bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Vitamin A Deficiency on Retinoid and Energy Metabolism in Liver and White Adipose Tissue
Authors: Brun Pierre‐Jacques , McGrane Mary
Published: June 2021

The liver and white adipose tissue (WAT) play important roles in the metabolism, storage and transport of retinoids. We assessed the effect of vitamin A on expression of genes of retinoid and related metabolism in liver and WAT. Real Time‐PCR was used to assess a panel of genes in vitamin A sufficient (VAS) versus vitamin A deficient (VAD) mice at 9 weeks and 30 weeks of age. Crbp1, Crabp1 and Rbp4, genes involved in retinoid transport, were upregulated in VAD liver but were down regulated in WAT at 9 and 30 weeks. Lrat, Cyp26a1 and b1 genes were repressed in VAD liver and WAT. In liver, the Pparα gene was repressed in VAD; whereas, Pparγ gene expression was not changed at 9 weeks but was downregulated at 30 weeks. In WAT, Pparγ gene expression was repressed in VAD and Pparα gene expression was down regulated at 30 weeks. Over time, there is a decrease in fat mass in VAD which may be caused by a decrease in WAT differentiation as seen by our measured decrease in Pparγ and C/ebp‐α expression in VAD WAT. Leptin and adiponectin are decreased at the mRNA and protein level in VAD WAT. Furthermore, the decreased expression of Lrat and Rbp4 in VAD WAT are markers of its lost capacity to store and secrete retinol. Finally, the combination of decreased fat mass, fatty liver, glucose intolerance and changes in plasma adipokines that we measure in VAD mice, are often associated with lipodystrophy, a condition with severe metabolic outcomes.

USDA Grant #2003‐00883
Green tea and cinnamon polyphenols attenuate mitochondrial dysfunction and glial swelling in ischemic injury
Authors: Panickar Kiran , Polansky Marilyn , Anderson Richard
Published: June 2021

Brain edema is a major complication associated with ischemic stroke and is characterized by a volumetric enlargement of the brain. Astrocyte swelling is a major component of brain edema. We investigated the protective effects of insulin‐potentiating polyphenols isolated from green tea and cinnamon in C6 glial cultures subjected to 5 hr oxygen‐glucose deprivation (OGD). Cell volume was determined using the 3‐OMG method. Ninety minutes after the end of OGD, cell volume increased by 35% when compared with controls which was blocked by polyphenols from green tea (0.1 mg/ml) and cinnamon (0.1 mg/ml) as well as by insulin (10 μM). Individual components of either green tea (EGCG, caffeine, catechin and epicatechin) or cinnamon (dimer, coumarin) did not protect. Likewise, resveratrol (10–100 μM), a polyphenolic anti‐oxidant found generally in grapes and blueberries, did not prevent swelling. Mitochondrial dysfunction, a key feature of ischemic injury, plays an important role in glial swelling. Depolarization of the mitochondrial membrane potential (MMP) was assessed immediately at the end of OGD using TMRE, a fluorescent dye. OGD‐induced depolarization of MMP was attenuated by both green tea and cinnamon. One possible mechanism by which extracts from tea and cinnamon attenuate glial swelling may be through preventing mitochondrial dysfunction. (Supported by USDA & Diabetes Action Research and Education Foundation)
Pomegranate and its polyphenols downregulate gene expression of androgen synthesizing enzymes in human prostate cancer cells overexpressing the androgen receptor
Authors: Hong Mee Young , Seeram Navindra P. , Heber David
Published: June 2021

Early stage prostate cancer (PCa) is treatable, but recurrence after primary treatment with development of androgen‐independent (AI) PCa metastases can lead to death. Therefore, prevention of the development of AI PCa is critical to reducing overall prostate cancer mortality. Cellular adaptations during development of AI include increased androgen receptor (AR) expression and increased intracellular androgen synthesis accounting for resistance to castration in AI PCa. In this study, we examined the effects of Pomegranate (POM) extract and a whole POM juice extract on the expression of genes for key androgen synthesizing enzymes and AR. Expression of the HSD3B2, AKR1C3 and SRD5A1 genes for androgen synthesizing enzymes were determined by real time PCR in LNCaP, LNCaP‐AR, and DU‐145 human prostate cancer cells in vitro. Pomegranate and its polyphenols decreased tumor cell proliferation (P<0.01) in all three cell types. Pomegranate polyphenols also inhibited expression of genes for androgen synthesizing enzymes and AR more than two‐fold most consistently in the androgen independent LNCaP‐AR cell line (P<0.05). Therefore, the effects of pomegranate polyphenols may be mediated in part by inhibition of androgen synthesizing enzymes in PCa when AR is upregulated in AI PCa. Supported by UCLA CNRU CA 42710 and DOD W81XWH‐07‐1‐0158.
Identification of an Orphan 7TM Receptor PSGR as a Functional Intracellular GPCR
Authors: Cheng Huaiyan , Soojhawon Iswarduth , Qiu Rongde , Wang Mi , Sun Chen , Srivastava Shiv , Feng Ying‐Hong
Published: June 2021

PSGR, a prostate‐specific orphan GPCR of olfactory family, expresses exclusively in the endoplasmic reticulum (ER) and Golgi network in surrogate cells including prostate cancer cells and normal human prostate epithelial cells. To determine if PSGR is functional or eventually goes to the cell surface we show here that removal of the N‐ and/or C‐terminal domains of PSGR, fusion of a signal peptide of EGF receptor to the N‐terminal of PSGR and/or tagging with various ER export motifs at the C‐terminal of PSGR failed to express the receptor on the cell surface unless they were coexpressed together with RPT1s, Ric8B, and Gαolf. Structural analysis revealed no obvious ER‐retention motif or any other structural determinants responsible for intracellular expression of PSGR. Surprisingly, PSGR inhibited p53 phosphorylation, but promoted Erk1/2 phosphorylation. Overexpression of the PSGR wild type, but not the mutant (D120A/R121L) incapable of coupling to G proteins and mutants with altered intracellular localization in CHO cells, promoted cell proliferation. These results suggest that PSGR is a functional intracellular GPCR and intact G protein signaling machineries are present in the intracellular organelles such as ER and Golgi. This work also suggest that PSGR may be constitutively active or the cognate ligand is likely intracellular and/or membrane permeable. Supported by USUHS startup funds to YHF
Distribution and expression of Aquaporin HC1 in tissues of Hyla chrysoscelis under different physiological conditions
Authors: Pandey Ram Naresh , Yaganti Sushmita , Frisbie James , Goldstein David L.
Published: June 2021

