bluuebunny/crossref_metadata_2025 huggingface dataset:
The associations between serum antioxidant concentrations and bone mineral density in women aged 50 and over: an analysis of NHANES 2005–6
Authors: Hamidi Maryam Sarah , Cheung Angela Man‐Wei
Published: June 2021

It has been shown that diets high in antioxidants are associated with high bone mineral density in postmenopausal women. Conversely, some studies suggest that dietary supplements containing antioxidants may have detrimental effects on bone. Serum concentrations of antioxidants can reflect their intakes from both food and supplements. Thus, the objective of our study was to examine the associations between serum concentrations of several antioxidants and femoral neck BMD (FN_BMD) in postmenopausal women aged ≥50 years. We used cross‐sectional data from the NHANES 2005–6. Multiple regression models with adjustments for relevant confounders were used to examine the associations between serum concentrations of vitamins C and E (alpha‐ and gamma‐tocopherol) and alpha and beta‐carotene, and FN_BMD. The study sample included 168 women, with a mean age of 67.5±0.5 years, who were free from diseases and medications that affect bone metabolism, and were fasting >;9 hours prior to examination. In adjusted models, serum concentration of vitamin C had a positive association with FN_BMD (p=0.018), while serum concentration of beta‐carotene had a negative association with FN_BMD (p=0.008). The association between higher serum vitamin C and higher BMD in our study is consistent with findings of other studies. The negative association between serum beta‐carotene and femoral neck BMD needs further investigation.

Grant Funding Source: None
Weight change, calorie intake, and macronutrient profiles for 6 and 12 month diet and exercise interventions in participants with pre‐diabetes: Does duration of treatment make a difference?
Authors: Granville Esther O , Starr Kathryn N , Ratliff April , Connor Lynn A , Slentz Cris A , Bateman Lori A , Willis Leslie H , Piner Lucy , Kraus William E , Bales Connie W
Published: June 2021

Dramatically increasing rates of type 2 diabetes (T2D) underscore the need for successful long‐term approaches that halt progression of pre‐diabetes to T2D. We examined body mass (BM) and diet changes in subjects (mean age= 60.1 ± 7.4 y; 55% female) with pre‐diabetes (fasting blood glucose 95–124 mg/dL on 2 occasions) participating in a 6‐mo. (6WL; N=29) or a 12‐mo. (12WL; N=28) weight loss (WL) intervention. 6WL included 150 min/wk supervised moderate aerobic exercise plus a WL program. 12WL was 6 mo. supervised exercise only, followed by 6 mo. WL intervention plus (unsupervised) exercise. BM was assessed at baseline (B) and at end of treatment (ET). Diet intakes (24‐hr recall, 3‐d record) collected at B and ET were analyzed for calories (kcal) and macronutrients. Between B and ET, BM was reduced (p<0.001) in both groups: −6.2 ± 7.7 kg (−6.7%) and −6.8 ± 5.4 kg (− 7.7 %) in 6WL and 12WL, respectively. Likewise, kcal intakes were reduced (p<0.0001): −462 ± 463 and −428 ± 459 kcal in 6WL and 12WL. Both groups increased %kcal protein (+2.1 ± 3.3 and +1.9 ± 3.8) and decreased %kcal fat (−4.7 ± 7.0 and −3.4 ± 6.3), kcal from saturated fat (−97 ± 98 and −76 ± 91) and total cholesterol (mg) (−82 ± 107 and −56 ± 135) intakes. Thus, subjects on an intensive diet and exercise intervention for 12 mo. were able to sustain both their WL and a number of other healthy dietary changes. Supported by NIDDK R01‐DK081559 and NIA AG000029.
Physical activity policies at childcare centers and impact on children's physical activity and screen‐time behaviors
Authors: Erinosho Temitope O , Hales Derek P , Vaughn Amber E , Mazzucca Stephanie , Ward Dianne S
Published: June 2021

This study evaluated the association between childcare center policies about physical activity (PA) and children's PA and screen‐time behaviors at 50 childcare centers. Data were collected in 2008–2009 using the Environment and Policy Assessment and Observation (EPAO) tool. Directors reported center PA policies (written, unwritten but general practice, no policy). Preschool children's PA and screen‐time behaviors were assessed via 4‐day observation at each center. Time spent per day in outdoor play, teacher–led PA, sedentary activities, and screen‐time were 67.7±33.6, 16.5±11.3, 52.2±28.8, and 39.7±47.9 minutes, respectively. Children engaged in more minutes of outdoor play at centers with a written policy about amount of active playtime; and a written/unwritten policy about amount of time spent outdoors. Written policies about amount of active playtime and time spent outdoors were also associated with more minutes of sedentary activity. Children engaged in more minutes of teacher‐led PA at centers with written policy about amount of time in teacher‐led activities. Children spent fewer minutes watching television and using computers at centers with written policies about such behaviors and written policy about staff supervision of media use. Results suggest that written and unwritten policies may be effective at increasing PA, but may be insufficient to help children achieve recommended levels of PA.
Reshaping breakfast: the smaller the cereal flake, the greater the intake
Authors: Rolls Barbara J. , Roe Liane S. , Meengs Jennifer S.
Published: June 2021

