bluuebunny/crossref_metadata_2025 huggingface dataset:
Leptin increases adipose triglyceride lipase in bovine primary adipocytes
Authors: Koltes Dawn , Spurlock Diane
Published: June 2021

Proper leptin signaling is critical for energy homeostasis. Although, leptin induces a small but significant increase in lipolysis, mechanisms leading to leptin mediated lipolysis remain unclear. Our objective was to test the hypothesis that leptin alters adipose triglyceride lipase (ATGL) abundance via signal transducer and activator of transcription 3 (STAT3) signaling to regulate lipolysis. Bovine adipocytes were exposed to 0, 10, 100 or 200 ng/mL of bovine leptin for 2 hours to characterize the lipolytic response. Glycerol concentration in the media increased with 100 ng/mL of leptin compared to control adipocytes (P<0.02), but free fatty acid concentrations were similar between control and treated adipocytes (P>;0.80). Total protein abundance and phosphorylation of hormone sensitive lipase and perilipin were similar between control and treated adipocytes (P>;0.05). However, ATGL protein increased with 100 and 200 ng/mL of leptin (P<0.01). Bovine adipocytes were then pre‐treated with 0, 1, 1.25, 1.5, 1.75, or 2 μM of a STAT3 inhibitor, STATTIC, prior to treatment with 100 ng/ml of bovine leptin. Glycerol concentration and ATGL abundance were reduced to control levels with STATTIC concentrations greater than 1 μM. Results from this study suggest that leptin regulates lipolysis via a STAT3 mediated increase in ATGL abundance. Supported by USDA‐CSREES, NRI project 2009–35206–05222.
RNAseq Analysis of FFPE Radical Prostatectomy Specimens Identifies Predictors of Biochemical Recurrence
Authors: Moreno Carlos Sanchez , Long Qi , Sannigrahi Soma , Xu Jianpeng , Johnson Brent A , Zhou Wei , Nam Robert K , Seth Arun , Gillespie Theresa W , Osunkoya Adeboye O , Petros John A
Published: June 2021

To identify effective predictors of recurrence following radical prostatectomy, RNAseq libraries were prepared from RNA derived from 71 Formalin Fixed Paraffin Embedded (FFPE) prostatectomy samples from two institutions, and high‐throughput sequencing was performed on an Illumina HiSeq2000 instrument. Subjects were divided into high and low risk groups based on the optimal predictive score cutoff from ROC analysis, and we performed log rank test to compare biochemical recurrence (BCR) between the two risk groups. For the 70 samples analyzed, clinical variables alone were highly predictive of recurrence (log‐rank test p=2.55e‐04); however the panel of 11 genes from the RNAseq analysis outperformed clinical variables alone (log rank test p=1.82e‐13). Prediction using this model achieved an ROC AUC of 0.959 with 85% sensitivity and 97% specificity. This set of RNA biomarkers may be useful in the prediction of BCR following radical prostatectomy, irrespective of stage, grade, race, or pre‐operative PSA. Patients with tumors having increased expression of these markers may benefit from close follow‐up after surgery. Research supported by DOD IDEA Award W81XWH‐010–1‐0090.
Evo‐Devo of Specialized Bone Joints Formed During Pre‐Natal and Post‐Natal Ontogeny in Turtles With Shell‐Closing Systems
Authors: Cordero Gerardo Antonio , Stearns Samantha , Janzen Fredric
Published: June 2021

The evolution of complex skeletal traits is often the result of functional variation in developmental genes during pre‐natal life stages. However, in some vertebrates, the development of such traits extends into post‐natal ontogeny, during which the relationship between functional genetic variation and phenotypic change is less clear. The objective of this study was to characterize pre‐natal and post‐natal development of a complex skeletal trait in turtles. We focused on phylogenetic comparisons of turtles with shell‐closing systems, an adaptation that enables some turtles to fully conceal appendages during predator attacks. We combined classical embryological approaches with modern geometric morphometrics to describe and quantify developmental change in bone tissues of deeply divergent lineages. We validated previous observations on the embryonic development of a specialized bone joint that articulates the dorsal scapula with the ventral shell. This joint is critical for the function of shell‐closing systems and forms via secondary subdivision of cartilage tissue. We then quantified a dramatic anatomical transformation associated with the post‐natal development of an external bone joint on the ventral shell of a lineage with the most derived form of the shell‐closing phenotype. This joint was not present in neonate turtles but likely developed in sub‐adult/adult age classes via tissue remodeling that was mechanically induced (due to muscle abduction). Remarkably, the turtle shell‐closing system is a complex skeletal trait that could feature both genetically‐ and epigenetically‐activated developmental programs. We are currently studying the genetic basis of this phenotype but will explore options to test hypotheses on epigenetic mechanisms.

