bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of estrogen on statin‐induced toxicity in proliferating and differentiating myoblasts
Authors: Mertz Chelsea Claire , Bielman Marie , Ball Karen L
Published: June 2021

Estrogen (E) regulates apoptosis in a variety of cell types with antiapoptotic effects reported in skeletal myoblasts. Interestingly, it has been suggested that in vivo females are more susceptible to myotoxicity from statin use. Our aim was to assess the role of E in simvastatin (SS) induced apoptosis in myoblasts. C2C12 cells were cultured in growth media (GM) and plated in 96‐well plates. After 72 hrs exposure to GM, GM + SS (10 μM), or GM + SS + E (1, 0.1, 0.01, 0.001, 0.0001 μM) an MTS cell viability assay was run. In a separate experiment cells were stained with DAPI to quantify apoptotic nuclei. To measure SS + E effects in differentiating myoblasts, cells were maintained in GM until confluent then exposed to the following differentiation media: DM, DM+ SS (10 μM), or DM+ SS + E (0.1, 0.01, 0.001 μM) for 72 hours. Cells were DAPI‐stained and apoptotic nuclei quantified. Data was analyzed by 1‐way ANOVA; p<0.05. As in previous studies, SS decreased the number of viable cells to 45% of control. However, E had no effect at any concentration. DAPI staining showed induction of apoptosis in the presence of SS with E neither blocking or enhancing the effect. In differentiating cells, apoptotic nuclei increased from 6.5% to over 50% with SS. Again, E had no effect. Further studies are needed to explain this lack of E effect in vitro with reported in vivo female susceptibility to statin‐induced myotoxicity.

Supported by NSF STEP Grant
Activation of both phasic and tonic GABAA¬ receptors modulates vagal efferent outflow regulating gastric motor functions
Authors: Reagan Zachary K , Browning Kirsteen N
Published: June 2021

The dorsal motor nucleus of the vagus (DMV) provides the parasympathetic innervation to the stomach. DMV neurons display both synaptic and extrasynaptic GABAA receptors that are activated by phasic and tonic GABA release, respectively. Studies have demonstrated that ongoing activation of GABAA receptors is critical in determining vagal efferent outflow to the stomach. The present study was designed to investigate whether phasic and/or tonic GABAA receptors are involved in the regulation of gastric tone.

Miniaturized strain gauges were sutured to the ventral surface of the gastric corpus in male Sprague‐Dawley rats (175–300g). The GABAA receptor antagonists, gabazine (0.78–6.25pmol) and/or bicuculline methiodide (BMI; 25–100pmol), were microinjected into the dorsal vagal complex and the increase in gastric tone expressed relative to the maximal increase induced following i.v. application of the non‐selective muscarinic agonist, bethanecol (352pmol; 1ml). At low doses, gabazine acts selectively on extrasynaptic receptors, whereas BMI acts on both synaptic and extrasynaptic receptors.

Gabazine increased gastric tone 5.1±1.7% at 0.78pmol, n=10, increasing to 15.3±1.74% and 19.1±5.94% in response to 3.125 and 6.25pmol, n = 5,6, respectively (P<0.05 vs 0.78pmol for each). At lower doses, BMI induced similar increases in gastric tone (11.0±4.7% and 14.9±5.5% at 25 and 50pmol, n = 8,8, respectively) whereas the response was significantly greater at 100pmol (35.7±17.8%, n=4, P<0.05). These data suggest that the activity of DMV neurons, hence vagal efferent outflow to the stomach, is regulated by both synaptic and extrasynaptic GABAA receptors implying a more complex synaptic modulation than previously considered.

Supported by NSF IOS 11489364
α1A adrenergic receptor influences on progenitor cell fate in the adult hippocampus
Authors: Collette Katie , Nielsen Amber , Perez Dianne , Doze Van
Published: June 2021

Long‐term stimulation of α1A adrenergic receptors (α1AARs) increases the number of proliferating cells in the subgranular zone (SGZ) of the adult dentate gyrus and enhances learning and memory in the mouse. The creation of new neurons in the SGZ of the adult brain has been linked with improved learning and memory in mouse, rat, and primate models. The role of the new neurons in learning and memory has not been fully examined though some studies show that the new cells play different roles as immature and mature cells. We used transgenic mouse models to determine the effect of chronic α1AAR activation on differentiation and survival of the adult‐generated cells in the hippocampus. Animals were injected with BrdU to label dividing cells and perfused 4 weeks later to allow time for the new cells to differentiate. Immunohistochemical markers were used to identify the new cells which included excitatory granule cells, astrocytes, and a very small number of the interneurons. Supported by North Dakota EPSCoR IIP Seed Award Grant EPS‐081442, National Center for Research Resources Grant P20RR016741 and National Institute of General Medical Sciences Grant P20GM103442 from the NIH, NHLBI Grant RO1HL61438, NSF Graduate Research Fellowship Program, and NSF MRI Grant 0619688.
Source identification of QT interval changes in diabetic patients
Authors: Wang Siqi , Evans Joyce M , Knapp Charles F , Schlegel Todd T , Elayi Samy C , Patwardhan Abhijit R
Published: June 2021

