bluuebunny/crossref_metadata_2025 huggingface dataset:
Preserving the extra cellular matrix is critical for improving efficacy of keratinocyte sheets for treatment of burn wounds
Authors: Alharbi Suzan , Niimi Yosuke , Popov Vsevolod , Hawkins Hal , Cox Robert , Wiener Dominique , Osada Atsuyoshi , Ihara Koji , Sakurai Hiroyuki , Herndon David , Enkhbaatar Perenlei
Published: June 2021

Introduction

Cultured autologous keratinocyte sheets (KS) have been proposed for treating burn wounds for over 30 years; however, they have not been translated to clinical practice because of negative effects of enzymes, used for detachment, on KS quality. The aim of this study was to test the hypothesis that non‐enzymatically detached KS exert better effects on grafted burn wound healing compared to enzymatically detached KS.

Methods

For non‐enzymatic detachment, autologous ovine primary keratinocytes were cultured in novel temperature‐responsive dishes (T sheets), and were detached by temperature reduction to 20°C. For comparison, the aliquots of keratinocytes were also cultured in conventional dish to be detached by use of enzyme (Dispase) (D sheets). We have tested the efficacy of these sheets using a well‐characterized, novel and clinically relevant ovine model of grafted burn wounds. Briefly, six full thickness skin burns (5×5cm) were induced in sheep dorsum under anesthesia and analgesia. After 24 hrs, burned skin was excised and wounds were grafted with ovine cadaver skin. After 3 weeks, rejected cadaver skin epidermis was debrided and wounds were covered with T and D sheets. Thereafter, the wounds were assessed for two more weeks. Wound samples were collected postmortem for various assays. The autologous keratinocytes for the sheet culture were isolated at the same day of skin grafting which mimics the clinical situation.

Results

Cultured keratinocytes in both dishes formed firm sheets within three weeks, mimicking the time of cadaver skin epidermis rejection. Keratinocyte sheet thickness was significantly higher in T sheets compared to D sheets. In vivo, T sheets were better accepted with significantly higher wound re‐epithelialization percentage than D sheets at days 7 and 14 after sheet grafting. Although the epidermis thickness was comparable in wounds treated with both types of sheets, the dermal‐epidermal junction was well defined in wounds covered with T sheets at 14th day as evidenced by continuous lamina densa and high number of hemi‐desmosomes. Histopathological evaluation for burn wounds revealed that wounds treated with T sheets had less hemorrhage, ulceration incidence and neutrophils infiltration.

Conclusion

Novel non‐enzymatic method for KS detachment provides better sheet quality and accelerates healing of grafted burn wounds.

Support or Funding Information

Shriner's Hospital for Children SHC85100 and SHC84050

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gut bacteria contributes to NAFLD pathogenesis by promoting secondary bile acids biosynthesis
Authors: Jiao Na , Wu Dingfeng , Yang Zihuan , Fang Sa , Li Xiaoyi , Yuan Meifei , Zhu Ruixin , Zhu Lixin
Published: June 2021

Objective

The gut microbiota affect liver metabolism and health through its metabolites. We previously observed that the microbiome may have a negative effect on non‐alcoholic fatty liver disease (NAFLD) through elevated secondary bile acids (BAs) production, and consequently suppressed BA‐mediated signaling. In this study, we examined the whole genome sequencing (WGS) data from NAFLD microbiome aiming to identify the species that contribute to the altered BA metabolism in NAFLD.

Methods

The metagenome data of the gut microbiota from 79 NAFLD patients were compared to those from 18 healthy controls. Metaphlan2 and HUMAnN were used to examine the gene‐based taxonomic and functional profiles of gut microbiota. Genome‐based analysis was further performed to obtain insights on the microbial genomes and to identify BA metabolizing microbes. Statistical tests, feature selection and microbial interaction analysis were integrated to identify disease markers and structural alterations in NAFLD gut microbiome.

