bluuebunny/crossref_metadata_2025 huggingface dataset:
Compartmentalized glycogenolysis regulates lung cancer transcription
Authors: Young Lyndsay E.A. , Sun Ramon , Emanuelle Shane , Zhou Zhengqiu , Fillmore‐Brainson Christine , Gentry Matthew
Published: June 2021

The role of cellular metabolites in the direct control of epigenetic modifications is an emerging and rapidly evolving field. Herein, we identify a key function for nuclear glycogen in epigenetic regulation through compartmentalized acetyl‐CoA production and histone acetylation. In this study, we report glycogen accumulation in the nucleus of human non‐small cell lung cancer (NSCLC) specimens. To study this striking phenotype, we developed an innovative assay combining high‐purity sub‐cellular organelle isolation and stable isotope tracer technology to define the origin and metabolic fate of nuclear glycogen. We demonstrate that the nucleus is capable of carrying out glycogen synthesis, glycogenolysis and glycolysis, and both glycogen synthesis and glycolysis pathways require glucose‐6‐phosphate as the substrate. Nuclear glycogenolysis is dependent on ubiquitination and translocation of glycogen phosphorylase (GP) into the nucleus by malin, an E3 ubiquitin ligase. GP translocation is required for nuclear glycogen degradation and subsequent glycolysis to generate substrates for histone acetylation. Inhibition of nuclear glycogenolysis as a result of malin suppression is the cause of NSCLC nuclear glycogen accumulation. Re‐introduction of malin in model NSCLC cell lines restores nuclear glycogenolysis, results in increased histone acetylation, and delays cancer cell growth in vivo. This study uncovers a previously unknown foundational role for glycogen metabolism in the nucleus and elucidates another mechanism by which cellular metabolites control epigenetic regulation.

Support or Funding Information

The study was supported by the National Institute of Neurological Disorders and Stroke (grant ID: R01 NS070899‐06, P01 NS097197‐01), National Science Foundation (Grant ID: MCB‐1817414) and by the University of Kentucky Center for Cancer and Metabolism, National Institute of general medical sciences COBRE program (grant ID: P20 GM121327).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of aedeskinins and analogs on the basolateral membrane voltage (Vbl) of principal cells of Malpighian (renal) tubules of the adult yellow fever mosquito
Authors: Blum Austin William , Nachman Ronald J , Beyenbach Klaus W
Published: June 2021

Diuresis is stimulated in the yellow fever mosquito Aedes aegypti by the kinin peptides. All three aedeskinins depolarize the transepithelial voltage (Vt) in Aedes Malpighian tubules (Veenstra et al., 1997), like leucokinin (Pannabecker et al., 1993). The effects of aedeskinin II and III, a synthetic analog 1460II, as well as leucokinin‐VIII (kinin of cockroach) on the basolateral membrane voltage (Vbl) of principal cells of Malpighian tubules of Aedes aegypti were studied. Both aedeskinins and synthetic analog 1460II hyperpolarized Vbl, similar to the effect of leucokinin‐VIII (expected from depolarization of Vt). Table 1 lists the peptides used in the present study.

The kinin receptor agonist 1460 II was synthesized through the replacement of á amino acids with β amino to resist digestion by proteases. The substitution does not abolish voltage activity of the peptide, even though the substitution is within the conserved pentapeptide sequence F‐X‐X‐W‐Xa needed for diuretic action. The reversibility of the effects of 1460II indicates that the substitution does not bind the peptide irreversibly to the receptor. Supported by NSF Grant IOB‐0542797 to KWB.
Extreme blood oxygen depletion in diving elephant seals
Authors: Meir Jessica Ulrika , Champagne Cory D. , Costa Daniel P. , Ponganis Paul J.
Published: June 2021

The repetitive, long dives of elephant seals, Mirounga angustirostris, have remained a physiological enigma in that oxygen (O2) stores appear inadequate to maintain aerobic metabolism. To investigate blood O2 depletion in this species we 1) measured venous O2 partial pressure (PO2) continuously during dives with a backpack PO2/temperature recorder and, 2) characterized the O2‐hemoglobin (Hb) dissociation curve. Similar to other marine mammals, O2‐Hb dissociation curve results were: P50 (50% saturation) ~ 31 mmHg, Bohr effect (P50) ~ −0.6. End‐of‐dive PO2 routinely reached 2–10 mmHg in the extradural vein and hepatic sinus, demonstrating remarkable hypoxemic tolerance in this species. Based on prior studies, we hypothesize that arterial PO2 also reaches similarly low levels. These data allow calculation of % Hb‐saturation, the magnitude of blood O2 store depletion during dives, and the contribution to diving metabolic rate. Venous temperature during routine dives remained near 37°C, suggesting that hypothermia‐induced hypometabolism is an unlikely contributor to metabolic suppression while diving. We believe the elephant seal's extreme hypoxemic tolerance allows near complete depletion of its previously measured large blood O2 store, providing sufficient O2 to maintain metabolism during repetitive, long dives. Funding: NSF grant IOS‐0641801 & SIO Halliday Student Field Research Award
Understanding alternate policy approaches for improving the school food environment
Authors: Shroff Monal , Jones Sonya J , Frongillo Edward A
Published: June 2021