Cues related to food volume could influence both the portion taken and the amount eaten. This was tested by reducing the flake size of a breakfast cereal, so that as the flakes packed more closely together, a greater amount was needed to fill the bowl. In a crossover design, 40 adults ate breakfast in the laboratory once a week for four weeks. Across the meals, a standard cereal (Wheaties) was crushed to reduce the flake size and volume to 80%, 60%, or 40% of the standard. A constant weight of cereal was presented in an opaque container; subjects poured the amount they wanted into a bowl, added fat‐free milk and non‐caloric sweetener as desired, and consumed the meal ad libitum. Results showed that subjects poured a greater weight of cereal as the flake size decreased (p < 0.0001); the portion of the 40%‐size cereal averaged 1.5 times the weight of the standard cereal. Subjects ate about 97% of the cereal they took, thus total energy intake at the meal also increased as flake size decreased, from 289 ± 18 to 358 ± 20 kcal (p < 0.0001), a mean increase of 32 ± 7%. Despite this increase in intake, ratings of hunger and fullness did not vary across the meals. These findings show that perceptual cues related to food shape and volume can affect decisions about the portion of food taken, which in turn significantly influences energy intake. Supported by NIH grant R01DK059853.
Mealtime beverage and food intake to satiation interacts with meal advancement in healthy young men and women
Authors: El Khoury Dalia , Panahi Shirin , Luhovyy Bohdan L. , Goff H. Douglas , Anderson G. Harvey
Published: June 2021

Thirst, hunger, eating and drinking are closely interrelated. However, their interaction during meal progression and the impact of beverage type on this interrelationship have not been reported. In a randomized controlled study, 29 men and women (22 ± 0.4 years; 22 ± 0.3 kg/m2) consumed to satiation a pizza meal with one of water, 1% milk, regular cola, orange juice and diet cola. Mealtime food and fluid intake were measured within each of three 7‐min phases of the meal. A progressive decline occurred from phase 1 to 3 in fluid intake, averaging 59 ml, and food intake, averaging 268 kcal (P < 0.0001); however, the relative intake of fluid to food increased (P < 0.0001). Beverage type did not alter the overall association of fluid and food intake with meal progression. However, the effect of beverage type on fluid and food intake changed by phases of the meal. Although pizza intake was similar with all beverages averaging 932 kcal, the amount of fluid consumed was higher with orange juice than diet cola during phases 1 (P = 0.01) and 2 (P < 0.001) of the meal. Caloric beverages led to higher mealtime total energy intake compared to water (P < 0.001) and diet cola (P < 0.0001), during all three phases of the meal. Baseline thirst, but not appetite, correlated with fluid (r = 0.277; P = 0.001) and food (r = 0.163; P = 0.050) intake. In conclusion, meal progression was a major determinant of the relationship between mealtime beverage and food intake.

Supported by Natural Sciences and Engineering Research Council of Canada‐Collaborative Research and Development, Dairy Farmers of Ontario and Kraft Canada Inc.
Dietary supplement use is associated with positive mood states in US Military and Coast Guard personnel
Authors: Austin Krista Gail , McGraw Susan M , Lieberman Harris
Published: June 2021

Approximately 60% of armed forces personnel consume a dietary supplement (DS). Therefore, we investigated the association of mood with multiple classes of DS in military (M) and Coast Guard (CG) personnel (N = 5,536). Participants completed a survey of DS use and the Quick Mood Scale to assess wakeful‐drowsiness, relaxed‐anxious, cheerful‐depressed, friendly‐aggression, clearheaded‐confused and well coordinated‐clumsy mood domains. Supplements were categorized as multivitamin/mineral (MVM), individual vitamin/minerals (IVM), protein/amino acids (P), combination products (C), herbals (H), purported steroid analogs (S) and other (O). A one‐way analysis of covariance assessed effects of DS on mood controlling for age. Users of DS, especially MVM and P, reported being more awake (5.23 + 0.03 vs. 5.03 + 0.05), cheerful (5.48 + 0.03 vs. 5.25 + 0.04), friendly (5.58 + 0.03 vs. 5.44 + 0.04), clearheaded (6.01 + 0.03 vs. 5.74 + 0.04), and coordinated (6.02 + 0.03 vs. 5.77 + 0.04), (p<.01). Users of S felt less friendly (more aggressive) (4.79 + 0.18 vs. 5.55 + 0.02, p<.01). As no mechanisms can realistically explain the diverse effects of so many types of DS on different moods, we hypothesize this association is not causal, and DS intake per se does not alter mood. Studies of DS should consider whether mood differences in users vs. non‐users could be a confounding factor.

Support: US MRMC/DoD Center Alliance for DS Research
Micronutrient Intakes Vary by Age Group and Ethnicity in Older US Populations
Authors: Bird Julia K , Fulgoni Victor L
Published: June 2021

Older adults are at greater risk of malnutrition. Causes include reduced food intake, poorer taste and smell, difficulties chewing or preparing food, nutrient malabsorption in the aging digestive tract, and co‐morbidities. In addition, ethnicity may confound nutrient status. Using NHANES, 2003–2008, usual intakes of vitamins, minerals and carotenoids and percentage not meeting the EAR, where available, were determined using the National Cancer Institute method. Statistically significant differences between age groups (51–70, 71+ yrs) were calculated using a Student's t‐test and by linear regression modeling (significance was p<0.05). Defining at risk nutrients as more than 25% of population not meeting the EAR, adults 51+ yrs had lower intakes of at risk nutrients: calcium choline, lycopene, magnesium and vitamin E, and higher intakes of alpha‐ and beta‐carotene, vitamin A, C and D than adults aged 19–50 yrs. Significant differences were found comparing ages 51–70 and 71+ yrs, with lower intakes generally found in the older age group.

