bluuebunny/crossref_metadata_2025 huggingface dataset:
Eustress humor associated laughter compared to stress differentially modulates EEG power spectral density across frequency bins 1‐40Hz (684.10)
Authors: Berk Lee , Alphonso Clarice , Thakker Neha , Nelson Bruce
Published: June 2021

Eustress and distress are complex processes that can differentially modulate positive or negative aspects of psychoneuroimmunological (PNI) mechanisms. The eustress of humor associated mirthful laughter (HAML) relate to fundamental aspects of human behavior. Some experiences are characterized by modulating PNI mechanisms and behavior for positive health benefits. No known electroencephalographic (EEG) study has been conducted to differentiate Power Spectral Density µV2 (PSD) correlates across EEG frequency bins1‐40Hz for comparison of eustress HAML and distress. Purpose: This study assessed EEG PSD in 31 university students during HAML and distress. Methods: In a cross‐over design each subject was randomly administered two 10 minute videos: HAML (America's Funniest Home Videos) and Stress (Movie ‐ Saving Private Ryan). EEG wave band activity was recorded from 9 scalp locations F3, Fz, F4, C3, Cz, C4, P3, POz, and P4 using the EEG B‐Alert 10X System™. Results: HAML compared to distress showed Beta Band Activity (13‐30Hz) (βBA) (p=0.01) and Gamma Band Activity (γBA) (31‐40HZ) (p=0.001) for PSD greatest for all 9 regions. Most profound was HAML γBA was significantly greater than HAML βBA (p=0.01) for the same 9 regions. θBA (3‐7Hz) and αBA (8‐12Hz) for P3 vs P4 showed HAML greater PSD in the P3 region than P4 (p<0.01). Frontal region γBA and βBA was different from θBA and αBA for HAML p< .05. Central βBA and γBA, distress was different from HAML, p<.0001. Parietal αBA, βBA and γBA, distress was different from HAML (p<0.05). Conclusions: HAML shows EEG PSD correlates of change for specific bandwidths but greatest for γBA across all measured cerebral cortex regions (synchrony). Previous research show spectral density γBA is suitable for EEG‐based emotion classification of happiness and sadness. Our EEG findings further suggest eustress HAML can increase PSD of γBA synchrony and thus potentially enhance cognition, sensory perception, memory/recall, happiness & other health benefits.
Intravenous mesenchymal stem cell administration spurs endogenous hepatic mitochondrial gene expression in the diet‐induced obese mouse (693.10)
Authors: Newell Christopher , Hughey Curtis , Pihl Carin , Hittel Dustin , Shearer Jane
Published: June 2021

Intravenous human mesenchymal stem cell (MSC) administration alleviates hepatic oxidative stress in the diet‐induced obese mouse. However, the underlying mechanisms responsible for these benefits are uncertain. Possible explanations range from direct cell engraftment to paracrine‐mediated endogenous cell stimulation. To examine this question, male C57BL/J mice (n=24) were assigned to one of two dietary groups; chow (10% kcal fat) and high fat (60% kcal fat) for 12 wk. Animals were further divided, receiving intravenous saline (0.2 mL PBS) or MSC (750,000 cells). Post‐injection (24 h), qRT‐PCR was used to identify changes in endogenous (mouse) and exogenous (human) transcripts for glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) and mouse mitochondrial protein cytochrome c oxidase subunit II (COX‐II) in the liver. Human GAPDH and COX‐II were detected in the livers of animals undergoing MSC treatment indicating successful cell targeting. Declines in mouse COX‐II caused by HF were mitigated by MSC therapy (p<0.05). Results indicate that MSC administration triggers endogenous mitochondrial gene proliferation which may explain the ability of MSC to mitigate hepatic oxidative stress in diet‐induced obesity.

Grant Funding Source: Supported by MitoCanada
Students' satisfaction and burnout: cross‐sectional study at the New Medical Studies in English Program of the University of Split School of Medicine, Croatia (719.17)
Authors: Valic Zoran , Malički Mario , Bradaric Kristina , Jeličić Karlo , Bozic Josko , Marusic Matko , Ljutic Dragan , Valic Maja
Published: June 2021

Introduction: Quality of teaching and the faculty’s teaching practices can be indirectly assessed through students’ satisfaction and burnout with their studies, factors which also influence students’ academic success and the schools prestige.

Aim: To assess students’ satisfaction and burnout at the New Medical Studies in English Program (started in 2011) at the University of Split, Croatia.

Methods: In October 2013, we conducted a previously designed satisfaction survey, as well as the Oldenburg Burnout Inventory (OLBI) on all students of the New Medical Studies in English in Split.

Results: In total 88 (78%) of students returned the completed questionnaires. Students GPA (out of max 5.0) was 3.4 for the 2nd year and 3.0 for the 3rd year students. Most students (87%) rated campus safety as above average or excellent, 83% rated as such the staff friendliness, 73% the teaching quality, and 67% the overall satisfaction with the School, with no differences between the study years. Students scores on OLBI subscales were 3.0±0.4 for disengagement, and 2.7±0.7 for exhaustion, with first year students showing less exhaustion than higher year students (ANOVA, P=0.010).

Discussion: Students grades, as well as self reported study satisfaction and burnout demonstrate the balance of high quality teaching and student engagement at the pre‐clinical courses of the New Medical Studies in English at the University of Split, Croatia.