Multiple calls for action emphasize the need for policy solutions to regulate the school food environment in the United States, especially the sales of competitive foods. Policy evaluation is an important step towards understanding and developing a strong policy solution including finding effective policy approaches. This study examines bills introduced through state legislation and characterizes the bill content for various policy approaches/designs. Based on Howlett's (2000) conceptualization, bill content was characterized to possess "substantive" and "procedural" features and also coded for the kind of governing resources (information, authority, treasure, or organizational resource) used for the effectiveness of the approach. We coded 43 bills passed between 2001 and 2003 from 50 states which addressed the regulations of competitive foods in schools. Most competitive food legislation has multiple substantive and process features. Authority (e.g., forming a task force) is the main governing resource in bills with procedural features, while monetary compensation or training is mainly used as a resource in bills with a substantive features. Results of the study will provide better understanding of how policy should be used to regulate the school food environment and the impact of such policy approaches on child health outcomes. Funded by USDA/ERS.
Modulation of the blood‐brain barrier (BBB) charge by plasma glycoprotein orosomucoid
Authors: Fu Bingmei , Yuan Wei , Li Guanglei
Published: June 2021

To investigate how orosomucoid modulates the charge of the cerebral microvessel wall or the BBB, we measured the apparent permeability (P) of pial microvessels to various charged molecules: FITC‐α‐lactalbumin (MW14k, net charge = ‐11), FITC‐ribonuclease (MW13.7k, +3), FITC‐albumin (MW65k, ‐19) in the perfusate of either 1% bovine serum albumin (BSA) only or that additionally containing 0.1mg/ml orosomucoid. All in vivo experiments were performed on SD rats (250‐300g). The pial circulation was observed through a section of thinned frontoparietal bone while the fluorescently labeled solute was injected through the carotid artery (Yuan et al., 2008). P was determined using a quantitative fluorescence imaging and analyzing method. In the absence of orosomucoid, the mean P to positively charged ribonuclease was four times as large as that to negatively charged α‐lactalbumin although they have similar size, while in the presence of orosomucoid, P to ribonuclease increased to almost nine times of that to α‐lactalbumin. However, P to negatively charged albumin in the presence of orosomucoid was only half of that in its absence. Our results suggest that orosomucoid increases the negative charge of the BBB, which is consistent with that reported in Curry et al. (1989) for mesenteric microvessels. Supported by NSF and Andrew Grove Foundation.
Diurnal rhythms of muscarinic receptor expression in the eye of zebrafish
Authors: Capalbo Elizabeth Louise , Garcia Dana M
Published: June 2021

In teleosts, pigment granules disperse into the apical projections of the RPE to protect photoreceptors from photobleaching in ambient daylight. Activation of muscarinic receptors induces light‐adaptive pigment granule dispersion in isolated RPE. In order to determine whether muscarinic receptors could be involved in acetylcholine‐mediated pigment granule dispersion in vivo, we looked at muscarinic receptor expression in zebrafish eyes at time points in which the pigment granules of the RPE are fully dispersed or fully aggregated, midday and midnight, respectively. We used qRT‐PCR to study the differential expression of muscarinic receptors. Our data indicate that M3 muscarinic receptor RNA is expressed at higher levels at midnight than midday. Studies are underway to determine whether other subtypes of muscarinic receptors are also differentially expressed and to localize expression of the receptors in retinal sections using in situ hybridization and immunofluorescence microscopy. This work was supported by NSF IOB 0615762.
Regulation by unsaturated fatty acids of the expression of key genes involved in HDL metabolism
Authors: Kuang Yu‐Lin , Lichtenstein Alice H. , Lamon‐Fava Stefania
Published: June 2021

In humans and animal models, polyunsaturated fatty acids (PUFA) have been suggested to influence the expression and/or activities of proteins involved in the metabolism of high‐density lipoprotein (HDL). The current study determined the effects and the mechanisms of action of different PUFA on genes regulating HDL metabolism in human hepatoma cells (HepG2). HepG2 cells cultured for 24 hr in 200 uM docosahexaenoic acid (DHA), eicosapentaenoic acid or arachidonic acid (AA) had lower mRNA levels of apolipoprotein A‐I (apo A‐I) (30‐40 %, P = .01), ATP‐binding cassette A1 (ABCA1) (35‐50%, P = .05), lecithin: cholesterol acyltransferase (LCAT) (40‐50%, P = .001), and phospholipid transfer protein (PLTP) (25‐40%, P = .05) compared to those exposed to palmitic acid, as determined by real‐time PCR. In transfection assays, 200uM AA or DHA promoted peroxisome proliferator activated receptor (PPAR) and suppressed liver X receptor activities. HepG2 cells transfected with plasmids containing serial deletions of the apo A‐I promoter showed that the element mediating the PUFA‐related transcriptional repression is located in the ‐192 to ‐41 region of the promoter. In conclusion, very long‐chain n‐3 and n‐6 PUFA slowed the formation and the maturation of HDL particles. These FA also downregulated apo A‐I mRNA level and promoter activity via mechanisms other than PPAR activation. Funded by USDA and Unilever Health Institute.

Grant Funding Source

USDA
Effect of vitamin E and SNPs at cytokine genes on cytokine production in the elderly
Authors: Belisle Sarah E. , Leka Lynette S. , Delgado‐Lista Javier , Jacques Paul F. , Ordovas Jose M. , Meydani Simin Nikbin
Published: June 2021

Vitamin E (E) impacts cytokine production, yet individual response to E supplementation varies. Studies show cytokine production is heritable. Common single nucleotide polymorphisms (SNPs) may explain differences in cytokine production between individuals. We hypothesize that differential response to immunomodulatory actions of E reflect genetic differences among individuals, including SNPs at cytokine genes, which modulate cytokine levels. We used data from an E intervention in the elderly to test our hypothesis. We observed the effect of E on TNF‐α production depended on TNF‐α ‐308G>A. Subjects with G/G genotype at TNF‐α ‐308G>A who were supplemented with E had higher TNF‐α compared to subjects with G/G genotype given placebo. In contrast, subjects with A/A or A/G genotype at TNF‐α ‐308G>A supplemented with E had lower TNF‐α compared to subjects with A allele given placebo. This suggests individual immune response to E is in part mediated by genetics. Since A allele at TNF‐α ‐308G>A has been associated with higher TNF‐α, our results may indicate that anti‐inflammatory effect of E is specific to those genetically predisposed to higher inflammation. Studies on the mechanism driving the E and TNF‐α ‐308G>A interaction, and its disease implications are needed. Support: NIA NIH grant 5R01‐AG013975; USDA agreement 58‐1950‐7‐707; a DSM Nutritional Products, Inc. scholarship. CIBEROBN is an initiative of ISCIII.
Household food security is associated with early childhood language development in rural Bangladesh
Authors: Saha Kuntal K. , Tofail Fahmida , Frongillo Edward A. , Mehrin Fardina , Arifeen Shams E. , Persson Lars Å. , Rasmussen Kathleen M. , Hamadani Jena D.
Published: June 2021

