bluuebunny/crossref_metadata_2025 huggingface dataset:
Application of collagen coated silicone scaffolds for the three‐dimensional cell culture of primary rat hepatocytes
Authors: Stock Peggy , Winkelmann Claudia , Thonig Antje , Böttcher Gunther , Wenske Günther , Christ Bruno
Published: June 2021

Primary hepatocytes present an appropriate validation system for toxicological studies of novel agents in pharmaceutical‐ and cosmetics industries. Until today, three dimensional culture systems maintaining the functionality of hepatocytes are missing.

The aim was to analyse the applicability of 3D collagen‐coated silicone scaffolds versus 2D culture for culture of rat hepatocytes.

Primary rat hepatocytes were cultivated for 72h on multilayer lattice structures (A=23.7cm2) and compared with 2D culture on conventional dishes (A=8.9cm2). Cell counts, the growth pattern on silicone and the urea production rate were determined.

Based on identical initial cell counts the growth rate was about 42%±5% higher in 3D culture as compared to 2D. Under serum‐free conditions cells attached to the scaffold, formed clusters and organ‐like structures. The 3D culture on the silicone scaffold displayed no significant cytotoxicity after 72h: the percentages of dead cells were 7%±3% vs 9%±3%. The urea production rate [μg/105 cells] was significantly higher in the 3D vs 2D: 24h 9.1±2.7 vs 4.7±0.2; 48h 5.7±1.5 vs 3.7±0.05 72h 3.5±0.5 vs 2.1±0.1.

The results show the appropriateness of silicone scaffolds for the 3D culture of primary rat hepatocytes. Thus, the collagen‐coated silicone scaffolds represent an important tool for the development of 3D hepatocyte cultures sustaining the physiological functions.
Hsp70 and a novel axis of innate immunity in the virusinfected brain
Authors: Oglesbee Michael , Kim Mi Young , Shu Yaoling , Popovich Phil , Niewiesk Stefan
Published: June 2021

The major inducible 70 kDa heat shock protein (hsp70) promotes viral gene expression and stimulates innate immunity when released from cells, but it is unknown how virus‐hsp70 interaction influences outcome of infection. This deficiency was addressed using the mouse model of measles virus (MeV) neuronal infection. Transgenic mice were made to constitutively express hsp70 in neurons (a primate‐pattern of hsp70 expression) and were completely protected from MeV brain infection, compared to 35% mortality in non‐transgenic infected controls. Complete protection required a viral transcriptional response to hsp70. Transcriptome analysis of infected brain showed that hsp70‐dependent innate immunity was associated with an interferon beta (IFN‐β) response that was not observed in non‐transgenic mice. Disruption of the type 1 interferon receptor confirmed that hsp70‐dependent protection was IFN‐dependent. In vitro data established a model for the hsp70‐dependent innate immunity. We showed that MeV‐infection stimulates exosomal release of hsp70 from neuronal cells, and that extracellular hsp70 stimulates production of IFN‐β in uninfected brain macrophages (microglial cells) in a Toll‐like receptor‐dependent manner. Hsp70 can be released by numerous viral pathogens, supporting the potential broad relevance of this novel pathway of innate immunity. Studies were supported by R01NS31693.
Single incidence household food insecurity is associated with elevated maternal BMI
Authors: Tilton Nicholas Alfred , Black Maureen M , Magder Laurence S , Wang Yan , Hurley Kristen M
Published: June 2021

Objective

To examine the longitudinal relations among household food security status, maternal depressive symptoms and maternal BMI.

Methods

The Early Childhood Longitudinal Study–Birth Cohort (ECLS‐B) is a nationally representative sample of approximately 10,700 families, evaluated at 9m, 2, 4, and 5 yrs. Maternal BMI (measured), depressive symptoms (CES‐D 12‐item scale); and food security status (USDA 18‐item scale) were collected between 2001–2006. We applied a mixed effect model to examine weighted (adjusting for sampling design and non‐response) relations between baseline maternal depressive symptoms, household food security status (ever vs. never; across time points), and maternal BMI (yrs 2, 4, and 5).

Results

At baseline (9m), 13% of mothers reported moderate/severe depressive symptoms (CES‐D ≥12); 26% reported food insecurity at least once. Mean maternal BMI ranged from 22.9 at 9m to 23.8 at 5 yrs; 39% were overweight (BMI ≥ 25) by 5 yrs. In a model controlling for race/ethnicity and poverty level, mothers with ≥ 1 food insecurity report had a mean BMI 0.88 points higher at each time point than mothers with no food insecurity reports (p<0.001); mothers reporting baseline moderate/severe depressive symptoms had a marginally higher yearly BMI increase (0.08 BMI/yr, p=0.08) than mothers with mild/no symptoms.

Conclusion

Moderate/severe depressive symptoms at 9 m have a marginal effect on yearly maternal BMI increase. A single incidence of household food insecurity during early parenting is associated with higher maternal BMI.
Understanding Characteristics of Families Who Buy Local Produce
Authors: Racine Elizabeth F , Mumford Elizabeth A , Laditka Sarah B
Published: June 2021

Objective

To examine individual characteristics associated with local fresh produce purchasing among North Carolina families.

Design

Cross‐sectional analysis using data from the 2008 North Carolina Child Health Assessment and Monitoring Program (NC CHAMP), a representative sample of adults who have landline telephones.

Participants

Families with children who participated in the NC CHAMP (n= 2,932)

Main Outcome Measure

Frequency of buying local produce from local vendors in the past year

Analysis

Bivariate and multivariate zero‐inflated negative binomial regression identified characteristics associated with not buying local produce and, among those who purchased local produce, the frequency of purchasing.

