bluuebunny/crossref_metadata_2025 huggingface dataset:
The Secretory Profile of Inflammatory Mediators is Altered in Differentiated Primary COPD Samples at Air Liquid Interface Compared to Healthy Equivalents
Authors: Kelly Gillian A. , Douglas Lisa E.J. , Maye Jessica R. , Reihill James A. , McGarvey Lorcan P. , Lundy Fionnuala T. , Litherland Gary , Martin S. Lorraine
Published: June 2021

In the UK, over 1 million people live with diagnosed chronic obstructive pulmonary disease (COPD) however, it is estimated that this is only 1/3 of the total COPD population. Diagnosis is based on patient airflow obstruction, a late developing symptom, by which time a significant amount of lung damage has already occurred (1). Investigation into a range of biomarkers to aid earlier identification of COPD is therefore essential.

Apical and basolateral samples collected from differentiated primary healthy and COPD cells at air liquid interface (ALI) were screened for 35 proteases, 32 protease inhibitors and 37 other inflammatory mediators using R&D Systems Proteome Profiler Protease array and Anti‐Protease array and BioRad Bio‐Plex Pro Human Inflammation Panel 1, 37‐Plex Assay respectively, with the aim of identifying potential biomarkers.

Both apical and basolateral secretory profiles were altered in COPD samples compared to their healthy counterparts. The majority of apically secreted proteases were up‐regulated whereas, protease inhibitors were down‐regulated. Basolateral secretion of inflammatory mediators was found to be both up‐ and down‐regulated depending on which mediator was being measured.

These initial results highlight interesting targets which will be further investigated in clinical samples to assess their potential as biomarkers to aid the earlier diagnosis of COPD.

Support or Funding Information

BREATH (Border and REgions Airways Training Hub) is funded by the EU's INTERREG Va programme managed by the Special EU Programmes Body (SEUPB).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of muscle‐derived BAIBA on osteocytes with aging
Authors: Bonewald Lynda F.
Published: June 2021

Beta‐aminoisobutyric acid, BAIBA, is a metabolite released into the circulation by contracted muscle. It has been shown to have beneficial effects on metabolism, glucose tolerance, the browning of white fat, and recently on osteocyte viability and function. BAIBA prevents both bone and muscle loss due to unloading and synergizes with anabolic loading to promote bone formation. Combining BAIBA with voluntary wheel running of 12 mo old female mice for 6 mo until 18 mo of age preserved the capacity of the extensor digitorum longus and soleus muscles to produce factors that maintain osteocyte viability. This capacity is normally lost with aging. Ex vivo contraction of isolated murine muscle from both young (5 mo) and old (22mo) mice induces secretion of BAIBA, showing that with age muscle can still produce this factor with contraction. However, with aging, osteocytes appear to lose expression of the Mas related G protein receptor type D, the receptor in osteocytes that responds to BAIBA. Efforts are underway to identify means to either maintain or overexpress this receptor with aging.

Support or Funding Information

NIH NIA PO1AG039355

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How the Formation of the Turing Pattern Could Be Used in a Bio‐Mechano‐Chemical Framework for Describing Development of Bones and Joints
Authors: Alvarado Diego Alexander Garzón
Published: June 2021

In this conference, we will show our mathematical models for interpreting different biological, biomechanical, and biochemical processes that are fundamental on bone and joints formation. In order to do this description, we use a set of reaction‐diffusion equations that are widely used in morphogenesis, in which biochemical factors are assumed to be secreted by precursor cells, mesenchymal cells, chondrocytes, and osteoblasts, in endochondral and intramembranous ossification, respectively. The solution leads to the so‐called Turing patterns, which represent these processes of ossification approximately. Besides, it has been included the mechanical effects on the growth and form in the development of joints, primary and secondary ossification centers, and bones. Those models have the property of describing the development and shape of bones and joints accurately. In this presentation, we will show the model and hypotheses used for describing the development of the skull, clavicle, patella, and finger joints. Also, we will present some of the research that we are doing around experimental and mathematical problems in biomedical engineering at Universidad Nacional de Colombia.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mind the Gap: How Reference Samples Impact Skeletal Age Estimation in Forensic Casework
Authors: Langley Natalie R.
Published: June 2021

Forensic anthropologists are often tasked with determining skeletal age‐at‐death to assist law enforcement in identifying skeletal remains. If the remains are skeletally mature, degenerative changes in articular surfaces are typically assessed to determine age. If the skeleton is immature tooth eruption, dental development, and/or epiphyseal union are used to estimate age. Forensic anthropologists derive an age‐at‐death estimate by comparing our assessment of skeletal age indicator(s) to standards derived from skeletal collections of known age‐at‐death. Prior to the establishment of modern skeletal collections such at the William M. Bass Donated Collection at the University of Tennessee, anthropologists developed age estimation techniques using late 19th and early 20th century anatomical collections (e.g. the Hamann‐Todd Osteology Collection at the Cleveland Museum of Natural History and Terry Anatomical Collection at the Smithsonian Institution). However, research on Bass Collection skeletons has demonstrated remarkable changes in skeletal morphology in modern Americans compared to early 19th century birth cohorts, illustrating that standards derived from modern populations are imperative for forensic casework; anatomical collections and archaeological samples are not appropriate reference samples. Age estimation in forensic casework is further complicated by the age distribution of the forensic case population. Forensic cases often involve accidental deaths, homicides, and suicides of individuals from a younger age demographic than natural deaths which comprise many of the reference samples used to derive age estimates. This difference between the age distribution of the reference sample and that of the “target” population leads to ‘age mimicry’, whereby skeletal age‐at‐death may be under‐estimated. This presentation will highlight research that illustrates the impact of secular change and reference sample composition on the accuracy and precision of forensic skeletal age‐at‐death estimation. The documentation of significant changes in skeletal morphology illustrates what evolutionary biologists already know—phenotype is dynamic, not static. These short‐term evolutionary changes in skeletal form have implications for anatomical research beyond the hard tissues.

