bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of OGT interacting proteins at M phase
Authors: Chambers Melody S.D. , Villar Maria T. , Artigues Antonio , Slawson Chad
Published: June 2021

O‐GlcNAc transferase (OGT) modifies numerous proteins with O‐GlcNAc, a post‐translational modification consisting of the attachment of a single N‐acetylglucosamine residue to serine or threonine amino acids on nuclear and cytoplasmic proteins. During mitosis, OGT localizes at the mitotic spindle and midbody modifying over a hundred mitotic substrates. Interestingly, how OGT is able to target discrete mitotic substrates at specific times is largely unknown. We decided to address this issue by isolating mitotic OGT interacting proteins. First, we grew HeLa cells in SILAC (Stable Isotope Labeling of Amino Acids in Cell) media and synchronized the cells either in prophase, metaphase‐anaphase, or telophase. These lysates were purified for OGT interacting proteins by binding them to an OGT column. Interestingly, silver stains of proteins eluted from the OGT columns demonstrated an increase in binding in the telophase fraction. Furthermore, this pattern was evident in OGT co‐immunoprecipitation experiments from these same lysates. These data suggest that numerous proteins form interactions with OGT at the midbody. Next, the eluted proteins were combined, digested, and the resulting peptides were identified and quantified by mass spectrometry. We found numerous proteins that interact with OGT during M phase. These results suggest that OGT is targeted to mitotic structures by discrete proteins.
Transgene regulation in plants by alternative splicing of a suicide exon
Authors: Hammond Ming Chen , Hickey Scott F , Sridhar Malathy , Westermann Alexander , Qin Qian , Vijayendra Pooja , Liou Geoffrey
Published: June 2021

Compared to transcriptional activation, other mechanisms of gene regulation have not been widely exploited for the control of transgenes. One barrier to the general use and application of alternative splicing is that splicing‐regulated transgenes have not been shown to be reliably and simply designed. Here we demonstrate that a cassette bearing a suicide exon can be inserted into a variety of open reading frames (ORFs), generating transgenes whose expression is activated by exon skipping in response to a specific protein inducer. The surprisingly minimal sequence requirements for the maintenance of splicing fidelity and regulation indicate that this splicing cassette can be used to regulate any ORF containing one of the amino acids Glu, Gln, or Lys. Furthermore, a single copy of the splicing cassette was optimized by rational design to confer robust gene activation with no background expression in plants. Thus, conditional splicing has the potential to be generally useful for transgene regulation.
Antibiotic resistance in bacteria: Structure of a novel ss‐DNA metalloenzyme inhibitor
Authors: Pyle Ronald , Fuson Kerry , Sutton R. Bryan , Morales Jorge A. , Shaw Robert W.
Published: June 2021

β‐Lactam antibiotics are among the most important drugs used to fight bacterial infection. Overuse and misuse of β‐lactam antibiotics has caused the evolution of resistance mechanisms, allowing pathogenic bacteria to survive antibiotic treatment. The major source of resistance to β‐lactam antibiotics occurs through production of enzymes called β‐lactamases capable of catalyzing hydrolysis of the β‐lactam rings in these drug compounds. The metallo‐β‐lactamases have become a major threat due to their broad substrate specificities; there are no clinically useful inhibitors for these metalloenzymes. Previously, we reported the discovery of a 10‐residue single‐stranded DNA (ss‐DNA) aptamer that is a potent inhibitor of the B. cereus 5/B/6 metallo‐β‐lactamase [S.‐K. Kim, C. L. Sims, S. E. Wozniak, S. H. Drude, D. Whitson and R. W. Shaw, Chemical Biology and Drug Design, 74, 343–348 (2009)]. The ss‐DNA may have a stem‐loop type of secondary structure. We are utilizing molecular mechanics and molecular dynamics calculations to attempt to predict the solution structure of this 10‐residue ss‐DNA aptamer. Furthermore, we have successfully crystallized this aptamer and have shown that the crystals can diffract to 2.5 Å resolution, or better. (TTU‐OTC)
Identification of Potent inhibitors of Bacillus anthracis FtsZ: A target for antimicrobial agents
Authors: Yoon Moon-Young , Park Hae-Chul
Published: June 2021

FtsZ plays an essential role in cell division and identified as antimicrobial target. The expression plasmid carrying Bantx‐FtsZ gene was expressed in Escherichia coli BL21 (DE3) cells and the expressed recombinant Bantx‐FtsZ was purified under the soluble conditions using Ni2+ charged sepharose. The purified recombinant Bantx‐FtsZ yield was estimated to be 3.5 mg/L culture. Guanosine 5‐triphosphate (GTP) dependent polymerizations of recombinant Bantx‐FtsZ were confirmed with both turbidity at A350 and Transmission Electronic Microscope (TEM). The polymerized Bantx‐FtsZ structure was confirmed with TEM that was filamentous shaped. The minimum‐required and saturation concentrations for polymerization of Bantx‐FtsZ were found to be 0.012 and 0.2 mg/mL, respectively. Berberine, a known E. coli FtsZ polymerization inhibitor, inhibited the Bantx‐FtsZ polymerization with an IC50 value of 60 ± 4.4 μM. In addition, our efforts aimed to identify potent inhibitors of Bantx‐FtsZ polymerization from a naturally extracted chemical library yielded 3 chemicals with IC50 values ranges from 10 to 100 μM. Understanding the biochemical basis and growth inhibition of B.anthracis of these inhibitors in future is essential for the identification of potential new anti‐anthracis drugs.
A New Approach for the Identification of Allosteric Binding Sites in Proteins
Authors: Striebeck Petra , Manstein Dietmar J.
Published: June 2021