Although household food security (HHFS) has been linked to academic performance in school children, its association with early childhood development has received less attention. We studied this association in 1439 infants born in 2002‐2003 in the Maternal and Infant Nutrition Intervention in Matlab study in Bangladesh. A HHFS scale was created from data on HHFS collected from their mothers during pregnancy. Infants' cognitive and motor development was assessed at 7 mo using a means‐end‐problem‐solving test and Bayley Scales of Infant Development‐II (BSID‐II), respectively, and was reassessed at 18 mo using BSID‐II and a language test based on MacArthur's Communicative Development Inventory. We used linear regression, adjusting for potential confounders. HHFS was positively associated with language expression (P < 0.01) and comprehension (P < 0.05) at 18 mo, but not with mental or motor development at 7 mo or 18 mo of age. These findings emphasize the importance of HHFS during pregnancy for early childhood development and also provide a strong reason to ensure HHFS in low‐income countries for optimum childhood development. (MINIMat was funded by ICDDR,B, UNICEF, Sida‐SAREC, UK MRC, Swedish Research Council, DFID, CHNRI, Uppsala Univ. & USAID.)
The influence of extein residues on the protein splicing of three Pyrococcus abyssi inteins
Authors: Kerrigan Adam M. , Powers Taryn L. , Dorval Deirdre M. , Reitter Julie N. , Mills Kenneth V.
Published: June 2021

Protein splicing is catalyzed by an intervening polypeptide, called an intein. Inteins facilitate their own excision from the flanking polypeptides, called exteins, as well as the ligation of the exteins. Three inteins interrupt the ribonucleotide reductase of the extreme thermophile Pyrococcus abyssi. We have separately over‐expressed each intein in E. coli as a fusion protein with flanking affinity domains. By site directed mutagenesis, we have altered the sequences flanking each intein to determine the flexibility of the intein insertion site. We have also made mutations to residues suspected of catalyzing the splicing steps in each intein. We plan to study the influence of these mutations on the rate and extent of splicing. This material is based upon work supported by the National Science Foundation under Grant No. 0320824 and CAREER grant No. 0447647.
Whole grain consumption in adults: Results from NHANES 1999‐2004
Authors: ZANOVEC MICHAEL , O'Neil Carol E. , Cho Susan E. , Nicklas Theresa A.
Published: June 2021

The objective of this study was to examine the effect that whole grain (WG) consumption has on diet quality and nutrient intake in a recent, nationally representative sample of adults: 19 to 51 years (n=7039) and 51+ years (n=6,237) using a secondary analysis of NHANES 1999‐2004 data. Participants were divided into four WG consumption groups: >0 to <0.6, >0.6 to <1.5, >1.5 to <3.0, and >3.0 servings/day. Nutrient intakes and diet quality, using the Healthy Eating Index, were determined for each group. Sample‐weighted percentages in WG consumption groups were calculated using energy, ethnicity, gender, and age as covariates. Least‐square means were calculated. P for linear trend analysis was determined using WG intake as a linear covariate. Adults 19‐50 and 51+ years consumed an average of 0.63 and 0.77 servings of WG per day, respectively. For both age groups, diet quality, energy, fiber, and polyunsaturated fatty acids increased significantly with an increased number of servings of WG consumed. Intake of total sugars (19‐50 years only), added sugars, saturated fatty acids, monounsaturated fatty acids, and cholesterol decreased with an increased number of servings of WG consumed. Intake of all micronutrients, except vitamin B‐12 and sodium, increased significantly with consumption of increasing servings of WG. Supported by USDA and Kellogg's Corporate Citizenship Fund.
The onset and magnitude of cardiovascular drift depend on exercise intensity in competitive cyclists.
Authors: Murphy Kristi , Winger Jena , Stavrianeas Stasinos
Published: June 2021

PURPOSE

To examine the potential effects of cardiovascular drift (CVD) on aerobic and anaerobic metabolism.

METHOD

Competitive cyclists (n=11, age 40.3±10.9years, height 176±10cm, weight 70.9±11.6kg, VO2max 56.31±8.48ml▸kg−1▸min−1) performed 5 tests on a cycle ergometer 3‐5 days apart: a VO2max test (2‐min stages to exhaustion), a Maximal Lactate Steady State (MLSS), and 3 prolonged (90 min) submaximal bouts at 95%, 100%, and 105% of the MLSS (average: 190W, 199W, and 209W). HR, and VO2 were recorded continuously during the first 2 tests, the first 35 min and every 10 min thereafter for the 3 longer bouts. Blood lactate was measured at the end of each stage for the first 2 tests, and every 10 min for the 3 longer bouts.

RESULTS

Only 3 cyclists completed the 105% MLSS bout, so comparisons were only made over 35 min. Onset of CVD was different for every participant at each intensity. Small changes in load (<10W) yielded differences (p < 0.05) across the 3 intensities in CVD amplitude (6 BPM, 8 BPM, and 9 BPM for the 95%, 100%, and 105% MLSS), VO2 (no change, 2 ml▸kg−1▸min−1, and 4 ml▸kg−1▸min−1), and blood lactate (0.3 mmol▸L−1, 0.5 mmol▸*;L−1, and 0.9 mmol▸L−1) between plateau and minute 30.