Grant Funding Source: NSF
Toward a better understanding of canonical and non‐canonical cap‐binding complex subunits of Arabidopsis thaliana
Authors: Patrick Ryan M , Mayberry Laura K , Choy Grace , Gebhard Mark C , Browning Karen S
Published: June 2021

The eIF4F cap‐binding complex, made up of the mRNA cap‐binding protein eIF4E and the large scaffolding protein eIF4G, is a key regulatory point for translational control in eukaryotes. Gene duplications of eIF4F proteins are common and may have evolved specialized functions. Higher plants have a conserved eIF4F complex as well as a plant‐specific eIFiso4F complex. We have used the increasing availability of plant genomic sequence data to trace the origin of the different cap‐binding complex constituents of the model plant Arabidopsis thaliana. Our findings indicate that the eIF4F complex and eIFiso4G are conserved throughout land plants and green algae, while eIFiso4E evolved recently after the emergence of flowering plants. We have also identified structural motifs that are signatures of eIF4G and eIFiso4G proteins. The Brassicaceae family, of which Arabidopsis is a member, has a unique form of eIF4E which we have found has divergent binding affinity for eIF4G in vitro but retains some translation‐enhancing function. These findings have helped us better define the canonical constituents of translation initiation in higher plants as a basis for continuing research. This research was supported by National Science Foundation grants to K.S.B. (MCB1052530 and MCB0745146)
Soluble epoxide hydrolase (sEH)‐ and UDP‐glucuronosyltransferase (UGT)‐dependent hypertension in pregnancy
Authors: Kim Hyesook , Joiakim Aby , Park Jung‐A , Kaplan David J , Taylor Robert N , Putt David
Published: June 2021

Approximately 5–8% of pregnant women have preeclampsia. Epoxyeicosatrienoic acids (EETs) generated by cytochromes (CYP) 2C and 2J have anti‐hypertensive activity. Urinary 14,15‐dihydroxyeicosatrienoic acid (DHET) levels in spontaneously hypertensive rats (SHR) were found to be ~35‐fold higher than levels in normotensive Wistar Kyoto (WKY) rats, suggesting that soluble epoxide hydrolase (sEH) induced developmental hypertension. Urinary levels of 14,15‐DHET were measured by ELISA in 7 preeclamptic and 9 healthy pregnant women. Mean systolic/diastolic pressures of the preeclamptic and healthy women were 152/95 and 120/74 mm Hg, respectively (p<0.01 for both). Contrary to the rat study, urinary free 14,15‐DHET levels did not significantly differ among the pregnant women. However, glucuronidated 14,15‐DHET levels in preeclamptic women was significantly higher (p<0.03) than controls. Glucuronidation of 14,15‐DHET (74 μM) by UDP‐glucuronosyltransferase (UGT) in pooled human liver microsomes was inhibited by 87% after treatment with 1.3 μM 12‐(3‐adamantan‐1‐yl‐ureido)dodecanoic acid (AUDA). The results demonstrated that increased urinary glucuronidated 14,15‐DHET is a biomarker for sEH‐ and UGT‐dependent preeclampsia and that AUDA inhibits both sEH and UGT, putative target enzymes for medical intervention. Supported in part by NIEHS SBIR Phase I (N43ES85430) and Phase II (N44ES05459).
ProPeL: A method to discover specific kinase motifs and predict target substrates
Authors: Chou Michael F , Prisic Sladjana , Lubner Joshua M , Church George M , Husson Robert N , Schwartz Daniel
Published: June 2021

Identification of kinase substrates is essential to understanding the role kinases play in normal and disease states. Kinases discriminate between potential substrates in part by recognizing linear sequence “motifs” of amino acids surrounding the phosphorylated residue. Current techniques for discovering kinase motifs are often expensive and labor intensive. The Proteomic Peptide Library (ProPeL) method is a novel approach to discover kinase specificity motifs de novo. Briefly, human kinases are expressed in E. coli and allowed to phosphorylate bacterial proteins. After enzymatic digestion of the bacterial lysate, tandem mass spectrometry is performed and motifs are extracted from the identified peptide sequences using the authors’ pLogo and motif‐x software. The ProPeL methodology successfully recapitulated the known and diverse motifs for human basophilic Protein Kinase A (PKA) and acidophilic Casein Kinase II (CK II). We then used the motifs determined using the ProPeL methodology to directly predict with high accuracy, potential target substrates of each kinase in the human proteome using the authors’ scan‐x program. We thus demonstrate the validity of the found motifs and the approach to search for potential substrates. This study was funded in part by grants from the University of Connecticut Research Foundation (DS) and Genomes to Life from US Department of Energy (GMC).
Evaluating the relationship between fruit and vegetable intake using plasma and dermal biomarkers and reported dietary intake in 4th grade children
Authors: Nguyen Lori , Scherr R E , Linnell J , Nicholson Y , Spezzano T , Brian K , Briggs M , Ermakov I , Gellermann W , Keen C , Miyamoto S , Smith M , Steinberg F , Young H M , Zidenberg‐Cherr S
Published: June 2021

The objective of this study is to evaluate the relationship between biomarkers of fruit and vegetable intake comparing plasma and dermal carotenoids to reported intake. It is important to have a valid, objective, and non‐invasive method to assess nutritional status in children and a feasible data collection method. As part of the multi‐component nutrition education intervention Shaping Healthy Choices, a health fair was hosted to collect biochemical data from 4th graders. Dermal carotenoids were measured using Resonance Raman Spectroscopy (RRS). Plasma carotenoids were quantified using High Performance Liquid Chromatography. Dietary intake was assessed using the Block Food Frequency Questionnaire. Total plasma carotenoids strongly correlated with RRS values when corrected for BMI (R2=0.80; P<0.01; n=10). RRS values strongly correlated with FFQ orange and leafy vegetable servings (R2=0.80; P<0.03 and R2=0.83; P<0.02, respectively). This study shows that a health fair is a feasible model to collect valid biomarker data assessing fruit and vegetable intake. RRS is a valid, non‐invasive method for determining carotenoids and correlates with reported intake. This validation work serves as justification for larger sample sizes in future studies. This model will be used to gather biomarker data at postevaluation and in future nutrition intervention studies.

