bluuebunny/crossref_metadata_2025 huggingface dataset:
Presence of inhibitory GABA receptors on serotonin neurons in the bivalve mollusc Crassostrea virginica (1059.5)
Authors: Welsh Christopher , Saunders A.C. , Carroll Margaret , Catapane Edward
Published: June 2021

GABA is a major inhibitory neurotransmitter in molluscs and other animals, but has not been well studied in bivalves. In several bivalves, including Crassostrea virginica, beating of gill lateral cilia is controlled by a reciprocal serotonergic‐dopaminergic innervation from their ganglia. Serotonin (HT) increases beating rates, dopamine slows it down. GABA has no direct effects on cilia beating in C. virginica, however, GABA blocked the cilio‐excitation of HT when both are applied to the cerebral ganglia. We hypothesize cerebral and visceral ganglia contain GABA neurons and the cilio‐excitatory HT neurons in the ganglia contain inhibitory GABA receptors. We used immunofluorescence histochemistry to visualize GABA receptors and HT neurons in cerebral and visceral ganglia. Tissues were dissected, snap frozen, sectioned on a cryostat, fixed with paraformaldehyde, treated with blockers, incubated with 1̊ and 2̊ antibodies and viewed with a fluorescence microscope with FITC and Texas Red filters. We found cerebral and visceral ganglia contained GABA neurons and some HT neurons had GABA receptors on their soma. This study confirms our earlier pharmacological studies indicating a role for GABA in this bivalve, working centrally as an inhibitory ganglionic neurotransmitter to inhibit cilio‐excitatory HT neurons that innervate the gill and helps to explain the ganglionic circuitry by which this could be done.

Grant Funding Source: NIH‐2R25GM0600309, NYSED‐0516041071, NSF‐0622197
Oxidative stress and the impact of prenatal chronic hypoxia on ryanodine receptor generated calcium responses in fetal pulmonary arterial myocytes (1089.11)
Authors: Kaushal Shawn , Romero Monica , Osman Noah , Paez Ricardo , Francis Michael , Taylor Mark , Longo Lawrence , Wilson Sean
Published: June 2021

Ryanodine receptors (RyRs) are sensitive to reactive oxygen species in that this stress increases leak of Ca2+ through the channels. Our recent work shows RyR‐generated Ca2+ sparks are aberrant following prenatal chronic hypoxia (CH) in pulmonary arterial myocytes of fetal sheep. Other evidence suggests that the enhanced Ca2+ leak due to oxidative stress can cause loss of Ca2+ from the sarcoplasmic reticulum (SR); our Ca2+ spark data are consistent with such a loss in SR Ca2+ and CH can cause oxidative stress. To address potential Ca2+ loss we quantified 10 mM caffeine (CAF) elicited Ca2+ release from the SR, as RyR activation provides an index of Ca2+ storage. This was performed in pulmonary arterial myocytes of intact arteries using Fluo‐4 and confocal imaging approaches of fetal sheep that lived in normoxia at low altitude (FN) or fetal sheep that lived in CH at (3,801 m) for >100 days (FH). Using customized analysis software, the data show that under control conditions prenatal CH does not impact the area under the curve (AUC) due to CAF. Thus, the changes in spark activity previously observed likely arise from something other than alterations in SR Ca2+ storage. Pretreating PA with 1 mM tert‐butyl H2O2 resulted in a ~ 31% decrease in CAF‐elicited AUC in FN and a 6% increase in FH. These data suggest that sheep exposed to prenatal CH may adapt, which limits decrements in total SR Ca2+ storage in an attempt to preserve SR function.

Grant Funding Source: NSF MRI 0923559, NIH HD‐069746, P01HD031226, R01HD003807, 5P20 MD‐006988, LLUSOM
Gravitational stress and the evolution of relative tail‐length in arboreal snakes (1100.10)
Authors: Sheehy Coleman , Lillywhite Harvey
Published: June 2021

Gravity imposes potentially important constraints on blood circulation in elongate animals during upright posture or climbing. We test the hypothesis that arboreal snakes have longer tails than non‐scansorial species and interpret the results in context of adaptation to gravity stress. Length data for 226 species in 15 snake families were divided among three gravitational habitats (G‐habitats: stenotopically arboreal, eurytopically arboreal/terrestrial, and non‐scansorial) to categorize climbing habits. We analyzed data using conventional statistics and phylogenetically independent contrasts by constructing a composite tree. Mean relative tail length (RTL) increased and mean relative snout‐vent length decreased with increasing arboreality among the three G‐habitats, independent of phylogeny. Mean total body length (TL) was not different between the two arboreal G‐habitats. Snakes with longer RTLs have a larger percentage of elongate blood vessels contained within relatively tight integument of the tail, which theoretically mitigates postural blood pooling. TL may be constrained in adult female arboreal snakes, and this could be driving an evolutionary tradeoff between longer RTL and lower female fecundity. Our results complement previous studies and suggest that elongated RTLs of arboreal snakes function, at least in part, as an adaptive response to gravity stress.