HC1 is an AQP1‐like water channel protein expressed by the treefrog Hyla chrysoscelis. Expression of HC1 mRNA is widespread among tissues. We used immunohistochemistry to localize HC1 protein within those tissues, and to explore variation in that expression in control (hydrated, 23C), dehydrated (20% loss of standard body mass over one week at 23C), and frozen (−2.5C for 24 hr) animals. Tissue sections were exposed to anti‐HC1 antibody and stained with Texas Red or Cy2 conjugated secondary antibody. In kidney, HC1 was detected in the parietal layer of the glomerulus and in overlying connective tissue. HC1 was expressed in glandular membrane in stratum spinosum of skin, in septa, sinusoids, and acini of liver, and in the gastrointestinal tract in gastric pits and in the lumen of crypts of small and large intestine. We also detected expression in the pulmonary epithelia and in blood vessels of several organs, including skeletal muscle and intestines. Our findings are largely consistent with the role of HC1 as a widely expressed “housekeeper” aquaporin. However, we also have preliminary evidence for some physiological regulation of expression, including down‐regulation in frozen liver and enhanced expression in skin from frozen and dehydrated animals. Research supported by NSF IOB‐0517301.
Growth within an instar reduces jumping performance in American locusts (Schistocera americana)
Authors: Duffy Bridget Mary , Harrison Jon F. , Kirkton Scott D.
Published: June 2021

In developing vertebrates, morphological changes that alter behavior, ecological patterns, and physiological processes are well known. However, the effect of ontogenetic structural changes on insect life‐history are not well understood. During development, American locust grasshoppers (Schistocerca americana) exhibit dramatic increases in body size both across and within instars. We have shown previously that as grasshoppers develop across instars, older animals have reduced jump endurance and increased lactic acid production in the jumping muscle. Within an instar, juvenile grasshoppers nearly double their body mass; however, since respiratory structures only increase during intermolt periods, oxygen delivery may be reduced. We show here that older grasshoppers have a reduced whole body air volume, yet femoral air volume is maintained through structural changes to the leg. In addition, older juveniles within an instar hop less frequently and produce more lactic acid during repeated jumping. We conclude that morphological changes during development may decrease convective oxygen delivery to the jumping muscle resulting in increased anaerobic metabolism and locomotory fatigue. These ontogenetic changes significantly alter the life‐history of juvenile grasshoppers. This research was partially supported by NSF IBN‐0104959 to SDK and JFH and the Union College IEF fund.
Bioenergetic profiles of isolated mitochondria from diapausing embryos of the annual killifish Austrofundulus limnaeus
Authors: Duerr Jeffrey M , Podrabsky Jason E
Published: June 2021

The annual killifish, Austrofundulus limnaeus, occupy ephemeral pond habitats by producing diapausing embryos which can reduce their oxygen consumption rate by up 90% compared to developing embryos. Mitochondria isolated from whole embryos collected at 10 dpf, diapause II, post‐diapause, diapause III, post‐hatch larvae, and adult liver exhibited oxygen consumption rates that increased with developmental stage in the presence of glutamate or succinate. Measurements of electron transport chain activity reflected life‐stage specific regulation. Minimum ETC activity was observed during diapause II: complex I activity was 12% of adult, complex II was 25% of adult, and complex IV activity was 20% of adult. Proton leak was significantly greater during diapause III and elevated in diapause II and larvae relative to 10 dpf and adult in the presence of glutamate, whereas no significant difference in life stage‐specific proton leak was observed in the presence of succinate. P:O ratios in the presence of glutamate or pyruvate and succinate were between 2 and 2.5 and between 1 and 1.2 respectively for diapause III embryos and adults. Overall, these data agree with whole‐embryo oxygen consumption rates previously measured and suggest a coordinated suppression of the mitochondrial electron transport chain and oxidative capacity during diapause. Supported by NSF grant IBN‐0344578 to JEP.
Adaptive Response with Oxidative Stress from CT Scans and Exercise in Mice
Authors: Phan Nghi , De Lisio Michael , Parise Gianni , Boreham Doug
Published: June 2021

We speculated that oxidative stress produced by repeated low dose diagnostic CT scans (10 mGy/scan, 2d/wk, 10 wk) would induce an adaptive response (AR) in C57BL/6 mice. We postulated that additional oxidative stress produced by exercise (1hr, 3d/wk, 10wk could further enhance the AR. We analyzed DNA double strand break (DSB) levels and genomic damage to stem cells via gH2AX foci and micronucleated reticulocytes (MN‐RET) flow cytometric assays, respectively. Following a challenge dose (1 Gy) in vitro, cells from CT mice had a significant increase (14%) in DNA DSBs compared to control and CT/exercised mice (p<0.035). A higher 2 Gy challenge dose showed that there was a significant reduction (16%) in DNA DSBs in CT/exercised mice (p = 0.002). When genomic damage was assessed, mice that had CT scans had significantly higher levels of MN‐RET (46% higher) compared to controls (p < 0.009). Interesting, the CT/exercise mice did not have any detectable increase in MN‐RET levels. After an in vivo 2 Gy challenge dose, control mice and CT mice had similar high levels of induced MN‐RET but there was a significant reduction (14%) in MN‐RET levels in the CT/exercise mice (p< 0.025). This research shows that CT scans can induce an AR in mice; however, this response is significantly modified by exercise.