Support or Funding Information

National Institute of Justice

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CRISPR‐based functional evaluation of common SZ risk variants
Authors: Schrode Nadine , Ho Seok‐Man , Brennand Kristen
Published: June 2021

Schizophrenia (SZ) is a highly heritable neuropsychiatric disorder. Nevertheless, the functional mechanisms through which the large contribution of common genetic variation underlies this disorder remain unclear. GWAS and eQTL analyses have previously identified candidate genes, whose expression is affected by common SZ risk variants (SZ‐eQTL genes). We established a genetics‐driven hiPSC‐based approach for the prioritization and functional validation of common variants and genes associated with SZ.

Using this strategy, we prioritized one putative causal SZ‐eQTL SNP and five SZ‐eQTL genes for functional evaluation. By integrating CRISPR‐mediated gene editing, activation and repression technologies, our isogenic hiPSC‐based neuronal platform recapitulated genotype‐dependent gene expression differences from post‐mortem brains in SNP‐edited hiPSC‐neurons, identified evidence of molecular convergence downstream of common variant perturbations and uncovered specific pre‐ and post‐synaptic neuronal deficits associated with the SZ‐eQTL gene SNAP91.

We demonstrate a systematic and comprehensive strategy to interpret and evaluate the growing number of SZ‐associated variants and genes across neural cell types and genetic backgrounds, which will be applicable to other complex genetic disorders driven in large part by common variation.

Ultimately, our objective is to understand the cell‐type specific contributions of SZ risk variants to disease predisposition.

Support or Funding Information

This work was partially supported by National Institute of Health (NIH) grants R01 MH101454 (K.J.B.), R01 MH106056 (K.J.B.) and R01 MH109897 (P.S. and K.J.B.)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AP‐2α and AP‐2β Cooperatively Regulate Gene Expression of Ectodermal WNT Ligands to Drive Craniofacial Patterning
Authors: Van Otterloo Eric , Milanda Isaac , Li Hong , Williams Trevor
Published: June 2021

Tissue‐tissue interactions are central to proper embryogenesis and are often disrupted in developmental disorders—providing an impetus to further understand their regulation. These interactions require coordinated molecular crosstalk (i.e., signaling molecules) between tissues, thus, providing the instructional cues necessary to generate complex anatomical structures. A prototypical example includes epithelial:mesenchymal interactions—necessary to pattern the craniofacial complex as well as central to the development of a variety of ectodermal appendages. While a great deal of information exists on both the molecules involved in crosstalk and their immediate transcriptional response in adjacent tissues, much less is known regarding how such signals are established. Here, through the use of conditional loss‐of‐function models of AP‐2 transcription factors (AP‐2α and AP‐2β) in mice, we reveal an essential, cooperative role for AP‐2α/β in driving expression of the genes encoding a variety of WNT ligands. Focusing on craniofacial development, loss of AP‐2α/β from the ectoderm results in major dysregulation of the WNT‐signaling pathway, dysregulation of downstream WNT target genes in adjacent tissues, and finally a profound impact on facial form. Importantly, genetic reduction of a WNT‐pathway inhibitor in AP‐2α/β ectoderm mutants can partially mitigate some of these defects. In addition, highlighting a broader role for AP‐2 in regulating WNT signaling, we find numerous WNT‐dependent ectodermal appendages are disrupted in AP‐2α/β ectoderm mutants. Ongoing work is being conducted to identify the molecular nature by which AP‐2 regulates Wnt ligand expression in the face as well as additional structures. Given WNT‐signaling components are often mutated in a variety of human craniofacial disorders and are key components of vertebrate facial evolution, these findings suggest that ectodermal AP‐2 regulation of WNT‐signaling is likely a key axis in these processes.

Support or Funding Information

EVO NIH NIDCR K99 DE026823

TW NIH NIDCR 2R01 DE012728 and 1R01 DE019843

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anatomy as a Science Worth Seeing: Killing Many Birds of Our Academic Role with One Stone
Authors: Chapman Jamie
Published: June 2021

As anatomy academics, formal parts of our role are to engage the community, provide service to our university and to our discipline of anatomy. With increasing demands and workloads, it can be difficult to fulfill all of these different roles. However, it is possible to meet many of these different roles at the same time through anatomy outreach.