Grant Funding Source: Supported by the U.S. National Science Foundation grant IOS–0926802 to HBL
Thermoregulatory plasticity in Northern Cardinals (1100.13)
Authors: Komer Maria , Corder Keely , Schaeffer Paul
Published: June 2021

Northern cardinals (Cardinalis cardinalis) are able to endure drastic seasonal variations by physiologically adapting to their environment using thermoregulatory techniques. By examining whole‐animal function, we ask whether decreased metabolic activity during winter nights were a result of decreased activity or an induced state of torpor. We measured body weight, percent fat content, metabolic rate (VO2) across a range of temperatures and body temperature (Tb) using data loggers during both summer (August to early September) and winter (December to January) conditions. We hypothesize that: (1) daily winter Tb fluctuations will be depressed compared to summer displaying an induced torpor, and (2) insulation will play a significant role in thermoregulation. We found a slight depression in circadian temperature rhythms for the winter animals compared to summer animals. Metabolic rate measurements did not differ between summer and winter animals. Fat percentage was significantly increased in the winter birds compared to summer. Our findings suggest that thermoregulatory mechanisms are not as fixed as previously thought and while fat deposits may function to increase insulation, they are more likely important for fuel storage. These data identify subtle changes in seasonal thermoregulation are present, and suggest that mechanisms behind these slight alterations in thermoregulatory states should be further assessed.

Grant Funding Source: Supported by NSF # IOS‐1257455 to PJS
Heat shock proteins protect skeletal muscle against frostbite injury (1102.43)
Authors: Mestril Ruben , Batey Jason , Liskutin Tomas , Schweigert Colin
Published: June 2021

Backgound: Frostbite injury results in serious skeletal muscle damage. Potentially the inflammatory response due to frostbite causes local muscle degeneration. Previous studies have shown that the heat shock proteins (hsp) are able to protect against oxidative stress and inflammation.

Results: In the present study we examined the effects of the heat shock protein inducer, 17‐Dimethylaminoethylamino‐17‐demethoxygeldanamycin (17‐DMAG) which was administered within 45 minutes following frostbite injury. Our results show that rat hind‐limb muscles injected with 17‐DMAG following frostbite injury exhibit less inflammatory cell infiltration as compared to control rat hind‐limb muscles. In agreement with this observation, we found that the increased heat shock protein expression results in a decrease in inflammatory cytokine expression. We also found that administration of 17‐DMAG after frostbite injury is able to preserve muscle function. Our results therefore suggest that 17‐DMAG protects skeletal muscle if administered shortly after frostbite injury.

Conclusion: We conclude that compounds such as 17‐DMAG that induce heat shock proteins are able to preserve skeletal muscle function and structure if injected within 45 minutes after frostbite injury. Our studies provide the basis for the development of a potential therapeutic strategy to treat injury caused by frostbite.

Grant Funding Source: This research was supported by an award from the USAMRMC.
Hemodynamic responses to severe heat stress in rats: implications for blood pressure regulation (1104.20)
Authors: Charkoudian Nisha , Quinn Carrie , Duran Rocio , Leon Lisa
Published: June 2021

Increases in heart rate (HR) and cardiac output, and redistribution of blood flow to the periphery are key cardiovascular responses to heat exposure. We previously observed hysteresis in the HR response to heating in rats, whereby HR was higher for a given core temperature (Tc) during cooling/recovery compared to during heating. In the present study, we evaluated whether this hysteresis was associated with dehydration, and whether specific cardiovascular response patterns were associated with more negative outcomes. Tc, HR and arterial pressure were measured via radiotelemetry in conscious male Fischer 344 rats (n=22; 272.4 ± 4.7 g) during exposure to 37°C (heating phase) and during recovery at 20°C (cooling phase). HR increased substantially during heating. At Tc = 39.5°C, HR was 386 ± 5 bpm during the heating phase and 490 ± 14 bpm during the cooling phase (P< 0.001). HR hysteresis in recovery was coincident with substantial dehydration (7‐8%BW), but returned to baseline after ~20 min despite continued dehydration, suggesting these events were not linked. Independent of hysteresis, rats with the worst outcomes demonstrated a drop‐off in HR at end of heating. These findings provide further insight into multiple simultaneous stressors placed on integrative cardiovascular regulation during and immediately following heat stress.

Research supported by USAMRMC. Author views not official US Army or DoD policy.
Polar bear body temperatures and behavior in the changing Arctic summer (1104.25)
Authors: Whiteman John , Harlow Henry , Ben‐David Merav , Durner George
Published: June 2021

In the Arctic summer, extent and duration of sea ice melt is increasing, displacing Alaskan polar bears from ice in productive near‐shore waters to deep‐water ice or to land. In these habitats, seal prey is likely at low availability. Additionally, extensive open water may increase long‐distance swimming. It is unknown how these changes affect metabolic rates and thermoregulation of polar bears. We recorded body temperature (Tb) with loggers implanted adjacent to the abdominal peritoneum, ventral to the linea alba, in 4 bears on deep‐water ice (May–Oct) and 5 bears on shore (Aug–Oct) in the Beaufort Sea. Tb of ice bears declined from a weekly mean of 37.2–37.5°C (May 23–Jun 03) to 34.7–36.6°C (Sep 21–Oct 09). Bears had unexpected, rapid bouts of Tb decline and recovery. In Aug–Oct, ice bears averaged 0.65 bouts per day (95% were 蠄 12 hours) that reached a mean minimum Tb of 33.6°C; shore bears averaged 1.06 bouts per day (94% were 蠄 12 hours) that reached a mean minimum Tb of 28.6°C. The cause of these bouts and why they were more frequent and reached a lower Tb for bears on shore is not clear. Pending analyses contrast Tb to data collected from loggers implanted in the rump; relationships between bouts of low Tb and swimming behavior; and implications of these patterns for polar bears in the changing Arctic summer.