Recent results indicate that small‐molecule‐protein interactions modulate the function of a large fraction of the enzyme complement in eukaryotic cells. In particular, modulation of enzyme functions by small metabolites that are not their substrates or products is increasingly recognized as a mechanism by which cells can adapt to changes in environmental conditions. While the effect of metabolite‐binding on conformational transitions and thus protein folding, stability, and catalytic activity is clearly of considerable importance, studying metabolite‐protein interactions is in most instances impeded by the low‐affinity nature of the interactions, their moderate effect on enzyme activity, and the complex nature of their interference with substrate turnover. Here, we describe an approach for the efficient mapping of the binding sites of nutrient polyphenols and small dye molecules to the 80 kDa myosin motor domain. We speculate that their binding occurs preferentially with sites that are important for protein‐metabolite interactions and can also be exploited as binding sites for therapeutic drugs. To determine the affinity of the prolific binders for the myosin‐2 motor domain and to select the most suitable compounds, we performed microscale thermophoresis experiments. Subsequently, we determined the structure of the binding pockets for the most promising compounds by X‐ray crystallography.
A novel anti‐aging compound extends longevity by remodeling neutral lipid metabolism
Authors: Titorenko Vladimir , Beach Adam , Richard Vincent , Burstein Michelle
Published: June 2021

We use yeast as a model to understand a mechanism linking longevity and lipid metabolism. Caloric restriction (CR) extends lifespan across species. We examined 1) the effect of CR and numerous mutations extending yeast lifespan on the spatiotemporal dynamics of the proteomes and lipidomes of organelles involved in neutral lipid metabolism, including the endoplasmic reticulum (ER), peroxisomes, and lipid droplets; and 2) the chronology of hallmark events of apoptotic and necrotic cell death pathways in various long‐lived yeast mutants impaired in neutral lipid metabolism. Our findings revealed that a calorie‐rich diet suppresses peroxisomal oxidation of fatty acids (FFA) that originate from neutral lipids synthesized in the ER and deposited within lipid droplets. The resulting age‐related accumulation of arrays of FFA within lipid droplets triggers a programmed necrotic cell death pathway. We designed a chemical genetic screen for small molecules that extend yeast longevity by altering neutral lipid metabolism. Our screen identified lithocholic acid (LCA) as one of such novel anti‐aging compounds. Using a combination of proteomic, lipidomic, metabolomic, and systems biological approaches, we established the mechanism underlying its life‐extending ability. We found that this bile acid remodels neutral lipid metabolism in the ER, lipid droplets, and peroxisomes, thereby postponing age‐related necrosis caused by the buildup of FFA. Our findings revealed two different ways for delaying aging by altering neutral lipid metabolism and thereby impairing the age‐related programmed necrotic death pathway. Supported by NSERC of Canada.
Adaptive Exploitation of Stromal Cell Metabolism by Tumor Cells
Authors: Patel Brij , Rattigan Yanique , Ackerstaff Ellen , Koutcher Jason , Sukenick George , Glod John , Banerjee Debabrata
Published: June 2021

BACKGROUND/OBJECTIVE

Glycolytic tumors secrete lactate into the tumor microenvironment (TME). Glycolytic muscle cells also secrete lactate, while neighboring oxidative muscle cells use this lactate as an energy source. We propose that lactate mediates tumor‐TME metabolic coupling similar to that of muscle cells. Using in vitro‐derived activated TME cells (CAFs), we investigated the role that lactate may play in CAF‐mediated support of tumor growth.

METHODS/RESULTS

[1] We find that MDA‐MB‐231 (MDA) breast cancer cells secrete 3‐fold more lactate under hypoxia, and that lactate recruits CAF precursors towards MDAs. [2] Lactate is transported by monocarboxylate transporters (MCTs); it is taken up via MCT1 and effluxed via MCT4. MDAs have low MCT1 but high MCT4 expression, while CAFs show high MCT1 but low MCT4 expression, implying that tumor‐extruded lactate is taken up by CAFs. [3] 13C NMR spectroscopic analyses indicate that 13C‐lactate is indeed metabolized via the Krebs cycle in CAFs. This implies that CAFs oxidatively metabolize lactate. [4] MDAs co‐cultured with CAFs display greater uptake of exogenous pyruvate, and subsequent 13C‐pyruvate tracking assays indicate the presence of 13C‐labeled lactate. Collectively this suggests that tumor cells may derive sources of energy from lactate‐oxidizing components of their TME. [5] Ongoing studies investigating MCT1 blockers CHC and Leutolin indicate that inhibiting lactate transport in CAFs may inhibit tumor growth.

CONCLUSIONS

To our knowledge this is the first in vitro model system demonstrating tumor/TME metabolic coupling by which tumor cells exploit stromal cells. A better understanding of the molecular mechanisms governing metabolic cooperation within the tumor milieu will potentially identify new targets for therapeutic intervention. Grant Funding Source: NIH P50 CA86438 & NJCCR‐10‐1964‐CCR‐EO

Grant Funding Source : NJCCR‐10‐1964‐CCR‐EO
Effect of Zinc on Carbonic Anhydrase IX Activity in MDA‐MB‐231 Breast Cancer Cells
Authors: Frost Susan Cooke , Tu Chingkuang , Silverman David N.
Published: June 2021