We acquired experimental 12‐lead ECG and developed a hearttorso computer model to simulate 12‐lead ECG to 1) reproduce QT interval prolongation seen in diabetic patients, and 2) describe a possible mechanism of QT interval prolongation by linking QT interval to regional cardiac sympathetic denervation at the cellular level in the model. Twelve‐lead ECG were acquired in 11 able‐bodied subjects (AB), 7 diabetics without (D0), 7 diabetics with mild (D1) and 8 diabetics with severe (D2) neuropathy. Heart rate‐corrected QT interval (QTc) was computed. Action potentials were modeled phenomena logically. Twelve‐lead ECG were produced as forward solutions from equivalent cardiac source. Different patterns of regional denervation taken from previous studies of cardiac images of diabetic patients, guided the simulation of pathological changes, generating 12‐lead ECG from every trial. The minimum QTc interval from the lateral leads was longer in D2 than in AB. Prolonging action potential duration (APD) in the basal septal region in the model produced ECG and QT interval similar to that of D2 subjects. We conclude that minimum QTc interval of lateral leads was more diagnostic than that from other leads. The simulation suggested that QT interval prolongation in our diabetic patients with neuropathy may have been due to APD prolongation in basal septal region, secondary to sympathetic denervation in this region. This study was supported by NIH RO1 NS39774, Kentucky NASA EPSCOR WKU 52611, UK GCRC USPHS grant M01RR02602, and the University of Kentucky Center for Clinical and Translational Sciences.
Preservation of skeletal muscle by the heat shock proteins following frostbite injury
Authors: Mestril Ruben , Schweigert Colin , Batey Jason , Liskutin Tomas , Baldwin John
Published: June 2021

Background

Skeletal muscle damage due to frostbite represents a serious medical problem. Oxidative stress and inflammatory response cause muscle degeneration following skeletal muscle injury. Studies have shown that the heat shock proteins (hsp) are able to protect against oxidative stress and inflammation.

Results

In the present study we used rat hindlimbs to examine the effects of several heat shock protein inducers, such as radicicol, 17‐AAG and 17‐DMAG following frostbite injury. Our results show that rat hindlimb muscles injected with these hsp inducers following frostbite injury exhibit less damage, less hypercontracted muscle fibers and less inflammatory cell infiltration as compared to control rat hindlimb muscles. We also find that these heat shock protein inducing compounds are able to conserve muscle function even when administered after frostbite injury has occurred. These results suggest that hsp inducing compounds are able to protect skeletal muscle if administered in the early stages following frostbite injury.

Conclusion

We conclude that compounds that induce heat shock proteins are able to preserve skeletal muscle function and structure if injected shortly after frostbite injury. Our studies provide the basis for the development of therapeutic strategies to treat injury caused by frostbite involving severe muscle damage.

This research was supported by an award from the USAMRMC.
The energy content of restaurant foods without stated calorie information
Authors: Urban Lorien E. , Lichtenstein Alice H. , Gary Christine E. , Fierstein Jamie L. , Equi Ashley , Kussmaul Carolyn , Dallal Gerard E. , Roberts Susan B.
Published: June 2021

National recommendations emphasize self‐monitoring for prevention and treatment of obesity; however, little information is available on the dietary energy contents of meals obtained from non‐chain restaurants, which account for ≈50% of restaurants in the US. Using bomb calorimetry, we determined gross energy of the 42 most popular meals from 34 randomly selected restaurants in the 9 most common ethnic restaurant categories in Massachusetts. Mean energy of the 158 meals was 1327 kcal (95% CI, 1248–1406 kcal), equivalent to 66% of typical daily energy requirements. All individual meal categories provided substantially more energy than required for weight maintenance, 95% of meals provided >;25% of typical daily energy requirements, and 7% provided >;100%. There was a significant effect of ethnic category on meal energy (P≤.(standardized to gross energy) than equivalent non‐chain entrees in the current study (P=.18), and meals contained 16% more energy than national food database information for equivalent meals (P=.0002). Non‐chain restaurants may be important contributors to the obesity epidemic because they provide large amounts of dietary energy that are incompatible with weight management, and at the same time do not typically report any nutrition information. Funding: USDA agreements 58–1950‐0–0014, 1950–51000‐072–02S.

Grant Funding Source: USDA agreements with Tufts University
Clock 3111 T/C SNP Interacts with Saturated Fatty Acid Intake to Modulate Plasma LDL‐C Concentrations in the Boston‐Puerto Rican Health Study
Authors: Dashti Hassan S , Tucker Katherine L. , Smith Caren E. , Ma Yiyi , Lee Yu‐Chi , Ordovás José M.
Published: June 2021

Disruption of circadian rhythms is associated with metabolic syndrome (MetS) traits and dyslipidemia and could be induced by variants of circadian genes and a high‐fat diet. While the Clock 3111 T/C single nucleotide polymorphism (SNP) has been reported to interact with saturated fatty acid (SFA) intake to modulate MetS traits, no study has reported the interaction of this SNP with fat intake on dyslipidemia outcomes, such as elevated LDL‐C concentrations. The aim of the current study was to examine whether the association between Clock 3111 T/C SNP and plasma LDL‐C concentrations is modulated by SFA intake. Participants in the Boston‐Puerto Rican Health Study (n=772) were examined for interaction using multivariate linear regression models, where a dominant genetic model was applied. While LDL‐C did not differ between TT participants and carriers of the minor C allele (TC and CC), by dichotomizing SFA intake, we identified a significant gene‐diet interaction associated with plasma LDL‐C concentrations at this locus (P = 0.01). In the high SFA intake group (≥ 21g/d), the minor C allele was protective against elevated plasma LDL‐C concentrations (P = 0.02). This gene‐diet interaction provides additional evidence for the role of a high‐fat diet in regulating chronobiological functions and, if replicated in additional populations, may be of application for future personalized nutritional recommendations.

