bluuebunny/crossref_metadata_2025 huggingface dataset:
Mono‐2‐ethylhexyl phthalate exposure impairs embryonic growth and nutrition and alters epigenetic programming during early organogenesis (137.8)
Authors: Sant Karilyn , Jilek Joseph , Dolinoy Dana , Harris Craig
Published: June 2021

The early organogenesis stage of development provides a window of susceptibility during which the nutrient environment guides processes such as epigenetic programming, cellular differentiation, and overall growth. The placenta is not yet functional, leaving receptor mediated endocytosis (RME) of proteins and their cargoes through the visceral yolk sac as the primary source of embryonic (EMB) nutrition. Supplementation of maternal diets with methyl donors and cofactors required for one‐carbon metabolism (C1) has been shown to modify epigenetic programming and prevent neural tube defects. Environmental agents, such as the plasticizer MEHP, are associated with abnormal growth and neurodevelopment, yet the mechanistic relationship between chemical and nutritional agents remains unknown. Our objective was to assess whether MEHP impairs growth, attenuates embryonic nutrient uptake, and alters epigenetic programming during organogenesis.

Mouse conceptuses were explanted into whole embryo culture (WEC) on GD 8 and exposed directly to 100 or 250 μg/ml MEHP. After 24 h, exposed embryos showed a dose‐dependent increase of incomplete closure of the neural tube, increased cellular oxidation, and produced a significant reduction in overall growth. Assessment of RME activity, based on FITC‐albumin uptake, showed dose‐dependent decreases in histiotrophic nutrition. Conceptal C1 substrates were measured after 24 hr using mass spectrometry with vitamin B12, choline, and homocysteine being decreased only at the low dose. Betaine and folate were decreased in both the low‐ and high‐dose samples while methionine, cysteine, and S‐adenosylmethionine increased with dose. Histone hypermethylation due to treatment occurred at H3K27 loci, while hypermethylation of H3K4 occurred only in EMB with open neural tubes. This work demonstrates the ability of environmental chemicals to modify EMB nutrition and epigenetic programming, and shows that these changes may lead to growth dysregulation.

Grant Funding Source: Supported by NIEHS (P30 ES017885, T32 ES07062) and the Bill & Melinda Gates Foundation (Round 7).
Leukocyte specific protein‐1: a novel regulator of hepatoma proliferation and migration (144.10)
Authors: Koral Kelly , Paranjpe Shirish , Mars Wendy , Michalopoulos George
Published: June 2021

Copy number variation analysis of human hepatocellular carcinoma samples revealed leukocyte specific protein‐1 (LSP1) gene had the greatest number of deletions. The role of LSP1 in the development or progression of hepatocellular carcinoma remains unknown therefore our objective was to determine the functional significance of LSP1 in liver cancer. Using shRNA against LSP1, we were able to create a stable cell line using the LSP1 expressing JM1 rat hepatoma cells. Analysis of proliferation and migration utilizing the MTT and scratch assay, respectively revealed that loss of LSP1 expression leads to increased proliferation and migration in the JM1 stable cell line as compared to scrambled shRNA control. The stably expressing LSP1 shRNA JM1 cells expressed higher levels of the proliferation markers, cyclin D1 and PCNA providing further evidence of a role for LSP1 in regulating the proliferation of hepatoma cells. Future studies will aim to examine if expression of LSP1 in a LSP1 deficient cell line will result in decreased proliferation and migration. Understanding the function of LSP1 in liver cancer may lead to novel therapeutic targets to combat this deadly disease.

Grant Funding Source: Supported by NIH
Therapeutic induction of arteriogenesis in humans (19.4)
Authors: Seiler Christian
Published: June 2021

Coronary collaterals are an alternative source of blood supply to myocardium jeopardized by ischemia. In comparison to other species, the human coronary collateral circulation is very well developed. Collateral arteries preventing myocardial ischemia during a brief vascular occlusion are present in 1/3 of patients with coronary artery disease (CAD). Among individuals without coronary stenoses, there are preformed collateral arteries preventing myocardial ischemia in 20‐25%. Determinants of preformed coronary collaterals are low heart rate and the absence of systemic arterial hypertension. Collateral flow sufficient to prevent myocardial ischemia during coronary occlusion amounts to 1/5th to 1/4th the normal flow through the open vessel. Myocardial infarct size, the most important prognostic determinant after such an event, is the product of coronary artery occlusion time, area at risk for infarction and the inverse of collateral supply. Well developed coronary collateral arteries in patients with CAD mitigate myocardial infarcts and improve survival.

Approximately 1 out of 5 patients with CAD cannot be revascularized by percutaneous coronary intervention or coronary artery bypass grafting. Therapeutic promotion of collateral growth is a valuable treatment strategy in those patients. Promotion of collateral growth should aim at inducing the development of large conductive collateral arteries (i.e. arteriogenesis). They appear to be effectively induced by activation of monocytes / macrophages or by physical augmentation of coronary blood flow velocity.
Comparison of dietary supplement intake reporting between the Automated Self‐Administered 24‐Hour Recall (ASA24) and Automated Multiple Pass Method (AMPM) recalls (245.2)
Authors: Schap TusaRebecca , Thompson Frances , Dodd Kevin , Potischman Nancy , George Stephanie , Coleman Laura , Gahche Jaime , Alexander Gwen , Groesbeck Michelle , Kahle Lisa , Kushi Lawrence , Clancy Heather , Sundaram Maria , Kirkpatrick Sharon , Dixit‐Joshi Sujata , Zimmerman Thea , Douglass Deirdre , Mittl Beth , Bailey Regan , Subar Amy
Published: June 2021

Objective: To compare dietary supplement use reported on self‐administered (ASA24) versus interviewer‐administered (AMPM) 24‐hour recalls.