Ethnicity significantly affected adults’ nutrient intakes for most nutrients with African Americans having the lowest intakes of all nutrients except lutein+zeaxanthin, vitamins A, C and K. In summary, nutrient intakes vary by age group and ethnicity. Adults 71+ yrs and African American seniors are at higher nutritional risk.
25‐hydroxyvitamin D and adiposity in older black and white adults: the Health ABC study
Authors: Houston Denise K , Tooze Janet A , Hausman Dorothy B , Johnson Mary Ann , Cauley Jane A , Bauer Douglas C , Tylavsky Frances A , Harris Tamara B , Kritchevsky Stephen B
Published: June 2021

Lower 25‐hydroxyvitamin D (25[OH]D) with increasing adiposity has been attributed to decreased sun exposure because of limited mobility and increased vitamin D storage in adipose tissue. However, low 25(OH)D may also lead to increased adiposity via increased PTH, promoting calcium influx into adipocytes and enhancing lipogenesis. Associations between adiposity and 25(OH)D were examined in 2,369 older black and white adults in the Health ABC study (mean age, 74.6 yrs; 50.6% women, 38.3% black). Serum 25(OH)D was measured by DiaSorin RIA. BMI, waist circumference (WC), % body fat by DXA, and visceral (VAT) and subcutaneous (SAT) abdominal fat area by CT were measured at baseline and 4 years later. In whites, BMI (β±SE: − 0.38±0.06), % body fat (−0.20±0.05), WC (−0.11±0.02), VAT (− 0.022±0.004) and SAT (−0.011±0.003) were inversely associated with 25(OH)D at baseline after adjusting for age, gender, education, site, season, height, smoking, alcohol, physical activity, vitamin D intake and vitamin D supplement use (all p<0.0001). In blacks, only WC (−0.05±0.02, p=0.01) and VAT (−0.013±0.004, p=0.002) were associated with 25(OH)D at baseline. Baseline 25(OH)D was not associated with change in total or abdominal adiposity in whites or blacks. In summary, 25(OH)D was inversely associated with baseline adiposity in whites and blacks but not with change in adiposity. Supported by the NIA (R01 AG029364 & K01 AG030506).
Obesity affects short‐term folic acid pharmacokinetics in women of childbearing age
Authors: da Silva Vanessa R. , Hausman Dorothy B. , Kauwell Gail P. , Tackett Randall , Rathbun Stephen , Bailey Lynn B.
Published: June 2021

Maternal folate status and BMI are independent risk factors for neural tube defects (NTD). Epidemiological studies have identified an inverse association between serum folate and BMI, after adjusting for intake. The objective of this study was to compare the relationship between BMI and the short‐term pharmacokinetic response to an oral dose of folic acid. Healthy obese (BMI ≥ 30.0; n=16) and normal weight (BMI 18.4 – 24.9; n=16) women of childbearing age (18 – 35 y) were administered a single oral dose of folic acid (400 μg). Blood samples were collected over a 10‐h period to evaluate the serum folate response. Fasting baseline serum folate was lower in the obese group (p= 0.005); in contrast, red blood cell folate was higher (p=0.05). Area‐under‐the‐curve for the absorption phase (0–3 h) and peak serum folate were lower in obese vs normal weight women (p<0.005). Overall serum folate response (0–10 h) differed between obese and normal weight women (repeated‐measures ANOVA, p=0.001). Data suggest body distribution of folic acid is significantly affected by obesity and should pregnancy occur may reduce the amount of folate available to the developing embryo. These findings provide additional support for a BMI‐adjusted folic acid intake recommendation for NTD risk reduction.

Funded in part by HATCH #GEO00706 and #GEO00707
Transcobalamin I (TC I) deficiency, a common cause of falsely low serum cobalamin (Cbl), is usually genetic, and plasma corrinoid analogs are decreased: is “withholding” Cbl and its analogs from human cells and the microbiome the chief biological role for TC I?
Authors: Carmel Ralph , Parker James , Kelman Zvi
Published: June 2021

TC I is the most ubiquitous and abundant human Cbl‐binding protein but differs by not promoting Cbl uptake by cells. It carries all nonfunctional corrinoid analogs and >;70% of Cbl in plasma. Once thought rare, mild TC I deficiency (mTC I def) may cause 15% of low Cbl levels, which led us to examine TC I def and its implications for TC I biology. mTC I def and severe TC I def (sTC I def) were identified by TC I immunoassay. Plasma analogs were estimated by differential results between radioisotope dilution Cbl assays using TC I or intrinsic factor as binding proteins. All 4 studied patients with sTC I def were non‐Caucasian and homozygous or compound heterozygous for nonsense and deleterious missense TCN1 mutations (270delG; 315C>;T; 475T>;C). In mTC I def, 21 non‐Caucasian patients and affected relatives (14 families) were heterozygous for 270delG and 1 for 315C>;T, and 12 Caucasian patients (9 families) for 999G>;T, a deleterious missense mutation. Only 18 unrelated families with mTC I def had no detectable mutations. Analogs were decreased nonsignificantly in mTC I def plasma and absent in sTC I def. The TCN1 mutations in 24 of 42 families (57%) suggest that mTC I def is usually hereditary. We propose that the chief function of TC I is to withhold both Cbl and analogs from a variety of cells for a variety of reasons. The frequency of ethnically specific mutations suggests possible selective pressures to modulate TC I in diverse populations.
Maternal dietary Docosahexaenoic acid (DHA) supplementation prevents fetal growth restriction and pulmonary fibrosis caused by perinatal inflammation
Authors: Velten Markus , Britt Rodney D , Heyob Kathryn M , Tipple Trent E , Rogers Lynette K
Published: June 2021

Background

Epidemiological data suggest that maternal inflammation contributes to intrauterine growth restriction, preterm birth, and adult‐onset pulmonary diseases. DHA has anti‐inflammatory properties but responsible mechanisms are not defined.

Objective

We hypothesized that perinatal inflammation and oxidative stress induce a fibrotic phenotype by EMT induction and that this phenotype can be rescued by dietary DHA administration.

