bluuebunny/crossref_metadata_2025 huggingface dataset:
Depletion of Protein Interacting with Never in Mitosis Gene A‐1 (PIN1) decreases mammary adenocarcinoma cell growth in culture, but increases VEGF and tumor growth in mice
Authors: Schneider Ryan A , Liu Tongzheng , Huang Yongcheng , Keshvara Lakhu , Hoyt Dale G
Published: June 2021

Protein Interacting with Never in Mitosis Gene A‐1 (PIN1), a peptidyl‐proline isomerase, is overexpressed in some breast cancers and may promote tumorigenesis. Therefore, we hypothesized that depletion of PIN1 in EMT6 mammary adenocarcinoma cells would decrease their growth. A short hairpin RNA sequence targeting murine PIN1 or mutated control construct was cloned in pLKO.1, transfected into EMT6 cells, and stable cell lines were produced via puromycin selection. PIN1 levels were undetectable in knockdown cells via western blotting. Cells were seeded in 96‐well plates for 24–48H, and stained with crystal violet to measure in vitro proliferation. As expected, PIN1 knockdown decreased proliferation at 24 and 48H compared to control cells (p<0.05). The in vivo effect of PIN1 depletion was determined by transplanting these cells into the 4th mammary fat pad of BALB/c mice. Unexpectedly, PIN1 knockdown increased tumor volume nearly 2‐fold compared to control by day 28 (p<0.05). Extraction and western blotting revealed a nearly 2‐fold increase in VEGF, normalized to protein, in the knockdown tumors compared to control. These results reveal a dichotomy between growth in culture and in vivo, and suggest that tumor PIN1 status regulates host factors or responsiveness to them. Furthermore, loss of PIN1 in malignant epithelial cells may stimulate angiogenesis and growth in a PIN1‐positive host environment.
Regulation of Hypoxia Inducible Factor in Intestinal Epithelial Cells in Inflammation
Authors: Irizarry Karina , Glover Louise , Colgan Sean P.
Published: June 2021

Hypoxia and inflammation occur coincidentally in mucosal diseases. A number of studies have implicated the transcription factor hypoxia inducible factor (HIF) as a regulatory pathway under such conditions. Here, we hypothesized that inflammatory mediators may modulate HIF expression and/or function. Intestinal epithelial cells (Caco2 or T84 cells) were transfected with a luciferase reporter construct containing sequence corresponding to the erythropoietin hypoxia‐response element (HRE). Transfected cells were exposed to various concentrations of TNFα, IFNγ, IL‐4 or PGE2 in the presence and absence of hypoxia (p O2 20 torr) for 24 – 48h. HIF activity and expression were quantified by luciferase assay. Hypoxia (24h) induced HRE activity in intestinal epithelia by as much as 18±3‐fold (p<0.001). IFNγ selectively induced a concentration‐dependent repression of HIF activity under both normoxic (p<0.01) and hypoxic conditions (p<0.01). The level of inhibition by IFNγ (10ng/ml) was as much as 65±5% and 93±8% in normoxia and hypoxia, respectively, implicating a prominent and highly relevant repression by IFNγ. We conclude that HIF activity is prominently repressed by IFNγ, and since intestinal epithelial HIF has been implicated as a protective element in animal models of colitis, and the loss of HIF expression during inflammation may exacerbate inflammation.
Myeloperoxidase (MPO) delays neutrophil apoptosis and prolongs acute pulmonary inflammation
Authors: Filep Janos G. , Jozsef Levente , Pan Wanling , Kebir Driss El
Published: June 2021

We have identified MPO as a suppressor of apoptosis in human neutrophils in vitro. We investigated the impact of MPO on neutrophil apoptosis in vivo and on the resolution of neutrophil‐mediated inflammation in mice. Intravenous injection of MPO (5 nmol/kg) into conscious rats did not affect mean arterial blood pressure and the number of circulating leukocytes, whereas it significantly reduced the number of apoptotic neutrophils assessed ex vivo at 60 min post‐MPO. In a mouse model of spontaneously resolving (within 5 days) acute lung injury, intratracheal instillation of carrageenan evoked massive infiltration of neutrophils into the airspace and tissues, and alveolar edema. Intratracheal injection of MPO alone produced only minimal inflammatory cell accumulation in the airways, whereas it prolonged carrageenan‐induced inflammation. Thus, the number of neutrophils in the bronchoalveolar lavage (BAL) fluid and BAL fluid protein levels were still markedly elevated at Day 5 post‐carragenan plus MPO as compared with mice that were injected carrageenan only. The percentage of annexin‐V‐positive neutrophils in BAL fluid was significantly lower in carragenan plus MPO‐treated mice than in carrageenan‐treated mice. Our results indicate that MPO supression of neutrophil apoptosis can profoundly affect the duration of the inflammatory response and add a novel facet to the MPO biology. (Support: CIHR MOP‐64283).
Food security status, BMI, and fruit and vegetable consumption amongst Producemobile Users.
Authors: Tangney Christy C , Redfearn Kristin R , Allender Jessica C , Keim Kathryn S
Published: June 2021

Food insecurity can affect health, body weight and quality of life. Among food assistance users, the prevalence of food insecurity and its potential impact on body habitus and intake of fruits and vegetables is unclear. In this cross‐sectional study of 191 participants from 8 selected Producemobile sites in Chicago, we describe the demographics, food security status, and fruit and vegetable consumption of Producemobile patrons (18 years of age and older; 6th grade reading level). Weights and heights were measured, and all completed a 35‐item questionnaire containing the USDA 6‐item Core Food Security Module and the NCI All‐Day Fruit and Vegetable Screener.