Carbonic anhydrase IX (CAIX) is a zinc‐dependent, membrane‐bound glycoprotein that catalyzes the reversible hydration of carbon dioxide like other members of the carbonic anhydrase family: CO2 + H2O → H+ + HCO3−. CAIX is also considered a marker for hypoxic tumors (including those in breast cancers), a poor prognosticator for outcome, and associated with tumorigenesis. Based on previous studies with recombinant proteins that suggested that zinc activates CAIX independent of its role in catalysis, we sought to explore the effect of zinc on CAIX activity in a breast cancer model system, the triple negative MDA‐MB‐231 cell line. These cells express only CAIX and not other membrane‐bound CA family members. We directly analyzed CAIX activity in intact cells and membrane ghosts by using 18O exchange between CO2 and water as measured by membrane inlet mass spectrometry (MIMS). Using this system, we were unable to demonstrate any effect of zinc, over a wide range of concentrations, on hypoxia‐induced CAIX activity. Further, supplementing the growth medium with zinc did not alter either CAIX expression or activity in MDA‐MB‐231 cells. Thus, it appears that zinc is only involved in the catalytic process in a biological setting.
Tamoxifen enhances chemotherapeutic efficacy of C6‐ ceramide and increases induction of apoptosis in human colorectal cancer cells by upregulation of MAPK signaling pathway and down‐regulation of inhibitor of apoptosis protein, survivin
Authors: Morad Samy , Madigan James , Rosenberg Daniel W , Kester Mark , Shanmugavelandy Sriram S , Cabot Myles C
Published: June 2021

Resistance to chemotherapy is a critical barrier to successful treatment of colorectal cancer (CRC). Ceramide, because of its role in induction of cell death cascades in cancer cells, could be a functional therapeutic alternative. Previously we showed that P‐gp antagonists verapamil, tamoxifen, and cyclosporin A block ceramide glycosylation and synergistically enhance C6‐ceramide cytotoxicity in CRC cells; however, the mechanism underlying this response was unclear. Using tamoxifen as adjuvant to C6‐ ceramide, we show the combination is highly effective in CRC cell lines. In LoVo cells, the mix induced caspase‐dependent apoptosis (DNA fragmentation, PARP cleavage, caspase activation) dosedependently, and cell cycle arrest at both G1 and G2. At the molecular level, C6‐ceramide/tamoxifen elicited early upregulation of c‐Jun N‐terminal kinase, and induced upregulation of p53 tumor suppressor protein and downregulation of survivin. Kinetic evaluation indicated that JNK was upstream of both p53 and survivin. Research during the last decade has shown that JNK, p53, and survivin are important players in resistance to chemotherapy. Here we show for the first time, the versatile, composite effects of a new drug duo that could be an effective therapy for chemoresistant CRC and possibly other cancers, solid and hematological. Supported by NIH GM77391.
Isolation and identification of the NRO1 gene, which is responsible for recognition of fatty acid species in the yeast Saccharomyces cerevisiae
Authors: Peirce Caitlin , O'Brien Regina , Heneveld Katherine , Stukey Joseph , McDonough Virginia
Published: June 2021

When dietary unsaturated fatty acids (UFAs) are present, Saccharomyces cerevisiae cells down‐regulate expression of the gene OLE1, which encodes the enzyme Δ9 desaturase, responsible for all endogenous production of UFAs. A screen was designed to isolate mutants defective in recognition of the presence of UFAs, and subsequent transcriptional regulation of the Δ9‐desaturase. Using a reporter gene construct controlled by the OLE1 promoter, several mutants were identified and one mutant, KH2, was chosen for additional characterization. The mutation that renders KH2 incapable of molecular response to UFAs is a recessive trait due to a single mutation, we call nro1 (no regulation of OLE1). KH2 does perform normal post‐transcriptional regulation of the desaturase, indicating its effect is limited to transcriptional regulation. In order to identify the gene responsible for this phenotype, a yeast genomic library was transformed into the nro1 mutant, and plasmids that returned the mutant to wild type phenotype were chosen for further evaluation. These plasmids may contain the gene NRO1, encoding the protein responsible for the sensing of exogenous UFAs. Supported by a Walker‐Barr Summer Research Scholar Award from Hope College Biology Department
ANATOMICAL MODEL IN WAX OF THE “HEART, LUNGS, POSTERIOR VIEW”: OBSERVATION AND NUMERICAL PHOTOGRAPHY IN ROTATION
Authors: Le Floch-Prigent Patrice Pierre , CHEVALIER M.
Published: June 2021

The aim of the study was to appreciate the anatomical veracity of an anatomical wax representing the heart and the lungs from a posterior view; and to realize a numerical, photographical tri‐dimensional series of photographs. The free diffusion on the internet allowed to realize a work of memory. We used a numerical photographical series (7.2 mega‐pixels) and a rotational device made of two identical superimposed marble disks linked by a ball gearing. The upper one was graduated on its periphery. The anatomical sample was carefully laid and centered on the upper disk. The pictures were shot every 5° on more than 180°. The observation of the sample allowed the identification of the main anatomical structures (heart, mediastinum, lungs), to measure it and to observe the way of fabrication. The numerical examination (with a stop on the picture) allowed to study more in details the relationships between the different organs, the structures crossing the mediastinum and their positions in the space as well. This study enhanced the quality of this work of ceroplasty realized in the end of the 19th century and also offered the veracity of the relationships existing between the different structures of the sample. The rotational acquisition of the pictures allowed to deliver them freely to every interested public.
Simple Western analysis of NFκB signaling cascade proteins
Authors: Boge Annegret , Nguyen Uyen , Ramirez Francisco , Xu Hui , Kazakova Irina , Yang Tom , Gavin Robert
Published: June 2021

Aberrant expression in the NFκB pathway is a common occurrence in inflammatory and stress‐induced diseases such as cancer. Understanding how NFκB signaling impacts disease progression is important to the development of novel therapeutics. Western blots, while semi‐quantitative and tedious to perform are commonly used. The Simple Western™ platform, provides a solution that enables the complete automation of traditional Westerns allowing analysis of up to 96 samples in a single experiment. Simple Westerns use nanoliter volumes, enabling the characterization of precious or limited quantity samples providing reproducible and quantitative results.

Targets from the NFκB pathway, including IκB, NFκB subunits c‐ Rel, p65, p50/p105 along with loading controls were run on the Simple Western platform. In response to TNF‐α treatment, statistically significant changes of key target proteins were characterized.