Results

The gut microbiota of NAFLD were less diverse than those of healthy subjects. In addition, interactions among the members of the gut community in NAFLD were less than those in healthy subjects. We identified 37 differential species markers, including Ruminococcus torques, Clostridium leptum, and Escherichia coli; and 32 differential pathway markers including secondary BA biosynthesis and arachidonic acid metabolism, that distinguish NAFLD from healthy controls. Subsequently, 115 high‐quality microbial genomes (HQMGs) were re‐assembled, and taxonomy determined at strain level. Notably, metabolic potentials analysis identified 15 HQMGs, from Clostridium saccharolyticum and Ruminococcus bromii, that are involved in secondary BA biosynthesis, and were dominant in NAFLD and interplayed as a synergetic ecological guild.

Conclusions

In‐depth analysis of the WGS data from NAFLD patients and healthy controls revealed the structural and functional alterations of the gut microbiota. We identified the bacterial strains responsible for elevated secondary BA production in NAFLD, supporting our hypothesis that elevated secondary BA biosynthesis contributes to the development of NAFLD. The 15 BA‐metabolizing strains dominant in NAFLD could be novel therapeutic targets for NAFLD. In addition, our WGS study revealed strain‐specific functional differences within certain gut microbial species.

Support or Funding Information

This work was supported by National Natural Science Foundation of China 81770571 (to LZ), 81774152(to RZ), 41530105 (to RZ).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Autophagy Deficiency in the Liver Altered Pathogenesis of Alcoholic Liver Disease and Profile of Gut Microbiota
Authors: Yan Shengmin , Khambu Bilon , Chen Xiaoyun , Yin Xiao‐Ming
Published: June 2021

Autophagy is an evolutionarily conserved cellular degradation process that is critical to maintain metabolism homeostasis in the liver. To investigate the role of autophagy in liver diseases, we examined the impact of autophagy deficiency on ethanol‐induced liver injury and the gut‐liver relationship. First, hepatic autophagy deficient mice were challenged with ethanol by either acute or acute‐on‐chronic scheme. Our results demonstrated a protective role of autophagy in ethanol‐induced liver injury in a genetic and an inducible hepatic Atg7 knockout mice. Surprisingly, hepatic Atg5 knockout mice, which had a minor presentation of liver injury, were resistant to acute ethanol treatment, and were actually improved in the liver presentation following the treatment of in the chronic‐plus‐binge regime. A series of ethanol metabolism‐related genes were altered in livers of ethanol‐fed Atg5Δhep mice, which was accompanied with a faster clearance of plasma ethanol. This may partially explain the seemingly paradoxical observations. Second, gut microbiota was analyzed in fecal samples from hepatic autophagy deficient mice by 16S sequencing. The profile of gut microbiota was altered by hepatic autophagy deficiency with a notable increase of bile acids metabolism related bacteria. In addition to this, significant increase of total bile acids was observed in serum, liver, and feces. Moreover, ileal FXR was activated as increased gene expression of its target genes, including Shp, Fgf15 and Ibabp. Finally, modulation of gut microbiota by antibiotics treatment suppressed ileal FXR activation, increased enterohepatic bile acids pool, and aggravated liver injury in hepatic autophagy‐deficient mice. Collectively, our results indicate that hepatic autophagy has significant impacts on liver homeostasis and gut microbiota. The alteration of gut microbiota in hepatic autophagy deficiency can have in turn notable effects on liver injury.

Support or Funding Information

This research was partially supported by NIH R01AA021751 (X.‐M.Y.).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanisms of Gastrulation: from Zebrafish to Human
Authors: Solnica‐Krezel Lila
Published: June 2021

During animal gastrulation germ layers are induced, patterned and shaped into a body plan. These inductive and morphogenetic processes must be coordinated in space and time to ensure normal development, but the underlying mechanisms are not understood. I will discuss our forward and reverse genetic approaches in zebrafish that address this problem. I will also discuss our work with embryonic stem cells to investigate aspects of human gastrulation.

