bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel surgical technique for direct substance or cell delivery into the cerebral vasculature
Authors: Chen Lei , Swartz Karin R. , Toborek Michal
Published: June 2021

Cerebrovascular research suffers from a lack of reliable methods to effectively deliver exogenous substances into the central nervous system (CNS) of small experimental animals. In this study, we describe a novel vessel microport surgical technique for a variety of cerebrovascular applications that is reproducible and well tolerated in mice. The procedure is based on the insertion of a vessel microport into the external carotid artery for substance delivery into the CNS via the internal carotid artery. The method results in selective substance delivery into the ipsilateral hemisphere. Other novel aspects of this surgical technique include the ability to perform multiple injections, study of conscious mice well‐removed from surgery, and the lack of occlusion of the common or internal carotid artery that allow maintaining of carotid flow. The feasibility of this technique has been validated by infusion of HIV Tat protein to induce permeability of the blood‐brain barrier and by implantation of tumor cells to establish a brain metastasis model. Thus, the described vessel microport technique can be employed in a variety of cerebrovascular research applications. Supported by the NIH grants: MH 63022, MH 072567, NS 39254, and P42 ES 07380
Functional characterization of UbcH7 interaction with the E6AP ubiquitin ligase
Authors: Ronchi Virginia Paola , Haas Arthur L
Published: June 2021

E6AP is the founding member of the HECT domain superfamily of ubiquitin ligases, mutations in which are responsible for Angelman syndrome. Kinetic analysis of polyubiquitin chain formation shows UbcH7 is the functionally cognate E2 for E6AP, the concentration dependence of which yields a Km=58±6 nM and kcat=0.013 s−1. Other kinetic studies indicate E6AP harbors two UbcH7 binding sites: Site 1 involved in the initial formation of the Hect domain~Ub thiolester and Site 2 required for chain elongation. Mutations designed to disrupt Site 2 interactions predicted from the crystal structure support the model. UbcH7K100A shows nearly indistinguishable kinetics with Km= 31 ± 17 nM and kcat=0.081±0.008 s−1, indicating Site 1 using binding interactions distinct from Site 2. UbcH7F63A forms the initial Hect~Ub thiolester but doe not support chain elongation even though it binds E6AP and is a competitive inhibitor of Site 1 bound UbcH7 (Ki =410 nM). Pulse‐chase kinetics with 125I‐ubiquitin confirms chain elongation occurs by distal addition to the growing polyubiquitin chain. RAD23A, substrate of E6AP, negatively affects the polyubiquitination of substrates by Ubc2. The kinetic analysis shows that RAD23A was an uncompetitive inhibitor of the conjugation reaction catalyzed by UbcH7‐E6AP (Ki 414 nM). These studies reveal the complex mechanism of interaction among UbcH7, E6AP and RAD23A. (Supported by GM340092 to A.L.H.).
The molecular basis for transmission of human prion strains
Authors: Giles Kurt , Patel Smita , De Nicola Gian , Korth Carston , Lemus Azucena , DeArmond Stephen , Prusiner Stanley B
Published: June 2021

Human prion diseases, such as Creutzfeldt‐Jakob disease (CJD), can occur in different "strains" which are characterized by their clinical onset, and by biochemical and neuropathological hallmarks of the post mortem brain. Human prion strain type is dramatically influenced by the M/V polymorphism at residue 129, with homozygous individuals much more likely to develop disease.

We report a series of novel transgenic (Tg) mouse lines expressing various chimeras of mouse and human prion protein (PrP), which were inoculated with either of the two most common strains of sporadic (s) CJD, termed MM1 and VV2, or with the variant (v) CJD strain. The differential susceptibility for each strain allows a molecular dissection of the amino acid residues that influence strain propagation.

One Tg line was susceptibility to sCJD(MM1) prions in less than 80 days, and defined six human PrP residues in the central region of the molecule that are essential for strain transmission. Conversely, for the sCJD(VV2) strain, transmission is regulated by residues in the terminal regions of the PrP sequence. Modification of strain characteristics on transmission of sCJD(VV2) and vCJD strains, also allowed us to formulate a general set of rules governing strain transformation.
CD36 Mediates a Pro‐inflammatory Signaling Loop in Fat and Contributes to Insulin Resistance
Authors: Kennedy David J , Silverstein Roy L , Febbraio Maria
Published: June 2021

Scavenger receptor CD36 recognizes ligands generated in pro‐atherogenic hyperlipidemic (PA‐HL) states, including oxidized LDL (oxLDL). Systemic levels of oxLDL are highly associated with insulin resistance (IR). We hypothesized that CD36 ligands generated in PA‐HL states disrupt insulin signaling in adipocytes via CD36‐dependent signaling.

CD36+ and CD36− mice were fed a high‐fat diet for 8 weeks and examined for markers of IR. Compared to CD36+, CD36− mice had improved glucose tolerance and both decreased JNK and inhibitory IRS‐1 serine phosphorylation in adipose tissue. Further, macrophages from CD36− mice demonstrated decreased production of ROS and proinflammatory cytokines, and increased arginase activity compared to cells from CD36+ mice.

L1 adipocytes treated with oxLDL showed time and dose dependent increases in CD36, activated JNK, inhibitory IRS‐1 serine phosphorylation, and adipocyte lypolysis with decreased glucose uptake. Pre‐incubation with either a CD36 mAb or JNK inhibitor attenuated these changes. In addition, adipocytes co‐cultured with transwell separation from CD36− macrophages showed both decreased JNK and inhibitory IRS‐1 serine phosphorylation compared to those co‐cultured with CD36+ macrophages.