Results:

Ethnic composition of the sample surveyed was as follows: 84% African American, 11% Hispanic and 5% Caucasian. Approximately 58% of the households were classified ‘food insecure without hunger’, 24% ‘food insecure with hunger’ and 18% ‘food secure’. More than half (53%) reported using the Producemobile monthly. Two‐thirds of the sample admitted to other forms of food assistance (i.e. food stamps, WIC coupons, school lunch program). BMI for the 46 men was 28.1 ± 6.8 kg/m2 (mean ± SD) and for 144 women, 30.6 ± 6.9. There was no difference in BMI between food secure and food insecure participants (p=0.88). Food insecure patrons reported consuming significantly fewer daily servings of fruit (1.0 ±1.9, p=0.006) and total fruit and vegetable servings (6.8 ± 8.1, p=0.03) when compared to those of food secure individuals (1.6 ± 2.2, 7.4 ± 5.1, respectively).

Conclusion:

Further research is needed to confirm whether lower intakes of fruits and vegetables are consistently observed amongst adults seeking food assistance and explore the barriers to such behaviors.
Bioreactors: Enabling Technologies for Clinical Translation
Authors: Knot Harm Johan
Published: June 2021

Bioreactors may be defined as devices used for the creation, development of engineered organs or tissues. With respect to the tissue engineering process per se bioreactors mimic/recreate important features/characteristics that the engineered organ or tissue of interest would be exposed to in the body. It can be used to mature or develop a tissue outside the human body, for subsequent implantation in vivo. The concept is to apply in vitro preconditioning in order to accelerate tissue formation/maturation/function in vivo. Bioreactors are thus recognized as an important enabling technology for tissue engineering and regenerative medicine. The rationale is that more complete tissue engineering on the “front end” will place fewer demands on the body's regenerative capacity in vivo. Bioreactors may also be utilized for preservation, storage or transportation of engineered tissues/organs. For example, bioreactors can provide the delivery mechanism for storage and transportation to the end user in order to store, transport and deliver the clinical product to the physician/surgeon for implantation.
Systematic analysis of cell signaling during planarian tissue regeneration, remodeling & homeostasis
Authors: Gurley Kyle Alan , Rink Jochen C , Alvarado Alejandro Sánchez
Published: June 2021

We have initiated a systematic effort to define the roles of highly conserved signaling pathways in the planarian Schmidtea mediterranea, an organism with astounding regenerative capabilities. Following amputation, freshwater planarians properly regenerate a head or tail from the resulting anterior or posterior wound. The mechanisms that differentiate anterior from posterior and direct the replacement of the appropriate missing body parts are unknown. We report that RNA interference (RNAi) of β‐catenin or dishevelled causes the regeneration of a head instead of a tail at posterior amputations. Conversely, RNAi of the β‐catenin antagonist adenomatous polyposis coli (APC) results in the regeneration of a tail at anterior wounds. The reversal of tissue identity is complete by both anatomical and molecular criteria, it can be observed as early as 12 hours post‐amputation, and it is independent of the angle or location of amputation. However, systematic silencing of known upstream components did not recapitulate these defects. The intracellular components of canonical Wnt signaling may therefore be regulated by an unconventional upstream mechanism in regenerating planarians. In addition, β‐catenin RNAi transforms the tail of uncut adult animals into a head. We suggest that β‐catenin is a molecular switch that specifies and maintains AP identity during planarian regeneration and homeostasis.

K.A.G., F32GM082016, T32CA093247. J.C.R., European Molecular Biology Organization. A.S.A., HHMI Investigator and RO‐1 GM57260
Migrating the adult brain: Vascular and ECM mediated?
Authors: Puche Adam C
Published: June 2021

New cells are continuously added to the rodent olfactory bulb (OB) throughout development and in adult. These cells migrate tangentially from the sub‐ventricular zone along the rostral migratory stream to the OB where they migrate radial from the centre to periphery of the OB. In adult, this tangential migration takes place in ‘chains’ of cells contained within glial tubes, in contrast, radial migratory cells lack these defined glial tubes and chains, migrating instead as individual cells. Although different modalities of radial migration have been described in other brain regions, the mechanisms governing radial migration in the OB are still largely unknown. Here we identify a modality of migration in which neuronal precursors migrate along blood vessels toward their destination. Our results show that many radially migrating cells associate with the vasculature in the granule cell layer of the OB and in vivo time lapse imaging demonstrates that they use blood vessels as scaffold for their migration through an interaction with the extracellular matrix and perivascular astrocytes end feet. Indeed, matrix remodeling appears important for normal migration in this pathway.
Molecular and biophysical regulators of lymphatic transport
Authors: Swartz Melody Ann , Rutkowski Joseph , Miteva Dimana O , Issa Amine , Dixon James Brandon
Published: June 2021

Although the lymphatic system plays critical roles in tissue fluid balance, lipid transport, and immune cell trafficking, the active regulation of its drainage function, particularly in lymphatic capillaries, is poorly understood. Here we present in vitro and in vivo studies that reveal how lymphatic endothelial cells (LECs) actively regulate their function. In vivo, we explore how lipid transporters and diet affect lymphatic function in a series of transgenic mouse models. In vitro, we explore functional‐adaptive responses of lymphatic endothelium to luminal and transmural flow and to lipid mediators. Specifically, LECs respond to shear stress by upregulating fluid and solute transporters, increasing their permeability, altering many lipid metabolites, and downregulating cell proliferation, among others. This indicates that during lymphedema or fluid stagnation, the potential for fluid and solute transport is drastically compromised, lipid metabolism is altered, and vessels become hyperplastic, which corresponds to in vivo observations. Lipid mediators also have striking effects on lymphatic function. This work reveals key molecular regulators of lymphatic function and shows how shear stress, an indicator of functional state, as well as lipid mediators, affects those regulators. This work also supports the notion that lymphedema and obesity are intimately correlated.
The regulation of stem cell self‐renewal
Authors: Morrison Sean J.
Published: June 2021