Results

About half of families reported buying local produce on average once a month during the past year. In adjusted results, buying local produce was more likely among White families, lower income families, families living in rural areas, families with children who ate five or more servings of fruits and vegetables per day, and families with children in poorer health.

Conclusion and Implications

Findings suggest that it may be useful for dietitians and health promotion professionals to assess the availability of local produce in urban areas and to focus interventions to promote local produce purchases among Black families.

This study was not funded.
Children's lunch box after the implementation of a guideline to regulate the foods and beverages for sale in schools in Mexico City
Authors: Perez-Rodriguez Marcela , Naranjo Ijiel Barak , Pedraza Elizabeth , Ávalos Gabriela , Melendez Guillermo , Pfeffer Frania
Published: June 2021

Introduction

In January 2011 a guideline to regulate the foods and beverages (F&B) for sale in schools was implemented to encourage healthy foods consumption and to lower the sugar and fat in the F&B children consume.

Objective

Determine if the guideline is having an impact on the quality of the children's school lunch.

Methods

661 children from 1st to 3rd grade were interviewed; data on the F&B consumed the last 5 school days was recorded. Some anthropometric measurements were taken.

Results

Mean weight of children was 28.7±7.8 kg. 63.8%, 17.7% and 16.8% of children were normal weight, overweight and obese respectively. 6.5% bought their food at school, 37.2% brought theirs from home and 55.1% got it from either. 69.1% did not eat chips/fries and 13% consumed them =2 times. 22.9% consumed pastries/cookies =2 times and 56.1% did not eat any. Prepared foods (i.e. quesadillas) were consumed =2 times by 77.8% of children. 49.2% eat fruits and vegetables =2 times, but 20.9% did not consume them at all. 55.2% drank fruit beverages =2 times, all children consumed them at least once. 18.4% did not drink plain water and 73.7% drank water =2 times. 3 of every 4 kids bought water at school.

Conclusion

Most children bring food from home. Sugar sweetened beverages are highly consumed, plain water is poorly consumed. Special attention should be paid to foods prepared in cafeterias, due to being the most consumed but less regulated.
Diacylglycerol acyltransferase 1 (DGAT1) Functions as a Cellular “Hub” to Target Hepatitis C Virus Proteins NS5A and Core to Lipid Droplets
Authors: Ott Melanie , Camus Gregory , Herker Eva , Harris Charles , Farese Robert V.
Published: June 2021

The association of Hepatitits C Virus (HCV) proteins core and NS5A to lipid droplets plays an essential role in the HCV life cycle. Triglycerides stored in lipid droplets can be generated by two diacylglycerol acyltransferase enzymes, DGAT1 or DGAT2. We have previously shown that DGAT1 is critical for the recruitment of the HCV core protein to lipid droplets and thereby regulates viral assembly. This recruitment inhibits lipolysis of core‐coated droplets stabilizing these droplets as intracellular assembly platforms for HCV infection and potentially contributing to HCV‐induced steatosis. We screened all HCV proteins for interactions with DGAT1 and identified NS5A as a novel DGAT1 interaction partner. DGAT1, and not DGAT2, forms a trimolecular complex with core and NS5A and stabilizes the otherwise weak cellular interaction between the two viral proteins. Trafficking of NS5A to lipid droplets is dependent on active DGAT1 as shown in knockdown studies or after treatment with a specific DGAT1 inhibitor. Our data underscore the central role of DGAT1 as a host factor for HCV infection and identify DGAT1 as a novel link between core and NS5A proteins in cells. We propose that the newly identified interactions of DGAT1 with NS5A and core serve to guide trafficking of both viral proteins to the same subset of lipid droplets –those generated by DGAT1 ‐ where viral assembly can take place.
Vitamin D3 supplementation leads to increased total calcium absorption in 4 to 8 yr old children
Authors: Abrams Steven , Hawthorne Keli , Chen zhensheng
Published: June 2021

Few vitamin D supplementation trials exist in small children. We randomized 63 children, 4 – 8 yrs of age to 1000 IU/d vitamin D3 or placebo for 8 weeks. Baseline vitamin D intake was 221(79) IU/d and calcium intake was 830(197) mg/d. Baseline 25‐OHD was 27.7(7.2) ng/mL, mean(SD). Baseline 25‐OHD was not significantly correlated to fractional (alpha) or total calcium absorption (Va). BMI Z‐score was positively correlated to 25‐ OHD (r = 0.33, p = 0.009). After 8 weeks, using 25‐OHD as a covariate, Va was greater in supplemented vs non‐supplemented children, 281(18) vs 224(18) mg/d, mean(SEM), p = 0.033. When adjusted for calcium intake, there was a trend for a greater alpha with supplementation, 35.6(2.0) vs 30.7(2.0)%, p = 0.098 mean(SEM). Serum 25‐OHD and 1,25‐(OH)2D were not significant determinants of Va or alpha. We conclude that 1000 IU/d vitamin D3 supplementation increases calcium absorption in children with vitamin D intakes below recommended amounts. Optimal vitamin D intake and the relationship between serum 25‐ OHD and calcium absorption in small children requires further evaluation.
A complete meal based algorithm for predicting nonheme iron absorption
Authors: Armah Seth Mensah , Carriquiry Alicia , Cook James D , Reddy Manju B.
Published: June 2021

It is not feasible to conduct human studies to study all possible combinations of the various factors that affect nonheme iron absorption. Many algorithms have been developed in the past to estimate nonheme iron absorption based on single meal studies. However, exaggerating effect of the factors in single meal studies is well known. The objective of this study was to develop an algorithm based on a complete diet for estimating nonheme iron absorption. We used data from 4 complete diet studies that had 12–14 subjects who consumed 3 different diets (n=159), typical, high and low meat, tea, calcium and vitamin C. Multiple linear regression with a mixed effect model was used. Our results suggest that serum ferritin contributed significantly to the variability in nonheme iron absorption and the effect of dietary factors was small. The predicted absorption values correlated significantly with the observed values from single and complete meal studies (R2=0.57, P<0.001 and R2= 0.93, P< 0.0001, respectively). When iron absorption was predicted from intervention study data in subjects with ≤15μg/L serum ferritin, the values were similar to observed values (18.6 vs 18.8%). Although interpersonal variation explains a large proportion of the differences in nonheme iron absorption, our algorithms with complete meals will be useful in predicting nonheme iron absorption from usual diets of varied populations.