These data suggest a CD36‐dependent paracrine loop between adipocytes and macrophages that facilitates chronic inflammation in adipose and underlies IR common to PA‐HL states.
The Protein Structure Initiative Structural Genomics Knowledgebase
Authors: Gabanyi Margaret , Westbrook John , Tao Wendy , Shah Raship , Kouranov Andrei , Schwede Torsten , Arnold Konstantin , Kiefer Florian , Bordoli Lorenza , Podvinec Michael , Kopp Jurgen , Adams Paul , Carter Lester , Minor Wladek , Nair Rajesh , La Baer Joshua , Berman Helen M.
Published: June 2021

The Protein Structure Initiative (PSI) has produced more than 3300 protein structures in a high throughput manner. In order to achieve this, new technologies for all aspects of the structure determination and analysis pipeline have been developed. The strategies used for target selection by PSI centers have resulted in the structural determination of many novel sequences, and an increase in the structural coverage of protein families.

The PSI Structural Genomics Knowledgebase (PSI SGKB) gives centralized access to the structures, annotations, models, protein production protocols, materials and technologies catalyzed by the PSI efforts. In collaboration with the Nature Publishing Group, the PSI SGKB expanded to become the Structural Genomics Gateway that provides a research library, editorials about new research advances and technologies, news, and an events calendar to present a broader view of structural biology and structural genomics. A description of how this resource can be used for enabling biological research will be given.

The PSI SGKB is funded by the NIGMS.
Functional analysis of yeast fRMsr and its role in the reduction of free methionine‐R‐sulfoxides in yeast and mammalian cells
Authors: Lee Byung Cheon , Marino Stefano M , Le Dung Tien , Zhang Yan , Gladyshev Vadim N
Published: June 2021

Methionine sulfoxide reductases (Msrs) are oxidoreductases that catalyze thiol‐dependent reduction of oxidized methionines. MsrA and MsrB are well known Msr enzymes, which have reduce methionine‐S‐sulfoxide (Met‐S‐SO) and methionine‐R‐sulfoxide (Met‐R‐SO), respectively. Recently, an additional Msr specific for free Met‐R‐SO from an Escherichia coli was discovered. Here, we characterized occurrence and function of fRMsr homologs via comparative genomics and experimental analyses in Saccharomyces cerevisiae. We also propose a reaction mechanism of this enzyme based on an enzyme‐substrate docking model using computational molecular dynamics and on the experiments involving trapping fRMsr‐based intermolecular intermediates with thioredoxin 1. In addition, transfection of fRMsr into mammalian SK‐Hep1 cells increased oxidative stress resistance and restored deficiency of these cells in the reduction of free Met‐R‐SO. Altogether, the data shows yeast fRMsr is the major protein that reduces free Met‐R‐SO in yeast cells and can also support this function in mammalian cells.
Host SNAREs mediate fusion of vacuoles containing Legionella pneumophila with vesicles exiting the endoplasmic reticulum
Authors: Arasaki Kohei , Roy Craig
Published: June 2021

Legionella pneumophila is an intracellular pathogen that initially resides in a plasma membrane‐derived vacuole that fuses with endoplasmic reticulum‐derived vesicles to create a specialized organelle that supports bacterial replication. Host SNARE proteins were examined to determine the mechanism of fusion of this vacuole with early secretory vesicles. Localization studies determined that plasma membrane t‐SNARES are associated with vacuoles containing Legionella shortly after bacterial uptake into mammalian cells. Syntaxin‐2, Syntaxin‐3, Syntaxin‐4, and SNAP23 were all localized to vacuoles containing Legionella. The v‐SNARE Sec22b was rapidly recruited to vacuoles containing Legionella after internalization. Reducing production of Syntaxins by RNAi inhibited the recruitment of Sec22b to vacuoles containing Legionella. Infection by Legionella induced the formation of SNARE complexes containing plasma membrane Syntaxins, SNAP23, and Sec22b. Syntaxin, SNAP23, and Sec22b complexes formed during infection were dissociated upon the addition of NSF/α‐SNAP, indicating that these are functional SNARE complexes. Thus, Legionella has the ability to manipulate the pairing of host SNARE proteins to efficiently promote fusion of a plasma membrane‐derived vacuole with early secretory vesicles. NIH grant R01 AI041699 supported this work.
Bacterial Acyl Carrier Protein Interacts with the Alarmone SpoT and the Adaptive Response Protein AidB: Roles for Acyl Carrier Protein Beyond Lipid Metabolism
Authors: Cottle Julia L , McMaster Christopher R , Byers David M
Published: June 2021

Acyl carrier protein (ACP) plays an essential role in the synthesis of bacterial lipids, and is an attractive target for the development of new antibiotic classes. ACP also forms numerous interactions with proteins involved in other cellular processes that have yet to be fully identified or characterized. Using a C‐terminal Tandem Affinity Purification (TAP)‐tagged ACP we identified two ACP binding partners not directly involved in lipid metabolism, AidB and SpoT, that both exhibit increased interaction with ACP in stationary phase. The increase in binding of SpoT, a synthetase and hydrolase of the alarmone guanidine ‐ 5' (tri)diphosphate ‐3'diphosphate, to ACP appears to be due to increased affinity between the two proteins as cells enter stationary phase. In contrast, the increased interaction with AidB, an adaptive response protein, results from elevation in the level of AidB as cells enter stationary phase. AidB levels are also increased upon exposure of cells to alkylating agents. AidB binds DNA directly and we hypothesize that the positively charged DNA binding region of AidB also binds to the negatively charged ACP. AidB binding to ACP may have important functional consequences in DNA repair. To test this, ACP‐AidB protein‐protein interaction domains are being mapped, and we are determining if AidB‐ACP interactions regulate the ability of the cell to respond to DNA damaging agents. (Supported by NSERC)
Heterogeneity in platelet secretion
Authors: Jonnalagadda Deepa , Whiteheart Sidney W.
Published: June 2021

