planning.
Aged black garlic improves insulin sensitivity in fructose fed rats
Authors: Kim Jihye , Kim Jungin , Kang Minjung , Han Junghee , Kang Younghee
Published: June 2021

Insulin resistance is an important initiating pathogenic mechanism of type 2 diabetes mellitus. Aged black garlic was reported to exert hypoglycemic effect in animal model of type 2 diabetes. This study is aimed to investigate the effect of aged black garlic on insulin resistance in rats with insulin resistance induced by fructose. Whole garlic was heated at 55°C and 90% relative humidity for 60 min, at 70°C for 60 min, and then at 85°C for 24 hr and fermented at room temperature for 3 wk to produce aged black garlic. Male Sprague Dawley rats (n=18) weighing 100–120 g were fed diet containing 60% cornstarch and 60% fructose with and without 5% freeze‐dried aged black garlic for 10 wk. Body weight, serum glucose, insulin, cholesterol and triglyceride levels, and Homeostasis Model Assessment‐Insulin Resistance (HOMA‐IR) of fructose group were significantly increased compared with the control group (p<0.05). Body weight and serum insulin levels of aged black garlic group were not significantly different from the control and fructose group. Consumption of aged black garlic significantly reduced serum glucose, cholesterol and triglyceride levels and HOMA‐IR compared with fructose group. Thus we concluded that aged black garlic could improve insulin sensitivity in fructose‐fed rats. This study was supported by the Korean Ministry of Knowledge and Economy.
Age‐dependent changes in the sphingolipid composition of CD4+ T cell membranes and immune synapses
Authors: Huang Zhaofeng , Marko Melissa G. , Bunnell Stephen C. , Wu Dayong , Merrill Alfred H , Meydani Simin Nikbin
Published: June 2021

Optimal T cell activation requires the formation of an immunological synapse (IS) and is driven by signaling ‘microclusters’ that associate with ordered membrane microdomains. Aging is associated with impaired IS formation and T cell activation. We hypothesized that age‐dependent changes in T cell lipid profiles impair IS formation and T cell activation in the aged. Total cell membrane (TCM) and IS fractions were prepared from the CD4+ T cells of young and old mice and subjected to ‘sphingolipidomic’ analysis. Both TCM and IS of aged CD4+ T cells have significantly higher levels of sphingomyelins and ceramides than young T cells. A subset of the lipids containing C14, C18:1, C26, and C26:1 fatty acid (FA) species was preferentially enriched in the IS. Young IS were enriched in C14 ceramide, C18:1 ceramide, C26:1 ceramide, and C14 hexosylceramide. Since sphingolipids are implicated in T cell activation and function, age‐dependent changes in the FA chain lengths and sphingolipid species retained in IS could influence T cell activation and function and may contribute to functional T cell defects in the aged.

Supported by NIA R21‐AG030931, R01‐AG009140‐14, Office of Dietary Supplement, USDA‐ARS contract number 58‐1950‐7‐707, and LIPID MAPS Consortium.
MODERATE EXERCISE AND ITS EFFECT ON THE SYSTEMIC IMMUNE RESPONSE ASSOCIATED TO HIGH FAT OR HIGH CARBOHYDRATE DIETS
Authors: Valdés‐Ramos Roxana , Martínez‐Carrillo Beatriz E , Aranda‐González Irma I , Guadarrama‐López Ana L , Jarillo Rosa Adriana
Published: June 2021

High Carbohydrate (HCD) or high Fat (HFD) diets produce negative effects particularly on the immune system. Moderate exercise may be able to modulate these negative effects. Our objective was to evaluate the effect of moderate exercise on the peripheral immune system, associated to HCD or HFD intake. Three groups (n=8 each) of 21 day‐old Balb/c mice were fed control, HCD or HFD diets. During their fourth week of life, they were trained to swim with time increasing sessions (10, 15, 20, 25 and 30 minutes), after which they continued swimming for eight more weeks at 30‐minute sessions, five days a week with two days of rest. At 12 weeks of age, blood was obtained by intracardiac puncture for lymphocyte separation and analysis by flow cytometry (CD4+, CD8+ and CD19+). CD4+ lymphocytes were non‐significantly higher in HCD (49.68±3.01) and HFD (57.51±7.43), compared with controls (47.88±11.34). CD8+ were significantly higher (p<0.0001 and F=29.33) in the experimental diets (HCD =18.30±4.27; HFD 16.58±2.48) than controls (8.64±3.11). On the other hand, CD19+ were higher in the control group (18.46±6.23) than HCD (16.67±3.98) and HFD (13.16±3.89), p <0.001 and F =5.49. Moderate physical exercise associated with a HCD mainly increased T‐cytotoxic (CD8+) lymphocytes, whereas together with HFD it increased T‐helper (CD4+) cells. Both types of diets decreased B lymphocytes (CD19+). This project was financed by UAEM.
100% Tart cherry juice reduces pro‐inflammatory biomarkers in overweight and obese subjects
Authors: Martin Keith R , Burrell Lacey
Published: June 2021