CONCLUSION

We successfully measured the small changes in the metabolic cost during CVD. The onset of CVD is affected by the intensity of exercise and the amplitude of CVD is greater as exercise intensity increases.

Supported by the Mary Stuart Rogers Foundation and an NSF grant (DUE 0511219).
Protein misfolding and aggregation: Insight from single molecule study
Authors: Lyubchenko Yuri L , Yu Junping , Kim Bo‐Hyun
Published: June 2021

Protein misfolding is the very fist step in a long chain of the aggregation events eventually leading to neurodegeneration. The structure of individual protein molecules within well ordered aggregates can be partially elucidated using established techniques, however, we still lack of a mechanistic understanding of the process leading to the misfolded conformations of a protein and their role in the aggregation. Our central hypothesis is that proteins in misfolded states are characterized by elevated interprotein interactions that can be detected by AFM force spectroscopy approach. We developed a novel nanoprobing AFM approach capable of detecting and analyzing transient misfolded states of the proteins differed from the normal ones by increased interprotein interactions. The proteins were immobilized covalently at the N or C termini at the AFM probes and substrates and their interactions were measured by multiple approach‐retractions cycles performed by the AFM instrument. These experiments applied to α‐synuclein and amyloid β‐peptide revealed enormously high stability of misfolded dimers suggesting their critical role in the early stages of the aggregates formation. A high stability of the misfolded dimers is unexpected finding that sheds a new light into the mechanisms of the self‐assembly of misfolded proteins into the disease related aggregates. Supported by DOE grant. DE‐FG02‐08ER64579.
Characterization of metal‐reducing enzymes expressed by the bacterium Shewanella algae: Gene sequencing
Authors: Sohn Changmo , Tesch Sarah K. , Penny Jonathan T. , Goyne Thomas E.
Published: June 2021

This work is a small part of a large Department of Energy (DOE) research effort to harness microbes to the task of immobilizing actinide contaminants (chiefly uranium and plutonium) in soil and ground water. DOE's long term goal is to use microbe‐driven actinide immobilization to help contain the enormous subsurface pollution problem that resulted from Cold War‐era nuclear weapons fabrication and testing. The envisioned immobilization mechanism depends on microbes using water‐soluble actinide species in high oxidation states (for example, U+6) as terminal electron acceptors to yield less soluble species (for example, U+4). The specific goal of the work reported here is to sequence the genes that code for metal‐reducing enzymes expressed by the Gram‐negative soil bacterium Shewanella algae. Using a PCR‐based approach, we have so far partially sequenced two genes that are highly homologous to metal‐reducing genes in S. algae's closest relatives (specifically, homologs of the MtrA and MtrB genes in the S. onidensis genome). As soon as we have the complete gene sequences in hand, we intend to use a combination of SDS‐PAGE and mass spectroscopy to study the expression of these proteins. This work is supported by DOE subcontract 21059‐001‐05.
Added sugars and solid fat replace fruits, vegetables, and milk in the diets of preschool‐age children, especially that of low‐income children
Authors: Bowman Shanthy , Friday James , Martin Carrie
Published: June 2021

The study objective is to examine the associations between discretionary calories and fruit, vegetable, and milk intakes in children, ages 2 to 5 years, from low‐ (<131% of poverty; n=394), medium‐ (131‐350%; n=233), and high‐ (> 355%; n=107) income households. What We Eat In America, National Health and Nutrition Examination Survey, 2003‐2004 data were used. Regression models adjusting for age and gender were used, with alpha =0.05 level of significance. Low‐ and medium‐income children had significantly more energy from added sugars (265±12, 252±18, and 203± 9 kcal) and solid fat (388±15, 383±32, 305±17 kcal) than high‐income children, respectively. Low‐income children had the highest total energy intake (1804± 47, 1689± 56, 1572± 59 kcal). For every 10% increase in percent total energy from added sugars, fruit intakes reduced by 0.9, 0.5 and 0.6 cup and milk intakes reduced by 0.4, 0.6 and 0.5 cup, for low‐ to high‐income children, respectively. For every 10% increase in percent total energy from solid fat, fruit intakes reduced by 1.2, 0.7, and 0.5 cup, and milk intake increased by 0.5 and 0.2 cup in low‐ and medium‐income children. The reduction in vegetable intakes of low‐ and medium‐income children was small, but significant. Food habits form early in life. The study showed the need for preschool‐age children, especially those form low‐income households to reduce added sugars and fat intakes. Study funded by USDA.
Differential effects of chronic overload‐induced muscle hypertrophy on mTOR and MAPK signaling pathways in adult and aged rats
Authors: Chalé‐Rush Angela , Morris Evan P , Kendall Tracee L , Brooks Naomi E , Fielding Roger A
Published: June 2021

