bluuebunny/crossref_metadata_2025 huggingface dataset:
Proteomic assessment of the binding partners of GRK4γ: an approach to understand the molecular mechanisms of GRK4γ action
Authors: Nguyen Jenny H , Liu Jamie , Adams Craig W , Andresen Bradley T
Published: June 2021

The molecular mechanism of action of G protein‐coupled receptor (GPCR) kinase 4γ (GRK4γ) remains largely unknown. In an attempt to better understand how GRK4γ recognizes specific GPCRs and its molecular mechanism of action, we utilized a proteomic approach to identify proteins that interact with GRK4γ. This approach produced dozens of potential interactions. Two interesting interactions were Hsp90 and ubiquitin ligase E3C. Hsp90 is known to interact with other GRKs; to validate this interaction with GRK4γ we co‐immunoprecipitate transfected GRK4γ and endogenous Hsp90. Similar to GRK2, Hsp90 and GRK4γ interact and are disrupted by Geldanamycin (GA), an Hsp90 inhibitor. Moreover, GA decreased the expression of endogenous GRK4γ in a dose and time dependent manner. Additionally, bands that resemble ubiquitinated GRK4γ were observed; to investigate this GRK4γ was immunoprecipitated and blotted for ubiquitin in the presence and absence of proteosome inhibitors. The data clearly show that GRK4γ is ubiquitinated. Together these data indicate that there are a number of proteins that interact with GRK4γ and dynamically regulate the expression of GRK4γ, which may contribute to the function of GRK4γ. This work was partially funded by NSF PFI and Beckman Coulter Foundation grants.
Presence of octopamine and octopamine receptors in ganglia and tissues of Crassostrea virginica
Authors: Welsh Christopher , Bess Fiana , Catapane Edward J. , Carroll Margaret A.
Published: June 2021

Octopamine (OA) a biogenic amine first identified in octopus is well studied in arthropods and gastropods being a neurotransmitter and hormone. OA has rarely been reported in bivalves. Using HPLC and ELISA we showed it present in ganglia and tissues of the oyster Crassostrea virginica, the mussel Mytilus edulis, and clam Mercenaria mercenaria. We found it cardio‐excitatory in oyster and mussel, but cardio‐inhibitory in clam. We localized OA and OA receptors in tissues using immunohistofluorescence. We used pan TAAR 1E antibodies, which react to OA, beta‐phenylethylamine, p‐tyramine and tryptamine receptors, but not classical biogenic amines and histamine receptors, and visualized with FITC conjugated 2E antibodies. Tissues were fixed with paraformaldehyde, treated with 1Eand 2Eantibodies, paraffin embedded, sectioned and viewed with a fluorescence microscope. To detect OA we used anti‐OA 1E antibody (OA conjugated to KLH), and visualized with FITC conjugated 2E antibodies. Tissues were fixed with EDAC treated with 1E and 2E antibodies and paraffin embedded and sectioned or frozen, cryostat sectioned and viewed. The TAAR antibodies revealed OA receptors in cerebral and visceral ganglia, heart, gill, adductor muscle and digestive tract. OA antibodies revealed OA in cerebral and visceral ganglia, heart and blood cells in the gill blood channels. The study shows the presence of OA receptors and OA in ganglia and organs of the oyster. The distribution of the OA fluorescence as well as previous HPLC data suggests it may be a hormone in the animal as it appears to be very wide‐spread. The work was supported by 0516041171of NYSDOE and 0622197 of DUE Program of NSF.
Use of non‐cardiac fibroblasts to create three‐dimensional and functionally active cardiac fibers.
Authors: Karabekian Zaruhi , Jamshidi Aria , Ding Hao , Posnack Nikki G , Sarvazyan Narine
Published: June 2021

Recent studies indicate the necessity of including fibroblast cells in the generation of engineered heart tissue. Our goal was to test if noncardiac fibroblasts can serve as a substitute for cardiac fibroblasts to create macroscopic functional 3D cardiac fibers. Our second goal was to test if downregulation of Major Histocompatibility Complex Class I (MHC‐I) molecules minimizes the immunogenicity of noncardiac fibroblasts. Both NIH3T3 and WT‐19 mouse embryonic fibroblasts were capable of generating 3D‐fibers in Matrigel matrix, albeit with slightly different morphology. An optimal fibroblast‐myocyte ratio and cell concentration was required to create structurally intact and actively beating fibers. In addition, extracellular scaffold density had to be adjusted for each fibroblast cell type. Fibroblasts required non‐lethal irradiation in order to prevent overgrowth of nonreplicating cardiomyocytes. Downregulation of MHC‐I molecules via inhibition of beta‐2‐microglobulin expression increased fibroblasts survival in vitro and in vivo. We conclude that noncardiac embryonic fibroblasts can provide structural support to create functional 3D fibers from embryonic stem cell‐derived cardiomyocytes (supported by the AHA SDG 5680036 & NSF CBET1231549).
Identification of Altered Plasma Proteins by Proteomic Study in Valvular Heart Diseases and the Potential Clinical Significance
Authors: HE Guo‐Wei , GAO Ge , XUAN Chao , BAI Xiao‐Yan , LIU Xiao‐Cheng , JING Wen‐Bin , YANG Qin
Published: June 2021

The genetic basis and proteomics of valvular heart disease (VHD) are important for diagnosis and treatment but little is known about biomarkers in rheumatic (RVD) and degenerative (DVD) VHD. Differential plasma protein analysis identified 18 differentially expressed protein sports and 14 corresponding proteins or polypeptides by two‐dimensional electrophoresis and mass spectrometry in 120 subjects. Two up‐regulated (complement C4A and carbonic anhydrase 1) and three down‐regulated proteins (serotransferrin, alpha‐1‐antichymotrypsin, and vitronectin) were validated by ELISA in enlarging samples. The plasma levels (n=40 in each group) of complement C4A in RVD (715.8±35.6 vs. 594.7±28.2 ng/ml, P=0.009) and carbonic anhydrase 1 (237.70±15.7 vs. 184.7±10.8 U/L, P=0.007) in DVD patients were significantly higher and that of serotransferrin (2.36±0.20 vs. 2.93±0.16 mg/ml, P=0.025) and alpha‐1‐antichymotrypsin (370.0±13.7 vs. 413.0±11.6 μg/ml, P=0.019) in RVD patients were significantly lower than those in controls. The plasma vitronectin in both RVD (P=0.006) and DVD (P=0.01) groups was significantly lower than those in normal controls. These revealed potentially useful biomarkers and possible genetic deficiency in valvular diseases. Supported by NSF 81170148, 2010CB529500, 2009DFB30560, 09ZCZDSF04200, 10JCYBJC26400, 2012‐BH110004, 2011BHKZ001 & 2011BHKY002, and CUHK4789/09, China.
Tissue antioxidant status in hemorrhaged inbred rat strains fed diets for 9 weeks varying in levels of methyl group donors
Authors: Dubick Michael A , Klemcke Harold G , Grubbs Dana L , Barr Johnny L , Rose Rajiv
Published: June 2021