Methods

Pregnant C3H/HeN mice were injected on E 16 with LPS or saline and placed on control or DHA diets. Pups were exposed to 85% (O2) or RA for 2 weeks than returned to room air (RA). Protein, RNA, histological, and pulmonary function analyses were performed.

Results

LPS/O2 exposure reduced birth weight, decreased alveolarization, impaired pulmonary function, induced inflammation, matrix remodeling, EMT, and collagen deposition. Maternal DHA supplementation attenuated deleterious effects on birth weight, alveolarization, pulmonary mechanics, matrix remodeling, collagen deposition, and EMT in lung tissues of LPS/O2 mice.

Conclusions

Our data indicate that pulmonary morbidities at 8 weeks are due to maternal inflammation and neonatal oxidative stress. Perinatal DHA supplementation improves outcome by attenuating fibrotic response and improving alveolarization. These strategies would be straightforward to implement in pregnancy/lactation

Grant Funding Source: NIH: LKR R01AT006880, DFG: MV, VE 614/1–1
Genome‐wide transcriptome analysis of the effects of sulforphane on normal and prostate cancer cells
Authors: Beaver Laura Michelle , Buchanan Alex , Sokolowski Elizabeth I , Chang Jeff H , Wong Carmen P , Williams David E , Dashwood Roderick H , Ho Emily
Published: June 2021

Sulforaphane (SFN), a phytochemical derived from cruciferous vegetables, induces anti‐proliferative and pro‐apoptotic responses in prostate cancer cells, but not normal prostate cells. The mechanisms responsible for these specific chemopreventive properties remain unclear. We utilized RNA sequencing (RNA‐seq) to test the hypothesis that SFN modifies patterns of gene expression that are critical in prostate cancer progression. Normal prostate epithelial cells and prostate cancer cells were treated with 15 uM SFN and the transcriptome was determined at 6 and 24h time points. SFN alters the mRNA expression levels of ~3,000 genes in each sample and the transcriptional response is highly dynamic over time. SFN treatment also alters the expression of non‐coding RNAs including lincRNAs. SFN targets multiple pathways in normal and prostate cancer cells that are critical in cancer including proliferation, apoptosis, angiogenesis, DNA damage response, and cell migration. Network analysis suggests that the transcription factor Sp1 may be a major mediator by which SFN treatment induces changes in gene expression. Studies evaluating the effect of SFN on Sp1 are ongoing and Sp1 mRNA levels are significantly decreased by SFN treatment. Together, this information can be leveraged to help develop intervention strategies to significantly reduce the incidence and severity of prostate cancer. Funding: P01CA090890

Grant Funding Source: National Institute of Health
Glutaredoxin overexpression inhibits angiogenesis in vivo through regulation of soluble VEGFR1
Authors: Murdoch Colin E , Shuler Michaela M , Kikuchi Ryosuke , Walsh Kenneth , Cohen Richard A , Matsui Reiko
Published: June 2021

Glutaredoxin‐1 (Glrx) catalyzes the reduction of protein mixed disulfides (PSSG) and regulates function of proteins with redox‐sensitive thiols. We used Glrx transgenic (Tg) mice to investigate the role of Glrx overexpression on angiogenesis in vivo. Tg mice had impaired blood flow recovery and lower clinical function scores in an in vivo model of hind limb ischemia (HLI). VEGF‐induced aortic spouting and endothelial cell (EC) tube formation were attenuated in aortae and EC isolated from Tg mice. Interestingly, we observed an increase in soluble FLT (sFLT/sVEGFR1), the anti‐angiogenic VEGF receptor in Tg mouse plasma post‐HLI and media from Tg ECs. In addition, Glrx overexpression increased the non‐canonical Wnt ligand, Wnt5a, in both ischemic muscle and EC. Furthermore, Glrx induced‐sFLT1 expression in endothelial cells was attenuated by Box5, a competitive inhibitor of Wnt5a. Together these data suggest that Glrx overexpression negatively regulates angiogenesis post‐hind limb ischemia by regulating VEGF signaling through Wnt5a‐sFLT pathway. We speculate that enhanced Glrx expression may be critical to impaired angiogenesis observed in pathological conditions. (R37 HL104017 and PO1HL068758)
Role of Hedgehog in bleomycin induced pulmonary fibrosis
Authors: Hu Biao , Liu Jianhua , Wu Zhe , Liu Tianju , Ullenbruch Matthew R. , Phan Sem H.
Published: June 2021

Hedgehog signaling plays important roles in development and cell differentiation as well as in cancer. Recent studies indicate that it may be involved in stromal cell activation in the context of tumor desmoplastic reaction. In this study, the role of sonic hedgehog (SHH) in bleomycin induced pulmonary fibrosis is evaluated. Real time PCR analysis revealed that SHH was highly induced in alveolar type II cells but was not expressed by lung fibroblasts. However SHH treatment induced myofibroblast differentiation in fibroblasts. Gel shift analysis and site directed mutagenesis confirmed that a Gli1 binding consensus in the α‐smooth muscle actin promoter was important for mediating the SHH effect. Moreover repression of its downstream mediator Smo and Gli1 by shRNA, as well as by knocking out the Smo gene, prevented myofibroblast differentiation. Conditional mesenchymal cell specific knockout of Smo suppressed bleomycin induced pulmonary fibrosis. Thus, hedgehog signaling appears to be a key component of the fibrogenic response in this animal model.
RAGE signaling influences diesel particulate matter‐induced inflammation in primary alveolar macrophages
Authors: Barton David B , Bettridge Bryce C , Earley Tyler D , Robinson Adam B , Reynolds Paul R
Published: June 2021