Grant Funding Source: UCANR and USDA 2011–38420–20082
The development and implementation of an inquiry‐based nutrition education curriculum for the Shaping Healthy Choices Program
Authors: Linnell Jessica , Scherr Rachel , Nicholson Yvonne , Spezzano Theresa , Brian Kelley , Zidenberg‐Cherr Sheri , Smith Martin H.
Published: June 2021

The objective of this study was to develop and evaluate an inquiry‐based nutrition education curriculum that includes garden‐based extension activities. The curriculum is intended to help students learn key nutrition concepts and improve science process skills that are important for making healthy choices. Curriculum topics include: roles of nutrients, MyPlate recommendations, physical activity, reading food labels, understanding consumerism, and making healthy snacks. This curriculum was implemented among 4th‐grade children as part of the Shaping Healthy Choices Program (SHCP), a multi‐component, controlled intervention. Schools meeting inclusion criteria were randomly assigned to control (C) or intervention (I) groups. Participation was n=133 (C) and n=177 (I). Percent eligibility in the Free and Reduced‐Price Meal Program was 30–35%. Basic Test of Science Processing Skills (BAPS) mean scores were 25.9 (C) and 25.6 (I) out of 35, and nutrition knowledge mean scores were 20.2 (C) and 19.5 (I) out of 35. BAPS was correlated with nutrition knowledge (r=.48; p<.001). Qualitative data included student use of science process skills in nutrition‐related problems and journal reflections from classroom teachers. Teachers reported enthusiasm by students for lesson content and student learning exceeded their expectations. Supported by UCANR and USDA 2011–38420‐20082.
The Shaping Healthy Choices Program: A Multi‐Component, School‐Based Approach to Improve Children's Nutrition and Health Behaviors While Supporting Regional Agriculture
Authors: Scherr Rachel E , Linnell J , Smith M , Nicholson Y , Spezzano T , Bergman J , Brian K , Briggs M , Feenstra G , Hillhouse C , Keen C L , Nguyen L , Ontai L , Schaefer S , Steinberg F , Sutter C , Wright J , Young H , Zidenberg‐Cherr S
Published: June 2021

The Shaping Healthy Choices Program (SHCP) involves the integration of five components: nutrition education; family and community partnerships; regional agriculture; foods available on school site; and school wellness policies. This study uses a pre‐/post‐test design to test the hypothesis that schools utilizing the SHCP will improve nutrition knowledge, science processing skills, and health‐related behaviors. Schools meeting inclusion criteria were randomly assigned to control (C) or intervention (I) groups. All 4th grade students were invited to participate with a 98% (n=133) and 97% (n=177) rate of participation in C and I schools, respectively. The ethnic diversity index was 63 and 75, and free and reduced‐price eligibility was 30% and 35%, at C and I, respectively. The cornerstone of the SHCP was 20 h of inquiry‐based, garden‐enhanced nutrition education with accompanying parent newsletters. An increased variety of regional produce was available at I schools. The BMI for C and I students was 18.7 and 20.7, respectively. Mean reported MyPlate vegetable servings for C and I were .72 and .57, respectively. Mean student vegetable preference scores (max=5) for C and I were 3.5 and 3.6, respectively. This model is useful for data collection and evaluation of multi‐component school‐based nutrition interventions. Supported by UCANR and USDA 2011–38420‐20082.
Friends Matter: Food safety cognitive and behavioral influences among youth
Authors: Quick Virginia , Heller Rebecca , Corda Kirsten , Martin‐Biggers Jennifer , Byrd‐Bredbenner Carol
Published: June 2021

Do perceptions of food safety cognitions and behaviors of friends differ from one's own perceptions? This study examined youths’ food safety cognitions and behaviors and perceptions of their friends’ food safety cognitions and behaviors. Data were collected using an online survey based on Health Belief Model, Theory of Reasoned Action, and social norms theory constructs. Youth (n=227; mean age 12.8±0.7y; 59% female) were recruited from 4 middle/high schools in 4 states. Paired t‐tests revealed significant differences (P<0.001); compared to themselves, youth felt that their friends perceived themselves to be less susceptible to food poisoning, were less likely to handle food safely, had more negative food safety attitudes, and had less food safety self‐efficacy. Findings suggest that youth believe they have more positive food safety cognitions and behaviors and have greater recognition of their susceptibility to food poisoning than their peers do. Findings suggest that food safety behavior change interventions should consider coupling perceptions of friends’ food safety cognitions and behaviors with concern for friends as a method for leveraging positive peer pressure to promote food safety behavior change in their social networks.

Grant Funding Source: USDA, National Food Safety Initiative and in part by the intramural research program of the National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development.
Dietary Supplementation with L‐Arginine between Days 14 and 25 of Gestation Enhances Litter Size in Gilts
Authors: Li Xilong , Bazer Fuller , Johnson Gregory , Burghardt Robert , Frank James , Dai Zhaolai , Wang Junjun , Shinzato Izuru , Wu Guoyao
Published: June 2021