This study was supported by University of Split School of Medicine.
Assessing team‐based learning method’s effectiveness in medical pharmacology teaching (719.8)
Authors: Mehnaatamai Mohanram Arun , Zhong Qing
Published: June 2021

One of the major challenges medical school students face is the arduous task of learning enormous amount of material in a short period of time. Team Based Learning (TBL) is one among the several methods that are being increasingly used in medical schools around the world, to make the learning process easier and more effective. In the medical school in Grand Cayman, TBL is being used for over 4 years as an effective way to actively involve students in the learning process. This study aims to assess the student perceptions of TBL both in the short term and long term. In the medical pharmacology course (MPC), TBL sessions include an individual online quiz, followed by students working in groups to answer the same quiz after active discussion. In order to quantify the student perceptions of TBL, we conducted a point based survey among the students taking MPC currently and in those who completed the course 4 months earlier; to assess the short‐term and long‐term impact respectively. The survey questionnaire consisted of 20 items that can be answered in 20 minutes time. The items in the questionnaire were designed to address the student perceptions of TBL design, content and team work. The survey also had an open box to capture any additional comments by the students. The responses were quantified by way of points (Strongly disagree= ‐2, disagree= ‐1, neutral = 0, agree= +1, strongly agree=+2) and the sum of the responses obtained from the current students was compared with the responses from the past students. Both the current and the past students of MPC had positive learning experience. Although the current students had a more positive experience, majority of them said that they did not prepare for the TBLs but learnt from them. The results of this study are the key drivers for many changes we are planning to make to the TBL sessions’ frequency, design and content.
Outsourcing anatomists: a model for expanding educational outreach and providing supplemental revenue to anatomy departments (721.22)
Authors: Brokaw James , Jones Kathryn
Published: June 2021

The need for qualified anatomy instructors is rapidly expanding across the educational spectrum, from high school anatomy and physiology classes, to college pre‐professional programs, to newly established health professional schools. In response to this growing demand for anatomical expertise, the Department of Anatomy and Cell Biology of Indiana University (IU) School of Medicine established the IU Center for Anatomical Sciences Education (IU‐CASE) to leverage its ample faculty resources in the pursuit of additional teaching opportunities outside the MD curriculum, thereby expanding the scope of the department’s teaching mission to include students who may not otherwise have the chance to learn anatomy. Utilizing anatomy faculty from across the medical school’s nine‐campus system, IU‐CASE provides fee‐based educational services to other schools and programs in Indiana. Current clients include Marian University College of Osteopathic Medicine, IU School of Health and Rehabilitation Sciences, IU School of Medicine at Muncie, Purdue University Department of Biology, and Indianapolis‐area high schools. Now in its first year of operation, IU‐CASE promises to earn significant supplemental revenue for the department, which can be used to offset salary inequities, provide bonus pay for teaching‐intensive faculty, and support departmental programs.
Clinical Anatomy League of University of São Paulo Medical School Methodology: analysis of the impact on student’s learning (721.24)
Authors: Iuamoto Leandro , Nogueira Thiago , Sousa Braian , Ribeiro João , Hojaij Flávio , Akamatsu Flávia , Andrade Mauro , Jacomo Alfredo
Published: June 2021

Anatomical knowledge plays an important role in medical education and practice. Learning anatomy is frequently associated with memorization. Studies have suggested that the way Anatomy is taught at Medical School is rather inappropriate and that the ability to reason and to apply this knowledge is more important than structure names memorization. The Clinical Anatomy League of University of São Paulo Medical School represents acomplementary way of learning this subject by promoting cadaver dissection activities and lectures that aim to elucidate the clinical and anatomical correlations regarding frequent procedures in the practice of medicine.

A 20‐question examination was used to assess the anatomy knowledge of 33 FMUSP students. 14 of those students were League members and the other 19 students composed the control group. The grades of the students on the undergraduate topographical anatomy course and on this research project’s examination were used in order to compare the capability of retaining the anatomy knowledge acquired during the undergraduate course. The data was analyzed on Minitab 16 statistics package.

No statistical difference was observed between the grades of the two groups on the undergraduate topographic anatomy course. League members had a better performance on the research project examination. This difference was markedly observed between the more than 1‐year long members and the same grade nonmember students.

A still better performance was observed for students who had one full year of activities on the League as compared to students from the same grade that did not, suggesting that it may be an important element for knowledge fixation.
Explorative learning approach of problem‐based research project, a pilot study in anatomy and physiology course (721.4)
Authors: Hilvano Nestor
Published: June 2021

Teaching basic sciences has been approached through various teaching techniques aimed to facilitate learning and to improve student’s engagement. Oftentimes the students were presented with a specific case study, which lead the students to discuss through analysis of data supported by independent research and learning in order to present a diagnosis. An explorative learning approach was introduced in the Anatomy and Physiology course in which students were asked to use clinical skills and reasoning to create a patient case. Either individually or a team of 2‐4 students, they were given 5 weeks to submit a written problem‐based research project (PBRP) of student created patient case and a storyboard template of visual‐digital tool presentation. Rubrics for evaluation were developed for assessment and a survey tool was administered to rate the level of confidence on the various sections of a patient case write up and oral presentation. This pilot study shows that explorative learning in which students are asked to work backwards to generate a patient case write up and presentation of visual‐digital tool can be effective way of introducing students the skills needed to: a) promote active engagement in learning, b) provide learners considerable latitude in deciding on how to solve the problem, and c) achieve a greater understanding for the relevance of anatomy and physiology in clinical setting and practice.
Annotations enrich the virtual microscope slide experience (725.16)
Authors: Cotter John
Published: June 2021

