bluuebunny/crossref_metadata_2025 huggingface dataset:
Introduction
Authors: Boyle A.J.
Published: June 2021

oratio certam regulam non habet; consuetudo illam ciuitatis, quae numquam in eodem diu stetit, uersat.

Style has no fixed rules; the usage of society changes it, which never stays still for long.

Seneca Epistle 114.13

This is the first of two volumes of critical essays on Latin literature of the imperial period from Ovid to late antiquity. The focus is upon the main postclassical period (A.D. 1-150), especially the authors of the Neronian and Flavian principates (A.D. 54-96), several of whom, though recently the subject of substantial investigation and reassessment, remain largely unread, at best improperly understood. The change which took place in Roman literature between the late republic/early Augustan period and the post-Augustan empire, between the ‘classicism’ of Cicero, Virgil, Horace, Livy and the ‘postclassicism’ of Seneca, Lucan, Persius, Tacitus is conventionally misdescribed (albeit sometimes with qualifications) as the movement from Golden to Silver Latin. The description misleads on many counts, not least because it misconstrues a change in literary and poetic sensibility, in the mental sets of reader and audience, and in the political environment of writing itself, as a change in literary value. What in fact happened awaits adequate description, but it seems clear that the change began with Ovid (43 B.C. to A.D. 17), whose rejection of Augustan classicism (especially its concept of decorum or ‘appropriateness’), cultivation of generic disorder and experimentation (witness, e.g., Ars Amatoria and Metamorphoses), love of paradox, absurdity, incongruity, hyperbole, wit, and focus on extreme emotional states, influenced everything that followed. Ovid also witnessed and suffered from the increasing political repression of the principate; he was banished for — among other things — his words, carmen. And political repression seems to have been a signal factor, if difficult to evaluate, in the formation of the postclassical style.
New Ideas for the Development of Tea Culture Tourism Economy in the Internet Age
Authors: Deng Fei
Published: June 2021

Tea originated in China and flourished in China. Tea has a long history for China. Tea has given birth to rich history and culture in the long river of history. Tea culture has become an important part of our excellent traditional culture, which has given birth to countless traditional cultural industries. Although with the development of the times, Chinese traditional cultural industry is facing many challenges. Fortunately, the arrival of the information age has brought new opportunities for the traditional cultural industry. This paper analyzes the new ideas brought by the era of “Internet” for the development of tea culture tourism economy, and discusses how to better use information technology to help the development of traditional cultural industry.
GDP forecast of accommodation and catering industry in X city based on multi variable grey model —— Energy saving development of accommodation and catering industry
Authors: Gao Qian , Liu Yunyun , Yang Junyi , Hong Yu , Wen Lei
Published: June 2021

As the GDP of lodging and catering industry is generally quarterly or annual, the data volume is very small, so it is difficult to accurately predict the GDP of lodging and catering industry in this city for a city. However, a city can accurately predict the GDP development of the lodging and catering industry through electricity consumption. And the local government can formulate relevant policies to promote the stable development of lodging and catering industry and economy, and find the relationship between electricity consumption and GDP development of this industry, so as to achieve the purpose of low energy consumption and fast economic growth.t. This paper first conducted desensitization and normalization processing on the data, and then used Matlab to forecast the GDP of the lodging and catering industry in X City respectively by using GM (1, N) grey model, multiple linear regression model and BP neural network model. The experimental results show that the GM (1, N) grey model is more accurate and reliable than the other two methods in the case of less data. This paper solves the problem that the GDP of accommodation and catering industry in a single city is inaccurate.
Klinische und praktische Erfahrungen mit Lymphstrategien bei Zivilisationserkrankungen
Authors: Matejka Rainer
Published: June 2021

Das gesamte System der Lymphe wird immer noch unterschätzt. Dabei spielt es in allen Bereichen des menschlichen Stoffwechsels eine wichtige Rolle, sowohl bei der Zellausscheidung als auch bei der Immunaktivität. Vor allem die modernen Zivilisationskrankheiten sollten im Hinblick auf die Lymphe bedacht und mitbehandelt werden. Der Beitrag unterstreicht die Bedeutung der Lymphe bei diversen Erkrankungen und Beschwerdebildern und stellt lymphrelevante Behandlungsansätze für Adipositas und chronisch-entzündliche Darmerkrankungen vor.
GLYCINE/SARCOSINE RATIO AS NOVEL BIOMARKER FOR ALCOHOL‐INDUCED LIVER FIBROSIS UNDER SUMOYLATION CONTROL
Authors: Tomasi Maria Lauda , Cossu Carla , Ramani Komal , Floris Andrea
Published: June 2021

Propose

Alcohol‐induced liver fibrosis/disease (ALD) is characterized by excessive deposition of extracellular matrix (ECM) components in response to chronic abuse that could lead to cirrhosis and hepatocellular carcinoma development. Sarcosine is a derivative of the amino acid glycine, formed by the enzymes glycine N‐methyl transferase (GNMT) or dimethylglycine dehydrogenase (DMGDH) and converted back into glycine via sarcosine dehydrogenase (SARDH). GNMT is silenced in human alcohol‐induced cirrhosis. In addition, GNMT knockout mice develop oxidative stress, liver injury, fibrosis, and HCC. SUMOylation is a post‐translational modification that requires an essential E2‐conjugating enzyme 9 (UBC9) to covalently bind of small ubiquitin modifier (SUMO) and plays an important role in a wide range of cellular processes. We previously demonstrated that UBC9 level is induced in intragastric ethanol‐infusion (EI) treated mouse liver. We performed SUMO‐proteomics of alcohol‐fed mouse liver and identified altered sumoylation of GNMT and SARDH. The goal of this work is to examine whether the dysregulated SUMOylation could regulate GNMT and SARDH enzymatic function in ethanol‐induced liver fibrosis and elucidate the molecular mechanism(s).