Gilts were checked daily for estrus with boars in the morning and bred at onset of the second estrus and 12 h later (the time of breeding = d 0 of gestation). Between d 14 and 25 of gestation, 15 gilts per treatment were housed individually and fed twice daily 1 kg of a corn‐and soybean meal‐based diet supplemented with 0.0%, 0.4%, or 0.8% L‐arginine. All diets were made isonitrogenous by addition of L‐alanine. At d 25 of gestation, gilts were hysterectomized to obtain conceptuses. Compared with control gilts, dietary supplementation with 0.4% or 0.8% L‐arginine increased (P ≤ 0.05) concentrations of arginine in maternal plasma, total volume of amniotic fluid, total amounts of arginine in allantoic and amniotic fluids, total amounts of fructose and most amino acids in amniotic fluid, placental growth, and the number of viable fetuses per litter by 2. The numbers of total fetuses or corpora lutea, total fetal weight, total volume of allantoic fluid, maternal circulating levels of progesterone and estrogen, or total amounts of the hormones in allantoic fluid did not differ among the three treatment groups. Reproductive performance of gilts did not differ between the 0.4% and 0.8% L‐arginine groups. These results indicate that dietary supplementation with 0.4% or 0.8% L‐arginine between d 14 and 25 of gestation can enhance embryonic/fetal survival in gilts. Supported by USDA grants.
Free Amino Acid Content of Equine Muscle Changes in Response to Graded Levels of Lysine Intake
Authors: Tanner Sara , Good Lindsay , DeLuca Elizabeth , Coleman Robert , Urschel Kristine
Published: June 2021

In typical equine diets lysine is the first limiting amino acid (AA) making it of particular interest in growing horses. Six thoroughbred colts (401±5 d) were used to study six levels of lysine intake: 76, 90, 104, 118, 127, & 136 mg/kg/d. Diets were isonitrogenous and isocaloric. Each horse was fed each diet in a random order for 6 d utilizing a 6×6 Latin square design. On d 6 venipuncture samples were taken into heparinized vacutainers before the concentrate portion of the feeding and 90 min post‐feeding. Muscle biopsies were taken ~100 min post‐feeding from the gluteus medius and flash frozen. Muscle samples were freeze dried and homogenized. Plasma and homogenized muscle were analyzed via HPLC. Plasma lysine concentrations increased in a dose dependent manner with increasing lysine intake (P<0.0001). Muscle AAs had different response patterns. Taurine concentrations were greatest for horses receiving the greatest lysine intake (P=0.01). Arginine (P=0.03) and ornithine (P=0.10) concentrations were greatest in the horses receiving the highest and the lowest levels of lysine intake. Lysine tended (P=0.10) to respond, at first decreasing and then increasing with lysine intake. These results indicate that dietary lysine intake affects AAs that are structurally related to lysine, namely ornithine and arginine, and the mechanisms governing this effect require additional investigation. This project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2010–65206‐20638 from the USDA National Institute of Food and Agriculture.
Pica behavior is prevalent and associated with low iron status in pregnant adolescents
Authors: Lumish Rachel Allison , Young Sera L , Lee Sunmin , Cooper Elizabeth M , Pressman Eva , O'Brien Kimberly O
Published: June 2021

A relationship between iron deficiency, pregnancy, and pica (the craving and purposive consumption of non‐food items) has been frequently observed. However, few studies have related pica behaviors to biomarkers of iron status, and little is known about pica prevalence in pregnant adolescents. To address this, we undertook a longitudinal study examining iron status and pica behaviors among a group of 158 pregnant adolescents (≤ 18 y). Approximately two‐thirds of the participants were African American and 25% were Hispanic. Maternal iron status [hemoglobin (Hb), transferrin receptor (TfR), serum ferritin (SF), total body iron (TBI), and hepcidin (Hep)] was assessed at mid‐gestation (~26 weeks) and delivery. Pica and other characteristics were assessed up to three times across gestation. Among the 158 adolescents, 45% reported engaging in pica at some point during pregnancy. Substances ingested included ice, starches, powder, and plastic. At mid‐gestation, mean SF, TBI, and Hep values in the pica group were significantly lower (p<0.05) than those in the non‐pica group. The prevalence of anemia (Hb≤10.5 g/dL) at mid‐gestation was significantly higher (p<0.002) in the pica group (27.8%) than among those who did not report pica (6.5%). While further studies must address the etiology of these relationships, our findings suggest that pica could be an inexpensive method for screening for iron deficiency.

Grant Funding Source: USDA 2005–35200 and 2008–01857
Repeated dosing and BMI influence plasma polyphenol response in humans
Authors: Novotny Janet A. , Chen Tzu‐Ying , Terekhov Anton I. , Baer David J. , Gebauer Sarah K. , Ho Lap , Wang Jun , Pasinetti Giulio M. , Ferruzzi Mario G.
Published: June 2021

Repeated dosing of grape polyphenols has been found to affect bioavailability and metabolism of catechin (C) and epicatechin (EC) in rats. In addition, obesity may influence intestinal function and xenobiotic metabolism, which could alter polyphenol bioavailability. To assess the impact of exposure and obesity on polyphenol bioavailability in humans, a pilot 11 day intervention was conducted with 12 adults with either normal BMI (23.5±0.7) or high BMI (30.6±0.6). Volunteers consumed 250 mg resveratrol, 900 mg grape seed extract, and 20 oz. Concord grape juice daily for 11 d. On d1 and d11, treatment was consumed after a 12h fast, then serial blood samples were collected for 6h. Plasma was extracted, and deconjugated C, EC, resveratrol (R), and quercetin (Q) were measured by LC‐MS/MS. Plasma AUCs were greater for C, EC, and Q on d11 compared to d1 for low BMI (P<0.039) but not high BMI individuals. Further, AUCs were greater for EC and R for low BMI compared to high BMI individuals (P<0.041), with a similar trend for C (p=0.065), on d11 but not d1. These results suggest that that repeated exposure and obesity likely influences polyphenol bioavailability in humans. Supported by the USDA and NCCAM.
Acetyl‐CoA carboxylases are checkpoints in adipocyte differentiation
Authors: Cordonier Elizabeth Lyn , Teixeira Daniel Camara , Han Zhonghi , Pannier Angela K , Zempleni Janos
Published: June 2021