The Simple Western permits quantitative high throughput analysis of signaling pathways at a level of efficiency and reproducibility not obtainable by traditional Westerns, allowing better characterization of regulatory events and key biomarker targets involved in the progression of disease states.
Study of the Compartmentalization of Type 1 Angiotensin Receptor Using Bioluminescence Resonance Energy Transfer‐based Sensors
Authors: Hunyady László , Soltész-Katona Eszter , Várnai Péter , Balla András
Published: June 2021

Initiation and termination of signaling of the type I angiotensin receptor (AT1‐R) can lead to dynamic changes in its localization in membrane microdomains.

We used YFP‐labeled fusion constructs constructs (i.e. raft‐, or non‐raft plasma membrane markers) to analyze the molecular dynamics of AT1‐R trafficking in response to stimulation in bioluminescence resonance energy transfer (BRET) measurements. Our data demonstrate that angiotensin II (AngII) stimulus changes the microdomain localization of wild type or mutant (DRY/AAY or TSTS/A) AT1‐Rs in HEK293 cells. The comparison of the trafficking of AT1‐R upon AngII stimulus to either [Sar1,Ile8]‐ AngII or [Sar1,Ile4,Ile8]‐AngII stimulus revealed different type of molecular dynamics depending on the nature of the ligand of the AT1‐R. The molecular dynamics of AT1‐R also changes upon cholesterol depletion, which suggests that the mechanism is raft‐dependent. The molecular dynamics of the AT1‐R are strikingly differ from those of 5HT‐2C and EGF receptors, which demonstrate the usefulness of the BRET based measurements in the investigation of receptor trafficking in living cell experiments. These studies should provide valuable information about the distribution and molecular dynamics of AT1‐R upon angiotensin stimulus among membrane microdomains.

Supported by Hungarian National Grant OTKA NK‐100883, TÁMOP 4.2.1 B‐09/1/KMR‐2010‐0001.
VDR mutant M1T protein has diminished protein‐protein interactions
Authors: Carastro L. Michael , Morris Mark D. , Fanconi Colleen , Ladany Lisa
Published: June 2021

The Vitamin D3 Receptor (VDR) is a steroid hormone receptor protein that forms homodimers and heterodimers. The Retinoid X Receptor‐alpha receptor (RXRa) protein is a heterodimerization binding partner for VDR. A start codon polymorphism in the VDR gene, known as M1T VDR, has been reported. This VDR gene variant has been associated with increased risk for some human cancers. The mutant M1T VDR mRNA initiates protein translation at an internal Met codon and produces M1T VDR protein lacking the first three amino acids present in full length VDR protein. The goal of this work was to detect any differences in protein‐protein interactions formed by M1T VDR with VDR, M1T VDR or RXRa. We have constructed six mammalian expression vectors that produces either untagged VDR, M1T VDR, or RXRa, as well as, N‐terminally epitope‐tagged myc‐VDR, myc‐M1T VDR or myc‐ RXRa. The experimental approached employed in this study involves transient co‐transfection of H1299 cells with combinations of untagged and myc‐tagged expression vectors, followed by co‐immunoprecipitation (Co‐IP) assays and Western blotting for VDR, M1T VDR or RXRa. Our data are consistent with at least a 50% reduction in the ability of M1T VDR to form homodimers and heterodimers. These data suggest a potential molecular etiology for increased cancer risks associated with the M1T VDR mutant gene.
Engineering Plant Bioreactors For Biofuel Applications: Evolution of A. thaliana Glycosyl Hydrolases by Template Assisted DNA Shuffling and In Vitro Recombination
Authors: Davis Leroy Keith
Published: June 2021

The central aim of this research is the exploitation of in vitro evolution of plant derived glycosyl hydrolases (GHs) for conversion of plant feedstocks to self‐contained bioreactors. The quandary of lignocellulosic biofuel production is the undesired generation of fermentative inhibitors due to weak acid pretreatment. Yet, the direct expression of recombinant GH enzymes in feed stocks has the potential to alleviate the need for acid pretreatment by programmed enzymatic deconstruction of the plant cell wall. To achieve the aforementioned, a plant mesophyll protoplast model for optimization of feedstock bioreactor construction has being engineered by in vitro evolution of A. thaliana GH cDNA and subsequent introduction of engineered GHs into N. tabacum derived mesophyll protoplasts. The introduction of engineered GH genes into protoplasts allowed for analysis of GH protein expression and activity levels in plant cells. However, the dilemma of employing DNA shuffling for evolution of genes is the laborious cloning and screening required by the process. In response, whole vector template assisted DNA shuffling and in vitro recombination technology (TADSir) has been developed. The technology allows high efficiency whole vector DNA shuffling while generating sufficient DNA concentrations for direct recombination of recombinant genes into host organism of choice.
Tumor suceptibility gene 101 modulates autophagy through regulation of LC3‐II and ATG3 level
Authors: Cheng Jiin-Tsuey , Ye Chun-Cheng
Published: June 2021

Autophagy and endosomal vesicular trafficking cooperate to regulate the turnover of intracellular macromolecule, defective organelles and signaling receptor. Tumor Susceptibility Gene 101(TSG101) is a member of endosomal sorting complex required for transport‐I, ESCRT‐I, which regulates membrane receptor recycling and vesicular trafficking. Previous reports indicated that TSG101 is required for autophagy because its deprivation led to the accumulation of autophagosome. Here, we observed the increase of TSG101 and autophagic marker proteins LC3‐II and ATG3 upon nutrient starvation. Knockdown of TSG101 in HeLa cell resulted in the elevation of LC3‐II, ATG3 and ubiquitinated protein aggregates marker protein p62. However, in neuroblastoma SH‐SY5Y cell, transfection of siRNA led to the decrease of LC3‐II and ubiquitinated protein level. These results indicated that TSG101 might be essential for autophagy and the maintenance of steady‐state level of cellular ubiquitinated proteins known to be linked with the pathologic condition of neuronal cell type. Ectopic expression of HA‐TSG101 led to the increase of LC3‐II in both cell types and the elevation of ATG3 level in HeLa cell, indicating ectopic HA‐TSG101 might facilitate the execution of autophagic flow.
Structural‐functional studies of purified transmembrane segment VI‐VII of the human isoform 1 of the Na+/H+ exchanger
Authors: Alves Claudia , Lee Brian , Sykes Brian , Fliegel Larry
Published: June 2021

