bluuebunny/crossref_metadata_2025 huggingface dataset:
Mitochondria and plasma membrane permeabilization by polycationic peptides derived from Cry‐protoxins
Authors: Victor Lemeshko
Published: June 2021

Polycationic peptides hold a promise as antitumoral and antimicrobial agents. Many natural and artificially designed polycationic peptides demonstrate high activity in permeabilization of biological membranes. In this work, several peptides composed of 20 amino acid residues were designed on the bases of amino acid sequences of some Cry‐protoxins. The effects of these peptides on the rat liver mitochondria were evaluated using the measurement of mitochondrial swelling and pyridine nucleotide fluorescence, as endogenous indirect indicator of the inner membrane potential changes. To study plasma membrane permeabilization by these peptides, the rat red blood cells with normal and artificially generated high membrane potentials were used (Lemeshko VV, Arch. Biochem. Biophys., 2010). Most of the designed peptides caused mitochondria permeabilization at the concentration of 4 ug/ml or less. The effect of the peptides on the red blood cell swelling was strongly potentiated by the artificially generated membrane potential (up to −200 mV, in the presence of valinomycin). The peptides with significant mitochondria permeabilizing effects and relatively low capacity to induce hemolysis were selected for further modifications to obtain peptide properties essential for antitumoral activity. (Supported by Colciencias grant #111840820380).
A Novel Nucleotide Kinase Encoded by Gene 1.7 of Bacteriophage T7
Authors: Tran Ngoc Quang , Lee Seung‐Joo , Richardson Charles Clifton , Tabor Stanley
Published: June 2021

Gene 1.7 of bacteriophage T7 confers sensitivity of phage T7 to dideoxythymidine (ddT). We have purified the product of gene 1.7 (gp1.7). It exists in two molecular weight forms. Only the C‐terminal half of the protein is required for ddT sensitivity. We show that gp1.7 catalyzes the phosphorylation of dTMP to dTDP with dTTP or dGTP as the phosphate donor. The Km of 70 μM and Vmax of 4.3 M‐1S‐1 are similar to those found for E. coli thymidylate kinase. However, unlike the host enzyme, gp1.7 efficiently catalyzes the conversion of the chain terminating dideoxythymidylate (ddTMP) to ddTDP, thus explaining the sensitivity of phage T7 but not E. coli to exogenous ddT. Gp1.7 has no sequence homology to any known nucleotide kinase, no identifiable nucleotide‐binding motif, and its activity is not dependent on added metal ion. When coupled with nucleoside diphosphosphate kinase, dTMP is exponentially converted to dTTP.
Lipid‐protein Interactions Studies in the αC418W Mutant Suffering from Slow Channel Congenital Myasthenic Syndrome
Authors: Grajales Jose Gabriel , Grajales Gary , Baez Carlos , Zhu Haipeng , Quesada Oreste , Gomez Christopher , Lasalde Jose
Published: June 2021

Cholesterol is known to modulate membrane's biophysical properties and influence function and trafficking of membrane proteins. Cholesterol levels may regulate the movement of membrane proteins from functional to nonfunctional pools. To study this, the nicotinic acetylcholine receptor (nAChR) is used as a model due to its location on a functional pool and also, recent findings indicate that a nAChR mutation (αC418W) displayed an increase in macroscopic response upon cholesterol depletion. The αC418W is the first nAChR mutation identified as cause for a congenital myasthenic syndrome. Using various approaches, we uncovered a fraction of αC418W nAChRs located in caveolin‐positive domains, captive in a non‐functional state and susceptible to cholesterol levels. Cholesterol depletion resulted in the relocation of these receptors to a functional pool. Using a transgenic mouse model bearing the αC418W nAChR, we studied the effects of Lipitor®. After 18 days of statin treatment, the number of nAChRs and the total number of endplates in the diaphragm of the mutant were reduced. As a result, a generalized weakness of the αC418W mutant is present and in some animals an average of 60% loss of spontaneous activity. Understanding the molecular basis for the side‐effects of statins is critical to prevent the collateral contraindications and also to improve therapeutic intervention in hypercholesterolemic patients.
Effect of palmitoyl CoA on ADP‐evoked vasorelaxations in porcine isolated coronary and mesenteric arteries
Authors: Alefishat Eman , Alexander Stephen , Ralevic Vera
Published: June 2021

Palmitoyl CoA (PaCoA) is an antagonist at human recombinant P2Y1 purine receptors (Coddou et al., 2003), HEK cells, human platelets (Manolopoulos et al., 2008), and rat thoracic aorta. The present study investigated the effect of PaCoA on vasorelaxation evoked by ADP in porcine isolated mesenteric and coronary arteries.

In isometric tension recordings, ADP evoked concentration‐dependent relaxations of the pre‐contracted porcine mesenteric and coronary arteries with maximal responses of 40 ± 12 and 95 ± 7% and pEC50 values of 4.26 ± 0.33 and 3.27 ± 0.85 (n = 7–12), respectively. PaCoA (10 μM) had no significant effect on relaxations in the coronary artery but abolished relaxations in the mesenteric artery. Endothelium removal abolished ADP‐induced relaxations in the mesenteric but not coronary artery.

The preferential inhibition by PaCoA of ADP‐evoked relaxations in the mesenteric, but not coronary, artery indicates the involvement of distinct mechanisms: endothelial P2Y1 receptors mediate vasorelaxation to ADP in the mesenteric artery while in the coronary artery ADP‐induced relaxation occurs via receptors on the smooth muscle that may involve adenosine release and activation of A2A receptors (Rayment et al., 2006).