Subclinical inflammation is frequently noted in chronic disease including diabetes, cardiovascular disease (CVD) and obesity. Accumulating epidemiological evidence shows that diets rich in fruits, e.g., cherries, may significantly reduce risk, in part, via antioxidant and anti‐inflammatory activities. In this randomized, placebo‐controlled crossover study, we recruited 10 participants (38.1 ± 12.5 y; 8 females, 2 males) with BMI>25.0 (32.2 ± 4.6; 5 obese, 5 overweight) to consume 8 fl. oz. daily of either 100% tart cherry juice or an alternate placebo beverage, for 4 weeks each with a 2 week intervening washout period. Fasting blood samples were collected at the beginning and end of each arm for measurement of biomarkers of inflammation. The erythrocyte sedimentation rate (ESR), an indicator of chronic inflammation, was significantly (p<0.05) lower with treatment than with the placebo beverage. Baseline hsCRP, an indicator of acute inflammation, was 7.0 ± 5.2 mg/l indicating increased CVD risk in these participants. Consumption of cherry juice did not affect hsCRP levels. The chemokine MCP1 and cytokine TNF‐alpha were lower (p<0.05) in participants that started the trial with cherry juice compared to the placebo beverage. Plasma IL‐6 and IL‐10 were not different between treatments. Collectively, the data suggest that 100% tart cherry juice may reduce biomarkers of inflammation often noted in chronic disease.
Curcuma or citrus extract supplementation does not improve body condition nor insulin sensitivity in obese cats
Authors: Leray Veronique , Jeusette Isabelle , Serisier Samuel , Freuchet Benjamin , Torre Celina , Nguyen Patrick
Published: June 2021

The putative therapeutic properties of curcuma and citrus could be associated with their antioxidant and anti‐inflammatory properties. Our aim was therefore to examine the effects of citrus or curcuma extract supplementation on body condition and inflammatory state in obese cats.

Eight obese cats participated in this study that was conducted as a cross‐over, with two 8‐week periods. The diets were supplemented with either curcuma or citrus extract. Body composition (D2O dilution method), insulin sensitivity (euglycemic hyperinsulinemic clamp technique), plasma leptin level (RIA), and TNFα, IL1β and IFNγ mRNA expression in leucocytes (real‐time PCR) was assessed prior to and at the end of the test period.

Body weight was unchanged by either diet. Both diets lowered fat‐free mass whereas the percentage of fat mass increased. Basal glycemia, basal insulinemia, insulin sensitivity and plasma leptin level were not modified whatever the diet. TNFα, IL1β and IFNγ mRNA levels were not affected by either diet.

Similar plasma leptin level in both groups is consistent with their body weight and body composition. Our results suggest that curcuma or citrus extract supplementation is not effective in improving body weight, body composition, insulin sensitivity, or inflammatory state in obese cats.
Biological activity of avocado‐derived mannoheptulose in dogs
Authors: Davenport Gary , Massimino Stefan , Hayek Michael , Burr John , Ceddia Michael , Yeh Chyon‐Hwa , Roth George , Ingram Donald
Published: June 2021