Methods

Studies were performed using mouse hepatocytes (HEP), kupffer cells (KCs) and stellate cells (HSCs) from in vivo acute and chronic ethanol‐fed mouse models and primary human HSCs. Oxidative stress and metabolic markers were measured using commercial kits.

Results

We found that ethanol treatment in vivo induced UBC9 expression in HSCs and HEP and inhibited its expression in KCs. This was associated with increased GNMT sumoylation and total levels, specifically in HSCs. In contrast, SARDH sumoylation fell in HSCs despite an increase in its total level. Ethanol feeding increased glycine and lowered sarcosine levels in mouse plasma, suggesting a potential regulation of GNMT/SARDH enzyme activity. Ethanol‐treated co‐culture model (HEP, HSCs and KCs) showed increase in ROS production in all liver cells and glycine/sarcosine ratio in HSCs but not in HEP and KCs compared to control. Ubc9 silencing in HSCs inhibited ethanol‐mediated ROS production and induction of HSC activation.

Conclusions

This data strongly suggests that oxidative stress‐induced sumoylation plays a key role in HSC activation and this may influence the transmethylation machinery. Alterations of glycine/sarcosine ratio as a consequence of enzyme sumoylation could be considered as novel biomarker for ALD.

Support or Funding Information

NIAAA 5 K01 AA022372‐06

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neural Crest Derived Cardiomyocytes Regulate Cardiac Trabeculation and Adult Heart Function.
Authors: Abdul‐Wajid Sarah , Demarest Bradley , Yost H Joseph
Published: June 2021

The cardiac neural crest arise from the multi‐potent, migratory neural crest cell population and contribute to cardiovascular development, including contributions to cardiac endothelium and myocardium, formation of the great vessels and their valves, as well as septation of the chambers. However, very little is known about the neural crest contribution to the myocardium. This study aims to selectively decipher the function of the neural crest derived cardiomyocytes (NC‐Cms) without the confounding roles of the contributions of neural crest to other tissues, including endocardium. Zebrafish provide a unique genetic model to study this population because of their developmental transparency and they have a relatively large neural crest contribution of NC‐Cms compared to mammals. By creating Zebrafish transgenic lines that both lineage label and allow for ablation of only NC‐Cms, we find that the embryonic NC‐Cms arrive early in zebrafish heart development, makeup ~10% of the embryonic cardiomyocyte population and are steady state by two days post fertilization (2dpf). A majority of this contribution is to the ventricle in a discrete pattern around the apex of the ventricle. To test the function of NC‐Cms during heart development, we genetically ablated them during their early stages (up to 2dpf) and then observed heart development and phenotype up to 14dpf and into adulthood. Surprisingly embryonic ablation of the NC‐Cms was largely dispensable for heart development with a non‐lethal but significant defect in patterning of ventricle trabeculation. Importantly, embryonic ablation of NC‐Cms yielded adults with hypertrabeculated hearts and impaired performance in cardiac stress tests. We investigated the relation of the NC‐Cms to known Notch and Neuregulin effectors of embryonic trabeculation and speculate on their mechanistic regulation of this pathway. These results present a new model of human left ventricular non‐compaction or hypertrabeculation cardiomyopathy, and adult heart failure. Importantly these results demonstrate how a subtle and relatively minor developmental cell population can have detrimental effects on adult cardiac function.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interactive Cases using the Medical Stories of a Living Anatomical Donor
Authors: Elzie Carrie , Wellman Laurie , Ortiz Fabian , Clark Russ M. , Goodmurphy Craig W.
Published: June 2021

Narrative medicine is an important and expanding new field that recognizes the value of storytelling in the acts of doctoring. Narrative competence involves recognizing, absorbing, metabolizing, interpreting and being moved by the stories of illness. Our goal was to incorporate this valuable aspect of medicine into the gross anatomy course with first year medical students. To do so, we have taken the illness stories of one living anatomical donor, Mr. Clark, and transformed them into interactive cases and discussions to teach not only the humanity of medicine, but also the anatomical sciences. Five digital interactive cases were generated combining Mr. Clark's personal narratives of his medical encounters, his own medical records and relevant anatomical content using Articulate Storyline. Students reviewed the cases prior to attending a discussion session with Mr. Clark facilitated by a physician where students were allowed to ask any questions which ranged from personal feelings to medical details to financial implications. Qualitative analysis has shown that the conversation has reflected more compassion and reflection than science and medicine. However, anatomical questions based on the cases have performed equally or better than the traditional anatomical course content. Anatomical reflections since the addition of Mr. Clark, who has participated in every anatomy lab and ultrasound session, have become more empathetic and less egoistic. Focus group feedback and course evaluations have been overwhelmingly positive and reviews have outranked similar cases using virtual patients. Thus we encourage all anatomy programs to reach out to prospective donors, as the addition of their stories, has truly enhanced our anatomy course and our students' learning.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Memantine (a dual α7‐nAChR/NMDAR antagonist) displays anti‐angiogenic activity in human squamous cell lung cancer
Authors: Nolan Nicholas A. , Robateau Zachary , Brown Kathleen C. , Chen Yi Charlie , Egleton Richard D. , Tirona Maria R. , Dasgupta Piyali
Published: June 2021