Grant Funding Source: U.S. Department of Agriculture
Placental expression of the heme scavenger receptor, LDL receptor‐related protein 1, is associated with expression of placental heme exporter, feline leukemia virus C receptor 1
Authors: Cao Chang , Cooper Elizabeth , Pressman Eva , Guillet Ronnie , O'Brien Kimberly O
Published: June 2021

LDL receptor‐related protein 1 (LRP 1) is the surface receptor for the plasma heme‐hemopexin complex and is involved in heme clearance following intravascular red cell destruction. Both LRP1 and the heme exporter feline leukemia virus C receptor 1 (FLVCR1) are highly expressed in the human placenta and may play a role in partitioning heme iron between the maternal and fetal compartments. To explore this possibility, protein expression of LRP1 (n=122) and FLVCR1 (n=62) were assessed in pregnant adolescents (13–18 y) in relation to plasma concentrations of the heme scavenger, hemopexin, and iron status indicators (hemoglobin, serum ferritin, transferrin receptor, and hepcidin) at mid‐gestation (~26 wks) and in maternal/cord blood at delivery. Placental LRP1 expression was positively associated with placental FLVCR1 (r=0.41, n=62, p<0.01), maternal serum transferrin receptor at delivery (r=0.20, n=117, p=0.03), and cord hepcidin (r=0.22, n=113, p=0.02). Greater decreases in hemopexin across pregnancy were associated with higher placental LRP1 (r=0.33, n=55, p=0.01). Placental LRP1 is associated with placental FLVCR1 and a maternal index of intravascular hemolysis. Further studies are needed to elucidate functional roles of LRP1 in placental iron transport.

Supported by: USDA 2008–0857

Grant Funding Source: USDA
Usability of mobile phone food records to assess dietary intake in adolescents
Authors: Casperson Shanon L. , Reineke Jessica , Sieling Jared , Moon Jon , Roemmich James
Published: June 2021

Mobile technologies are emerging as a valuable tool to collect and assess dietary intake. Adolescents readily accept and adopt new technologies, hence, a food record application (FRapp) may provide an accurate mechanism to monitor dietary intake. We examined the usability of a FRapp in 17 free‐living adolescents. Participants (11 – 14 y) received detailed instructions and were asked to capture before and after images of all foods and beverages consumed, and enter text descriptions using the FRapp for 3–7 days. Sixteen participants used the FRapp and 1 used the phone's native camera function to capture food images. A total of 109 meals were recorded, of which 40% included a post meal image. 62% of images included all foods, beverages and a fiducial marker. Text was recorded for 10 meals without images. Only 6 participants recorded 3 meals a day. One participant reported the FRapp as being too difficult (1 meal recorded) and another recorded all meals as snacks. Of the meals recorded, 25, 24, 25, and 35 were breakfast, lunch, dinner, and snacks, respectively. These data suggest that a FRapp can be useful for dietary intake assessment in adolescents. A detailed interactive training session, including a test meal, may increase the inclusion of all foods and beverages in the images. Additionally, a trigger is needed to remind participants to take a post image.

Grant Funding Source: ARS‐USDA
Drill Sergeants’ two contrasting approaches to “soldierization” and instilling healthy eating behaviors in soldiers
Authors: Blake Christine E , Rosemond Tiara N , Torres‐McGehee Toni M , Frongillo Edward A , Burke Michael P , Lenkerd Allison , Minton Dawn M , Cable LTC Sonya , Glover Saundra H
Published: June 2021

Adult leaders such as athletic coaches, teachers, and drill sergeants (DS) play a key role in instilling healthy habits as adolescent's transition to adulthood. Positive approaches have been promoted in coaches and teachers, but little is known about DS. This study aimed to understand DS perspectives on instilling healthy eating behaviors in soldiers as part of their overall approach to basic training. In‐depth qualitative interviews conducted with 30 DS at two US Army training bases were transcribed verbatim and analyzed using a grounded theory approach. DS varied in their perceptions of the importance of nutrition, responsibility for promoting nutrition, general approach to training, and integration of nutrition into training. Two general approaches to what DS called “soldierization” emerged. One included mentoring, role modeling, and tailoring strategies to the individual. The other was less responsive to individual differences, and emphasized obedience, discipline, and intimidation. General approaches framed strategies for instilling healthy eating behaviors in soldiers. Many DS currently use such strategies in soldiers that are compatible with best practices used by successful athletic coaches and teachers, but further improvements would potentiate the positive impact that DS could have on soldiers’ health eating behaviors. Funded by Department of Defense.
Lycopene metabolite, apo‐10′‐lycopenoic acid, inhibits diet‐induced obesity‐promoted liver carcinogenesis via the up‐regulation of SIRT1 signaling and the decrease in liver inflammation.
Authors: IP Blanche C , Hu Kang‐Quan , Liu Chun , Smith Donald E. , Ausman Lynne M. , Wang Xiang‐Dong
Published: June 2021

Chronic inflammation caused by diet induced‐obesity (DIO) is associated with increased liver cancer risk. We previously demonstrated that dietary lycopene ameliorated DIO‐promoted liver precancerous lesions, while supplementation of lycopene's cleavage metabolite, apo‐10′‐lycopenoic acid (APOLA), upregulated hepatic sirtuin 1 (SIRT1) activity. SIRT1 is a NAD‐dependent deacetylase that inhibits DIO‐promoted liver inflammation and carcinogenesis when overexpressed. This study aimed to elucidate the chemopreventative effects of APOLA and associated SIRT1 signaling with in‐vitro and in‐vivo models. APOLA treatment in immortalized liver cells upregulated SIRT1 expression and AMP‐activated protein kinase‐á (AMPKá) phosphorylation in a dose‐dependent manner. Dietary APOLA at 10 mg/kg of diet for 24 weeks significantly reduced carcinogen‐initiated, DIO‐promoted liver cancer (in tumor multiplicity and volume) and lung tumor incidence in C57BL/6J mice. These outcomes were associated with reduced liver inflammation (decreased cleavage of caspase‐1, IL‐6, TNFá, NF‐êB p65 protein expression, and inflammatory foci), and increased hepatic AMPKá phosphorylation, SIRT1 protein expression and its deacetylase activity. Taken together, this present study revealed that APOLA inhibits DIO‐promoted liver carcinogenesis by stimulating AMPK‐SIRT1 signaling and reducing liver inflammation.