The mechanisms that regulate stem cell self‐renewal are fundamental to the formation and maintenance of normal hematopoiesis as well as hematopoietic malignancies. Fetal stem cells differ phenotypically and functionally from adult stem cells in diverse tissues. However, little is known about how these differences are regulated. To address this we compared the gene expression profiles of fetal versus adult hematopoietic stem cells (HSCs) and discovered that the Sox17 transcription factor is specifically and preferentially expressed in fetal and neonatal but not adult HSCs (Cell 130:470). Germline deletion of Sox17 led to severe fetal hematopoietic defects, including a lack of detectable definitive HSCs. Conditional deletion of Sox17 led to the loss of fetal and neonatal but not adult HSCs. Sox17 expression by HSCs was extinguished three to four weeks after birth as HSCs acquired adult properties. During this transition, loss of Sox17 expression by individual cells correlated with slower proliferation and the acquisition of an adult phenotype. Sox17 thus distinguishes the transcriptional regulation of fetal and neonatal HSCs from that of adult HSCs.

As HSCs enter the postnatal period, they become dependent upon the polycomb family transcriptional repressor, Bmi‐1. Bmi‐1 is required for the postnatal maintenance of every type of stem cell yet examined including HSCs (Nature 423:302, 2003) and neural stem cells (Nature 425:962, 2003). Bmi‐1‐deficient mice die by early adulthood with stem cell depletion, growth retardation, ataxia, and seizures. These phenotypes correlate with increased expression of the cyclin‐dependent kinase inhibitor p16Ink4a and the p53 agonist p19Arf, which promote cellular senescence. Deletion of Ink4a and/or Arf from Bmi‐1−/− mice partially rescues stem cell self‐renewal and stem cell frequency (G&D 19:1432, 2005). Despite ongoing Bmi‐1 expression, Ink4a expression increases with age, reducing stem cell frequency and function (Nature 443:448). The regulation of stem cell self‐renewal throughout life by networks of proto‐oncogenes and tumor suppressors emphasizes the mechanistic links between stem cell self‐renewal and cancer cell proliferation.
Genome‐Wide Maps of the Human Transcriptome Reveal an Interleaved Organization and Novel Short and Long Classes of RNAs
Authors: Gingeras Thomas R
Published: June 2021

Genomic regions not coding for proteins nor involved in cis or trans‐acting regulatory activities are at best viewed as sites for evolution‐mediated experimentation of novel functional domains and at worst, “junk”. However, recent unbiased empirical genomic wide have revealed that there are large portions of the non‐protein coding are transcribed in a regulated fashion in several organisms including humans. A recurrent question derived from these recent findings centers upon the biological functions of the observed unannotated transcripts. Considerable portions of the unannotated transcription observed in these species serve as unreported parts of protein coding transcripts and as precursors to the generation of short RNAs. As part of the Encyclopedia of DNA Elements (ENCODE) project, studies reveal that more than half of the well characterized protein coding loci present in ENCODE designated human genome regions (∼400) make use of tissue specific alternative, unanntoated 5′ transcriptional start sites (TSS) which for as much as 30% of the extended transcripts, are more than 200,000 nucleotides (nt) upstream of the annotated portions of the transcripts (108,000 nt on average). We have made similar observations in the organization and regulation of transcription during the first 24 hours of development of Drosophila melanogaster. In addition, often sharing the same genome sequences as protein coding transcripts are distinct l nuclear transcripts which serve as primary transcripts for the formation of small RNAs ranging in size from∼20–200 nt. Recently, two new classes of short RNAs have been identified from our analyses of the sites of transcription across the human genome. These data support a new model for genome organization which is not co‐linear but rather lattice‐like and in part focused on the production of short RNAs.
Surveying regulatory feedback among yeast transcription factors
Authors: Brem Rachel B
Published: June 2021

Complex quantitative cellular behaviors—such as binary switching, signal amplification, and adaptation—are at the heart of systems biology. To date, most such behaviors have been characterized by tour de force experiments on a handful of model pathways. The full spectrum of pathways in which nature has quantitatively tuned the signaling machinery is almost unknown. Many signaling behaviors are thought to require regulatory feedback, in which a protein influences its own expression or abundance. With the goal of using feedback as a signpost for signaling behaviors of interest, we have surveyed the prevalence of this motif across transcription factors in S. cerevisiae. We find evidence that direct feedback, in which a transcription factor protein binds upstream of its own gene, and indirect feedback, mediated by multiple signaling intermediates, are both common. We also find that positive and negative feedback, respectively the up‐ or down‐regulation of abundance of a transcription factor by its own action, are both prevalent. And we observe cases of an unusual feedback model in which a transcription factor regulates its own subcellular localization. Our results shed light on the functional genomics of feedback, and they identify particular yeast circuits as novel, testable candidates for binary switches and other systems properties.
Promoter architecture dynamically modulates transcription factor binding and transcriptional output over time
Authors: Lieb Jason Dillon , Hanlon Sean , Lickwar Colin
Published: June 2021