Cigarette smoking accounts for 85% of squamous cell lung cancer (SCC‐L) in patients. Since SCC‐L is a highly angiogenic tumor, anti‐angiogenic therapies like Avastin have been investigated for the treatment of SCC‐L. However, the drawback of these therapies has been the incidence of pulmonary hemorrhage and extensive hemoptysis in SCC‐L patients. Nicotine, the addictive component of cigarettes, promotes angiogenesis in lung cancer via the α7‐nicotinic acetylcholine receptor (α7‐nAChR) subunit in human lung endothelial cells. Therefore, we conjectured that α7‐nAChR‐antagonists should display potent anti‐angiogenic and antitumor activity in SCC‐Ls. We observed that the α7‐nAChR is robustly expressed on human microvascular endothelial cells of the lung (HMEC‐Ls) and SCC‐L associated endothelial cells (STACE). With this background in mind, we tested the anti‐angiogenic activity of memantine (dual α7‐nAChR/NMDAR antagonist) in SCC‐Ls. Receptor binding assays have shown that the affinity of Memantine for α7‐nAChR is greater than NMDAR. Here we show that memantine attenuates nicotine‐induced angiogenesis in human microvascular endothelial cells of the lung (HMEC‐Ls). Furthermore, the α7‐nAChR antagonist memantine displayed potent antiangiogenic activity in the chicken chorioallantoic membrane (CAM) model. Memantine is an FDA approved drug, used in clinical practice to treat mild‐moderate Alzheimer's disease. It is well tolerated by patients and has a favorable side effect profile. Our studies are unique and innovative because they study, for the first time, the application of memantine in a non‐neuronal disease, namely lung cancer.

Support or Funding Information

Funding for our study was supported by an NIH R15‐AREA Grant (2R15CA161491‐02), Edwards Cancer Center Pilot grant, Cabell Huntington Hospital, Marshall University. This work was supported in part by the West Virginia IDeA Network of Biomedical Research Excellence (WV‐INBRE) grant GM103434 (PI: Dr. G. Rankin).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Heterotopic Ossification in Fibrodysplasia Ossificans Progressiva – How Does One Tissue Become Another?
Authors: Shore Eileen M.
Published: June 2021

The development and maintenance of tissues and organ systems depend on precise coordination of cellular events and mechanisms. During embryonic development, the induction of bone formation is spatially and temporally regulated to specify the skeletal elements. After birth, new bone formation is normally limited to regeneration during fracture repair, a process that is also precisely regulated.

Fibrodysplasia ossificans progressiva (FOP) is a rare human genetic disease in which extensive and progressive heterotopic ossification – the formation of bone outside of the normal skeleton – occurs in soft connective tissues such as skeletal muscle. At birth, there is usually little indication of the disease, except for a characteristic malformation of the great toes. However during early childhood, episodes of bone formation that are frequently associated with swelling and inflammation begin within soft tissues. In most patients examined to date, FOP is caused by a single nucleotide change in the BMP type I receptor ACVR1 (Activin A receptor, type 1; alias, ALK2) that causes enhanced pathway activation. All patients with a classic clinical presentation of the disease possess a recurrent heterozygous c.617G>A (R206H) mutation.

Heterotopic ossification in FOP frequently forms in response to tissue injury, although also develops in the absence of overt tissue trauma. Within the tissue, heterotopic bone formation in FOP patients progresses through well‐described changes in affected tissue: initial catabolic events are followed by an anabolic stage and formation of endochondral bone. In vivo mouse models have been instrumental in providing a more detailed understanding of the tissue, cellular, and molecular impact of the ACVR1 R206H mutation and the structural changes in the soft connective tissues that are transitioning to cartilage and bone. In response to injury, initial steps of wound healing in mutant tissue appear to be normal, including an early immune response that leads to tissue degradation and removal of damaged tissue. However this tissue repair trajectory rapidly diverges and, instead of repairing and regenerating the injured muscle tissue, ectopic cartilage and bone are formed. Our investigations of these events are providing a more detailed understanding of the tissue, cellular, and molecular impact of the ACVR1 R206H mutation and of the mechanisms that regulate tissue maintenance, repair, and regeneration.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Postdoctoral Fellowship in Anatomy Education – Is it Needed?
Authors: Schaefer Audra
Published: June 2021