Grant Funding Source: USDA/ARS Grant 1950–51000‐064S
Differential response of anabolic signaling with high‐fat feeding and aging in skeletal muscle after insulin stimulation
Authors: McDonald Devin J , Morris Evan P , Haran Prashanth H , Fielding Roger A , Rivas Donato A
Published: June 2021

The increased accumulation of intramyocellular lipids with advancing age and obesity may be a contributing factor to anabolic resistance in skeletal muscle. Aging and obesity have differential causes of fat accumulation in muscle and it is hypothesized they may have divergent mechanisms leading to anabolic resistance. In order to test this hypothesis we used C57BL/6 mice aged 6 months and fed a control diet (YNG CFD, 15% fat/kcal) or a diet high in fat (YNG HFD, 60% fat/kcal) and mice aged 24 months fed a control diet (OLD CFD) for 12 weeks. The animals were then fasted 5 h and were either injected with a single bolus of insulin (INS, 0.85 IU/kg bw) or saline vehicle (SAL) 30 mins before skeletal muscle collection. Western blotting was carried out to detect differences in activation of growth and degradation pathways. The phosphorylation Akt, mTOR and S6K1 were higher with INS in all groups but the magnitude of this change was differentially affected by age and diet. INS did not change the phosphorylation of FOXO1/FOXO4, NFβB and its inhibitor IκBα. However, IκBα phosphorylation, and therefore inactivation, was higher in OLD CFD in SAL and INS groups. Using quantitative PCR we observed a significant difference in the gene expression of NFβB targets, TLR2 and IL1β with aging and diet. We conclude that aging and a high‐fat diet differentially effect growth and degradation pathways in skeletal muscle after insulin stimulation.

Grant Funding Source: USDA and NIH/NIA
The association between diet quality, BMI and health‐related quality of life in the Geisinger Rural Aging Study (GRAS)
Authors: Ford Dara J , Jensen Gordon L , Bailey Regan , Smiciklas‐Wright Helen , Erickson Pennifer , Wood Craig , Still Christopher , Coffman Donna , Hartman Terryl J
Published: June 2021

Nutrition status is a predictor of quality of life. This study sought to determine the association between diet quality, BMI, and health‐related quality of life (HRQOL) in a population of adults ≥74 y. The Diet Screening Tool (DST) is a 25 item questionnaire scored from 0 (poor) to 100 developed to assess diet quality in older adults. The Health and Activity Limitations Index (HALex) is a HRQOL measure scored on a 0.0 (death) to 1.0 (optimal health) scale. A total of 5,993 GRAS participants were mailed HRQOL and DST questionnaires with 2,975 (1260 male, 1715 female; mean age 81.4 ± 4.4) providing complete data. Multivariate linear regression models were used to analyze associations between HALex score, BMI and DST score after controlling for gender, age, education, and smoking. The mean adjusted HALex score for participants in the lowest quintile of DST scores was 0.72 ± 0.2 compared to 0.77 ± 0.2 for those in the highest quintile of DST Scores (p‐trend <0.01). Adjusted mean HALex scores were significantly lower for obese (BMI ≥30; 0.72 ± 0.21, P=0.006) and underweight individuals (BMI <18.5; 0.64 ± 0.23, P=0.02) and significantly higher for overweight individuals (BMI 25–29.9; 0.76 ± 0.18, P=0.02) compared to those with BMI 18.5–24.9 (0.73 ± 0.22). Poor diet quality, as assessed by the DST, is associated with lower HRQOL in older adults. USDA #1950–51530‐010–00

Grant Funding Source: USDA
Calcium homeostasis and bone metabolic responses to protein diets and energy restriction: a randomized control trial
Authors: Cao Jay , Pasiakos Stefan M , Margolis Lee M. , Whigham Leah D. , McClung James P. , Young Andrew J. , Combs Gerald F.
Published: June 2021

Despite some beneficial effects on bone, high protein diets have been conventionally considered a primary dietary risk factor for osteoporosis due to the acid load associated with protein catabolism. To test the hypothesis that high dietary protein diets do not negatively affect calcium homeostasis and bone turnover, 32 men and 7 women consumed diets providing protein at 0.8 (RDA), 1.6 (2X‐RDA), or 2.4 (3X‐RDA) g·kg‐1·d‐1 for 31 days in a randomized‐block design. Ten‐day of weight maintenance (WM, days 1–10) preceded 21‐day of energy deficit (ED, days 11–31), during which energy intake was restricted to 60% of WM. Ca absorption (ratio of 44Ca:42Ca) and circulating indices of bone turnover and were determined during WM (day 8) and ED (day 29). Regardless of energy state, urinary pH was lower (P < 0.05) for 2X‐RDA (6.28 ± 0.05) and 3X‐RDA (6.23 ± 0.06) compared to RDA (6.54 ± 0.06). ED induced weight loss (3.2 ± 1.0 kg body mass, P < 0.05), and a 13% reduction in serum IGF‐I (P < 0.01). Remaining markers of bone turnover, as well as urinary Ca excretion, absorption, bone mineral density and content were not affected by protein level and ED (P >; 0.05). These data demonstrate that consuming high protein diets do not disrupt Ca homeostasis and skeletal integrity, whereas reductions in IGF‐1 may suggest diminished bone formation during sustained ED.