Coddou et al. (2003). FEBS Lett 536: 145–150

Manolopoulos et al. (2008). Platelets 19: 134–145

Rayment et al. (2007) FASEB J 21: 577–585

Sponsored by the Jordanian government.
Targeting soluble guanylyl cyclase in experimental Parkinsonism
Authors: West Anthony R , Park Diana J , Sammut Stephen , Sunu Elizabeth , Park Michael J , Blume‐Rice Shannon , Tseng Kuei Y
Published: June 2021

Alterations in striatal cyclic nucleotide homeostasis are apparent following loss of dopamine (DA) innervation and may contribute to pathophysiological changes observed in basal ganglia circuits in Parkinson's disease (PD). This study examined the utility of systemic administration of the soluble guanylyl cyclase (sGC) inhibitor [1H‐[1,2,4] oxadiazolo‐[4,3‐a]quinoxalin‐1‐one] (ODQ) for reversing electrophysiological, histochemical, and behavioral abnormalities observed in 6‐hydroxydopamine (6‐OHDA)‐lesioned rats. Striatal single‐units recorded in DA‐depleted rats exhibited robust spontaneous firing and burst activity which was temporally associated with the depolarizing phase of local field potentials recorded in the motor cortex. ODQ administration reversed the pathological elevations in striatal unit activity and cytochrome oxidase staining observed in the subthalamic nucleus (STN) of 6‐OHDA‐lesioned rats. Moreover, ODQ treatment transiently attenuated forelimb akinesia in 6‐OHDA‐lesioned rats exhibiting incomplete (<85%) DA cell loss. These observations suggest that cGMP signaling contributes in an important manner to the overactive excitatory transmission observed in striatopallidal neurons in the DA‐depleted striatum. Thus, sGC inhibition may represent an effective therapeutic strategy for restoring motor deficits observed in PD following incomplete DA cell loss.
Involvement of Phosphoinositide 3‐kinase‐Akt signaling pathway in sodium channel activator‐induced neurite outgrowth
Authors: Jabba Sairam V , Gerwick William H , Murray Thomas F
Published: June 2021

Calcium influx through N‐methyl D‐aspartate receptors (NMDAR) regulates signaling cascades that underlie neuronal development and synaptic plasticity. Earlier we demonstrated that the voltage‐gated sodium channel (VGSC) activators, brevetoxin and antillatoxin, increased [Na+]i, stimulated Ca2+ influx through NMDARs and enhanced NMDAR‐mediated neurite outgrowth (Joju et al., 2009, Jabba et al., 2010). Here we tested whether VGSC activator‐induced neurite outgrowth may involve NMDAR‐dependent activation of the PI3K‐Akt pathway. Veratridine (VRT), a VGSC activator, was used in this study. VRT enhanced neurite outgrowth and dendritic arborization in immature cerebrocortical neurons and displayed a biphasic concentration‐response curve. VRT produced an elevation of [Na+]i and [Ca2+]i in a tetrodotoxin‐sensitive manner. Exposure to VRT increased Akt phosphorylation in immature cerebrocortical neurons. Either the NMDAR antagonist MK‐801, PI3K inhibitor LY294002 or calmodulin‐dependent protein kinase kinase (CaMKK) inhibitor STO‐609 abolished VRT‐induced Akt phosphorylation. VRT‐induced neurite outgrowth and dendritic arborization was abrogated by LY294002. These data suggest that VGSC activator‐enhanced neuritogenisis and dendritic arborization involves elevation of [Na+]i, and [Ca2+]i, and activation of the NMDAR‐PI3K‐Akt pathway.

(NIH RO1 NS053398‐06)
Mitochondria‐specific overexpression of phospholipid hydroperoxide glutathione peroxidase (GPx4) attenuates ischemia/reperfusion (I/R) associated apoptosis
Authors: Baseler Walter Allen , Williamson Courtney L. , Dabkowski Erinne R. , Croston Tara L. , Hollander John M
Published: June 2021

A primary determinant of the extent of I/R injury in the mitochondrion is antioxidant defense capacity. Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a unique antioxidant enzyme capable of reducing peroxidized acyl groups in phospholipids, providing protection to biological membranes. The goal of this study was to evaluate the impact of mitochondria‐specific overexpression of PHGPx on apoptotic susceptibility following cardiac I/R. Transgenic mice were created in which PHGPx was overexpressed in the mitochondrion (mPHGPx). MPHGPx and control hearts were subjected to global no‐flow ischemia (20 min) followed by reperfusion (90 min). MPHGPx subsarcolemmal mitochondria (SSM) displayeda decreased propensity for undergoing apoptosis as compared to I/R controls, with no differences in interfibrillar mitochondria (IFM). Caspase‐3 and −9 activities were decreased in mPHGPx SSM relative to I/R control (P<0.05), with no differences in IFM. Mitochondrial permeability transition poreopening was decreased in mPHGPx SSM as compared to control(P<0.05), with no difference in IFM. Cyclophilin D was significantly decreased in mPHGPx SSM relative to I/R control (P<0.05), with no differences in IFM. These results indicate that mitochondria‐specific PHGPx overexpression protects SSM against I/R‐associated apoptosis.