The Na+/H+ exchanger isoform 1 (NHE1) is a ubiquitous pH membrane protein in mammalian cells. NHE1 regulates intracellular pH exchanging one intracellular H+ for one extracellular Na+. Though the structure of membrane proteins is highly desirable, expression remains problematic. In this study we expressed and purified transmembrane (TM) VI‐VII of the NHE1 protein. A 49 kDa Maltose‐Binding‐Protein‐TMVI‐VII fusion protein was expressed in E. coli XL1 Blue cells and purified with an Amylose resin. After cleavage with Tobacco Etch Virus, free TM VI‐VII, 6.2 kDa protein was isolated and purified by reverse phase HPLC. The identity was confirmed by Mass Spectroscopy. The structure of 15N isotopically labeled TM VI‐VII peptide was determined in dodecylphosphocholine micelles using solution‐state 2D and 3D NMR. TM VI‐VII contains a helix‐kink‐helix structure for TM VI with helices between amino acids 229–236 and 239–247. TM VII was a continuous helix from 252–274. The data suggests that the C‐terminal half of TM VI interacts with the N‐terminal half of TM VII. Future studies are further defining the helix‐helix interface between residues of TM VI and TM VII.
Low Vitamin D status is a common finding in a population of pregnant women enrolled in a nutrition intervention study in the Ukraine
Authors: Carlson Charles Rock , Uriu-Adams Janet , Chambers Christina , Yevtushok Lyubov , Zymak-Zakutnya Natalya , Wertelecki Wladimir , Keen Carl
Published: June 2021

While it is known that suboptimal maternal vitamin D status can be associated with adverse pregnancy outcomes, the frequency of low vitamin D status in many populations is a subject of debate. Our ongoing study investigates the nutrient status of alcohol‐exposed Ukrainian pregnant women and whether micronutrient supplementation can reduce the risk of Fetal Alcohol Spectrum Disorder. Blood samples and alcohol consumption data are collected at the time of enrollment (mean gestational age, 20.4 ±7.0 wk). Here we present data on the baseline vitamin D status of these women. Mean plasma 25‐hydroxyvitamin D concentrations [25(OH)D] of alcohol‐exposed (n=180) and unexposed (n=180) women were 20.6 ng/mL and 21.8 ng/mL, respectively. Across both groups, 50.6% and 32.9% were classified as vitamin D deficient [25(OH)D <20ng/mL] or insufficient [25(OH)D ≥20 ng/mL and ≤30 ng/mL], respectively. When adjusted for solar radiation by month, pre‐pregnancy BMI, gestation age at blood draw, smoking status, and age, regression analyses showed that each additional oz. of alcohol consumed per drinking day, in the 2 weeks prior to the time of enrollment, was associated with a 2.4 ng/mL decrease in 25(OH)D concentrations (P=0.03). These data suggest a very high prevalence of suboptimal vitamin D status in pregnant Ukrainian women and an adverse effect of alcohol consumption on vitamin D status.
Steps/day ability to predict anthropometric changes is not affected by its plausibility
Authors: Thomson Jessica Lee , Sample Alicia , Zoellner Jamie , Tudor-Locke Catrine , Webster Michael , Connell Carol , Yadrick Kathy
Published: June 2021

We evaluated whether treating steps/day data for implausible values (<500 or >30,000) affected the ability of these data to predict intervention‐induced anthropometric (waist circumference, body mass index, percent body fat, and fat mass) changes. Data were from 269 African American participants who submitted weekly steps/day diaries during a six‐month intervention (HUB City Steps 2009–2010). Three datasets were used: full (all values analyzed regardless of plausibility), truncated (to plausible values), and excluded (if implausible). Multivariable linear regression was used to estimate the effects of steps/day on anthropometric changes. Meaningful differences in model coefficient magnitudes were not apparent among the datasets, although larger coefficients were obtained using the excluded compared to the full and truncated datasets (range of differences: 0–0.17 between full and excluded; 0–0.15 between truncated and excluded). For example, a mean increase of 1,000 steps/day resulted in 0.34, 0.37 and 0.44 cm decreases in waist circumference using the full, truncated and excluded datasets. These results suggest that manipulating steps/day data to address implausible values has little overall affect on the ability to predict intervention‐induced anthropometric changes. Funding for this research was provided by the National Institute on Minority Health and Health Disparities.
Seasonal and annual variation in intake of traditional marine foods by a Yup'ik Eskimo population: a sequential dietary record from hair stable isotope analysis
Authors: Choy Kyungcheol , Nash Sarah , Orr Eliza , Hopkins Scarlett , O'Brien Diane
Published: June 2021