PURPOSE: The uninitiated are often confused by the complexity of a histological specimen. And in the search for structures they are asked to identify, they may be so deterred by the task of locating and identifying structures they abandon the process for what they perceive to be easier ways of learning the organization of the organs and tissues. A solution may possibly lie in combining the virtues of a labeled atlas with the ease of viewing, panning and magnifying virtual slides (VS): at first, the users are guided to examples of the structures or features of organs they are to identify ensuring that nothing of importance is overlooked; and, with this introduction to the slide, the users can then explore the complexity of the VS by identifying structures on their own, with peers and/or with the assistance of an instructor while all the time being able to hide and show labels on the slide for reference purposes. METHODS: The VS are linked to LAYERS and LAYER REGIONS of the ANNOTATIONS box of the Aperio ImageScope viewer. And, selecting a structure listed in the LAYERS REGION moves the slide to an example of the structure. The hierarchy categorizes topics associated with the VS and indexes the objects to be identified. Characterization of the LAYERS allows users to explore a range of topics and hide and show the annotations for one or more topics. RESULTS: Positioning graphics and text with precision is problematic and there are spatial restraints viewing the ANNOTATIONS box and VS together. But, single VS can be used for many topics. And, students report being connected to the information, working efficiently and being deeply engaged (in contrast to examining microscope slides). The certainty of being directed to the structures is especially appealing. CONCLUSIONS: Virtual slides being facsimiles pose obstacles posed by glass slides whenever specimens are viewed by the untrained eye. The capacity to label VS and link to other resources offers a solution to the obstacles inherent in learning from unlabeled specimens.
The utilization of positive psychology and multiple modalities in the treatment of mentally ill persons (728.17)
Authors: Cartier William , Cartier Earnestine , Einstein George , Tulp Orien
Published: June 2021

To determine the contributions of positive psychology along with the integration of multiple behavioural modalities such as Cognitive Behavioral Therapy [CBT], Rational Emotive Behavioral Therapy [R.E.B.T], Individual and group psychotherapy were integrated with a positive orientated interdisciplinary team. Scientist and researchers have barely determined the full capabilities of the human brain and its unknown/hidden powers in terms of commanding/healing of the human body. It became apparent to our research team early on with correctional offenders, that “traditional” patient therapy simply wasn’t working and hence, the offender patient was not presenting any positive results or benefits for them. This study will present several case studies along with other data to illustrate the positive results obtained from correctional offender patient treated with positive results, utilizing the therapies/modalities mentioned above. This has been 5 years of intensive research (phase I) to determine a possible solution to our question. The next phase (phase II) will require 2 years of further intensive research. This will include research utilizing bio‐feedback, neuro‐biofeedback, Imaging (MRI), CAT and PET scan data. Additional research in this area is demanded along with dedicated efforts on the part of multidisciplinary research teams for years to come. Our changing world environment demands this of all of us define ourselves as research scientist when investigating the nature and outcomes of such client populations.

Grant Funding Source: Institutional Resources
Gene expression in breastmilk cells changes with the stage of lactation (732.2)
Authors: Hassiotou Foteini , Twigger Alecia‐Jane , Blancafort Pilar , Geddes Donna
Published: June 2021

Breastmilk contains a cellular hierarchy including pluripotent stem cells, progenitor cells, and differentiated epithelial cells and leukocytes. The latter are strongly associated with the health status of the mother/infant dyad. Little is known about the variation in the content of non‐leukocytic breastmilk cells. In this study, we used RT‐PCR to examine expression of genes representative of different cell types in breastmilk from over 50 healthy dyads from Australia and the USA, and the effects of lactation stage. Pluripotency genes were expressed at varying levels, with higher expression earlier in lactation. Expression of ESRRB, a pluripotency marker and reprogramming factor, was consistently higher than the other pluripotency genes, and than embryonic stem cells. The mammary stem cell marker CD49f was also highly expressed across all lactation stages. Expression of all stem cell genes tested was minimal in resting breast cells. These findings reinforce reports of stem cell gene expression in breastmilk cells across lactation, and suggest that some, but not all, of these genes are differentially regulated during lactation. These data now begin to elucidate developmental changes occurring in the breast during different lactation stages. They also ascertain an optimal window for accessing different breastmilk stem cell subpopulations for use in lactation research and regenerative medicine.
Overexpression of human polymerase ν in E. coli (736.2)
Authors: Shissler Susannah , Jiang Xiaohua
Published: June 2021

The human genome has a vast repertoire of proteins involved in replication and repair of DNA. Among those proteins, there are 15 total polymerases, and polymerase ν (pol ν) is one of the least studied. It is a member of A‐family polymerases, which are highly conserved. Studies of human polymerase ν have described a significantly higher error rate due to lack of proof‐reading capabilities. Because of this, pol ν is highly regulated within the cell to prevent extraneous mutation. Although pol ν has been shown to participate in translesion synthesis in vitro, its precise function has yet to be determined. Our Research with pol ν will include subcloning the catalytic core into the vector pBG106. The plasmid will be transformed into E. coli cells and the protein will be over‐expressed. Histidine‐tagged pol ν should simplify the separation and purification of the protein by use of a Nickel affinity column.
Kinases Kin1 and Kin2 pleiotropically modulate splicing and translation of HAC1 mRNA (746.2)
Authors: Anshu Ashish , Mannan M. Amin‐ul , Dey Madhusudan
Published: June 2021