The study was supported by USDA ARS and USAMRMC.
Ghanaian caregivers’ opinions on feeding 2‐ to 5‐year old children varies by agro‐ecological zone
Authors: Christian Aaron Kobina , Colecraft Esi , Marquis Grace S , Lartey Anna , Sakyi‐Dawson Owuraku , Butler Lorna M , Ahunu Ben , Reddy Manju Bokka , Jensen Helen H , Huff‐Lonergan Elisabeth J
Published: June 2021

Childhood malnutrition is high in Sub‐Saharan Africa. Knowing caregivers’ opinions about feeding children may aid in designing effective nutrition interventions. Data on caregiver feeding opinions, and diets of 2‐ to 5‐y‐old children were collected from 608 caregivers living in three ecological zones (EZ) of Ghana. This analysis compared caregivers’ feeding opinions across three ecological zones. About 27.8% of caregivers believed that their children needed to be fed only 2 to 3 times/d. Reasons for adult supervision during child meal times, feeding diverse foods, and prioritizing a child to receive animal source foods (ASF), and the perceived child benefits of ASF differed across EZ (P<0.001). Caregivers’ opinions in agreement with recommended child feeding practices were scored as good opinion otherwise their opinion scored as poor. Children of caregivers with good feeding opinion scores consumed more diverse ASF compared to those of caregivers with poor opinions (5.0 ± 2.1 vs. 4.6 ± 2.1; P=0.026). Age and education of caregivers positively predicted better feeding opinions (P<0.01); living in the Guinea Savannah ecological zone was associated with poor feeding opinions. A key component to improving child nutrition is to understand the opinions held by caregivers in order to address them adequately within the specific locale of the caregiver and child. Support: GL‐CRSP, funded in part by US‐AID, PCE‐G‐00–98
Dietary wolfberry supplementation enhances protective effect of flu vaccine against influenza infection in aged mice
Authors: Du Xiaogang , Wang Junpeng , Niu Xinli , Smith Donald , Vidal Karine , Wu Dayong , Meydani Simin Nikbin
Published: June 2021

Current vaccines for influenza do not fully protect the aged against flu. Wolfberry, or goji berry, has been shown to improve immune response in the aged; however, it is not known if it would improve vaccine efficacy against influenza infection. Further, the underlying mechanisms are not known. Old mice were fed a diet containing 5% Lacto‐Wolfberry (LWB) for 30 d, immunized with the flu vaccine on days 31 and 52, and challenged with the influenza A/Puerto Rico/8/34. Results showed that LWB supplementation increased flu antibody titers and decreased influenza‐induced weight loss beyond that observed by vaccine alone (p<0.0001 for overall diet × vaccine interaction by ANOVA). Further in vitro mechanistic studies showed that wolfberry significantly enhanced maturation and activity of antigen presenting dendritic cells (DC) in aged mice, as measured by expression of cell surface markers CD40, CD80, CD86, and MHC‐II, their production of cytokine IL‐12 and TNF‐α, and endocytosis. Further, adoptive transfer of wolfberry extract‐treated bone marrow DC (loaded with OVA323–339‐peptide) from the aged mice promoted splenocyte proliferation, and increased antigen‐specific IL‐4 and IFN‐γ production in CD4+ T cells. In summary, our data indicate that LWB supplementation significantly enhances flu vaccine efficacy, at least in part, through improving DC function in aged mice. Supported by the USDA contract #58–1950‐0–014.
Very low food security in young citizen children correlates with mothers’ foreign birth; associations vary by mothers’ length of stay in the US
Authors: Cook John T , Frank Deborah A , Coleman Sharon , de Cuba Stephanie Ettinger , Pasquariello Justin
Published: June 2021

This study's objective was to test whether very low food security (VLFS) in children, the most severe food insecurity measured in the US, is related to mothers’ foreign birth, & whether mothers’ length of stay (LOS) in the US affects that relationship. Data are from ongoing Children's HealthWatch research on 40,460 children ages <48 months whose mothers were interviewed at pediatric clinics & hospital emergency departments in Baltimore, Boston, Little Rock, Los Angeles, Minneapolis, Philadelphia & Washington, DC June 1998‐December 2011. IRB approval was obtained at each site, & renewed annually. In multivariate logistic regression models adjusted for site & mothers’ characteristics, children of foreign‐born mothers had adjusted odds of having VLFS in children 194% greater than children of US‐born mothers. In models using data stratified by mothers’ LOS in the US, children of mothers in the US >;10 yrs had adjusted odds of VLFS in children 44% lower than children of mothers in the US <5 yrs. The goal of ending child hunger (VLFS in children) in the US by 2015 is unlikely to be achieved without targeted support for families with recent immigrant mothers. This project was supported with a grant from the University of Kentucky Center for Poverty Research through funding by the U.S. Department of Agriculture, Food & Nutrition Service, contract number AG‐3198‐B‐10–0028.
Enrichment of meiotic recombination hotspot sequences by avidin capture technology
Authors: Teixeira Daniel Camara , Zempleni Janos , Malkaram Sridhar A
Published: June 2021

About 40% of the hotspots for meiotic recombination contain the degenerate consensus sequence 5′‐CCNCCNTNNCCNC‐3′. Here we present a novel protocol for enriching hotspot sequences from digested genomic DNA by using biotinylated oligonucleotides and streptavidin‐coated magnetic beads. The captured hotspots can be released by simple digestion with restriction enzymes for subsequent characterization by second generation sequencing or PCR. The capture protocol specifically enriches hotspot sequences, judged by using fluorophore‐conjugated synthetic oligonucleotides and synthetic double‐stranded oligonucleotides in combination with PCR. The capture protocol enriches single stranded DNA, denatured double‐stranded DNA, and large fragments (>;3,000 bp) of digested plasmid DNA with good efficacy. No false positive and false negatives were detected when enriching digested DNA from human cell cultures and primary human cells. The protocol can be adopted to enriching sequences other than the hotspot sequence by altering the sequence in the capture oligonucleotide. We intend to apply this protocol in studies assessing effects of micronutrient status on meiotic recombination events in human sperm.