There has been considerable research and public health interest in the potentially protective effects of traditional diets on disease risk in Yup'ik Eskimos. Traditional diets include high levels of fish and marine mammal intake, which provide omega‐3 fatty acids, vitamin D, and other protective nutrients. However, self‐report methods of diet assessment do not capture seasonal patterns of intake. Here, we investigate seasonal patterns of traditional marine mammal and fish intake by measuring variations in a biomarker of marine intake, the nitrogen isotope ratio (δ15N), sequentially along hair from a subset of participants in the Center for Alaska Native Health Research (CANHR) study (n = 19). We found pronounced seasonal variations in δ15N values, which we interpret as reflecting seasonal differences in the availability of specific fish and marine mammals. A pronounced summer peak in δ15N values corresponded to increased salmon availability. Furthermore, variations occurred fairly consistently across participants. Thus, participants’ ranking with respect to traditional food intake tended to stay constant. This is particularly relevant for the CANHR study, as participation varied seasonally and participants were recruited primarily during the winter months. This work was funded by a Centers of Biomedical Research Excellence grant from the NIH National Center for Research Resources (P20 RR333065).
Spatial Analyses of Healthy Eating Index Reveal a Relation between Diet Quality and Place
Authors: Aggarwal Anju , Moudon Anne Vernez , Cook Andrea , Drewnowski Adam
Published: June 2021

The Healthy Eating Index (HEI 2005) is a measure of diet quality, based on compliance with federal recommendations and guidelines. Higher HEI values have been consistently linked with higher education and incomes. However, diet quality measures still remain to be linked to specific geographic locations or neighborhoods. The Seattle Obesity Study (SOS) was a study in spatial epidemiology that collected data on home and work locations, socioeconomic status (SES), diets and health for 1,323 adult residents’ of King County, WA using a combination of telephone surveys and mailed food frequency questionnaires. Home addresses of study respondents were geocoded. Residential property values were obtained from the King County assessor tax parcel database. Multivariate regressions examined associations between HEI scores and property‐based novel metrics of personal and neighborhood SES. Higher property values were associated with significantly higher HEI 2005 scores, even after adjusting for individual level SES and demographic characteristics. Geographic Information Systems (GIS) approaches permit new spatial research on diet quality in relation to neighborhood resources and food access. These findings are the first to link HEI values, a measure of diet quality, to features of the built environment at the individual level.

Supported by NIH grants R01 DK076608 and R21 DK 085406
Association between stunting and overweight/obesity with income inequality among Mexican preschool children
Authors: García-Guerra Armando , Quezada-Sánchez Amado David , Fernández-Gaxiola Ana Cecilia , García-Feregrino Raquel , Hernández-Cabrera Amira , Gonzalez-Herrera Oscar
Published: June 2021

We used data from the Mexican National Health and Nutrition Survey 2006 from all federal entities and defined stunting and overweight/obesity (in children 2–5 y old) according to WHO criteria. Inequality levels were defined through tertiles of the Gini index. Our models included two indicator variables for medium and high income inequality, GDP per capita and schooling for each federal entity, age and sex interactions with all covariates. The prevalence of stunting was 11.6% at the lowest inequality level, 16.6% at the medium level and 19.3% at the highest inequality level. Prevalence of stunting at the medium and high inequality levels (16.6% and 19.3%, respectively) were statistically different from the prevalence at the lowest inequality level (11.6%; P=0.014 and P=0.002 respectively), no sex differences were found. Girls’ prevalence of overweight/obesity was 29.1%, 22.6% and 27.5% at the low, medium and high inequality levels, respectively. Boys’ prevalence of overweight/obesity at the high inequality level was higher than prevalence from the low (P=0.001) and medium (P=0.005) inequality levels. Our results indicate that after controlling for income there is a positive association between stunting and income inequality and a higher prevalence of overweight/obesity at the high inequality level for Mexican boys.
Contribution of flavoured milk on dietary intakes and its association with body mass index percentile in children at high risk of overweight
Authors: Wang JiaWei , Shang Lei , Johnson-Down Louise , St-Arnaud-McKenzie Danielle , Lambert Maire , Paradis Gilles , Gray-Donald Katherine
Published: June 2021

To compare children's dietary intake and body mass index percentile (BMI%ile) between those who did and did not consume flavored milk (FM). In the Québec Adiposity and Lifestyle Investigation in Youth (QUALITY) study (2005–2008), 632 Caucasian children aged 8–10 years (with at least one obese parent) were recruited. Assessment of FM, sugar sweetened beverage (SSB) and nutrient intakes was based on three 24‐hour recalls. Height and weight were measured for calculating BMI%ile. Physical activity was scaled using 7‐day accelerometer. Time spent in the sedentary behavior was collected using standard questionnaire. The percentage of overweight children was 41.9%. FM drinkers (N = 90) consumed 132g/d FM per day and had a higher mean intake for calcium (930 vs. 837 mg; P = 0.010), Vitamin D (6.9 vs. 5.9 μg; P = 0.021) and total sugar (99 g vs. 90 g, P = 0.015) than non‐FM drinkers (N = 521). These differences remained significant after adjustment for total energy intake. In multivariate analysis controlling for physical activity, sedentary behavior and SSB intake, FM consumption was not associated with BMI%ile. FM consumption was not associated with BMI%ile despite higher sugar intake and FM provided higher levels of calcium and Vitamin D intake.

Funding: Canadian Institutes of Health Research, Heart and Stroke Foundation of Canada, Fonds de la recherche en santé du Québec and Dairy Farmers of Canada
Comparison of the efficacy of popular weight loss programs in sedentary overweight women V: perception of quality of diets
Authors: Levers K , Baetge C , Lockard B , Mardock M , Simbo S , Jung Y , Byrd M , Koozehchian M , Galvan E , Khanna D , Sanchez B , Jagim A , Oliver J , Kresta J , Horrell K , Leopold T , Cho M , Rasmussen C , Kreider R
Published: June 2021