Most nascent proteins fold and mature in the endoplasmic reticulum (ER). From the ER, matured proteins are packed inside a membrane‐bound vesicle that buds from the ER, travels and fuses with their target destination, thereby delivering the cargo protein. Stresses that disrupt ER function cause an over‐accumulation of un‐folded proteins in the ER, a state referred to as ER stress. To adapt to ER stress, cells activate a network of signaling pathways collectively called the unfolded protein response (UPR). The UPR in yeast Saccharomyces cerevisiae is initiated by a dual kinase/RNase Ire1 that removes an inhibitory intron from HAC1 pre‐mRNA, resulting in expression of Hac1 protein, a transcriptional activator of several UPR genes involved in adapting to ER stress. We have discovered that protein kinases Kin1 and Kin2 are dosage suppressors of a HAC1 allele that is devoid of a consensus element in the 3’ un‐translated region of pre‐mRNA. We have further discovered that both splicing and translation of HAC1 mRNA are reduced in a strain lacking kin1 and kin2 kinases, suggesting that the Ire1‐Hac1 signaling pathway is pleotropically regulated by Kin kinases. These Kin kinases have been reported to regulate polarized exocytosis and vesicular transport of proteins in yeast cells. Thus, our results revealed a novel role of Kin kinases in the yeast UPR.
Regulation of mitochondrial fission by MiD51 (757.1)
Authors: Losn Oliver , Rome Michael , Kaiser Jens , Shan Shu‐ou , Chan David
Published: June 2021

Regulation of mitochondrial fission by MiD51
Oliver C. Losόn1, Michael E. Rome1, Jens T. Kaiser2, Shu‐ou Shan2 and David C. Chan1,3
1Division of Biology and Biological Engineering, 2Division of Chemistry and Chemical Engineering and 3Howard Hughes Medical Institute, California Institute of Technology, Pasadena CA, 91125, USA Mitochondrial fission requires the recruitment of dynamin‐related protein 1 (Drp1) to mitochondria and activation of its GTP‐dependent scission function. In mammals, the recruitment of Drp1 is primarily regulated by the outer membrane receptors Mff, MiD49 and MiD51, while Fis1 plays a minor role. We recently demonstrated that these receptors can function independently of one another to recruit Drp1 and mediate mitochondrial division. Intriguingly, exogenous expression of MiD49 and MiD51 causes constitutive recruitment of Drp1 but inhibits its scission activity. With mitochondrial stress, however, these receptors can rapidly activate pre‐recruited Drp1 and cause rapid mitochondrial fragmentation independently of Fis1 and Mff. This behavior suggests an ability to sense mitochondrial function, but the molecular mechanism is unknown. To better understand the role of the MiDs in regulating mitochondrial fission, we produced recombinant protein pertaining to the cytosolic domain of each MiD, and performed crystallographic screens. We obtained crystals for MiD51 that diffracted to high resolution, and we were able to determine its structure. MiD51 possesses a nucleotidyl transferase fold and uses a variant set of residues to tightly bind a ligand. We have performed a structure‐function analysis of MiD51 by mutating structurally implicated residues and studying their importance in vivo and in vitro to activate Drp1. Our analysis reveals how MiD51 activates mitochondrial fission during mitochondrial stress.
Structure and evolution guided investigations of terpene synthase mechanism (769.11)
Authors: Peters Reuben
Published: June 2021

Structure and evolution guided investigations of terpene synthase mechanism Reuben J. Peters Department of Biochem., Biophys. & Mol. Biol., Iowa State University Terpene synthases catalyze wonderfully complex reactions, involving cyclization and/or rearrangements of isoprenyl diphosphate precursors. These reactions are driven by carbocationic cascades, which can be initiated in two distinct ways. Class I terpene synthases catalyze heterolytic cleavage of an allylic diphosphate ester bond to initiate their reactions, while class II terpene synthases utilize protonation for the same purpose. Building on our recent determination of high‐resolution crystal structures for both classes of terpene synthases, along with the conservation pattern underlying the evolution of novel function, we are probing the underlying enzymatic structure‐function relationships. Our results have revealed the ability of single residues changes to dramatically remodel reaction outcome, highlighting the role of electrostatic effects and providing significant insights into the catalyzed complex reactions.