Platelets, especially their α‐granules, are repositories for many cytokines and growth factors and thus, have profound roles in thrombosis, inflammation, atherosclerosis, and angiogenesis .Our work seeks to determine if these platelet cargo molecules are released with differential kinetics and/or in response to different agonists. Our experiments use antibody arrays to quantify the release of ~20 different proteins. In the initial stages of our analysis, thrombin has been used as agonist (activation through PAR receptors) and platelets have been stimulated for increasing times. We have detected two different classes of secretion from α‐granules that could be categorized as slow and rapid secretion events. These kinetic studies of secretion from α‐granules are being done in conjunction with dense core granule and lysosomal granule release and the pattern followed by the α‐granules is intermediate, with densecore granule release being very rapid and lysosomal granule release being much slower. Similar kinetics could be seen when they were stimulated with Convulxin (activation through GPVI pathway) and PAR1 activation peptide. Future experiments will determine if other agonists alter the pattern and/or kinetics of platelet cargo release. This work is supported by NIH grants (HL56652 and HL091893) to S.W.W.
Characteristics of short gel filtration columns for rapid screening of aggregates and impurities
Authors: Heijbel Anna , Salomonson Inger , Zuo Jinyu , Carlsson Marianne , Lundstrom Fredrik
Published: June 2021

Gel filtration (GF) is a gentle technique suitable for the study of aggregation and purity of a target protein. GF, however, is often time consuming and can also be held up due to the expense of sample and buffers. A new, small (3 ml) column format packed with Superdex(tm) 75 and Superdex 200 has been developed for rapid and reliable gel filtration enabling analyses in less than 6 min using only 4 µl of sample.

Results illustrating the resolution of a standard protein mixture, sensitivity, flow rate properties and influence of system dead volumes upon resolution will be shown. The short gel filtration columns will also be compared to the longer Superdex 75 and 200 10/300 GL columns.
Determining The Role of Subcellular Localization of Heterotrimeric G Protein βγ Subunits in a Trans Golgi Network‐To‐Plasma Membrane Vesicle Formation Pathway
Authors: Irannejad Roshanak , Wedegaertner Philip B
Published: June 2021

Heterotrimeric G proteins are responsible for transducing signals initiated at the plasma membrane (PM) by G protein‐coupled receptors (GPCR). However, a number of recent studies indicate that G proteins can signal at endomembranes. Gβγ subunits have been implicated in activating protein kinase D through PKCη at the Golgi membrane to initiate a pathway causing fission of PM‐destined vesicles from Trans Golgi Network (TGN). Overactivation of these signaling components cause the complete fission of the Golgi, while inhibition of some of the components of this pathway decrease TGN‐to‐PM transport of proteins. What is not clear is the intracellular location(s) where Gβγ initiates this signaling pathway. Our data indicate that constitutively targeted Golgi‐βγ, but not PM‐, ER‐ and cytosolic‐βγ, induces complete fission of Golgi as visualized by fluorescent microscopy. Moreover, rapamycin‐inducible recruitment of Gβγ to the Golgi but not PM activates the TGN‐to‐PM vesicle formation pathway. Sequestration of Gβγ with the C‐tail of GPCR kinase 2 (GRK2ct) blocks the transport of two model proteins, GFP‐tagged vesicular stomatitis virus‐G protein (GFP‐VSV‐G) and HRP‐tagged N‐terminal signal sequence (ss‐HRP). The results identify a role for endogenous Gβγ at the Golgi in regulating vesicle transport.

[Supported by NIH GM56444 and AHA 0815431D]
Caspase‐3 digestion of cortactin: Analysis of Cortactin Structure
Authors: Foster Jeneva , Kruchten Anne E
Published: June 2021

Cortactin is a Src substrate protein that interacts with actin, and is involved in several types of cancer metastases. Phosphorylation of cortactin by Src and Erk affects its role in cancer metastases, possibly via induced changes in the folding of the protein. To test if phosphorylation alters the structure of the protein, wild type and two mutants (one to mimic, and one to block phosphorylation) were digested in‐vitro with the protease caspase‐3, which has two potential target sites on cortactin. Cortactin was digested at varying times and with varying concentrations of caspase‐3 in reaction; results showed that the wild type and mutant proteins were digested at varying rates. We hypothesize variation in digestion is a result of different folding patterns between wild type and mutants due to either an exposed target site or a target site buried within the structure of the protein. This work was supported by a Faculty Start Up Grant to A.E. Kruchten by The Murdock Charitable Trust and by a Linfield College Faculty Student Collaborative Research Award to A.E. Kruchten and J. Foster.
Analysis of the structure of cortactin by native gel electrophoresis
Authors: Petrovich Jessica , Kruchten Anne E
Published: June 2021

Cortactin is an actin‐binding protein that acts as a Src‐kinase substrate and has been shown to be involved in cellular migration and metastases. Previous studies have shown that cortactin is an elongated protein with limited tertiary structure. It is understood that phosphorylation of cortactin affects its role in cellular motility and metastases but the mechanism for this remains unknown. We hypothesize that phosphorylation induces a conformational change in the folding pattern of cortactin. Non‐denaturing acrylamide electrophoresis gels (native gels) were run of mutant phosphorylation‐inhibiting and mocking mutants of cortactin and compared to wild type. We found differences in the migration of bands between the wild type and mutant forms, suggesting conformational differences between them. This work was supported by a Faculty Start Up Grant to A.E. Kruchten by The Murdock Charitable Trust and by a Linfield College Faculty Student Collaborative Research Award to A.E. Kruchten and J. Petrovich.
Analysis of RET co‐receptor mutants in receptor activation
Authors: Vega Quinn C.
Published: June 2021