(Supported by NIH DP2 DK083095, AHA 0815406D, AHA 0855484D, NIH T32 HL090610)
Mass spectrometry quantitation and analysis of urinary metabolites of grape seed extract polyphenols in a rat model of menopause (1025.11)
Authors: Kim Helen , Cutts John , Moore Doyle , Barnes Stephen
Published: June 2021

Differences in urinary biomolecules following ovariectomy, and the impact of dietary polyphenols (grapeseed extract [GSE]) in rodents were examined by nanoLC‐tandem mass spectrometry, and the impact of dietary polyphenols (grapeseed extract [GSE]). Urines from sham‐ovariectomy (OVX) and ovariectomized ((OVX) rats were analyzed, where both groups were given GSE, thus, any differences were due to the OVX. In positive mode LC‐MS/MS, 5,589 peaks were resolved with unique retention times. Of these, 12% were significantly different in abundance (p<0.05), with 2% of this subgroup (p<0.001). Conclusive identification of significant peaks required both the METLIN software on the Scripps XCMS server (Tautenhahn et al., Anal. Chem, 2012) and the Human Metabolome Database (Wishart et al., Nucleic Acids Res, 2007). Experiments are ongoing to continue identification of peaks using MS/MS, as well as to populate the appropriate databases with conjugated forms of polyphenols to enable deeper analysis. In TARGETED metabolomics analysis, we determined that glucuronidated grapeseed polyphenols were increased in the urines of OVX rats, indicating that OVX increased the amount or activity of the relevant glucuronosyltransferases. This is the first metabolomics analysis of urinary biomolecules in a model of menopause, and of effects and fate of dietary polyphenols.

Grant Funding Source: NIH/NCCAM R21‐AT00480531
Teduglutide for safe reduction of parenteral nutrition requirements in adults: a systematic review (1035.11)
Authors: Naberhuis Jane , Tappenden Kelly
Published: June 2021

Teduglutide, a recombinant analogue of human glucagon‐like peptide‐2, is the first long‐term medical therapy approved for the treatment of adults dependent on parenteral nutrition (PN). The aim of this systematic review was to assess the safety and efficacy of teduglutide in reducing parenteral nutrient or fluid requirements in PN‐dependent adults. Studies were identified using predefined search criteria and multiple databases, including Medline and Embase. The search was completed to October 13, 2013 in the absence of date or study design restrictions. Hits were assessed for inclusion criteria and methodological quality by two independent reviewers using risk of bias assessment tables adapted from the Cochrane Collaboration. The outcomes of interest were changes in parenteral nutrient or fluid requirements (weekly volume and/or infusion days) and adverse event data. From 1,214 unique hits, 58 abstracts were reviewed. Thirty‐five reports met the inclusion criteria, including data from two phase III, placebo‐controlled clinical trials and their respective extension studies. Data provided will assess the evidence base for provision of teduglutide in adult patients dependent on PN.
Image cytometry for 2D and 3D quantification in microscopic images (1050.5)
Authors: Van Noorden Cornelis
Published: June 2021

Image cytometry is a powerful quantitative tool based on microscopy and digital imaging to obtain measurements in cells and tissues, either as stereological data (volumes, areas, lengths, profiles, etc.) or photometric data (absorbance, fluorescence and luminescence). Cytometry relies on careful calibration and control of acquisition of digital images that are evaluated with image analysis programs. The software used for cytometry may either take the form of general image analysis software or may be designed for a specific purpose. In either case, it has to be capable of assisting in the fundamental requirements of cytometry: calibration, interactivity, speed and sampling to obtain correct measurements. Stringent data acquisition and calculations are essential to obtain meaningful data. The major aim of the presentation is to provide simple but reliable procedures for valid image cytometry to obtain meaningful quantitative data from images of microscopic objects. Image cytometry instrumentation, software, validation and calibration and operation routines are discussed and protocols, hints and tips are presented.

Grant Funding Source: None
Iron chelating agents lead to the formation of neutrophil extracellular traps and subsequent entrapment of Staphylocccus aureus (1056.8)
Authors: Völlger Lena , Akong Kathryn , Dewitz Carola , Wang Yanming , Naim Hassan , Nizet Victor , von Köckritz‐Blickwede Maren
Published: June 2021

Neutrophil extracellular traps (NETs), decondensed chromatin structures that contain antimicrobial components and mediate the entrapment of microbes, have been described as a fundamental innate immune defense mechanism, in addition to phagocytosis and degranulation of neutrophils. In this study, we investigated the effect of iron‐chelating agents on NET‐formation. Primary neutrophils were isolated from human blood samples and treated with deferoxamine (DFO), AKB‐4924, or L‐mimosine. NET‐formation was subsequently visualized and quantified by immunofluorescence microscopy.

All tested iron chelators significantly induced NET‐formation and Staphylococcus aureus entrapment. Addition of iron completely abolished the induction of NET‐formation. To gain insight into the underlying mechanism, cells were treated with DPI to block NADPH‐dependent reactive oxygen species generation, which significantly abrogated the DFO‐induced NET‐formation. Furthermore, pharmacological inhibition of peptidyl‐arginine‐deiminase 4 (PAD4) resulted in significantly reduced DFO‐mediated NET‐induction. In conclusion, we demonstrate that iron‐chelation contributes to the formation of NETs. This is of particular interest because iron‐chelators used for the treatment of iron overload have become promising therapeutic candidates for fighting microbial infections.