Grant Funding Source: GM076324
Structural and biochemical analysis of a type II free methionine‐R‐sulfoxide reductase from Thermoplasma acidophilum (774.7)
Authors: Kim Hwa‐Young , Kim Hyun Sook , Kwak Geun‐Hee , Lee Kitaik , Jo Chang‐Hwa , Hwang Kwang Yeon
Published: June 2021

Free methionine‐R‐sulfoxide reductase (fRMsr) enzymes only reduce the free form of methionine‐R‐sulfoxide (Met‐R‐O). These enzymes can be grouped into two types with respect to the number of conserved Cys residues in the active sites. Type I fRMsr enzymes, which have three conserved Cys residues, have been characterized biochemically and structurally. However, the structural and biochemical properties of type II enzymes, which have two conserved Cys residues, are unknown. Here, we present the crystal structures of type II fRMsr from Thermoplasma acidophilum in native form and in a complex with Met‐R‐O. T. acidophilum fRMsr functions as an antioxidant protein. Based on biochemical and growth complementation assays, Cys84 is demonstrated to be the catalytic Cys. Structural analyses also support the catalytic function of Cys84 and provide insights into substrate recognition and orientation, conformational changes in the active site during substrate binding, and the role of active site residues in substrate binding and catalysis. A model for the catalytic mechanism of type II T. acidophilum fRMsr is proposed, in which a resolving Cys is not involved and the catalytic Cys84 sulfenic acid is directly reduced by the reductant thioredoxin.
Designing assessment for learning: leave with one in hand (80.1)
Authors: O'Sullivan Patricia
Published: June 2021

We would like to move from designing assessments of learning to assessments for learning. In this session, we will first describe the differences between these two perspectives. Then, we will examine examples of current assessments to see how they might be transformed into an assessment for learning. Guidelines will be provided so that participants will be able to transform their own assessments.
Genistein‐induced histone modifications on ATF3 may contribute to cell cycle arrest and apoptosis in human colon cancer cell line SW620 (800.8)
Authors: Chen Danni , Li Qian , Chen Hong
Published: June 2021

Genistein, a soy‐derived isoflavone, has been shown to suppress colon cancer progression by regulating different cellular properties. The aim of the study is to investigate the role of Activating Transcription Factor 3 (ATF3), an immediate‐early response gene, in genistein‐modulated cell cycle arrest and apoptosis in a human colon cancer cell line SW620. Results showed that genistein induced G2 phase arrest and increased late apoptosis in SW620. Genistein also increased the expression of ATF3 mRNA and protein, as well as its transcriptional activity in SW620. mRNA expression of downstream genes of ATF3, such as cell cycle inhibitors p53, p21, CyclinD1, and apoptotic enzymes Casp8, and Casp9, were all induced by genistein. Chromatin immunoprecipitation (ChIP) showed decreased phosphorylated histone H3 serine 10 (H3S10p) at both the promoter and coding region of ATF3. In addition, genistein increased trimethylated histone H3 lysine 36 (H3K36me3) level at a coding region and dimethylated histone H3 lysine 4 (H3K4me2) level at the promoter region. Methylation sensitive PCR showed that genistein did not change the level of DNA methylation at various regions of ATF3 gene. In conclusion, genistein‐mediated histone modifications of ATF3 may lead to G2 phase arrest and apoptosis in SW620.
G‐protein beta‐gamma regulation of Golgi to cell surface transport (802.11)
Authors: Klayman Lauren , Wedegaertner Philip
Published: June 2021

Non‐canonical G‐protein signaling has been shown to regulate intracellular transport and secretion. G‐protein beta‐gamma localizes to the trans‐Golgi network and activates Protein Kinase D (PKD), resulting in fission of vesicles destined for the plasma membrane. While basolaterally‐targeted (BL) or secretory vesicle release is known to be downstream of PKD activation, apically‐targeted (AP) vesicle release from the Golgi occurs via a PKD‐independent pathway. A similar cargo selectivity for beta‐gamma has not been tested. Here, we generate GRK2ct variants that inhibit beta‐gamma in a spatial or temporal manner. We test regulation of AP and BL cargo trafficking via a temperature shift immunofluorescence transport assay, as well as trafficking of other AP and BL candidate cargoes. We find that lipid‐association of the GRK2ct is critical for its inhibitory function on beta‐gamma. We also generate an inducible system of recruitment of GRK2ct variants to distinct subcellular locations, towards the goal of inhibiting solely Golgi‐localized beta‐gamma signaling. Our system can also be used to inhibit beta‐gamma signaling at other endomembrane locations. The results indicate that beta‐gamma is an upstream regulator of vesicle transport from the Golgi to the cell surface.

Grant Funding Source: NIH GM056444
Characterization of tumor‐derived B56γ mutations and their effect on the tumor suppressor function of B56γ‐PP2A (802.21)
Authors: Nobumori Yumiko , Liu Xuan
Published: June 2021

Protein phosphatase 2A (PP2A) is a family of serine/threonine phosphatases with diverse cellular functions. Previous study reported that B56γ subunit directs PP2A toward tumor suppressive function by dephosphorylating and activating tumor suppressor p53. B56γ‐PP2A also inhibits cell transformation by an unknown p53‐independent mechanism. In this study, we investigate the inactivation mechanism of B56γ tumor suppressive function through characterization of B56γ mutations identified in human cancer. The first class mutations disrupt interaction between B56γ and AC core, and thus B56γ fails to regulate its substrates and completely lose tumor suppressive function. Further, we show that C39R mutation within HEAT repeat1 disrupts B56γ binding with AC core, providing structural insight that intact HEAT repeat1 is required for B56γ‐PP2A holoenzyme assembly. The second class mutations prevent specific substrates from binding B56γ and cause partial reduction of tumor suppressive function of B56γ. We find that A383G and F395C mutations disrupt B56γ from interacting p53. Mutants are unable to dephosphorylate p53 and thus lack p53‐dependent tumor suppressive function, implying that bridging interaction between B56γ and p53 is required for p53 activation by B56γ‐PP2A. In addition, we identified S220N mutation that specifically disrupts p53‐independent function of B56γ, suggesting that it may disrupt the binding of unknown substrates. Although it remains to be investigated how frequently these mutations occur, our results underline the importance of B56γ tumor suppressive function in human cancer.