We examined activation of the mammalian target of rapamycin (mTOR) and mitogen‐activated protein kinase (MAPK) signaling pathways in adult (Y; 6 mo old; n = 16) and aged (O; 30 mo old; n = 16) male rats (Fischer 344 x Brown Norway) subjected to chronic overload‐induced muscle hypertrophy of the plantaris (PLA) and soleus (SOL) muscles. Animals underwent either 4 weeks of bilateral surgical ablation (SA) of two‐thirds of the gastrocnemius muscle or sham surgery (CON). Animals were subjected to SA to promote compensatory hypertrophy in the PLA and SOL muscles. Muscle weights of the Y CON (434 ± 39 mg; mean ± SD; p < 0.05) and Y SA (506 ± 56 mg) rats were significantly heavier than the O CON (353 ± 18 mg) and O SA (402 ± 41 mg) rats. There was a significant interaction between group and age for phosphorylated mTOR. Phosphorylated mTOR was higher in the Y SA group (4129 ± 674 A.U.; mean ± SE; p < 0.05) than in the Y CON (1582 ± 674), O SA (2063 ± 630) and O CON (2667 ± 728) groups. The ratio of phosphorylated to total mTOR was significantly higher in both SA groups than in their respective controls (0.107 ± 0.105 A.U. vs. ‐0.297 ± 0.112; p < 0.05). We also observed a significant effect of age on total p44 MAPK. It was higher in the O (17969 ± 1242; p < 0.05) than the Y (13342 ± 1242) rats. These data indicate mTOR signaling is attenuated in an aging animal model of chronic overload‐induced muscle hypertrophy. Supported by USDA 58‐1950‐7‐707, NIA AG‐25270 and DK007651.
Ontogeny of the desaturase‐elongase pathway in neonatal pigs fed arachidonic acid
Authors: Jacobi Sheila K. , Lin Xi , Corl Benjamin , Odle Jack
Published: June 2021

This study evaluated the effects of supplementation of ARA on delta‐6‐desaturase (D6D) and delta‐5‐desaturase (D5D) mRNA abundance and synthesis of ARA in the intestine and liver. Day old pigs (n=96) were fed a milk based formula for 4, 8, and 16 d. Diets contained either no polyunsaturated fatty acids (0% ARA, negative control), 0.5% ARA, 2.5% ARA, 5% ARA or 5% eicosapentaenoic acid (EPA) of total fatty acids. A sow fed reference group (SRC n=18) also was included. The mRNA relative abundance increased differentially (diet x age interaction, P < 0.05) from d4 to d16 in pigs fed 0.5% (D6D‐55%, D5D‐66%) versus 5% (D6D‐60%, D5D‐64%) ARA diets in the intestine. Transcript abundance of D6D and D5D were not affected by diet in the liver (P>0.05), however, there was a precipitous decrease between birth and d4 (~5‐fold, P<0.05). To measure flux through the desaturase‐elongase pathway we incubated liver and intestinal mucosa with 13C‐linoleate(C18:2, n‐6) and traced its metabolism to ARA via GC/MS. In the intestine accumulation rate (nmol/g tissue/h) of 13C‐ARA was not affected by piglet age, but doubled in pigs fed 5%ARA compared with pigs fed 0.5% ARA. Furthermore, accumulation of 13C18:3, n‐6 indicated that, among ARA fed pigs, D6D activity was highest in those fed 5% ARA, and this activity was equal to that measured in SRC pigs (P<0.05). Supported by CSREES, USDA NRI 2005‐35200‐16174.
Knowledge and attitudes about traditional African vegetables are related to their consumption in Kenya and Tanzania
Authors: Herforth Anna , Low Jan , Kaguongo Wachira , Pletziger Stefan , Njenga Mary
Published: June 2021

This study compares the frequency of consumption of traditional African vegetables (TAVs) in Kiambu, Kenya, and Arusha, Tanzania, as it relates to attitudes about the status of these vegetables, and knowledge about their nutritional value. The study is done within the context of a program promoting TAVs (Amaranthus spp., Solanum spp., Cleome gynandra, Vigna unguiculata, Ipomoea batatas) based on their nutritional attributes, including high micronutrient content and potential to improve overall diet quality. A baseline survey was administered in 2007 to 360 small farmer households, some of whom had been previously exposed to the promotion of traditional vegetables. The survey, as well as focus group discussions, gathered information on knowledge and attitudes about the vegetables, and diet. Survey data show that greater nutritional knowledge and more favorable attitudes about the TAVs are highly significantly predictive of TAV consumption at the household level. Qualitative data support these associations, and indicate that knowledge and attitude changes precede consumption changes. These findings suggest that promotion of crops based on nutrition and image is likely to be successful in increasing consumption of those crops among small farmer households in the study regions.

Funding sources: Kilimo Trust; NSF; HE Babcock Chair and Einaudi Center, Cornell University
DNA damage determined by Single cell gel electrophoresis using cryopreserved lymphocyte, frozen whole blood in EDTA tube or DNA stabilized tube
Authors: Ronchi Carlos Fernando , Berchieri‐Ronchi Carolina Bragiola , Correa Camila Renata , Ferreira Ana Lucia dos Anjos , Zhao Yan , Kim Sung Woo , Yeum Kyung‐Jin
Published: June 2021

Oxidative DNA damage is of great importance due to the growing recognition that such damage can initiate and promote chronic diseases. To determine the usefulness of frozen whole blood on the analysis of DNA damage, lymphocytes isolated from the fresh blood followed by cryopreservation as well as collected from the frozen whole blood were analyzed in parallel for DNA damage. Lymphocytes were isolated from fresh blood from sows (n=10) and cryopreserved (Cryo). Whole blood samples were also collected in EDTA‐tubes (EDTA) as well as DNA‐stabilized tubes (DNA). Cryopreserved lymphocytes as well as lymphocytes recovered from the frozen whole blood were analyzed by alkaline single cell gel electrophoresis (comet assay) to determine the single and double strand brakes. The comet assay indicated less DNA damage (% damage) in the lymphocytes obtained from the frozen whole blood in DNA‐tube as compare to those of cryopreserved lymphocyte (Cryo, 45.8 ± 5.88 > DNA, 37.52 ± 6.72 = EDTA, 40.1 ± 3.65, p<0.05). Similar results obtained when the DNA was challenged with H2O2 (Cyro, 51.9 ± 2.04 > DNA, 43.3 ± 5.43 = EDTA, 40.7 ± 6.42, p<0.05). The current study indicates that frozen whole blood can be used for DNA damage analysis and DNA‐preservation tube shows better response to the oxidative stress challenge. The presence of antioxidants in the whole blood may have a protective effect against oxidative DNA damage that might occur during the storage, which warrants further study. [Supported by CAPES0635‐09, FAPESP2008/08199‐2, USDA 1950‐51000‐065‐08S]
Identification of bioactive dietary and natural components for targeting cancer stem cells
Authors: Zhou Jin‐Rong , Li Yanli , Gong Yi , Blackburn George
Published: June 2021