This research was supported by US DOE, Low Dose Research Program (DE‐FG02‐07ER64343) and NSERC.
A conserved sequence in the argininosuccinate synthetase 3′ untranslated region binds specifically to carbamyl phosphate synthetase (ammonia), the first enzyme in the pathway of urea synthesis
Authors: Cohen Natalie S. , Zhang Zesong , Tran Kathy , Salles Arghavan
Published: June 2021

The five enzymes of urea synthesis in mammalian liver are in separate cellular compartments: the first two are in the mitochondrial matrix and the next three in the cytoplasm. The protein levels of all of the enzymes are coordinately regulated at both the transcriptional and post‐transcriptional levels. The pathway is highly ordered in situ, with the three cytoplasmic enzymes organized around the mitochondria. The mRNAs of two of the cytoplasmic enzymes, argininosuccinate synthetase (ASS) and argininosuccinate lyase, are also localized around the mitochondria. The ASS 3′ untranslated region (UTR) has a conserved sequence containing motifs implicated in post‐transcriptional regulation of various mRNAs in several organisms; we therefore did RNA/protein binding studies with this UTR and rat hepatocyte cytoplasmic extracts. Deletions and mutations showed that the conserved sequence was essential for protein binding. A 144 kDa protein that bound specifically to the ASS 3′UTR was obtained by RNA affinity chromatography, and identified by tandem mass spectrometry as an N‐terminally‐cleaved form of carbamyl phosphate synthetase (ammonia) (CPS I), the first enzyme of the pathway. Tissue transglutaminase bound the UTR non‐specifically. The results strongly suggest that there may be some CPS I in the cytoplasm that is involved in the post‐transcriptional regulation of ASS mRNA. Support: NSF #MCB 9601421 & 9983005.
Synthesis of the Ostreococcus tauri ADP‐glucose pyrophosphorylase genes and characterization of their expressed subunits
Authors: Kuhn Misty L , Falaschetti Christine A , Chaparro‐Krueger Cynthia , Ballicora Miguel A
Published: June 2021

The ADP‐glucose pyrophosphorylase small (S) and large (L) subunit genes from the unicellular algae Ostreococcus tauri were synthesized by PCR, subcloned, and the recombinant expression products characterized. This enzyme regulates the biosynthesis of starch in photosynthetic eukaryotes and has a reversible reaction in vitro. In higher plants, the L subunit makes the S subunit more sensitive to the activator. To understand the evolution of the subunit roles, we partially purified the S+L heterotetramer and S homotetramer from O. tauri, the simplest and smallest photosynthetic eukaryote. On the other hand, the L subunit could not be expressed alone with activity. We found that S+L is more sensitive to 3‐phosphoglycerate (activator) and phosphate (inhibitor) (A0.5 1.35 mM, I0.5 0.21 mM, forward direction, A0.5 0.80 mM, I0.5 0.51 mM reverse) than the homotetramer S (A0.5 2.20 mM, I0.5 1.40 mM forward, A0.5 2.80 mM, I0.5 1.20 mM reverse) with an activation of 33.5‐ and 7.6‐fold in the forward and 22.0‐ and 9.9‐fold in the reverse direction. The difference in regulation between the homotetramer (S) and heterotetramer (S+L) is not as pronounced as found in other plant ADP‐glucose pyrophosphorylases. These subunits in unicellular algae may not have evolved to play distinct roles as in higher plants. This research was supported by NSF grant no. MCB 0615982.
Intake of lutein and zeaxanthin differs with age, gender and ethnicity
Authors: Johnson Elizabeth J. , Maras Janice E. , Rasmussen Helen M. , Tucker Katherine L.
Published: June 2021

Lutein (L) and zeaxanthin (Z) are taken up into the macula where they may protect against age‐related macular degeneration (AMD).

OBJECTIVE: To determine intakes of L and Z, separately, in the National Health and Nutrition Examination Survey (NHANES) 2003–2004, using data from our laboratory on their levels in foods.

METHODS: The top 80 major food sources for L+Z intake in NHANES 2001–2002 were analyzed for L and Z by HPLC. Results were applied to dietary data from NHANES 2003–2004. Foods with L+Z were separated into L and Z in the nutrient database. Mean intakes from two non‐consecutive 24‐hr dietary recalls were grouped into food groups, based on nutrient composition; these were matched to the new database and L and Z intakes were calculated separately.

RESULTS: Among all age groups, both genders, and all ethnicities, L intakes were greater than Z. Relative intake of L to Z (L:Z) increased with age, with a L:Z ratio being greater in females. L:Z in Mexican Americans was 2‐ to 5‐fold lower than other ethnicities (other Hispanics, non Hispanic white, non Hispanic black, other).

CONCLUSIONS: Intakes of L and Z intakes differed among age groups, gender and ethnicities. Higher L:Z ratios were measured in groups at risk for AMD (older subjects, females). Estimates of intakes of L and Z are needed for determination of their individual role in eye health. (supported by USDA #581950‐7‐707, Chysantis, Inc)
The influence of mathematical ability and morning nutrition on mental arithmetic in preadolescents: An ERP study
Authors: Pivik R T
Published: June 2021