Grant Funding Source: Supported by Deutsche Forschungsgemeinschaft
Short‐term Toll‐like receptor 9 stimulation increases blood pressure and vascular dysfunction via attenuated NO and exacerbated ROS generation (1140.9)
Authors: McCarthy Cameron , Goulopoulou Styliani , Wenceslau Camilla , Matsumoto Takayuki , Webb R. Clinton
Published: June 2021

Toll‐like receptor (TLR) 9 is a pattern recognition receptor of the innate immune system. We have previously shown that TLR 9 activation attenuates nitric oxide (NO) production in the aorta; however, the mechanism for this decrease was not determined. We hypothesized that in vivo treatment with a TLR 9 agonist (ODN 2395; three i.p. injections‐0.1 μg/i.p.) would augment aortic contractile responses via increased reactive oxygen species (ROS) generation. In adult Sprague Dawley rats, blood pressure was measured pre‐ and post‐treatment via tail cuff and concentration response curves to norepinephrine (NE; 10‐11‐10‐5 M) were performed using the myograph, in the presence or absence of inhibitors for NO synthase (L‐NNA, 10‐4 M) and ROS (tempol, 10‐3 M and apocynin, 10‐4 M). ODN 2395 treatment increased systolic blood pressure (Δ SBP Veh: ‐3±1 vs. ODN 2395: 9±3 mmHg; *p<0.05). ODN 2395 amplified the contractile response to NE [Emax (% max KCl); Veh: 108±2 vs. ODN 2395: 124±3; *p<0.05] and this difference was abolished in the presence of L‐NNA (Veh: 131±5 vs. ODN 2395: 134±2). Tempol (Veh: 116±2 vs. ODN 2395: 115±3) and apocynin (Veh: 116±2 vs. ODN 2395: 110±4) also abolished the heightened contractile response to NE. These data indicate that ODN 2395 treated animals have exacerbated ROS generation that possibly quenches NO leading to vascular dysfunction and increased blood pressure responses.

Grant Funding Source: Supported by AHA, CNPq, & NIH
Medical educators response to a web‐based nutrition implementation toolkit (WNCIT) for entry level medical courses (118.2)
Authors: Nowson Caryl , Perlstein Robyn , Beck Eleanor , Lindley Jennifer , Schafer Jennifer , Rooney Kim
Published: June 2021

Medical educators response to a web‐based nutrition implementation toolkit (WNCIT) for entry level medical courses
Caryl A Nowson1, Robyn Perlstein1, Eleanor Beck2, Jennifer Lindley3, Jennifer Schafer4, Kim Rooney5
1 School of Exercise and Nutritional Sciences, Deakin University, Australia 2 DAA, Australia & University of Wollongong, Australia.

3 Faculty of Medicine, Nursing & Health Sciences, Monash University, Australia4 School of Medicine, University of Queensland, Australia 5 School of Medicine, University of Tasmania, Australia In Australian medical courses there is no clear integration of nutrition knowledge and skills as a core component of curriculum. Four universities and the Dietitians Association of Australia (DAA) have partnered to develop a Web‐based Nutrition Competency Implementation Toolkit (WNCIT) for entry‐level medical courses. This is a systematic approach that facilitates mapping of curriculum to integrate nutrition competencies. To assess the expert views of medical educators on the development of a WNCIT, an online survey and paper‐based survey was developed. To date 37 educators have completed the survey and initial results indicate that although most respondents thought the WNCIT would be useful, they were less certain that their institution would actually use WNCIT. Barriers to implementation cited were: the already full nature of the curriculum (75%); inability to train educators and use the technology (17%); associated costs(19%). Factors that support the development and implementation of the toolkit cited were: highly relevant content (19%), ease of use(8%), clear exemplars(5%), distribution of content over whole curriculum(10%0, low cost(5%), support of higher management(10%). The results indicate that it will be important to address these issues to facilitate the widespread adoption of WNCIT in Australian medical courses.
Analysis of state of early year children nutrition status in children's home of Kazakhstan (1185.3)
Authors: Turgambayeva A. , Syzdykova A. , Tulemisova A. , Omarkulov B. , Alihanova K.
Published: June 2021

Today social orphanage is one of acute problems for our society.There are about 35 thousand social orphans at living parentsin Kazakhstan. Study results of children‐orphans as study group (n=226) and children, brought up in families (n=100) composing control group were used in study.

18,9% of children‐orphans in the age of 6‐12 months suffer from chronic malnutrition (‐2СО); 6,8% of them have growth inhibition(‐3СО); and 9,3% of them exceed measurement data of their age category,from 74 children‐orphans.

In control group 10% have growth inhibition, and 2,04% are accelerates.

From 92 studied children,in children of 12‐24 monthly ages46, 7% of study group are short ones and have growth inhibition that characterizes chronic malnutritionavailability. In 25% of them growth inhibition was revealed.

In control group 35% of children of indicated age category have chronic malnutrition.

On study results of 52 children in the age of 24‐36 months,in 38,2% of them chronic malnutrition was revealed, in 32,7% of them had growth inhibition. In control group only 18,7% of them hadchronic malnutrition, 12,7% of them had growth inhibition.

Thus, it is necessary to draw conclusion from all above‐stated, that 35,3% of children arefoster childrenof children's home of health care system considered to be short and chronic malnutritionof one or another degree, that in 2,3 times exceeds these indexes in children growing with their parents.
Unique two conjugation systems required for autophagy (237.1)
Authors: Ohsumi Yoshinori
Published: June 2021