With an increasing demand for anatomy faculty members, a number of graduate programs have been implemented to train individuals to be able to teach the various anatomical disciplines. Such programs offer instruction in pedagogy, scholarly teaching and educational research. Although there are increasing numbers of masters and doctoral level programs that have emerged, anatomy education focused postdocs are nearly nonexistent. In this study, members of the American Association of Anatomists were surveyed to determine whether they perceived that a postdoctoral fellowship emphasizing educational research and pedagogy training in the anatomical sciences would be a worthy endeavor. Initially, a pilot survey was conducted, inviting 27 early to mid‐career anatomy faculty to participate. For the second administration of the survey, an open invitation was posted in a public forum on Anatomy Connected, a platform open to all AAA members. Follow‐up interviews were conducted with eight respondents from the pilot survey. Quantitative and qualitative analysis of the survey results, as well as thematic analysis of the interviews revealed that most individuals, regardless of their own completion of a postdoctoral fellowship, see value in a well‐designed anatomy education postdoctoral fellowship experience. Perceived potential benefits include mentorship to enhance their own pedagogy, gain additional teaching experiences, and development of educational research skills. There are concerns regarding the funding support for such a position, as well as the potential exploitation of an individual in such a position. Still, as a whole, the survey and interview results suggest that developing anatomy education postdoctoral fellowships have a strong potential to aid the development of our future anatomy educators and educational researchers, particularly for individuals earning doctorates from programs that do not offer explicit training in pedagogy and/or educational research.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inflammation and Innate Immune Activation During Ex‐Vivo Heart Perfusion
Authors: Qi Xiao , Hatami Sanaz , White Christopher , Himmat Sayed , Aboelnazer Nader , Ondrus Martin , Kinnear Alxe , Wu Yilun , Nagendran Jayan , Freed Darren H.
Published: June 2021

Introduction

Heart transplantation remains the standard treatment for advanced heart failure. However, waiting lists for donor heart continue to rise all over the world due to the discrepancy between the demand and supply of suitable organs. Ex‐vivo heart perfusion (EVHP) has been proposed as a means improving heart preservation and expanding the donor pool. The clinically available EVHP performs in non‐working mode (NWM). Alternatively, EVHP in working mode (WM) may be better in terms of function assessment and preservation.

Objectives

The generation of circulating mediators of inflammation during EVHP has not been previously investigated. We hypothesized that inflammatory mediators will be activated during EVHP and that the inflammatory response would play a role in declining of donor heart function.

Methods

The procured porcine hearts were perfused ex vivo in a beating state for 12 hours (normothermic, whole blood‐based perfusate, no steroids) on a custom EVHP apparatus. Group 1 hearts (n=9) were perfused in a working mode (left atrial pressure=6 mmHg, heart rate=100 beats/minutes) for the entire EVHP interval. Group 2 hearts (n=6) were briefly transitioned into a working mode at hours T1, T5, and T11 for inflammatory and functional assessment, but were otherwise perfused in a non‐working mode (left atrial pressure=0 mmHg). In vivo hearts (n=4) without perfusion were treated as baseline control. Cardiac functional parameters were compared between two perfusion modes groups. The pro‐inflammatory cytokine levels in the perfusate and myocardial tissue were measured by ELISA and western blot and compared between two groups.

Results

Myocardial function declined over time but the function parameters were better preserved in WM (T11 cardiac index (mL/minute/gram): WM=6.9±1.0 vs NWM=2.0±1.2, p=0.02; LV stroke work (mm Hg·mL): WM=1012.5±245.7 vs NWM=303.4±121.6, p=0.03). The perfusate concentration of pro‐inflammatory cytokines TNF‐α, IL‐6, IL‐8, IL‐1α, IL‐1β, IL‐18 increased significantly during the 12‐hour perfusion interval (p<0.05), but were not significantly different between WM and NWM groups. The IL‐6 and TNF‐α increased in the left ventricular tissue after perfusion compared to baseline myocardial samples (p<0.05), with no significant difference between WM and NWM.

Conclusions

A significant pro‐inflammatory cytokine response is generated during prolonged EVHP in the perfusate and left ventricular tissue, independent of perfusion mode. The inflammatory responses may play role in declining of cardiac function. Further studies are warranted to elucidate the stimulants, consequences of inflammatory responses (I.E. on energy metabolism), and methods to mitigate these effects.

Support or Funding Information

Canadian National Transplant Research Program, Canadian Institute of Health Research Grant
Elucidating Epigenetic Readers of H3T45ph in S. cerevisiae
Authors: Grant Patrick , Lee Christina , Pray‐Grant Marilyn , Bekiranov Stefan
Published: June 2021

The phosphorylation of histone H3 threonine 45 (H3T45ph) in S. cerevisiae and mammalian cells occurs at the N‐terminal alpha‐helix and aligns at the precise location of DNA points of entry and exit on the nucleosome. The phosphorylation peaks during S‐phase and is mediated by the DNA replication kinase Cdc7, which is dependent on forming a complex with Dbf4 to phosphorylate the residue. Loss of T45ph leads to greater sensitivity to replicative stress, but it is not required for cell survival. ChIP‐seq was used to identify the genomic sites of H3T45ph in asynchronous and hydroxyurea (HU) treated cells and RT‐qPCR to determine the mark's influence on transcription. These studies revealed that H3T45ph is located at both origins of replication and at the sites of highly transcribed genes responsible for protein synthesis and glycolysis. Here we aim to elucidate the reader(s) of the H3T45ph mark to aid in understanding the possible roles H3T45ph may play in regulating chromatin structure, the recruitment of transcription factors, or the recruitment of the DNA replication or repair machinery. Our research supports the identification of readers of the mark, which perform a wide variety of functions and are classically associated with DNA damage repair pathways and stabilizing stalled replication forks, as well as differentially regulating rapamycin‐mediated transcription. We hypothesize that these factors act as transcription factors that regulate ribosome biogenesis and cell growth, as well as aid in DNA replication. We speculate this pathway is crucial for efficient DNA replication fork stability and the activation of growth genes, coordinating DNA replication and gene expression in highly proliferating cells.