Our laboratory studies relationships between chromatin, transcription factor binding, and transcriptional output in S. cerevisiae, C. elegans and human cells. Here, I will present a study in which we experimentally determined the genomic distribution of the yeast transcription factor Rap1 during vegetative growth, respiratory growth and throughout meiosis and sporulation in yeast. Simultaneously, we monitored the expression of the genes downstream of binding events. In short, all simple combinations of binding dynamics and transcriptional output were observed. Even though within the same cell individual targets downstream of Rap1 binding may be strongly repressed or strongly activated, we demonstrate that Rap1 is required for both types of transcriptional outcome by depleting the transcription factor from cells. While there is no simple relationship between Rap1 binding and transcriptional activity, differences in transcriptional outcome can be partially explained by the presence of additional transcription factor binding motifs and the type of Rap1 motif contained within the promoter of target genes. We are also developing a system in yeast to probe transcription factor binding dynamics genome‐wide in yeast, and will report progress in that area as well.
oskar RNP assembly for coordinated transport and translation control
Authors: Ephrussi Anne
Published: June 2021

oskar encodes the Drosophila posterior determinant, whose accumulation at the posterior of the oocyte is necessary for abdomen and germline formation in the embryo. Spatial restriction of Oskar is critical and is achieved by coordination of mRNA localization and translational control: unlocalized oskar mRNA is repressed and the mRNA translated only upon posterior localization. oskar mRNA localization requires Exon Junction Complex (EJC) proteins and splicing of the first intron. The 3′UTR is also required for posterior targeting of the mRNA, and promotes co‐assembly of oskar 3′UTR‐containing transcripts into transport complexes. Translational repression of oskar is mediated by Bruno protein, which binds to the 3′UTR and represses translation via two mechanisms: one, cap‐dependent, in which binding of eIF4G and small ribosomal subunit recruitment are inhibited; the other, cap‐independent, in which oskar mRNA is oligomerized into large particles, also blocked at initiation. New insight into the importance of oligomerization in oskar mRNA regulation, as well as the significance of EJC positioning upstream will be discussed.
TPI deficiency and neurodegeneration in Drosophila: an AGE‐dependent link to a common mechanism?
Authors: Ganetzky Barry , Gnerer Joshua P. , Kreber Robert A.
Published: June 2021

We have conducted forward screens in Drosophila to identify genes required for maintenance of neuronal viability. wasted away (wstd), is defined by a hypomorphic, autosomal recessive mutation associated with progressive motor impairment, severely reduced lifespan, and vacuolar neuropathology. Molecular analysis demonstrates that wstd encodes the glycolytic enzyme, triosphosphate isomerase (TPI). Mutations causing TPI deficiency have also been identified in humans resulting in progressive neurological dysfunction, neurodegeneration and early death. However, the mechanistic basis of the human syndrome remains puzzling. Decreased ATP levels do not appear to cause the observed phenotypes in TPI deficient flies or humans because ATP levels remain normal. Moreover, we find no genetic evidence that the mutant TPI is misfolded or involved in aberrant protein‐protein associations. We propose instead that the enzymatic block in wstd mutants leads to an accumulation of methylglyoxal and the consequent enhanced production of advanced glycation endproducts (AGEs), which are ultimately responsible for death and dysfunction of TPI deficient neurons. Evidence in support of this interpretation will be described. Our results favor the idea that TPI serves an essential neuroprotective role and support the possibility that AGEs may also contribute to the pathogenesis of other human neurological disorders.
Axonal protection by Nmnat expression
Authors: Milbrandt Jeffrey
Published: June 2021

Axonopathy is a critical feature of many nervous system disorders including peripheral neuropathies and neurodegenerative diseases like ALS and Parkinson's disease. Studies of Wlds mutant mice have demonstrated that axonal degeneration is an active process. This degenerative process can be altered by manipulating neuronal bioenergetic pathways and increased expression of enzymes involved in NAD biosynthesis. Targeting of the Nmnat molecule to the cytoplasm, mitochondria, or the nucleus all result in substantial axonal protection. Axonal protection by Nmnat expression protects axons against a variety of insults including mechanical injury, microtubule disruption and mitochondrial inhibitors, and is dependent on its NAD biosynthetic activity.
Rho GTPase signaling during polarized cell migration
Authors: Hall Alan
Published: June 2021

The actin and microtubule cytoskeletons play a central role in defining cell shape and polarity and in driving many of the dynamic aspects of cell behavior such as cell migration, axon guidance, phagocytosis and cell division. Since, a major function of Rho family GTPases is to control the assembly, disassembly and organization of actin and tubulin filaments, they play an important part in regulating these cell biological processes. Cdc42 was first identified in budding yeast as an essential regulator of polarity during cell division and in pheromone‐dependent mating. It has since been shown to play a key role in establishing cell asymmetry in higher eukaryotes. Using primary rat embryo astrocytes and fibroblasts in in vitro assays, we have found that Cdc42 controls the polarity signals that are required for directed migration. Cdc42 controls the polarity of the microtubule cytoskeleton through a spatially localized signal transduction pathway involving: (i) the Par6/aPKC complex, (ii) inactivation of GSK‐3 and (iii) the association of the tumour suppressor APC with microtubule plus ends. We are currently characterizing the details of this signal transduction pathway to understand the biochemical mechanism that control cell polarity during migration.
Spatiotemporal dynamics of lipid second messenger signaling
Authors: Newton Alexandra C
Published: June 2021