In autophagy two ubiqutination‐like conjugation systems play essential roles in the membrane formation of autophagosome. In yeast among 18 Atg proteins essential for autophagy 8 proteins are involved in these reactions. Atg12 forms a conjugate with Atg5 via E1 (Atg7) and E2 (Atg10) enzymes. Another UBL, Atg8 is activated by the same E1 and transferred to E2 (Atg3) and finally forms a conjugate with a phospholipid, phosphatidylethanolamine (PE). These systems have several unique features. Atg5 is the only target of Atg12, and Atg12‐Atg5 conjugate is constitutively formed, and dimerizes by binding to Atg16 dimer. Not only Atg7 is a common E1 enzyme for Atg12 and Atg8, but also two systems function closely. Atg12 system is required for Atg8‐PE formation. We showed that Atg12‐Atg5 conjugate has E3 like activity for Atg8 lipidation, enhancing Atg3 activity of the transfer reaction of Atg8 to PE. In collaboration with N. Noda (IMC) and F. Inagaki (Hokkaido Univ) we have solved crystal structures of most Atg proteins involved in the conjugation reactions. Present our knowledge on the roles of these conjugation systems in autophagy will be presented and discussed.
The relation of human milk lutein, choline, and docosahexaenoic acid content to recognition memory abilities of 6‐month‐old breastfed infants (247.4)
Authors: Cheatham Carol , Stegall Julie , Sheppard Kelly , Armer Andrea , Millsap Grace , Wernimont Susan , Zeisel Steven
Published: June 2021

OBJECTIVE: The aim of this study was to explore the relation of human milk lutein, choline, and DHA with recognition memory abilities of 6‐month‐olds.

METHODS: Two hundred breastfeeding dyads were enrolled when the infants were 3‐4 months of age. Milk samples were obtained and analyzed for fatty acids, lutein, and choline. One hundred two of the 200 were invited at age 6 months to participate in an electrophysiology session. Recognition memory was tested with a 70‐30 oddball paradigm during which high‐density 128‐lead event‐related potential (ERP) data were collected. Sixty‐seven infants provided usable ERP data; of those, 59 had complete nutrient data. The differences between latency to minimum, minimum amplitude, and mean amplitude when viewing familiar and novel stimuli were analyzed by averaging activity at 6 different collections of leads (Frontal Right, Frontal Central, Frontal Left, Central, Midline, and Parietal) and subjecting the data to stepwise regression modeling.

RESULTS: Final regression models revealed significant interactions. The lutein by free choline interaction was significant for the difference in latency scores at Frontal Left and Central areas (p<0.05 and p<0.001, respectively). Higher choline levels with higher lutein levels were related to better recognition memory. The DHA by lutein interaction was significant for the parietal area in mean amplitude and minimum amplitude (p<0.05). Recognition memory was higher when DHA was high and lutein was low.

CONCLUSION: Interactions between human milk nutrients appear important in predicting infant cognition, and there may be benefit associated with specific nutrient combinations.

Grant Funding Source: Nutrition Obesity Research Center (NIH grant), University of North Carolina at Chapel Hill, The Bill
Cognitive development among preschoolers improves in relation to endline ferritin and vitamin D levels only among recipients randomized to a fortified poultry‐based spread (Spammy™) intervention group (250.6)
Authors: Bracken Bruce , Bonorden Melissa , de Salazar Maria , Salazar Paola , Hernandez Liza , Salguero Sylvana , Armas Laura , Solomons Noel
Published: June 2021

Background: Dietary improvements during early childhood have potential to both reduce nutritional deficiencies and improve cognitive outcomes.

Objective: To assess the efficacy to improve cognitive function of a poultry‐based spread that provides between 20% to 333% of the RDA of 12 selected micronutrients, including vitamin D and iron.

Methods: Sixty preschoolers, aged 4‐7 years, were randomized in a triple‐blinded format to receive 43 g of fortified (FS) or unfortified (US) SpammyTM during weekdays over a 20 week period as part of the institutional breakfast on the outskirts of Guatemala City. The Spanish language Bracken Basic Concept Scale‐ Receptive (BBCS‐R) was administered and circulating nutrient status indicators (25(OH)vit D and ferritin) were measured at both ends of the trial.

Results: Reliability analyses for both the BBCS‐R School Readiness Component (SRC) and total test (TT) were high (.96 and .95, respectively). Both FS and US groups had larger than expected gains on the age‐standardized SRC scores (p=.068 and 0.03, respectively) with no difference between groups (p=.84). In the FS only, a significant, positive relationship was found between TT cognitive gain score and both endline vitamin D (Pearson r = .37) and ferritin (Pearson r = .42).

Conclusion: Regular consumption of a fortified poultry spread to breakfasts, combined with educational programming, set the stage for improved cognitive functioning.

Grant Funding Source: Supported by McGovern‐Dole Food for Education and Child Nutrition Program and Hormel Foods
In the shadows of the membrane: A tale about how to beat the odds in cellular copper acquisition (360.3)
Authors: Unger Vinzenz , Pope Christopher , Flores Adrian , DeFeo Christopher
Published: June 2021

Critical for cellular function, ions of the transition metal copper need to reach multiple specific destinations inside cells in an efficient manner. That they do is taken for granted. Yet, how exactly does an ion of 11Å3 find binding sites of ~1000Å3 in a volume of 30,000,000,000,000Å3 that is accessible to diffusible species within the cytosol of even a small organism like yeast? The odds for this to happen through simple 3D‐random walks and diffusion are staggeringly low, and yet, live goes on as though this problem did not exist. Studying the mechanisms of cellular copper acquisition and distribution, we recently made the discovery that cells overcome the odds associated with targeted copper delivery by reducing the dimensionality of the search problem through exploitation of cellular membranes as scaffolding components. Specifically, our studies showed that cellular copper chaperones are able to associate with membranes and that this novel property of the chaperones is important for initial copper loading into the chaperone’s copper binding site. Taken together, our findings blend into a novel paradigm in which cellular membranes play a pivotal role in the intracellular distribution of copper ions.
Dietary glycemic index does not influence adaptation of resting energy expenditure during weight loss (371.7)
Authors: Karl J. , Schaefer Ernst , Fuss Paul , Das Sai Krupa , Saltzman Edward , Roberts Susan
Published: June 2021