Grant Funding Source: National Science Foundation 0732713
Characterization of changes in muscle afferent response to stretch following inflammation in male and female mice (1128.1)
Authors: Allawala Anusha , Behnke Stephanie , Zaytseva Dasha , Wilkinson Katherine
Published: June 2021

Chronic pain and musculoskeletal disorders are a leading cause of disability and are more likely to affect females than males. Inflammatory factors can alter the activity of muscle sensory afferents and may contribute to the development of chronic pain or other sensory disorders. We hypothesized that muscle afferent population responses to stretch will be altered following systemic inflammation caused by a Lipopolysaccharide injection (7.5 x 10^5 EU/kg; controls 200 µL saline) 18 hrs prior to the experiment. The extensor digitorum longus muscle and deep peroneal nerve from adult male and female C57BL/6 mice were placed into an in vitro bath and muscle afferent activity recorded with a suction electrode in response to a battery of physiological ramp and hold stretches. The muscle afferent response to stretch was altered by inflammation, but the response in females was opposite of that in males (Multivariate ANOVA, drug x sex interaction, p < .05). Inflamed females showed an increase in muscle afferent firing frequency normalized to baseline frequency during both the ramp and hold phases of stretch, while males showed a blunted response in both. This suggests that inflammation alters the muscle sensory information reaching the central nervous system. As we only measured population responses, future studies will determine the proprioceptor and nociceptor responses to inflammation by measuring the responses in identified individual afferents.

Grant Funding Source: Supported by NSF # DBI 1262832 (AA) and a CSUPERB New Investigator Award (KAW).
Ehs‐1 is required for the expulsion step of the Caenorhabditis elegans defecation motor program (1129.5)
Authors: Freeburg Nelson , Peters Maureen
Published: June 2021

The Caenorhabditis elegans digestive motor program occurs at approximately 50 second intervals and consists of three contractions. The first contraction occurs in the posterior body wall muscle and is followed by a contraction of the anterior body wall muscle approximately 4 seconds later. Finally, waste is expelled via contraction of the enteric muscles. The later contractions are controlled by the release of neuropeptides from the intestine, which activate two GABAergic neurons. A genome‐wide RNA interference (RNAi) screen identified EPS‐15 homologous sequence 1 (ehs‐1) as a candidate gene involved in defecation. Ehs‐1 knockdown does not affect the first contraction or the timekeeping mechanisms. However, the expulsion step occurs in only 10.9% percent of cycles. Ehs‐1 has previously been implicated in synaptic vesicle recycling in neurons, suggesting that the expulsion defect is due to defective GABA release from DVB. However, intestine‐specific knockdown of ehs‐1 results in a 92.3% reduction in expulsion, matching the expulsion frequency following systemic knockdown. Our findings suggest that intestinal expression of ehs‐1 is also required for expulsion. We hypothesize that ehs‐1 is needed for recycling of vesicles in both neurons and intestines.

Grant Funding Source: Supported by a National Science Foundation grant: NSF‐RUI0842830
Assessment of organic and heavy metal contaminants as a function of urban development near rivers of Puerto Rico (1143.9)
Authors: Ortiz Ana , Piñero Luis , Rivera Nilsa , Sosa Maria
Published: June 2021

Anthropogenic activities, such as urbanization, manufacture, agriculture and transportation, often lead to contamination of water resources through runoff from sewer systems or illegal dumping. We wish to determine if urbanization near rivers in Puerto Rico results in increased levels of organic and metal contaminants of anthropogenic origin, to then test how these may affect behavior and nervous system function of river fauna. Water samples from two urban rivers with differing hydrological flow characteristics, Rio Piedras and La Plata, and a control non‐urban river, Rio Mameyes, were analyzed with Gas Chromatography‐Mass Spectrometry and Atomic Absorption Spectroscopy. A wide range screening of organic compounds (OC) identified 35 contaminants in the urban rivers, including esters, phthalates, pharmaceuticals and ingredients in personal care products. Dibutyl phthalate, commonly used in plastics and cosmetics, was found in La Plata at a concentration five times higher than the limit allowed by the EPA for drinking water. Copper, chromium and cadmium in the urban rivers are within EPA standards, although copper in the Río Piedras increased consistently from its origin to its lower reach, while remaining constant throughout the length of La Plata. Further experiments to measure other heavy metals and OCs from pharmaceuticals and personal care products are being conducted.

Grant Funding Source: Supported by: NSF HRD‐1137725; NIH NIGMS R25GM061838; NIH NIGMS 5SC3GM084763
Proteomic investigation of human α7‐nicotinic acetylcholine receptor interacting proteins (1147.5)
Authors: Mulcahy Matthew , Lukas Ronald , Barrantes Francisco , Hawrot Edward
Published: June 2021

The α7‐nicotinic acetylcholine receptor (α7‐nAChR) is a ligand‐gated ion channel widely expressed in the mammalian CNS that is associated with numerous physiological functions. α7‐nAChRs are also the principal high‐affinity α‐bungarotoxin (bgtx) binding proteins in the mammalian brain. There is evidence that α7‐nAChRs are involved in a number of protein‐protein signaling cascades. SH‐SY5Y and SH‐EP1 human clonal cell lines have been utilized extensively to characterize nAChRs. SH‐SY5Y cells naturally express α7‐nAChRs. SH‐EP1‐hα7 and SH‐EP1‐hα7‐Ric‐3 cells have been engineered to stably and heterologously express human α7‐nAChRs alone or along with human Resistance to Inhibitors of Cholinesterase 3 (Ric‐3). Bgtx‐sensitive protein complexes were isolated from each cell line using bgtx affinity immobilization. Bound protein was eluted, reduced and alkylated prior to digestion with trypsin in‐solution. The resulting peptides were analyzed with an LTQ‐Orbitrap mass spectrometer and data analyzed using ProteoIQ™. 216 total proteins (1% peptide FDR; 90‐100% probability) that were not present in controls were identified from multiple preparations of the three cell lines. Identified interacting proteins will serve as a foundation for further investigations in human‐derived cells lines and primary human tissue.