Cancer stem cell (CSC) is a small specific subset of cancer cells and is suggested to be responsible for initiation, progression, drug resistance and recurrence of cancer. The search for novel agents to inhibit CSC self‐renewal has emerged as a research priority in cancer prevention and treatment. The objective of this study was to identify active dietary and natural compounds against CSC. Human breast, prostate, pancreatic and lung cancer cell lines were cultured at non‐adherent, self‐renewal conditions. The tumorsphere forming efficiency was determined to evaluate the effect of dietary and natural compounds on CSC self‐renewal. The expression of CSC‐related genes was determined by quantitative real time‐PCR. Tumorsphere formation was associated with altered expression of CSC‐related genes. Dietary components genistein and myricetin showed limited activities inhibiting CSC self‐renewal; sulforaphane and EGCG showed moderate activities. Tanshinones from Salvia Miltiorrhiza (tanshinone I, tanshinone IIA and cryptotanshinone) showed potent activities in inhibiting self‐renewal of CSCs in different cancer cell lines, with tanshinone I being the most potent. The inhibition of CSC self‐renewal was associated with modulation of gene expression of certain CSC‐related biomarkers. Our results warrant further evaluation of tanshinones as candidate anti‐CSC agents. Supported in part by DOD (PC073988).
Polonium‐210 in Diesel Particulate Matter (DPM)
Authors: Lykken Glenn I. , Czapiewski Robert , Ward Thomas
Published: June 2021

Inhalation of DPM has been reported to shorten the life of 70,000 Americans each year and to increase the risk of developing brain tumors in children. Environmental radon progeny (ERnP) have been measured in human brain tissues, but how they entered the brain remains uncertain. We hypothesize ERnP may be incorporated in DPM during combustion and enter the body through inhalation. Therefore, we captured DPM on 30 nm filters. The DPM, containing polonium‐210 (Po‐210) came from a variety of diesel engines operating in normal outdoor atmospheres with radon concentrations from 7 to 33 Bq/m3. Initially, the Po‐210 alpha emissions were measured directly from the nanofilters. However, the activity dropped dramatically, presumably due to the volatility of the DPM particles. Therefore, immediately after loading the nanofilters were soaked for approximately 12 hours in nitric acid. Po‐210 from this solution was deposited onto silver disks and counted. The observed Po‐210 activities were dependent upon the type of diesel engine and the atmospheric ERn concentration. The work was sponsored in part by the USDA ARS GFHNRC.
Interactions among FliN, FliH and FliI of the Salmonella enterica flagellar export apparatus govern secretion
Authors: Wilson Jo Leanna , Scott Israel M. , McMurry Jonathan L.
Published: June 2021

Using two optical biosensing methods, surface plasmon resonance and biolayer interferometry, a study of the interactions among proteins involved in flagellar type III secretion was undertaken to understand the mechanism of substrate delivery to the membrane‐embedded export complex. An interaction between C‐ring protein FliN and export apparatus protein FliH has been proposed to be a mode of targeting substrates via a complex that is necessarily dynamic as the position of FliN at the base of the C‐ring is too far from the membrane to allow the complex to remain associated during export. Both techniques allow quantitative measurement of association and dissociation. Rate and affinity constants are reported for pairwise interactions; both FliN‐FliH and FliH‐FliI have submicromolar KDs. Experiments investigating the conditions necessary to dissociate FliN from FliH or FliH from FliI are described. Substrates were screened for binding to FliH and FliI in the presence and absence of FliN. Kinetic analysis will allow us to rank affinities, determine how binding changes in the presence of other participants, search for export signals within substrates etc., developing a better understanding of type III secretion. This work was supported by grants from the Public Health Service (R15GM080701), NSF (CHE 0922699), the Research Corporation (CC6900) and a KSU Mentor‐Protégé Grant (to J.L.W.).
Determining retinol and 3‐hydroxy‐retinol concentrations in fish liver and fish liver oils
Authors: La Frano Michael , Neidlinger Terry R , Patil Kavita , Burri Betty J
Published: June 2021

Fish, and especially fish liver oils, are known as excellent sources of vitamin A (VA). While it has long been recognized that fish livers contain VA in forms of retinol (ROH) and retinal (RA), some fish synthesize and use a VA‐like product (3‐hydroxy‐retinol; 3OHVA). Despite its possible presence, previous measurements of 3OHVA concentrations in fish liver are rare, and most of the data available is limited to fresh water species from India and Bangladesh. Meanwhile, there is a significant lack of knowledge regarding North American native fish sources of 3OHVA, a nutrient with potential health benefits. We have developed a reversed‐phase HPLC method for analyzing VA and 3OHVA in fish liver and fish liver oils. The method is a YMC Carotenoid column, 25cm, C‐18 gradient method varying from 100% acetonitrile to 80:20 acetonitrile:water using a diode array detector. Reproducibility ranged from 3–5% 3OHVA, 2–14% ROH, and 4–5% RA. Retinoid concentrations ranged widely, from <1–53 ug/g 3OHVA, 6–381 ug/g ROH, and 19–1794 ug/Kg RA. Tilapia, silver carp, catfish, and rainbow trout had varying amounts of 3OHVA, but it was not detected in the bass or steelhead. All fish contained ROH and RA.