To many, the general public and others in the scientific community, anatomy is often seen as a dead science (or simply a science of studying the dead). Our role, in providing service to the discipline of anatomy, is to dispel this misconception and demonstrate that anatomy is very much for the living. As anatomists, we're also lucky in that, compared with many of the other sciences, anatomy has the added bonus of being highly visual – something which is also highly coveted by people interested in marketing and the media. Now, while we cannot typically use cadaveric anatomy to engage the public, there are a number of other activities which we can use – anatomical body painting, for example.

Anatomical body painting (ABP) was introduced to our level 1 introductory Human Biology unit as a means to increase student engagement with the musculoskeletal system. While ABP is not new to anatomy education, it was new to this specific context and to this institution. Through the engagement of the university's marketing team, local media were invited to film the practical and perform interviews. Soon after, cascading invitations to bring ABP to public engagements began to flow and the level of anatomy engagement with the community increased.

ABP was used at the university's Open Days. With the help of a life‐model and a number of volunteer students, over the course of five hours, Anatomy‐Man was painted. Also introduced was the ABP of the hand skeleton on the general public, an anatomical‐themed variation of face‐painting often seen at fairs and birthday parties, which proved extremely popular with younger audiences.

Anatomy was subsequently included in “Science Worth Seeing”, a public science engagement event supported by the Federal Government's Inspiring Australia grant. Anatomy was promoted through ABP, ABP of hand skeletons for children and the use of plastic anatomical models as jigsaws, all of which enabled multiple meaningful interactions and discussions about the human body with children and the general public. ABP also made an appearance at a pop‐up science bar during Australia's National Science Week, which featured on a national news bulletin.

Embracing the power of the visual nature of anatomy through activities such as ABP, anatomists can provide service to the discipline, to their institutions and provide outreach to the community.

Support or Funding Information

Partly funded through The University of Tasmania's School of Medicine Staff Research Conference Support.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pharmacogenomics in Indigenous Populations
Authors: Claw Katrina G. , Henderson Lindsay M. , Burke Wylie , Thummel Kenneth E.
Published: June 2021

The integration of genomic technology into healthcare settings has the potential to transform health care through increased personalization of medical decisions. In particular, pharmacogenomic research on drug disposition and response can tailor and improve medication regimens for all patients by informing tests of function altering variation in drug metabolism and transport genes. Unfortunately, Indigenous peoples remain underrepresented in genomic and health research despite having a higher prevalence of heart disease, cancer, diabetes and stroke than any other population. Effective strategies to create research partnerships between tribal communities and genomic researchers are often lacking, yet such partnerships are needed for trustworthy research. As such, Indigenous populations are less likely to benefit from personalized medicine if they are not represented in studies that ascertain genotype‐phenotype relationships that can lead to improved drug treatment efficacy.

We review what is currently known about pharmacogenetic variation in Indigenous populations and evaluate three case studies in which pharmacogenetic research with Indigenous populations were conducted. We highlight the importance of including Indigenous populations in future pharmacogenetic studies and discuss strategies for ethical engagement and collaboration in genomic research with Indigenous communities. The inclusion of Indigenous and other underrepresented communities in genomic research has the potential to expand our understanding of genomic influences on health and improve clinical approaches for all populations.

Support or Funding Information

NIGMS F32 GM119237

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dynamic Control of Gene Expression during mRNA Export and Translation
Authors: Wente Susan R. , Dawson T. Renee , Adams Rebecca L. , Galoda Aditi , Glass Laura , Mason Aaron C. , Sharma Manisha
Published: June 2021

Messenger (m)RNA‐binding proteins (RBPs) are key arbiters in coordinating gene expression from transcription through translation. The specific RBP complement is repeatedly remodeled across the mRNA lifecycle through the activation of distinct RNA‐dependent DEAD‐box ATPases (Dbps in yeast/DDXs in humans). In this manner, the dynamic interactions of mRNA‐RBP complexes (mRNPs) with cellular machinery are controlled in a spatial and temporal manner to direct an mRNA's progression and ultimate fate. For mRNA export through the nuclear pore complex (NPC), an mRNP at the nuclear face must contain the proper complement of adaptor proteins and export receptor to initiate and mediate export, and the removal of key proteins by remodeling at the cytoplasmic NPC face is critical for export directionality. This mRNP remodeling is achieved through localized activation of human DDX19B (S. cerevisiae Dbp5) by Gle1, a multifunctional regulator of distinct Dbps/DDXs, and inositol hexakisphosphate (IP6). At least two Gle1 isoforms are expressed in human cells, with Gle1B required at the NPC and Gle1A in the cytoplasm. We recently uncovered the role for the Nup42 carboxy‐terminal domain (CTD) in mRNA export during the stress response. Altering the minimal Nup42 binding site on Gle1B revealed that mRNA export in HeLa cells requires the Nup42‐Gle1B interaction. In vitro assays with purified, recombinant proteins for both yeast and human systems demonstrated that the Nup42 CTD enhanced the Gle1/Gle1B‐stimulated, RNA‐dependent Dbp5/DDX19B ATPase activity independently of IP6. Of note, the Gle1B‐IP6 stimulatory effect also required purification of DDX19B from a baculovirus insect cell expression system. In contrast to Gle1B at the NPC, Gle1A lacks the Nup42 binding domain, is primarily cytoplasm at steady state, and is not capable of supporting mRNA export. We found that Gle1A is required for translation activity and stress granule dynamics. Furthermore, ERK1/2, JNK and GSK3 targeted Gle1A for stress‐induced sequential phosphorylation. Analysis of gle1A mutants mimicking phospho‐deficient or hyperphosphorylated states revealed roles for its phosphorylation in stress granule dynamics, Gle1 oligomerization, and Gle1‐mediated stimulation of DDX3 activity. Overall, multifactorial control of Gle1 functions by Nup42, IP6, and phosphorylation allows for regulation of gene expression from mRNA export through translation, and provides potential new insights into gle1 dysfunction underlying disease pathologies.