Grant Funding Source : IFPRI/HarvestPLUS and MI
Effects of weight loss or exercise on muscle quality in obese older women
Authors: Evans Ellen M , Larson Rebecca D , Johnson Curtis L , Guest Dolores D , Das Bhibha M , Ward Christie L , Chen Danchin D , Georgiadis John G
Published: June 2021

A key component in lower extremity physical function in older obese women is leg muscle quality (MQ), defined as: a) strength per muscle quantity or b) the degree of adipose intermuscular adipose tissue (IMAT) within the muscle. This pilot study assessed the relative impact of a weight loss diet (WL‐D) or exercise (EX) on MQ. Sedentary obese (BMI ≥ 30–35 kg/m2) women (N = 30) were randomized to a WL‐D or EX intervention for 16 weeks. Thigh subcutaneous adipose tissue (SAT), IMAT, and muscle (MUSC) was quantified by MRI. Strength was assessed via maximal isometric (IM) and isokinetic (at 60 deg/sec, IK60) contractions using an isokinetic dynamometer and calculated as Nm/(m3). WL‐D (n=8, 65.6±8.0 y) lost 2.4±3.9 kg of body weight (p=0.12) and EX (n=11, 63.7±4.5 y) lost did not change (0.06±1.8 kg; p=0.91). Changes in upper leg composition or MQ measures did not significantly differ between groups; however; a) SAT decreased 3.5% in WL‐D (p=0.11) and 3% in EX (p=0.01), b) IMAT increased 7% in WL‐D (p=0.04) but only 2.7% in EX (p=0.27), c) MUSC increased 3.6% in WL‐D (p=0.34) and 4.5% in EX (p=0.06), d) EX increased MQ‐IM 7.3% (p=0.17) and MQ‐IK60 1.6% (p=0.80) whereas WL‐D increased MQ‐IM 0.89% (p=0.79) and decreased MQ‐IK60 −8.2% (p=0.12). Weight loss and exercise may have differential effects on thigh composition and MQ in older obese women. Supported by NIH‐HL090455
Chronic antibiotic administration increases intestinal mass but inhibits functional adaptation in an intestinal failure piglet model
Authors: Barnes Jennifer L. , Tappenden Kelly A.
Published: June 2021

Antibiotic therapy is frequently used in pediatric intestinal failure (IF) patients but the effects on intestinal adaptation are unknown. Neonatal piglets underwent an 80% jejunoileal resection (n=39) and received 80% of nutrient needs parenterally and 20% enterally. The antibiotic NAXEL® was administered for 48 hrs or 7 days and piglets were euthanized 7 days after surgery. Variables of adaptation were assessed, analyzed as a randomized complete block design and considered significant at p≤0.05. The objective was to determine the effect of chronic antibiotic administration on intestinal adaptation in a model of neonatal IF.

Long‐term antibiotic use increased intestinal weight evidenced by an increase in ileal and colonic mucosa (mg/cm, p=0.01, 0.004 respectively) and total intestinal mass (g, p=0.03). Protein decreased compared to 48 hrs of antibiotics in all segments (μg/mg mucosa, p=0.02 duodenum, p<0.0001 jejunum, ileum and colon) and protein per cell (μg protein/μg DNA) decreased (p=0.02 duodenum, p=0.003 jejunum). Functional adaptation measured by modified Ussing chambers indicated a decrease in active ion (p=0.006 jejunum, 0.03 colon) and total ion transport (p=0.004 jejunum, p=0.05 ileum) after 7 days of antibiotic compared to 48 hrs. These results suggest that chronic antibiotic administration induces an increase in intestinal mass, but hinders functional adaptation in a model of neonatal IF.

Funded by NIH Ruth L. Kirschstein Institutional National Research Service Award 5T32 DK59802.
Adaptation to every other day feeding leads to an increase in small intestine transit‐amplifying cells and a decrease in gonadal fat
Authors: Mah Amanda T , Van Landeghem Laurianne , Blue R. Eric , Lund P. Kay
Published: June 2021

Caloric restriction delays aging. An every other day diet (EOD) involves food ad libitum (AL) for 24 hours then food withdrawal for 24 hours. EOD results in mild or no overall caloric restriction but can improve insulin signaling and longevity. We hypothesized that EOD would impact intestinal epithelial renewal. Sox9‐EGFP reporter mice express different levels of the Sox9‐EGFP transgene in intestinal epithelial stem cells (IESC; Sox9‐EGFP Low), transit‐amplifying progenitors (TA; Sox9‐EGFP Sublow), enteroendocrine cells (EEC; Sox9‐EGFP High), and other differentiated intestinal epithelial cells (IEC; Sox9‐EGFP Negative). Sox9‐EGFP mice were fed AL or EOD for 20 weeks. Body weight, small intestine and colon weight and length did not differ between diets. EOD mice had significantly lower gonadal fat mass (60%) and no detectable mesenteric fat. Flow cytometry revealed a two‐fold increase in the proportion of jejunal Sox9‐EGFP Sublow TA progenitors in EOD mice. Cyclic food deprivation decreases adiposity without affecting body weight. Selective increases in TA progenitors may reflect an adaptation to the EOD that maximizes the proliferative capacity of the intestinal epithelium to rapidly provide absorptive villus epithelial cells when fed. Taken together, this research begins to characterize the effect of the EOD paradigm on the intestine and body composition. Grants: NIH DK040247‐19, NIH 2‐T32‐DK07686