Receptors for advanced glycation end‐products (RAGE) are members of the immunoglobulin superfamily of cell‐surface receptors implicated in pulmonary inflammation. In the present study, we tested the hypothesis that RAGE mediates inflammation in primary alveolar macrophages (AMs) triggered by diesel particulate matter (DPM). Using real time RT‐PCR, we discovered that RAGE was up‐regulated in primary AMs exposed to DPM for four hours. Because DPM increased RAGE expression, we exposed primary AMs isolated from RAGE null and wild type Balb/C mice to DPM. DPM led to the activation p38 and NF‐kB and that these two signaling intermediates were diminished in similarly exposed AMs isolated from RAGE null mice. Furthermore, TNF‐a, IL‐12, and IL‐13 were all significantly decreased in culture media from DPM‐exposed RAGE null AMs compared to similarly exposed WT AMs. These results demonstrate that primary AMs orchestrate diesel‐induced inflammation, at least in part, via RAGE‐mediated mechanisms. Further work may show that RAGE signaling is a potential target in treatment of inflammatory lung diseases exacerbated by environmental pollution. This work was supported by a grant from the Flight Attendant's Medical Research Institute (FAMRI, PRR) and a BYU Mentoring Environment Grant (PRR).
JAM‐A –mediated regulation of epithelial barrier function is dependent on a complex with ZO‐2, Afadin and PDZ‐GEF1 that modulates Rap2c activity.
Authors: Monteiro ana , Sumagin ronen , Rankin carl r , Leoni giovanna , Reiter dirk , Stehle thilo , Dermody terrence , Hall randy a , Nusrat asma , Parkos charles a
Published: June 2021

Intestinal mucosal barrier function is regulated by epithelial tight junctions (TJs), structures that control paracellular permeability. Junctional Adhesion Molecule A (JAM‐A) is a TJ‐associated protein known to regulate epithelial barrier, however mechanisms linking JAM‐A to regulation of permeability are incompletely understood. Proteomic, biochemical and cell‐biological approaches were used to identify candidate proteins involved in JAM‐A regulation of epithelial permeability. Our findings suggest that JAM‐A directly binds to the scaffold molecule ZO‐2 and indirectly associates with Afadin and PDZ‐GEF1 to regulate activation of the small GTPase Rap2c. A link to regulation of barrier was confirmed by siRNA‐mediated downregulation of these regulatory proteins leading to enhanced permeability. Since ZO‐2, Afadin and Rap2c have been implicated in cytoskeletal dynamics, we investigated whether JAM‐A regulation of barrier function is mediated through peri‐junctional actomyosin. While photobleaching experiments suggest that JAM‐A does not regulate actin turnover, JAM‐A deficient cells display enhanced activation of RhoA, a known inducer of actomyosin contraction. These findings suggest that JAM‐A regulates epithelial permeability through association with a multimolecular signaling complex that activates Rap2c, modulating contraction of the peri‐junctional actin cytoskeleton.
Claudin‐4 promotes epithelial repair and restoration of barrier function via interactions with the small GTPase Rap1
Authors: Sutherland Katherine , Bentley Trevor , LaFemina Michael , Frank James
Published: June 2021

Epithelial barrier function in the lung is dependent on tight junction (TJ) integrity. Claudin proteins largely determine paracellular transport properties and changes in claudin protein levels alter epithelial permeability and alveolar fluid clearance in the lung. Recent data also support potential roles for claudins in epithelial development, differentiation, migration, and survival. We have found that cldn4 is upregulated by an EGFR‐dependent mechanism in a variety of in vivo and in vitro models of lung injury and loss of cldn4 delays repair. Using a discovery proteomics approach, we have found that cldn4 associates with the small GTPase Rap1. We hypothesize that a novel interaction involving Rap1 and cldn4 modulates localized Rap1 activity to influence known functions such as migration and junction maturation. Transfection of the 16HBE cell line with dominant‐negative or constitutively active Rap1 followed by coimmunoprecipitation determined that the cldn4 interaction with rap1 is activation state dependent. These data highlight a potentially unique role of cldn4 in the repairing epithelium. Cldn4 may accelerate repair by affecting the localization of active Rap1, particularly to sites of new tight junction formation. Further investigation of this repair pathway may reveal molecular targets for the treatment of patients with lung disease. NIH/NHLBI RO1HL88440, HL88440‐S2, T32 HL007185
P38MAPK is involved in loss of keratinocyte cohesion induced by plakoglobin but not desmoplakin depletion
Authors: Dehner Carina , Spindler Volker , Waschke Jens
Published: June 2021

Desmosomes are patch‐like cell contact structures that are essential for the integrity of the epidermis by providing adhesive strength between keratinocytes. The core is organized of transmembrane proteins of the desmosomal cadherin family, which bind to their counterparts on the adjacent cell via their extracellular domain and are linked intracellularly to the keratin filament network by adapter proteins such as plakoglobin (Pg) and desmoplakin (Dsp). Since in the severe skin blistering disease pemphigus autoantibodies bind to desmosomal cadherins and induce loss of keratinocyte cohesion at least in part via p38MAPK activation and cause depletion of desmosomal components, we evaluated the roles of Pg and Dsp in p38MAPK‐dependent loss of cell adhesion. siRNA‐mediated silencing of either Pg or Dsp protein expression reduced cohesion of cultured human keratinocytes in dissociation assays. However, Pg but not Dsp silencing induced activation of p38MAPK. Accordingly, cell dissociation was blunted by p38MAPK inhibition under conditions of reduced Pg expression only. Interestingly, keratin filament retraction, a hallmark of pemphigus, was prominent under diminished Pg levels, an effect prevented by p38MAPK inhibition. Our data indicate that p38MAPK‐dependent keratin filament collapse is involved in loss of cell adhesion in response to Pg but not Dsp depletion.
The inflammatory cytokine IFNγ regulates intestinal epithelial homeostasis by controlling the spatiotemporal localization of Akt, 14.3.3ζ and β‐catenin.
Authors: Nava Porfirio , Kamekura Ryuta , Hamilton Ross , Medina‐Contreras Oscar , Kolegraff Keli N. , Denning Timothy L. , Parkos Charles , Nusrat Asma
Published: June 2021