It has been suggested that low glycemic index (GI) diets may help prevent weight regain following weight loss by attenuating the decline in energy expenditure that is associated with lower body weight. To determine the independent impact of dietary GI on resting energy expenditure (REE) during and after weight loss, 79 overweight and obese adults (mean ± SD; 56 ± 6 y, BMI 33 ± 2 kg/m2) were randomized to 1 of 4 provided diets differing in fat proportion (15% or 30% of energy) and GI (low, 55 or high, 88), but matched for protein and fiber contents. Food was first provided at 67% of measured weight maintenance requirements for 12 wk, then at 100% of requirements for 5 wk. Body composition and measured REE (mREE) were determined at baseline, 12 wk and 17 wk. Predicted REE (pREE) at 12 wk and 17 wk was calculated from age, sex, fat mass (FM) and fat free mass (FFM) based on a regression model developed from baseline data (R2 = 0.91, p < 0.001). Measured decreases in body weight, FM, FFM and REE over 17 wk did not differ significantly by dietary fat or GI. In the combined‐cohort, mREE was 56 ± 89 kcal/d and 44 ± 108 kcal/d lower than pREE at 12 wk (p < 0.001) and 17 wk (p < 0.01), respectively, but did not differ by dietary fat or GI‐level (12 wk: p = 0.51, 17 wk: p = 0.83). Dietary GI does not appear to attenuate the reduction in REE associated with weight loss when potentially confounding factors such as dietary protein level are controlled.

Grant Funding Source: Supported by NIH grant RO1HL 57981
Identification and quantification of anthocyanins in the transgenic tomatoes (393.8)
Authors: Su Xiaoyu , Xu Jianteng , King Brenee , Lim Wansang , Katz Benjamin , Park Sunghun , Tomich John , Wang Weiqun
Published: June 2021

Anthocyanins, a sub‐class of flavonoids, are natural pigments derived from phenylpropanoid pathway. Most tomato cultivars found in nature have very low content of anthocyanins, but dark purple tomatoes by ectopic co‐expression of two transcription factors Delila (Del) and Rosea1 (Ros1) from snapdragon and chalcone isomerase (CHI) from onion accumulated high levels of anthocyanins. This study is to identify and quantitate anthocyanins in these transgenic tomato lines. Seven anthocyanins including two new anthocyanins [malvidin‐3‐(p‐coumaroyl)‐rutinoside‐5‐glucoside and malvidin‐3‐ (feruloyl) ‐rutinoside‐5‐glucoside] have been identified in transgenic lines by HPLC‐MS. The top two anthocyanins are petunidin 3‐(trans‐coumaroyl)‐ rutinoside‐ 5‐glucoside and delphinidin 3‐(trans‐coumaroyl)‐rutinoside‐5‐glucoside, that contribute for 85% of total anthocyanins in whole fruit. Comparing with undetectable anthocyanins in the wild type, Del/Ros1‐expressing tomatoes contain total anthocyanins at 4.95±0.42 g/kg dry matter in whole fruit, 5.09±0.62 g/kg dry matter in peel, and 5.56±0.29 g/kg dry matter in flesh, while CHIxDel/Ros1‐coexpressing tomatoes have 9.61±0.71 g/kg dry matter in whole fruit, 29.9±1.64 g/kg dry matter in peel, and 8.65±0.39 g/kg dry matter in flesh. No anthocyanins are detectable in the seeds of each line tested. Enrichment of tomato fruit with new and high anthocyanins may provide potential health‐promoting benefits

Grant Funding Source: USDA Cooperative KS511‐1001903
Introducing bioethics in a molecular pathophysiology course using a popular science nonfiction book (531.26)
Authors: Walton Kristen
Published: June 2021

Students in the biology department at Missouri Western State University are exposed to bioethics issues informally in several classes, but are not required to take a separate bioethics course. The Immortal Life of Henrietta Lacks by Rebecca Skloot provides a discussion of the life of the woman whose cervical cancer biopsy gave rise to the HeLa cell line, as well as a historical context for the relevant medical, social, and ethical issues. This book is assigned reading in an upper division Molecular Basis of Disease course with two aims: 1) To expose students to the fact that many human cell lines and other research reagents originated as patient tissue, and 2) to use the Lacks family story as a springboard for discussion of the guidelines for ethical treatment of human research subjects. Students had weekly reading assignments with online submission of responses to questions about the science and ethics relevant to each assignment. After finishing the book, students were assigned to write a brief paper on the events that led to the current federal regulations for human subjects research. Students were also surveyed about their knowledge of these guidelines and for their qualitative feedback to the usefulness of including this book as part of the course. Many students reported surprise at some medical procedures and practices that were standard several decades ago. Most students were unaware that human tissues are not subject to the same regulations as human subjects in clinical research studies or the events such as the Nuremburg trials that initiated the first versions of ethics guidelines. Overall, this book has been a successful platform for increasing student knowledge of human subjects research ethics and empathy for the humans who have contributed tissues to medical research.
Insights into neural crest phosphoregulation through the antiphosphatase Paladin (541.8)
Authors: Roffers‐Agarwal Julaine , Hutt Karla , Stronge Edward , Gammill Laura
Published: June 2021

The neural crest is a multipotent, migratory, embryonic cell type that forms a variety of derivatives including the craniofacial skeleton and peripheral nervous system. While the transcription factor network that regulates neural crest formation is heavily scrutinized, the mechanisms that control where resulting proteins act is relatively ignored. We propose that modulation of protein activity through phosphorylation also regulates the establishment of neural crest cell fate. Our previous work showed that precise levels of the antiphosphatase Paladin are required for normal neural crest specification and migration in chick, emphasizing the importance of phosphoregulation during neural crest development. To elucidate the mechanism of Paladin’s action, we identified putative Paladin interactors in an in vitro screen. One interactor, Myosin Heavy Chain 9 (MYH9), is enriched in premigratory and migratory neural crest cells and mutations are associated with human patients with neurocristopathies. We are currently working to confirm the Paladin‐MYH9 interaction in vivo, examine the regulation of MYH9 phosphorylation during neural crest development, and identify novel neural crest regulators through an in vivo Paladin interactor screen. Together, this work will expand our understanding of an underappreciated level of control within the neural crest developmental program.