Methods: About 1200 women and men were recruited from three integrated health systems using quota sampling to capture a range of ages and racial/ethnic backgrounds. Participants were randomized into four groups and asked to complete two recalls: 1) two ASA24s; 2) two AMPMs; 3) ASA24 first, AMPM second; 4) AMPM first, ASA24 second. Dietary supplements were coded using the 2007‐2008 NHANES Dietary Supplement Database. Difference in prevalence of reported supplement use was tested using a Chi‐square test.

Results: Food and supplement data were complete for 1089 participants. Prevalence of use of any dietary supplement on either day was not significantly different by recall mode overall (47.8% for AMPM (n=1022) vs.48.5% for ASA24 (n=1012); p=0.82) or within sex, age, or race/ethnicity groups. Among supplement users, the prevalence of reported multivitamin use was significantly higher on AMPM compared to ASA24 (65.9% vs. 57.2%; p <0.01). In ASA24, 50 participants reported 114 supplements as “unknown”, meaning they were unable to find their supplement in the ASA24 database.

Discussion: The ASA24 performs similarly to AMPM for estimating overall prevalence of dietary supplement use. Significant differences found in the reporting of multivitamins specifically could be related to the number of unknown supplements reported in ASA24. Coding of text entered for supplements not found in ASA24 may be necessary to obtain accurate prevalence of supplement type and nutrient estimates. Based on these findings, future versions of ASA24 will include questions when unknown supplements are reported for the purpose of assigning reasonable default supplement codes.
Blood glucose lowering effect of Tridax procumbens in type 2 diabetes may be attributed to AMPK activation and suppression of hepatic gluconeogenesis (259.8)
Authors: Desai Gauri , Desai Shirish , Gavaskar Rajendra , Mathews Suresh
Published: June 2021

Traditional knowledge and anecdotal evidence suggest that Tridax procumbens Linn may have anti‐inflammatory and blood glucose lowering properties. Recent studies have shown that T.procumbens extract was effective in lowering blood glucose (42‐71% blood glucose reduction at 250‐500 mg/kg body weight) in alloxan‐induced diabetic rats. The goals of this study were to evaluate blood glucose lowering potential of T.procumbens extract in subjects with type 2 diabetes and characterize underlying molecular mechanisms. Twenty individuals with established type 2 diabetes were orally administered (15 ml, twice daily) T.procumbens extract, prepared according to Ayurveda guidelines, for a period of 4 weeks. Fasting blood glucose concentrations were reduced by 11% in men and 20% in women, and post‐prandial blood glucose concentrations were decreased by 26% in men and 29% in women. No adverse effects were reported by the participants. Chemical and microbial analyses of T. procumbens extract demonstrated the presence of phenolics, carotenoids, and flavonoids, and absence of microbial contaminants, aflatoxins, and heavy metals. T.procumbens extract demonstrated strong antioxidant potential and anti‐inflammatory activity as evidenced through Trolox equivalent antioxidant capacity, ferric‐ion reducing potential, hydrogen peroxide scavenging activity, and metal ion chelating effect in vitro, and inhibition of NFκB and IkBα in LPS‐stimulated RAW 264.7 murine macrophages. T.procumbens extract significantly increased AMPK activation and suppressed dexamethasone‐induced PEPCK and G6Pase gene expression with subsequent decrease in hepatic glucose production in H4IIE cells. Our studies indicate that T.procumbens extract exhibits strong antioxidant and anti‐inflammatory properties. Activation of AMPK and suppression of hepatic glucose production may be one mechanism that contributes to the glucose‐lowering property of T. procumbens.

Grant Funding Source: Diabetes Action Research and Education Foundation
The omega smart baby project: effect of maternal DHA on infant development (269.1)
Authors: Harris Mary , Stacy Miller , Baker Susan , McGirr Kathryn , Davalos Deana
Published: June 2021

A randomized placebo controlled trial was conducted to examine the effect of DHA on gestational length and cognitive development. 115 pregnant women were supplemented with 300 mg DHA/d or placebo during pregnancy and thru the first 3 mo of breastfeeding (BF). Bayley Scales of Infant Development (BSID) were administered at 4 mo and 1y. Dietary DHA intake during pregnancy, was 81.9 +/‐ 5.8 mg/day (95% CI 72.3,91.5) and increased only slightly during BF. Some took prenatal vitamins with 0– 300 mg DHA per day resulting in total DHA intakes of 80 to 1100 mg/day. RBC and plasma phospholipid DHA at entry (expressed as % total fatty acids) were5.94 +/‐ 1.52 and 5.04 +/‐ 1.44 (mean +/‐ SD), respectively. Breastmilk (BM) % total DHA was .53 +/‐ .037 (mean +/‐ sem) at 2 mo and decreased to .35 +/‐ /039 at 4 mo. BM DHA was significantly greater in the DHA treatment ( p<.02) and with the highest DHA intake compared to the lowest. BM DHA was lowest in carriers of the minor allele of the delta‐5 desaturase gene. Data were analyzed by treatment and by DHA intake. DHA supplementation resulted in a 1 wk increase in gestation with a significant (p<.03) 8 d increase in gestational length comparing the highest intake of DHA( > 600 mg/d) to the lowest (< 300 mg/d). BSID cognitive and language scores were 10 points higher with high intake of DHA compared to the lowest (p<.02). Maternal DHA impacts both pregnancy outcome and subsequent development.
Ellagic acid attenuates adipocyte and hepatic triglyceride contents via discrete mechanisms (269.6)
Authors: Okla Meshail , Kang Inhae , Lee Meeae , Chung Soonkyu
Published: June 2021