Mannoheptulose (MH) is a 7‐carbon sugar found in avocados and other natural sources. It is proposed as a calorie restriction (CR) mimetic that delivers anti‐aging and health‐promoting benefits of CR without reducing food intake. Two studies were conducted to evaluate effects of MH on fasting insulin when fed as an avocado extract (AvX) to Labrador Retrievers (LR) and Fox Terriers (FT). Decreased fasting insulin is commonly associated with CR and longevity. In Study 1, 36 dogs, balanced by breed (18 LR; 18 FT) and age (18 young, 2.6 yr; 18 old, 11.8 yr), received control or test treatments during sequential 2‐week periods. Test dogs were fed one AvX‐containing gelatin capsule daily to deliver 1, 2, 10 or 20 mg MH/kg BW in a step‐up fashion. Control dogs were fed one placebo capsule daily. Fasting blood samples were collected weekly. Fasting insulin was lower (P<0.05) in dogs fed 2, 10 and 20 mg/kg. In Study 2, LR (15) and FT (15) were fed an AvX‐containing diet providing 0, 2 or 5 mg MH/kg BW. A glycemic meal challenge was induced on days 0 and 30 by feeding dogs one‐half their daily meal allotment after an overnight fast. Blood samples for insulin and glucose were collected at 0, 30, 60, 120 and 240 min post‐feeding. MH (2 mg/kg) reduced (P<0.05) insulin levels (mean, peak, area under curve) in old LR on day 30. Glucose was not affected by MH. Lower plasma insulin implies mannoheptulose is biologically active in dogs when fed as an avocado extract.
Effect of Methyl Nutrients on Feline Lymphoma Growth
Authors: Mabasa Lawrence , Cho Kyongshin , Fowler Andrea W. , Park Chung S.
Published: June 2021

Feline lymphoma is one of the most frequently diagnosed tumors in cats. Lipotropes are dietary methyl donors and cofactors that are involved in one‐carbon metabolism and DNA methylation. However, little is known about their role in feline lymphoma. We hypothesized that supplementation of lipotropes would modulate the DNA methylation status and the expression of genes involved in cell growth and apoptosis of feline lymphoma cells. The objective of the study was to determine if lipotropes affect growth of feline lymphoma cells: entailing examining a correlation between lymphoma cell growth and caspase‐3 enzyme activity. Feline lymphoma cell lines, F1B and FeLV‐3281, were cultured and treated with 20 times the level of each lipotrope contained in the basal culture medium (control). Cell growth and caspase‐3 activity were measured. Lipotropes significantly reduced cell growth (45.4%) and increased caspase‐3 activity (4.9%, P = 0.01) of F1B cells after 72 hours. However, cell growth and caspase‐3 activity of FeLV‐3281 were not affected by lipotropes. These results suggest that methyl nutrients may play a role in reducing cell growth of F1B, but not FeLV‐3281. Further studies are warranted to determine the mechanism underlying the inhibitory effect of lipotropes on F1B lymphoma cell growth.

Supported by Morris Animal Foundation (D08FE‐047).
Differential activation of NF‐κB in colonic mucosa of DSS‐challenged rats consuming fermentable fiber sources
Authors: Turner Nancy D. , Taddeo Stella S. , Callaway Evelyn S. , Fan Yang Y. , Davidson Laurie A. , Thomas Laura N. , Ferguson Carly M. , Sturino Joseph M. , Chapkin Robert S. , Lupton Joanne R.
Published: June 2021

NF‐κB activation is associated with colon inflammation. We compared dietary fiber effects on SCFA, microbiota and NF‐κB activation in a colitis model. Rats received diets containing corn oil and: 6% cellulose (CELL); or 2% CELL with 4% of PROMITOR™ soluble corn fiber (SC), PROMITOR™ resistant starch (RS), or SC+RS (MIX) for 2 wk before DSS (3%, 2×, 14 d separation) (n=20/diet) or control treatment (no DSS, n=10/diet). Tissues were collected 10 d after DSS for NF‐κB activity, injury and inflammation scores. Feces were analyzed for SCFA (μM/g) and microbiota (qPCR 16S rRNA). DSS elevated SCFA (P<0.03), except for CELL. Butyrate concentration was not affected by DSS, yet % butyrate decreased in DSS rats (P<0.05 for CELL and RS). Enhancement of NF‐κB activity by DSS was 25, 50, 61, and 32% for the CELL, SC, RS, and MIX diets, significant for all but CELL. Injury scores were not affected by CELL, but were >75% higher with SC and MIX. Changes in microbiota occurred as rats transitioned from a pelleted diet to experimental diets. Bifidobacteria numbers were elevated with the RS diet. These data suggest in rats consuming n‐6 PUFA, that SCFA production from altered microbiota is correlated with NF‐κB activity in this colitis model. It is important to elucidate the underlying mechanisms to appreciate how dietary fiber may influence colon inflammation. Support: Tate & Lyle Health & Nutritional Sciences.
Dietary macronutrients modify serum metabolomics in rats
Authors: Kristal Bruce , Baranov Sergei , Stavrovskaya Irina , Greenberg Heather , Sheldon Diane , Marur Vasant , Russell Neil , Sniatynski Matt , Porter Caryn
Published: June 2021