RET is a mammalian receptor tyrosine kinase required for both proper neuronal and kidney development. Mutations in the ret gene are associated with Multiple Endocrine Neoplasia (MEN) 2A, MEN 2B and Familial Medullary Thyroid Carcinoma (FMTC) as well as Hirschsprung's Disease, a disease characterized by disruption in neuronal development. Wild type RET is activated after association with a membrane associated co‐receptor GFR‐α (1‐4) and its corresponding ligand (GDNF, Neurturin, Artemin or Persephin). In order to further understand the mechanism by which the co‐receptors activate RET, mutations were made in GFRa‐1 and the resulting mutants analyzed for their ability to activate RET. Mutations were made based on previous studies and through a comparison of co‐receptor sequences. Using a RET dependent luciferase reporter gene assay, specific co‐receptor mutants were transfected into NB41A3 cells and RET activity measured in the presence or absence of GDNF. Although wild type GFRα‐1 was able to increase luciferase activity in the presence of GDNF, individual co‐receptor mutants could not activate the receptor. Specifically, point mutations and insertion mutations in the center of the co‐receptor were able to disrupt co‐receptor activity. This decrease in activity was not due to decreased expression of the co‐receptor.
Role of Caveolin‐1 in Gastrointestinal Smooth Muscle Function
Authors: Clay Chereena T , Mahavadi Sunila
Published: June 2021

The objective of this study was to determine the role of caveolin‐1 in smooth muscle function using colonic segments from wild type and caveolin‐1 knock out (caveolin‐1‐/‐) mice. When conducting this study colonic segments from four wild type and four caveolin‐1‐/‐ mice were isolated and suspended in an organ bath containing 15 ml of Krebs solution. Isometric contractions were recorded by a force transducer connected to a personal computer using Acqknowledge 382 software program. Contraction in response to muscarinic agonist was tested using different concentrations of carbachol (10 nM‐300 mM), and relaxation in response to soluble guanylyl cyclase activator and adenylyl cyclase activator was examined using sodium nitroprusside (SNP: 100 nM to 1 mM) and forskolin (10 µM), respectively. Contraction was measured as increase in tension in grams and relaxation was measured as a percentage of the decrease in tension from the carbachol‐induced contraction. The smooth muscle responses in the colon of the caveolin‐1‐/‐ mice where then compared to those in the colon of the wild type mice. We hypothesized that smooth muscle function would be altered in the caveolin‐1‐/‐ mice. Our studies showed treatment with carbachol caused a concentration‐dependent contraction of colon in both wild type and caveolin‐1‐/‐ mice. The magnitudes of contraction in wild type and caveolin‐1‐/‐ mice were not significantly different. Addition of forskolin or SNP caused relaxation of the colonic muscle contracted with carbachol in both wild type and caveolin‐1‐/‐ mice. The response to forskolin was not significantly different, whereas the responses to SNP were reduced in colonic segments from caveolin‐1‐/‐ mice compared to wild type mice. We conclude that caveolin‐1 plays an important role in smooth muscle relaxation mediated by soluble guanylyl cyclase/cGMP pathway.
[beta]‐Carotene content and bioaccessibility from organic and non‐organic baby meals
Authors: Aherne‐Bruce Aisling , Jiwan Marvin A , Duane Paula , O'Sullivan Laurie , O'Brien Nora M
Published: June 2021

Research data on the nutritional benefits to babies of consuming organic foods compared with their non‐organic counterparts is scarce. The objectives of the present study were first, to determine the β‐carotene content of organic and non‐organic baby food products; and, second, to compare the bioaccessibility of β‐carotene from these foods. Carotenoid bioaccessibility (%) is defined as the proportion of ingested carotenoid(s) that are available for absorption in the gut after digestion. Three organic (A‐C) and three non‐organic (D‐F) chicken and vegetable baby foods for babies of 4+ months of age were subjected to a simulated human gastrointestinal digestion procedure. Within each group (i.e. organic or non‐organic), there were large differences in β‐carotene content. For example, β‐carotene content of the organic dinners ranged from 8.8 to 534.9 μg/100 g. Similarly, non‐organic dinner F had significantly greater amounts of β‐carotene than the other t+wo non‐organic dinners tested. There was no difference in the average β‐carotene content between the organic and non‐organic foods. β‐Carotene bioaccessibility was greater from organic dinner C (46.4%) compared with organic dinners A and B (P<0.05). On average, β‐carotene had greater % bioaccessibility (P<0.05) from the non‐organic (41.4%) than organic (30.8%) chicken and vegetable dinners. Our findings show that β‐carotene content reflected the vegetable composition of the baby meals. To conclude, the organic dinners tested were not superior to the non‐organic foods in terms of β‐carotene content and bioaccessibility.

This research was funded by Science Foundation Ireland.
Induction of apoptosis by L‐carnitine through activation of two main pathways in Hepa1c1c 7 cells
Authors: Han Gi Dong , Chung Hyun Chae , Kim Han‐Sup , Kim Dong Yeop , Ko You Hyok , Jeong Bo‐Young , Fan Jiang Ping
Published: June 2021

This study shows the effects of L‐carnitine treatment on cell proliferation with hepa1c1c7 mouse cancer cells and NCTC 1469 normal cells. In an MTT assay, L‐carnitine increased the number of dead hepa1c1c7 cells, while there was no difference in the number of NCTC 1469 cells. mRNA and protein levels of TNF‐α, Fas, and caspase‐8, which are closely related to cell apoptosis by a death ligand/receptor‐dependent apoptosis pathway, were increased by L‐carnitine treatment. In addition, L‐carnitine treatment activated mitochondria‐dependent apoptosis pathways by inducing the up‐regulation of caspase‐9 and caspase‐3 and the down‐regulation of Bcl‐2 in hepa1c1c 7 cells. Taken together, the findings of this study have demonstrated that L‐carnitine could induce apoptosis in hepa1c1c7 cells by activating Fas lagands and inhibiting the expression of Bcl‐2. These results suggest that L‐carnitine treatment could be related to both a mitochondrion‐dependent and a death ligand/receptor‐dependent apoptosis pathway in hepa1c1c7 cells. Our results could give information for understanding the L‐carnitine‐induced apoptosis mechanism in cancer cells.
An elevated protein diet does not accelerate initiation and early progression of MNU‐induced breast cancer tumors in rats
Authors: Moulton Christopher John , Valentine Rudy , Wallig Matthew , Monaco Marcia , Singletary Keith , Layman Donald
Published: June 2021