Grant Funding Source: Supported by NIH F32DE019973; K22DE015309; R03DE023368; Amer Assn of Anatomists; MN Med Foundation
PrPC associates with a multimolecular complex including LRP1 and glycosphingolipids within lipid rafts (601.1)
Authors: Mantuano Elisabetta , Tasciotti Vincenzo , Garofalo Tina , Sorice Maurizio , Misasi Roberta , Mattei Vincenzo
Published: June 2021

The cellular form of prion protein (PrPC) is a highly conserved cell surface GPI‐anchored glycoprotein that was identified in cholesterol‐enriched, detergent‐resistant microdomains (‘rafts’). Like other GPI‐anchored proteins, most of PrPC molecules were found in lipid rafts from neural and non‐neural cells. Recently, PrPC was identified as a high‐affinity receptor for Aβ oligomers. Prion protein‐mediated toxicity of Aβ oligomers requires lipid rafts and the transmembrane low density lipoprotein receptor‐related protein‐1 (LRP1). LRP1 is an endocytic and cell‐signaling receptor for diverse ligands, which control the activity of LRP1 by directing the recruited co‐receptors. LRP1 controls both the surface and intracellular trafficking of PrPC in neurons.

Our results show that PrPC is strictly associated with gangliosides in microdomains of neuroblastoma cells. Scanning confocal analysis reveals a consistent co‐localization between PrPC and GM1, as well as between TrkA and GM1, indicating the existence of a glycosphingolipid‐enriched molecular complex. These data suggest that LRP1, PrPC and TrkA may function as part of a single system associated with lipid rafts, involved in receptor‐mediated neuritogenic pathway. In addition, we confirmed by co‐immunoprecipitation experiments a specific PrPC ‐ganglioside interaction. Moreover, signal transduction experiments on neuritogenic pathway were performed using human recombinant PrP (hPrPrec), and the results indicate a role for PrPC in cell signaling in neurons. We hypothesize that the ability of PrP to induce cell signaling might involve a multi receptor complex, in which LRP1, in association with other receptors, like TrkA receptor, plays a role in signal transduction within lipid rafts.These findings suggest that PrPC associates with a multimolecular complex, including, LRP1, TrkA and ganglioside GM1, which is dependent on the integrity of lipid raft and is involved in the neuritogenic signaling and trafficking.
Brain dynamics and attentional control as a function of body iron status (619.4)
Authors: Rhoten Stephanie , Wenger Michael , Murray‐Kolb Laura , Gahutu Jean‐Bosco , Lung’aho Mercy , Haas Jere
Published: June 2021

Iron deficiency (ID) is a highly prevalent micronutrient deficiency, affecting individuals in both developing and developed countries. Previous research on children with ID report deficits in attention related to variations in brain dynamics as measured using electroencephelography (EEG). We report here the results of analysis of baseline data from a randomized, double‐blind dietary intervention study involving 257 female students (ages 18‐26) at the University of Rwanda, Huye; 54 of these participated in the behavioral and EEG testing, using 3 tasks designed to assess attentional capture and control: (a) simple reaction time (SRT), a test of perceptual processing speed; (b) Go/No‐Go (GNG), a test of inhibitory control; and (c) the Attentional Network Task (ANT), a test of three critical functions of attention. EEG data were collected using a 64‐channel system (BrainProducts, Gilching, Germany), with signals sampled at 250 Hz. We observed statistically significant relationships between blood measures of iron status and the magnitude (positive) and latency (negative) of two EEG components (P1/N1 and P3); a positive relationship with one measure of overall brain activity (global field power) and a positive relationship with one measure of brain activity specific to engaged attention (power in theta‐band activity). These results suggest a critical role for iron in brain dynamics related to attention.

Grant Funding Source: HarvestPlus/IFPRI
Differences in cognitive flexibility between healthy weight and obese children: an ERP study (629.6)
Authors: Khan Naiman , Raine Lauren , Drollette Eric , Scudder Mark , Pontifex Matthew , Hillman Charles
Published: June 2021

Obesity is associated with cognitive decline in later adulthood; however, the implication of obesity on childhood cognitive and brain health remains controversial. Accordingly, this study investigated the effect of weight status (healthy weight [HW] or obese [OB]) on behavioral and neuroelectric measures of cognitive control among 80 prepubertal children (8‐9‐years). HW (n=40) and OB (n=40) children completed a switch (cognitive flexibility) task while event‐related brain potentials were concurrently measured. Adiposity and aerobic fitness were assessed using DXA and VO2max, respectively. Groups were matched on key covariates including age, sex, IQ, SES, fat‐free VO2max, and ADHD. There were no differences in response accuracy or reaction time between groups on both the homogenous and heterogeneous trials. However, obese children exhibited greater global switch costs related to response accuracy. In addition, obese children displayed smaller central‐parietal P3 amplitude during heterogeneous trials, signifying decreased allocation of attentional resources. Given that global switch costs reflect efficiency of maintaining task sets in working memory as well as the selection of subsequent tasks, our findings indicate that obese children may exhibit lower cognitive flexibility when faced with greater executive function demands.