Support or Funding Information

NIH 5R37 GM051219

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Mass Spectrometry in the Advancement of HLA Epitope Prediction
Authors: Abelin Jennifer G.
Published: June 2021

A challenge in developing personalized cancer immunotherapies is the prediction of putative cancer‐specific antigens. Currently, predictive algorithms are used to infer binding of peptides to human leukocyte antigen (HLA) heterodimers to aid in the selection of putative epitope targets. One drawback of current epitope prediction algorithms is that they are trained on datasets containing biochemical HLA‐peptide binding data that may not completely capture the rules associated with endogenous processing and presentation. The field of MS has made great improvements in instrumentation speed and sensitivity, chromatographic resolution, and proteogenomic database search strategies to facilitate the identification of HLA‐ligands from a variety of cell types and tumor tissues. As such, these advances have enabled MS profiling of HLA‐binding peptides to be a tractable, orthogonal approach to lower throughput biochemical assays for generating comprehensive datasets to train epitope prediction algorithms. Today, we will highlight recent progress made in the field of HLA‐ligand profiling enabled by MS and its impact on epitope prediction strategies.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Small HDL, diabetes, and proinflammatory effects in macrophages
Authors: Kothari Vishal , He Yi , Kramer Farah , Barnhart Shelley , Kanter Jenny E , Tang Jingjing , Frey Jeremy M , Vaisar Tomas , Heinecke Jay W , Bornfeldt Karin E
Published: June 2021

Atherosclerosis, an inflammatory disease of the arterial wall, is the commonest cause of cardiovascular disease (CVD) events. Elevated levels of high‐density lipoprotein (HDL) cholesterol are generally believed to protect against CVD and to reduce inflammatory activation of macrophages, a key cell type in lesions of atherosclerosis. However, HDL can also exert proinflammatory effects in macrophages by inducing cholesterol depletion. Because small HDL (7–8.5 nm diameter) is most effective in removing cholesterol from cells by the ABCA1 pathway, we hypothesized that small HDL is the most proinflammatory HDL population. Small‐, medium‐ and large HDL populations were isolated from a human apolipoprotein A‐I transgenic LDL receptor‐deficient mouse model (APOA1Tg;Ldlr−/−). Depletion of cholesterol by HDL increased the ability of toll‐like receptor (TLR) ligands to induce proinflammatory mediators in macrophages. This effect was prevented by cholesterol loading of the macrophages. Small HDL had a significantly stronger effect on Tnfa (3‐fold over LPS alone) and Il1b (2.5‐fold over LPS alone) gene expression than medium or large HDL particles. Inhibition or deletion of the sheddase ADAM17 in macrophages prevented the proinflammatory effects of HDL. Patients with type 1 diabetes mellitus (T1DM) exhibit an increased risk of CVD despite having normal or high HDL‐cholesterol. To investigate the effect of diabetes on HDL subpopulations, we crossed a virally‐induced mouse model of T1DM (Ldlr−/−;GpTg) with the human APOA1Tg;Ldlr−/− mouse. The mice (APOA1Tg;Ldlr−/−;GpTg, n=8–10) were injected with a liver‐specific adeno‐associated virus driving expression of human cholesteryl ester transfer protein (hCETP) to mimic human physiology. Diabetic mice had an increased HDL particle concentration (~2‐fold) associated with a significant increase in serum HDL cholesterol efflux capacity through both ABCA1 and ABCG1, compared with non‐diabetic mice. The largest increase was in the small HDL particle concentration (15 μM vs 6 μM in non‐diabetic mice). Moreover, resident peritoneal macrophages from diabetic mice exhibited increased cytokine gene expression, as compared with non‐diabetic littermates. Our findings demonstrate that higher levels of small HDL, as observed in a mouse model of diabetes, can exacerbate inflammatory activation in non‐lipid loaded macrophages in an ADAM17‐dependent manner. We have found similar changes in HDL concentration and function in humans with T1DM, suggesting that our mouse model might provide novel insights into CVD risk in T1DM patients.