There is growing interest in identifying nutritional factors that modify risk and development of breast cancer. This study evaluated the influence of feeding low vs. high protein diets (LP vs. HP) on measures of tumor incidence. LP and HP diets consisted of 60:15:25 (carbohydrate:protein:fat as % energy) and 40:35:25, respectively. Female SD rats ~140 g were injected intraperitoneally with N‐nitroso‐N‐methylurea (MNU) at 50 mg/kg, and subsequently fed 3 times/d for 10 wk to mimic human meal patterns. Body weight, food intake and tumor appearance/growth were monitored throughout the study. At 10 wk, rats were killed; serum insulin and IGF‐1, tumor size and pathology were evaluated. Food intake did not differ between groups but final body weight was 4% greater in HP. No differences existed in fasted or post‐prandial insulin, however post‐prandial IGF‐1 was significantly greater in HP. There were no differences in tumor latency, cumulative surface area, mass or stage of histological development. Although more LP animals (56%) had tumors than HP (40%), the difference was not significant (p = 0.19). Despite elevation of IGF‐1 in HP, this study fails to demonstrate any direct effect of normal levels of dietary protein or carbohydrate to exacerbate the initiation and early progression of breast cancer tumors.

Support: Illinois Council on Food and Agricultural Research
Urinary phytoestrogen excretion and prostate cancer risk in a nested case‐control study: The Multiethnic Cohort
Authors: Park Song‐Yi , Wilkens Lynne R. , Franke Adrian A. , Le Marchand Loïc , Kakazu Kerry K. , Goodman Marc T. , Murphy Suzanne P. , Henderson Brian E. , Kolonel Laurence N.
Published: June 2021

Phytoestrogens are of special interest in prostate cancer research because populations in Asia with a high consumption of phytoestrogens have a lower incidence of the disease than comparable populations in Western countries. This case‐control study is nested within a multiethnic cohort in Hawaii and California. Analyses included 249 incidence prostate cancer cases and 404 matched controls for urinary daidzein, genistein, equol, and enterolactone. Conditional logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs) to account for the matching criteria as well as adjusted for covariates. The median excretion of daidzein was 0.173 nmol/mg creatinine in cases and 0.291 in controls (p = 0.01), and the median excretion of genistein was 0.048 in cases and 0.078 in controls (p = 0.05). An inverse association was seen for daidzein overall (OR for the highest vs. lowest quintile = 0.55, 95% CI = 0.31‐0.98, p for trend = 0.03) and for localized as well as advanced or high grade cancer and was suggested for each of the four ethnic groups examined. Urinary excretion of genistein, equol, and enterolactone was not significantly related to prostate cancer risk. Our findings suggest that high intake of phytoestrogens, as reflected by urinary excretion of daidzein, may be protective against prostate cancer. Grant support: P01 CA33619 and R37 CA54281.
The nutritional status and quality of life of home parenteral nutrition patients during one year of observation
Authors: Lysiak‐Szydlowska Wieslawa , Urbanowicz Krystyna , Malgorzewicz Sylwia
Published: June 2021

The aim was to determine the nutritional status and the quality of life of patients on home parenteral nutrition during one year of observation.

Methods

The group of 30 patients (17 F, 13 M, range of age 22‐85 years old) was analyzed. SGA (Subjective Global Assessment), BMI (Body Mass Index), TLC (total lymphocyte count) and plasma albumin concentration were used for nutritional assessment.

The quality of life was assessed by SF‐36 questionnaire and WHOQOL‐100 test.

Results

At the beginning of treatment one of the patients presented good nutritional status, 3 patients ‐ mild malnutrition, 13 patients severe malnutrition and 13 were cachectic. After one year of treatment an improvement of SGA scoring was observed in 77 % of patients; no change in 23 % of patients and no case of deterioration was noticed. Weight gain was observed in 30 % of patients; TLC showed improvement in 27% and decrease in 20% of cases.The increase of albumin concentration was observed in 40% of patients. After a year of parenteral nutrition 80% of patients declared an improvement in majority of domain of the SF‐36 questionaire and WHOQOL‐100 test. Awarness of "actual state of health" was significantly influenced by nutritional status (p<0.05).

Conclusions

The state of nutrition has been improved within the first year of parenteral feeding program. The quality of life is not a constant and permanent value but it is a dynamic process, variable in direction and dependent on an element of quality of life being considered individually.
The Legume Inflammation Feeding Experiment (LIFE): Effects of Gut Hormone Levels
Authors: Zhang Zhiying , Lanza Elaine , Bagshaw Deborah , Colburn Nancy , Ross A. Catharine , Rovine Michael , Hartman Terry
Published: June 2021

The LIFE Study was designed to test the effects of a legume‐rich diet on biomarkers of insulin resistance (IR) and inflammation. A secondary objective was to test the effect of the diet on gut hormones: leptin, ghrelin (total), and cholecystokinin (CCK). Sixty‐four men (35‐75 yrs), who had undergone a screening colonoscopy within the past 2 years, were recruited in Central Pennsylvania to consume a legume diet (LG,1 1/2 cups of cooked dry beans) and an isocaloric average American diet (AA) in random order each for 4 weeks separated by a 3‐wk compliance break. Preliminary results show that fasting plasma leptin concentrations were reduced on both LG (‐0.118 ± 0.026 S.D. ng/mL, P<0.0001) and AA diets (‐0.072 ± 0.019 ng/mL, P=0.0003) but the difference between diets was not statistically significant (P=0.1499). Fasting ghrelin levels slightly decreased on LG diet (‐15.289 ± 16.709 pg/mL) and increased on AA diet (19.129 ± 19.607 pg/mL); however, neither diet had an effect on ghrelin (P=0.3637 and P=0.3330, respectively). These results indicate that a legume‐rich diet may have favorable effects on lowering fasting leptin level with stable body weight. The measure and data analysis of CCK is ongoing. This study is supported by the National Cancer Institute and partial support provided by the GCRC, PSU.
Inhibitory effects of Aralia elata extract fractions on the biosynthesis of inflammatory mediators
Authors: Kim Gun‐Hee , Jeong Choon‐Sik , Lee Je‐Hyuk
Published: June 2021