Grant Funding Source: ARD Hatch, NIFA and NIH.
Neuregulin‐1 overexpression and p53 haploinsufficiency cooperatively promote de novo MPNST pathogenesis
Authors: Brosius Stephanie Nicole , Turk Amy Nicole , Byer Stephanie Joy , Brossier Nicole Marie , Kohli Latika , Roth Kevin Aaron , Carroll Steven Lewis
Published: June 2021

Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive sarcomas arising from neurofibromas in patients with neurofibromatosis type 1 (NF1). We have shown that transgenic mice expressing neuregulin‐1 (NRG1) in Schwann cells (P0‐ GGFβ3 mice) develop neurofibromas and MPNSTs on a C57BL/6 × SJL background. However, on a C57BL/6J background, P0GGFβ3 mice fail to develop tumors despite persistent transgene expression and Schwann cell hyperplasia. Hypothesizing that tumor suppressor gene loss would complement NRG1 overexpression and rescue tumorigenesis, we created cohorts of P0GGFβ3 >;Nf1+/−, P0GGFβ3; Trp53+/− and control (P0‐GGFβ3, Nf1+/−, Trp53+/−) mice and compared their survival and tumorigenesis. The survival of P0GGFβ3; Nf1+/− mice was equivalent to controls and they did not develop tumors. In contrast, P0GGFβ3; Trp53+/− mice died earlier (mean survival, 232 days) and MPNSTs were present in 88.4% of these mice. Further, unlike outbred P0GGFβ3 mice, MPNSTs in P0GGFβ3; Trp53+/− mice arose de novo rather than from neurofibromas. Real‐time PCR, immunohistochemical and sequence analyses showed persistent wild‐type p53 expression. These findings show that NRG1 acts in the same pathways affected by Nf1 loss and suggest that Trp53 haploinsufficiency in the face of NRG1‐driven Schwann cell hyperplasia destabilizes the genome and promotes de novo MPNST tumorigenesis. Funded by NIH and DOD.
Climatic variation in human choanal shape: A 3D geometric morphometric study
Authors: Pagano Anthony Santino , Yuan Derek , Fischer Daniel , Laitman Jeffrey T.
Published: June 2021

There has been considerable study on climatic diversity in human piriform aperture dimensions but relatively little on the posterior nares or choanae. This study quantified shape differences in choanal morphology between two broad groups of “cold” and “tropical” climate human populations. The former consisted of Alaskan Inuits, Aleutian Islanders, Northern Chinese, and Northern Europeans (n=131) while the latter group was comprised of East and West Africans and South East Asians (n=63). Such broad groups were employed to control for any feature not directly related to climate (e.g. facial prognathism). Shape differences were quantified from landmark coordinate data collected via a Microscribe digitizer. After Procrustean superimpositioning, relative distances were compared via Student's t‐tests. The results showed that found that cold climate populations may be distinguished by significantly (p<0.0001) smaller choanal plane angles (more vertically oriented choanae), smaller maximum relative nasopharyngeal depth (staphylion‐sphenobasion chord), and higher relative medial pterygoid plate height. However, none of these features distinguished any single cold climate population from all other tropical populations. Climate likely influences choanal and overall nasopharyngeal morphology in a global context that is discernable only in large‐scale comparison of pooled populations.

Grant Funding Source: National Science Foundation
Overexpression of the lysophosphatidic acid generating enzyme autotaxin/ENPP2 potentially promotes the pathogenesis of neurofibromatosis type 1‐associated neurofibromas and malignant peripheral nerve sheath tumors
Authors: Peavler Lafe Timothy , Byer Stephanie J. , Roth Kevin A. , Carroll Steven L.
Published: June 2021

Aggressive Schwann cell‐derived sarcomas known as malignant peripheral nerve sheath tumors (MPNSTs) are the leading cause of death in patients with neurofibromatosis type 1 (NF1). It has been proposed that growth factors such as lysophosphatidic acid (LPA) interact with NF1 loss to drive MPNST pathogenesis. LPA promotes Schwann cell proliferation, migration and survival; LPA induced proliferation and survival is further enhanced in Nf1−/− Schwann cells. As autotaxin (ATX)/ENPP2, the enzyme that generates LPA via conversion of lysophosphatidylcholine, is overexpressed in several other tumor types, we hypothesized that ATX overexpression promotes the pathogenesis of MPNSTs and the benign neurofibromas from which they are derived. To test this hypothesis, we examined ATX expression in human neurofibromas, MPNSTs and MPNST cell lines and MPNSTs arising in a novel mouse model (P0‐GGFβ3 mice). Immunohistochemical studies showed that ATX was overexpressed in human and murine tumors compared to normal nerve. Real time PCR and immunoblots further demonstrated increased ATX expression in a panel of MPNST cell lines; the ATX transcripts present in these cells encoded several functionally distinct splice variants of ATX including α, β and γ isoforms. We are currently knocking down ATX expression and generating ATX overexpressing transgenic mice to validate ATX as a therapeutic target in MPNSTs. Funded by NIH and DOD.
The N‐terminal GIY‐YIG endonuclease activity of Arabidopsis glutaredoxin AtGRXS16
Authors: Cheng Ninghui , Liu Xi , Liu Shian , Feng Yingang , Deng Haiteng , Hirschi Kendal D. , Wang Xinquan
Published: June 2021

A chloroplastic monothiol glutaredoxin, AtGRXS16, comprises two distinct functional domains, an N‐terminal GIY‐YIG endonuclease motif (NTD) and a C‐terminal Grx module, to coordinate redox regulation and DNA cleavage in chloroplasts. Structural determination of AtGRXS16‐NTD showed that it possesses a GIY‐YIG endonuclease fold, but the critical residues for the nuclease activity are different from typical GIY‐YIG endonucleases. AtGRXS16‐NTD was able to cleave λDNA and chloroplast genomic DNA and the nuclease activity was significantly reduced in AtGRXS16. AtGRXS16‐NTD could inhibit the ability of AtGRXS16 to suppress the sensitivity of yeast grx5 cells to oxidative stress; however, AtGRXS16 with a Cys123Ser mutation were active in these cells and functional in Fe‐S cluster biogenesis. Furthermore, the two functional domains were shown to be negatively regulated through a formation of an intramolecular disulfide bond. These findings unravel a novel manner of regulation for Grxs and provide insights into the mechanistic link between redox regulation and DNA metabolism in chloroplasts. This study is supported by USDA Cris fund.
Crystallographic studies of biotin carboxylase bound to associated ligands
Authors: Broussard Tyler C , Kobe Matthew J , Pakhomova Svetlana , Neau David B , Price Amanda E , Champion Tyler S , Waldrop Grover L
Published: June 2021