Grant Funding Source : NIH DK040247‐19
Making sense of dietary supplement research and a framework for the future
Authors: Coates Paul M , Bailey Regan L
Published: June 2021

Dietary supplements are widely used by Americans (53% of adults; 35% of children), with an annual sales volume of about $28 billion in 2009. Research on the health effects of dietary supplements is equivocal at best and the data in the scientific literature are subject to interpretation. In order to inform the scientific and consumer communities, the Office of Dietary Supplements has identified several gaps that need to be addressed, and has sought to develop tools to meet those needs: (1) systematic review of the relevant literature; (2) development of analytical tools to ensure accurate measurement of the content of dietary supplements; (3) understanding the appropriate methods to assess total nutrient intakes from both supplements and foods; and, (4) accurate measurement of biomarkers of nutrient status. Armed with this type of information the scientific community can develop a research agenda, hopefully one that that can be effectively translated to educate consumers and their healthcare providers and inform policy decisions. This talk will address the strengths and limitations of the available data on dietary supplements in chronic disease and discuss strategies to advance this field of research.
Dietary supplement users report physicians are primary source of information in East and West geographic regions
Authors: Kazaks Alexandra , Rozga Mary , Stern Judith
Published: June 2021

Objective

To assess primary sources of information about dietary supplements (DS) and to determine if the sources differ between geographic regions.

Methods

Questionnaires and verification of supplement labels were used to assess DS use and sources of information for 646 healthy male and female subjects aged 21–75 who had been recruited for lipoprotein assessment studies at Tufts University and UC Davis. A chi‐square test with an alpha of 0.05 was used.

Results

DS were taken by 65.2% of participants (M 49.5%; F 50.5%). In contrast to earlier studies that showed physicians were infrequent sources of DS information, this study found the most common source of DS advice for all respondents was physicians (52.4%). Other sources of information differed significantly depending on geographic location. Those in the east were more likely to ask health food retailers or pharmacists (p<0.001). Those in the west were more likely to rely on family (p=0.002) and friends (p=0.021) and magazines/journals/news and the internet (p<0.001).

Conclusion

When DS use is discussed with health care providers more informed use and fewer possible drug‐DS interactions may occur. Physicians may benefit from increased education about DS since they will be accountable for accurate information regarding DS use. (Research support: Bastyr University)
The Effect of Eating Speed on Energy Intake in Normal Weight and Overweight/Obese Subjects
Authors: Copeland Jennifer , James Ashlei , Chatley Brittany , Brown Kate , Pelzel Kristin , Crane Jennifer , Mashni Samar , Dart Lyn , Rhea Debbie , Shah Meena
Published: June 2021

The effect of eating speed on energy intake is controversial and whether it is affected by weight status in not clear. This was further examined by recruiting 35 normal weight (NW) (23.9±2.6 kg/m2) and 35 overweight/obese (OW/O) (31.2 ±4.6 kg/m2) subjects. Subjects came to a metabolic laboratory on two different days, separated by 13.8±12.7 days, for lunch. During the first day, each subject was randomly assigned to eat at either a fast or slow eating speed. The subject was then given a bowl of vegetarian pasta (women: 900 g; men: 1200 g) and instructed to eat as much as desired but at the assigned eating speed. During the second study day, the subject consumed the same meal but at the alternate eating speed. The amount of energy consumed was calculated. A mixed effects ANOVA model was used to examine the effect of eating speed and eating speed by weight status on energy intake. Energy intake was significantly lower at the slow compared to the fast eating speed (733.5±379.9 kcal vs. 806.2±349.8, respectively; p=0.018). Energy intakes at the slow and fast eating speeds were 801.9±437.5 kcal and 890.0±329.4 kcal, respectively, in NW subjects and 655.1±303.1 kcal and 722.5±354.3 kcal respectively, in OW/O subjects. There was no eating speed by weight status interaction effect on energy intake (p=0.62). Eating speed had a significant impact on energy intake but the response was not affected by weight status. This study was funded by TCU.
A mixture of grape seed and skin extract, green tea extract, resveratrol, and quercetin reduces blood pressure in hypertensive subjects with metabolic syndrome
Authors: Jalili Thunder , Beisinger Shara , Quadros Ashley , Rabovsky Alexander
Published: June 2021

We hypothesized that a supplement combining extracts from grape seed and skin (330 mg), green tea extract (100 mg), resveratrol (60 mg), and quercetin (50 mg) would reduce blood pressure (BP) in subjects with metabolic syndrome that had systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg. Individuals (n=20) meeting study criteria enrolled in a double blinded, placebo controlled, cross over trial. The 4‐wk placebo and supplement phases were separated by a 2‐week washout period. There was a −4.7 and −4.8 mmHg (p<0.05) reduction in 12h ambulatory diastolic and mean arterial BP, respectively, after supplementation vs. placebo. Though systolic BP was unchanged, higher baseline BP was significantly (p<0.05) correlated (R2=0.56) with larger reductions in systolic BP. Plasma angiotensin converting enzyme (ACE) activity and concentrations of urine nitrate, nitrite, and isoprostanes were measured to determine the mechanism of BP reduction. ACE activity and urine isoprostanes were unchanged, but a trend existed (p=0.098) toward greater urinary nitrate/nitrite after the supplement phase. Markers of inflammation (IL8, IP‐10, MCP‐1, CRP, TIMP‐1) were not altered. We conclude that this combination of polyphenolics can lower BP in humans with Stage 1 hypertension. We speculate the mechanism may be due to enhanced nitric oxide production. Funded by a grant from Melaluca, Inc.
Effect of fructose on triglycerides: a meta‐analysis of controlled feeding trials
Authors: Chiavaroli Laura , Sievenpiper John L , Mirrahimi Arash , Cozma Adrian I , de Souza Russell J , Yu Matthew , Carleton Amanda J , Beyene Joseph , Kendall Cyril WC , Jenkins David JA
Published: June 2021

Background

Health agencies have expressed concern that fructose may contribute to hyper‐triglyceridemia (HTG) in people with and without diabetes (DM).