Suboptimal dietary macronutrient choices are arguably the major environmental stressor in individuals living in Western societies. Trans and saturated fats contribute to cardio‐ and cerebrovascular disease, as do diets high in easily digested carbohydrates, which themselves may contribute to metabolic syndrome and overt diabetes. A massive literature links diet and both neoplastic and non‐neoplastic diseases, and a causal role for mitochondria in this relationship is strongly supported by broad areas of inquiry represented in the literature. We are funded as a part of the NIH‐sponsored Genes and Environment Initiative, Exposure Biology Program (Biological Response Indicators component) to conduct an interdisciplinary approach to discover and confirm plasma metabolomic biomarkers for dietary intake of subclasses of fats and carbohydrates and their effects on mitochondrial function. Metabolomics measurements in sera/plasma were conducted by HPLC w/Coularray detectors. Fisher‐Brown Norway F1 rats were fed one of twenty‐four different diets systematically differing in fats (saturated, trans, MUFA, and 3 w‐6/w‐3 ratios crossed with 4 levels of glycemic index). We will present the initial effects of these diets on the sera metabolome. Future studies will investigate these biomarkers in nested human case control studies. NIEHS funding.
Dietary fish oil decreases DNA adducts in the rat colon independent of estrogen status
Authors: Armstrong Cameron , Allred Clinton D.
Published: June 2021

Colon cancer is the third leading cause of cancer related death in men and women in the United States. Data from multiple studies show that dietary fish oil is associated with a decrease in colon cancer incidence. Estradiol (E2) has also been demonstrated to be protective against colon tumor formation and we have shown that E2 induces colonocyte apoptosis in conjunction with reducing formation of premalignant lesions. To date, however, possible relationships between estrogen status and the benefits of fish oil have not been thoroughly examined. We hypothesized that E2 could enhance the effects of dietary fish oil by additively decreasing DNA damage and increasing apoptosis. Dietary fish oil significantly decreased immunohistochemical staining intensity of O6‐methyldeoxyguanosine DNA adducts independent of the presence of E2 at 9h and 12h post treatment with carcinogen. E2 resulted in a significant increase in apoptosis independent of diet group at 12h post carcinogen, most notably in the luminal third of the crypts. Fish oil was not associated with an increase in apoptosis. These data are important because they demonstrate that fish oil is protective against DNA damage in the colon regardless of E2 status. In addition, our data demonstrate that induction of apoptosis in colonocytes at the point of DNA damage may serve as a primary mechanism for how E2 suppresses colon tumor formation.

Grant Funding Source: American Institute for Cancer Research
DHA Exacerbates Experimentally Induced Colitis in SMAD3−/− Mice
Authors: Woodworth Hillary L , McCaskey Sarah J , Clinthorne Jonathan , Langohr Ingeborg , Duriancik David , Gardner Elizabeth M , Fenton Jenifer I
Published: June 2021