Support or Funding Information

funded in part by the National Institutes of Health, grants DP3DK108209, P01HL092969, P30 DK017047, and P01HL128203

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biomarkers of Hemostatic Dysregulation and Inflammation in Lymphoma: Potential Relevance to Thrombogenesis
Authors: Bontekoe Emily , Antic Darko , Jocik Vera , Hoppensteadt Debra , Siddiqui Fakiha , Baig Nausheen , Fareed Jawed
Published: June 2021

Objective

Cancer‐associated thrombosis and venous thromboembolism (VTE) continue to be a major complication of malignancy. There is increased incidence of clinically overt deep thrombosis (DVT) in cancer patients. Profiling biomarkers involved in hemostatic dysregulation and inflammatory processes in the lymphoma patient population may provide additional insight into the pathogenesis of thrombotic complications in lymphoma. This study aimed to profile hemostatic dysregulation and inflammatory biomarkers in patients with Hodgkin lymphoma, non‐Hodgkin lymphoma and chronic lymphocytic leukemia/small lymphocytic lymphoma. The primary hypothesis of this study is that hemostatic activation and inflammatory biomarkers may be increased in lymphoma patients.

Methods

Citrated blood samples were collected from the Hematology Clinic, University of Belgrade, Serbia. Plasma levels of plasmin activator (PAI‐1), D‐dimer (DD), Factor XIIIa (XIIIa), C‐reactive protein (CRP), microparticle (MP), Von Willebrand Factor (vWF), Protein S (PS), urokinase like plasminogen activator (uPA), tumor necrosis factor alpha (TNF‐α), Beta 2 glycoprotein (β2GPI), and Fibronectin (FN) were measured in 96 lymphoma patients, along with a control/pooled plasma prepared from 50 healthy normal individuals. Commercially available ELISA kits were used for profiling these markers. All results were compiled and calculated as mean + SD and % change.

Results

The results from this study demonstrated elevated levels of DD, CRP, vWF, TNF‐α, uPA, and PS in comparison to normal human plasma. Plasma levels were decreased for PAI‐1 (3%), XIIIa (5%), MP (46%), β2GPI (8%) and FC (15%). Of the lymphoma patient samples with elevated levels of biomarkers, 34% of individual patients had elevated DD, 66% had elevated CRP levels, 50% had elevated vWF levels, 80% had elevated levels of TNFα, and 90% had elevated levels of uPA. However, when comparing each specific type of lymphoma in relation to the ratio of each biomarkers (DD, CRP, vWF, TNF‐α, and uPA), there was no significant difference between individual lymphoma subgroups (p>.05).

Conclusion

The elevated levels of hemostatic dysregulation and inflammatory biomarkers in lymphoma patient samples are suggestive of an increased VTE risk in these patients. The elevated DD, CRP, vWF, TNFα, and uPA clearly suggest a trend towards increased thrombogenesis. Individual patients showing increased amounts of each elevated biomarker proves the significance and use of D‐dimer, CRP, vWF, TNFα and uPA in identifying the level of thrombosis risk.

Support or Funding Information

None.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How comparative anatomy provides insight into human anatomy
Authors: Throckmorton Zachary
Published: June 2021

Human anatomy instructors in the United States have remarkably varied backgrounds for an academic discipline; we work alongside those trained at every scale of biological organization, from molecular genetics to ecology, and these different training regimens provide anatomy departments with a variety of strengths. In this talk, I outline and detail how comparative whole organism anatomy can be useful in teaching human anatomy, primarily in the context of traditional lectures (at both the undergraduate and graduate levels) through three broad categories. First, understanding how relatively anatomically more complex animals like humans develop embryologically is facilitated by appreciating the anatomy of relatively simpler animals like platyhelminth and annelid worms; furthermore, some developmental defects are better understood through the lens of evo‐devo (i.e., normal conditions in other animals are sometimes abnormal conditions in humans, like syndactyly in bats). Second, comparative functional biomechanics yields insight into normal and abnormal musculoskeletal function. The trade‐offs between joint stability or flexibility, or power arm ratios for power or speed, are well‐illustrated by extreme morphology of homologous structures in other animals, such as the incredibly speedy ankles of cheetahs compared to the relatively powerful ankles of humans. Third, much of any scientific professional's development relies upon mastery of Greek‐ and Latin‐derived vocabulary; genus and species names of animals can help anatomy students better remember structural terms. For example, Pterodactylus was a remarkable genus of flying reptiles, and understanding the etymology of the Greek words “pteron” (wing) and “dactyl” (finger) helps students appreciate how the sphenoid bone's pterygoid process and the pterygoid muscles are wing‐shaped, and the clinical conditions syndactyly and polydactyly refer to webbed or conjoined fingers and extra fingers, respectively.

Application of human anatomy knowledge by physicians to animals gave rise to the field of comparative anatomy in the 1600s; Edward Tyson, a human physician, recognized chimpanzees were more like humans than monkeys, and porpoises were more like land mammals than fish from detailed comparative gross dissections. These observations of patterns in the anatomy of humans and animals contributed to Darwin and Wallace independently developing the theory of evolution, which underpins the national One Health initiative. Recognizing humanity's shared ancestry with animals ensures a measure of shared descendance in the instruction of human and animal anatomical education.