We reported previously that inbred rats fed a diet deficient in methyl group donors (SD) during ages 4–13 weeks, had altered liver antioxidant status compared to rats fed a sufficient (SC) or supplemented (SS) methyl group donor diet. The present study reports similar variables in lung, kidney, small intestine and brain (cortex) from this same study. After feeding the diets for 9 weeks, inbred rats [Dark agouti (DA), and Brown Norway/Medical College of Wisconsin (BN/Mcwi)] were bled 50% of their blood volume and survival time from hemorrhage observed up to 240 min. Tissues were collected at death and frozen at −80°C until assayed for total antioxidant potential, TBARS, reduced glutathione (GSH), nitric oxide (NO), NADH and select antioxidant enzyme activities. No significant differences in these variables were noted in any of the SS fed DA or BN/Mcwi rats compared to the SC groups. DA rats fed the SD diet had higher lung TBARS and lower GSH as well as higher kidney TBARS and lower NADH levels than corresponding SC rats. Also, certain antioxidants were generally higher in lung, kidney and jejunum in BN/Mcwi than DA rats. Taken together, these data support a potential role of lower antioxidant status in DA rats contributing to shorter survival time after hemorrhage (p<0.013) in DA than BN/Mcwi rats for all diets. Supported by US Army Medical Research Materiel Command
Heat stress triggers an antioxidant response in porcine skeletal muscle
Authors: Montilla Sandra Isabel Rosado , Johnson Theresa P. , Pearce Sarah C. , Gardan‐Salmon Delphine , Gabler Nicholas K. , Ross Jason W. , Rhoads Robert P. , Baumgard Lance H. , Lonergan Steven M. , Selsby Joshua T.
Published: June 2021

Heat stress (HS) is a major concern for the swine industry as it negatively impacts pig production efficiency. Also, heat related illnesses remain a serious concern for human health. HS pigs exhibit reduced feed intake and diminished growth rate, still, little is known about the molecular effects of HS on pig skeletal muscle. This study aimed to determine the extent to which HS disrupts redox balance in pig red semitendinosus (STR) muscle. Crossbred gilts were subjected to thermoneutral (TN; 20°C) or heat stress (35°C) conditions for 1, 3 or 7 days. To account for reduced feed intake in HS animals a group of TN animals were pair fed to mimic intake of 7 day HS animals. To assess redox imbalance in STR, nitrotyrosine and MDA, markers of oxidatively modified proteins, were measured by Western blots. Additionally, catalase activity, a major antioxidant enzyme, was measured. Nitrotyrosine was similar between all treatment groups; however, 1 and 3 days of HS decreased MDA content by 25% (P<0.05) in HS pigs compared to TN pigs, but recovered to TN levels after 7 days of HS. Catalase activity increased ~30% after 1 day of HS (P<0.05), though after 3 days of HS catalase activity was similar to TN. These results suggest that there is an initial antioxidant response to HS to alleviate free radical injury, however this response is not sustained. Supported by USDA/NIFA Grant No. 2011–67003‐30007.
Heat shock responses in mice after single administration of quercetin
Authors: Islam Aminul , Abraham Preetha , Deuster Patricia , Chen Yifan
Published: June 2021

We investigated the effects of single dose quercetin (Q) on the responses to heat exposure in mice. Animals were exposed to heat under conscious conditions. Prior to the 2nd heat exposure mice were gavaged with Q (n=34) or vehicle (V, n=27). Tissue samples were collected after final heat exposure. Q‐treated mice had significantly lower plasma IL‐6 and IL‐12 levels, higher GSH/GSSG ratio and SOD, and lower 8‐isoprostane levels in plasma compared to V group. No significant differences in core temperature, blood pressure, and heart rate were observed between Q and V groups during heat exposure. However mice in the Q group that had a relatively high peak Tc during the 1st heat exposure showed a significant trend of having a lower peak Tc during the 2nd heat exposure. Overall, heat treatment groups had higher HSP72 and HSP90 protein levels in liver, heart and gastrocnemius muscle tissues compared to control group that were not exposed to heat (n=10). These results indicate single administration of Q produces anti‐inflammatory and antioxidant effects during moderate heat exposure without modulating HSF1 regulation of heat shock proteins. (supported by USUHS R091EH and ONR N0001411MP20025)
Efficacy of Geranylgeranylacetone for heat stroke recovery in Fischer 344 rats
Authors: Quinn Carrie M , Helwig Bryan G. , Leon Lisa R. , Ward Jermaine A , Duran Rocio M.
Published: June 2021