Purpose

To investigate the effect of fructose on triglycerides (TG), we conducted a meta‐analysis of controlled feeding trials.

Methods

We searched MEDLINE, EMBASE, CINAHL and the Cochrane Library for relevant trials of ≥7days. Data were aggregated by the generic inverse variance method using random effects models and expressed as mean differences (MD) with 95% confidence intervals (CI). Heterogeneity was assessed (Chi2) and quantified (I2). Study quality was assessed by the Heyland score.

Results

46 isocaloric (n=549 non‐DM, n=174 DM) and 7 hypercaloric (n=127 non‐DM) trials met eligibility criteria. In isocaloric trials, fructose did not significantly effect TG (MD= 0.05 [95% CI: −0.02, 0.12]) with significant evidence of inter‐study heterogeneity. There was no effect modification by diabetes status. In hypercaloric feeding trials, fructose had a TG‐raising effect (MD=0.32 [95% CI: 0.08, 0.56]) with significant inter‐study heterogeneity.

Limitations

The majority of the trials were <12‐weeks and of poor study quality.

Conclusions

Isocaloric fructose does not have significant TG‐raising effects whereas hypercaloric trials did, however excess energy may be a cofounder.

Funding: Canadian Institutes of Health Research (CIHR).

Grant Funding Source : ASN
Alcohol and breast cancer risk in five ethnic groups: the Multiethnic Cohort
Authors: Park Song-Yi , Kolonel Laurence N. , Henderson Brian E. , Wilkens Lynne R.
Published: June 2021

To assess the relationship of alcohol consumption with postmenopausal breast cancer risk in a multiethnic population, we prospectively examined the association in 85,089 women enrolled in the Multiethnic Cohort in Hawaii and California. During an average follow‐up of 12.4 years, we identified 3,885 incident invasive cases. Hazard ratios (HR) and 95% confidence intervals (95% CI) were estimated using Cox proportional hazard models, controlling for potential confounders. Higher alcohol consumption was associated with increased risk of breast cancer: compared to nondrinkers, HRs were 1.23 (95% CI: 1.06, 1.42), 1.21 (95% CI: 1.01, 1.45), 1.12 (95% CI: 0.95, 1.31), and 1.53 (95% CI: 1.32, 1.77) for 5‐<10, 10‐<15, 15‐<30, and ≥30 g/day of alcohol, respectively. The positive association was seen in African American, Japanese American, Latino, white, but not Native Hawaiian, women, and in those with tumors that were ER+/PR+, ER+/PR‐, and ER‐/PR‐. The findings from this prospective study support the hypothesis that moderate alcohol consumption increases breast cancer risk in four ethnic groups, independent of ER/PR status. This work was supported in part by the National Cancer Institute (grant number R37 CA54281).
Living WCRF Recommendations associated with less Prostate Cancer Aggressiveness among African and Caucasian Americans
Authors: Arab Lenore , Su Joseph , Steck Susan , Ang Alfonso , Fontham Elizabeth , Bensen Jeannette , Mohler James
Published: June 2021

The relationship between adherence to World Cancer Research Fund recommendations and prostate cancer aggressiveness was examined using data from the population‐based North Carolina‐ Louisiana Prostate Cancer Project. Clinical criteria were used to classify prostate cancer aggressiveness at diagnosis for 1,049 African‐American (AA) and 1,083 Caucasian‐American (CA) participants. An association between prostate cancer aggressiveness and adherence to guidelines, measured on a scale of 0–9 was assessed by examining odds ratios (OR) and 95% confidence intervals (CI) adjusted for potential confounders. A significantly inverse association was found between prostate cancer aggressiveness and adherence with each additional point associated with a 13% reduction in risk of aggressiveness. A 38% increase in likelihood of aggressiveness was associated with adherence to fewer than 4 recommendations (OR 1.38, 95%CI=1.10–1.74) as compared to those who adhered to 4 or more recommendations. Results stratified by race suggested the association is equally strong in AA as among CAs (ORs 1.36 95%CI= 1.01–1.85; 1.41 95% CI= 1.01–1.98 respectively). This study shows that recommendations aimed at preventing cancer may also be associated with the occurrence of less aggressive prostate cancer in those destined to develop the disease.
Intestinal permeability at baseline is negatively associated with change in plasma zinc concentration following short‐term zinc supplementation
Authors: Wessells K. Ryan , Hess Sonja Y. , Ouédraogo Zinewendé P. , Rouamba Noel , Goto Rie , Ouédraogo Jean-Bosco , Brown Kenneth H.
Published: June 2021

Subclinical enteropathy is associated with malabsorption of fats, carbohydrates, and vitamins A, B12 and folate; however, little information is available on enteropathies and mineral absorption. Therefore, we assessed the relationship between intestinal mucosal function, as measured by the lactulose:mannitol (L:M) permeability test, and zinc (Zn) absorption, as estimated by the change in plasma zinc concentration (PZC) following short‐term Zn or placebo supplementation during a randomized community trial among 284 asymptomatic children 6–23 mo in Burkina Faso. Participants received either 5 mg Zn as zinc sulfate, or placebo, daily for 21 d. Mean ± SD PZC (μg/dL) at baseline was 62.9 ± 11.9 μg/dL and 63.6% of children had PZC < 65 μg/dL. Geometric mean ± SEM L:M recovery ratio was 0.05 ± 0.002 and the prevalence of altered intestinal permeability (L:M recovery ratio ≥ 0.07) was 25.7%. Following the supplementation period, the mean ± SD change in PZC (μg/dL) was significantly greater in the Zn supplemented group (15.6 ± 13.4 μg/dL) compared to the placebo group (−0.1 ± 10.9 μg/dL; P < .0001, ANOVA). Change in PZC was negatively associated with the L:M recovery ratio (P = 0.03), regardless of study group (P = 0.70). Short‐term Zn supplementation results in a large increase in PZC, but the magnitude of this response is affected by the presence of subclinical enteropathy as diagnosed by altered intestinal permeability.