Acetyl‐CoA carboxylases (ACC) 1 and 2 depend on biotin as a coenzyme and catalyze the carboxylation of acetyl‐CoA to malonyl‐CoA. Malonyl‐CoA produced by cytoplasmic ACC1 and mitochondrial ACC2 is a precursor in fatty acid (FA) synthesis and an inhibitor of mitochondrial FA oxidation, respectively. We hypothesized that ACCs are checkpoints in adipocyte differentiation, and used human mesenchymal stem cells (hMSC) and murine 3T3‐L1 preadipocytes to test our hypothesis. The expression of ACC2 increased by 1500% in differentiating hMSC compared with non‐differentiating hMSC, judged by qRT‐PCR and western blot. This increase preceded the increase of adipocyte marker genes FABP4 and PPARy. Treatment of hMSCs with grape leaf extract (GLE) inhibited the differentiation into adipocytes judged by the abundance of PPARy/FABP4 mRNA and staining of lipid droplets with Oil Red‐O. Likewise, treatment of 3T3‐L1 cells with the microbial ACC inhibitor soraphen A inhibited differentiation, judged by decreased lipid accumulation. Treatment of transgenic fruit flies, predisposed to storing excess body lipids, with GLE decreased body lipids by ~50%. In future studies, we will use mutagenesis to determine which of the two ACCs is the critical checkpoint in differentiation. We conclude that ACCs are checkpoints in adipocyte differentiation and that manipulation of ACC activity decreases body fat.

Grant Funding Source: ARD Hatch, NIFA, and NIH
The effect of calorie restriction with and without curcumin supplementation on inflammatory and T cell‐mediated immune responses of obese mice
Authors: Wang Junpeng , Vanegas Sally M , Du Xiaogang , Noble Timothy , Zingg Jean‐Marc A , Meydani Mohsen , Meydani Simin Nikbin , Wu Dayong
Published: June 2021

Obesity is associated with low‐grade inflammation and impaired T cell‐mediated immune response. Caloric restriction (CR) is shown to reduce inflammatory response and enhance cell‐mediated immune function. Spice compound curcumin (C) is proposed to have anti‐obesity and anti‐inflammation properties while piperine (P) enhances its bioavailability and efficacy. We determined if C/P could potentiate CR's beneficial effect on immune and inflammatory responses in diet‐induced obesity. Mice were fed a high‐fat diet (HFD) for 22‐wk followed by a CR diet (10% for 5‐wk and then 20% for additional 33‐wk), in the presence or absence of C and/or P. Results showed that compared to HFD control, all CR mice, with or without C and/or P, had fewer total spleen cells but a higher percentage of CD4+ T cells. CR resulted in a lower production of pro‐inflammatory cytokines IL‐1β and TNFα, and prostaglandin E2, as well as a trend toward lower IL‐6. Mice with CR alone had higher T cell proliferation and IL‐2 production, but this effect of CR was blunted by C and/or P. CR alone or in combination with C and/or P had no effect on production of cytokines IL‐4, IL‐10, IFN‐γ, TGF‐β, and IL‐17, or the proportion of different CD4+ T cell subsets. In conclusion, CR reduces inflammatory response and enhances T cell‐mediated function in HFD fed mice, but the presence of C and/or P does not provide additional benefit. Supported by the USDA contract #58–1950‐0–014.
Modulating dietary vitamin A altered the pathogenicity of Citrobacter rodentium in mice
Authors: Smith Allen D , Cheung Lumei
Published: June 2021

Vitamin A deficiency (VAD) is associated with increased bacterial infection‐related morbidity and mortality. Citrobacter rodentium (Cr) is a mouse pathogen that mimics many aspects of pathogenic E. coli infections in humans and is used to study mucosal immunity to enteric bacteria. Mice fed VA sufficient (VAS)‐ and VAD‐diets were infected with Cr and groups of infected VAD‐mice were switched to VAS‐diet on day 0 (D0) or seven (D7) post‐infection. Fecal excretion of Cr was monitored and bacterial load in the colon and spleen assessed at necropsy (day 12). The VAD‐mice infected with Cr had higher mortality than VAS‐or VAD‐mice switched to a VAS diet. The VAD‐mice had 3–4 log10 higher levels of Cr in the colon than VAS‐mice and bacteria were recovered from spleens of VAD‐ but not VAS‐mice indicating altered intestinal barrier function. Infected VAD‐mice switched to a VAS diet at D0 or D7 had colonic Cr levels between those of VAD‐ and VAS‐mice, and less bacteria was recovered from their spleens indicating partial restoration of barrier function. Growth of Cr was inhibited by D21 in mice on all diets except VAD‐mice which also had decreased mRNA expression of IFN‐γ and IL12p40 in the colon that correlated with increased Cr levels. In conclusion, the severity of Cr infection in mice is modulated by dietary levels of VA and supports VA's role in mucosal immunity to an enteric pathogen. Funded by USDA/ARS Project Plan 1235–51000‐055–00D.
Adolescent traumatic stress experience shapes adulthood fear conditioning and extinction responses
Authors: Moore Nicole LT , Gauchan Sangeeta , Genovese Raymond F
Published: June 2021

A traumatic stress paradigm (underwater trauma, UWT) was used to model an intense developmental stress event during adolescence, and evaluate effects on fear and stress response lasting into adulthood. Adolescent rats (P37) were exposed to UWT or swim without submersion. Rats were then allowed to reach adulthood (P114) and fear conditioned using light and sound (conditioned stimulus), with or without footshock (unconditioned stimulus). After a three‐week fear incubation period, rats were given one extinction trial per day for five consecutive days using an operant behavior conditioned suppression task. Preliminary results suggest that adolescent traumatic stress experience shapes some aspects of adulthood stress response severity, and residual fear responses after five extinction trials.