Liver and adipose tissues are major regulators of lipid metabolism. The aim of this study was to determine the anti‐lipogenic mechanisms of ellagic acid (EA), a naturally occurring polyphenol found in many berries and grapes, in adipocytes and hepatocytes. 3H‐olecic acid and 3H‐acetyl CoA were added to the primary cultures of human adipocytes and human hepatoma Huh7 cells to determine the effect of EA on fatty acid (FA) uptake, triglyceride (TG) esterification and TG de novo synthesis. In adipocytes, EA significantly reduced adipogenic gene expression and TG accumulation. Despite the substantial decrease of TG levels, EA had no significant impact on FA uptake or FA conversion into TG compared to the control group. However, the conversion of acetyl CoA into TG was dramatically reduced. In Huh7 cells, EA decreased TG accumulation in a dose‐dependent manner. Intriguingly, EA in liver cells did not only attenuate de novo synthesis of TG but also decreased FA uptake, and TG esterification. In addition, oxygen consumption was augmented by EA in Huh7 cells. Collectively, these data suggest that EA can attenuate TG contents in adipocytes as well as hepatocytes by altering distinct metabolic pathways.
The genetics and epigenetics of extreme healing: hypertrophic scars and diabetic ulcers (337.4)
Authors: Gibran Nicole
Published: June 2021

Hypertrophic scars and non‐healing diabetic ulcers represent wound complications on opposite ends of a continuum of healing. In spite of years of research, few breakthroughs in understanding the pathophysiology or developing therapeutic advances have relieved patient misery and health care costs for either chronic disease.

We combine in vitro endothelial studies and an in vivo diabetic murine model to elucidate epigenetic effects of a high fat Western diet on diabetic wound healing and cellular responses. We use in vitro fibroblast studies, a porcine model of fibroproliferative scarring and a translational clinical trial to correlate single nucleotide polymorphisms with long term scarring after burn injury. Together these approaches offer novel perspectives on cutaneous responses to injury.
Towards the genetic architecture of the human metabolome (350.1)
Authors: Gerszten Robert
Published: June 2021

In prior studies of the Framingham Heart Study (FHS) Offspring Cohort, we have identified and validated metabolite profiles of those destined to develop overt diabetes. The strongest individual predictors of future DM included branched chain amino acids, aromatic amino acids, specific triacylglyerol species, and 2‐aminoadipic acid (a putative lysine degradation product). These metabolites predict DM above and beyond clinical risk factors (such as body mass index, parental history) and biochemical markers (HOMA‐IR, OGTT). By integrating metabolite data with genome wide scans for common variants, we have identified 23 novel genetic determinants of human metabolism, including 8 loci previously implicated in human diseases. All of our data have been made publicly available to the broader scientific community.
Maternal determinants of pregnancy complications, birth outcomes, and postpartum depression (369.8)
Authors: Shin Dayeon , Kim WooKyoung , Song SuJin , Song Won O.
Published: June 2021

Various socio‐demographic and lifestyle factors have independently been reported to be associated with pregnancy complications, birth outcomes. We aimed to identify if prepregnancy BMI, gestational weight gain (GWG) and Adequacy of Prenatal Care Utilization (APNCU) are collective determinants of pregnancy complications, birth outcomes and postpartum depression. Multiple logistic regression analyses were conducted using the data of 7,911 Michigan’s 2007‐2010 Pregnancy Risk Assessment Monitoring System (PRAMS). After age, race/ethnicity, education, incomes, smoking status, and gestational weeks were controlled, prepregnancy BMI>30 was associated with high risk for large‐for gestational age (LGA) (aOR:1.97, 95% CI:1.61‐2.42), gestational hypertension (GH) (2.68, 2.23‐3.22), and gestational diabetes mellitus (GDM) (4.33, 2.79‐6.71) and low risk for small‐for‐gestational age (SGA) (0.70, 0.55‐0.87); excessive GWG increased the risk for LGA (1.73, 1.40‐2.12), macrosomia (3.23, 1.70‐6.13), and GH (1.36, 1.12‐1.64) and decreased the risk for SGA (0.56, 0.45‐0.68) and preterm labor (0.75, 0.64‐0.88). Compared to the women who started prenatal care in the first trimester, those who received no prenatal care had a higher risk for LGA (3.06, 1.05‐8.91). Prenatal care received during the 2nd or 3rd trimesters decreased the risk of having postpartum depression (0.73, 0.56‐0.95). However, APNCU was not associated with pregnancy complications, birth outcomes and postpartum depression. Maintaining healthy prepregnancy BMI and adequate GWG are important determinants for optimal health outcomes of pregnant women.
Different responses to soy and estradiol in the reproductive system of prepubertal male rats and neonatal male pigs (373.5)
Authors: Ronis Martin , Hennings Leah , Gomez‐Acevedo Horacio , Badger Thomas
Published: June 2021

Concerns have been raised regarding the safety of soy infant formula based on phytochemical components such as genistein, structurally similar to estradiol (E2). To examine potential estrogenic actions on male development, we fed weanling male rats casein‐based or soy protein isolate (SPI)‐based diets (PND21‐PND33) and neonatal male piglets cow’s milk or soy infant formulas (PND2‐PND21) or treated the animals with E2. E2 reduced testis and prostate weight and serum testosterone (P<0.05) in both species. Soy diet had no effects. In the piglet, E2 increased testis tubule area, Sertoli cell and germ cell number (P<0.05) and decreased expression of CYP19 and StAR mRNA. Soy increased germ cell number (P<0.05). The major effect of E2 is to change gene expression in reproductive tissues. E2 changed expression of 2645 genes in rat testis vs. 312 genes changed by SPI >1.5‐fold. 74% of the SPI genes were common to E2. In contrast, 353 genes were changed by E2 and only 116 by feeding soy formula in piglet testis with only 34 genes (29%) in common. These data suggest that soy diets are not estrogenic in the male reproductive system but may interact with a small subset of estrogen‐regulated genes in a species and developmental stage‐specific manner.