Support or Funding Information

This work is supported by NIH Grant 5R01GM111911.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A structural role for the C‐terminal conserved LSA motifs in Arabidopsis LARP6 proteins
Authors: Toner Chelsea , Chovanec Lindsay , Foster Corina , Foster Jessica , Otte‐Petrill Courtney , Betancourt Francisco , Billey Elodie , Bousquet‐Antonelli Cécile , Lewis Karen
Published: June 2021

La Related Protein 6 (LARP6) is a sub‐family of the RNA‐binding La‐Related Proteins superfamily (LARPs). In vertebrates, LARP6 is known to bind to the 5′ untranslated region of mRNAs that encode for collagen type I. In contrast, the ligands and biological function of the three LARP6 paralogs in plant LARP6 (LARP6a, LARP6b and LARP6c) remain to be determined. Of the three variations of the LARP6 protein in Arabidopsis thaliana, LARP6b and LARP6c contain a PAM2 motif in the N terminal region of the protein, while LARP6a does not. All three LARP6 paralogs contain the conserved LSA motif, which is a distinguishing feature of the LARP6 subfamily and has been implicated in biologically relevant protein‐protein interactions. We have successfully expressed and purified full‐length LARP6a, LARP6b, and LARP6c proteins from Arabidopsis thaliana as recombinant proteins in E. coli. Mutant forms of the Arabidopsis LARP6 proteins were generated using site‐directed mutagenesis to test the role of conserved sequence motifs. The deletion of the PAM2 motif from LARP6c does not negatively impact the structural stability of the full‐length protein. However, deletion of the LSA from LARP6c does appear to destabilize the recombinant protein during biosynthesis. We hypothesize that the LSA motif in plant LARP6 proteins contributes to the global structure and/or the thermodynamic stability of the proteins. These studies using highly purified recombinant protein, with protein stability measured by differential scanning calorimetry, provide novel insight into the potential structure and function of the uncharacterized LSA motif that characterizes the LARP6 subfamily of RNA‐binding proteins.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Toxic effect of resveratrol induced by energy restriction on mitochondrial dysfunction in mouse hepatocytes
Authors: CAMACHO LILLIAN , GOMEZ MINERVA RAMOS , PEREZ LUIS ALBERTO MADRIGAL , LORÍA JORGE LUIS ROSADO
Published: June 2021

Resveratrol (RSV) is a polyphenol classified as a phytoalexin, which occurs naturally and in response to a mechanical injury or biotic and abiotic factors. RSV has been shown to reduce cell viability of pathogenic organisms, probably through inhibition of the electron transport chain (ETC) and F1F0‐ATPase (which is referred to as oxidative phosphorylation in general), generating a decreased synthesis of energy, causing, in turn, a mitochondrial dysfunction. Interest in RSV has increased in recent years due to the beneficial health effects shown by this stilbene. However, such beneficial effects have been observed only at high concentrations of glucose or lipids, without considering those with adequate or low consumption in which the beneficial effect could be neglected and even show toxic effects. Therefore, the objective of the investigation was to determine whether the amount of glucose modulates the effects of RSV on mitochondrial dysfunction in mouse hepatocytes (Hepa1c1c7). Hepa1c1c7 cells were supplemented with both RSV (0.1 to 1000 μM) and glucose (0.5 to 30 mM). According to the MTT assay, at high concentration of RSV (1000 μM) cell viability significantly decreased by 91% at low glucose concentrations (0.5 to 1 mM), while normal glucose concentration (5 mM) decreased cell viability by 63%, relative to control cells. Furthermore, RSV had no effect on cell viability at high glucose concentrations (20 and 30 mM); this indicates that there is a RSV‐dependent relationship with the energetic state of the cell. Subsequently, mitochondrial dysfunction was determined using membrane potential, reactive oxygen species (ROS) production, and ADP/ATP ratio. In the ROS production experiment, a decrease in ROS release was observed at the highest glucose concentrations (10, 20 and 30 mM) in cells supplemented with 10, 100 and 1000 μM of RSV, which indicates that RSV inhibits ROS overproduction in a dose‐dependent manner, demonstrating its antioxidant effect. Interestingly, under energy restriction concentrations (glucose 0.5 and 1 mM) at concentrations greater than 10 μM of RSV, an increase of ROS release was observed, which corroborates the fact that under restraint energy RSV exerts a toxic effect. Besides, an increase in both mitochondrial membrane potential and ADP/ATP ratio was observed; therefore, it was suggested that RSV is interrupting the flow of electrons, altering the CTE, with a subsequent decrease in ATP production. Altogether, these data support the hypothesis that the toxic effects of RSV are dependent on the cellular energy restriction condition as a result of mitochondrial dysfunction.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Feedback Regulation of HMG‐CoA Reductase in Livers of Mice
Authors: Su Shan , Garland Kristina , Jo Youngah , Hwang Seonghwan , Young Gennipher , Fuentes Iris , Schumacher Marc , Elsabrouty Rania , Johnson Brittany , Debose‐Boyd Russell A.
Published: June 2021