The proinflammatory cytokine IFNγ modulates intestinal epithelial (IEC) homeostasis through effects on IEC proliferation. Epithelial proliferative responses are, in turn, regulated by β‐catenin, an adherens junction‐associated protein that differentially controls cell‐cell adhesion and gene transcription. Chronic exposure to IFNγ results in protein kinase B (PKB/Akt)‐mediated β‐catenin signaling that initially promotes, and subsequently suppresses IEC proliferation. The mechanism(s) governing the suppression of β‐catenin signaling are poorly understood. Here we show that the β‐catenin dependent, antiproliferative activity of IFNγ requires synthesis, activation and subcellular compartmentalization of 14.3.3ζ scaffold protein and the Akt isoform Akt1. 14.3.3ζ associates with phosphorylated β‐ catenin (pβ‐cat552) in IECs exposed to IFNγ that in turn promotes β‐catenin dependent‐Akt1 expression. Akt1 upregulation and its accumulation in the nucleus results in translocation of 14.3.3ζ and pβ‐cat552 from the nucleus to the cytosol thereby suppressing β‐ catenin transactivation and IEC proliferation. These results provide a mechanism by which IFNγ decreases active β‐catenin in the nucleus to suppress IEC proliferation during inflammation.
Med1 subunit of the mediator complex is phosphorylated by AMPK
Authors: Jia Yuzhi , Viswakarma Navin , Bai Liang , Vluggens Aurore , Sandra Sandhya , Misra Parimal , Lukas Thomas J , Borensztajn Jayme , Zhu Yi‐Jun , Thimmapaya Bayar , Reddy Janardan K
Published: June 2021

Mediator, a large multi‐subunit complex, plays a pivotal role in gene transcription by linking gene specific transcription factors with pre‐initiation complex and RNA polymerase II to promote transcription. In the liver the key subunit of the Mediator complex Med1 interacts with several nuclear receptors and transcription factors including GATA factor and CEBPβ to influence liver specific gene expression. Because ablation of Med1 results in embryonic lethality in the mouse, we have developed conditional knockout of liver Med1 to study the role of Mediator complex in liver regeneration, nuclear receptor signaling in hepatocyte proliferation and the development of liver cancer. We have obtained evidence that hepatocytes lacking Med1 do not regenerate following partial hepatectomy and fail to give rise to tumors when challenged with hepatic carcinogens. Adenovirally‐driven overexpression of Med1 in mouse liver stimulates hepatocyte DNA replication indicating that overexpression of Med1 alone is sufficient for liver cell proliferation. Importantly, we have discovered that the AMP‐activated protein kinase (AMPK), an important cellular energy sensor, phosphorylates Med1 in vitro and in liver and HeLa cells. AMPK inhibition by Compound C, decreases hepatocyte proliferation induced by Med1 overexpression. Our results suggest that Med1 phosphorylation by AMPK may be important in Med1 induced cell proliferation. Supported by DK097249.
Concurrent Inhibition of MET & EGFR in Regenerating mice liver following a partial hepatectomy. Effects on survival
Authors: PARANJPE SHIRISH , Bowen William C , Paranjpe Tejas , Luo Jianhua , Bhave Vishakha , Prosser Denise , Lokshin Anna , Michalopoulos George K
Published: June 2021

The HGF receptor MET and the EGF receptor (EGFR) are mitogenic receptor‐tyrosine kinases for hepatocytes. We report the results on liver regeneration in mouse following concurrent inhibition of kinase activity of MET & EGFR by using Gefitinib and JNJ 38877605 respectively. Mice were administered two doses of Gefitinib (300 mg/kg) and JNJ 38877605 (100mg/kg) by oral gavages. We also inhibited c‐met and EGFR by using shRNA mediated gene knock down. Western blot analyses and real time QRTPCR revealed significant inhibition at both mRNA and protein level in mice treated with shRNAs. In mice treated with chemical inhibitors both. pMet & pEGFR levels were significantly reduced compared to vehicle treated controls. A survival analyses following a JO‐2 antibody challenge in the treated and control mice by constructing survival plots indicated that pro survival effects of a PHx was abrogated in the inhibitor and shRNA plasmid treated mice compared to controls. Luminex based assays for cytokines on mouse plasma samples obtained at various time points from both chemical inhibitor treated and shRNA treated animals were carried out. An increased expression in TNF and its family members, Fas and HGF was evident at 10 hours post Phx in mice treated with inhibitors shRNAs. We are currently investigating the mechanisms and the pathways activated by combined inhibition of the two receptors that lead to hepatocyte death.
Impact of development and genetic variation on human hepatic CYP2B6 expression and activity.
Authors: Gaedigk Andrea , Twist Greyson P , Gaedigk Roger , Dai Hongying , Riffel Amanda K , Leeder J Steven , Pearce Robin E
Published: June 2021

CYP2B6 metabolizes many drugs that are used clinically in children and adults. To better understand the developmental expression and variability in CYP2B6 activity, we characterized a large panel of human liver samples (24 fetal, 141 pediatric, 36 adult) for RNA and protein expression and activity (microsomal buproprion hydroxylation). Genotyping included six CYP2B6 SNPs and one SNP each for P450 oxidoreductase and PPARα. RNA expression was correlated with immunoreactive protein, activity and age (r=0.29, 0.79 and 0.27, respectively). A piece‐wise regression indicated a strong correlation above a defined cut‐off point between activity and expressed protein (r=0.74), but no significant correlation when activity and protein expression were low. In fetal samples, low levels of CYP2B6 mRNA were detected, but little or no protein or activity were observed. Levels of RNA, protein and activity increased markedly after birth. No gender effects were observed. A trend was seen between CYP2B6 rs3745274 (associated with CYP2B6*6 and *9) and decreased RNA expression, but not for any of the other SNPs tested. This study confirms that CYP2B6 expression and activity develops over the first months of life. Although a large range of CYP2B6 activity (>;500‐fold) was observed in this cohort, CYP2B6 allelic variation does not appear to be a predictor of CYP2B6 activity.