Heat Shock Protein 70 (HSP70) functions as a molecular chaperone to prevent the misfolding and aggregation of proteins under stressful conditions. Geranylgeranylacetone (GGA) is a pharmacologic inducer of HSP70 in the absence of heat stress, making GGA a potential candidate for post‐heat injury/stroke (HI/S) therapy. We hypothesized that GGA administered 24h post‐HI/S would augment liver HSP70 gene expression and accelerate HI/S recovery in rats. Male F344 rats (277.5 ± 5.3 g) were orally administered GGA (200 mg/kg bw; n=9) or vehicle (Veh; 1mL; n=9) 24h post‐HI/S (Tc,Max, 41.8°C; radiotelemetry). GGA had no effect on thermal load accrued during heat exposure. Liver HSP70 gene expression (qPCR) and biomarkers of liver function (VetScan; ALT, albumin, bile acids, BUN) were examined at 2 and 10d of recovery. Liver HSP70 gene expression was increased ~2000‐fold through 10d of recovery; GGA had no effect on this response. Bile acids, ALT and BUN were similar between HI/S and non‐heated controls on 2 and 10d regardless of treatment. Conversely, HI/S rats treated with GGA showed significantly lower albumin levels compared to non‐heated controls at 2d (4.24±0.07 v 4.78±0.03 U/L, respectively; ANOVA, p<0.01). Our results suggest that GGA administration 24h post‐HI/S is not an efficacious treatment in rats and may delay recovery of liver function. Research supported by MRMC. Author views not official US Army or DoD policy.
Complementing Heat Stroke: Activation and Amplification of the IL‐6 and Complement System during Heat Stroke Recovery in F344 Rats
Authors: Leon Lisa R , Duran Rocio M , Helwig Bryan G.
Published: June 2021

The pathophysiological responses to heat stroke (HS) are thought to be due to a systemic inflammatory response syndrome (SIRS) that is initiated by endotoxin leakage across ischemic gut membranes and high IL‐6 production. The complement system (CS) interacts with IL‐6 to modulate the immune response to sepsis, but its role in HS is unknown. We hypothesized that plasma IL‐6 and the liver CS would show a similar time course of activation in male Fischer 344 rats (262 ± 8 g) during 10 days of HS recovery. Plasma IL‐6 (Multiplex; N=9–12/group) and liver expression of 29 genes in the CS pathway (qPCR; N=3/group) were measured at maximum core temperature (Tc,Max=41.8°C; radiotelemetry), 1, 3, 5 and 10 days of HS recovery. Plasma IL‐6 was elevated from Tc,Max to 3 days of recovery with ~100‐fold increase above controls observed on day 1 (4395 ± 201 vs. 41± 22 pg/ml, respectively; ANOVA, P<0.05 ). Plasma IL‐6 was similar to controls on day 5 and 10 of recovery. Liver CS gene expression (5–11 genes/time point) increased ~2 to 6‐fold above controls on day 1, 3 and 5 of recovery, but peak expression was not observed until day 10 with ~20 to 50‐fold increase above controls (ANOVA, P<0.05). This is the first study to demonstrate liver CS activation in response to HS in rats and suggests that IL‐6‐CS interactions may modulate the SIRS during the early days of HS recovery. Research supported by MRMC. Author views not official US Army or DOD policy.
Early Activation of Liver Apoptotic Signaling Pathways during Heat Stroke Recovery in Mice
Authors: Leon Lisa R , Dineen Shauna M. , Clarke David C.
Published: June 2021

The inflammatory response to heat stroke (HS) often culminates in multi‐organ failure and death. Liver damage is a long‐term HS response but the early signaling events leading to damage are unknown. This study examined liver apoptotic pathway activation during early HS recovery in mice. Liver expression of 15 signaling phosphoproteins (Multiplex; p‐Akt, p‐c‐Jun, p‐CREB, p‐ERK, p‐GSK3, p‐HSP27, p‐IkBα, p‐IRS1, p‐JNK, p‐MEK1, p‐p38MAPK, p‐p53, p‐p70S6K, p‐p90RSK, p‐STAT3) were measured in B6129F2 mice (n=9/gp) at baseline, maximum core temperature (42.4°C), ~1h and ~3h of HS recovery and compared to non‐heated controls. Liver p‐Akt (833.2±112.8 vs. 1611.1±174.0 mean fluorescence intensity [MFI]) and p‐p38 MAPK (6.6±1.4 vs. 16.6±3.1 MFI) were decreased in HS compared to control mice at all time points (ANOVA, P<0.05). Conversely, p‐JNK was increased above controls through ~1h (79.2±17.7 vs. 21.7±3.3 MFI) and p‐c‐Jun was increased at all time points with a peak at hypothermia (2733.6±616.9 vs.153.8±8.2 MFI; ANOVA, P<0.05). p‐STAT3 was elevated in HS mice at ~1h with a peak at ~3h. HS minimally affected all other proteins. Phosphorylation of c‐Jun and JNK with concomitant Akt and p38 MAPK dephosphorylation suggests liver apoptosis is initiated during early HS recovery. STAT3 activation may be a compensatory response to limit liver damage. Research supported by MRMC. Author views not official US Army or DOD policy.
The Association between Pulse Pressure and Stroke Volume during Lower Body Negative Pressure and Graded Hemorrhage
Authors: van Helmond Noud , Johnson Blair D. , Curry Timothy B. , Convertino Victor A. , Joyner Michael J.
Published: June 2021

Due to the relationship between stroke volume (SV) and pulse pressure (PP) during lower body negative pressure (LBNP), we hypothesized that PP reflects SV during hemorrhage (HEM). After a 5‐minute baseline, 6 men were exposed to LBNP at ‐15, ‐30, and ‐45 mmHg for 5 minutes at each pressure. After 45 minutes of recovery, three progressive 333‐ml stages of HEM (1 L total) were performed. SV was assessed using a CCNexfin and PP was calculated using an arterial catheter. SV (111 ± 2.1 vs. 77 ± 6.7 mL; p= 0.04) and PP (58±2.7 vs. 41 ± 4.3 mm Hg; p = 0.03) decreased from baseline to ‐45 mmHg LBNP. SV (118 ± 4.4 to 99 ± 6.8 mL; p = 0.06) and PP decreased (58 ± 2.9 to 48 ± 4.9 mmHg; p = 0.06) from baseline to ‐1 L of HEM. PP correlated with SV during HEM (amalgamated r2= 0.98) and LBNP (amalgamated r2= 0.99). There was limited inter‐subject variability in PP vs. SV during LBNP (r2 range 0.57 to 0.99, 5/6 had an r2 >; 0.90), but substantial inter‐subject variability during HEM (r2range 0.21 to 0.99, 2/6 had an r2 >; 0.90). Reductions in PP induced by LBNP or HEM are strongly correlated with SV. However large inter‐individual differences in this correlation during HEM, which may indicate a significantly variable compensatory response in human arterial distensibility, may limit the use of PP as a surrogate for SV during clinical blood loss. Funded by U.S. Army MRMC Combat Casualty Care Research Program Grant # W81XWH‐11–1‐0823.
Vending machines, sugar‐sweetened beverage (SSB) use, and weight status of college students
Authors: Koenings Mallory , Horacek Tanya , Kattelmann Kendra , Byrd‐Bredbenner Carol , Gurka Matthew , Phillips Beatrice , Olfert Melissa D.
Published: June 2021