Aralia elata (A. elata) has been used as a folk medicine for rheumatoid arthritis, diabetes, spasm of the stomach in Japan and Korea. Ethyl acetate‐fraction showed a scavenging activity of most DPPH free radical and reducing power of extract‐fractions showed the similar results. All A. elata extract fractions inhibited LPS‐induced nitrite production in a dose‐dependent manner. In the cell‐free system, all extract fractions chemically quenched NO partially, except aqueous fraction. Also, the iNOS mRNA transcription was suppressed in LPS‐stimulated RAW 264.7 cells. Especially ethyl acetate fraction (100 ¥ μg/mL) suppressed iNOS mRNA transcription to the basal level. In addition, all A. elata extract fractions showed the inhibitory effect on the production of prostaglandin E2 and suppressed the corresponding COX‐2 mRNA transcription, except chloroform fraction. The production of interleukin‐1¥â and ‐6 were inhibited by methanol extract, ethyl acetate fraction, and butanol fraction to almost basal level. Based on these results, A. elata methanol extract fractions may be useful as an alternative medicine for the relief and retardation of immunological inflammatory responses through the reduction of inflammatory mediators, including NO, PGE2, and cytokines.
Volatile Aroma Compounds of Atractylodes lancea DC.
Authors: Kim Gun‐Hee , Chang Kyung Mi
Published: June 2021

The volatile aroma constituents of Atractylodes lancea DC.were separated and extracted using a Clevenger‐type apparatus by steam distillation extraction method, and were analyzed by gas chromatography‐mass spectrometry (GC/MS). The yield of the essential oil from Atractylodes lancea DC.was 1.0% (v/w), and its color was the pale light yellow. Forty‐seven volatile flavor compounds, which make up 93.85% of the total volatile composition of the distilled oil were tentatively identified. It contained 35 hydrocarbons (81.25%) with sesquiterpene predominating, 5 alcohols (0.11%), 1 ketone (0.01%), 2 aldehydes (0.02%), 2 esters (0.16%), and 2 miscellaneous components (12.32%). Furanodiene (26.41%), 2,7‐dimethoxy‐3,6‐dimethylnaphthalene (13.56%), and 6,7‐dihydroxyxanthotoxin (12.30%) were the predominantly abundant aroma components in Atractylodes lancea DC., medicinal and aromatic plant.
Chromium histidinate improves serotonergic properties and carbohydrate metabolism in rat models of insulin resistance and of type 2 diabetes
Authors: Komorowski James Richard , Sahin Kazim , Tuzcu Mehmet , Sahin Nurhan , Akdemir Fatih , Juturu Vijaya
Published: June 2021

The present study evaluated the effects of chromium histidinate (CH) on serotonergic properties and carbohydrate metabolism. Insulin resistance and diabetes were induced in male, Wistar rats by feeding a high fat diet (HFD, 40% of calories as fat) for 2 weeks, administering a single injection of streptozotocin (STZ, 40 mg/kg i.p.), and continuing the HFD for another 10 weeks. Rats (10/group) were then divided into 6 groups (Control, Control+CH, HFD, HFD+CH, HFD/STZ, HFD/STZ+CH). The control group received a standard diet (12% of calories as fat) and the HFD and HFD/STZ groups continued receiving the HFD. Chromium (Cr) treated rats were fed CH (110 mcg/kg body wt/d). HFD and HFD/STZ treatment decreased brain Cr, tryptophan, serotonin, and serum melatonin (p<0.001) and increased blood glucose (p<0.01). CH treated rats (HFD+CH and HFD/STZ+CH groups) increased brain Cr, tryptophan, serotonin, and serum melatonin (p<0.05). The HFD/STZ group decreased blood glucose (p<0.001). In a second study using the HFD model, the addition of CH to chromium picolinate (CP) treated rats resulted in a faster and longer lasting reduction in blood glucose compared to CH or CP treatment alone. These results suggest that CH may be beneficial for correcting serotonergic defects and hyperglycemia caused by insulin resistance or diabetes, and that a hydrophilic Cr (CH) can enhance the benefits of a lipophilic Cr (CP).
Metrological tools to enhance the accuracy of copper, iron and zinc content of infant formula
Authors: Castanheira Isabel , Raimundo Inês , Santiago Susana , Sanches‐Silva Ana , Iyengar Venkatesh
Published: June 2021

Objective

To apply metrological tools such as Quality Control Materials (QCM) and Proficiency Testing (PT) schemes to enhance the reliability of the measurement process of Cu, Fe and Zn in food matrices.

To generate high quality analytical data to minimize errors by computing values for daily dietary intakes for specific population groups.

Materials and methods

Cu, Fe and Zn were determined in infant formula using atomic absorption spectrophotometric method (AOAC Official Method 985.35).

Results

The content of select minerals were determined after an independent method validation. A QCM was successfully prepared compiling with homogeneity and stability requirements. Traceability to basic SI units was achieved using Standard Reference Materials prepared by NIST. The participation in PT schemes revealed a consistent good laboratory practice. According to the results, the levels of Cu, Fe and Zn in infant formula were 0.28‐0.36, 5.79‐6.24, and 3.49‐3.91 mg/100g infant formula, respectively.

Conclusions

Due to the unequivocal role of Cu, Fe and Zn in the biological systems, it is important to know the accurate levels of these minerals in foodstuffs. The use of QCM and PT schemes allow guaranteeing data quality, which is a crucial issue to monitor food/ health relationship.