Introduction

HMG‐CoA reductase is a membrane protein of the endoplasmic reticulum (ER) that catalyzes the reduction of HMG‐CoA to mevalonate, a rate‐limiting step in the synthesis of cholesterol and nonsterol isoprenoids. Sterol and nonsterol isoprenoids exert stringent feedback control on HMGCR through multiple mechanisms. This ensures constant synthesis of essential nonsterol isoprenoids, while avoiding toxic overaccumulation of cholesterol. One of these mechanisms involves sterol‐induced ubiquitination of HMGCR, which marks the enzyme for degradation from ER membranes that is augmented by nonsterol isoprenoids. In this study, we examine the contribution of this sterol‐accelerated ubiquitination/degradation to overall regulation of HMGCR in the livers of mice.

Methods

Forty mice, including 20 wild‐type (WT) and 20 knock‐in (Ki) mice that express ubiquitination‐resistant HMGCR, were fed diets containing only chow, or chow supplemented with 0.1%, 0.3%, or 1% cholesterol. After five days of feeding, livers were harvested for measurements of cholesterol and triglycerides, immunoblot analysis of six proteins, and qRTPCR of 26 genes related to cholesterol, nonsterol isoprenoid, and fatty acid synthesis.

Results

Normalization of mRNA levels to protein levels indicates that HMGCR Ki mouse livers contain a higher level of HMGCR protein despite mRNA downregulation. Protein and gene expression of SREBP‐2 and its target genes, which contribute to cholesterol synthesis, decreased as expected with increased dietary cholesterol. Conversely, protein and gene expression of SREBP‐1 and its target genes increased, likely due to SREBP‐1c predominance toward fatty acid synthesis, which prevents cholesterol accumulation.

Conclusion

The increase in HMGCR protein relative to mRNA suggests that significant posttranscriptional regulation exists in the form of impaired degradation. Furthermore, these normalized values indicate that accumulation of protein is primarily due to impaired degradation at lower cholesterol levels (chow, 0.1%); however, at high cholesterol levels (0.3%, 1%), a greater degree of transcriptional control from sterol‐mediated inhibition of SREBP‐2 regulates HMGCR due to negative feedback. This study demonstrates the role of degradative control on inhibition of HMGCR and may assist in reducing HMGCR accumulation during statin therapy.

Support or Funding Information

NIH grants HL020948, GM112409, Ruth L. Kirschstein National Research Service Award (NRSA) Short‐Term Institutional Research Training Grant (T‐35)
Pathophysiological and neurobehavioral characteristics of a propionic acid‐mediated autism‐like rat model
Authors: Choi Jeonghyun , Lee Seunghoon , Won Jinyoung , Jin Yunho , Hong Yunkyung , Hur Tai‐Young , Kim Joo‐Heon , Hong Yonggeun
Published: June 2021

Autism spectrum disorder (ASD) is induced by complex hereditary and environmental factors. However, the mechanisms of ASD development are poorly understood. The purpose of this study was to identify the standard indicators of this condition by comparing clinical, pathophysiological, and neurobehavioral features in an autism‐like animal model. A total of 22 male Sprague‐Dawley (SD) rats were randomly divided into control and 500 mg/kg propionic acid (PPA)‐treated groups. The rats were subjected to behavioral testing, gene expression, and histology were examined to detect pathophysiological and neurobehavioral alterations. Exploratory activity and non‐aggressive behavior were significantly reduced in PPA‐treated rats, whereas we observed enhanced aggressive behavior in terms of adjacent interactions on Day 14 after PPA treatment. To evaluate gene expression after PPA administration, we analyzed tissue from the hippocampus using reverse transcription polymerase chain reaction (RT‐PCR). Glial fibrillary acidic protein (GFAP) was augmented in the PPA‐treated group on Day 14 after ASD induction, whereas octamer‐binding transcription factor 4 (OCT4) expression was significantly decreased in the PPA‐treated group. The histological evaluation revealed significantly reduced granule cells diameter and granule cell layer (GCL) thickness in PPA‐treated rats compared with control rats. We suggest that PPA administration induced abnormal neural cell organization, which may have led to autism‐like neurobehaviors including increased in aggressive behavior, reduced in exploratory activity, and isolative and passive behaviors.

Support or Funding Information

Funds: NRF‐2017R1A2A2A01067169, KGM4611714, 2016 Creative Research Program of Inje University, Republic of Korea

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pyrrole‐protein adducts, a mechanism‐based biomarker of pyrrolizidine alkaloid induced hepatotoxicity
Authors: Ma Jiang , Li Mi , Ye Yang , Fu Peter P. , Lin Ge
Published: June 2021