Protein kinases B (Akt), C, and D are regulated by lipid second messengers: 3’phosphoinositides for protein kinase B and diacylglycerol for protein kinases C and D. Using genetically‐encoded reporters for each of these kinases, we have determined the spatiotemporal dynamics of lipid second messenger signaling mediated by each of these kinases at defined intracellular locations. Although all three kinases are rapidly activated at the plasma membrane, we find that all three kinases have robust and much more sustained activity at the Golgi. In addition, tethering of the reporters to specific scaffolds for each of these kinases reveals highly localized activation within the microenvironment of the scaffold. These results underscore the importance of subcellular location and scaffolding in lipid second messenger signaling.
Assessment of parent and child perceptions of rules and expectations regarding beverage intake
Authors: Roth‐Yousey Lori , Asche Kim , Schroeder Mary , Reicks Marla
Published: June 2021

Beverage consumption patterns of children influenced weight outcomes in previous studies. Two evaluation instruments were tested in parent‐child dyads (n=79) to assess perceptions of rules and expectations regarding beverage intake. Child (n=28) and parent statements (n=26) were constructed and children (mean age 11.5 years, 53% boys, 81% white) and parents (91% 31–50 year age range, 81% women, 91% white) separately rated their agreement with statements on a 5‐point scale. Principal components analysis resulted in 5 child scales and 6 parent scales. Child scales included meal‐ (4 items, α=.80) and beverage‐specific rules (sweetened beverages, 4 items, α=.88; milk, 2 items, α=.78), parent communication (3 items, α=.74), offering choices by permission (3 items, α=.74) or availability (3 items, α=.53). Parent scales included permissiveness (2 items, α=.70), limiting sweetened beverage intake (5 items, α=.86), encouraging milk intake (5 items, α=.78), role modeling (3 items, α=.73), and family member influences (3 items, α=.66). Correlations with parent and child intake of milk, soft drinks, fruit drinks and orange juice, as measured by a food frequency questionnaire, confirmed that the scales were measuring parent or child constructs related to beverage intake. These instruments can be used to evaluate post‐intervention changes in rules/expectations for beverage intake in parents and children.
Effects of increased dietary protein and meal patterning on appetite during short‐term energy balance and energy restriction
Authors: Leidy Heather J , Bossingham Mandi J , Carnell Nadine S , Mattes Richard D , Campbell Wayne W
Published: June 2021

Objective: This study assessed whether the timing of increased dietary protein intake throughout the day influences the feeling of fullness during energy balance (EB) and restriction (ER). Methods: 9 men (age: 45 ± 5 y; BMI: 32.5 ± 0.6 kg/m2) randomly completed 5 controlled feeding trials consisting of 3 days each of EB and ER (750 kcal/d reduction). The diet was composed of a normal amount of protein (NP) (0.8 g protein·kg−1·d−1) or an additional amount of protein (HP) (+0.6 g protein·kg−1·d−1) given at breakfast (B), lunch (L), dinner (D), or equally divided among all meals (E). Feelings of fullness were assessed over 15 h. Results: No difference in fullness was observed between the EB vs. ER trials; thus, the data were combined. With all HP meals combined, HP led to higher fullness (P<0.05) across the day vs. NP. When comparing the timing of increased protein intake, the 3‐h postprandial, subsequent meal, evening, and overall fullness were not different between HP‐L, HP‐D, or HP‐E vs. NP but were higher for HP‐B vs. NP (p<0.05 for all effects except evening (p=0.1)). Comparing the HP treatments, the HP‐B led to greater overall fullness vs. HP‐L (p<0.005) and HP‐D (p<0.01) but not for HP‐E. Conclusion: Increased dietary protein provided at breakfast leads to immediate and sustained increases in feelings of fullness in overweight men.

Support: National Pork Board; American Egg Board; IBRC, Purdue University
Evidence for a long time course adaptation of glucose metabolism to high protein feeding in rats without changes in resting energy expenditure
Authors: Stepien Magdalena , Azzout‐Marniche Dalila , Even Patrick , Payet Audrey , Simonin Angélique , Fromentin Gilles , Tomé Daniel , Gaudichon Claire
Published: June 2021

High protein (HP) diet decreases spontaneous energy intake in rat. This effect has been suspected to be associated to an increased thermic effect of feeding (TEF). We evaluate the changes of energy expenditure and nutrient oxidation occurring during the adaptation to a HP diet. For that purpose 48 Wistar rats were fed a normal protein (NP) diet for 1 week and then a HP diet. Energy metabolism was measured in an open circuit indirect calorimetry after 1, 3, 6 or 15 d. After fasting overnight, they received a calibrated meal of 4 g. The postprandial metabolism was followed for 4 h. During the fasting state, total but not resting energy expenditure (REE) tended to increase in HP groups (p=0.06). Activity differed among groups with a trend to increase from 1 to 6 d of HP diet. Compared to NP, TEF increased as soon as the second day of HP diet and was the highest after 6 d. During fasting, there was a trend to a lower fat oxidation after 3 d of HP diet, at the expense of protein oxidation. In the fed state, fat oxidation increased after 1 d of HP diet but then decreased after 3 d. Interestingly, CHO oxidation was twice lower after 1 d of HP diet and then progressively increased until 15 d but remained lower than in NP rats. We showed that TEF and activity but not REE were enhanced by the HP diet. This also led to dramatic changes in postprandial nutrient oxidation and suggested a slow adaptation of CHO metabolism from 3 to 15 d.
Offering free fresh fruits and vegetables during the school day affects students’ fruit and vegetable intake but is dependent upon implementation strategies
Authors: Shertzer Julie Anne , Fly Alyce D. , Middlestadt Susan E. , Hightower‐King Mindy
Published: June 2021