Grant Funding Source: Supported by NIH 1R21AG038774, 1S10RR027027, NSF EPS‐1004057 (EH); PIP 2012 and PICT 2011‐0604 (FJB)
Mitochondrial dynamics in the type 2 diabetic heart (1159.6)
Authors: Gonzalez Eileen , Brito Alfonso , Peltekian Lila , Lopez Vanessa , Delgado Daniel , Yamazaki Katrina
Published: June 2021

Type 2 Diabetes (T2D) mitochondrial effects include imbalanced dynamic events of fusion, fission, and autophagy. The flavanol epicatechin has been previously observed to provide cardioprotection in the setting of ischemia‐reperfusion by reducing inflammation and ROS production. Our objectives are to observe the protective effects of epicatechin in the T2D setting via in vitro and in vivo models. In vitro, C2C12 myoblasts were cultured in glucolipotoxicity treatment (GLT) for 3 days in high glucose (~33 mM) and high fatty acids (0.6 mM) media, ± epicatechin. Our in vivo rat model included controls, sham ± epicatechin (1 mg/kg/day), and diabetes ± epicatechin (1 mg/kg/day). T2D was induced in vivo by high‐energy diet (HED) of 20% lard and 10% glucose. Low dose streptozotocin (30 mg/kg) injection was given at four weeks to decrease insulin secretion, after which epicatechin treatment began with maintained HED. In vitro results thus far show GLT average cell counts of 7.54E+04, which show significant increase in cell death compared to control (3.56E+05). Cells maintained in GLT setting with 100 nM epicatechin yielded an average cell count of 1.78E+0, thus demonstrating a reduction in cell death. Western analysis show enhanced mitochondrial proteins with epicatechin treatment compared to diabetic controls. We predict further data analysis will show a positive correlation between epicatechin and the restoration of normal mitochondrial dynamics in the T2D setting. As the incidence of T2D rises, further investigation into possible therapies and treatments is warranted.

Grant Funding Source: Supported by CSU‐LSAMP fund from the National Science Foundation under Grant No. HRD‐0802628
The role of cerebral oxygenation on tolerance to central hypovolemia (1183.12)
Authors: Kay Victoria , Rickards Caroline
Published: June 2021

Background: Tolerance to central hypovolemia varies between individuals, and recent studies have shown that protection of absolute cerebral blood flow is not an underlying mechanism. We hypothesized that subjects with high tolerance (HT) to central hypovolemia maintain cerebral oxygenation(ScO2) at higher levels of lower body negative pressure (LBNP) compared to their low tolerant (LT) counterparts, despite similar reductions in absolute flow. Methods: 15 healthy human subjects (10 male; 5 female) were instrumented for assessment of ScO2 (via near‐infrared spectroscopy, NIRS) and mean middle cerebral artery velocity (MCAv; via transcranial Doppler, TCD). All subjects completed a presyncopal‐limited lower body negative pressure (LBNP) protocol with an onset rate of 3 mmHg/min. Subjects who made it to 蠅80mmHg LBNP were classified as HT, and subjects who made it to 蠄70 mmHg LBNP were classified as LT. Results: The minimum difference in LBNP tolerance between the HT (N=6) and LT (N=9) group was 206 s (LT=1400±104 s vs. HT=2080±65 s; P=0.0003). Up to ‐45 mmHg LBNP, ScO2 was maintained in HT subjects (P蠅0.538), while the LT (N=9) subjects had a progressive decrease in ScO2 (P蠄0.016) from baseline. MCAv decreased from baseline in both HT and LT subjects (P蠄0.022). There was a strong linear relationship between %∆ MCAv and %∆ ScO2 within the LT group (R2=0.98; P=0.013), whereas a weaker association between perfusion and oxygenation (R2=0.53; P=0.271) was observed in the HT group. Conclusion: In support of our hypothesis, higher tolerance to progressive central hypovolemia was associated with the protection of ScO2, despite an early and significant reduction in cerebral blood flow. This may have important clinical implications for the monitoring of cerebral perfusion and oxygenation in trauma patients.

Grant Funding Source: US Army Medical Research and Materiel Command
Nobiletin inhibits colitis‐associated colon carcinogenesis in mice (121.6)
Authors: Wu Xian , Song Mingyue , Zheng Jinkai , Xiao Hang
Published: June 2021

Nobiletin is a major citrus flavonoid that has shown various health benefits. Herein, we investigated the inhibitory effects of nobiletin on colitis‐associated colon carcinogenesis in azoxymethane/Dextran Sulfate Sodium ‐treated mice. The results showed that oral administration of nobiletin significantly decreased colon tumor incidence by 42% and reduced colon tumor size by 37%. Immunohistochemical analysis demonstrated that nobiletin had significant anti‐proliferative and pro‐apoptotic effects in the colonic tumors. ELISA results showed that nobiletin significantly suppressed the expression levels of inflammatory cytokines IL‐1, IL‐6 and TNF‐ α in colonic tissues by 92%, 69% and 51%, respectively. HPLC analysis indicated that oral administration of nobiletin resulted in high levels of metabolites, i.e. 3'‐demethylnobiletin, 4'‐demethylnobiletin, and 3', 4'‐didemethylnobiletin in the colonic tissues. In contrast, the levels of nobiletin in the colon was 5‐10‐fold lower. Cell culture study showed that the three metabolites had much stronger anti‐proliferative and pro‐apoptotic effects on human colon cancer cells than nobiletin. Overall, our results demonstrated that oral administration of nobiletin significantly inhibited colitis‐associated colon carcinogenesis in mice and these effects can be attributed to the colonic metabolites of nobiletin that had potent anti‐carcinogenic effects in the colon.