Grant Funding Source: Nutsiset, SAS (Malaunay, France); Henry A. Jastro Graduate Research Scholarship Award (UC Davis); Bristol‐Myers Squibb Freedom to Discover Award
Maternal Iodine Deficiency during Pregnancy and Child Growth to 5 Years of Age in Rural Bangladesh
Authors: Jing Hua , Shamim Abu Ahmed , Schulze Kerry J , Jahan Nusrat , Ali Hasmot , Labrique Alain , Christian Parul , West Keith P
Published: June 2021

We examined associations between maternal gestational iodine status and growth to 5 years of age in 450 children in Gaibandha District, Bangladesh. Iodine status of women participating in a trial of vitamin A and beta‐carotene supplementation was assessed by urinary iodine excretion (UIE) early (<3rd trimester) and late (3rd trimester) in gestation. Growth variables at birth and ~60 months of age were weight (kg), length/height (cm), head circumference (HC, cm), their Z‐scores (WAZ, LAZ, HCAZ), and chest (CC, cm) and mid‐upper arm (MUAC, cm) circumferences. In multiple linear regression analyses, adjusting for confounders, children born to mothers with severe ID (UIE < 20μg/L) in late pregnancy were lighter in weight (β=−0.156 kg) and WAZ (β=−0.40), shorter in length (β=−0.81 cm) and LAZ (β=−0.43), and smaller in CC (β= −0.74 cm) at 5 years than children of mothers with adequate status (>100μg/L, p<0.05). UIE in early, but not late, pregnancy was positively associated with postnatal 5 year increments in weight (β=0.0023), WAZ (β=0.0016), height (β=0.0051), HAZ (β=0.0021) and HC (β=0.0018) (all p<0.05). Maternal iodine status, measured by gestational UIE, and reflecting iodine availability to the fetus, is a determinant of child growth over the first five years of life. Support: Gates Foundation, DSM Scholarship for International Micronutrient Research, USAID, Sight and Life Research Institute
Glypican‐3 as a Potential Therapeutic Target for Hepatocellular Carcinoma Immunotherapy
Authors: Trinh Thu , Liu Chen
Published: June 2021

Objectives

Glypican‐3 (GPC3), a cell surface heparin sulfate proteoglycan, has been shown to be highly expressed in hepatocellular carcinoma (HCC). The objective of our study is to examine the GPC3 protein processing in different HCC cell lines and human HCC tissues and to identify HCC‐specific target for immunotherapy.

Methods

Normal primary hepatocytes, 5 different HCC cell lines and 40 pairs of human tumor (T) and non‐tumor (N) liver tissues (each pair from the same patient) were subjected for detection of GPC3 by Western blot. Two HCC cell lines LH86 and Huh7 were incubated with different commercial anti‐GPC3 antibodies and cell viability was determined by MTT assay.

Results

Out of 40 pairs, we found in 27 pairs (67.5%) GPC3 was processed only in tumor part. 4 over 5 HCC cell lines (80%) also expressed cleavage pattern of GPC3, whereas normal hepatocytes are negative for GPC3 expression. Anti‐GPC3 antibody specific for C‐terminal inhibited cell proliferation in dose‐dependent manner, with LH86 more susceptible to C‐17 mAb than Huh7.

Conclusion

We concluded that C‐terminal of GPC3 is up‐regulated in HCC. Antibodies against GPC3 inhibited cancer cells proliferation through interaction with C‐terminal domain of GPC3, implying that this portion of GPC3 can serve as novel therapeutic target for HCC immunotherapy.

This work is supported by NIH grants R01CA133086 and K26 RR023976 to CL
Lean Growth Is Enhanced by Intermittent Bolus Compared with Continuous Feeding in Neonates
Authors: El-Kadi Samer Wassim , Boutry Claire , Gazzaneo Maria Carolina , Suryawan Agus , Orellana Renán , Srivastava Neeraj , Nguyen Hanh V. , Fiorotto Marta L. , Davis Teresa A.
Published: June 2021

Orogastric tube feeding is indicated in neonates with impaired ability to ingest food normally, and can be administered by intermittent bolus (INT) or continuous (CON) infusion. The aim of this study was to determine if these feeding modalities affect growth and lean tissue accretion. Neonatal pigs (n=6, 6‐d‐old) were fed the same diet in equivalent amounts continuously or intermittently (meal every 4h) for 21d. Weight gain was greater for INT than for CON pigs and resulted in heavier body weights from 9 until 21d of feeding (P<0.05). Percent body fat and lean body mass, determined by dual‐energy X‐ray absorptiometry (DXA), were not affected by feeding modality. However, lean tissue mass and spine length were greater in INT than CON pigs (P<0.05). Glucose and arterial insulin levels measured on the last day of feeding were greater for INT after the meal than for CON pigs (P<0.05). Muscle protein synthesis increased by 40 % in longissimus dorsi, and 34 % in gastrocnemius and soleus of the INT as compared to the CON group (P<0.05). Longissimus dorsi, semitendinosus, and soleus muscles were 56, 42, and 35 % heavier for pigs in the INT as compared to the CON group (P<0.05). These results suggest that intermittent feeding enhances lean tissue accretion as compared to continuous feeding, and that this difference is apparent by 9d and persists for the duration of feeding. (Supported by NIH AR444474 and USDA/ARS 6250‐51000‐055)
Threonine treatment increases heat shock protein 25 and 70 expression and decreases apoptosis in heat stressed intestinal epithelial cells
Authors: Baird Christine Hamiel , Kallweit Alyssa Rene , Beck Ryan , Niederlechner Stefanie , Wischmeyer Paul Edmund
Published: June 2021