Most US youth eat too few fruits and vegetables (FV). Thus, in 2004, a federally funded program was established in 8 states to improve intake by offering free FV at schools, mainly in classrooms. Evaluation of Indiana program efforts to increase FV availability and improve intake of students in 4th–8th grades for the 06–07 school year was conducted by choosing 6/25 schools by convenience, administering questionnaires to teachers and students, and examining FV purchase records. Teachers (n=142) reported participation by internet survey; differences were detected using Kruskal‐Wallis with Mann‐Whitney post‐hoc tests. Students reported “yesterday” intake (portions/day) and demographics at baseline (n=1,227) and follow‐up (n=1,051). Data were analyzed using t‐tests and Chi Square, respectively. Significance was set at α=.05. Offerings varied among schools but overall were 81.6% fresh F, 15.8% V, 2.6% dry F. Student participation (94%) was high. Teacher participation (96%) was high but varied by school (p=.032). At follow‐up, students improved whole F intake (1.7±1.1 vs 1.2±1.1; mean portions ± SD; p<.001) but significance was limited to 4/6 schools; those at schools serving >20% V increased intake of certain V (p=.004). Availability of fresh FV improves intake but depends on implementation. Schools should offer more V to improve V intakes. Supported by Indiana Department of Education & Indiana University.
Oxidative Stress in Premature Infants
Authors: Friel James Kenneth , Chiu Aaron , Diehl‐Jones William , Roberts Jackson
Published: June 2021

We hypothesized that early infant mortality in the premature infant (< 37 weeks gestation), which is normally high, as well as increased morbidity reflects the difficulty of adaptation to oxygen. Therefore forty PT infants (birth weight: 1146 ± 38 g; gestational age 28 ± 0.3 wks) were followed biweekly once oral feeds were introduced. Oxidative stress was quantified by measuring dinor, dihydro metabolites of F2‐isoprostanes and isofurans in urine by liquid chromatography/tandem mass spectrometry. The precision of the assay is ± 5% and the accuracy is 80%. For F2‐isoprostanes results were : 0–2 wks, 147 ± 12; 3–4 wks, 213 ± 34; 5–6 wks, 203 ± 16; 7–8 wks, 246 ± 38; 9–10 wks, 240 ± 38; > 10 wks, 191 ± 47 (ng/mg creatinine, mean ± SE). The normal levels from 40 adults are 38.1 ± 19.1 ng/mg creatinine. For isofurans: 0–2 wks, 17 ± 2; 3–4 wks, 20 ± 3; 5–6 wks, 24 ± 3; 7–8 wks, 24 ± 5; 9–10 wks, 25 ± 5; > 10 wks, 31 ± 8 ng/mg creatinine. The normal levels in adults (n=22) are 4.35 ± 1.9 ng/mg creatinine. Ability to resist oxidative stress was assessed with ORAC: baseline: 6.6 ± 0.2 and at discharge, 6.7 ± 0.1 (healthy premature 6 mos, 6.6 ± 1.1; full term 4 & 12 M, 9.9 ± 0.6; adult 9.3 ± 0.9. These data suggest that the PT human infant is under severe oxidative stress and it may be appropriate to consider routine antioxidant supplementation for the first months of life.
Comparison of energy intake and satiety for whole grain barley, wheat and refined rice
Authors: Schroeder Natalia , Gallaher Dan , Arndt Beth , Marquart Len
Published: June 2021

This study compared the effect of whole grain high‐fiber barley, whole grain wheat and refined rice on energy intake and satiety. Fifty healthy subjects were recruited for this crossover design. Subjects consumed a hot cereal and a snack mix followed by an ad lib lunch once per week. Energy intake was measured through pre and post plate waste as part of the smorgasbord lunch. Through the use of a visual analog scale, hunger and fullness were measured for within and between treatment differences. Data analysis was completed using Proc mixed for between treatment differences and proc t‐test for within treatment differences. Energy intake at lunch did not significantly differ between treatments (p<0.8626). Satiety differed between and within treatments. Subject's felt significantly less hungry on the barley compared to the wheat (p<0.0208). Additionally, the subjects reported significantly less hunger before lunch compared to baseline (p<0.0017) for barley, but not for wheat (p<0.2085) or rice (p<0.0717). The greatest fullness was experienced on the barley compared to the wheat (p<0.0530). Compared to baseline, subjects felt the fullest before lunch on the barley followed by the wheat, and rice. Longer term studies are needed to assess the impact of whole grain foods on energy intake, satiety and weight management.

ConAgra Foods Inc.
Heart disease knowledge and dietary intakes of African Americans aged 18–30 years
Authors: Winham Donna M , Jones Kathleen M.
Published: June 2021

Heart disease knowledge and dietary intakes were assessed with a convenience sample of 192 African Americans aged 18–30 years in Arizona. The Jackson Heart Study food frequency questionnaire and a validated heart disease knowledge instrument were self‐administered. Participants had a mean age of 22.1± 3.7 years, an average BMI of 27.4 ± 6.2, and 70% had at least 1 year of college education. Forty percent of the sample was male. College educated participants consumed slightly more servings of fruits and vegetables daily (2.3± 1.5 vs. 2.0 ± 1.6; p=.248), and less mean fat intakes (g/d) (21.1± 7.5 vs. 25.1 ± 8.8; p=0.003) than non‐college educated participants. Recognition of some prevention measures such as reducing dietary cholesterol (79%), and lowering salt intakes (85%) were high, but other concepts such as reducing dietary animal products (45%) and taking multivitamins (46%) were less known across education, gender and age categories. Women scored higher than men on heart disease knowledge (p=0.001), but awareness that women had unique heart disease risks was low for both genders (53%). Heart disease prevention programs should focus on dietary improvements in this age group of African Americans. Specific examples are knowledge of dietary cholesterol in the diet (e.g. animal products), and the atypical heart disease symptoms found in women, among others. Funding provided by St. Luke's Health Initiatives.
Dynamics of adolescent and adult beverage intake patterns in Mexico
Authors: Barquera Simon , Hernandez‐Barrera Lucia , Tolentino‐Mayo Maria Lizbeth , Espinosa Juan , Ng Shu Wen , Rivera Juan Angel , Popkin Barry
Published: June 2021

Objective:

To examine the patterns and trends in adolescent and adult beverage intake in Mexico.