126 sedentary women (44±12 yr, 44.8±5% body fat, 35.4±6 kg/m2) were randomized to participate in a no diet or exercise control group (C) or the Curves Complete® 90‐day Challenge (CC), Weight Watchers® Points Plus (WW), Jenny Craig® (JC), or Nutrisystem® Advance Select™ (NS) weight loss programs for 12‐wks. Self‐reported diet quality was assessed on a 0–10 point Likert scale at 0, 4, 8, and 12‐wks. Data were analyzed by MANOVA and are presented as percent changes after 12‐wks. No significant interactions were seen among groups in changes in appetite (C − 12.2±20; CC −4.0±41; WW −1.6±50; JC −14.3±27; NS −3.1±33 %, p=0.69), hunger (C −16.3±31; CC −1.2±47; WW 10.4±52; JC − 0.9±47; NS 10.6±88 %, p=0.48), satisfaction with food (C −3.8±49; CC 4.3±33; WW 0.8±26; JC −3.0±34; NS 0.7±33 %, p=0.93), feelings of fullness (C −0.3±32; CC −1.1±29; WW −5.3±21; JC − 10.7±37; NS 1.4±46 %, p=0.76), or general feelings of energy (C 19.4±67; CC 55.3±67; WW 56.8±88; JC 28.4±56; NS 38.9±59 %, p=0.23). Mean changes in diet quality were greater in the CC and JC groups compared to control (C 2.2±40; CC 86±79; WW 58±68; JC 99±158; NS 64±92 %, p=0.02). Results reveal that participants generally perceived satisfaction with the diets and that diet quality was significantly better in the CC and JC diet groups compared to a non‐diet controls. Supported by Curves International (Waco, TX)
Comparison of the efficacy of popular weight loss programs in sedentary overweight women VI: quality of life
Authors: Khanna D , Baetge C , Lockard B , Mardock M , Simbo S , Jung Y , Byrd M , Koozehchian M , Galvan E , Levers K , Sanchez B , Jagim A , Oliver J , Kresta J , Horrell K , Leopold T , Cho M , Rasmussen C , Kreider R
Published: June 2021

126 sedentary women (44±12 yr, 44.8±5% body fat, 35.4±6 kg/m2) participated in a control group (C) or the Curves Complete® 90‐day Challenge (CC), Weight Watchers® (WW), Jenny Craig® (JC), or Nutrisystem® (NS) weight loss programs for 12‐wks. Participants completed the SF‐36 quality of life inventory at 0, 4, 8, and 12‐wks. Data were analyzed by MANOVA and are presented as changes after 12‐wks. Changes in perception of general health were greater in the CC group (C −0.5±9; CC 12.2±14; WW 2.2±13; JC 6.3±13; NS 2.9±10, p=0.008) while physical functioning (C 3.3±18; CC 15.6±18; WW 7.5±10; JC 18.5±41; NS 2.8±11, p=0.01) and limitations due to physical problems (C −27.8±95; CC 47.9±67; WW 40.6±88; JC 23.1±70; NS 34.6±92, p=0.05) were higher in diet groups. No differences were observed among groups in limitations due to emotional problems (C 18.5±40; CC 12.5±37; WW 29.2±72; JC 16.7±92; NS 18.5±80, p=0.98), vitality (C 1.7±8; CC 11.0±14; WW 7.9±14; JC 4.2±16; NS 6.1±11, p=0.15), mental health (C 2.9±10; CC 6.0±10; WW 5.3±11; JC 2.5±11; NS 1.5±13, p=0.56), social functioning (C 2.8±7; CC 2.1±11; WW 0.4±7; JC −0.7±10; NS −0.9±10, p=0.37), or bodily pain (C 0.2±19; CC 1.8±17; WW 0.9±17; JC 6.1±19; NS 6.3±18, p=0.77). Results indicate that participation in the CC program promotes a greater increase in perception of general health when compared to controls and other popular diet programs. Supported by Curves International (Waco, TX)
Fermented fruit‐vegetable supplementation attenuates hyperglycemia and hyperlipidemia in high fat‐fructose fed insulin resistance rat
Authors: Chao Huan-Chun , Korivi Mallikarjuna , Tsai Jyun-Han , Tung Hsiang-Yuen , Huang Hui-Yu
Published: June 2021

The present study was undertaken to investigate the anti‐hyperglycemic, antihyperinsulinemic and anti‐hyperlipidemic properties of processed food, fruit‐vegetable ferment (FVF) in insulin resistance (IR) rats. IR was induced by feeding the high fat‐fructose fed to rats. Total thirty rats were randomly divided into three groups, and severed as control, high fat‐fructose (HFD) and HFD plus FVF (HFD+FVF) groups. FVF was orally administered to HFD+FVF group at the dose of 178.5 mg/kg bodyweight for 8‐week. Oral glucose tolerance test was performed at week‐6 and week‐8. Weekly monitored fasting blood glucose and serum insulin concentrations were progressively increased in HFD group, which confirmed the IR. However impaired glucose tolerance and elevated insulin levels were significantly (P<0.01) ameliorated by FVF along with attenuated glycated hemoglobin (HbA1c) and HOMA values. Chronic HFD‐induced dyslipidemia, which was characterized by elevated total cholesterol (TC), triglyceride (TG) and low density lipoproteins (LDL) were significantly (P<0.01) attenuated in HFD+FVF group. Furthermore, decreased adiponectin and increased leptin concentrations with HFD were also reversed by FVF treatment. Our study suggests that fermented fruit‐vegetable treatment is beneficial to overcome from the high fat‐fructose‐induced insulin resistance, a pre‐diabetic state of diabetes.
Acai Juice with Baker's Yeast Beta‐Glucan Reduces URTI Symptoms and Improves Mood State in Stressed Subjects
Authors: Talbott Shawn , Talbott Julie , Vosti Mark , Blackhurst Darin
Published: June 2021

BACKGROUND

Several studies have shown Baker's yeast Beta‐1,3/1,6 D‐glucan, extracted from Saccharomyces cerevisiae is effective in reducing the incidence of cold and flu symptoms. Other studies have shown acai berry juice to provide antioxidant benefits and reduce oxidative damage.