Acetyl‐CoA carboxylase (ACC) is responsible for the first committed step in fatty acid synthesis. Therefore, ACC is a target for novel antibiotic development. Previous attempts to develop an antibiotic have been unsuccessful. New modes of ligand binding could greatly contribute to the attempts to develop new drugs that target this enzyme. Here we present a novel binding mode for the biotin carboxylase subunit of ACC with associated ligands. This work is funded in part by National Science Foundation (MCB‐0841134).
Engineering trypsin‐fold serine proteases for inhibitor resistance
Authors: Batt Anna , Baird Teaster
Published: June 2021

In several co‐crystal structures of trypsin and its macromolecular inhibitors, the hydroxyl group of Tyr39 and backbone carbonyl of Phe41 of trypsin lie within hydrogen bonding distance of the P4′ and the P2′ backbone amides of the inhibitor, respectively. Lys60 and Tyr39 form a “cage” around Phe41, likely keeping the backbone carbonyl of Phe41 optimally positioned for the hydrogen bond at P2′. Several trypsinogen variants, in which various residues within the prime‐side binding interface were replaced with a range of alternate amino acid residues, were constructed to evaluate how each of the prime‐side trypsin residues affect macromolecular inhibitor binding. It is hypothesized that disruption of the hydrogen bonds at P2′ and P4′ will allow more conformational freedom and thereby weaken macromolecular inhibitor binding. Initial kinetic characterization and inhibition studies of the variants have been carried out using small commercially available peptide substrates (1–3 residues), and several macromolecular inhibitors. Preliminary data suggest that the constructed variants and wild‐type trypsin show similar affinities for the small peptide substrates (KM = 26 ± 3 μM), while affinity for macromolecular inhibitors appears to vary for the variants relative to wild‐type. This work was supported by NSF CAREER Award MCB‐0643988–02 and NIMHD Award P20MD000544.
Characterization of O‐acetylserine (thiol) lyase from Leucaena leucocephala
Authors: Yafuso Jannai T. , Negi Vishal Singh , Bingham Jon‐Paul , Borthakur Dulal
Published: June 2021

In plants, the final step of cysteine formation involves the addition of sulfide to O‐acetylserine (OAS) and the release of acetate, catlyzed by O‐acetylserine (thiol) lyase (OASTL). The purpose of this study was to isolate and characterize recombinant OASTL from Leucaena leucocephala (leucaena), a protein‐rich legume used as a fodder. Leucaena contains a toxic, nonprotein amino acid, mimosine, which is also produced by OASTL. In order to understand the function of this enzyme, we first studied cysteine synthesis. The cDNA for a cytosolic OASTL was obtained through interspecies subtractive hybridization and Rapid Amplification of cDNA Ends. The sequence was codon optimized and expressed in Escherichia coli under the control of a T7 promoter. A 37‐kDa protein was isolated and used in in vitro enzyme activity assays with Na2S and OAS as substrates where cysteine was detected and quantified using High Performance Liquid Chromatography. The Km was 1.850±0.4146 mM Na2S and the Vmax was 200.6±19.92 μM cysteine minute−1. Central to the mimosine and cysteine biosynthetic pathways is OASTL. Characterization of this enzyme will be essential in the development of a toxin‐free leucaena for its use as a widespread forage crop. This research was funded by NSF Award No. CBET 08–27057.
Fatty acid uptake in the Saccharomyces cerevisiae mutant mon2
Authors: Gasparotto Anthony , Soni Mona , Gasper Brittany , McDonough Virginia
Published: June 2021

The yeast MON2 gene product is a scaffolding protein involved in vesicular budding on the trans‐Golgi membrane. We have found that the mon2Δ mutant shows increased growth sensitivity to undecylenic acid (11:1Δ10), a medium chain unsaturated fatty acid (UFA), compared to wild type cells under the same growth conditions. In order to better understand this effect, we examined wild type and mutant mon2Δ responses in unsupplemented, 18:2Δ9,12, and 11:1Δ10 fed conditions. We have found that when Mon2p is absent, there are altered molecular responses to 11:1Δ10 supplementation, including: small but significant changes in the fatty acid profile; differential localization of BODIPY 500/510 C1, C12, a fluorescent fatty acid analog; decreased expression of OLE1, the gene for the delta‐9‐desaturase; and, altered rates of fatty acid uptake. These changes result in decreased synthesis of endogenous UFAs, which may be responsible for the 11:1Δ10 toxicity. These findings suggest a role for Mon2p in the uptake and trafficking of exogenous medium chain fatty acids, possibly through vesicle transport. Supported by awards from the National Science Foundation‐Research Experiences for Undergraduates and a Research Scholar Award from Howard Hughes Medical Institute.
On‐going Studies on the Structural Stability of E. coli Alkaline Phosphatase by an Undergraduate Biochemistry Laboratory Course
Authors: Hazzard James T. , Schnoebelen Carly
Published: June 2021