Support or Funding Information

This work was supported by the American Association of Anatomists and the Arkansas College of Osteopathic Medicine.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dopaminergic Circuits in Reward and Aversion
Authors: Lammel Stephan
Published: June 2021

The mesocorticolimbic dopamine (DA) system, which is composed of DA neurons in the ventral tegmental area (VTA) projecting to nucleus accumbens and medial prefrontal cortex, has been intensively studied because of its importance in reward processing, drug addiction and other neuropsychiatric diseases. Importantly, while VTA DA neurons were thought to form a homogeneous cell population, recent research has demonstrated a much greater diversity of VTA DA cell type and function. Accordingly, VTA DA neurons encode much more than reward and also contribute to negative affect such as aversion and social defeat stress. How VTA DA neurons could mediate both reward and aversion is currently unknown and an important goal of my research. In my presentation, I will discuss work from the lab that has elucidated the circuit architecture and function of the reciprocal connectivity in the mesolimbic DA system (Yang et al., 2018; Neuron). Moreover, I will discuss recent data that demonstrate the identification of a subcircuit within the mesolimbic DA system that contributes to the encoding of conditioned and unconditioned aversive stimuli (de Jong et al., Neuron, in press). Collectively, our work suggest that we need to develop a new perspective on the DA circuitry that will guide future treatment strategies for addiction and other neuropsychiatric disorders where dysfunction of the neural systems underlying motivated behaviors have been strongly implicated.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SK2/3 modulation of neural development
Authors: Ogundele Olalekan M
Published: June 2021

Regulation of ionotropic NMDA Receptor (NMDAR) potentiation at hippocampal synapses is dependent ‐ partly ‐ on ion channel gating by Ca++ activated K channels (SK2/3). Given that NMDAR is involved in the regulation of neuronal migration and synaptic development, the objective of this study is to determine whether an increased activity of SK2/3 during a critical period of brain development can alter neural circuit formation.

A single dose of intrahippocampal injection of CyPPA (SK2/3 potentiator) was stereotactically injected into the hippocampus and cortex of post‐natal day (PND) 2 mice [SK2/3(+)]. At PND55, the brain of the treated animals and sham were microdissected and processed for immunofluorescence (Saggital) and immunoblot analysis.

Here, we showed that pharmacological hyperactivation of SK2/3 in the neonatal brain significantly ablated the expression of synaptic kinases that are associated with NMDAR signaling in development, and adult synaptic function. Notably, CaMKIIα, PI3K, and PKB expressions were significantly downregulated in the hippocampus of SK2(+) mice when assessed at PND55. These changes were associated with altered neuronal distribution and path of migrating neural projections in the hippocampus and cortex of mice. Aberrant migration of lower layer cortical neurons (Tbr1+) neurons was evident in the layers of the cortex and the CA1 of the hippocampus. The distribution of synaptic structural proteins ‐ Homer1 and F‐actin – further underscores the re‐distribution of neural projections and synapses in SK2/3(+) brain.

Support or Funding Information

LSU SVM Faculty Start up, LSU SVM Corp Grant

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Circadian Control of Animal Physiology
Authors: Sehgal Amita , Zhang Shirley , Artiushin Greg
Published: June 2021

We are interested in the mechanisms that generate circadian (~24 hour) cycles of physiology and behavior. Our studies use primarily a Drosophila model, which was invaluable for dissecting molecular mechanisms of the clock, but they also include mammalian models where appropriate. We are interested in the control of behavior by brain clocks and in the control of physiology by brain and peripheral clocks. We recently identified a clock in the Drosophila blood brain barrier (BBB) that confers a 24 hour rhythm on to permeability of the BBB. Specifically, the BBB clock regulates the activity of efflux transporters, such that these transporters pump xenobiotics out of the brain during the day, but are less active at night, thereby increasing BBB permeability at night. The underlying mechanism involves rhythmic expression of gap junctions that serve to maintain 24 hour cycles of magnesium, a positive regulator of efflux transporters, in BBB cells. As many drugs are targets of efflux transporters, we asked if permeability of such drugs is different at different times of day. Indeed, we found that phenytoin/Dilantin has time‐of‐day specific effects in a drosophila seizure model. In recent work, we have characterized rhythms of the BBB in mammalian cells and are addressing the relevant mechanisms.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How Replication Stress Drives Genome Instability
Authors: Forsburg Susan
Published: June 2021

DNA replication stress can result from intrinsic challenges (such as defective replication proteins, collisions with transcription machinery, or synthesis through repetitive sequences), or extrinsic agents (such as agents that damage DNA (UV, MMS, CPT) or starve for nucleotides (HU)). The cell deploys a variety of checkpoint and repair proteins to manage accumulation of single stranded DNA, replication fork stalling, fork regression and recombination intermediates to repair. Increasingly, it is clear that the response to stress is not just important for completion of S phase, but impacts mitosis and faithful chromosome segregation. Thus, response to replication stress is an initial barrier to malignant transformation.

Our study employs a model genetic system, the fission yeast S. pombe. We are using mutations that attenuate core components of the replication machinery or epigenetic components, to investigate different kinds of stress and how they engage, or evade, the normal cellular response. Our approach is grounded in analysis of the intact cell and determining the consequences of replication stress including mutation, chromosome mis‐segregation, or gross chromosome rearrangements and translocation. A key component is monitoring stress responders in real time using dynamic live cell imaging. Recent studies have focused on how checkpoint evasion leads to ongoing mitosis and chromosome rearrangement, and on how the response of cells in the meiotic pathway differs from the vegetative cell cycle. Current investigations will be discussed.