Western Human Nutrition Research Center, USDA, supported this research.
Omega‐3 fatty acids and lipoprotein associated phospholipase A2 (Lp‐PLA2) in healthy older adult males and females
Authors: Nelson Tracy L , Hokanson John E , Hickey Matthew S
Published: June 2021

Background and Aims

Lipoprotein associated phospholipase A2 (Lp‐PLA2) is a novel inflammatory factor that has been independently associated with stroke and CVD. Omega‐3 fats have been implicated in reducing inflammation associated with CVD. The aim of this study was to determine if an 8‐week isocaloric diet supplemented with eicosapentaenoic acid (EPA) and docosahexaenoic (DHA) or α‐linolenic acid (ALA) would alter Lp‐PLA2 among healthy adults ages 50 years and older.

Methods and Results

Fifty‐nine healthy adults (75% female, average age 61 years) were randomized to one of three groups: the control group (n=19) who consumed olive oil capsules; the ALA group (n=20) who consumed flaxseed oil capsules and the EPA/DHA group (n=20) who consumed fish oil capsules. Fasting blood samples were obtained before and after the 8‐week intervention for determination of Lp‐PLA2 mass and activity. We did not find any significant changes in Lp‐PLA2 mass or activity in any of the groups; however, change in oxidized LDL was associated with change in Lp‐PLA2 mass (r=0.37, p<0.01).

Conclusion

Supplementing the diet with omega‐3 fatty acids for 8‐weeks did not influence Lp‐PLA2 activity or mass among older adults; altering oxidized LDL levels may be necessary to see changes in Lp‐PLA2 levels.

Supported by USDA, Agriculture Experiment Station, CSU
Whole grain and dietary fiber intake and prostate cancer aggressiveness by race
Authors: Tabung Fredoline , Steck Susan E. , Su L. Joseph , Arab Lenore , Fontham Elizabeth T.H. , Bensen Jeannette , Hebert James R. , Zhang Hongmei , Mohler James L.
Published: June 2021

Dietary fiber and whole grains are implicated in prevention of cancer, but have been studied rarely with regard to prostate cancer aggressiveness. The associations among intake of whole grains, refined grains and total dietary fiber and aggressiveness of prostate cancer were examined among African Americans (AA, n=667) and Caucasian Americans (CA, n=820) in a population‐based, case‐only study (the North Carolina‐Louisiana Prostate Cancer Project). Highly aggressive cases were defined as Gleason sum ≥8, or PSA>20ng/ml or Gleason sum=7 and stage cT3‐cT4, and low/intermediate aggressive cases were defined as all others. Logistic regression was utilized to test for associations of prostate cancer aggressiveness with intake of refined grains, whole grains and dietary fiber, stratifying by race. There was a non‐significant positive association between refined grains and risk of aggressive prostate cancer for AA (OR, 1.52; 95% CI, 0.82–2.82 for the 4th quartile compared to the 1st quartile of intake). A borderline significant inverse association between whole grain intake and prostate cancer aggressiveness was observed among CA (OR, 0.52; 95%CI, 0.27, 1.00). No other significant associations were observed. These results suggest that refined grain, whole grain and fiber intake are unlikely to be related substantially to prostate cancer aggressiveness.

Grant Sponsor: Department of Defense “DAMD 17‐03‐2‐0052”
Tumor progression locus 2 deletion attenuates alcohol‐induced hepatic inflammatory foci and cytokine expression in mice
Authors: Peach Camilla Anne , Hussain Sajid , Liu Chun , Wang Xiang-Dong , Greenberg Andrew S
Published: June 2021

The pathogenesis of alcohol‐induced fatty liver disease involves the interaction of several inflammatory signaling pathways. Tumor progression locus 2 (TPL2) is a kinase that functions as a critical regulator of inflammatory pathways by up‐regulating production of inflammatory cytokines. The present study aims to fill the gap in knowledge regarding the involvement of TPL2 in the mechanism of alcohol‐induced hepatic inflammation. In this study, male TPL2 knockout (TPL2KO) mice and wild‐type (WT) mice were pair‐fed with Lieber‐DeCarli liquid diet (LD) or alcoholic liquid diet (ALD, 27% energy from ethanol) for 4 weeks. Results show that the deletion of TPL2 significantly reduced inflammatory foci in the liver of mice consuming both LD and ALD as compared to their respective WT controls. This reduction was associated with suppression of hepatic inflammatory gene expression of interleukin‐6, tumor necrosis factor‐α, and interleukin‐1β. Histological analysis of livers showed TPL2 deletion significantly decreased steatosis in mice fed LD, but not in mice fed ALD, as compared to their respective WT controls. The demonstration that TPL2 deletion attenuates alcohol‐induced hepatic inflammation provides evidence for a novel role for TPL2 in the pathogenesis of alcohol‐induced fatty liver disease.

Grant Funding Source : NIDDK RO1‐DK‐082574/USDA 1950‐51000‐064S
Patterns and mechanisms of air flow during abdominal pumping in the grasshopper, Schistocerca americana
Authors: Harrison Jon Fewell , Cease Ariane J. , House Adam
Published: June 2021

While it has long been clear that convection (not diffusion) is the dominant mechanism for gas exchange in grasshoppers and other large insects, the patterns and mechanisms of air flow within insects remain poorly understood. Based on prior work by Weis‐ Fogh, Miller and Harrison it is believed that air flow in resting grasshoppers enters via the thoracic spiracles, flows longitudinally through the body along the gut, heart, nerve cord and spiracles, and exits primarily via the most terminal spiracles. We used synchrotron x‐ray imaging, high speed video and direct observation of spiracles to understand the patterns and mechanisms of abdominal air flow in the American locust, Schistocerca americana. We found that the abdomen contains collapsible air sacs and tracheae, as well as noncollapsible tracheae. During abdominal pumping, the viscera move anteriorly relative to the spiracles, contributing to compression of the air sacs and some of the tracheae, with others remaining open to allow air flow. In contrast to prior reports, the most caudal spiracles are mostly closed, opening briefly before air sac compression. An anteriorly‐directed compression wave and the timing of opening of the spiracles suggests that expiratory air flow is primarily forward‐directed within the abdomen, exiting via the first abdominal spiracles. Funded by NSF 0938047.
Decreased tight junction gene expression in the duodenum following heat stroke in F344 rats
Authors: Helwig Bryan G , Ward Jermaine A. , Leon Lisa R
Published: June 2021