This study aimed to identify relationships between above average SSB intake of 1160 college students from 9 campuses, their weight, and prevalence of SSB sold in 67 vending machines on those campuses. SSB were categorized as fruit juice, coffee, and soda. Generalized linear mixed models with random campus effect showed: SSB prevalence was not related to intake. Males were 3 times more likely (CI 2.2–5.0, p<0.01) to drink soda, twice as likely (CI 1.4–2.6, p<0.01) to drink fruit juice, and drank twice as much SSB (CI 1.4–3.7, p<0.01) as females. Compared to non‐Hispanic Whites (NHW), Asians were less likely (OR=0.3, CI 0.1–0.9, p<0.05) to drink soda and non‐Hispanic Blacks (NHB) were 3 times more likely to drink fruit juice (CI 1.6–8.0, p<0.01) and drank 3 times more SSB (CI 1.5–4.5, p<0.01). Total SSB intake was not related to weight status, but compared to NHW, Asians weighed 6.1kg less (CI −0.3–−2.9, p<0.01) and NHB weighed 7.2kg more (CI 3.1–11.3, p<0.01) and had 2.4kg/m2 greater BMI (CI 1.2–3.7, p<0.01). Among college students, males drink more SSB than females; Asians drink less SSB and weigh less; and NHB drink more SSB, weigh more, and have greater BMI than NHW.

Grant Funding Source: USDA Grant 2009–55215‐05460
A Fluid Structure Interaction Model of Bileaflet and Monoleaflet Mechanical Heart Valve Flow Dynamics
Authors: Kimiaghalam Morteza , Rubenstein David Alan , Yin Wei
Published: June 2021

The goal of the study was to develop a 3D numerical model to evaluate the opening and closing behavior of a monoleaflet and a bileaflet mechanical heart valve in a left‐ventricle simulating flow chamber. A diaphragm located at the bottom of the chamber drove the flow in the chamber to circulate through two mechanical heart valves on each side of the chamber. 3D Fluid Structure Interaction (FSI) approaches (ANSYS) were employed to describe the movement of the diaphragm and the valve leaflets. The opening angle of the bileafelt heart valve was set at 85°, and that of the monoleaflet heart valve varied between 45° and 85°. Hemodynamic performance of these valves were evaluated by maximum flow velocity through the valve, average pressure gradient across the valve, valve opening and closing time, and effective orifice area. The results demonstrated that the 3D numerical model was able to accurately simulate the opening and closing of these heart valves. The hemodynamic performance of the monoleaflet heart valve was heavily dependent on the opening angle. An opening angle between 75° to 80° provided the best hemodynamic performance of the monoleaflet heart valve. At this angle, the flow conditions around the monoleaflet valve was slightly better than that around the bileaflet valve, with a large effective orifice area, lower maximum flow velocity, and lower average transvalvular pressure. This work is supported by NSF‐DMR0907291.
Associations of maternal obesity and psychosocial factors with breastfeeding intention, initiation, and duration
Authors: Hauff Laura E. , Leonard Stephanie A. , Rasmussen Kathleen M.
Published: June 2021

Little is known about how psychosocial factors (PSF) influence breastfeeding (BF) behavior of obese women. Our objectives were to determine a) whether maternal prepregnancy body mass index (BMI) is associated with social knowledge of, social influence towards, maternal confidence in, and behavioral beliefs about BF; b) whether these PSF predict BF outcomes; and c) whether prepregnancy BMI is associated with these outcomes, independent of the PSF. Longitudinal data from the Infant Feeding Practices Study II (n=2890) were analyzed using logistic and proportional hazards regression models. Prepregnancy BMI was associated with confidence in (P<0.0001), social influence towards (P=0.04), and social knowledge of BF (P<0.0001), but not with behavioral beliefs in BF (P=0.48). Overweight and obese women did not differ from normal‐BMI women in intention to breastfeed (P=0.06), but they had lower odds of ever BF (P=0.03), and were at greater risk of ceasing exclusive (P=0.0008) and any BF (P=0.02) earlier. Only the association with exclusive BF remained significant (P=0.01) after controlling for PSF. All the PSF were positively associated with each BF outcome. We conclude that women with high prepregnancy BMI have psychosocial characteristics associated with poor BF outcomes, despite having strong intentions to breastfeed. Funded by NIH (5T32HD007331) and USDA (Hatch 399307).

Grant Funding Source: NIH (5T32HD007331) and USDA (Hatch 399307)
Dietary supplementation with white button mushroom augments the protective immune response to Salmonella vaccine in mice
Authors: Wang Junpeng , Niu Xiuli , Du Xiaogang , Smith Donald , Meydani Simin Nikbin , Wu Dayong
Published: June 2021

We previously showed that dietary white button mushrooms (WBM) enhance natural killer cell activity and in vitro WBM promotes maturation and function of dendritic cells (DC). The current study investigated if WBM consumption could enhance pathogen‐specific immune response. C57BL/6 mice were fed diets containing 0, 2, or 5% WBM for 4 wk before oral vaccination with live attenuated Salmonella typhimurium. Four wk after immunization, mice were orally infected with S. typhimurium SL1344. Results showed that immunization increased animal survival while WBM alone had no effect; however, among immunized mice, 2% WBM group had a higher survival rate than other groups. We then fed mice 2% WBM to determine the underlying mechanism. We found that Salmonella‐specific blood IgG and fecal IgA levels were higher in WBM‐fed mice than the control mice. WBM‐fed mice also had higher levels of IgG2a and unchanged IgG1 leading to a higher IgG2a/IgG1 ratio, which indicates an enhanced Th1 response. We further found that at 4 wk post‐immunization, WBM‐fed mice had more DC in spleen, and these DC expressed higher levels of the activation markers CD40 and MHC‐II. Together, these results suggest that WBM may improve the adaptive immune response to Salmonella, a food‐borne pathogen, at least in part through activating DC function. Supported by the Mushroom Council and mushroom industry funding partners, and the USDA contract #58–1950‐0–014.
Interactions of all‐trans retinoic acid and interleukin‐4 in the development of alternatively activated lung macrophages
Authors: Heibel Sandra Kirsch , Chen Celine , Perry Trinity , Urban Joseph F. , Dawson Harry D.
Published: June 2021