The effects of eating or skipping breakfast on ERP correlates of mental arithmetic were studied in preadolescents differing in experience (age) and mathematical skills. Participants, randomly assigned to treatment [eat (B) or skip (SB) breakfast (each, n = 41)], were sub‐grouped by age [8.8 yrs (B: n = 12; SB, n = 9), 9.7 yrs (B, n = 15; SB, n = 18), 10.5 yrs. (B, n = 14; SB, n = 14). At enrollment WRAT Arithmetic scores were superior in SB subjects. Addition and subtraction problems (randomized; 1–2 digit integers] followed by 3 possible answers were sequentially presented on a monitor (.2 sec; button press to indicate answer). Subjects were tested while fasting and again after treatment. Artifact‐free recordings (F3, F4, C3, C4, P3, P4) to correct‐answer subtraction problems were digitized and analyzed off‐line (ANOVA with post‐hoc t‐tests). Eating breakfast: a) attenuated or eliminated initial fasting between‐group ERP differences; b) affected component latencies in the youngest subjects [> early frontal (p ≤ .05), < late central (p = .02) relative to similar‐aged SB children]; and, c) improved performance in older children, i.e., increased number correct (9 yr group) and decreased reaction time (10 yr group). These findings suggest that morning nutrition facilitates mental arithmetic calculation and related response processes. Supported by USDA CRIS 6251‐51000‐002‐03S
The effects of a discretionary food allowance during a caloric restriction regimen with provided food
Authors: Gilhooly Cheryl Hajduk , Das Sai Krupa , Golden Julie K. , McCrory Megan A. , Dallal Gerard E. , Saltzman Edward , Kramer F. Matthew , Roberts Susan B.
Published: June 2021

The effects of self‐selected discretionary foods in a structured energy restricted diet on adherence to a caloric restriction (CR) regimen, dietary satisfaction, and weight loss were studied in 32 healthy, overweight (BMI 25–30 kg/m2) adults, aged 20–42y participating in the CALERIE trial. Subjects were randomized to either a high or low glycemic load diet at 70% of ad lib intake with food provided for 24 wks. During CR wks 15–20, subjects were randomized to receive 1000 kcal/wk of foods of their own choosing as part of their CR regimen (+C), or to receive no choice when the same amount of energy was provided as part of the meal plan (−C). Outcome measures included energy intake (EI), weight change, dietary satisfaction and eating behaviors. The +C group had significantly increased %EI from alcohol (p=0.04) and decreased %EI from protein (p=0.004) during the 6‐wk randomization compared to the −C group. No statistically significant differences were found between groups for changes in total EI, hunger, dietary satisfaction, eating behaviors or weight loss and there was no long‐term difference in weight change over time. These data suggest no detrimental effects of modest amounts of discretionary foods within a structured CR diet on weight loss, EI or dietary satisfaction. Due to high variability in the data and small sample size, further studies are needed to better understand how discretionary foods affect weight management.

Support NIH U01‐AG20480, the USDA 58‐1950‐4‐401
Spectral fingerprints for authentication and identification of variation in foods and botanical materials
Authors: Harnly James M , Luthria Devanand , Chen Pei , Lin Longze
Published: June 2021

Spectral fingerprints were acquired using ultraviolet/visible (UV/Vis), infra‐red (IR), near infra‐red (NIR), and mass spectrometry (MS) for broccoli, dry beans, and Ginkgo biloba with no prior chromatographic separation of the samples. Patterns were elucidated from the complex overlapping spectra using analysis of variance‐principal component analysis (ANOVA‐PCA). Using this approach, it was possible to detect differences in the chemical composition of the samples attributable to experimental factors such as the cultivar, growing conditions, growing site, plant‐to‐plant variability, and analytical variability. In addition, it was possible to determine the fraction of variance arising from each of the factors. All four analytical methods, including UV/Vis, were found to be equally effective for these analyses, although each focuses on different chemical components. An aqueous methanol extract was used for the UV/Vis and MS analyses, while finely powdered solid samples were used for IR and NIR. Only MS provided sufficient resolution to allow tentative identification of specific compounds.

This research was funded by USDA and the Office of Dietary Supplements at the National Institutes of Health.
A Fluorometric Assay to Determine Antioxidant Activity of Both Hydrophilic and Lipophilic Components in Plant Foods
Authors: Li Lei , Chen Chun Yen Oliver , Chun Hye‐Kyung , Cho Soo‐Muk , Park Ki‐Moon , Lee‐Kim Yang Cha , Blumberg Jeffrey B. , Russell Robert M. , Yeum Kyung‐Jin
Published: June 2021

We established a fluorometric method to determine total antioxidant activity of plant foods, green leafy vegetables and beans, after extracting [1] hydrophilic components (H) with acidified methanol (methanol:glacial acetatic acid:water = 50:3.7:46.3), [2] lipophilic components (L) with methanol followed by tetrahydrofuran (THF), or [3] both H + L using sequential extraction of acidified methanol and THF together. Both the hydrophilic assay (using a hydrophilic radical initiator, AAPH,10 mmol/L, and hydrophilic probe, DCFH, 2 μmol/L), and the lipophilic assay (using a lipophilic radical initiator, MeO‐AMVN, 2 mmol/L, and the lipophilic probe, BODIPY, 2 μmol/L) were used to measure antioxidant activity. The inhibition of BODIPY oxidation was significantly increased (p< 0.01) when both H + L were extracted using acidified methanol and organic solvent as compared to those extracted by organic solvent alone. In addition, the rate of DCFH oxidation was significantly delayed (p<0.05) when both components were present as compared to the H alone. The combination of H + L in these plant foods showed significantly greater antioxidant activity than that of either H or L alone. Thus, both H & L in plant foods and their interactions should be considered when determining antioxidant activity of plant foods.