Inflammatory bowel disease (IBD) increases the risk of developing colorectal cancer. It is hypothesized that dietary interventions can reduce inflammation and associated cancer risk. The long chain omega‐3 fatty acid, docosahexaenoic acid (DHA) has potent anti‐inflammatory properties. The objective of this study was to determine whether dietary DHA could reduce experimentally induced colitis and subsequent colon cancer risk. We utilized a mouse model of colitis and colon adenocarcinoma formation (SMAD3−/−). When SMAD3−/− mice are exposed to H. hepaticus, colitis is observed 4 wks post infection. Mice (10 per group) were fed AIN‐93G powdered diets supplemented with corn oil, safflower oil, or DHA‐rich fish oil (doses ranging from 0.75–6%) for 8 wks. Mice were then gavaged with H. hepaticus, continued on their diets and sacrificed 4 weeks post‐infection. Colon and cecal tissue were collected for histopathology and scored for inflammation and dysplasia. Spleen, peyers patch and mesenteric lymph nodes were collected for CD3+ cell populations. Contrary to expectations, DHA (2.25–6%) induced severe colitis and adenocarcinoma formation. Increased severity of colitis in DHA fed mice was associated with a CD8+ driven response to bacterial infection. These results suggest that DHA supplementation in immune‐associated diseases like IBD should be approached with caution.
Relationships between folate status, genetic polymorphisms in the methylenetetrahydrofolate reductase and thymidylate synthase genes and susceptibility to hepatocellular carcinoma
Authors: Kuo Chang Sheng , Huang Rwei‐Fen Syu
Published: June 2021

The aims of the study were to investigate interactive relationships between serum folate levels, MTHFR and TS genetic polymorphisms and risks of HCC development. Genotypes of MTHFR C677T, a 28‐bp tandem repeat in the TS 5′‐untranslated enhanced region, and a 6‐bp deletion/insertion in the 3′‐untranslated region were determined on 190 HCC cases and 190 healthy control subjects. Serum samples were assayed for folate and homocysteine (Hcy) concentrations. The data revealed that serum folate of HCC patients was lower and Hcy concentrations were higher than those of controls (P < 0.05). The distributions of CT/TT genotypes significantly differ between HCC patients and controls (P = 0.018). Subjects with the CT/TT genotype had a decreased risk for HCC relative to those with CC genotype, particularly among individuals with adequate folate status (serum folate > 14 nmol/L). Relative to those with low folate status, individuals with adequate folate status had a significant 80–90% reduction in risk of HCC. The MTHFR CT/TT genotype in conjunction with TSER or TS 3′UTR polymorphisms was associated with a statistically nonsignificant 50–60% reduction in risk of HCC as compared with the CC genotype. Taken together, our data supports the hypothesis that adequate folate status in combination with at least one mutant allele in MTFHR C677T or/and TSER or TS 3′UTR genotype may protect against the development of HCC.
Effect of Curcumin on BOP Induced Oxidative Stress in Hamster Model: Correlation with Metabolomic Profile
Authors: Zaheeruddin Shaistha , Goja Arvind , Gupta Smiti
Published: June 2021

Oxidative stress (OS) is associated with many chronic diseases including cancer, CVD and the normal process of aging. Curcumin, a dietary phytochemical has proven antioxidant, anti‐inflammatory and anti‐carcinogenic activity. This study aims to utilize metabolomics to evaluate a signature of oxidative stress in BOP (N‐nitrosobis‐2‐(oxopropyl) amine) treated hamsters. Further, the effect of dietary curcumin on the change in metabolomic signatures was investigated. 32 male Golden Syrian hamsters were randomized into treatment (n=20; OS induced by BOP), and control (n=12) groups. After 12 weeks each group was subdivided into no (T0, C0) or 2% curcumin (T2, C2) diet groups. Proton nuclear magnetic resonance spectroscopy was conducted on 24h urine samples, in Deuterium oxide using a 700 MHz instrument at 25°C. The NMR spectra were processed and analyzed using Principal Component Analysis (PCA; SIMCA P+, Umetrics), an unsupervised multivariate analysis technique. The PCA score plots showed clear separation between spectra from (i) the treatment (T0) and control (C0) groups (ii) groups with (T2) and without (T0) dietary curcumin (p < 0.05). The PLS (Partial Least Square) analysis was used to correlate the metabolomic profiles with Total Antioxidant levels, lipid peroxide levels, cyclooxygenase‐2 (COX‐2) activity and NF‐κB activity. The metabolites responsible for the inter‐group differences were quantified using the CHENOMX metabolite database. These metabolites, in addition to providing a mechanistic insight, may serve as potential signatures of oxidative stress.
Strategies to Engage the Next Generation of Scientists
Authors: Gentile Lisa N
Published: June 2021