This work is supported by the Military Operational Medicine Research Program, US Army Medical Research and Materiel Command.
Novel model of thermoregulation based on control theory used to evaluate peripheral microvascular function
Authors: Chekh Viktor , Soliz Peter , Barriga Simon , McGrew Elizabeth , Kanagy Nancy , Luan Shuang
Published: June 2021

A novel heat transfer model has been developed to describe the thermal interaction of human tissue with the external environment. Thermal regulation is explicitly represented using feedback mechanism from control theory. The model was applied to data obtained from measuring the temperature of regions of interest (ROI) on the surface of the plantar foot with an infrared camera before and after a cold pressor test. The parameters of the model have been computed from the experimental results using numerical optimization. The robustness has been proven by interpolation of incomplete data: the model accurately (within 0.05–0.3□C) predicts the subject's recovery temperature. In addition, the results based on 20 subjects (7 normal (N), 7 diabetic (D) and 6 diabetic with diagnosed peripheral neuropathy (PN)) clearly show that the quality of body heat recovery, as well as the speed of thermoregulation are impaired by damage to the peripheral nerves and secondary vasculature dysfunction, suggest that this method could be efficiently used for early diagnosis of diabetic peripheral neuropathy. Preliminary results from one model parameter show that the average recovery rate after one minute for certain ROI was 5–30% for N, about 0% for D, and could be even negative for PN. This research was funded in part by the National Institute of Diabetes and Digestive and Kidney Diseases, grant DK093192 and NSF grant CBET‐0853157.
Homeostasis in undergraduate physiology textbooks
Authors: Michael Joel , McFarland Jenny , Cliff William , Modell Harold , Wenderoth Mary Pat , Wright Anne
Published: June 2021

Physiology faculty regard homeostasis as one of the core concepts their students should understand and be able to apply. We asked how 10 commonly used undergraduate physiology and biology textbooks introduce and use this core concept. We evaluated these textbooks to answer a number of questions: (1) do they contain an identifiable introductory ‘section’ on homeostasis, (2) is this initial description (terms and figures) complete and accurate, (3) to what extent are the same terms and similar figures used in different textbooks, and (4) do subsequent chapters apply the ideas about homeostasis. Two preliminary findings are of interest. First, the language used to talk about homeostasis is inconsistent, even in the same textbook, often confusing and sometimes incorrect. Second, even when a suitable description of homeostasis is present, the same language and diagram(s) may not be used to reinforce student learning in describing homeostasis in organ system physiology. The consequences of these problems for student understanding and application of homeostasis are discussed. Supported by NSF grant DUE‐1043443.
Point mutations in Escherichia coli DNA pol V that confer resistance to non‐cognate DNA damage
Authors: Hawver Lisa Ann , Tehrani Mohammad , Kania Danielle , Muser Stephanie , Beuning Penny J
Published: June 2021

When DNA is damaged, replicative DNA polymerases stall at the site of damage and DNA replication is compromised. One of the pathways in E. coli to cope with such damage is the SOS response that regulates the expression of at least 57 genes, including umuDC and dinB, which encode the Y family DNA polymerases UmuD′2C (pol V) and DinB (pol IV), respectively. UmuD′2C (pol V) is composed of the cleaved form of UmuD, UmuD′2, and the polymerase subunit UmuC. UmuD′2C's cognate lesions are abasic sites and thymine‐thymine dimers, while DinB specializes in bypassing N2‐dG adducts such as N2‐furfuryl‐dG. Y family DNA polymerases can also replicate undamaged DNA in an error‐prone manner and therefore play a role in mutagenesis and antibiotic resistance. Using hydroxylamine, we generated a library of UmuC mutations from which we then selected those that had the ability to confer survival to nitrofurazone, which is believed to cause predominantly N2‐furfuryl‐dG lesions. We tested in vivo and in vitro the ability of these selected UmuC variants to confer survival after UV irradiation as well their ability to bypass N2‐furfuryl‐dG, and discovered that the variants may not have changed specificity of Pol V, rather, they seem to disrupt protein‐protein interactions such as UmuC‐UmuD/D′ that are essential for Pol V function. Supported by Res. Corp. for Sci. Adv., Amer. Can. Soc., and NSF MCB‐0845033.
Peptide bond cleavage adjacent to aspargine or glutamine
Authors: Urbanski Laura M. , Chin Stacy L. , Reitter Julie N. , Mills Kenneth V.
Published: June 2021

Protein splicing is a post‐translational, self‐catalyzed process in which an intervening polypeptide, the intein, is self‐excised from the flanking polypeptides, or exteins. The third step of the splicing process is the cyclization of Asn or Gln coupled to peptide bond cleavage. We have studied this cyclization in model peptides with an N‐terminal Abz fluorophore and a C‐terminal Lys‐DNP quencher. Cyclization of Asn or Gln coupled to peptide bond cleavage results in fluorescence, which allows us to measure the rate of cyclization. The cyclization and cleavage can also be followed by HPLC and high resolution LC‐MS. Initial data suggest that the rate of Asn cyclization is significantly faster than Gln cyclization, that the rate of cyclization is increased by the influence of an upstream His, and that the cyclization is pH dependent.