[Supported in part by USDA #58‐1950‐7‐707 USA & BioGreen 21 Program (Code #20070301034009) RDA, Korea]
Chronic overload induced hypertrophy is associated with age‐related muscle mass loss and diminished mTOR signaling
Authors: Kendall Tracee Lynn , Brooks Naomi E , Fielding Roger A
Published: June 2021

The purpose of this study was to assess activation of the mTOR signaling pathway in young and aging rats in response to chronic muscle overload. Young (6 mo; n = 16) and older (30 mo; n = 23) male rats (F344×BN) were subjected to 4 weeks of bilateral surgical ablation (SA) of two‐thirds of the gastrocnemius muscle to promote compensatory hypertrophy in the plantaris (PLA) and soleus (SOL) or a sham surgery (CON). A significant interaction (p= 0.00) was found between group (SA vs. CON) and age (6mo. vs. 30mo.) when examining the normalized muscle weights (muscle weight/final body weight) of the PLA and SOL. The PLA muscle mass (mean ± SD) following SA was heavier in young rats when compared to aging rats (1.34 ± 0.07 g muscle weight/g body weight vs. 0.79 ± 0.08; p=0.00). An equivalent pattern was observed in the SOL in response to SA (0.52 ± 0.04 vs. 0.34 ± 0.04; p=0.00). Phosphorylation of p70S6Kin the SOL and PLA was similar in both young and aging rats when comparing SA (n=4) versus CON (n=4) (SOL: 6 mo; 0.21 ± 0.14 A.U. vs. 0.10 ± 0.10, 30mo; 0.20 ± 0.19 vs. 0.10 ± 0.06, PLA: 6 mo; 0.04 ± 0.02 vs. 0.02 ± 0.02, 30 mo; 0.06 ± 0.06 vs. 0.05 ± 0.01). These data indicate that aging muscle in rats is less responsive to hypertrophy in the PLA and SOL, which may be caused by impairment in mTOR signaling. Supported by USDA 58‐1950‐7‐707 and NIA AG‐25270
Transcriptional suppression of lipid‐induced apoptosis in skeletal muscle in vivo
Authors: Turpin Sarah Maggie , Ryall J G , Southgate R J , Darby I , Febbraio M A , Kemp B E , Lynch G S , Watt M J
Published: June 2021

Lipotoxicity occurs when fatty acid (FA) delivery exceeds the storage capacity of non‐adipose tissues, causing cell dysfunction and/or death by apoptosis. FA metabolites accumulate in skeletal muscle (SM) in obesity and cause lipid‐induced apoptosis in vitro.

AIM: To determine the lipotoxic effects of FAs in SM in vivo.

SM lipotoxicity was investigated in 3 models of FA oversupply: C57Bl/6 mice fed a high fat diet (HFD) for 12 weeks, genetically obese ob/ob mice, and acutely via 5 h Intralipid infusion in mice. Muscles were dissected then morphological, apoptotic and autophagic markers were assessed.

The Intralipid infusion increased plasma FAs 3‐fold and increased caspase 3 activity. HFD and genetic obesity induced adiposity and muscle triglyceride accumulation but did not induce apoptosis as assessed by TUNEL staining and caspase 3 activity. There was no evidence of SM regeneration or atrophy in HFD or ob/ob mice. The expression of 31 pro‐apoptotic genes were decreased by HFD. Autophagy was assessed by measuring LC3[alpha] and [beta] gene expression (conversion of [alpha] to [beta] represents autophagic activity). LC3[beta] was increased 3‐fold in ob/ob mice but there was no evidence of autophagy in HFD.

While lipid‐induced apoptosis occurs in vivo, these data suggest that SM adapts to chronic lipid oversupply by decreasing pro‐apoptotic gene expression to prevent cell death. Funded by: ARC, NHMRC
Modeling healthful eating: Links to parenting styles versus feeding practices differ as a function of child BMI
Authors: Danelia Ketevan , Hubbs‐Tait Laura , Moore Nicole Lee , Kennedy Tay S. , Baker Eric Ray , Larzelere Robert E.
Published: June 2021

This study compared predictors of parental modeling of healthful eating for preschool children with BMI < 50th versus BMI = 75th percentile, because this difference is linked to a six fold increase in later overweight. Participants were 131 3‐ to 4‐year‐old children with BMI measured in fall and spring. 53% had average BMI =75th percentile and 24% had average BMI < 50th percentile. Three parental styles ‐ authoritarian, authoritative, permissive ‐ and four feeding practices ‐ model, discourage, plan, external focus ‐‐ were measured. For children with BMI = 75th percentile, regression analyses showed parenting practices did not but feeding practices did predict modeling of healthful eating (R2 = .15, p=.016). For children with BMI < 50th percentile, regression analyses showed parental styles, particularly authoritative, predicted parental modeling of healthful eating behavior (R2 = .45, p =. 001) but feeding practices did not. Findings suggest nutrition educators working with families of heavier children could teach parental feeding practices to increase modeling. But, additional research should first investigate whether the authoritative practices of parents of normal weight children would enhance the healthful eating of heavier children. Funding: USDA‐CREES Number OKL0 2605.
The association between oxidative stress and pregnancy‐related symptoms of illness among vitamin A‐deficient women
Authors: Cheng Ting‐Yuan David , Schulze Kerry J , Arguello Margia A , Cui Hongjie , Khatry Subarna K , Christian Parul , West Keith P
Published: June 2021

Background

Vitamin A (VA) and beta‐carotene (BC) supplementation reduced pregnancy‐associated mortality and morbidity, as well as oxidative stress levels, in women of rural Nepal. This study examines the link between oxidative stress and symptoms of morbidity during pregnancy.

Methods

Serum malondialdehyde (MDA) was measured in a subset (n = 547) of women participating in a maternal VA or BC supplementation trial. Seven‐day frequencies of 21 symptoms were assessed within a month of blood collection. Total number of morbidity symptoms (0, 1, or =2), number of symptom‐days (0, 1‐9, or =10), and symptom clusters were modeled as dependent variables in multivariate logistic regressions.

Results

Serum MDA was lower among women with symptoms <20 weeks gestation, but higher among symptomatic women at =20 weeks. One square‐root unit increase in MDA (μmol/l) was associated with any symptoms (odds ratio [OR], 3.17; 95% CI, 1.04‐9.68) and upper gastrointestinal tract infection or nausea and vomiting (OR, 3.67; 95% CI, 1.07‐12.51) at =20 weeks of gestation. MDA was associated with number of symptoms (OR, 3.76; 95% CI, 1.45‐9.73) and symptom‐days (OR, 3.40; 95% CI, 1.36‐8.49). This trend held in three supplementation groups (VA, BC and placebo), but the effects were diminished in the VA group.