OBJECTIVE

This study evaluated the effect of a combination of acai berry and Baker's yeast beta‐glucan (MonaVie M‐Mun) on upper‐respiratory tract infection symptoms (URTIs) and physical/psychological well‐being in subjects with moderate levels of psychological stress.

METHODS

Healthy adults (41 ± 12y) self‐administered 4 ounces of Placebo juice (N=34) or M‐mun juice (N=45) daily for 12‐weeks. We used the Profile of Mood States (POMS) psychological survey and RAND SF‐36 Medical Outcomes Survey (MOS) to assess changes in mental/physical energy levels and overall well‐being. A quantitative health perception log was used to track URTI symptoms.

RESULTS

Subjects in the M‐mun group reported 43% fewer URTI symptoms compared to Placebo, as well as superior measures of physical & mental functioning, and overall well‐being (MOS +7%, POMS +6%).

CONCLUSIONS

These data show that daily dietary supplementation with an acai/beta‐glucan juice (M‐mun) reduces URTI symptoms and improves mood state in stressed subjects, and thus may be a useful approach for maintaining immune protection against daily stressors.
Effect of glycogen synthase kinase‐3B inhibition by apigenin on markers of proliferation, inflammation and apoptosis in pancreatic cancer
Authors: Johnson Jodee , de Mejia Elvira Gonzalez
Published: June 2021

Glycogen synthase kinase‐3β (GSK‐3β) is a therapeutic target of interest in pancreatic cancer because its inhibition suppresses NFκB activity leading to decreased expression of genes that control proliferation, inflammation and apoptosis. The objective was to investigate the effect of apigenin, a dietary flavonoid, on GSK‐3β and its downstream markers. Apigenin inhibited BxPC‐3 cell proliferation (IC50 = 23 μM, 24 h). An ELISA assay demonstrated that apigenin (25 μM) decreased GSK‐3β protein expression from 4.2 ng/mL (control) to 2.3 ng/mL. qPCR results demonstrated that apigenin (IC35 = 14 μM, 24 h) affects a variety of genes related to inflammatory cytokines and receptors (27 upregulated, 13 downregulated). CCL4 was upregulated (2.23‐fold) and TOLLIP and CCR4 were downregulated (−2.29‐ and 2.60‐fold, respectively). Flow cytometry analysis demonstrated that apigenin (10, 25, 50 μM) increased the amount of cells undergoing apoptosis in a dose‐dependent manner (8, 12 and 23%, respectively). Using a cancer pathway array, to provide more insight into the potential mechanism (s) of action, demonstrated that apigenin upregulated 30 genes and downregulated 47. For instance, IFN‐β, whose activity has been shown to suppress GSK‐3β activity, was found to be upregulated 8.9‐fold. Apigenin inhibited GSK‐3β, and lead to decreased proliferation, inflammation and apoptosis in pancreatic cancer cells.
Incidence of gluteus quartus in human cadavers
Authors: Woyski Dustin , Olinger Anthony , Wright Barth
Published: June 2021

Human gluteal musculature is described as three distinct muscles: gluteus maximus, gluteus medius and gluteus minimus. There also exists a lesser‐known fourth muscle, gluteus quartus (also referred to as gluteus scansorius.) First mentions of gluteus quartus appear in scientific literature from the early 1800's, with descriptions of chimpanzee anatomy. In an 1885 paper it was proposed the human iliofemoral band is a degenerated remnant of the gluteus quartus, later refuted by a paper published in 1900. Mention of an accessory muscle anterior to gluteus medius has variably appeared in older French and German anatomy texts. A single specimen cadaveric study in 1954 gave the first detailed description of gluteus quartus in humans. A recent study of gluteus medius and gluteus minimus found gluteus quartus in 2 of 16 specimens. The purpose of our study was to determine the prevalence of gluteus quartus in both the male and female population. Careful dissections were carried out on 45 embalmed cadaveric hips (29F, 16M). Gluteus quartus was present either as small fibers or a distinct muscular bundle. The Gluteus quartus was observed as fibers in 44% of males and 41% of females, gluteus quartus was observed as a bundle in 50% of males and 31% of females. There was an overall incidence of 80%, suggesting that gluteus quartus is more prevalent in humans than previously thought.
Excessive autophagy contributes to neuron death in cerebral ischemia
Authors: Shi Ruoyang , Zhang Lin , Zhao Ling , Weng Jiequn , Li Xinmin , Kong Jiming
Published: June 2021

Aims

To determine the extent to which autophagy contributes to neuronal death in cerebral hypoxia and ischemia.

Methods

We performed immunocytochemistry, western blot, cell viability assay, and electron microscopy to analyze autophagy activities in vitro and in vivo.

Results

In both primary cortical neurons and SH‐SY5Y cells exposed to oxygen and glucose deprivation (OGD) for 6 h (hours) and reperfusion (RP) for 24, 48 and 72 h respectively, an increase of autophagy was observed as determined by the increased ratio of LC3‐II to LC3‐I and Beclin‐1 (BECN1) expression. Using Fluoro‐Jade C and monodansylcadaverine (MDC) double‐staining, and electron microscopy we found the increment in autophagy after OGD/RP was accompanied by increased autophagic cell death, and this increased cell death was inhibited by the specific autophagy inhibitor, 3‐Methyladenine (3‐MA). The presence of large autolysosomes and numerous autophagosomes in cortical neurons were confirmed by electron microscopy. Autophagy activities were increased dramatically in the ischemic brains 3 to 7 days post‐injury from a rat model of neonatal cerebral hypoxia/ischemia as shown by increased punctate LC3 staining and Beclin‐1 expression.

Conclusion

Excessive activation of autophagy contributes to neuronal death in cerebral ischemia. This study was supported by CIHR, MHRC, MICH and Canadian Stroke Network.