Grant Funding Source: Supported by NIH, AICR and USDA
Combination ketogenic diet, ketone supplementation, and hyperbaric oxygen therapy inhibits metastatic spread, slows tumor growth, and increases survival time in mice with metastatic cancer (123.7)
Authors: Poff Angela , Ward Nathan , Seyfried Thomas , D'Agostino Dominic
Published: June 2021

Cancers express an energy metabolism characterized by fermentation in the presence of oxygen (Warburg Effect). Mitochondrial dysfunction forces most cancers to require excess glucose for energy and prevents effective ketone metabolism. Abnormal vasculature creates hypoxia which promotes growth and metastasis by activating HIF‐1, further stimulating glycolysis and enhancing the Warburg Effect. The ketogenic diet (KD) is a high fat, low carbohydrate diet that lowers glucose and elevates ketones and slows cancer progression in vivo. Ketones possess inherent anti‐cancer properties, inhibiting cancer cell proliferation and viability in vitro and prolonging survival in vivo. Hyperbaric oxygen therapy (HBOT) increases oxygenation of the tumor, reversing the cancer promoting effects of hypoxia. We have previously shown that KD + HBOT and ketone supplementation slows cancer progression as stand‐alone therapies. We hypothesized that combining these therapies would be a more effective treatment regimen. We tested this combination therapy in the VM‐M3 mouse model of metastatic cancer. Treated mice exhibited decreased tumor growth and metastatic spread to the lungs, kidneys, spleen, adipose, and liver (p<0.05). Treated mice lived 103% longer than controls (p<001). This study suggests that this combination therapy should be further investigated for potential clinical use.

Grant Funding Source: Supported by Office of Naval Research and a charitable donation from Scivation Inc.
A school‐based intervention’s influence on food knowledge, food preference and gross motor skills among preschoolers (252.5)
Authors: Fitzgerald Nurgul , Mueller Carolina , Willems Cynthia , Zhang Youjie
Published: June 2021

Food intake and physical activity are important factors in obesity prevention. Snack foods provide more than 27% of daily energy intake among children, and gross motor skills during preschool years has been linked to physical activity levels in later years. This study examined the impact of a school‐based intervention (All 4 Kids©) on snack food recognition, food preference, and gross motor skills among preschool age children from minority and low‐income families in New Brunswick, NJ. Forty‐five children (3‐5yr; 20 boys and 25 girls; 32 Hispanic, 11 Black, 2 mixed race/ethnicity) participated in the 24‐lesson intervention (implemented 3 times per week). The 30‐45 minute lessons included age‐appropriate songs, dances and games. Weekly information packets were sent to families, and 3 family events were held at the preschool. Pre‐ and post‐program scores were summed for identifying 18 snack foods, identifying the 9 healthy options, and for stated preference for eating the healthy foods. A gross motor skill scale was calculated based on the measured skills. Pre vs. post differences were analyzed using paired samples t‐ and McNemar tests. After (versus before) the program, mean sum scores significantly increased for identifying foods, identifying healthy foods, and for preference for healthy foods (10.6 vs. 14.3, 4.4. vs. 6.6, and 3.2 vs. 4.9, respectively; p<.0001). Mean gross motor skill scale was significantly higher after the program (20.4 vs. 25.3 p<.0001; possible range: 0‐36). The program was perceived positively by the school and families and was sustained as a part of the school curriculum. The results suggest that the program was effective, and similar approaches can be sustained for a school‐wide influence on lifestyle behaviors among children.

Grant Funding Source: Supported by USDA Award No. 2010‐85215‐20662
Atwater factors overestimate the calorie content of walnuts (371.1)
Authors: Baer David , Gebauer Sarah , Novotny Janet
Published: June 2021

Walnuts are a good source of magnesium, a shortfall nutrient, and contain a significant source of alpha‐linolenic acid. Their unique fatty acid profile likely contributes to their beneficial effect on cardiovascular health, with walnuts being the first whole food for which the US FDA permitted a qualified health claim. These health benefits make walnuts an important part of a healthful dietary pattern; however, their perceived high energy density may be a barrier to consumption. Recent research has shown that Atwater calculations overestimate the energy content of other tree nuts by 5‐20%; however accuracy of the Atwater estimated energy content of walnuts has not been investigated. A RCT was conducted in which 18 subjects were fed 2 diets, a base diet (0 g/d walnuts) and the base diet with all foods reduced and 42 g/d of walnut pieces added. After adaptation to both diets, subjects collected all urine and feces voided for 7 to 10 d at the end of each 3‐wk diet period. These samples and diets were freeze dried and gross energy was measured by bomb calorimetry. The metabolizable energy (ME) content of walnuts was found to be 5.22 kcal/g (146 kcal/serv) as compared to the Atwater‐calculated amount of 6.61 kcal/g (185 kcal/serv). Thus, Atwater factors overestimate by 21% the ME content of walnuts. These data will better inform consumers about the energy content of their foods.

Grant Funding Source: USDA and California Walnut Commission
Associations between parental feeding practices and BMI in middle childhood: the role of children's inhibitory control (379.5)
Authors: Sutter Carolyn , Ontai Lenna , Scherr Rachel , Linnell Jessica , Nicholson Yvonne , Spezzano Theresa , Zidenberg‐Cherr Sheri
Published: June 2021

Parent feeding practices are key to development of dietary habits and childhood obesity, but the role of child characteristics in this association is less clear. Recently, children’s inhibitory control (IC) has been related to eating habits and weight. The objective of the current study was to examine associations between parent feeding practices, IC, and BMI percentile with 267 fourth grade students and their parents. It was hypothesized that IC would moderate associations between parent feeding practices and BMI. Parent feeding practices were collected with the Child Feeding Questionnaire (Birch et al., 2001) (Restriction, Pressure to Eat, and Monitoring). IC was collected with the Children’s Behavior Questionnaire (Rothbart et al., 2001). BMI percentile was calculated from height and weight, adjusted for age and gender. OLS multiple regression results suggest a significant interaction between Pressure to Eat and IC as predictors of BMI percentile (β=3.23, p<.05). Simple slopes analyses indicate Pressure to Eat is related to lower BMI percentile only for children low in IC. Findings suggest that using pressure to influence a child’s eating habits may be beneficial if the child’s low IC interferes with their own ability to regulate their diet.