Previously, we showed that all‐trans retinoic acid (ATRA), the most active vitamin A (VA) metabolite, increased in vitro polarization of pig alveolar macrophages (AM) toward an AAM phenotype after interleukin‐4 (IL‐4) treatment and increased the expression of IL‐4 in the lungs of pigs infected with the parasite, Ascaris suum. Others have shown that VA may reduce the susceptibility to additional parasitic infections in animals and humans. The mechanism(s) underlying these effects of VA remain undetermined. Herein, we demonstrate that AM, treated in vitro with IL‐4 or isolated from the lungs of A. suum infected pigs, had 2–8‐fold increases in mRNA expression of alcohol and aldehyde dehydrogenases involved in ATRA generation. At the epigenetic level, ATRA increased mRNA expression of cytochrome P450, family 26, subfamily A, polypeptide 1 (CYP26A1), an enzyme involved in ATRA catabolism, through a 7‐fold increase in methylated activator histone H3K4me3 binding to the CYP26A1 promoter. In contrast, IL‐4 pre‐treatment reduced ATRA‐induced CYP26A1 mRNA expression 4‐fold through a 4 fold increase, above control, in the binding of methylated repressor histone H3K27me3 to the promoter. In conclusion, VA, via metabolism to ATRA, and IL‐4, by favoring conditions that lead to increased ATRA, may synergistically sustain an AAM response to Th2‐dependent responses.

Grant Funding Source: USDA/ARS Project Plan 1235–51000‐055–00D.
A pilot study examining the application of plasma alkyresorcinols (AR) and urinary AR metabolites as biomarkers of compliance
Authors: McKeown Nicola M , Marklund Matti , Ma Jiantao , Livingston Kara , Jacques Paul , Blumberg Jeff , Chen C‐Y. Oliver
Published: June 2021

Alkylresorcinols (AR) are phenolic lipids present in the bran of wheat and rye. Plasma AR and their urinary metabolites can be suitable biomarkers for whole‐grain consumption of these cereals. The objective of this study was to examine plasma AR and urinary AR metabolites in response to whole wheat (WW) consumption. In a randomized crossover study, 19 subjects (10M, 9F; BMI 22.0 kg/m2; age 26 y) incorporated either 3 (LWW) or 6 (HWW) servings of WW daily into their regular diet for 1 wk. Subjects completed a 2‐wk washout period, abstaining from all WW consumption, before each intervention. Fasting blood and 24 h urine were collected before and after each intervention. TFAA‐derivatized plasma AR (C:19, 21, 23) were quantified by GC‐MS after diethyl ether and solid phase extractions. Urinary AR metabolites (3,5‐dihydroxybenzoic acid and 3‐(3,5‐dihydroxyphenyl)‐propanoic acid) were determined using HPLC with electrochemical detection after enzymatic deconjugation and ethyl acetate extraction. Urinary total AR metabolites was significant higher after HWW to LWW (63.9 vs. 35.7 μmol, P <0.001). This dose‐response relationship was independent of age, sex, energy intake, and baseline urinary AR metabolites level. Plasma total AR was higher, but not significantly, after HWW to LWW (58.4 vs. 44.5 ng/ml, P=0.5). The results suggest that urinary AR metabolites may be useful as biomarkers of compliance in intervention studies of whole wheat.

Grant Funding Source: the United States Department of Agriculture (USDA agreement No. 58–1950‐7–707) and the General Mills Bell Institute of Health and Nutrition
Hepatic biotransformation of alkylresorcinols is mediated via cytochrome P450 and β‐oxidation.
Authors: Marklund Matti , McKeown Nicola M. , Blumberg Jeffrey B. , Chen C‐Y. Oliver
Published: June 2021

Alkylresorcinols (AR) are phenolic lipids present in the bran of whole grains, particularly in wheat and rye kernels. AR may serve as a biomarker of whole grain intake. While AR pharmacokinetics and principal end metabolites have been reported, the metabolic pathways contributing to their relatively rapid elimination from the circulation remain to be established. We investigated whether hepatic ω‐ and β‐oxidation mediate the catabolism of the AR homolog C19:0 to form 3,5‐dihydroxybenzoic acid and 3‐(3,5‐dihydroxyphenyl)‐1‐propanoic acid (DHPPA) using 3 in vitro platforms: human recombinant cytochrome P450 4F2 (CYP4F2), human liver S9 fraction (which includes microsomal and cytosolic enzymes), and HepG2 cells. After incubation with 10 μmol/L C19:0, one hydroxylated metabolite was formed by CYP4F2 and its production appeared to plateau after 1.0 h. After incubation of 10 μmol/L C19:0 with human S9 for up to 4 h, one hydroxylated and one carboxylated metabolite were identified, with their production increasing linearly with time. When HepG2 cells were treated with 5, 10, or 25 μmol/L C19:0 for 48 h, DHPPA was excreted into the media in a dose‐dependent manner. Our results are consistent with a metabolic pathway of absorbed AR to phenolic acids via CYP4F2‐mediated ω‐hydroxylation. Funded by the General Mills Inc. and USDA.
Tissue distribution of nobiletin and its metabolites in mice after oral administration of nobiletin
Authors: Wang Minqi , Zheng Jinkai , Zhong Zhimei , Song Mingyue , Wu Xian
Published: June 2021