Grant Funding Source: Supported by NIH
Post‐phosphorylation control of dopamine transporter by peptidyl prolyl cis‐trans isomerase PIN1 (803.6)
Authors: Challasivakanaka Sathyavathi , Smith Margaret , Foster James , Vaughan Roxanne
Published: June 2021

The rat dopamine transporter (DAT) is phosphorylated at the N‐terminus on T53, a site that is followed by proline, making it specific for proline‐directed kinases such as ERK. We found that amphetamine (AMPH) stimulates DAT T53 phosphorylation and mutating this site affects DA transport and AMPH‐stimulated efflux, suggesting a role for T53 phosphorylation in these processes. Phosphorylation of proline‐directed residues strongly impacts protein conformation by reducing the rate of cis‐trans isomerization of the phospho (p)‐S/T‐P peptide bond. This in turn slows the rate of p‐S/T‐P dephosphorylation by preventing actions of phosphatases. The peptidyl prolyl cis‐trans isomerase, PIN1, specifically isomerizes p‐S/T‐P motifs and catalyzes the return of the bond to the trans configuration allowing dephosphorylation to occur. To test for such activities on DAT we have used the PIN1‐specific inhibitor juglone. In both rat striatal synaptosomes and heterologous expression systems juglone treatment rapidly increased the p‐T53 level suggesting that PIN1 acts on DAT and regulates dephosphorylation of T53. In addition, juglone strongly stimulated [3H]DA efflux from rat striatal synaptosomes, supporting a mechanistic link between efflux and p‐T53. These results identify a novel mechanism for regulation of DA efflux and suggest PIN1 as an enzyme that functions in post‐phosphorylation control of DAT function.

Grant Funding Source: Supported by NIDA R01 DA 13147
Will altering visual cues of food consumption affect food intake in preschool‐age children? (808.8)
Authors: Kranz Sibylle , Brauchla Mary , McCabe Sean , McCabe George
Published: June 2021

Portion size is known to influence food intake in children, but little is known about how visual cues will affect food consumption in this age group. This crossover study tested whether children’s food intake would increase if children had no visual cues (NVC) of food consumption. Two‐to‐five year old children (n=38) were asked to come to Purdue University on Saturday mornings for 6 visits. After a snack preload, children were served one of 3 lunch foods: pasta with sauce, yogurt, or high‐fiber yogurt. During one visit, children had standard visual cues of food consumption and at the other visit there were no visual cues (refilling bowl). A repeated measures ANOVA model was used to assess the effects of visual cues, gender, food and their interactions on food consumption. Contrasts were used to assess the effect of visual cues on food consumption. We found no significant effects of visual cues on food intake in any of the study foods, although there was a nonsignificant trend towards greater intake in the NVC condition with two of the three foods tested. Food intake in preschoolers may not be affected by visual cues of amount consumed, but additional research is needed to further test this hypothesis.
Over‐the‐counter prenatal multivitamin/mineral products: chemical analysis for the Dietary Supplement Ingredient Database (809.3)
Authors: Gusev P , Andrews K , Palachuvattil J , Dang P , Roseland J , Holden J , Savarala S , Pehrsson P , Dwyer J , Betz J , Saldanha L , Bailey R , Costello R , Gahche J , Hardy C , Emenaker N , Douglass L
Published: June 2021

Many women use OTC prenatal MVM products to optimize nutrient intake before pregnancy, during pregnancy and lactation. Most prenatal MVM are purchased as prescription products, but the OTC market has increased, especially in the natural health and direct sales channels. Using a statistical sampling plan, 71 DS products were purchased, with 16 from mass market, 45 from natural health and 33 from direct channels. Multiple lots of each product were sent to qualified labs with quality control materials for the analysis of vitamins and minerals. Analytical results were compared to label and evaluated by regression across a range of labeled levels. The predicted mean % differences from label at the most common level ranged from 0 to <10% above label for 12 nutrients (iron, magnesium, zinc, copper, selenium, manganese, riboflavin, folic acid, vitamins A, B‐12, C, E); 10 to <20% for vitamin D and calcium; 20 to <30% for iodine and potassium, and >40% for chromium. The predicted mean % differences for thiamin, niacin and vitamin B‐6 were in the ‐10 to 0% range. For most ingredients, predicted mean analytical values are similar to and above the label, but some are significantly different. These data will be released in DSID‐3 (http://dsid.usda.nih.gov) and can provide researchers with more accurate estimates of nutrient intake from OTC prenatal MVM.