Osmotically acting amino acids can be cytoprotective following injury in vitro and in vivo. Heat shock proteins (HSPs) have been implicated as one mechanism of protection in these models. Threonine (THR) is necessary for mucin production and gut function but it's effects on HSPs are unknown. Our purpose was to investigate if THR can protect Intestinal Epithelial‐18 cells (IEC‐18) from lethal heat stress (HS) and to elucidate it's mechanism of protection. Cells were treated for 15 min +/− THR and subsequent HS injury. Cell size was evaluated via microscopy. Survival was evaluated via MTS assay and cleaved caspase‐3 (CC3). Western blot analysis was used for CC3, HSP25, and HSP70 levels. THR increased survival in a dose dependent manner (p<0.02 vs. HS alone). THR increased cell size by more than 40% (p<0.001 vs. non‐THR groups). THR decreased CC3 more than 54%, and increased HSP25 and HSP70 levels more than 5 fold in HS (p<0.02 vs. HS alone). This is the first demonstration that THR can decrease apoptosis in heat stressed intestinal cells. THR induces cell swelling and it's effects on HSP25 and HSP70 may be an integral component of THR's mechanism of cellular protection.

Grant Funding Source : NIH
Calcium plus vitamin D supplementation during pregnancy protects Brazilian adolescent mothers from excessive bone loss during lactation
Authors: Diogenes Maria Eduarda L. , Bezerra Flávia F. , Rezende Eliane P. , Taveira Maria Fernanda , Pinhal Isabel , Donangelo Carmen M.
Published: June 2021

We evaluated the influence of calcium plus vitamin D supplementation during pregnancy on bone mass of lactating adolescent mothers (13–19y) with habitually low calcium (655 mg/d) and vitamin D (34 IU/d) intake. Total body (TB) and lumbar spine (L1‐L4, LS) bone mineral content (BMC) and density (BMD) were assessed by DXA at 1 and 4 mo postpartum (PP). Adolescent mothers randomly received daily calcium (600 mg) plus vitamin D3 (200 IU) supplement (n=30) or placebo (n=25) from 26 wk of gestation until parturition. Comparisons between groups were done using maternal age, height, season and period PP as covariates (ANOVA). At 1 mo PP, supplemented mothers had higher TBBMD (3%, P=0.05), LSBMD (4%, P=0.08), TBBMC (2.4%, P=0.06), and LSBMC (8.5%, P<0.05) than those in the placebo group. At 4 mo PP, differences between groups were more pronounced, with higher TBBMD (4.3%, P<0.05), LSBMD (5.9%, P<0.05), TBBMC (4.1%, P=0.06) and LSBMC (9.1%, P<0.05) in the supplemented group. Our results indicate that calcium plus vitamin D3 supplementation during pregnancy protects adolescent mothers from excessive bone loss during lactation, especially at the lumbar spine. This protective effect persists at least up to 4 months of lactation.

Financial support: CNPq and FAPERJ (Brazil).
A 3‐D geometric morphometric study of intraspecific variation in the ontogeny of the temporal bone in modern Homo sapiens
Authors: Smith Heather F , Ritzman Terrence , Otárola-Castillo Erik
Published: June 2021

The morphology of the temporal bone has been determined to reflect both phylogenetic relationships among hominoid species and population history among modern human groups. It is a tightly genetically determined component of the skull in adults; however, it is currently unknown at what point during development this pattern emerges, and whether diverse human populations follow the same ontogenetic trajectories for temporal bone growth. To address these questions, 15 ectocranial landmarks were digitized on an ontogenetic series of temporal bones from seven human populations. A MANOVA and DFA confirmed that modern human populations are significantly different in temporal bone shape even at early developmental ages, and that they can be sorted with relative reliability. Mahalanobis distances based on temporal bone shape were also significantly correlated with molecular distances among the sampled populations at each age category, suggesting that temporal bone morphology reflects genetic distances starting very early in development and continuing into adulthood. The ontogenetic trajectories were not found to differ significantly among most pairs of populations. These findings suggest that temporal bone shape diverges and reflects genetic relatedness from an early age, after which human populations develop along similar ontogenetic trajectories to reach their adult forms.

Grant Funding Source : n/a
Telomerase Biogenesis and Regulation
Authors: Baumann Peter , Tang Wen , Kannan Ram
Published: June 2021

Telomerase is the ribonucleoprotein complex that replenishes DNA sequences at the ends of chromosomes using part of an RNA subunit as template for telomeric DNA repeat synthesis. Whereas insufficient telomerase activity is associated with various degenerative disorders including dyskeratosis congenita, aplastic anemia and idiopathic pulmonary fibrosis, upregulation of telomerase is common in cancers. These observations provide substantial impetus for studying the biogenesis pathway of this RNP, how it is recruited to chromosome ends and what factors modulate its activity.