Grant Funding Source: Supported by UCANR and USDA 2011‐38420‐20082
Eicosapentaenoic acid reduces high‐fat diet‐induced insulin resistance by altering adipose tissue glycolytic and inflammatory function (383.8)
Authors: LeMieux Monique , Nishan S Kalupahana Nishan , Aljawadi Arwa , Pahlavani Mandana , Claycombe Kate , Moustaid‐Moussa Naima
Published: June 2021

We previously reported Eicosapentaenoic Acid (EPA)'s ability to prevent high‐fat (HF) diet‐induced obesity, insulin resistance, and inflammation. In this study, we dissected mechanisms mediating anti‐inflammatory and anti‐lipogenic actions of EPA, using histology/ immunohistochemistry, transcriptomic and energetic analyses of adipose tissues from HF and HF‐EPA fed mice, or 3T3‐L1 adipocytes treated with or without EPA. Microarray and real time PCR analyses of adipose tissue from these mice showed that EPA decreased expression of genes associated with cell proliferation, apoptosis, adipogenesis and glucose metabolism (namely Fasn, Srebf1, Cebpa, Ncoa2, and Axin1). Preliminary extracellular flux studies indicate changes in glycolysis in differentiated 3T3‐L1 adipocytes treated with EPA, compared to those treated with palmitic acid (PA) alone or with EPA. These results are consistent with increased fatty acid oxidation‐related metabolites in gonadal fat after EPA supplementation. Histology and immunohistochemistry results showed a significantly lower mean adipocyte size and macrophage infiltration in mice fed a HF‐EPA diet vs. HF, indicating that EPA prevented HF‐diet induced adipocyte inflammation and hypertrophy. In conclusion, our studies demonstrate that EPA ameliorates glucose homeostasis at least in part by reducing adipose tissue inflammation, glycolysis and lipid accumulation.

Grant Funding Source: USDA, AHA and TTU (COHS and OVPR)
Contribution of sandwiches to USDA food patterns components, What We Eat In America, NHANES 2009‐2010 (393.5)
Authors: Hoy M , Sebastian Rhonda , Enns Cecilia , Goldman Joseph , Moshfegh Alanna
Published: June 2021

Consumption of sandwiches is commonplace in the American diet, but little is known about how they contribute to overall intake of recommended food groups. To describe the contribution of sandwich intake to the USDA Food Patterns (FP) components, one day of dietary intake from individuals age 1+ year (N= 9,317) in What We Eat In America, NHANES 2009‐2010 was analyzed using the Food Patterns Equivalents Database (FPED) 2009‐10. The FPED separates foods and beverages in the Food and Nutrient Database for Dietary Studies 5.0 into 9 FP main components and 28 subcomponents. On a given day, 49% of individuals age 1+ year reported consuming a sandwich (M: 54%; F: 44%). Sandwiches provided 12% of the energy intake of the population, and contributed substantially to total intake of Protein Foods (23%), Grains (20%), Dairy (12%), and Vegetables (8%). In addition, sandwiches provided 15% each of Oils and Solid Fats. Analysis of FP subcomponents showed that sandwiches accounted for 60% of Cured Meat and 28% of Meat, Poultry, Seafood intakes overall and 29% of Cheese intake. They contributed 17% to Whole Grains and 20% to Refined Grains intakes. Sandwiches also provided 14% of the intake of Tomatoes and 13% of Other Vegetables (such as iceberg lettuce, onions, cucumbers, and peppers). These results can inform nutrition education initiatives and consumer messages aimed at promoting adherence to the Dietary Guidelines for Americans.

Grant Funding Source: Supported by USDA Agricultural Research Service
Development of a standard methodology for assessing the satiating effect of foods (47.1)
Authors: Raatz Susan , Vickers Zata , Strubeck Eric , Johnson LuAnn
Published: June 2021

No standard methodology is currently utilized for assessing the relative satiating value of food items. Our goal was to evaluate the validity and reliability of satiety responses in order to develop a standardized methodology for determining the relative satiating capacity of specific food items. A pilot study evaluating the repeatability and correlation of subjective visual analog scale (VAS) responses of hunger and fullness to objective measures of satiety was completed in healthy men and women (n=10). Participants repeated 3 identical testing sessions separated by 蠅1 wk. After a 10 hour fast, time 0 assessments were obtained, followed by consumption of a mixed macro‐nutrient food (340 kcal); additional VAS and blood samples for insulin, glucose and ghrelin were obtained at 30 minute intervals through 240 min. Total areas under the curve (tAUC) for all responses were determined. Reliability of the model was assessed by intraclass correlation coefficient (ICC). Mixed‐model analysis of variance was used to estimate the within‐subject correlation of hunger and fullness responses to glucose, insulin and ghrelin concentrations across all time points. For all measures, ICC was > 0.5 (p蠄 .05). Within subject responses of hunger and fullness, respectively, were correlated to ghrelin, insulin, and glucose. Our data suggest that the model developed for assessing the satiating value of foods is valid and repeatable.