Support or Funding Information

NIH R35 GM118109

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PMNs promote Non‐Homologous End‐Joining (NHEJ)‐mediated survival of colorectal cancer cells in inflammatory setting
Authors: Bui Triet M , Wiesolek Hannah , Butin‐Israeli Veronika , Sumagin Ronen
Published: June 2021

Exacerbated Inflammation that can be driven by neutrophil (PMN) infiltration is one of the major risk factors for development and progression of colorectal cancer (CRC). Using a biopsy‐based acute‐injury model, we previously showed that miRNAs derived from PMN‐microparticles (MPs) inhibit Homologous Recombination (HR), leading to accumulation of Double‐Strand Breaks (DSBs). These observations led us to investigate the effects of long‐term exposure of intestinal epithelial cancer cells HCT116 to PMNs and PMN‐MPs. Consistent with the acute‐injury model, long‐term PMN‐MP treatment led to HR suppression with concomitant accumulation of DSBs, increased aneuploidy and apoptosis within 7 population doublings (PD7). With continued HR inhibition by PMN‐MPs, by PD12 we observed an upregulation of Non‐Homologous End‐Joining (NHEJ) activity. Importantly, DSBs and cell apoptosis were also significantly decreased at PD12, despite elevated aneuploidy, suggesting that PMN‐induced NHEJ activation promotes tumor cell survival and aneuploidy.

To test this in vivo, we employed a tumor xenograft model in conjunction with an antibody‐based PMN depletion strategy. Consistent with in vitro observations, HR inhibition, NHEJ upregulation, decreased apoptosis, and increased aneuploidy were confirmed in PMN‐infiltrated (PMN+) tumors as compared to PMN‐depleted (PMN−) tumors. To examine whether PMNs indeed provide survival advantage to tumor cells and help them overcome DSB accumulation, we performed ex vivo clonogenicity assay, where cells were challenged with DSB‐inducing agents Camptothecin (CMPT). PMN+ tumor cells were resistant to CMPT treatment and were able to more efficiently re‐propagate compared to parental HCT116 cells and PMN‐ tumor cells, which underwent apoptosis in response to CMPT‐induced genotoxic stress. Consistently, in PMN‐ tumor cells, upregulation of H2AX and cell cycle regulatory genes (p16INK4a, p27Kip1, TP53, p21Cip1) was observed, indicating inefficient DNA Damage repair. As such, our findings define a new regulatory role of PMNs in promoting CRC survival during carcinogenesis.

Support or Funding Information

This work was supported by grants from the National Institutes of Health (NIH) DK101675, Digestive Health Foundation, Chicago and by the Robert H. Lurie Comprehensive Cancer Center (Eisenberg Scholar grant) and by American Cancer Society to Ronen Sumagin.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Roles for Notch‐Sox9 in biliary epithelial cells‐to‐hepatocytes differentiation during liver regeneration
Authors: Ko Sungjin , Russell Jacquelyn O. , Shin Donghun Pal , Monga Satdarshan Pal
Published: June 2021

Liver regeneration after most forms of injury is mediated through proliferation of hepatocytes. However, when hepatocyte proliferation is impaired, such as during chronic liver disease, liver progenitor cells (LPCs) from the biliary epithelial cell (BEC) compartment can give rise to hepatocytes to mediate hepatic repair. Promotion of LPC‐to‐hepatocyte differentiation in patients with chronic liver disease could serve as a potential new therapeutic option, but first requires the identification of the molecular mechanisms driving this process. Notch signaling has been identified as an important signaling pathway promoting the BEC fate during development, and has also been implicated in regulating LPC differentiation during regeneration. SRY‐related HMG box transcription factor 9 (Sox9) is a direct target of Notch signaling in the liver, and Sox9 has also been shown to promote the BEC fate during development. We have recently shown that inhibition of Hdac1 activity during liver regeneration in both zebrafish and mouse models enhances sox9 expression in LPCs and impairs LPC‐to‐hepatocyte differentiation. Therefore, we hypothesized that inhibition of Notch signaling would promote LPC‐to‐hepatocyte differentiation by repressing sox9 expression. To this end, we blocked Notch activation during liver regeneration through treatment with γ‐secretase inhibitor LY411575 in a zebrafish LPC‐driven regeneration model and demonstrated enhanced induction of Hnf4a in LPCs. Alternatively, enhancing Notch signaling via Notch3 intracellular domain (N3ICD) overexpression impaired Hnf4a induction. Hepatocyte ablation in sox9b heterozygous mutant embryos enhanced Hnf4a induction, while BEC‐specific Sox9b overexpression impaired LPC‐to‐hepatocyte differentiation. Moreover, LY411575 treatment promoted the differentiation of murine LPCs into hepatocytes in vitro. Lastly, we administrated LY411575 to mice with hepatocyte‐specific deletion of β‐catenin, which induces differentiation of LPCs into hepatocytes following a choline‐deficient, ethionine‐supplemented (CDE) diet. We also examined the effect of liver‐specific Sox9 ablation upon CDE diet liver injury. Our results will elucidate the detailed roles for Notch‐Sox9 signaling in liver regeneration and LPC‐to‐hepatocyte differentiation.