Funding Disclosure

Project LMARSA POR/3.1.015/DRELVT and European Food Information Resource Network (EuroFIR) (FP6‐513944).
Effects of folic acid intake on plasma folate, vitamin B12 and homocysteine levels, hepatic SAM/SAH ratio and the expression of cerebral myelin basic protein in pregnant and lactating rats
Authors: Oh Kyung Hee , Lee Ji Hye , Kim Ki Nam , Chang Namsoo
Published: June 2021

The effect of folate nutrition during gestation on the development of offspring's brain has been extensively studied, but that on the maternal brain remains mostly unknown. To investigate this, female Sprague‐Dawley rats were fed either folic acid supplemented (FS)‐ or deficient (FD)‐diet (8mg vs. 0mg folic acid/kg diet) from 2 weeks prior to the mating throughout the pregnancy and lactation period. At d20 of pregnancy and d21 of lactation, plasma folate levels and hepatic SAM/SAH ratios of the FD group were significantly lower than those of the FS group (p<0.001) with a concomitant change in the plasma homocysteine level (p<0.001). Plasma vitamin B12 level of the FD group was significantly lower than that of the FS group at d21 of lactation (p<0.01). At d20 of pregnancy, the MBP expression in the spinal cord, but not in cerebral cortex or hippocampus, was higher than that of pre‐pregnancy in all groups with a greater increase occurring in the FS group than the FD group. Plasma folate and vitamin B12 level was correlated with hepatic SAM/SAH ratio positively and with homocysteine negatively. The expression of MBP in the spinal cord, but not in cerebral cortex or hippocampus, was positively correlated with hepatic SAM/SAH ratio. These results indicate that the expression of MBP in the spinal cord of maternal rats is influenced by the hepatic SAM/SAH ratio and indirectly by dietary folic acid intake.
Protein quality evaluation of meat and fish substrates using ileally cannulated dogs
Authors: Faber Trevor A , Hernot David C. , Parsons Carl M. , Swanson Kelly S. , Smiley Scott , Bechtel Peter J. , Fahey George C.
Published: June 2021

Three meat substrates (beef loin, pork loin, and chicken breast) and two fish substrates (pollock fillet and salmon fillet) were evaluated for protein quality and amino acid bioavailability using the ileal cannulated dog assay. Pollock contained the highest amount of protein, total essential amino acids (TEAA), and total non‐essential amino acids (TNEAA) (dry matter basis) at 96.9, 43.5, and 45.3%, respectively. Salmon contained the second highest amounts at 92.8, 40.5, and 40.5% followed by chicken (90.3, 40.5, 38.8%). Beef loin had the lowest protein content (82.7%), but higher TEAA and TNEAA (38.0, 37.9%) than pork (86.2, 37.6, 37.2%). Dogs assigned to a 5x5 Latin square design were fed five different diets with each substrate as the sole protein source. No significant differences were found in ileal digestibility of protein. Values ranged from 88.9% for chicken to 90.5% for pork loin and pollock fillet. Ileal TEAA and TNEAA digestibility values were not different among sources with values between 92.3‐93.2% and 88.1‐89.8%, respectively. Total tract protein apparent digestibility values ranged from 94.4 to 94.8%, with no differences noted among treatments. These data suggest that, both in the small and large intestines of the dog, there is no difference in crude protein, TEAA, and TNEAA digestion between high quality meat and fish substrates, or within meat and fish substrates themselves.

Research support; USDA/CSREES through the University of Alaska, Fairbanks
The use of pyrosequencing to identify diet‐induced changes in the canine gut microbiome
Authors: Middelbos Ingmar S , Swanson Kelly S , Qu Ani , White Bryan A , Fahey George C
Published: June 2021

The influence of the gut microbiome on host health is becoming increasingly recognized. Although our knowledge of mammalian gut microbiomes is limited, new molecular techniques provide great opportunities. We used high throughput DNA 454‐pyrosequencing of amplicons from the hypervariable V3 region of the 16S rRNA gene to characterize gut microbiome composition in dogs fed two different diets. Six dogs were fed diets containing 0% (control; C) or 7.5% beet pulp (BP) in a crossover design with 14‐d periods. Estimates of individual microbiome diversity ranged from 198 to 513 species (cutoff: 96% similarity), slightly higher than that of humans. Three dominant bacterial phyla were detected, including Fusobacteria, Firmicutes, and Bacteroidetes. Fusobacteria decreased (P < 0.05) whereas Firmicutes increased (P < 0.05) when dogs were fed BP. UniFrac principal component analysis showed that microbiomes clustered by diet and that compositional changes in the phylum Firmicutes appeared to be a major contributing factor. These data suggest co‐dominance of three bacterial phyla in the dog gut compared to two in most mammals and that diet significantly altered gut microbiome composition.
High dietary arginine levels increase arginine utilization by thymocytes and peripheral blood mononuclear cells in broiler chicks
Authors: D'Amato Jennifer L , Humphrey Brooke D
Published: June 2021