Design:

Use of nationally representative dietary intake, income, and food expenditure surveys.

Subjects: For the nationally representative surveys adolescents (aged 12–18y) and adults (aged 19y and older) were studied. For the 1999 and 2006 national nutrition surveys, there were 416 and 7,464 adolescents and 2180 and 21,113 adults, respectively, available in the overall sample.

Measurements: Volume (ml) and energy (kcal) per day contributed by all beverages consumed by the subjects was measured.

Results:

The trends from both the food expenditure and the dietary intake surveys showed large increases in caloric beverages among adolescents and adults over the past decades—particularly 1999–2006. Coupled with very high‐income elasticities, this showed the high likelihood that intakes will increase as Mexican incomes continue to rise. Further, the price elasticities for whole milk and sodas were both greater than 1 and triple those for all other food groups.

Conclusion:

Mexican adolescents and adults consumed 20.1 and 22.3% of their calories from beverages. Whole milk, sugared‐beverages, fruit juice and alcohol represented the four major categories of beverage intake.
Dietary intake of iron and zinc by urban Guatemalan schoolchildren in the context of food pattern modernization
Authors: Montenegro‐Bethancourt Gabriela , Vossenaar Marieke , Doak Colleen M. , Solomons Noel W.
Published: June 2021

Background:

Meeting iron (Fe) and zinc (Zn) needs is related to foods consumed and nutrient bioavailability.

Objective:

To estimate the dietary intake of Fe and Zn and their adequacy among 3rd and 4th grade schoolchildren in Quetzaltenango, Guatemala.

Methods:

Data were obtained in a cross‐sectional study of 449 students, boys and girls, stratified by SES in private (n=230) or public (n=219) schools. Intake estimation comprised a single 24‐h pictorial self‐recording, supplemented by a dietitian interview to quantify amounts. Adequacy reference was the 2004 WHO recommended nutrient intakes (RNI).

Results:

Median intake of Fe, 12.9 mg, represented 145% of its RNI. Its 5 main sources were: fortified cornflakes; sweet rolls; fortified Incaparina gruel; beefsteak; and fried eggs. Median intake of Zn, 7.9 mg, represented 141% of RNI. Its 5 main sources were: beefsteak; fortified Incaparina gruel; corn tortilla; ground beef; and fried eggs. The leading repast for Fe was breakfast (36.5% of total), whereas lunch led for Zn (35.2%). Intake of Fe (p<0.001), but not of Zn (p=0.851), was higher in the private school sample.

Conclusions:

Intakes of Fe and Zn are generally adequate in this western highlands city, and supported, in part, by fortified foods.
Enhanced liver regeneration in transgenic mice overexpressing stable β‐catenin
Authors: Nejak‐Bowen Kari , Thompson Michael , Monga Satdarshan P
Published: June 2021

Previously, our lab demonstrated an essential role of β‐catenin in liver regeneration. In order to elucidate the effect of β‐catenin overexpression on regeneration, we utilized recently generated transgenic (TG) mice that express a constitutively active form of β‐catenin in regeneration studies and found that constitutively active β‐catenin leads to an increase in cell proliferation after partial hepatectomy (PHx). In TG mice, proliferation (as measured by PCNA immunohistochemistry) was increased over wild‐type (WT) mice at all time points analyzed. At 40H after PHx, before the onset of proliferation in WT FVB mice, almost every hepatocyte in the TG liver stains positive for PCNA. This proliferative increase in TG mice extends to D14; while PCNA in the WT is near resting levels, TG mice continue to show increased PCNA staining. Analysis of cyclin D1 expression by Western blotting showed that levels of this cell‐cycle regulator are approximately equal in both WT and TG mice 10 hours after PHx. However, at 40H, cyclin D1 levels in TG mice are several‐fold higher than WT. Conversely, protein levels of glutamine synthetase (GS), a known downstream target of β‐catenin which is not involved in the cell cycle, remains unchanged between TG and WT. In conclusion, β‐catenin overexpression causes an upregulation of downstream targets such as cyclin D1, which causes an increase in proliferation during regeneration.
Analysis of gene expression patterns of hepatocyte‐like cells derived from human bone marrow mesenchymal stem cells
Authors: Stock Peggy , Staege Martin S , Müller Lutz P , Volkmer Ines , Lützkendorf Jana , Christ Bruno
Published: June 2021

The aim of the study was to determine the degree of differentiation of hepatocyte‐like cells derived from human mesenchymal stem cells (hMSC) by comparing global gene expression patterns of differentiated and undiffentiated hMSC from bone marrow (hBM‐MSC), adult liver and fetal hepatocytes.

hMSC were isolated from bone marrow and differentiated into hepatocyte‐like cells. Microarray analysis was performed using the HG‐U133A array from Affymetrix. For comparative analysis of expression patterns published datasets and results obtained from this study were combined. All datasets were analysed by Microarray Suite 5.0 and Genesis software.