Grant Funding Source: USDA‐ARS CRIS 6251‐51999‐007‐04S
Milk glycan composition mediates gut microbiota, growth, and morbidity outcomes in Gambian infants (38.4)
Authors: Zivkovic Angela , Bernstein Robin , Huang Jincui , Totten Sarah , Lewis Zachery , Mills David , Prentice Andrew , Moore Sophie , Lebrilla Carlito
Published: June 2021

Human milk contains a variety of bioactive molecules that guide the development and growth of the newborn infant. Among them are various complex sugars or glycans, both as free oligosaccharides and as glycans bound to glycoproteins. Glycans in milk from mothers in the Gambia were examined in relation to infant gut microbiota, growth, and morbidity outcomes. Infants who were fast growers had higher relative concentrations of bifidobacteria, and were fed milks with higher % sialylated but not fucosylated oligosaccharides. On the other hand, the higher the % fucosylated oligosaccharides the less sick days the infant experienced. Growth rate was higher with increasing total oligosaccharide content of the milk. Higher levels of certain glycans on specific sites of two antibacterial proteins, lactoferrin and immunoglobulin A (IgA), were found in infants who had no sick days and low levels of fecal calprotectin, a measure of gut inflammation. In summary, glycosylation in human milk is an important mediator of gut microbiota composition, growth, and morbidity in developing infants.

Grant Funding Source: Supported by Gates Foundation Grand Challenges Explorations to R. Bernstein, #OPP1046163
Low‐income children feel safer walking/biking to school: physical activity‐related perceptions differ by income status in 5th graders (380.2)
Authors: Cardel Michelle , Gunnarsdottir Thrudur , Beck Jimikaye , Peters John , Hill James
Published: June 2021

Objectives: We examined baseline health‐related behaviors and perceptions by income status in children participating in 5th Gear Kids, a multi‐level childhood obesity prevention program in 5th Graders. Methods: Participants included a diverse group of children (41.44% White, 27.85% Hispanic, 16.77% African American, and 13.94% other) from two school districts in Colorado (n=6679; age 10.07). Anthropometrics were measured and health‐related behaviors and perceptions were assessed with a validated questionnaire (n=3023). Free/reduced lunch (FRL) prevalence in each school was used to group schools into income levels (HIGH <33% FRL; MED33‐66% FRL; and LOW >66% FRL). Results: Overweight/obesity was significantly more prevalent in LOW when compared to HIGH (35.8% vs. 13.0%). Over 90% of the total sample reported eating healthy was important to them. Though more LOW children reported they were more likely to pick the healthiest food option when it was made available to them, they consumed significantly more fast food and sugar‐sweetened beverages relative to HIGH. Significantly more HIGH children reported that physical activity (PA) was important to them and reported increased PA relative to LOW children. Nevertheless, HIGH children walked/biked to school significantly less than LOW and were significantly more likely to perceive walking/biking to school as dangerous. Conclusions: The importance of healthy eating did not differ between income groups; however, LOW children reported a higher prevalence of many dietary behaviors typically associated with obesity. Physical activity was rated higher in importance in the HIGH children, concurrent with significantly higher physical activity levels relative to LOW children. Interestingly, LOW children were more likely to bike/walk to school and were less likely to perceive walking/biking to school as dangerous. Differing perceptions across the socioeconomic gradient will be researched further in our longitudinal intervention.

Grant Funding Source: Colorado Health Foundation
The effects of whey protein supplementation on body composition and physical performance tests in men (381.2)
Authors: Bilgiç Pelin , Ersoy Gulgun , Ergun Nevin , Hongu Nobuko
Published: June 2021

The purpose of this study was to determine the effects of whey protein supplements on total and regional body compositions and physical performance tests in response to 10 weeks of resistance training. Twenty‐four men aged 18‐32 years, mean age 23 ± 3.4 years, were randomly divided into Protein (P) group, with whey protein supplementation (70 g/day on training days and 35 g/day on non‐training days) or Control (C) group, without supplementation. Daily mean total protein intakes of P and C groups were 118.2 ± 27.3 g (2.25 ± 0.26 g / kg BW) and 66.3 ± 15.0 g (1.16 ± 0.26g /kg BW), respectively (p < 0.001). All subjects trained 3 days/week for 10 weeks. At 0 and 10th weeks, subjects underwent bioelectrical impedance for total and regional body composition measurements and physical performance tests; i.e., one repetition maximum (1RM) strength, hand grip, and wall sit time tests. Data were analyzed using repeated measures ANOVA and presented as mean ±SD changes from baseline. No significant group x time interaction effects were observed among groups in changes in any variable. A significant time effect was observed for lean mass (P:2.3 ± 3.6 kg; C:0.8 ± 0.9 kg), mid‐upper arm area (P:8.0 ± 5.5 cm2; C:0.9 ± 2.5 cm2), bench press 1RM (P:18.5 ± 7.5 kg; C:11.4 ± 6.7 kg), wall sit time test (P:5.9 ± 18.4 sec; C:20.5 ± 31.2 sec), and handgrip test (P:4.9 ± 3.8; C:2.5 ± 2.2,) (all, p < 0.001). During resistance training, daily supplementation with whey protein was effective for promoting gains in lean mass and muscle strength.