Conclusion

Oxidative stress was associated with morbidity symptoms. Supplementing VA/BC may mitigate the illnesses through reducing oxidative stress in this population. Funded by the Bill and Melinda Gates Foundation, Seattle, WA and the US Agency for International Development (USAID).
Differential Osteogenic Effect of Phytase Supplementation to a Low Zinc Diet in Rats
Authors: Antczak Amanda Jayne , Catrambone Daniel E , Loban Adrei , Hendrickson Nathan R , Young Andrew J , Scrimgeour Angus S
Published: June 2021

Adding vitamin D to a low calcium diet is known to effect trabecular but not cortical bone. Although adding phytase (P) to a low zinc (Zn) diet improves overall bone strength by increasing the bioavailability of Zn and other micronutrients, it is unknown if this supplementation has a differential effect on cortical and trabecular bone.

Purpose

Examine the effect of P‐supplementation to a low Zn diet in cortical and trabecular bone in both, L‐4 vertebrae and tibiae of rats.

Methods

Male Sprague‐Dawley rats were fed a diet containing either 5ppm Zn (ZnLo) (n=10), 5ppm Zn+1,500FTU/kg P (ZnLo+P) (n=10), or 30ppm Zn (ZnAd) (n=10) for 9 wks. In vivo peripheral quantitative computed tomography was used to scan right tibiae at 10% (trabecular parameters) and 50% (cortical parameters) of total. The L‐4 vertebrae were scanned at the 40, 50, and 60% of total length; an average of these represented the mid‐20% vertebral region. One way ANOVA determined group differences for cortical and trabecular bone mineral content (BMC), bone area (BA) and bone mineral density (BMD).

Results

Cortical parameters at the L‐4 were significantly higher in rats fed ZnLo+P compared to ZnLo diets. There was no significant difference between groups in trabecular parameters.

Conclusion

In contrast to the vitamin D:calcium relationship, P‐supplementation of a low Zn diet exhibited a positive effect on cortical BMD, BMC and BA at the L‐4 site, but had no effect on trabecular or tibial parameters.

The study was funded by the U.S.Army MRMC
Chinese Red Yeast Rice inhibits tumor growth and downregulates expression of genes for androgen and cholesterol biosynthesis in human prostate cancer xenografts in SCID mice
Authors: Hong Mee Young , Henning Susanne , Zhang Yanjun , Seeram Navindra P , Moro Aune , Heber David
Published: June 2021

Statin use has been associated with a reduced risk of prostate cancer. Chinese Red Yeast Rice (RYR) is a traditional food spice containing a family of eight monacolins one of which (monacolin K) is identical to lovastatin. The effects of 5% RYR in the diet on the growth of androgen‐dependent (LNCaP) and androgen‐independent (LNCaP‐AR) prostate cancer xenografts in SCID mice over 8 wks was examined. The expression of genes regulating androgen biosynthesis and cholesterogenesis were determined by quantitative real time PCR. RYR inhibited both androgen‐dependent and ‐independent prostate tumor xenograft growth by 54% and 40 %, respectively (P<0.05). RYR downregulated HSD3B2, AKR1C3 and SRD5A1 genes for androgen synthesis more than two fold in both tumor xenografts (p<0.05). RYR also downregulated HMGCR and SREBP‐2 genes involved in de novo cholesterogenesis (P<0.05). RYR downregulated androgen receptor gene expression only in androgen‐independent xenografts (P<0.05). Androgens are known to increase the growth of prostate cancer xenografts but this is the first study to demonstrate that RYR inhibits tumor growth and gene expression for both cholesterol biosynthesis and androgen biosynthesis. These studies demonstrate one of several possible pathways of RYR action in prostate cancer and suggest potential biomarkers for human studies. Supported by the UCLA CNRU CA 42710 and DOD W81XWH‐07‐1‐0158.
Localization of Myo 1 in Saccharomyces cerevisiae Mutants with a Multi‐budded Phenotype
Authors: Fields Whitney Cara , Zengel Janice , Lindahl Lasse
Published: June 2021

Evidence is beginning to accumulate suggesting that there is a connection between the cell cycle and ribosome biogenesis. The budding yeast Saccharomyces cerevisiae is an excellent model for understanding these highly conserved processes. During the yeast cell cycle, the newly forming cell appears as a bud on the mother cell and is released after mitosis and cytokinesis. It has been found that when the ribosomal protein L4 is depleted, an unusual multi‐budded phenotype is observed. It is not known exactly when during G1/S phase these multiple buds form. The source of this defect can be determined by observing the localization of different proteins in the cell after ribosome biogenesis has stopped. One such protein, Myo1, is known to be localized to the future bud site during the G1/S phase in the normally growing yeast cells. By tagging Myo1 with GFP and tracking its migration using confocal microscopy, it can be determined whether it appears at just one site or at multiple sites in cells depleted for L4. If it appears in multiple sites, then the defect must be occurring during the G1/S phase. These observations will be useful in understanding the signals between the cell cycle and ribosome biogenesis. This work was funded, in part, by NIH/NIGMS MARC U*STAR T34 08663 National Research Service Award to UMBC, NSF Grant MCB‐03449443 and the Howard Hughes Medical Institute.
Properties and Mechanism Studies of 5′‐Adenylylsulfate Reductases
Authors: Patel Raj B. , Kim Min K. , Kim Sang Gon , Kim Sung‐Kun
Published: June 2021