In comparison to undifferentiated hBM‐MSC, hepatocyte differentiated hBM‐MSC showed a dramatic change of the gene expression profile. We found approximately 1000 genes upregulated and 1400 genes were downregulated in differentiated MSC with a fold change ≥3. Among these, around 60% and 40% showed similar expression levels as in adult liver, respectively. Expression of glutamine synthetase and ribonuclease 4 is significantly upregulated in hepatocyte differentiated MSC, typical for adult hepatocytes in the perivenous area of the liver lobule. Thus, hepatocyte‐like cells derived from hBM‐MSC feature human hepatocyte gene expression patterns, which encourages to step forward to use hBM‐MSC as a novel source for cell therapy of liver diseases.
Effect of citrulline on cultured macrophages from Zucker Diabetic Fatty (ZDF) rats
Authors: Bonhomme Sandra , Kohler Josephine , Dumez Celine , Breuillard Charlotte , Leplenier Servane , Couderc Remy , Cynober Luc , Darquy Sylviane
Published: June 2021

Obese and type 2 diabetic patients present metabolic and immune impairments that weaken response to aggression and infectious complications. Arginine (Arg), which level is depressed in diabetes, has immune properties and is essential in situations of stress. In particular, it is the precursor of nitric oxide (NO). Since administration of Arg is associated with adverse effects, the use of citrulline (Cit) as an indirect progenitor for NO is of particular interest.

Objectives:

To evaluate the effect of Cit on macrophages from ZDF control fa/+ and diabetic‐obese fa/fa rats.

Peritoneal macrophages (5105 cells) from fa/+ and fa/fa rats were cultured with 0.5, 1 and 2 mmol/l Cit concentrations for 24 h (n=12/group). NO and TNFα were measured in the supernatant. In cell extracts, iNOS protein expression was evaluated by Western blot.

Results:

In the fa/+ rats NO production was related to Cit concentration, and TNFα concentration remained constant. In fa/fa rats NO production increased with 1 mmol/l of Cit, and TNFα production decreased with high Cit concentration.

Conclusions:

Cit can act as a precursor for Arg in macrophage NO production. Like arginine, Cit has an anti‐inflammatory effect by decreasing TNFα production and is able to restore NO production in inflammatory situations such as diabetes and obesity. Cit administration can be considered as an alternative to arginine supplementation in clinical use.
The transcription factor Erg regulates angiogenesis and endothelial apoptosis
Authors: RANDI ANNA MARIA , Birdsey Graeme , Dryden Nicola , Amsellem Valerie , Gebhardt Frank , Haskard Dorian , Dejana Elisabetta , Mason Justin Charles
Published: June 2021

Tight regulation of the balance between apoptosis and survival is essential in angiogenesis. The Ets transcription factor Erg is required for angiogenesis. Inhibition of Erg expression in human umbilical vein endothelial cells (HUVEC) using antisense resulted in detachment of cell‐cell contacts. A key regulator of junction stability in EC is VE‐cadherin. Using chromatin immunoprecipitation (ChIP), we found that Erg binds the VE‐cadherin promoter in HUVEC; moreover, inhibition of Erg expression in HUVEC resulted in a significant decrease in VE‐cadherin levels. Both Erg and VE‐cadherin provide survival signals to EC; apoptosis induced by inhibition of Erg was partly rescued by over‐expression of VE‐cadherin‐GFP, suggesting that VE‐cadherin is involved in the Erg‐dependent survival signals. Additional survival signals mediated by Erg may be via the anti‐apoptotic molecule Bcl‐2, since Erg binds the Bcl‐2 promoter and inhibition of Erg expression in HUVEC results in decreased Bcl‐2 levels. To validate the role of Erg in angiogenesis in vivo, we used Erg siRNA in a Matrigel plug model. Matrigel, mixed with bFGF and either Erg or control siRNA, was injected subcutaneously in mice. Erg inhibition resulted in a significant decrease in vascularization of the plugs, as measured by H&E and CD34 staining. These results identify a new pathway regulating angiogenesis via Erg, VE‐cadherin and endothelial survival.

Research supported by the British Heart Foundation
Cardiac effects of pulmonary influenza A viral infection
Authors: Bradbury J Alyce , Carey Michelle A , Seubert John M , Myers Page , Rouse Clay , Edin Matt , DeGraff Laura M , Germolec Dori , Zeldin Darryl C
Published: June 2021

Rationale: Clinical studies have shown a causal relationship between influenza A viral infection (IAV) and cardiovascular disease. Methods: C57BL6 mice were infected intranasally with 200 pfu of Hong Kong influenza A (H3N2) virus while control mice received intranasal saline. Body weight and temperature were recorded daily as indicators of morbidity. On day 4, echocardiograms were performed to assess cardiac dimensions and fractional shortening. Hearts were isolated and perfused in a Langendorff mode to measure contractile function (LVDP). Cytokines and protein kinases were analyzed using Bioplex arrays. Results: IAV induced significant weight loss and changes in body temperature. Echocardiograms showed a reduction in % fractional shortening in the IAV group compared to control (p<0.05). In perfused hearts, there was an increase in post‐ischemic LVDP in IAV vs. controls (p<0.05). Serum IL‐6 levels were elevated on days 2–4, serum TNF‐αlevels were elevated on day 2, phospho‐Akt (pSer473) was suppressed on day 1, and phosphorylation of p38 MAPK (pThr180/pTyr182) was increased on day 4 in IAV group relative to controls (all p<0.05). Conclusion: We postulate that improved functional recovery may be due to a preconditioning‐like effect on the heart induced by IL‐6 and TNF‐α. This research was supported by the Intramural Research Program of the NIH, NIEHS.