Grant Funding Source: Supported by Hacettepe University Scientific Research Projects Coordination Unit
Consumption of an egg‐based breakfast reduces hunger and increases postprandial energy metabolism in normal weight and overweight school‐aged children (381.4)
Authors: Binns Ashley , Gray Michelle , Seo Han‐Seok , Zhang Baoyue , Luckett Curtis , Smith Keyona , Baum Jamie
Published: June 2021

In the US, one in every three children ages 2‐19 is overweight or obese. Although multifactorial in nature, obesity is primarily attributed to a mismatch between energy intake and energy expenditure (EE). One factor that influences this balance is thermic effect of food (TEF). Diets higher in protein have been shown to increase TEF. Breakfast is a key component of a healthy diet and can positively impact children’s health. It has been suggested that skipping breakfast may lead to greater hunger later in the day, resulting in greater overeating. The objective of this study was to determine if an egg/protein (PRO)‐based breakfast (22% PRO, 48% carbohydrate (CHO), 30% fat) would enhance TE, improve glycemic control and reduce hunger in NW (n=14) and OW (n=11) school‐aged children compared to an isocaloric CHO‐based breakfast (5% PRO, 65% CHO, 30% fat). Twenty‐three children (8‐12 y) completed both the PRO and CHO breakfast trials. EE was determined via indirect calorimetry at 0, 30, 60, 120, and 240 min postprandial (PP). Blood glucose (BG) was collected at 0, 15, 30, 60, 120, and 240 min PP and levels of hunger were assessed using a visual analog scale at same interval. At 240 min, an ad libitum buffet lunch was provided. Results indicate no significant difference in PP BG values between groups. However, PRO significantly reduced PP hunger (p<0.05) compared to CHO in NW and OW children. Consumption of PRO resulted in lower caloric intake at lunch (~100 kcal) compared to CHO. Relative to body weight, NW burn more calories per min compared to OW. NW have significantly higher TEF compared to OW (CHO p<0.001, PRO p=0.001). There was a trend (p=0.068) for higher TEF (10%) with PRO compared to CHO in OW. These data suggest that consumption of a PRO‐based breakfast has the potential to modulate PP energy metabolism in OW children through increased TEF, improved BG regulation, and decreased hunger.

Grant Funding Source: Supported by the American Egg Board/Egg Nutrition Center.
Low‐calorie sweeteners and body weight and composition: a meta‐analysis of randomized controlled trials and prospective cohorts (391.1)
Authors: Miller Paige , Perez Vanessa
Published: June 2021

Findings from randomized controlled trials (RCTs) and prospective cohorts investigating the relationship between low‐calorie sweeteners (LCS) and body weight and composition have been mixed and use of LCS as part of weight loss efforts remains controversial. The purpose of this study was to systematically review and quantitatively summarize results from RCTs and separately from prospective cohorts. Meta‐analyses evaluated weighted mean change in body weight and composition values between the LCS intervention and placebo groups among the 15 RCTs, and weighted mean correlations for LCS intake and body weight and composition parameters among the 9 prospective cohorts. Compared to placebo, LCS from food, beverages, or supplements significantly reduced body weight (‐0.74 kg, 95% CI: ‐1.10, ‐0.38), fat mass (‐1.18 kg, 95% CI: ‐1.75, ‐0.60), and waist circumference (‐0.93 cm, 95% CI: ‐1.74, ‐0.12), with no effect on BMI. In contrast, the meta‐analysis of cohorts showed that LCS intake was weakly but significantly associated with increased BMI (0.034, 95% CI: 0.009, 0.060), and nonsignificantly associated with increased body weight (0.024, 95% CI: ‐0.010, 0.058) and fat mass (0.035, 95% CI: ‐0.026, 0.096). Overall, available evidence from RCTs indicates that LCS may help reduce body weight, fat mass, and waist circumference, although this conclusion is not supported by results from prospective cohort studies.
Vitamin B12 supplementation during pregnancy and postpartum increases maternal plasma anti‐influenza IgA and ameliorates infant’s plasma AGP response (392.8)
Authors: Siddiqua Towfida , Ahsan Khalid , Ahmed Shaikh , Allen Lindsay , Raqib Rubhana
Published: June 2021

There is no information about effects of vitamin B12 supplementation during pregnancy and postpartum on immune function in mothers and their offspring. Women (n=66, 18‐35 y, Hb <115 g/L, 11‐14 wk pregnant) were randomized to receive 250 μg B12/d or a placebo through 4 mo postpartum. Both groups also received 400 μg folic acid + 60 mg Fe. Women were immunized with HINI vaccine at 26‐28 wk gestation. Vitamin B12, HINI‐specific IgG and IgA antibody responses in plasma, and IgA responses in colostrum/breast milk; acute phase response proteins, High sensitivity C‐reactive protein (HsCRP) and acid glycoprotein (AGP) were analyzed in plasma. At baseline, 26% of women were B12 deficient, and 39% had marginal status. Supplementation led to B12 adequate status in 94% mothers. In both groups combined, cord plasma H1N1 IgG was correlated with maternal (r=0.25, p=0.04) and infant (r=0.41, p=0.001) H1N1 IgG at 4 mo. Pre and post immunization H1N1‐specific maternal plasma IgG were similar in both groups. Supplementation significantly increased geometric mean titers of H1N1‐specific IgA in mothers at 4 mo postpartum compared to placebo group (p=0.03). Maternal supplementation reduced plasma AGP concentrations in infants at 4 mo compared to placebo group (0.75 vs. 0.62 g/L; p=0.05). Supplementation of 250 μg B12/d through pregnancy and lactation improved humoral immune function in response to antenatal influenza immunization, and reduced acute phase response.