This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and by the Camille and Henry Dreyfus Foundation.
Protein splicing of inteins from Synechococcus sp. PCC 7002 and Trichodesmium erythraeum
Authors: Siegart Nicolle M. , Urbanski Laura M. , Karanja Kevin , Colelli Kathryn M. , Reitter Julie N. , Mills Kenneth V.
Published: June 2021

Protein splicing is a four‐step intramolecular reaction, in which an intein flanked by N‐ and C‐terminal exteins is removed, resulting in ligation of the exteins. We are studying inteins that interrupt genes from the cyanobacteria Synechococcus sp. PCC 7002 (Ssp 7002) and Trichodesmium erythraeum (Tery). The Ssp 7002 intein has a highly conserved C‐terminal Asn, unlike an intein from Tery that has a C‐terminal Gln but also has 63% sequence identity with the Ssp 7002 intein. Both inteins splice with their native C‐terminal residue. Initial data suggest that Asn can substitute for Gln with the Tery intein, but Gln cannot substitute for Asn in the Ssp 7002 intein. Each intein also has conserved Cys residues that result in splicing controlled by disulfide bond formation in inteins from archaebacteria; we are currently determining if such redox‐state regulation is relevant with these cyanobacterial inteins as well.

This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and by the Camille and Henry Dreyfus Foundation.
Seeing Terrestrial Avian Locomotion with X‐ray Eyes
Authors: Kambic Robert , Gatesy Stephen
Published: June 2021

Birds, like humans, are obligate bipeds. Although most can fly, many birds are able walkers and runners. Avian terrestrial locomotion has implications for understanding structure and function in organisms, reconstructing how extinct dinosaurs lived, and pathological models. However, until recently, studying avian locomotion has been hampered by our inability to accurately measure joint and bone motion. Here we present 3‐D kinematics data from a chicken‐like bird, the helmeted guineafowl (Numida meleagris), using a new motion analysis method known as XROMM (X‐ray Reconstruction of Moving Morphology).

XROMM combines CT or laser‐scanned bone models with bi‐planar fluoroscopy to reconstruct six‐degree of freedom motion for skeletal elements. These highly accurate animations provide our first look at how birds move in 3‐D.

We find that long axis rotation of the femur and tibiotarsus is an important and underappreciated component of both steady and maneuvering locomotion. Understanding this rotation is critical to understanding how birds move through their environment, how hind limbs are designed, and how extinct theropods may have moved.

Grant Funding Source: NSF
Evolution of the in‐series circulation in tetrapods: form follows function
Authors: Owerkowicz Tomasz , Hicks James W.
Published: June 2021

The ancestral in‐parallel circulation of tetrapods combines a single ventricle and two systemic aortae, which allow central vascular shunting. Loss of the shunt and evolution of the in‐series circulatory design, as exemplified by extant mammals and birds, involved independent acquisition of two anatomic features: a complete interventricular septum, and a single aorta. Extent and location of the interventricular septum is dictated by the expression gradient of Tbx transcription factors in the myocardium. Loss of an aorta, however, has yet to be addressed by developmental studies. Our experimental work on the American alligator suggests that the heart's outflow tract is remodelled by changes in blood flow distribution in response to environmental stressors. Flow capacity of the left aorta (a conduit for the pulmonary bypass shunt) was significantly reduced (−27%) in alligator hatchlings chronically exposed for three months to atmospheric hypoxia (12%O2), when compared to conspecifics reared in normoxia (21%O2) or hyperoxia (30%O2). In contrast, outflow tract capacities did not differ in juvenile alligators subject to 15 months of different exercise regimina (sedentary, running or swimming). We suggest that a global hypoxic episode, rather than increased activity, may have been the driving pressure behind loss of the shunt and origin of the in‐series circulation. Funded by NSF IOB0445680 and IOS922756 to JWH.
Composition of Beverages in the Homes of Low‐Income Families in Greater Hartford, CT
Authors: Wakefield Dorothy B , Ferris Ann , Lora Karina
Published: June 2021

Baseline data from a longitudinal study describes an inventory of available beverages in the homes of 462 multicultural low‐income caregivers of 3–5 y. old children. Caregivers provided demographics and completed a home beverage inventory (HBI) at baseline, 3 and 6 months. HBI categories included 100% fruit juice (FJ), milk (flavored/unflavored/soy milk), juice drinks (juice drink, iced tea, energy drinks) (JD), regular and diet soda, bottled water (BW), low calorie 100% juices (LCFJ), and other (coffee/tea/supplements). The average HBI was comprised of 23% milk, 21% FJ, 18% JD, 17% BW, 8% regular soda, 3% LCFJ, 1% diet soda, and 9% other. The composition of the HBI was associated with caregiver's age. Older caregivers had more FJ, milk, and diet soda; and less JD and regular soda than younger caregivers (p<0.05). Families on SNAP had more milk (p=0.01) and less regular soda (p=0.01) than families not on SNAP. There were differences in JD (p=0.001), LCFJ (p=0.03), regular (p<0.001) and diet soda (p<0.001) by ethnicity (African American, Latino, White, West Indian, Other). Caregivers with <HS education had more regular soda than those with more education (p=0.01). Caregivers who were overweight/obese (p=0.01) and caregivers who were food secure (p=0.007) had less BW than underweight/normal or food insecure caregivers. Caregiver characteristics are associated with the composition of the beverages in the home.