Supported by a grant from the Tom Keaveny Foundation.
Transport by OATP1B1 and OATP1B3 enhances cytotoxicity of EGCG and certain substituted quercetins
Authors: Zhang Yuchen , Hays Amanda , Noblett Alexander , Thapa Mahendra , Hua Duy H. , Hagenbuch Bruno
Published: June 2021

Organic Anion Transporting Polypeptide (OATP) 1B1 and 1B3 are multispecific uptake transporters expressed in human hepatocytes. Certain tumors also express OATP1B3. Flavonoids like quercetin and the green tea catechins have been shown to have anticancer activity in cancer cell lines. In the present study we investigated to what extent several substituted quercetins (code name: QC) would affect transport mediated by OATP1B1 and OATP1B3. We measured uptake of the radiolabeled model substrates estradiol‐17β‐glucuronide, estrone‐3‐sulfate and dehydroepiandrosterone sulfate (DHEAS) in the absence and presence of the QC compounds in stable Chinese Hamster Ovary (CHO) cells expressing OATP1B1 or OATP1B3. Most of the QC compounds were inhibitors of OATP‐mediate uptake suggesting that they could also be substrates. QC6 was a stimulator of OATP1B3‐ mediated uptake of estradiol‐17β‐glucuronide by decreasing the apparent Km value. We could demonstrate with cytotoxicity assays that EGCG and most QC compounds preferentially killed OATP‐expressing CHO cells. EGCG and QC3 were among the most cytotoxic compounds and killed OATP1B3 expressing cells selectively. Thus, EGCG and QC compounds could be promising lead compounds for the development of novel anticancer drugs to target OATP1B3‐expressing cancers such as pancreas, prostate, colon or breast cancers.
From Genes To Proteins: Bringing hands‐on molecular biology activities into middle school classrooms to promote STEM education
Authors: Dutnall Robert N. , Banner Valentyna , Booker Aja , Vizzo Emily , Kane Christine
Published: June 2021

We have initiated an outreach partnership to promote STEM education for middle school students. The San Diego Global Vision Academy (SDGVA) is a recently established K‐6 school with a diverse student population, and a curriculum that includes a focus on STEM and critical and innovative thinking. In this first year of our partnership, SDGVA students will be introduced to the structure of DNA and how it directs the production of proteins. Our overall goals are for students to develop a greater understanding of (i) the processes that control their development, and the basis for inheritance of characteristics, (ii) the basis for diseases such as cancer, and the applications of genetic manipulation in therapy and biotechnology, and (iii) opportunities for careers in the STEM fields in college and beyond. Towards these goals, we have designed a series of classroom activities that include use of hands‐on materials in classroom lessons. Students will also perform inquiry‐based, laboratory‐style experimental investigations, including extracting DNA from different sources, and looking at fluorescent proteins made in bacteria, under the guidance of students from USD. We also plan for SDGVA students to visit USD to interact with faculty and students in the STEM fields. We will describe the progress of our project, its impact on students, and our plans for the future development of this partnership.
Performance 2012: Engaging the public with the physiology and pharmacology of sporting success
Authors: Lewis David Ian , Mills Dan , Haigh Charlotte
Published: June 2021

There is considerable public interest in sport, with a significant upsurge following major international competitions. The 2012 Olympic Games therefore provided an ideal opportunity to explore, with the public, biomedical aspects of sport. Our aim was to provide them with knowledge and understanding of exercise physiology and pharmacology, and promote debate on the ethical issues which may arise.

Three events were organised. The first, targeted at young people gave them the opportunity to act as scientists in a mock drug testing laboratory followed by a debate on the use of gene doping in sport. The second, for individuals who provide sporting opportunities for young people, was a facilitated debate in which their roles and responsibilities towards their athletes was explored. The final event was a two‐day interactive exhibition for the general public which combined “have a go” sporting activities such as wheelchair basketball with those which focused more on the science or ethics of performance enhancement e.g. an interactive mind‐map where the public could contribute their ideas on the harms and benefits of gene doping.

Performance 2012 successfully engaged the public with the physiology and pharmacology of performance enhancement in sport. All resources are available via the project website www.fbs.leeds.ac.uk/performance2012/index.php

The support of the Wellcome Trust is gratefully acknowledged.
Public outreach in high school science laboratories
Authors: Huang Jenq‐Kuen , Johnson Thomas , Albrecht Adam , Wen Lisa
Published: June 2021