We are particularly interested in the sequence of events that lead from the transcription of the core telomerase subunits to the assembly of an active RNP. Using fission yeast as a model system we have identified a sequence of molecular events that constitute the maturation pathway for the telomerase RNA subunit. Corresponding experiments relating to the catalytic subunit are presently underway with the goal of identifying key targets for telomerase regulation.
Sensing array of radically coupled genetic biopixels
Authors: Hasty Jeff
Published: June 2021

While there has been significant progress in the development of engineering principles for synthetic biology, a substantial challenge is the construction of robust circuits in a cellular environment that is governed by noisy processes such as random bursts of transcription and translation. Such an environment leads to considerable intercellular variability in circuit behavior, which can hinder their functionality at the colony level. While quorum sensing is a promising design strategy for reducing variability through coordination across a cellular population, the length scales are limited by the diffusion time of the small molecule governing the intercellular communication. In this talk, I will discuss our recent progress in engineering the synchronization of thousands of oscillating colony “biopixels” over centimeter length scales through the use of redox signaling that is mediated by hydrogen peroxide vapor. We have used the redox communication to construct a frequency modulated biosensor by coupling the synchronized oscillators to the output of an arsenic sensitive promoter that modulates the frequency of colony‐level oscillations due to quorum sensing. For the detection of arsenic, we have developed thresholding and period modulation as sensing mechanisms. In the case of thresholding, oscillations arise when a pre‐determined low level of arsenic is present, while for period modulation the level of arsenic is proportional to the oscillatory frequency. In both cases, the coupling of thousands of colonies dramatically increases the signal to noise ratio and leads to the reliable detection of extremely low levels of arsenic. Given the repertoire of sensing capabilities of bacteria such as E. coli, the ability to coordinate their behavior over large length scales sets the stage for the construction of low cost genetic biosensors that are capable of detecting heavy metals and pathogens in the field.
Denial, acceptance, and loss of cell polarization for a stochastic model of positive feedback
Authors: Altschuler Steven , Angenent Sigurd , Jilkine Sasha , Wu Lani
Published: June 2021

Positive feedback plays a key role in the ability of signaling molecules to form highly localized clusters in cells. What prevents positive feedback from amplifying inevitable biological noise when an un‐clustered “off” state is desired? And, what limits the spread of clusters when an “on” state is desired? Here, we show that a minimal positive feedback circuit provides the general principle for both suppressing and amplifying noise: below a critical density of signaling molecules, clustering switches off; above this threshold, highly localized clusters are recurrently generated. Clustering occurs only in the stochastic regime, suggesting that finite sizes of molecular populations cannot be ignored in signal transduction networks. Regulating the density of signaling molecules provides a simple mechanism for a positive feedback circuit to robustly switch between clustered and un‐clustered states. The intrinsic ability of positive feedback both to create and suppress clustering is a general mechanism that could operate within diverse biological networks to create dynamic spatial organization.
Oxidative Nucleic Acid Modification and Demodification
Authors: He Chuan
Published: June 2021

Reversible chemical modifications on nucleic acids and proteins determine cell fates. The five bases that comprise nucleic acids „Ÿ adenine, guanine, cytosine, thymine, and uracil „Ÿ can be chemically and enzymatically modified. These chemical events can have significant biological consequences, particularly for gene expression. I will present chemical strategies we have developed to enrich and sequence novel nucleic acid modifications in mammalian genome. Several AlkB family proteins have been identified in the human genome that may mediate nucleic acids oxidation. Some of these proteins play critical roles in obesity/diabetes and various cancers. I will present our recent results that reveal the exact nucleic acid substrates and cellular function of some of these intriguing enzymes. Based on these discoveries we propose a new mode of biological regulation that depends on reversible RNA modification, for which we termed gRNA Epigenetics h.
Utilizing mTOR inhibition to study the role of mTOR signal transduction in angiogenesis
Authors: Ziegler Mary , Hughes Christopher C.W.
Published: June 2021

The growth and maintenance of normal blood vessels is disrupted in various disease processes, including malignant diseases in which tumor cells exploit the aberrant angiogenesis in order to grow and metastasize. Mammalian target of rapamycin (mTOR) is a downstream target of PI3K and is a central regulator of angiogenesis. We hypothesize that mTOR signal transduction is important in the molecular mechanism of angiogenesis. This study utilized a 3D in vitro fibrin bead model of angiogenesis and examined various mTOR inhibitors for their ability to block angiogenesis. In addition, protein and gene expression changes in human umbilical vein endothelial cells (HUVEC) grown in monolayer were examined. mTOR inhibition results in lumen loss in the 3D fibrin bead model of angiogenesis. HUVEC treated with mTOR inhibitors show loss of Akt phosphorylation at both S473 and T308 and the gene expression of VEGF and VEGFR2 are diminished. Claudin‐5 and VE‐Cadherin mRNA are reduced by rapamycin and one of the ATP‐competitive inhibitions. Claudin‐5 protein expression is inhibited by all of the mTOR inhibitors. These data suggest that mTOR plays a role in lumen formation and could be a regulator of endothelial cell junction formation. Further examination of these signaling pathways will contribute to our understanding of the molecular mechanisms underlying angiogenesis.
A role for endothelial cell‐derived Wnt5a in angiogenesis
Authors: Newman Andrew , Hughes Christopher
Published: June 2021

Wnts are secreted glycoproteins that regulate diverse biological processes. In this study, we use a complex 3D co‐culture model of angiogenesis to show that Wnts are involved in regulating in vitro angiogenesis. Specifically, we show that Wnt5a, a non‐canonical Wnt, is upregulated by endothelial cells actively undergoing angiogenesis. Moreover, using quantitative real‐time polymerase chain reaction, we find that smooth muscle cells express the Wnt5a receptors ROR2 and Fzd7 and that Wnt5a induces smooth muscle cell migration. Overall, our data indicate that endothelial cell‐derived Wnt5a may play a role in mural cell recruitment during angiogenesis.