I will discuss four programs that I am involved with to encourage students to pursue futures in the sciences. The first, funded by NSF‐CAREER, is one that encourages high school students (some high achieving and some from more disadvantaged programs) and their teachers to spend their summers doing research and designing experiments to bring back to their classrooms. Examples of research projects, and outcomes as well as experiments designed will be presented. Next, an integrated, quantitative approach to teaching introductory science/math education, funded by the Howard Hughes Medical Institute (HHMI), will be described. Having just been designed and implemented, results from the first year of the program will be shared. Strategies of an undergraduate research program, funded by NSF‐CAREER, HHMI, and MERCK‐AAAS, focused on bringing students into productive long term research projects, as early as the summer prior to their freshman year, will also be discussed. Finally, the benefits of including post‐baccalaureate students into undergraduate research programs will be detailed, both from the perspective of the students involved as well as the forward momentum of the lab.
Water turnover assessment in overweight adolescents
Authors: Weinheimer Eileen , O'Connell Bláthnaid N , Martin Berdine R , Schoeller Dale A , Weaver Connie M , Campbell Wayne W
Published: June 2021

Adequate intake (AI) standards for water (H2O) in adolescents range between 2.4 – 3.3 L/d for males and 2.1 – 2.3 L/d for females, independent of obesity status. Water intakes and excretions of this population are not well documented.

Purpose

To assess H2O turnover, inputs and outputs in overweight adolescents.

Methods

18 girls (BMI 31.7 ± 1 kg/m2; mean ± SEM) and 9 boys (BMI 26.3 ± 1 kg/m2) aged 12–15 y completed two 3‐wk metabolic balance trials. Rate of H2O turnover (rH2O) was measured by tracking the decline of deuterated H2O from the body over 14 d. Water inputs (diet*, ad libitum#, metabolic#) and outputs (urine*, feces*, insensible#) were assessed (*=measured, #=estimated).

Results

Using both trials (girls n = 36 trials; boys n = 18 trials), rH2O was higher (p<0.05) in boys vs. girls (4537 ± 202 vs. 3742 ± 122 g/d). Per kg body weight, rH2O was 28% lower in girls vs. boys (46 ± 7 vs. 64 ± 9 g kg−1 d−1). Water input from diet, ad libitum intake and metabolic production were 44, 19, and 28% lower, respectively, in girls vs. boys. Urine, fecal and insensible H2O losses were 29, 17, and 29% lower in girls v. boys. BMI did not influence rH2O. The intraclass correlation of rH2O between trials was 0.981 (p < 0.001).

Conclusion

These overweight adolescents consumed H2O well in excess of sex specific AI standards. The higher rH2O in boys vs. girls is consistent with adult males vs. females.
Convenience Drives Choice in School Lunch Rooms: A Salad Bar Success Story
Authors: Smith Laura E. , Just David R. , Wansink Brian
Published: June 2021

Efforts to improve food choice in high schools and middle schools often focus on either taxing or on eliminating less healthy alternatives ‐‐ both which could drive students away from the school lunch program. Building on the premise that two influences on school lunch decisions are salience and convenience, a field was conducted in a middle school where the objective was made to make the salad bar more salient and convenient to students. Instead of being placed parallel to a lunchroom wall, it was moved 4 feet and rotated 90 degrees perpendicular to the cash registers. Depending on the specific food items, the resulting salad sales increased 300 to 400 percent compared from year‐to‐date figures. Additionally, these sales occurred without a decrease to overall lunchroom sales or per/person expenditures. A variety of additional “healthy‐foods‐made‐more‐convenient” opportunities are suggested as next‐step interventions worth of investigation.

Grant Funding Source: none
A 10 week‐lifestyle modification program reduced the metabolic syndrome in overweight adults
Authors: Burini Roberto Carlos , Mecca Marita Scarpim , Moreto Fernando , Burini Franz Homero Paganini , Pierine Damiana Tortolero
Published: June 2021

PURPOSE

To reduce metabolic syndrome (MetS) after a short‐time (10 wk) lifestyle modification program (LMP) with physical exercises and low‐energy density diet (30g fiber/day).