Grant Funding Source: Supported by USDA‐ARS and NIH‐ODS
Achievements and discoveries as to functional foods in Japan (813.14)
Authors: Kumagai Hitomi
Published: June 2021

The term and concept of “functional foods” were created in the course of “Grant‐in‐Aid” research sponsored by the Japanese Ministry of Education, Science and Culture in 1980s. Functional foods are defined as those that modulate body function and prevent certain diseases. Since then, numerous studies have been conducted as to the physiological functions of food factors. The Japanese Ministry of Health and Welfare established a policy to approve some of the selected functional foods as “foods for specified health uses (FoSHU)” in 1991 to share the benefit of results obtained by the research works and provide people with healthy foods manufactured on the basis of scientific achievements. At present, the Customers Affairs Agency is responsible for the approval, and over 1,000 items have been approved as FoSHU. FoSHU products are categorized into 10 according to their functions such as modulation of gastrointestinal condition, serum cholesterol level, serum triacylglycerol level, blood pressure and blood sugar level, acceleration of mineral absorption and promotion of bone health. Interestingly, functional factors in products of the same function often differ in their mechanism of action, while the same functional factor is contained in products of different functions. Therefore, this presentation will summarize some of the functional factors according to their properties and characteristics to get a hint for the strategy to design novel functional foods.
Intestine‐specific inactivation of acyl CoA: monoacylglycerol acyltransferase 2 protects mice from diet‐induced obesity (822.12)
Authors: Yen Chi‐Liang Eric , Nelson David , Gao Yu , Yen Mei‐I
Published: June 2021

Acyl: CoA: monoacylglycerol acyltransferase (MGAT)2 catalyzes the re‐esterification of triacylglycerol in the enterocytes, a step required for fat absorption. Mice without functional MGAT2 enzymes (Mogat2‐/‐) are protected from diet‐induced obesity but, surprisingly, absorb normal amount of dietary fat and exhibit increased energy expenditure. MGAT2 is expressed in tissues besides intestine, including the kidney and adipose tissues in both mice and humans. To test the hypothesis that intestinal MGAT2 regulates systemic energy balance, in this study, we generated and characterized mice deficient in MGAT2 exclusively in the small intestine (Mogat2IKO). We found that, like Mogat2‐/‐ mice, Mogat2IKO mice showed delayed in fat absorption, a transient decrease in food intake when first exposed to a high‐fat diet, and a propensity to use fatty acids as fuel. Though to a lesser extent than Mogat2‐/‐ mice, Mogat2IKO mice also exhibited a persistent increase in energy expenditure and are protected against diet‐induced weight gain, excessive expansion of body fat, and associated comorbidities, including hepatic steatosis, hypercholesterolemia, and glucose intolerance. These findings illustrate that intestinal lipid metabolism plays a crucial role in the regulation of systemic energy balance. In addition, they also suggest that MGAT activity in tissues beside intestine may also contribute to energy metabolism.

Grant Funding Source: Supported by NIH DK088210
Impact of processing on the bioavailability and vascular effects of blueberry (poly)phenols (831.12)
Authors: Rodriguez‐Mateos Ana , George Trevor , Heiss Christian , Spencer Jeremy
Published: June 2021

Blueberries are a rich source of flavonoids and phenolic compounds. Currently, little information is available regarding the effects of processing on the bioactivity and bioavailability of berry (poly)phenols. Two randomized, controlled crossover trials were carried out to assess the effects of a blueberry drink and a blueberry baked product on endothelial function measured as flow‐mediated dilation (FMD), and in (poly)phenol absorption and metabolism in healthy individuals. Significant increases in %FMD at 1, 2 and 6 hours after consumption of both blueberry products were observed when compared with controls. The changes were correlated with the appearance of plasma blueberry (poly)phenol metabolites. Although some changes were observed on the bioavailability of blueberry (poly)phenols after processing, blueberry delivered as a drink or as a baked product improved endothelial function in healthy volunteers.

Grant Funding Source: Alpro Foundation
The roles of thromboxane A2 in high salt‐induced defect during pregnancy (832.4)
Authors: Lin Shu‐Wha , Pai Chen‐Hsueh , Yen Ching‐Tzu , Yu I‐Shing , Lin Shu‐Rung
Published: June 2021

Preeclampsia is a pregnant‐related hypertensive disorder. Affected women might be at risk of developing eclampsia that would cause fetal and maternal death. Several studies have indicated that women suffering from preeclampsia with high levels of thromboxane A2 (TXA2) metabolites in plasma and urine might induce thrombosis and hypertension, and developmental retardation of the placenta. We had clinical data indicating that thromboxane A2 synthase (TXAS) were highly expressed in preeclamptic placenta. To investigate the roles of TXA2 in pregnancy‐related hypertension and preeclampsia, pregnant WT mice and TXA2deficient (TXAS KO) mice were fed with high salt in drinking water. We found that high salt‐treatment induced more severe syndromes in WT than in TXA2‐deficient pregnant mice. Hypertension characterized by significantly increased blood pressure was observed in high salt‐treated WT but not TXA2‐deficient pregnant mice. Compared to normal nonpregnant female mice fed with high salt, pregnant mice with the same treatment had severely decreased maternal body weight, fetal size and fetal weight. High salt feeding also induced cellular apoptosis and suppressed cellular proliferation in the placenta of pregnant but not nonpregnant mice. Moreover, increased abnormal cell degeneration was also observed in the fetuses of WT but not TXA2‐deficient mice. These results demonstrate that deficiency of TXA2 in pregnant mice rendered mice more resistant to high salt‐induced hypertension and growth retardation of placenta and fetus, suggesting a therapeutic potential of inhibitors of the thromboxane pathway for preeclampsia.
Characterization of vascular smooth muscle cell function in Gravin (AKAP12) truncated mice model in response to angiotensin‐II (866.7)
Authors: Fan Qiying , Wijaya Cori , Diaz Diaz Andrea , McConnell Bradley
Published: June 2021