Support or Funding Information

1R35GM118179‐01 NIH/NIGMS Inductive and Morphogenetic Processes Shaping the Zebrafish Embryonic Axes

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Triggering Receptor Expressed on Myeloid Cells 2 (TREM2) Mutations: a Potential Common Cause of Alzheimer's Disease and Musculoskeletal Disorders
Authors: Plotkin Lilian I.
Published: June 2021

Alzheimer's disease (AD), a worldwide growing health issue, is a purely neurodegenerative form of age‐induced dementia. Interestingly, weight loss is frequently associated with disease severity and clinical progression and has been shown to begin at least ten years before the diagnosis of the disease. Bone frailty and sarcopenia have each also been linked to the onset of this disease, although very little is known about the pathogenesis of these conditions in subjects with AD. Indeed, although a common link between dementia and osteoporosis has been suggested, it has never been conclusively demonstrated. The prevalence of both conditions increase with aging, and some genetic conditions can lead to both AD and osteoporosis. Examples for the potential common cause of the 2 conditions are mutations of the Triggering Receptor Expressed on Myeloid cells 2 (TREM2) gene, primarily expressed in myeloid cells or microglia in brain and in osteoclasts in bone, which result in AD and dementia, as well as in bone fragility. In a recent Genome Wide Association Study, a single nucleotide polymorphism (SNP) encoding the R47H variant in TREM2 was linked with strong single allele genetic risk factors for AD, as well as with increased risk for Parkinson's disease, amyotrophic lateral sclerosis and frontotemporal dementia. In addition, recent studies showed that mice expressing the R47H TREM2 variant exhibit loss of TREM2 function and neuritic dystrophy. In a similar manner, another TREM2 mutation, the Y38C variant, was previously described in the pathogenesis of Nasu‐Hakola disease (also known as PLOSL) which is characterized by extensive white matter loss and frontotemporal‐like dementia and polycystic lipomembranous osteodysplasia. To investigate whether mutations in the TREM2 gene, previously associated with AD and frontotemporal dementia, also affect the skeleton, we analyzed the musculoskeletal phenotype of mice in which the Y38C and R47H variants of the TREM2 gene are ubiquitously expressed, generated by Drs. Lamb and Landreth at the Stark Neuroscience Research Institute, Indiana University School of Medicine. Our studies showed decreased bone mass in mice expressing the TREM2 mutants at 6–8 months of age. Interestingly, even though TREM2 expression was absent in skeletal muscle of wild type mice, mice expressing the R47H TREM2 variant exhibit decrease muscle mass. In summary, our results suggest a common cause of cognitive disorders and bone loss, and support an indirect role of TREM2 mutations on the loss of muscle mass, secondary to the altered bone remodeling.

Support or Funding Information

NIH/NIAMS R01AR067210

ICMH Collaborative Pilot Project Award

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Irisin Mediates Effects on Bone via αV Integrin Receptors
Authors: Kim Hyeonwoo , Wrann Christiane D. , Jedrychowski Mark , Kitase Yukiko , Nagano Kenichi , Strutzenberg Timothy S. , Bouxsein Mary L. , Griffin Patrick R. , Baron Roland , Rosen Clifford J. , Bonewald Lynda F. , Spiegelman Bruce M.
Published: June 2021

Exercise is crucial to prevent obesity, diabetes, fatty liver disease, and osteoporosis. These beneficial effects are partially mediated by myokines, hormone‐like molecules produced by skeletal muscle. Irisin is a myokine which is a cleaved product from fibronectin type III domain‐containing protein 5 (FNDC5) and is shed into the intracellular milieu and blood circulation to reach other organs. Particularly irisin has been shown to have beneficial effects in adipose tissues, brain, and bone. However, the skeletal response to exercise is less clear and the receptor for irisin has not been identified. Here we show that irisin binds to proteins of the αV class of integrins and biophysical studies identify interacting surfaces between irisin and αV/β5 integrin. Irisin increases both osteocytic survival and production of sclerostin, a local modulator of bone remodeling. Chemical inhibition of the αV integrins blocks signaling and function by irisin in osteocytes and fat cells. In vivo, injection of recombinant irisin protein increases the expression of sclerostin in osteocyte‐enriched bone as well as the levels of plasma sclerostin, and chemical inhibition of the αV integrins blocks the irisin‐induced effects. Genetic ablation of FNDC5/irisin completely blocks osteocytic osteolysis and bone resorption induced by ovariectomy, preventing bone loss and supporting an important role for irisin in skeletal remodeling. The identification of the irisin receptor and its roles in bone biology should greatly facilitate our understanding of irisin's function in exercise and human health.