Grant Funding Source: Funding: USDA/NRI 2008–55215‐19071
The Latin American Household Food Security Measurement Scale (ELCSA) was used to determine food security status in Honduran families with children ages 6 to 60 months
Authors: Chicoine Amanda Lynn , Kemmer Teresa M , Coello Miguel
Published: June 2021

The ELCSA survey was used to determine food security status in Santa Maria and Montaña de la Flor health regions. Households with children ages 6–60 months were randomly selected to participate. The level of food security was determined using the positive responses to the 15 question survey: no positive responses defined food secure, 1–5 defined food insecure, 6–10 defined very food insecure, and 11–15 defined severely food insecure. Within the 140 participating households, 11% of the households were food secure; 14% food insecure, 23% very food insecure, and 53% severely food insecure. Food insecurity was significantly (p ≤ 0.05) associated with having an underweight child (weight‐for‐age z‐score <−2) and telephone ownership; very food insecure with gender of child, living >; one hour from health clinic, electricity, and number of rooms in the house; and severe food insecurity with having an underweight child, child gender, telephone, number of rooms in the house, receiving food assistance, and electricity. Within the rural regions of Honduras evaluated, a very high prevalence of food insecurity exists and is associated with a variety of demographic and socioeconomic factors. These results will assist the Honduran Ministry of Health in directing resources to the heath regions most in need of assistance.

Grant Funding Source: USDA NIFA Competitive Grant 2011 67002 30202 and USDA, SD Agricultural Experiment Station Grant SD00H249–08
Zinc deficiency and growth status in children aged 6–60 months in rural Honduras
Authors: Lauseng Megan M. , Kemmer Teresa M. , Coello Miguel
Published: June 2021

Zinc is an essential trace mineral and its importance in growth and immune function has been established in young children; however, little is known about the prevalence of zinc deficiency in Honduras. The purpose was to determine the prevalence of zinc deficiency and its association with growth status within children ages 6–60 months in rural Honduras. The randomized cross‐sectional study was conducted in two rural Honduran health regions. Hair samples were used to determine zinc status. <−2 z‐score was used to determine stunting (height‐for‐age); wasting (weight‐for‐height); and underweight (weight‐for‐age). Zinc deficiency was defined as <70 μg/g and marginal deficiency as 70–100 μg/g. Bivariate correlations and chi square were used to determine zinc's association with growth status. 217 children aged 6–60 months (56.7% female) participated. Hair samples were obtained from 179 children. 15% of children were zinc deficient and 26% were marginally deficient. 60% of children were stunted, 4% wasted, and 16% underweight. No statistically significant associations were seen between zinc deficiency and growth status or gender. There was a high prevalence of zinc deficiency and a very high prevalence of stunting within children in the regions evaluated; therefore, additional research is required to determine prevalence of zinc deficiency and its associations with growth status throughout other regions of the country.

Grant Funding Source: USDA NIFA Competitive Grant 2011–67002‐30202 and USDA, SD Agricultural Experiment Station Grant SD00H249–08.
Elevated hair lead associated with anemia and socioeconomic status in Honduran children ages 6–60 months
Authors: Hemmingson Kaitlyn R , Kemmer Teresa M , Droke Elizabeth , Coello Miguel
Published: June 2021

A relationship exists between anemia and elevated lead levels; both are conditions that have proven to significantly impact health worldwide. A randomized cross‐sectional study of 217 children ages 6–60 months was conducted in Santa Maria and Moñtana de la Flor, Honduras. The purpose was to determine the relationship between hair lead and anemia in children, and to determine if socioeconomic factors are associated with high lead levels. 20.6% of the population had elevated (≥ 5–10 μg/dL) hair lead and an additional 5.6% showed significant elevation (≥ 10 μg/dL). 33.5% were anemic (Hb < 11g/dL). A significant association was found between elevated hair lead and anemia (p=0.01). 60.5% of the participants were stunted, 30.7% were underweight, and 4.7% were wasted. Socioeconomic indicators of high lead levels included: living in Santa Maria, electricity, telephone ownership and type of stove. Additional research is required to determine the prevalence of elevated lead status throughout Honduras.

Grant Funding Source: USDA NIFA Competitive Grant 2011興67002興30202 and SD Agricultural Experiment Station HATCH Grant SD00H249–08
The Breastfeeding and Employment (BESt) Study: New mother employee perceptions of breastfeeding support in companies
Authors: Olson Beth , Braun Michael
Published: June 2021

Working women breastfeed at lower rates than the unemployed, but little is known on their perceptions of breastfeeding support in companies.

Objective

Determine the role of company policies and manager attitudes on new mother employee perceptions of workplace breastfeeding support.

Methods

The BESt Study surveyed companies via mailings to Human Resource Managers with surveys on company's breastfeeding supports and for matched employee and manager pairs. Companies received a total support score and subscores. Analysis was by hierarchical linear modeling.

Results

Employee scores were not associated with any company scores. Employees with greater job autonomy, a better physical environment and female managers were more positive (p<0.01). Total company scores were associated with more positive manager attitudes, due to structural supports (P<0.01) but not employee supportive supports. In a combined model, positive manager attitudes was associated with more negative perceptions of employees (P<0.05). This relationship was marginally significantly moderated by employees’ prior breastfeeding experience and moderated by managers’ previous experience with breastfeeding employees (P<0.05).

Conclusions

New mother employees may be more influenced by their ability to control their work environment, and benefit from their own breastfeeding experience as well as that of their manager.

Funded by USDA NRI 2006–35215‐16703