Pyrrolizidine alkaloids (PAs) are among the most hepatotoxic phytotoxins present in about 3% of the flowering plants. Ingestion of PAs causes hepatic sinusoidal obstruction syndrome (HSOS), characterized by primary sinusoidal endothelial cell damage. The hepatic cytochrome P450‐mediated metabolic activation of PAs leading to the formation of pyrrole‐protein adducts has been considered the major pathway in initiating PA‐induced liver injury (PA‐ILI). Our previous studies provided evidence to support that pyrrole‐protein adducts can potentially serve as a biomarker of PA exposure and PA‐ILI. The present study aimed to 1) investigate the correlation between the level of pyrrole‐protein adducts formed from the metabolism of different PAs and their hepatotoxic potency; and 2) characterize the binding specificity, dose‐response, and elimination kinetics of blood pyrrole‐protein adducts in rats. Two PA‐containing herbs Crotalaria sessiliflora (containing senecionine and seneciphylline) and Gynura segetum (containing monocrotaline), and a representative toxic PA (monocrotaline) were used for the study. We observed that there was a good correlation between the level of dose‐dependent hepatotoxicity (determined by the elevation of serum ALT) and the formation of pyrrole‐protein adducts in the liver of rats treated with different PAs. There was also a good correlation between the hepatic and the blood on the level of pyrrole‐protein adduct formation. The results indicated that pyrrole‐protein adducts were formed dose‐dependently in all the monocrotaline‐treated rat blood protein fractions, including hemoglobin (Hb), plasma, albumin, and residual protein fractions, with the highest level of pyrrole‐protein adducts in Hb fraction. The elimination half‐lives of pyrrole‐protein adducts in rat Hb, plasma proteins, albumin, and residual proteins were determined to be 12.08 ± 1.06, 2.81 ± 0.08, 2.54 ± 0.11, and 2.93 ± 0.04 days, respectively. These findings suggest that pyrrole‐Hb adduct is a potential minimal invasive biomarker of PA exposure and PA‐ILI.

Support or Funding Information

The present study is supported by Research Grant Council of Hong Kong (GRF Grant, Project No.: 471013), CUHK One‐off Funding for Joint Lab/Research Collaboration (Project Code: 3132968) and CUHK School of Biomedical Sciences – Seed Fund for Joint Establishments.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enhanced expression of pro‐renin receptor and proinflammatory cytokines in Npr1 gene disrupted mice
Authors: Periyasamy Ramu , Das Subhankar , Pandey Kailash N.
Published: June 2021

Atrial and brain natriuretic peptides (ANP and BNP) activate guanylyl cyclase/natriuretic peptide receptor‐a (GC‐A/NPRA), which regulates blood pressure through inhibition of renin‐angiotensin‐aldosterone system (RAAS). The present study was aimed to determine whether targeted‐disruption of Npr1 gene (coding for GC‐A/NPRA) upregulates pro(renin) receptor (P)RR expression and leads to activation of mitogen activated protein kinases (MAPKs) in Npr1 gene‐knockout mice. The results showed that Npr1 gene disruption enhanced renal (pro)renin receptor expression by 3‐fold in 0‐copy (−/−) mice compared with 2‐copy (+/+) control mice, which in turn was associated with increased expression of angiotensin converting enzyme (ACE) and angiotensin receptor type 1 (AT1R) gene by 2‐fold and 3‐fold respectively. To identify the angiotensin II‐ (Ang II) dependent and ‐independent mechanism of (P)RR, ACE inhibitor captopril and AT1R antagonist losartan were administered to Npr1 mice genotypes. Both drugs did not alter the expression of (pro)renin receptor in all Npr1 mice genotypes. In parallel, renal expression of (MAPKs) p‐Erk1/2 and p‐p‐38 were increased by 2.5‐fold and 3‐fold in 0‐copy mice compared with 2‐copy control mice. Treatment with captopril did not alter the expression of p‐Erk1/2 and p‐p38 in Npr1 mice genotypes compared with their untreated mice counterparts. Interestingly, losartan reduced the expression of p‐ERK1/2 and p‐p38 in 1‐copy mice compared with vehicle‐treated control groups. The present findings suggest that disruption of Npr1 gene upregulates (P)RR and activates MAPKs p‐Erk1/2 and p‐p38 in 0‐copy mice, independent of Ang II generation by enhancing pro‐inflammatory cytokines in a gene dose‐dependent manner.

Support or Funding Information

NIH/NHLBI: HL57531

NIH/NHLBI: HL62147

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of Alpha‐1 Adrenergic Vasoconstriction in Regulating Skeletal Muscle Blood Flow during Single Leg Knee Extension Exercise with Advancing Age
Authors: Theisen Jeremy L. , Ratchford Stephen M. , Clifton Heather L. , Bunsawat Kanokwan , Barret‐O'keefe Zachary , Broxterman Ryan M. , Gifford Jayson R. , Hydren Jay , Rossman Matthew J. , Ives Steve J. , Witman Melissa A.H. , Trinity Joel D. , Garten Ryan J. , Morgan David E. , Nelson Ashley D. , Richardson Russel S. , Wray D Walter
Published: June 2021