5′‐adenylylsulfate (APS) reductases is essential for cysteine biosynthesis in bacteria and plants catalyze the formation of sulfite from APS using either glutathione or thioredoxin. APS reductases from the bacteria Pseudomonas aeruginosa (PaAPR) and the green alga Enteromorpha intestinalis (EiAPR) and the archaeon Methanocaldococcus janishii were studied. EiAPR has a bipartite primary structure consisting of a domain with homology to PaAPR, and C‐terminal domain with homology to glutaredoxin. Iron and acid‐labile sulfide analysis indicated these enzymes both contain one [4Fe‐4S] cluster. Resonance Raman experiments confirmed the presence of a [4Fe‐4S] cluster. Redox titrations show that the EiAPR contains two disulfide/dithiol pairs with midpoint potential (Em) values of ‐290 mV and ‐140 mV, respectively. PaAPR contains only a single disulfide/dithiol cysteine pair with an Em value identical to that observed for the more negative component found in EiAPR. C‐terminal domain of EiAPR itself shows an Em value (i.e., ‐140 mV), consistent with the more positive EiAPR component, which is closer to that of E.coli glutaredoxin (i.e., ‐198 mV) than it is to E.coli thoredoxins (i.e., ‐270 mV). These results suggest that EiAPR has two different disulfide couples; one is like PaAPR and the other (i.e., the C‐domain site) may function in a manner similar to a glutaredoxin.

Supported by a grant from USDA and the Baylor University Undergraduate Research and Scholarly Activities Small Grant Program and the Vice Provost for Research.
Role of SsuF in the alkanesulfonate monooxygenase system from Pseudomonas putida
Authors: Millwood Mary Beth , Ellis Holly R.
Published: June 2021

The alkanesulfonate monooxygenase system from Pseudomonas putida enables the organism to utilize sulfonates and sulfate esters from the environment. The alkanesulfonate monooxygenase system is comprised of an FMN reductase (SsuE) and flavin‐dependent monooxygenase (SsuD). Although this system from Escherichia coli has been studied, the alkanesulfonate system from P. putida has not been evaluated. The system in P. putida possesses a third protein, SsuF, but the role of SsuF in the desulfonation mechanism has not been determined. SsuF has shown to have a high identity to a molybdopterin binding protein within E. coli. Previous studies have shown that SsuF is required for growth with sulfate esters and sulfonates, but the exact function of this protein has not been determined.

As P. putida possesses the additional SsuF protein, it is possible that the alkanesulfonate monooxygenase system may operate under an alternative mechanism than the E. coli system. Following the expression and purification of SsuF, several spectroscopic techniques were employed to analyze the structural and functional characteristics of the proteins. Further evaluation of SsuF was performed to determine the substrate range of SsuD and SsuE in the presence and absence of SsuF. Based on these studies, a functional role for SsuF in the desulfonation mechanism has been established.

Work supported by NSF Grant MCB545048.
p38 MAPK Regulates Sulfur Mustard‐Induced Cytokines Independent of NF‐κB, and Does Not Regulate Sulfur Mustard‐Induced Cell Death
Authors: Ruff Albert Leonard , Dillman James Franklin
Published: June 2021

Cutaneous and ocular injuries caused by sulfur mustard (SM, bis‐(2‐chloroethyl) sulfide) are characterized by severe inflammation and death of exposed cells. Given the known roles of p38 and NF‐κB in inflammatory cytokine production, and the known roles of NF‐κB and p53 in cell fate, these pathways are of particular interest in the study of SM injury. In this study, we utilized inhibitory RNA (RNAi) targeted against p38α, the p50 subunit of NF‐κB, or p53 to elucidate SM‐induced signaling mechanisms in normal human epidermal keratinocytes (NHEK). Analysis of culture supernatant from 200 µM SM‐exposed cells showed that inflammatory cytokine production was inhibited by p38α RNAi but not by NF‐κB p50 RNAi. These findings further support the critical role that p38 plays in SM‐induced inflammatory cytokine production in NHEK and suggest that NF‐κB does not play a role in the SM‐induced inflammatory response of this cell type. Inhibition of NF‐κB by p50 RNAi did, however, partially inhibit SM‐induced cell death, implying a role for NF‐κB in SM‐induced apoptosis or necrosis. Interestingly, inhibition of p53 by RNAi potentiated SM‐induced cell death, indicating that the role of p53 in SM injury is in cell survival and not cell death.

This research was supported by the Defense Threat Reduction Agency ‐ Joint Science and Technology Office, Medical S&T Division.
Culture duration and PUFA treatment influence expression of endocannabinoid proteins in MC3T3‐E1 osteoblast‐like cells
Authors: Hutchins Heather L , Zhao Dongjiao , Hannon Kevin M , Li Yong , Watkins Bruce A
Published: June 2021

Anadamide (AEA) and 2‐arachidonoylglycerol (2‐AG) are derived from arachidonic acid (AA). Recently, the endocannabinoid (EC) system has been linked to bone remodeling with in vitro and in vivo models. The purpose of this study was to examine the effects of culture time and n‐3 or n‐6 PUFA treatment on the EC system in the MC3T3‐E1 osteoblast‐like cell line. Cells (passage 7‐8) seeded at 6 x 104 in 6‐well plates were cultured in osteogenic medium up to 30 d, and at the last 72 h of incubation treated with AA, or eicosapentaenoic (EPA) plus docosahexaenoic (DHA) acids (10 µM each) with BSA as a vehicle control. Cells were harvested at 10, 25 and 30 d for fatty acid analysis and quantitative RT‐PCR. All cultures reflected enrichment of the respective fatty acid treatment; however, a greater magnitude of increase for EPA and DHA was achieved compared to AA. CB2 (the EC receptor) mRNA level was greatest in cells after 30 d compared to 10 d for the vehicle control; however, AA treatment decreased CB2 mRNA expression over time. Expression of the AEA synthesis enzyme, n‐acyl phosphatidylethanolamine phospholipase D (NAPE‐PLD), increased with AA treatment and culture time. These results show that developmental stage and PUFA alters the expression of CB2 and NAPE‐PLD, which suggests that maturation stage and PUFA treatments may influence the EC system in MC3T3‐E1 cells. Supported by USDA funds.
Water intake, nutrient intake, and body mass index among U.S. adults: Results from What We Eat in America/ NHANES 2005‐2006
Authors: Sebastian Rhonda S , Enns Cecilia Wilkinson , Goldman Joseph D.
Published: June 2021