Support or Funding Information

The work was supported by NIH grants to D.S. (DK101426), S.P.M. (DK62277, DK100287, CA204586), and J.O.R. (T32EB0010216, 1F31DK115017). S.P.M. is an Endowed Chair for Experimental Pathology.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Expression of NRG1‐ERBB4‐PSEN1‐NUP98 (NEPN) Signaling Axis as Potential Blood‐Based Biomarkers for Viral Myocarditis
Authors: Hanson Paul J , Lin Veena , McManus Bruce M
Published: June 2021

Background

Myocarditis, inflammation of the myocardium, is a leading cause of unexpected heart failure in young adults, commonly attributable to viral infections. Myocarditis manifests in a wide range of clinical presentations, from asymptomatic to acute heart failure. The current gold standard for diagnosis relies upon invasive endomyocardial biopsy, requiring histopathological demonstration of inflammation with/without associated myocyte damage. Under these guidelines, diagnostic sensitivity in independent published studies is <30%. We have previously demonstrated that detection of biomarkers within the NEPN pathway (implicated in immune response and heart failure) significantly improved diagnostic sensitivity to ~80%, however, this approach still relies upon biopsy. Thus, in the present study, our aim is to develop a non‐invasive, blood‐based diagnostic assay for viral myocarditis by examining the NEPN signaling axis and their cleavage fragments during the evolution of myocarditis.

Design

HeLa cells and human induced pluripotent stem cell (iPSC)‐derived cardiomyocytes (CM) were CVB3‐ or sham‐infected. Western blot analysis and confocal microscopy were performed to determine the expression and subcellular localization of NEPN signaling axis proteins in tissue culture. In addition, tissue and blood from murine models of viral myocarditis (A/J and C57BL/6 mice) were collected at 4 dpi (viremic phase) and 5 to 8 dpi (acute phase), and were analyzed by Western blot analysis, IHC and IF to detect protein expression.

Results

NEPN proteins all show changes in expression and subcellular localization throughout the infection time course. NUP98 is upregulated and cleaved during infection. NRG1 is upregulated in both lysates and supernatants. ERBB4 co‐localizes at the cell membrane with the protease PSEN1. ERBB4 and NRG1 cleavage fragments are secreted and detected in both tissue culture supernatant and mouse plasma. In the plasma of the less susceptible C57BL/6 mice, ERBB4 and NRG1 cleavage fragments increase throughout the viremic and acute phases. However, in the highly susceptible A/J mice, ERBB4 and NRG1 cleavage fragments were not detected in the plasma until 6 dpi (acute phase), when they are highly upregulated as compared to C57BL/6 mice (p=0.001).

Conclusion

Differential expression, subcellular localization and cleavage fragments of NEPN signaling axis proteins were detected during the pathogenesis of viral myocarditis. These composite changes may be useful in the development of a blood‐based diagnostic.

Support or Funding Information

Dr. Hanson's research is supported by the Myocarditis Foundation and Michael Smith Foundation for Health Research

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Understanding Leukocyte Recruitment in Murine Ozone‐Induced Lung Inflammation
Authors: Brocos Jessica Andrea , Aulakh Gurpreet Kaur , Snead Elisabeth , Singh Jaswant
Published: June 2021

Ozone, a highly reactive air pollutant, has been linked to a variety of acute and chronic respiratory diseases in humans. Health Canada guidelines suggest acute (1 h) ozone exposures should not exceed 120 ppb. Studies exposing mice to much higher levels of ozone (2 ppm) suggest significant recruitment of lung neutrophils and macrophages at 24 h. However, to our knowledge, no studies have been conducted at ambient ozone concentrations. Daytime “ground” ozone levels can easily reach 50 ppb. We report that ambient ozone, at 50 ppb, induces alveolar cell death, leukocyte recruitment and has the ability to cause lung damage and thus is of relevance to public health. We hypothesized that CX3CR1 macrophages are protective in murine ozone‐induced lung inflammation and mediate lung neutrophil recruitment. We exposed wild‐type and CX3CR1‐null mice to 50 ppb ozone or filtered air for 2 hours, and collected peripheral blood, lung vascular perfusate and bronchoalveolar lavage at 0 h, 6 h and 22 h after exposure. Our preliminary data suggest alveolar macrophage, epithelial and endothelial cell toxicity, alveolar and systemic nuclear chromatin and actin deposits, with vascular neutrophil recruitment being observed as early as 0 h, 6 h and 22 h after ozone exposure. Moreover, absence of CX3CR1 leads to an exaggerated inflammatory response marked by enhanced neutrophil recruitment. We aim to further characterize the cellular metabolic and cytokine responses in our model, which will allow us to better understand how and why leukocytes are recruited in ozone‐induced lung inflammation, and will guide development of protective health strategies and environmental standards.

Support or Funding Information

Fedoruk Centre and Innovation Saskatchewan

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.