Grant Funding Source: Nestle Foundation
Wnt/β‐catenin signaling in liver regeneration (398.5)
Authors: Yang Jing , Okabe Hirohisa , Williams Bart , Monga Satdarshan
Published: June 2021

β‐catenin plays a critical role in triggering liver regeneration (LR) after partial hepatectemy (PH). β‐catenin can be activated by numerous pathways, the primary one being Wnt signaling. Defect of β‐catenin leads to delay of LR in hepatocyte specific β‐catenin knockout mice (KO1). Ablation of Wnt‐dependent β‐catenin signaling in hepatocyte specific LRP5/6 double knockouts (KO2) also results in suboptimal LR after PH, which phenocopies KO1. Therefore, Wnt signaling is the major signaling pathway that activates β‐catenin during LR. To further elucidate the source of Wnts in the liver, we studied Wnts secretion from different hepatic cell population after PH. Wingless (Wls) is a critical regulator of Wnt secretion. We generated hepatocyte and macrophage specific Wls knockout mice (KO3&KO4, respectively), which lack the ability of hepatocytes and macrophages to secrete Wnts, respectively. KO3 had normal initiation of LR, while depletion of Wls from macrophages leads to suboptimal LR and impaired β‐catenin activation. Therefore, macrophages but not hepatocytes are responsible for Wnt secretion and β‐catenin activation during LR after PH.
Epigenetic regulation of the X chromosome (471.1)
Authors: Akhtar Asifa
Published: June 2021

Our lab is studying the chromatin and epigenetic mechanisms underlying X‐chromosome specific gene regulation using Drosophila dosage compensation as a model system. Dosage compensation mechanisms regulate the expression of X‐linked genes. In Drosophila this process leads to the transcriptional upregulation of the single male X chromosome and is regulated by a ribonucleoprotein male specific lethal (MSL) complex. The MSL complex localizes on hundreds of sites on the male X chromosome and is involved in hyperacetylation of histone H4 at lysine 16 by MOF, histone acetyltransferase. Interestingly, this complex is also conserved in mammals and appears to be involved in regulating diverse cellular processes. The recent progress of our work will be presented.
Estrogen deficiency induced intestinal inflammation and permeability is linked with osteoporosis (488.8)
Authors: Raehtz Sandra , Fedorko Alyssa , McCabe Laura
Published: June 2021

One out of two post‐menopausal women will experience an osteoporotic fracture. Osteoporosis is characterized by decreased bone density and increased fracture risk, over 2 million annually. We and others previously demonstrated an association between intestinal inflammation and bone loss. Given that menopause is associated with bone inflammation, we hypothesized that intestinal inflammation contributes to bone inflammation and loss associated with estrogen depletion. We performed ovariectomy surgeries to induce an estrogen deficient state in female mice. As expected, micro‐computed tomography demonstrated that distal femur trabecular bone volume was decreased by 26% as compared to sham control mice. Gene expression analysis of the small intestine demonstrated increased levels of pro‐inflammatory cytokines (tumor necrosis factor (TNF) and interferon gamma (IFNg)) and decreased anti‐inflammatory cytokines (interleukin‐10 (IL‐10)). Consistent with increased inflammation, estrogen depletion increased intestinal permeability. This was observed functionally as indicated by serum endotoxin levels that were increased by 29%. This was further demonstrated by decreased expression of key gap junction genes (claudin 15 (CLDN15) and junction adhesion molecule 3 (JAM3)). Pearson correlations demonstrate a positive association between intestinal inflammation and the extent of bone loss. Together, our studies indicate that intestinal inflammation is linked to bone loss. Understanding this association will identify new therapeutic targets to increase bone density and health.

Grant Funding Source: Michigan State University
Progressive digestion of limb‐specific Shh regulatory region reveals critical sequences necessary for enhancer activity (541.5)
Authors: Treto Jessica , Bailey Traci , Pira Charmaine , Oberg Kerby
Published: June 2021

Regulation of anterior‐posterior patterning of the limb bud during development is mediated by Sonic hedgehog (Shh), expressed in the zone of polarizing activity in the distal posterior region. Upstream of Shh lies a 1.8 kb limb‐specific Shh regulatory region (LSSRR) that regulates the expression of Shh. LSSRR can be subdivided into four highly conserved regions, A, B1, B2, and B3. Our goal is to determine which sequences within LSSRR are necessary for enhancer activity.

We progressively digested the LSSRR sequence from the 5’ end and the 3’ end. We generated expression constructs for each fragment containing the minimal HSV‐tk promoter linked to a green fluorescent protein (GFP) reporter. These constructs were electroporated into the chick presumptive limb bud. After 48 hrs, we visualized and recorded the GFP fluorescence to determine enhancer activity.

Removal of 217 3’ bases abolishes enhancer activity, while even after removal of 1161 bases from the 5’ end activity persists. Our progressive digestions indicate that enhancer activity is retained within 107 base pairs of the B3 conserved region.

This region contains three highly conserved motif clusters and is rich with potential binding sites including overlapping sites for retinoic acid receptor/NFB/ETS and sites for E‐Box and Blimp1.

Although other sequences may be required for inhibiting activity and chromatin looping, we conclude that this 107 base pair region is essential for LSSRR enhancer activity.

Grant Funding Source: Institutional
Epiploic bypass flap: a new method of limb salvage; anatomical bases and clinical application (547.9)
Authors: Settembre Nicla , Malikov Serguei , Branchereau Alain , Magnan Pierre‐Edouard , Braun Marc
Published: June 2021

Aim: The incidence of critical limb ischemia increases with the ageing of the population. Revascularization decreases the rate of amputation. The wounds infected with exposure of the tendons, bones or articulations cannot heal in spite of a bridging and local debridement. Surgery associating a distal venous bypass or recanalisation and a free flap makes it possible to treat the losses of substances and presents a hemodynamic advantage, by increasing the flow of the bypass, thanks to the vascular bed added by the flap. It is a complex surgery because of the multiplicity of anastomoses on the same arterial axis, with a risk of thromboses and complications related to the venous autograft. To mitigate these disadvantages, we proposed a new surgical method based on the use of a single anatomical unit, the epiploic bypass flap (BF), based on the gastroepiploic artery (GEA) as the inflow for a bypass and a free flap. The aim of this work was to analyze the anatomical feasibility of an epiploic bypass flap and to determine its limits.