Nobiletin (NBT), a major citrus polymethoxyflavone, has been shown to have various beneficial bioactivities such as anti‐inflammatory and anti‐carcinogenic effects. However, the in vivo absorption, metabolism, and tissue distribution of NBT is poorly understood. In this study, A/J mice were fed NBT (250, 500, or, 1000 ppm in diet) for 16 weeks, and then the tissue levels of NBT and its major metabolites i.e. 3′‐hydroxylnobiletin (N1), 4′‐hydroxylnobiletin (N2) and 3′,4′‐dihydroxylnobiletin (N3) were determined by HPLC analysis. Our results showed that NBT was extensively metabolized in the small intestine to glucuronide and sulfate conjugates of N1, N2 and N3. In contrast, these conjugates were largely deconjugated in the colon presumably by colonic microflora. It was also evident that considerable amount of N1 and N2 were transformed to N3 in the colon. Moreover, different tissues showed distinct patterns of metabolite accumulation. For example, N2 was dominant in the small intestine and liver, while N3 was dominant in the spleen and colon. Very interestingly, NBT showed high accumulation in the brain, which was even much higher than that in the liver. The plasma levels of total NBT and its metabolites were as high as 25μM. Overall, we revealed detailed information on absorption, metabolism, and tissue distribution of NBT in mice, which is very important for further understanding of bioactivities of NBT.

Grant Funding Source: AICR, NIH, NAS and USDA
Association of whole grain intake and longitudinal changes in abdominal adiposity in the Framingham Heart Study
Authors: Wang Huifen , Quatramoni Paula , Fox Caroline S , Jacques Paul F. , McKeown Nicola M.
Published: June 2021

Longitudinal studies on the association of whole grain (WG) intake and abdominal adiposity are scarce. We examined the longitudinal association between WG intake and annualized changes in waist circumference (WC) in the Framingham Heart Study (FHS) Offspring adults. WC was measured approximately every 4 years over a median of 14 years of follow‐up. WG intake (g/d) was estimated by FFQ and calculated with and without added bran and germ. Repeated measures models, with data from 4 exam periods (n=11,372 observations), were used to examine associations between WG intake and annualized changes in WC. After adjustment for time‐varying and invariant demographic and lifestyle factors (including WC at the beginning of each exam interval), greater WG intake was associated with less annualized gain in WC. Adults who consumed more whole grains daily (median intake of 39 g/d) gained less abdominal fat per year (0.63 ± 0.04 cm gain in WC) than those who consumed fewer whole grains (median intake of 4 g/d) (0.79± 0.04 cm gain in WC). The inverse association between WG intake and change in WC was stronger in normal weight adults. The association remained significant after further adjusting for lipid‐lowering and blood pressure medications and diabetes. Based on these findings, a randomized trial is warranted to examine whether incorporating WG foods in the diet helps to mitigate gains in abdominal adiposity over time.

Grant Funding Source: The National Institute of Health/National Heart, Lung, and Blood Institute (Contract N01‐HC‐25195), the United States Department of Agriculture (USDA agreement No. 58–1950‐7–707) and the General Mills Bell Institute of Health and Nutrition
Race, pregravid BMI and gestational weight gain determine placental fatty acid transfer
Authors: Smither Allison Rose , Durham Holiday A. , Buzhardt Sarah , Moore FA , Cefalu William T. , Lammi‐Keefe Carol J.
Published: June 2021

Long chain polyunsaturated fatty acids (LCPUFA), arachidonic acid (AA) and docosoahexaenoic acid (DHA), are essential fatty acids for fetal growth (AA) and development (DHA). The fetus relies on maternal stores and placental transfer for LCPUFAs. In our study population of Hispanic and African‐American women, the relationships between maternal and fetal red blood cell (RBC) AA and DHA were examined and predictors of fetal fatty acids such as age, maternal pregavid body mass index (BMI), gestational weight gain, and race were determined. Maternal and venous cord blood and health histories were obtained at delivery (n=9). Gas chromatography was used to assess RBC fatty acids (wt %). Pearson correlations were used to analyze relationships. Stepwise regression determined the most significant predictors of AA and DHA transfer. There was preferential transfer of DHA and AA from mother to fetus over precursor fatty acids. Maternal DHA was positively correlated with fetal DHA (p=0.046), while AA was not. BMI and gestational weight gain were the strongest predictors of AA transfer to the fetus (p=0.045). DHA was less efficiently transferred in Hispanics than in African‐Americans (p=0.036). Future work is needed to better understand how other factors such as maternal diet affect these findings.

Funded by a Howard Hughes Medical Institute grant to LSU (ARS); LSU AgCenter; USDA‐AFRI (CJL‐K)
Effects Of Modifying Both The Linoleic and Linolenic Acid Content of a 45% Kcal DIO Diet on Body Weight And Metabolic Parameters in C57BL6J Mice
Authors: Ward Robert Edwin , Zhou Albert , Lefevre Michael , Hintze Korry
Published: June 2021

The objective of this study was to test the effects of modifying both the ratio and concentration of linoleic and linolenic acids in a DIO mouse model of metabolic inflammation. B6 mice were fed high‐fat diets (45% kcal) for 18 weeks: AIN‐93G (control); 10% (kcal) n6 1% n3 (HL; 10:1 ratio); 10% n6 5% n3 (HH; 2:1 ratio); 5% n6 1% n3 (LL; 5:1 ratio); 5% n6 5% n3 (LH; 1:1 ratio). Plasma cytokines and metabolic hormones were measured with a multiplexed bead assay. Results indicate the diets did not affect food intake, body mass, liver weight, subcutaneous, gonadal, retroperitoneal and total body fat, fasting glucose, HOMA‐IR, OGTT or plasma LPS. All experimental groups had higher body weight, fat depots, GIP and leptin than control. HH group consumed less food during weeks 3–12 and had lower plasma insulin at the end compared with the other groups. LH group had more mesenteric fat and lower plasma IL10 compared with the other experimental groups. LL group had lower GLP‐1 and MCP‐1 compared with the other 4 groups. Taken together, high n3 PUFA reduced anti‐inflammatory cytokine IL10, low PUFA reduced inflammatory cytokine MCP‐1, high PUFA reduced food intake and plasma insulin. In conclusion, PUFA at 6–15% kcal with different n6:n3 ratios had statistically significant, but seemingly minor effects on metabolic inflammation in a DIO mouse model. This project was funded by a USDA NIFA grant UTA01065.
Using Community Readiness Assessment to Inform Environmental Intervention Development in the Children's Healthy Living (CHL) Program in Remote Underserved Minority Populations of the Pacific
Authors: Fialkowski Marie Kainoa , DeBaryshe Barbara , Areta Ropeti , Belyeu‐Camacho Tayna , Bersamin Andrea , Rojas Gena , Vargo Agnes , Nigg Claudio , Guerrero Rachael Leon , Luick Bret , Novotny Rachel
Published: June 2021