This study sought to determine if intakes of water (moisture) from food and from beverages are associated with nutrient intakes and body mass index (BMI) among U.S. adults. Inverse associations between moisture from foods (but not beverages) and BMI have been detected by others, but those findings were not nationally representative for the U.S. Men and women age 19 years and older (n = 4,405) with one complete 24‐hour dietary recall in What We Eat in America (WWEIA)/NHANES 2005‐2006 were divided into quartiles based on their moisture intake from food and from beverages. Regression procedures adjusting for confounding factors were used to detect associations between moisture from each source and nutrient intakes and BMI. Nutrients studied were those for which >10% of Americans obtained less than the Estimated Average Requirement or Adequate Intake. Food moisture was positively associated with all nutrients analyzed (p < .01). Associations with beverage moisture were fewer and some were negative; e.g., higher beverage moisture was related to lower fiber intake in men. Beverage moisture was positively associated with BMI in men. No other associations between source of moisture and BMI were found. Overall, although higher one‐day food moisture intake is associated with higher intakes of selected nutrients, it is not associated with lower BMI in the U.S. adult population. Funding source for this study is USDA.
EPA and DHA status and indicators of atrophy on MRI scans of older adults
Authors: Arsenault Lisa Nicole , Scott Tammy M , Folstein Marshall F , Rosenberg Irwin H , Tucker Katherine L
Published: June 2021

Omega‐3 fatty acids may reduce the risk of Alzheimer's disease. Brain atrophy, particularly of the cortex and medial temporal lobe, indicates tissue pathology and is commonly seen on MRI scans of dementia patients. Our aim was to examine the cross‐sectional association between these fatty acids and MRI indicators of atrophy in 331 elderly home care clients in Boston, MA. Dietary intake was estimated from a validated, interviewer‐administered FFQ. Plasma phospholipid fatty acid profiles were assessed on a subset of subjects. Outcomes were based on clinical MRI measurements. After adjustment for major dementia risk factors, positive associations with total brain volume were seen for dietary EPA+DHA (β=0.84, P=0.02) and plasma phospholipid DHA (β=2.25, P=0.03). Higher sulcal grades, indicating more cortical atrophy, were negatively associated with dietary EPA+DHA (β=‐0.22, P=0.03) and plasma phospholipid DHA (β=‐0.56 (P=0.06). Hippocampal volume was positively associated with dietary EPA+DHA (β=0.007, P=0.01), but not with plasma fatty acids. EPA and DHA may play a role in preserving neuronal tissue, particularly the cortex, from damage and atrophy commonly seen in Alzheimer's disease. Longitudinal studies are needed to establish causality and confirm these associations.

This study was supported by NIA AG21790‐01, GCRC‐NIH MO1‐RR00054, and USDA‐ARS agreement 58‐1950‐7‐707.
Low energy reporting by adults in What We Eat In America, NHANES 2003‐2006
Authors: Rhodes Donna G , Nowverl Alvin B , Murayi Theophile , Moshfegh Alanna J
Published: June 2021

The U.S. Department of Agriculture Automated Multiple Pass Method (AMPM), a 5‐step method for collecting 24‐hour dietary recalls, was recently validated in a large doubly labeled water study and shown to be effective for collecting accurate group energy intake of adults. Since the AMPM is used to collect dietary intakes in What We Eat In America, National Health and Nutrition Examination Survey (NHANES), the objective of this research is to assess energy reporting accuracy in this nationally representative sample of the U.S. population. NHANES data were merged for the periods 2003‐2004 and 2005‐2006 and included men (n=4239) and non‐pregnant women (n=4084), 20 years and older. Energy reporting was assessed by computing a ratio of reported energy intake (EI) to estimated basal metabolic rate (BMR). EI was obtained from Day 1 dietary data and BMR was estimated using age and sex specific Schofield equations. The mean (95% CI) EI:BMR was 1.43 (1.41, 1.46) for men and 1.26 (1.24, 1.29) for women. Overall, for individuals with a body mass index (BMI) in the healthy weight category (18.5 to <25) the EI:BMR was 1.50 (1.47, 1.53). Using a Goldberg cutoff value of <0.9, 17% of men and 25% of women were classified as a low energy reporter (LER). For healthy weight individuals, less than 14% were LERs. The prevalence of low energy reporting was highest in women and in obese individuals (BMI=30). Other characteristics of LERs will be presented.

Funding source for this study is USDA
Modulation of gene expression by α‐tocopherol and α‐tocopheryl phosphate
Authors: Zingg Jean‐Marc , Meydani Mohsen , Azzi Angelo
Published: June 2021

The natural occurring vitamin E analogue, α‐tocopheryl phosphate (αTP), has been reported to be more potent in reducing cell proliferation and the expression of the CD36 scavenger receptor than the un‐phosphorylated α‐tocopherol (αT). We have now assessed the effects of αT and αTP on gene expression in THP‐1 monocytes using gene arrays covering essentially all human genes. More genes are regulated by αTP than by αT and no correlation was found between the expression levels of αT‐ and αTP‐regulated genes, suggesting that two different signal transduction pathways are affected. A group of genes was further confirmed by quantitative real time RT‐PCR. The expression of the vascular endothelial growth factor (VEGF) is induced by αTP at the mRNA and protein level. In vivo, the induction of VEGF by αTP may be relevant for cell survival and tissue homeostasis in several physiological conditions; strong induction of VEGF with higher concentrations of αTP could be used for therapeutic angiogenesis and show neuro‐, myo‐, and cardio‐protective effects. The mechanisms of VEGF regulation by αTP are currently further assessed. (This study is supported by USDA contract #58‐1950‐7‐707, and a fellowship to JMZ by Phosphagenics Ltd.).