Grant Funding Source: USDA, ARS 5450‐51000‐049‐00D
Evolution of a cross‐institutional asynchronous online 500 level college histology course with interactive lectures and virtual lab component (530.1)
Authors: Ogilvie Robert , Sawyer Roger , Greenwold Matthew , Bao Weier , Thompson Jordan
Published: June 2021

Prior to Spring Semester 2011, the University of South Carolina Biological Sciences curriculum offered only a face‐to‐face histology course where students were required to attend lectures and labs utilizing the microscope. Laboratory space and equipment limited enrollment in the face‐to‐face class which never exceeded 50 consisting mainly of pre‐medical and pre‐dental students. In Spring, 2011, for the first time, we offered a fully online histology course in which all lectures, labs, quizzes and exams were online with no face‐to‐face meetings. Enrollment jumped to 121 in that first semester to include students from more than ten curricular programs across the institution such as anthropology, biochemistry, biomedical sciences and experimental psychology. We now have eight semesters of experience with the course that includes nearly 800 undergraduate, graduate and special certificate program students having completed the course successfully. Lectures have evolved to include SCORM grading of quizzes at the end of each lecture permitting the assignment of viewing lectures as course activity. For the virtual labs, a web based virtual microscopy program is used that contains a collection of over 100 annotated specimens, including two forms of self‐assessment. Recently, we have implemented Biolucida Cloud (MBFbioscience.com) allowing the inclusion of virtual slides in practical exams as unknowns and delivered in random order integrated with Blackboard. Exams have evolved from two attempts with feedback to one attempt with no feedback except the score. Included will be the impact of midterm and final exam construction and administration changes on final grade distribution. The online histology course is now offered year around and remains very popular.

Grant Funding Source: Supported by NSF Grant‐1317771
A revolutionary chemistry curriculum based on fundamental principles of chemical structure and reactivity that integrate organic and inorganic chemistry with biochemistry (530.3)
Authors: Jakubowski Henry , Graham Kate , Johnson Brian , Jones Nicholas , McIntee Ed , Peterson Alicia , Schaller Chris
Published: June 2021

Traditional first year chemistry curricula have remained essentially unchanged for decades although recent movement to an atoms or organic chemistry “first” approach or to introductory biochemistry courses that replace a semester of organic chemistry course have emerged. Outside agencies (American Chemical Society, ASBMB, and the American Association of Medical Colleges) have recently developed guidelines that encourage science departments to develop innovative approaches to content selection and delivery, where competencies and skills replace specified courses. We have embraced these calls and introduced a potentially revolutionary approach to the teaching of chemistry. The unifying principle of our first four courses are based on the structure/reactivity of molecules which “don’t know their classification as organic, inorganic and biochemical structures”. A brief overview of our entire curriculum (content and pedagogy) as well as a more detailed description of our first year courses, Introduction to Chemical Structure and Properties and Introduction to Nucleophiles and Electrophiles (which integrates organic and inorganic with biochemistry) will be presented.

Grant Funding Source: Suporting by NSF
Variants of an active site histidine in Rv0045c esterase from M. tuberculosis (580.3)
Authors: Lukowski Jessica , McKary Magy , Hoops Geoffrey , Johnson R. Jeremy
Published: June 2021

Mycobacterium tuberculosis, from which the Rv0045c protein is isolated, is the most causative bacterium leading to tuberculosis. Histidine (H187), within the active site of this esterase, was the residue of interest in this project. The histidine was mutated to ten different amino acid residues (aspartate, asparagine, glycine, lysine, phenylalanine, proline, serine, tryptophan, tyrosine, and valine) and the catalytic efficiency of the enzyme variants was determined. Variant proteins were produced through site‐directed mutagenesis, overexpression in E. coli cells, and purification using a Ni2+ affinity resin. Evidence from differential scanning fluorimetry (DSF) revealed no significant differences in the thermal stability of the variant proteins. Michaelis‐Menton kinetics revealed that most of the variant proteins showed no difference in catalytic activity from the wildtype protein. The tyrosine variant increased the protein’s overall kinetic efficiency. A sequence alignment with esterases from similar organisms showed that a tyrosine residue is almost as common as the histidine residue present in Rv0045c.

Grant Funding Source: National Science Foundation
The rate‐limiting steps in radical‐mediated carbon‐sulfur bond formation catalyzed by biotin synthase (580.6)
Authors: Jarrett Joseph , Judd Matthew , Fugate Corey , Farrar Christine
Published: June 2021

Biotin synthase is an S‐adenosylmethionine (AdoMet or SAM) radical enzyme that catalyzes the substitution of sulfur for hydrogen in dethiobiotin, generating two new C‐S bonds in the thiophane ring of biotin. In a reaction shared with other members of the Radical SAM superfamily, biotin synthase uses a [4Fe‐4S] cluster to catalyze the reduction of SAM to methionine and a 5'‐deoxyadenosyl radical. This highly reactive radical abstracts a hydrogen atom from the C9 position of dethiobiotin, presumably generating a dethiobiotinyl radical, that is then quenched by a sulfide from a nearby [2Fe‐2S] cluster. Following exchange methionine and 5'‐deoxyadenosine for a second equivalent of SAM, a second reaction sequence focused on the C6 position completes the formation of the thiophane ring. Kinetic analysis of pre‐steady‐state enzyme turnover indicates that formation of the monothiolated intermediate, 9‐mercaptodethiobiotin, is significantly slower than the subsequent ring closure. Using kinetic isotope effect (KIE) analysis, we demonstrate that hydrogen atom abstraction from the C9 methyl group is the overall rate‐limiting step, with a burst phase KIE of ~35 ‐ 40. During in vitro steady‐state turnover, regeneration of the active enzyme through FeS cluster assembly becomes partially rate‐limiting, resulting in a partial masking of this isotope effect to a yield a KIE of ~5. Finally, feeding studies in E. coli with labeled dethiobiotin exhibit a large KIE, suggesting that FeS cluster assembly is not rate‐limiting in the presence of the ISC iron‐sulfur cluster assembly systems. The implications of the very large non‐classical isotope effect will be discussed in terms of enzyme evolution and the potential biotechnological applications for AdoMet radical enzymes.