The prevalence of early childhood obesity among Native Hawaiians, Pacific Islanders and Alaska Natives is disproportionately high compared to other US population groups. Sustainable, multi‐strategy, community‐based approaches, rather than individually targeted approaches, are increasingly recognized as effective strategies to obesity prevention. Previous studies, however, have not examined community readiness (CR) as an important facet to obesity prevention interventions. We present results of CR assessments completed in the Children's Healthy Living (CHL) Program that will inform development and implementation of a community‐based intervention to modify the food and physical activity environment to prevent child obesity in 5 jurisdictions: Alaska (AK), American Samoa (AS), Commonwealth of the Northern Mariana Islands (CNMI), Guam (GM) and Hawaii (HI). A survey assessing 6 dimensions of CR was administered to key informants in 4 target communities per jurisdiction. Possible dimension CR scores range from 1 (No Awareness) to 9 (High Level of Community Ownership). Jurisdiction scores ranged from 3 – 4 in AK (n = 23) and CNMI (n = 22), 4 – 5 in HI (n = 24), 5 – 6 in GM (n = 34), and 6 – 7 in AS (n = 39). Results will be used to tailor CHL's community‐based environmental intervention to match each community's level of readiness.

Grant Funding Source: Agriculture and Food Research Initiative Grant no. 2011–68001‐30335 from the USDA National Institute of Food and Agricultural Science Enhancement Coordinated Agricultural Program
Persistent Effects of a Soy Diet in Early Development on Bone in Female Rats
Authors: ZHANG JIAN , LAZARENKO OXANA , BADGER THOMAS , RONIS MARTIN , CHEN JIN‐RAN
Published: June 2021

We have previously reported that feeding soy protein isolate (SPI) diet to pre‐pubertal animals had a beneficial effect on bone accretion. However, it currently remains unclear whether the positive effects of SPI diet on bone in early life will persist to adulthood. In the present study, we fed postnatal day (PND) 24 weanling female rats with SPI diet for 30 days, and on PND 55 we switched SPI diet to a control casein diet until 6 months of age. At age 6 months, rats either underwent ovariectomy (OVX) or sham operation, and thereafter continued on control or SPI‐diets for 1 or 3 weeks. Using peripheral quantitative CT scanning, we demonstrated that total bone mineral content and trabecular bone mineral density (BMD) were significantly increased in 30‐day SPI‐fed rats compared to their controls (p < 0.05). Both long‐term (continuous) SPI (LT‐SPI) and short term (early life only) SPI (ST‐SPI) diet protected against one week of OVX‐associated bone loss. On the other hand, LT‐SPI diet protected loss of total, trabecular and cortical BMD three weeks after OVX, whereas ST‐SPI diet only protected against cortical BMD loss (p < 0.05). These results suggest that ST‐SPI diet has modest protective effects against sex‐steroid deficiency‐induced bone loss in adult females. Supported in part by ARS CRIS #6251–51000‐005–03S.
Maternal high fat diet promotion of mammary tumor risk in adult progeny is associated with early expansion of mammary cancer stem‐like cells and increased maternal oxidative environment
Authors: Montales Maria Theresa E , Melnyk Stepan , Simmen Rosalia CM
Published: June 2021

Many adult chronic diseases might be programmed during early life by maternal nutritional history. Here, we evaluated effects of maternal high fat diet on mammary gland development and tumor formation in adult progeny. Female Wnt‐1 transgenic mice exposed to high fat (HFD, 45% kcal fat) or control Casein (CAS, 17% kcal fat) diets in utero and during lactation were weaned to CAS diet at postnatal day 21 (PND 21). Relative to the CAS group, offspring exposed to maternal HFD had higher body and mammary tissue weights and elevated blood sugar and serum insulin levels at weaning and at PND110 and greater terminal end bud numbers at PND30. Mammary expression of tumor suppressors PTEN and p53 and pro‐apoptosis Bcl‐2 genes were lower in progeny exposed to maternal HFD. Mammary tumor latency was also lower (P=.01) in HFD (115±21 d) than CAS (190±13 d) offspring but tumor incidence and growth did not differ between groups. Epithelial cells from pre‐neoplastic mammary tissues showed expansion of the CD29highCD24+ (basal) and Thy1+CD29highCD24+ (cancer stem cell) subpopulations with maternal HFD exposure. Dams exposed to HFD had lower serum glutathione (GSH/GSSG) and cysteine (cysteine/cystine) oxidative ratios than CAS dams at weaning, but the two groups did not differ in body weights and insulin levels. Thus, maternal high fat diet, by altering early mammary development, may constitute a major risk factor for adult breast cancer.

Grant Funding Source: USDA‐CRIS 6251–51000‐005–004S, Arkansas Breast Cancer Research Program, Arkansas Biosciences Institute
The satiety response to a standard lunch in habitual breakfast eaters and skippers
Authors: Forester Shavawn M. , Witbracht Megan , Horn William , Laugero Kevin , Keim Nancy L.
Published: June 2021

Background

Insulin, Leptin, GLP‐1, and Glucagon are known regulators of food intake and glucose metabolism. Past studies showed a benefit of eating breakfast on satiety but were possibly confounded by disruption of habitual meal patterns. Thus, we conducted a cross‐sectional study of 30 women who were habitual breakfast eaters (Br‐E) or habitual skippers (Br‐S) to assess the hormones Insulin, Leptin, GLP‐1, and Glucagon following a standard lunch meal.

Methods

Br‐E ate their typical breakfast meal between waking and 10am, whereas Br‐S followed their normal Br‐S routine. Both Br‐E and Br‐S did not consume any food or caloric beverage after 10am until the standard lunch was served. Blood was drawn 5 min pre‐lunch and 5, 20, 35, 50, 110 min post‐lunch. A multiplexed sandwich immunoassay (MSD) was used to measure hormones.