Heat stroke (HS) morbidity and mortality is initiated by endotoxin escape (EE) from the intestinal tight junction barrier (TJB) although in vivo the molecular mechanisms of EE are not well understood. The current study tested the hypothesis that in the duodenum of male Fischer 344 (F344) rats tight junction gene expression is decreased following HS. F344 rats (276.6 gms) were heated until a maximum core temperature (Tc,Max; radiotelemetry ±0.1°C) of 41.8°C and allowed to recover for one, two or ten days. Duodenum gene expression of tight junction protein 1 (−2.06 fold), occludin (−2.13 fold), claudin 2 (−3.86 fold) and claudin 3 (−2.22 fold) was significantly decreased at Tc,Max (n=7; p<0.05) versus non‐heated controls (n=7). Claudin 2 gene expression was the only TJB protein that remained significantly decreased at one day (n=7;−2.93 fold, p<0.05) when compared to controls (n=8; p<0.05). At two and ten days gene expression of tight junction proteins was not different from control animals. These data suggest that decreased expression of TJB proteins may facilitate EE during the first 48h following HS. Research supported by MRMC. Author views not official US Army or DoD policy.
Involuntary exercise is a better stimulus than voluntary exercise to increase GDNF protein content in adult rat spinal cord
Authors: McCullough Monica Janine , Gyorkos Amy M , Spitsbergen John M
Published: June 2021

Glial cell line‐derived neurotrophic factor (GDNF) is the most potent neurotrophic factors for motor neurons. We have previously shown that short term involuntary exercise alters GDNF protein content in rat skeletal muscle, yet, little is known about how exercise regulates GDNF in the spinal cord of healthy individuals. The aim of the current study was to examine changes in GDNF protein content in the spinal cord from adult rats following involuntary and voluntary exercise of different durations. Male Sprague Dawley rats aged 6 to 24 months were examined. Exercised animals were placed into groups of either voluntary running wheels, involuntary running wheels, or forced swimming. Age‐matched control animals had no exposure to exercise. GDNF protein content of the spinal cord was measured using an ELISA and western blot. Immunohistochemistry localized GDNF and examined motor neuron morphology. Two weeks of involuntary running resulted in the greatest change in GDNF protein content (6.5‐fold increase) whereas 6 months of voluntary running decreased GDNF protein content (0.8‐fold decrease). Short‐term involuntary exercise may be better than long‐term voluntary exercise to increase GDNF in the spinal cord of aging individuals and provide neuroprotection of the motor nervous system with aging. This work was supported by NIH grant 1 R15 AG022908‐ 01A2, NSF grant DBI 0552517, Western Michigan University, and MSU‐KCMS.
Controlling proteome degradation in Daphnia pulex
Authors: Kemp Catherine Jarrett , Kültz Dietmar
Published: June 2021

Previous two‐dimensional gel electrophoresis (2DGE) studies in Daphnia pulex have described uncontrolled, widespread proteome degradation. An attempted 2DGE study of the D. pulex response to sodium chloride exposure revealed an unexpected result: the previously observed proteolysis is halted following exposure of animals to elevated salinity prior to protein extraction. Further investigation of conditions controlling proteome degradation shows that the proteolytic factor(s) is/are present in an active form in extract from control “normal/healthy” animals; can be negatively regulated by prior in vivo salinity exposure of animals or inhibited by post‐homogenization boiling of samples; and retain proteolytic activity under chemically denaturing conditions. Extended characterization of the environmental and biochemical parameters that influence this degradation are described. Funding provided by NIH grant 5 P42 ES004699‐25, NSF grant IOS‐ IOS‐1049780, UC‐Davis Floyd and Mary Schwall Fellowship in Genetics, UC‐Davis Jastro‐Shields Fellowship.
Pregnancy status and choline intake alter DNA integrity, epigenetic marks and gene expression
Authors: Jiang Xinyin , Yan Jian , West Allyson , Perry Cydne , Malysheva Olga , Bar Haim , Wells Martin , Devapatla Srisatish , Pressman Eva , Caudill Marie
Published: June 2021

This study sought to systemically elucidate the effects of pregnancy and maternal choline intake on genomic markers. Healthy third trimester pregnant (n=26, wk 27 gestation) and non‐pregnant (n=21) women were randomized to controlled choline intakes of 480 or 930 mg/day for 12‐wk. Leukocyte samples were acquired at wk 0 and 12. Pregnant women had higher DNA damage compared to non‐pregnant women (P < 0.001), suggesting insufficient defense against oxidative stress. Genome‐wide transcript profiling revealed the up‐regulation of genes related to immune response, consistent with an activated host defense system towards the end of pregnancy. Pregnant women responded to the higher choline intake with increased (P < 0.05) histone 3 lysine 4 di‐methylation (H3K4me2), a marker of transcription activation. In sum, pregnancy altered markers of genomic stability and modulated the expression of immune‐related genes in leukocytes. The epigenetic modification of histones among pregnant women consuming varied choline intakes may have downstream effects on chromatin restructuring and gene transcription.

Grant Funding Source: Funded in part by USDA, ENC and the Beef Checkoff through NCBA