Material and methods: 100 anatomical preparations were carried out with a measure of the internal and external diameters and the lengths of GEA and its branches as well as an X‐ray after injection of a radiopaque product. A first clinical application was carried out.

Results: our study confirm the anatomical feasibility of a bypass flap. The average available length of GEA is 245 mm (210‐280 mm). The average proximal diameter is 3 mm and the distal one is 1.5 mm. The most distal epiploic branch which feeds the bypass is approximately 180 mm (161‐195 mm) of the origin of the gastroepiploic artery. The anatomical unit based on the GEA provides an arterial graft relatively long and a large flap, malleable and resistant to infection.

Conclusions: Epiploic bypass flap is a surgical technique which allows distal revascularization and a simultaneous cover of the limb extremity. This technique can be useful in patients requiring a distal revascularization associated with a cutaneous cover.
Mechanisms of RNA polymerase II processing of DNA lesions formed by novel monofunctional platinum anticancer drugs (560.11)
Authors: Wang Dong
Published: June 2021

Pyriplatin and Phenanthriplatin, a novel class of platinum compounds, forms monofunctional DNA lesions having a structure and spectrum of anticancer activity distinct from those of the FDA‐approved drugs cisplatin, carboplatin, and oxaliplatin. These novel structure‐activity properties hold promise for overcoming drug resistance and improving the spectrum of treatable cancers over those responsive to cisplatin. However, the detailed molecular mechanism by which cells process DNA modified by pyriplatin and related monofunctional complexes is not at all understood. Here we report the structure of a transcribing RNA polymerase II (pol II) complex stalled at a site‐specific monofunctional pyriplatin‐DNA adduct in the active site. The results reveal a molecular mechanism of pol II transcription inhibition and drug action that is dramatically different from transcription inhibition by cisplatin and UV‐induced 1,2‐intrastrand cross‐links. In addition, we present the systematic mechanistic investigation that addresses how a site‐specific platinum‐DNA monofunctional adduct affects the Pol II elongation and transcriptional fidelity checkpoint steps, which is an important step toward elucidating the mechanism of action of these compounds. Our findings provide insight into structure‐activity relationships that may apply to the entire family of monofunctional DNA‐damaging agents and pave the way for rational improvement of monofunctional platinum anticancer drugs.
SRL lectin induced apoptosis of human colon cancer cells: identification of pathways leading to cell death (589.1)
Authors: Guha Nilanjan , SA Deepak , Barkeer Srikanth , KS Arun , Rhodes Jonathan , Yu Lu Gang , Swamy Bale M , Inamdar Shashikala
Published: June 2021

Sclerotiumrolfsii, a soil borne plant pathogenic fungus secrete a developmental‐stage specific lectin (SRL) which displays strong binding to TF antigen and its derivatives. Histochemical studies revealed specific binding to human cancerous colon tissues but not to normal ones. SRL exhibits strong receptor mediated antiproliferative activity towards human colon cancer cell lines by inducing apoptosis, involving the participation of caspase 8, 9 and 3. It has also been demonstrated that SRL at sub toxic concentrations completely inhibits tumour growth in NOD SCID mice models bearing HT29 xenografts when injected intratumorly. In this study, we investigated the signaling pathways that lead to apoptosis. HT29 cells were treated with lectin and samples were harvested at different time points and gene expression microarray analysis was performed using Agilent’s SurePrint G3 Human Gene Expression 8X60K V2 Microarray kit. Pathway analysis revealed that the MAPK signaling pathway is affected as early as 2h while cell cycle and the DNA damage response pathways are most significantly affected at later time points. Lectin treatment led to immediate over expression of the Jun transcription factor. miRNA expression study of the early time points showed very few miRNAs to be differentially regulated at 2 and 4 h after treatment. However,a significant change in differentially expressed miRNAs was noticed at 12h with the miRNA target list significantly overlapping with the differential gene expression list. The study suggests that the interaction of SRL with HT29 cells triggers apoptosis in these cancer cells by affecting the cell cycle pathway and concurrently inducing DNA damage. The observed effects are ,initiated through the activation of MAP kinase pathway and mediated by the transcription factor JUN. The findings will enable to exploit SRL for its diagnostic potential and also as a targeted drug for cancer therapeutics.
Comparative genomics of the response to cold shock in Saccharomyces paradoxus and Saccharomyces cerevisiae (589.3)
Authors: Dahlquist Kam , Harmon Nicolette , Amakiri Chidinma , Sherbina Katrina , Rohacz Nicholas , Fitzpatrick Ben
Published: June 2021

The growth rate of a wild species of yeast, Saccharomyces paradoxus, was compared to the domesticated species, Saccharomyces cerevisiae, when both were subjected to different temperatures and inhibitory drugs. We found that S. paradoxus had a shorter doubling time than S. cerevisiae at both 30 and 13°C, and that S. paradoxus grew significantly faster at the cold temperature than was expected by its warm temperature growth rate. Furthermore, S. paradoxus was more resistant to the growth‐inhibitory drugs, rapamycin and caffeine, than S. cerevisiae. To determine the gene expression changes responsible for resistance to cold shock, we performed a DNA microarray experiment on S. paradoxus. Cells were grown to early log phase at 30°C, then shifted to 13°C for 60 minutes, and then shifted back to 30°C for another 60 minutes. Samples were collected before cold shock (t0), after 15, 30, and 60 minutes of cold shock (t15, t30, t60), and after 30 and 60 minutes (t90, t120) of recovery at 30°C. Labeled aRNA from cold shock and recovery timepoints were paired with labeled aRNA from the t0 timepoint. Four independent replicates of the experiment were performed with dye swaps for two of the replicates. Cluster analysis shows two dominant patterns of expression: up‐regulation during cold shock followed by down‐regulation during recovery and vice‐versa. Genes involved in ribosome biogenesis show the former pattern of expression, similar to the pattern observed in S. cerevisiae. Further analysis is being performed to discern differences in expression between the two species.