Results

Repeated measures ANOVA revealed group differences that were time dependent for Insulin (p=0.02), GLP‐1 (p=0.02), and Gluc (p=0.02). GLP‐1 was higher in Br‐E at 5 (p=0.02) and 110 (p=0.03) min.

Discussion

Clear differences in circulating hormones between Br‐E and Br‐S suggest that the previous breakfast meal influenced the postprandial response, even though all subjects were post absorptive at the start of the protocol. Our data further support the idea that regularly eating breakfast promotes satiety. Funding: USDA CSREES 2009–35215‐05364; USDA Intramural project 5306–51530‐019–00D.

Grant Funding Source: USDA CSREES 2009–35215‐05364; USDA Intramural project 5306–51530‐019–00D
Modulations of sirtuin 1 protein levels by nicotine and β‐ cryptoxanthin can be mediated by miR‐34a in A/J mice lung cancer model
Authors: Iskandar Anita R , You Min , Wang Xiang‐Dong
Published: June 2021

MicroRNAs (miRs) are small endogenous non‐coding RNAs that can regulate gene expression in a post‐transcriptional manner and target important molecules in carcinogenesis. We recently found that nicotine promoted lung tumorigenesis and emphysema in tobacco‐carcinogen (NNK)‐treated male A/J mice. We found that lung levels of sirtuin 1 (SIRT1) protein were decreased in the mice treated with the combination of NNK and nicotine. Moreover, β‐ cryptoxanthin (BCX) supplementation to these mice suppressed both lung tumor and emphysema development, which accompanied by SIRT1 protein restoration to normal levels without sirt1 mRNA alteration. This observation indicated a potential post‐transcriptional regulation of SIRT1. We then conducted real‐time PCR analyses to measure the lung levels of miR‐34a and miR‐217, which have been shown to hinder sirt1 translation. We observed that the levels of miR‐34a were increased in the lungs of the mice treated with the combination of NNK and nicotine, whereas miR‐217 levels were not different among all groups. The levels of miR‐34a in the lungs of the BCX‐supplemented mice were restored to that of the normal levels, indicating an inverse association between the levels of miR‐34a and SIRT1 protein. Our data indicate that the modulation of SIRT1 protein level by nicotine/BCX can be mediated by miR‐34a regulation in the nicotine‐promoted emphysema/lung tumorigenesis in A/J mice.

Grant Funding Source: U.S. Department of Agriculture grant 1950–51000‐064S
Astaxanthin inhibits TGF‐β1‐induced fibrogenic gene expression by inhibiting the activation of Smad3 and KLF‐6 pathways in LX‐2 and primary mouse hepatic stellate cells
Authors: Yang Yue , Lee Ji‐Young
Published: June 2021

Liver fibrosis, a condition defined by excessive accumulation of extracellular matrix, is triggered by the activation of hepatic stellate cells (HSCs). Astaxanthin (ASTX), a xanthophyll carotenoid, has multiple protective properties against nonalcoholic steatohepatitis (NASH). To investigate anti‐fibrogenic functions of ASTX, we used LX‐2, a human HSC cell line, and primary HSCs isolated from C57BL/6J mice in this study. Increases in the production of reactive oxygen species by transforming growth factor β 1(TGF‐β1) or tert‐butyl hydroperoxide was significantly attenuated by 25 μM ASTX in LX‐2 cells. Pretreatment with ASTX significantly decreased TGF‐β1‐induced mRNA abundance of α‐smooth muscle actin (α‐SMA) and collagen 1 α1 (Col1α1) and Western blot confirmed the reduction in α‐SMA expression by ASTX in LX‐2 cells. The nuclear translocation of major transcription factors transducing TGF‐β1 signal, i.e., Smad‐3 and Krueppel‐like factor 6 (KLF‐6), was noticeably inhibited by ASTX in LX‐2 cells. ASTX also significantly repressed the expression of α‐SMA and restored superoxide dismutase 2 expression in activated primary mouse HSCs. In conclusion, our study demonstrates that ASTX exerts anti‐fibrogenic and antioxidant effects in HSCs in vitro by repressing the activation of Smad‐3 and KLF6 by TGF‐β1, suggesting a potential role of ASTX in the prevention of NASH.

Grant Funding Source: USDA AFRI 2012–67018‐19290
Consumption of Different Sources of Omega‐3 Fatty Acids Alters Lipogenic Gene Expression in Growing Rats
Authors: Mock Kaitlin , Tou Janet , Benedito Vagner , Gigliotti Joseph
Published: June 2021

Currently, there is an increased intake of omega‐3 (n‐3) polyunsaturated fatty acids (PUFA) through diet and supplementation. The objective of this study was to determine if the n‐3 PUFA source plays a role in de novo lipid metabolism by altering gene expression. Female Sprague‐Dawley rats were assigned to one of six diets: corn oil (CO), flaxseed oil (FO), krill oil (KO), menhaden oil (MO), salmon oil (SO), or tuna oil (TO). Most n −3 PUFA sources had decreased expression of hepatic lipogenic genes. Stearoyl CoA Desaturase 1 expression decreased in TO (P=0.04) and MO (P=0.008) compared to CO. Fatty Acid Synthase expression decreased in TO (P=0.006), SO (P=0.001), MO (P=0.001), and FO (P=0.01) compared to CO. Sterol Regulatory Element Binding Protein 1c expression decreased in MO (P=0.007) compared to CO. Peroxisome Proliferator‐Activated Receptor Alpha (PPAR α) was used as a measure of hepatic lipolysis, however, no differences were found. There were no significant changes in expression of lipogenic genes in gonadal tissue of n‐3 PUFA groups when compared to CO. In the gonadal tissue, PPARγ increased in FO (P=0.04) compared to CO, and Hormone Sensitive Lipase had no differences in expression compared to CO. These values suggest that n‐3 fatty acids may be altering de novo lipid metabolism by decreasing expression of lipogenic genes in liver tissue, and increasing expression of a lipolytic gene in gonadal tissue.

Grant Funding Source: NIFA #1004489 and WVU Hatch Grant H459