Vascular smooth muscle cells (VSMCs), plays a crucial role in the progression of atherosclerosis in response to angiotensin II (Ang II). Gravin, one of the A‐kinase anchoring proteins, targets protein kinase A (PKA), protein kinase C (PKC), and other molecular to different cellular compartments and coordinates signal transduction. The purpose of this study is to investigate the role of gravin in mediating Ang II signaling pathway in VSMCs during atherosclerosis using gravin truncated (gravin‐t/t) mice. Our data showed that Ang II ‐induced migration and proliferation of VSMCs was decreased in gravin‐t/t mice when compared to wild type (WT) mice. Furthermore, gravin‐t/t VSMCs showed no significant difference in PKA activities with or without Ang II treatment, but exhibited a decreased PKC activities and lower intracellular Ca2+ transit after Ang II treatment. These changes were accompanied by significant differences in PKC and PKC‐dependent substrate phosphorylation involving ERK1/2 signaling pathway. In particular, gravin‐t/t mice showed a decreased expression of matrix metalloproteinase 9 (MMP9, a marker of atherosclerosis) and proliferative cell nuclear antigen (PCNA) versus WT mice in the aorta. In conclusion, these findings demonstrate that gravin regulates Ang II ‐induced VSMC proliferation/migration, indicating gravin’s role in the progression of atherosclerosis and/or vascular remodeling.
Glucosamine attenuates cigarette smoke‐induced lung inflammation via its antioxidant function and by inhibiting ROS‐sensitive inflammatory signaling (869.12)
Authors: Lin An‐Hsuan , Wu Yuh‐Lin , Liu Meng‐Han , Lee Tzong‐Shyuan , Kou Yu Ru
Published: June 2021

Cigarette smoking causes persistent lung inflammation that is mainly regulated by redox‐sensitive pathways. We have reported that cigarette smoke (CS) activates a NADPH oxidase‐dependent reactive oxygen species (ROS)‐sensitive AMP‐activated protein kinase (AMPK) signaling pathway leading to induction of lung inflammation. Glucosamine, a dietary supplement used to treat osteoarthritis, has antioxidant and anti‐inflammatory properties. However, whether glucosamine has similar beneficial effects against CS‐induced lung inflammation remains unclear. Using a murine model we show that chronic CS exposure for 4 weeks increased lung levels of 4‐hydroxynonenal (an oxidative stress biomarker), phospho‐AMPK, and macrophage inflammatory protein 2, and induced lung inflammation; all of these CS‐induced events were suppressed by chronic treatment with glucosamine. Using human bronchial epithelial cells (HBECs), we demonstrate that cigarette smoke extract (CSE) sequentially activated NADPH oxidase; increased intracellular levels of ROS; activated AMPK, mitogen‐activated protein kinases (MAPKs), nuclear factor‐κB (NF‐κB), and signal transducer and activator of transcription proteins 3 (STAT3); and induced interleukin‐8 (IL‐8). Additionally, using a ROS scavenger, a siRNA that targets AMPK, and various pharmacological inhibitors, we identified the signaling cascade that leads to induction of IL‐8 by CSE. All these CSE‐induced events were inhibited by glucosamine pretreatment. Our findings suggest a novel role for glucosamine in alleviating the oxidative stress and lung inflammation induced by chronic CS exposure in vivo and in suppressing the CSE‐induced IL‐8 in vitro via its antioxidant function and by inhibiting both ROS‐sensitive NADPH oxidase/AMPK/MAPK signaling pathway and the downstream transcriptional factors NF‐κB and STAT3.
Effect of Q‐VD‐OPh on neurodegeneration and neuroinflammation of sarin‐exposed mice (877.6)
Authors: Shah Ekta , Grunwald William , Garrett Teresa , Lucot James , Brown Thomas , Cool David
Published: June 2021

Organophosphates (OP) cause CNS hyperstimulation resulting in seizures, convulsions and brain damage. Sarin is a toxic nerve agent used in Syria, the Gulf War and in terrorist attacks in Japan. Exposure to sarin is associated with presence of dead neurons, activation of astrocytes, microglia and NI. Since, cell death and NI are found in sarin exposed animals, we evaluated the efficacy of Q‐VD‐OPh, a pan spectrum caspase inhibitor that prevents cell death and has anti‐inflammatory properties. Currently, treatment for sarin exposures is effective only if administered within 30‐40 minutes of exposure. In this study, adult female C57BL/6 mice were subjected to an injection of 1.5 mg/kg CBDP before a 0.04mg/kg injection of sarin followed by 20 mg/kg injection of Q‐VD‐OPh 30 minutes later. Mice were sacrificed at two and fourteen day time points followed by BioPlex (Bio Rad) cytokine analysis of amygdala, hippocampus and piriform cortex. At two days, there was a significant increase (p<0.05) in IL‐2, IL‐10, IL‐1b, MIP‐1b and MCP‐1in sarin exposed mice, and a significant decrease in the same when injected with Q‐VD‐ OPh. Q‐VE‐OPh did not appear to have an effect. The data strongly suggests that Q‐VD‐OPh provides anti‐inflammatory properties in mice exposed to sarin. This project was supported by a grant from the NIH R21NS072103‐03. Dr. Cool and Dr. Brown have an equity interest in Apoptrol, LLC (Vice President and President, respectively) which supplied the Q‐VD‐OPh and Q‐VE‐OPh for the study.

Grant Funding Source: NIH R21NS072103‐03