Grant Funding Source: Supported by NSF RUI 0921038
Cadmium determinant 1 is a putative heavy‐metal transporter in Arabidopsis thaliana (617.4)
Authors: Aksoy Emre , Salazar Jose , Koiwa Hisashi
Published: June 2021

Cadmium (Cd) is a non‐essential toxic heavy metal for most organisms. Accumulation of Cd is toxic to plants, and it also causes serious health concerns in humans as plants are the major staples in human diet. In A. thaliana, cadmium is mainly accumulated in the roots suggesting that root‐to‐shoot Cd translocation is restricted at the xylem loading. Up to date, two members of HEAVY METAL P1B‐ATPase (HMA) family of transporters, namely HMA2 and HMA4, and some YSL transporters have been shown to be responsible for Cd loading into the xylem. Here we present data suggesting CADMIUM DETERMINANT 1 (CDM1) as a previously uncharacterized putative heavy‐metal transporter in plants. Expression of CDM1 was induced under Cd treatment in wild‐type seedling roots. Elemental analysis of two alleles of cdm1 mutant by ICP‐MS showed that the mutants accumulated less Cd and Zn both in shoots and roots after Cd treatment. In addition, root growth of cdm1 mutants showed higher tolerance to Cd toxicity. These data suggest that CADMIUM DETERMINANT 1 (CDM1) functions in Cd distribution in plants. Interestingly, preliminary confocal microscopy analyses of seedlings overexpressing CDM1:GFP indicated that CDM1 localized to the plastids. Further characterization of CDM1:GUS and CDM1 overexpression lines are in progress.

Grant Funding Source: Supported by NSF‐MCB 0950459
Impacts of erythropoietin combined with granulocyte‐colony stimulating factor on neural function after traumatic brain injury in rats (618.6)
Authors: Duan Hubin , Hao Chunyan , Li Shuzhen , Lu Gui , Fan Yimin
Published: June 2021

Objective: To investigate the effect of erythropoietin (EPO) and granulocyte‐colony stimulating factor (G‐CSF) combination therapy on functional recovery and nerve cell regeneration after TBI. Methods: The rats model with TBI was prepare by Feeney’s methods. 100 male Wistar rats were randomly divided into Sham operation group(Sham group), traumatic brain injury group (TBI group), EPO treatment group(T+E group ), G‐CSF treatment group(T+G group), and EPO combined G‐CSF group(T+E+G group), and then each group was sub‐divided into 5 subgroups according to the time 1,3 ,5,7 and 14 days after TBI.(Each subgroups n=4). Each treatment group rats were immediately given corresponding intervention measures after TBI for 3 days. Neurological function was assessed using a modified neurological severity score(mNSS) before rats were sacrificed at 1,3,5,7 and 14 days post TBI . Immunofluorescence measurement was used to detect newly proliferation cells number and its direction of differentiation by BrdU single positive cells and BrdU/NeuN or BrdU/GFAP double positive cells. Results: The mNSS scores of each treatment group was significantly lower than TBI group, and T+E+G group was also significantly lower than T+E group or T+G group at 5 ,7 and 14 days of surgery (P < 0.05); The BrdU positive cells number in T+E+G group was significantly higher than other groups starting from 3 days after TBI, and the peak at 7 days (P < 0.05). Compare the groups of brain tissue BrdU/NeuN and BrdU/GFAP double positive cells number, T+E+G group were significantly higher than other groups, T+E group and T+G group were significantly higher than Sham group and TBI group (P < 0.05). Conclusions: Combined use of EPO and G‐CSF can obviously improve functional recovery and promote the brain nerve cell regeneration differentiating to neurons and astrocytes in the injury boundary zone.

Grant Funding Source: This study was supported by the National Natural Science Foundation Youth (Fund No.30600637)
Human milk oligosaccharides: potential use in prevention of invasive candidiasis (623.6)
Authors: Gonia Sara , Tuepker Michele , Kim Jae , Bode Lars , Gale Cheryl
Published: June 2021

In premature infants, the opportunistic fungal pathogen Candida albicans is a leading cause of life‐threatening infections; the primary site for invasion being the intestine. The goal of this study was to test the hypothesis that Human Milk Oligosaccharides (HMOs) protect premature intestinal epithelial cells (IEC) from invasion by C. albicans. We used H4 cells, an established, primary, human fetal IEC line and measured fungal invasion immunohistochemically. The presence of HMOs, at concentrations present in human milk, resulted in an ~60% reduction in fungal invasion of H4 cells (p<0.05). Further, the HMO effect on invasion followed a dose‐response relationship. At slightly higher than physiologic concentrations, HMOs decreased the ability of C. albicans to make contact with H4 cells (p<0.05). In addition, at physiologic concentrations and above, HMOs reduced the overall lengths of C. albicans hyphae (p<0.05), the morphology associated with tissue invasion and injury. HMOs did not, however, reduce overall C. albicans growth rates in liquid or on solid growth media. In conclusion, HMOs decrease the ability of C. albicans to invade H4 cells, supporting a protective role for HMOs in the premature intestine. We speculate that this function of HMOs is attributed to their negative effect on C. albicans hyphal growth as well as blockade of physical interactions between host and fungal cells.