Our biochemistry laboratory course engages in a guided‐inquiry exercise in which we study the high degree of structural stability of E. coli alkaline phosphatase (AP) and the roles of two disulfide bonds toward thermal and chemical denaturation. Recently we have compared the denaturation of the enzyme using guanidinium hydrochloride and urea, in the absence and presence of the reducing agent TCEP, using CD and fluorescence spectroscopic techniques. We have also investigated the effect of temperature on the chemical denaturation processes. We also have continued to determine the nature of a chaperonin‐like protein(s) found in the periplasmic lysates that contributes to the dramatic thermal stability of AP, relative to the highly purified commercial enzyme using tandem mass spectrometry. Being a portion of an undergraduate biochemistry laboratory course, these investigations represent data obtained from individual laboratory groups who make oral presentations at the end of their research, providing a more real world approach to the educational experience. This work is supported, in part, by a grant from NSF (DUE 0837398).
Health disparity in Honduras: evaluation of growth status in children ages 6–60 months compared to mother's body mass index
Authors: Endres Emily L , Kemmer Teresa M , Coello Miguel
Published: June 2021

In Honduras, one in four children suffer from malnutrition. The purpose was to determine if growth parameters are associated with socioeconomic status and the mother's body mass index (BMI). Height and weight were obtained from randomly selected children in this cross‐sectional study of 53 children ages 6–60 months and their mothers living in Montaña de la Flor, Honduras. In children, stunted was defined as height‐for‐age <−2 z‐score (HAZ), wasted as weight‐for‐height <−2 z‐score (WHZ), and underweight as weight‐for‐age <−2 z‐score (WAZ). Mother's BMI was defined as underweight <18.5, normal 18.5–24.99, overweight 25–29.99, obese ≥30 and stunting in the mother as height <145 cm. Prevalence of children stunted was 53%; wasted 6%, and underweight 40%. Mother's prevalence was overweight 22%, obese 4% and stunted 30%. Mother's BMI was not significantly associated with children's growth parameters. Of the mother‐child pairs evaluated (n=46) when the mother's BMI was ≥ 25, 15% had a child that was stunted, 13% underweight and none wasted. Significant associations were found between WAZ and telephone ownership, piped water, type of roofing and household food security (child having no breakfast, eating less and going hungry, decreased access to food, and adult going hungry).

Grant Funding Source: USDA NIFA Competitive Grant 2011 67002 30202 and USDA, SD Agricultural Experiment Station Grant SD00H249–08.
Household food security and dietary diversity in the context of an agricultural and market development program in Guatemala
Authors: Cintron Julio R Lopez , Engeseth Nicki J , Dong Faye M , Greene Jennifer C , Wuth Alicia , Peterson Ross , Torres Pablo , Andrade Juan E , Helferich William G
Published: June 2021

WFP's Purchase for Progress (P4P) is a pilot program that enhances agricultural productivity and access to markets among low‐income farmers. We compared food security and dietary diversity between P4P beneficiaries and a control group. We evaluated household conditions, food security (ELCSA), and dietary diversity in 372 households (271 P4P; 101 control) using a cross‐sectional design and mixed‐methods. Most Significant Change methodology was used to characterize participants’ experiences. Education level (EL), number of children (NC), household quality (HQS), food security (FSS), carotenoid‐rich foods (VAS), and dietary diversity for households (HDDS), women (WDDS), children (IDDS), and normalized (HDDSn) were calculated. P4P participants were less food insecure (FSS=7.4±4.4, 9.2±3.1; p<0.01), and had increased dietary diversity (HDDS=8.9±1.8, 7.0±1.8; p<0.01), HDDSn, WDDS, IDDS, and VAS (p<0.01) than controls. For P4P, FSS was associated with EL (r=0.23, p<0.05); and HDDS with EL (r=0.23, p<0.05) and NC (r=−0.17). For controls, FSS was associated (p<0.05) with HDDS (r=0.53), HQS (r=0.61), and NC (r=−0.23). Women empowerment, intra‐household dynamics, higher crop productivity and technical knowledge were dominant themes among P4P beneficiaries. P4P program has a positive effect on food security and dietary diversity, which are perceived as important elements in improving livelihoods.

Grant Funding Source: Ayre international research fellowship (JRL), Hatch (JEA), USDA‐ISE (WGH, NJE, and JEA)
The development of a valid measure of school children's fruit and vegetable consumption
Authors: Taylor Jennifer C , Yon Bethany A , Johnson Rachel K
Published: June 2021

As interventions aim to increase schoolchildren's fruit and vegetable (F/V) consumption, accurate dietary assessment methods not subject to self‐report errors are needed. We tested the validity of three methods in schoolchildren (grades 3–5). F/V consumption was assessed using direct observation (DO)(n=101), digital imaging (DI)(n=165), and digital imaging plus observation (DI+O)(n=167) over four days at two school sites and tested against weighed plate waste (WPW). Validity was tested using Pearson correlations, paired t‐tests, and Bland‐Altman plots. DO (r≥0.92), DI (r≥0.91), and DI+O (r≥0.95) were strongly correlated with WPW for consumption of fruits, vegetables, and F/V combined (p<0.01). Pearson correlations for DI versus DI+O increased for lasagna, soup and whole fruit. Mean biases for total F/V consumption/tray did not differ from WPW when using DO (p=0.39), DI (p=0.56), or DI+O (p=0.38). DO underestimated mean fruit consumption/tray by 3.0 gm (p<0.05), and DI+O overestimated mean vegetable consumption/tray by 2.8 gm (p<0.05). Bland‐Altman limits of agreement for individual F/V consumption were within ~one oz for DO (−35.9 to 38.9 gm), DI (−31.3 to 32.9), and DI+O (−26.8 to 25.0). DO, DI, and DI+O were all valid for assessing mean consumption. DI may be especially promising for assessing mean consumption of fruits, vegetables and F/V combined as excluding observations reduces labor and time.

Grant Funding Source: USDA ‐ Vermont Agricultural Experiment Station and the Bickford Scholar Research Fund