Dynamic exercise evokes an increase in sympathetic nervous system (SNS) activity, a response that contributes to the “fine‐tuning” of blood flow to meet the metabolic demands of active skeletal muscle. Expression of this exercise‐induced increase in SNS activity is blunted within the vasculature of exercising muscle, an event known as “functional sympatholysis”. However, how this process may be altered with advancing age remains unclear. Therefore, the purpose of this study was to determine the role of alpha‐1 adrenergic vasoconstriction in the regulation of exercising skeletal muscle blood flow and the modulating effect of age. Eleven young (6 male, 27±1 yrs) and six older (6 male, 60±4 yrs) healthy volunteers participated in this study. Leg blood flow (LBF; ultrasound Doppler), mean arterial blood pressure (MAP), and leg vascular conductance (LVC) were evaluated at rest and during dynamic single‐leg knee‐extension exercise (0, 5, 10 and 15 W) before and after blood flow‐adjusted intra‐arterial administration of the alpha‐1 adrenergic agonist phenylephrine (PE, 0.03 ug·ml−1). At rest, there was no difference between young and older subjects in the PE‐induced decrease in LBF (−37±4% vs. −31±3%, young vs. older) or LVC (−53±6% vs. −48±2%, young vs. older). During exercise, the PE‐induced decrease in LBF was attenuated in an intensity‐dependent manner, but was not different between young (0W: −20±4%; 5W: −16±3%; 10W: −14±4%; 15W: −5±3%) and older (0W: −15±4%; 5W: −9±4%; 10W: −10±2%; 15W: −5±4%) subjects. Similarly, the PE‐induced decrease in LVC was attenuated as work rate increased, but was not different between groups at any work rate. MAP increased with exercise, but did not differ between groups or as a result of drug administration. Alpha‐1 adrenergic vasoconstriction was similar between young and older adults at rest and during low to moderate intensity knee‐extension exercise, suggesting that functional sympatholysis is not altered with advancing age.

Support or Funding Information

This project is funded in part by the National Institutes of Health (HL118313) and the U.S. Department of Veterans Affairs (RX001697, RX001418, E6910R).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Morphological Changes to the Cruciate Ligaments and Menisci in Relation to ageing.
Authors: Hirtler Lena , Toader Ian‐Philip , Röhrich Sebastian , Kainberger Franz
Published: June 2021

In this retrospective, cross‐sectional MRI based study on the anatomy of the cruciate ligaments (ACL, PCL) and the menisci (MM, LM) of the human knee, the goal was to identify possible age‐ and gender dependent variations in morphometry of these structures.

332 patients were included, 184 women (55,4 %) and 148 men (44,6 %), all who underwent an MRI examination of the knee. Age distribution ranged from 1 year (y) to 78 y with a mean of 33 years. All patients were subdivided in 6 age groups (younger then 10y, 11–20y, 21–30y, 31–45y, 45–55y, older then 56y). Exclusion criteria consisted of poor image quality as well as destructive and space occupying, pathological processes in close vicinity to the capsular apparatus and investigated structures, respectively.

Measurements included following parameters. ACL: length, blumensaat angle, ACL‐tibia angle, cross‐sectional‐area (CSA), diameter (APD), width, intercondylar notch width (ICN); PCL: length, PCL angle, Angle Ratio, CSA, APD, width; Menisci: height of the anterior and posterior horns, height and width of the mid‐body, axial APD of the menisci, tibia width.

There were significant correlations between age and length, CSA, APD, width and both angles of the cruciate ligaments. Male ACL size was greater in nearly every parameter when compared to the female population. Merely Blumensaat and tibia angle showed greater values in the female cohort. ACL/ICN‐ratio also correlated significantly with age. Especially when evaluating the younger age groups significant differences could be observed. Sagittal height of the meniscus horns correlated significantly with coronal height of the meniscus body in several age groups. There was also a significant correlation between medial and lateral meniscus height. Tibia width presented a significant but weak predictor of medial and lateral meniscus width. This is one of the largest studies evaluating the cruciate ligaments and menisci in different phases of life when considering cohort size. The results concur with most of the results stated in previous and similar studies.

Although there has been an abundance in studies about the cruciate ligaments and the menisci in the last decades there is still little detail known about the morphological changes of these structures in different phases of life. The results of this study aim to increase the anatomical knowledge of the investigated structures as well as to establish standard measurements for further studies and to differentiate between normal and pathologic conditions.

Support or Funding Information

No funding was received.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The LexA‐regulated gene ybfE plays a role in DNA metabolism in E. coli
Authors: Hotchkiss Amy Blondin , Kramer Caitlin , Beuning Penny
Published: June 2021

DNA is constantly exposed to damage‐causing agents. The SOS DNA damage response in Escherichia coli is regulated by the LexA protein, and controls the expression of more than 50 different genes. One of the many genes regulated by the SOS response encodes the uncharacterized protein YbfE. Previous work in our lab has shown ybfE is essential to cell survival after exposure to alkylating agents. In order to characterize ybfE further, two open reading frames are being examined because two different in vitro transcription start sites have been proposed. One open reading frame encodes an additional 23 amino acids at the N‐terminus of the protein and has been shown to cause a striking decrease in cell survival upon overexpression. To Identify genetic interactions, a series of knockout strains have been investigated to determine whether deletion of any one gene alleviates the lethality associated with overexpression of ybfE. To date assays in strains harboring deletions of various DNA repair proteins have not suppressed the lethal overexpression phenotype. In addition, hydroxyurea sensitivity studies are being conducted. Hydroxyurea is known to inhibit DNA replication without inducing DNA damage, allowing further investigation into the role ybfE plays in the SOS response. Deletion of the ybfE gene reduces the susceptibility of wild‐type E. coli to HU suggesting a role for YbfE in DNA metabolism. Further investigation into protein interactions and the role of YbfE in the SOS response are ongoing.

Support or Funding Information

Support from NSF‐MCB‐1615946 and American Cancer Society RSG‐12‐161‐01‐DMC.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.