Grant Funding Source: Supported by NIH HD055352
Glucose restriction, ketone supplementation, and hyperbaric oxygen therapy inhibit proliferation and viability and increase reactive oxygen species production in VM‐M3 glioma cells (644.18)
Authors: Poff Angela , D'Agostino Dominic
Published: June 2021

The Warburg Effect and tumor hypoxia underlie a unique cancer metabolism characterized by glucose dependency and mitochondrial dysfunction. Previous research indicates this phenotype can be exploited therapeutically with glucose restriction, ketone supplements, or hyperbaric oxygen (HBOT). We have previously shown that these conditions decrease tumor growth and metastatic spread and increase survival time in mice with VM‐M3 metastatic cancer. To further characterize the effects of these therapies, we measured proliferation, viability, and ROS production in VM‐M3 cells exposed to these conditions: high glucose (control; 25mM), low glucose (LG; 3mM), ketone supplementation (βHB; 5mM), hyperbaric oxygen (HBOT; 90 min, 2.5 ATA), or combination therapy (LG+BHB+HBOT). Proliferation of VM‐M3 cells was measured with trypan blue hemocytometry and was significantly decreased from controls at 24, 48, 72, and 96 hours (p<05). Viability of VM‐M3 cells was measured by fluorescence microscopy with calcein and EthD‐1 and was decreased by 19.2% in LG, 12.5% in βHB, and 38.2% in LG+βHB+HBOT treated cells compared to control (p<0.05). ROS production was measured by fluorescent microscopy with DHE and was increased by 46% in HBOT and by 48% in LG+βHB+HBOT treated cells compared to control (p<0.001). This study further supports the use of these metabolic therapies as a potential cancer treatment.

Grant Funding Source: Supported by Office of Naval Research and a charitable donation from Scivation Inc.
Effects of Perilla frutescens on modulations of CYP1A1/2, HO‐1 and activation of Nrf2 in oxidative stress‐induced hepatotoxicity (647.19)
Authors: Kang Jeong Han , Yang Sung‐Yong , Lee Kwang‐Won
Published: June 2021

We have previously reported protective effect of Perilla leaves extract (PLE) against tert‐butyl hydroperoxide (t‐BHP)‐induced liver oxidative damage in rats. t‐BHP is known that can be metabolized to free radical intermediates by Phase I enzyme, cytochrome P450s (CYPs), and we speculate that oxidative stress was induced by t‐BHP, which seemed to be closely linked with the oxidative metabolism mediated by CYPs. We examined the effect of the PLE about CYPs related to oxidative stress on enzyme assay such as 7‐ethoxyresorufin 0‐deethylase (EROD, CYP1A1), 7‐methoxyresorufin 0‐demethylase (MROD, CYP1A2), erythromycin N‐demethylase (ERDM, CYP3A), and p‐nitrophenol hydroxylase (PNPH, CYP2E1) in rat liver. The pretreatment of rats with PLE (250, 500 and 1000 mg/kg b.w.) for 5 days before a single dose of t‐BHP (i.p.; 0.5 mmol/kg). PLE was found to be a potent inhibitory effect CYP1A1/2 of oxidative stress‐induced liver. Kinetic analysis of CYP1A1/2 activities demonstrated that PLE inhibited enzyme activities by competitive or noncompetitive mechanism, respectively. Therefore, selective control of CYPs by PLE might contribute to protective effect through inhibitory CYPs activity against oxidative stress in liver. Heme oxygenase‐1 (HO‐1) is an important antioxidant phase II enzyme that plays a critical role in protection against oxidative stress. We confirmed that PLE increased induction of HO‐1 expression as well as activity in t‐BHP‐induced liver. Also, PLE resulted in enhanced nuclear translocation and antioxidant response element (ARE)‐binding of NF‐E2‐related factor 2 (Nrf2) that induce HO‐1.
Evaluation of antifungal herbs on filamentous fungi using a rapid screening technique (657.7)
Authors: Phung Brian , Ahn Aaron , Kim Hyesoo , Bennett Edwin , Fradinger Erica , Bell Priscilla
Published: June 2021

Phytochemicals have been studied as antifungal agents due to the growing resistance of fungi to current medications. The demand of these reagents has increased due to the rising in number of immune‐deficient patients worldwide. Current methods for screening antifungal herbs involve the lengthy CLSI method (M38‐A2). Here we used a novel rapid screening technique to evaluate the inhibition of filamentous fungal growth by Bayberry Root, Elecampane, Goldenrod, and Wood Betony. Filamentous fungi were grown on potato dextrose agar plates and ground in Delbecco Medium with Fetal Bovine Serum. The slurry was filtered using a #1 Whatman filter and used for testing with the various herb preparations. Herbs were extracted with methanol, filtered, rotoevaporated, and then diluted in DMSO. Initial transmittance at 625 nm was taken after preparation and subsequent readings were taken hourly after overnight incubation at 37°C. Bayberry Root, Elecampane, Goldenrod, and Wood Betony yielded inhibition percentages of 32.1 ± 3.7, 47.1 ± 7.9, 34.5 ± 15.8, and 10.0 ± 4.7 respectively, at 50% growth of the control fungal preparation. These data demonstrate that our herb preparations show significant inhibition of fungal growth and this study will be expanded to include related filamentous fungi, as well as, additional herbs.