Grant Funding Source: Supported by NSF (MCB‐1244632 to JTJ)
Protein N‐terminal N‐myristoylation at the proteome scale (587.1)
Authors: Meinnel Thierry , Castrec Benoit , Traverso Jose , Majeran Wojtek , Giglione Carmela
Published: June 2021

N‐Myristoylation (MYR) is a N‐terminal modification, playing a major role in signal transduction pathways. MYR consists in the addition of a myristate to an N‐terminal Glycine, catalyzed by an N‐myristoyl transferase. MYR often acts together with another lipidation located on Cys residues in the close vicinity, S‐palmytoylation. Such lipidations prove most difficult to evidence even with state‐of‐the‐art proteomics approaches and instruments. Although less than 8 residues are enough to define the selectivity of the MYR, the sequence space diversity of MYR is huge (>109). So far, even fine bioinformatics approaches prove limiting to predict this modification. In this context, we have started a comprehensive analysis of this N‐terminal acylation in A. thaliana. With a new high‐throughput in vitro assay and the systematic measurement of catalytic efficiencies, >1200 synthetic octapeptides encoding the N‐terminal part of verified open‐reading frames could be assayed out of the ~2500 possible candidates. It is now possible to better cover the extent of the Myristoylated proteome from A. thaliana using computer modeling and to translate this knowledge to other proteomes including humans. Unexpected new families including thioredoxins for instance could be evidenced. We conclude that MYR is a more frequent modification than initially expected and that its specific recognition motif is extremely complex.

Grant Funding Source: CNRS, ANR, ARC
1H‐NMR metabolomic biomarkers of poor outcome after hemorrhagic shock are absent in hibernators (590.1)
Authors: Bogren Lori , Murphy Carl , Johnston Erin , Serkova Natalie , Drew Kelly
Published: June 2021

Hemorrhagic shock (HS) following trauma is a leading cause of death among persons under the age of 40. During HS, systemic ischemia is followed by reperfusion during medical intervention (I/R) resulting in a disruption of cellular metabolic processes and ultimate tissue and organ dysfunction. Resistance to I/R injury is a characteristic of hibernating mammals. The present study examined if arctic ground squirrels (AGS) are protected from warm HS‐induced global I/R injury due to the ability to maintain a stable metabolic homeostasis during HS. Rats and AGS were subject to HS by withdrawing blood to maintain a MAP of 35 mmHg for 20 min before reperfusion. Body temperature was maintained at 37±0.5°C. Plasma samples were taken immediately before hemorrhage and three hours after reperfusion and were then analyzed via 1H‐NMR spectroscopy for hydrophilic and lipophilic metabolites. Rats had a qualitative shift in their hydrophilic metabolic fingerprint and had alterations in several metabolites (acetate, alanine, glutamine, histidine, β‐hydroxybutyrate, lactate, lysine, and tyrosine ) during I/R indicative of metabolic shut‐down and organ damage. In contrast, AGS, regardless of season, were able to maintain a qualitative 1H‐NMR metabolic homeostasis with little (winter) or no (summer) changes in quantified metabolites during HS‐induced global I/R. In conclusion, AGS are resistant to disruptions in their metabolic processes and organ damage during warm HS‐ I/R.

Grant Funding Source: Supported by USAMRMC W81XWH009‐2‐0134
Prevalence of young child overweight and obesity in the U.S.‐affiliated Pacific region: a meta analysis from the Children's Healthy Living Program (621.6)
Authors: Novotny Rachel , Fialkowski Marie , Li Fenfang , Vargo Donald , Paulino Yvette , Coleman Patricia , Bersamin Andrea , Nigg Claudio , Leslie Jodi , Leon Guerrero Rachael , Deenik Jonathan , Kim Jang , Wilkens Lynne
Published: June 2021

A meta‐analysis of published literature and publicly available agency reports was conducted to estimate overweight and obesity prevalence of 2‐8 year (y) old US Affiliated Pacific (USAP) Region (USAP Islands, Hawai`i and Alaska) children. Contiguous US data served as reference. The search was limited to resources in English and 2‐8y on USAP children data from 2000 and newer and used CDC 2010 overweight and obesity reference data. USAP region sources (n=23) and NHANES articles (n=3; the contiguous U.S reference) were included. Data were disaggregated into single years of age. The children measured for the age group were divided equally and the prevalence was assigned to each individual age. A mixed model regressed the prevalence on a polynomial in age, accounting for the prevalence variance and another model compared jurisdictions. Overweight and obesity increased from 21.0% at 2y, to 39.2% at 8y (p<0.0001) for the USAP. In comparison, combined overweight and obesity increased from 24.2% at 2y to 34.8% at 8y (p<.0001) for NHANES. USAP data showed with a sharp increase in prevalence at 5y. Obesity alone increased from 10.2% at 2y, doubling to 23.6% at 8y (p<0.0001) whereas overweight prevalence was stable from 2y (12.6%) to 8y (14.6%). Jurisdiction differences were dramatic. Further examination of USAP young child obesity is needed.

Grant Funding Source: Supported by USDA Grant 2011‐68001‐30335