In response to a campaign to promote K‐12 outreach activities launched in 2009 by the American Society for Biochemistry and Molecular Biology, Undergraduate affiliate Networking (ASBMBUAN), we (members of the ASBMB‐UAN, Western Illinois University Chapter) have started to plan and implement science outreach activities in local high school science laboratories. Our goal is to promote the quality of science education in rural area school districts near Western Illinois University. These rural area high schools have limited budget for chemicals and instruments and the turnover rate of the science teachers is relatively high. Resulting from that, students have very little opportunity for hands‐on experiments in their science laboratory. We have brought two science outreach activities to high school science classes near Macomb, IL. Each activity was designed to fit the school's laboratory setting and can be completed in 45 minutes. Labs cover “big ideas” like physical separation and then use modern biochemical techniques like size exclusion chromatography to support student learning. Here we present the outreach activities performed at the rural area high schools science classes in Western Illinois. Supported by ASBMB‐UAN and the Western Illinois University.
Recording and statistical analysis of early zebrafish developmental patterns using in vivo multiphoton microscopy
Authors: Peyriéras Nadine
Published: June 2021

We take the cell lineage tree of living organisms as the basis for the reconstruction of their spatio‐temporal multiscale dynamics. Indeed, embryonic development leading from the egg cell to the whole organism can be fully described by the spatio temporal deployment of the cell lineage tree annotated with quantitative parameters for cell membrane and nucleus shape. This data can be automatically extracted from in toto 3D+time imaging of the developing organism and allows answering most of the questions of classical embryology such as cell fate, presumptive organs clonal history, cell proliferation rates, contribution of cell division and its characteristic features to shaping tissues and organs. These questions can be answered by using the digital specimen corresponding to the validated phenomenological reconstruction of 3D+time image data sets. Exploring cohorts of individuals with different genetic and environmental conditions allows to integrate the cellular and molecular levels of organisation and to assess the basis for the robustness and variability of embryonic development. The phenomenological reconstruction of multimodal and multiscale 3D+time data is a first step toward the theoretical modelling and numerical simulation of morphogenetic processes. Virtual specimens derived from simulations are compared with digital ones reconstructed from the microscopy data to explore parameter spaces. This paradigm is used for investigating the early embryonic development of the teleostean Danio rerio.
Visualizing and perturbing the embryonic cardiovascular system with light
Authors: Jenkins Michael W. , Wang Yves T. , Peterson Lindsy M. , Gu Shi , Karunamuni Ganga H. , Ma Pei , Rollins Andrew M. , Watanabe Michiko
Published: June 2021

Biophysical forces impinging on cardiac cells and tissues are important signals needed for proper development of the cardiovascular system. Unfortunately, the role of these signals are not fully identified nor well understood due to a lack of proper tools to quantitatively assess the causal relationship between mechanical/electrical forces and phenotype. We developed a suite of optical imaging systems (optical coherence tomography, optical coherence microscopy and optical mapping) capable of quantifying morphologic features (e.g. endocardial cushion size), cardiac function (e.g. blood flow, conduction) and biophysical forces (e.g. shear stress, etc.) in the early stages of heart development. Also, we are developing a new method (optical pacing) to precisely perturb cardiac dynamics. These set of tools will enable us to investigate how altered function in the earliest stages of heart development potentially lead to congenital heart defects. Research Support: NIH grants HL083048, HL095717 HL, HL115373.
What we can learn from hagfish embryology
Authors: Kuratani Shigeru
Published: June 2021

The cyclostomes, or hagfishes and lampreys, represent the most basal lineage of vertebrates, providing a key to understand craniofacial evolution. While the monophyly of cyclostomes has been supported by recent molecular analyses, phenotypic traits of the hagfish, especially the lack of some vertebrate‐defining features, as well as endodermal origin of the adenohypophysis, have been interpreted as exhibiting more ancestral state than the rest of vertebrates. It is also true that adult craniofacial anatomy of this animal cannot be compared easily with that of the lamprey. To clarify this, the craniofacial developmental pattern of staged embryos of a Japanese inshore hagfish, Eptatretus burgeri, will be described to show that their adenohypophysis arises ectodermally as in other vertebrates, consistent with the molecular phylogenetic data. The above finding also allowed us to identify a pan‐cyclostome embryonic pattern, not shared by crown gnathostomes, and comparative analyses implied that many of the hagfish‐specific traits can be explained by changes secondarily introduced into the hagfish lineage. Since the craniofacial morphology of some basal gnathostomes retains some cyclostome‐like features, we conclude that the pan‐cyclostome pattern would likely reflect the ancestral program for the craniofacial development for the entire living vertebrates.
Sympathosensory sprouting in aged mice that over express nerve growth factor
Authors: Smithson Laura J , Kawaja Michael D
Published: June 2021

In this study, we investigated the effects of age on post‐ganglionic sympathetic plexuses formation in trigeminal ganglia of transgenic mice that have elevated levels of nerve growth factor (NGF) in cells expressing the glial fibrillary acidic protein promoter (GFAPpr). Our group has previously shown the presence of tyrosine hydroxylase (TH)‐immunopositive plexuses around a subset of sensory somata of 2–3 month old GFAPpr‐NGF mice. This sympathosensory sprouting has been linked to the onset of neuropathic pain. In this current study, we demonstrated morphological changes of TH‐immunopositive plexuses in ganglia of aged (i.e., 11–14 month and 16–18 month old) GFAPpr‐NGF mice. Double immunolabeling revealed an association between reactive satellite glial cells (and some reactive macrophages) and TH‐immunopositive plexuses in aged mice, compared to younger transgenic mice. In addition, TH‐immunopositive plexuses in aged mice were associated with injured sensory neurons. Our results are the first to report changes in the morphology of sympathetic plexuses in sensory ganglia of aged mice. These results suggest that the presence of sympathetic plexuses around sensory somata may be slowly damaging the associated sensory neurons. That is, sympathosensory sprouting in aged mice likely compromises the function of sensory neurons, which may have profound implications for neuropathic pain. This work was supported by grants from Canadian Institutes of Health Research (CIHR; MDK) and by a Banting and Best Studentship from CIHR (LJS).