Arginine (ARG) is utilized by immune cells for protein and nitric oxide synthesis and has a regulatory role in T cell development. In aves, ARG cannot be synthesized de novo, so immune cells are dependent upon acquiring dietary ARG. Therefore, the effect of dietary ARG levels on ARG utilization by thymocytes and peripheral blood mononuclear cells (PBMC) was determined in broiler chicks. Chicks were fed diets containing low (1.2%) or high (1.35%) dietary ARG. On d 3, 7, 14 and 21 posthatch, ARG transporter mRNA abundance was measured in thymocytes and PBMC by quantitative real‐time PCR. Thymocyte CAT‐1/3 and y+LAT1 mRNA abundance decreased from d3 to d21 (P<0.05). Thymocyte y+LAT2 mRNA abundance was dependent upon the dietary ARG level (P<0.05). Between d3 and d14, 1.2% ARG increased thymocyte y+LAT2 mRNA abundance 2.5‐fold (P<0.05) while 1.35% ARG did not change. PBMC CAT‐3, y+LAT1, and ATB0,+ mRNA abundance decreased from d7 to d21 (P<0.05). PBMC CAT‐1 mRNA abundance was dependent upon the dietary ARG level (P<0.05). On d7, PBMC CAT‐1 mRNA abundance was 2‐fold higher in chicks fed 1.35% ARG than in those fed 1.2% ARG (P<0.05). Thymocytes reduce the abundance of ARG exporter y+LAT2 mRNA and PBMC increase the abundance of ARG importer CAT‐1 mRNA when fed high ARG diets (1.35%). These results suggest that high dietary ARG levels (1.35%) increase ARG availability for immunity. Funds: California Agricultural Research Initiative.
Fructooligosaccharide alters microbiota, T‐lymphocyte subpopulations, and cytokines within gut‐associated lymphoid tissue (GALT) of BALB/c mice
Authors: Nieves Carmelo , Herrlinger‐Garcia Kelli A , Byard Alan R , Voehl Amanda K , Nguyen Christina M , Ukhanova Maria , Mai Volker , Langkamp‐Henken Bobbi
Published: June 2021

The purpose of this study was to examine the effect of the prebiotic, fructooligosaccharide (FOS), in maintaining intestinal health. Female (10 wk) BALB/c mice were fed 0% (control group), 4% or 8% FOS for 4 wk (n=9/diet) and GALT was examined. There were no additive effects between 4% and 8% FOS so data were pooled (FOS group). Intestinal microbiota composition was determined by 16S rRNA based profiling (DGGE). The cecal and fecal microbiota profiles differed between control and FOS. Mesenteric lymph node (MLN) lymphocytes were obtained, stimulated in culture with a T‐cell mitogen, PHA, and cytokine production was measured. Culture medium from FOS mice contained significantly more IL‐5 (37+6 vs 19+4 pg/ml). The percentage of MLN, Peyer's patch, and colon lamina propria and intraepithelial lymphocytes that were CD4+, CD8+, CD25+, and/or Foxp3+ was examined. The percentage of colon CD4+ intraepithelial lymphocytes was significantly lower in FOS (10.8+1.3%) vs control (17.6+2.8%). No other changes in GALT T‐cell subpopulations were noted. In conclusion, the addition of FOS to the diet changed intestinal microbiota, decreased the colonic intraepithelial CD4+ T‐cell population, and increased production of the T‐helper 2 cytokine, IL‐5, in a dose‐independent manner. Support: General Mills Bell Institute of Health and Nutrition.
Dietary blueberry attenuates obesity‐associated insulin resistance by reducing adipocyte death and macrophage pro‐inflammatory activation in adipose tissue of high fat‐fed mice
Authors: DeFuria Jason , Bennett Grace , Strissel Katherine J , Perfield James W , Greenberg Andrew S , Obin Martin S
Published: June 2021

Adipose tissue (AT) inflammation promotes insulin resistance (IR) and other obesity complications. AT inflammation and IR are associated with AT oxidative stress, adipocyte death, and the scavenging of dead adipocytes by M1‐polarized CD11c+ AT macrophages (ATMs). We test the hypothesis that blueberry (BB) supplementation of an obesigenic diet protects against these AT events. Male C57B/6j mice were fed one of three diets for 8 weeks: 10% fat (LF), 60% fat (HF) or HF containing 4% (w:w) BB powder (1:1 V. ashei and V. corymbosum). We assessed energy intake, metabolic rate, insulin resistance (insulin tolerance tests), body and AT weights, adipocyte size, adipocyte death, as well as AT gene expression by Q‐PCR of CD11c, inflammatory mediators and markers of oxidative stress (GPx3). Relative to the HF cohort, mice fed the HFB diet were significantly protected from insulin resistance. This protection was associated with enhanced antioxidant (GPx3) gene expression, reduced adipocyte death, attenuated expression of CD11c, and abrogation of ATM expression of TNF‐α and iNOS. BB supplementation had no effect on HF diet‐associated alterations in energy intake, metabolic rate, body / AT weight, adipocyte size or markers of alternative (M2) ATM activation. BB's salutary effect on IR may reflect beneficial actions of BB anthocyanins on antioxidant and inflammatory gene expression in adipocytes and ATM.

Grant Funding Source

NIH (1RO1DKO74979) and the Highbush Blueberry Council
A mechanism underlying the anti‐inflammatory action of piceatannol
Authors: Cho Han Jin , Kim Jin‐Kyung , So Hong‐Seob , Park Jung Han Yoon
Published: June 2021

Piceatannol (3,4,3',5′‐tetrahydroxy‐trans‐stilbene) is a derivative of resveratrol with a variety of biological activities including anti‐inflammatory, anti‐proliferative, and anti‐cancer activity. We assessed the mechanisms by which piceatannol inhibits inflammatory responses using lipopolysaccharide (LPS)‐treated Raw264.7 murine macrophages. Piceatannol (0 ‐ 10 μmol/L) decreased LPS‐induced release of nitric oxide, which was accompanied by decreases in the expression of the iNOS protein. The activations of NF‐κB, AP‐1, and STAT3 are crucial steps in the transcriptional activation of the iNOS. Piceatannol prevented LPS‐induced degradation of inhibitor of κBα, translocation of p65 to the nucleus, and phosphorylation of SAPK/JNK. Also, piceatannol inhibited the phosphorylation of STAT3 and translocation of STAT3 to the nucleus. In addition, piceatannol increased the protein and mRNA levels of hemeoxygenase (HO)‐1, the rate‐limiting enzyme of heme catabolism which plays a critical role in mediating antioxidant and anti‐inflammatory effects. Piceatannol inhibited ARE‐driven luciferase activity in Raw cells transfected with an ARE‐luciferase reporter construct containing the enhancer 2 and minimal promoter region of HO‐1. These results suggest that piceatannol exerts anti‐inflammatory effects via the downregulation of iNOS expression and uregulation of HO‐1 expression.