METHODS

In a controlled parallel group design participants (n=50; 50.5 ± 11.8 yrs) were allocated either to intervention with adequate intakes of fruits and vegetables (G1, n=28, 6M, 22F, BMI=33.6 ± 5.3kg/m2, MetS=46%) or control (G2, n=22, 5M, 17F, BMI=32.1 ± 5.3kg/m2, MetS=40%). Both groups were in a weekly LMP with 60 min/d sessions of aerobic exercises alternating brisk walking (3d/wk) and muscle strength (2d/wk). Clinical, anthropometric, dietary, fitness and plasma biochemical assessments were undertaken at baseline (M0) and after 10 weeks (M1). Groups and moments were statistically compared for p=0.05.

RESULTS

At M0 both groups were similar in all variables and after 10 weeks of LMP they both improved health‐eating index (HEI), fiber intake and kept their muscle mass. However only G1 increased the intake of fiber (211%) along with plasma beta‐carotene (150%), additionally to greater reductions in body fat (4%), abdominal fat (7%) and obesity (16%) leading to reduced MetS (24%) mainly by normalizing blood pressure, triglycerides and glucose.

CONCLUSIONS

Short‐time (10 weeks) LMP reduced MetS by improving HEI, body composition and mostly blood markers. Supported by CNPq and FAPESP.
Alternate day fasting: A novel dietary strategy for weight loss and cardio‐protection in obese adults
Authors: Varady Krista A , Bhutani Surabhi , Church Emily C , Klempel Monica C
Published: June 2021

This study examined the effects of alternate day fasting (ADF) (i.e. consuming 25% of energy needs on the fast day, and ad libitum fed on feed day) on body weight and coronary heart disease (CHD) risk in obese adults. Sixteen obese subjects (12 women/4 men) completed a 10‐week trial, consisting of 3 phases: 1) 2‐week control phase, 2) 4‐week weight loss/ADF controlled feeding phase, and 3) 4‐week weight loss/ADF self‐selected feeding phase. Dietary adherence remained high throughout the controlled feeding phase (86% days adherent), and the self‐selected feeding phase (89% days adherent). Rate of weight loss remained constant during controlled feeding (0.67 ± 0.1 kg/week) and self‐selected feeding phases (0.68 ± 0.1 kg/week). Body weight decreased (P < 0.001) by 5.6 ± 1.0 kg (5.8% ± 1.1%) after 8 weeks of diet. Percent body fat decreased (P < 0.01) from 45 ± 2% to 42 ± 2%. Total cholesterol, LDL cholesterol, and triacylglycerol concentrations were lowered (P < 0.01) by 21 ± 4%, 25 ± 10%, 32 ± 6%, respectively, after 8 weeks of ADF, while HDL cholesterol remained unchanged. Systolic blood pressure decreased (P < 0.05) from 124 ± 5 mm Hg to 116 ± 3 mm Hg. These findings suggest that ADF is a viable diet option to help obese individuals lose weight and decrease CHD risk.
High protein diets modulate renal lipid metabolism in Zucker fa/fa rats
Authors: Tovar‐Palacio Clauida , Tovar Armando R , Torres Nimbe , Cruz Cristino , Hernandez‐Pando Rogelio , Salas‐Garrido Gerardo , Correa‐Rotter Ricardo
Published: June 2021

It is not clear whether the use of high protein diets during obesity affects renal lipid metabolism and kidney function.

Thus, the aim of this study was to assess the effect of different type and amount of dietary proteins on the regulation of the expression of lipogenic and inflammatory genes and its association with renal lipid concentration and biochemical parameters of kidney function in obese Zucker fa/fa rats. Rats were fed for 2 mo different concentrations of soy protein (SP) or casein (Cas) (20, 30, 45%). SP significantly increased serum TG than Cas; independently of the type of protein, the higher the dietary protein concentration, the higher the serum TG. Rats fed SP significantly decreased serum cholesterol concentrations than those fed Cas diet. No significant changes in renal TG and cholesterol concentrations were observed. Renal gene expression of SREBP‐1 and SREBP‐2 expression and their target genes showed a significant increase as the concentration of dietary protein increased. The higher the protein concentration the higher the expression in the kidney of the proinflammatory cytokines, independently of the type of dietary protein. In addition, a similar response was observed for nefrin, Type IV collagen, and TGFβ. These results indicate that dietary protein regulates expression of lipogenic and proinflammatory genes in kidney regardless of its type. Supported by CONACYT No 49098 (to CTP).