Support or Funding Information

Hyeonwoo (Chad) Kim was supported by a cardiovascular T32 grant (5T32HL007374‐38). Bruce M. Spiegelman was supported by NIH grants (4 R01DK54477‐19, DK61502, B.M.S.) and the JPB Foundation (629803). Lynda F. Bonewald was supported by NIH grant (PO1AG039335). Clifford J. Rosen was supported by NIH grant (NIDDK RC2 092759).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Scoping Review: The Use of Augmented Reality in Medical and Surgical Anatomical Education and Its Assessment Tools
Authors: Lee Angela , McBain Kimberly , Ventura Nicole M , Noël Geoffroy PJC
Published: June 2021

Introduction

With the increasing accessibility to new technologies such as virtual reality and augmented reality (AR), a growing number of educators are exploring how to incorporate such advances to their field of study; anatomical education is no exception.

Aim

The purpose of this study was to identify the different AR modalities used to teach anatomy to students, medical/veterinarian trainees and surgeons via coursework, and/or procedural training. We also examined the qualitative and quantitative assessment tools used to evaluate the performance of various AR technologies in specific teaching settings.

Methods

A scoping review of the Web of Science, Pubmed, Embase and Medline was performed. Search terms were variations of 1) augmented reality, 2) medical or anatomical teaching/education/training, and 3) anatomy or radiology or cadaver. Abstracts, full articles, conference presentations, and guidelines published between January 2000–September 12, 2018 were identified and screened as per Preferred Reporting Items for Systematic Reviews and Meta‐Analyses (PRISMA) guidelines. Virtual reality was an exclusion criterion. The number of participants, level of training, name of AR modality, aim of the study, setting of the study (anatomy course vs. procedural training), object that was “augmented”, body system studied, type of assessment tools, and relevant findings were extracted from accepted studies.

Results

Preliminary findings suggests that Microsoft Hololens and projection‐based modalities using Microsoft Kinect were the most prevalent modalities. The studies were mainly conducted in the context of an anatomy course, which primarily assessed usability, learner satisfaction and perceived benefits of AR through questionnaires using variations of the Likert scale. Certain studies also incorporated more objective findings such as pre‐ and post‐AR knowledge tests. However, the majority of those studies failed to use validated tests.

Discussion/Conclusion

The current literature seems supportive of the use of AR as an adjunctive teaching tool at the very least. However, the evidence is weak given the lack of studies with robust quantitative and qualitative methodology to objectively determine the influence of the integration of AR on anatomical education. Sufficiently powered studies using validated assessment tools must be conducted to better understand the role of AR in anatomical education.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structure of auditory nerve synapses in normal and in hearing impaired
Authors: Rubio Maria Eualalia
Published: June 2021

The neurons of the cochlear nuclei in the brainstem are the first central processors of auditory information, and they provide inputs to all the major brainstem and midbrain auditory nuclei. The synaptic connectivity pattern of neural networks in each part of the cochlear nuclear complex is an essential determinant of their role in information processing. The synapse of the auditory nerve on principal neurons in the cochlear nucleus is biophysically and anatomically specialized for the fast conduction of excitatory currents into the auditory pathway. A growing interest in the field is to determine how auditory nerve synapses adapt to fluctuations in the sound environment. In this paper, we will discuss novel findings related to the structural and molecular responses of auditory nerve synapses and enwrapping astrocytes in the normal hearing and in the hearing impaired.

Support or Funding Information

NIDCD/NIH: R01DC013048; R56DC013048

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular mechanisms regulating presynaptic neurotransmitter release and their impact on neuronal circuit function
Authors: Young Samuel M
Published: June 2021

Deciphering the molecular mechanisms that govern neurotransmitter release at the presynaptic terminal is essential for understanding synaptic strength, plasticity, modulation and pathology. Voltage‐gated Ca2+ channels (VGCCs) regulate the magnitude and location of Ca2+ entry into presynaptic terminals of the central nervous system (CNS) and this determines the probability and kinetics of release of neurotransmitter‐filled synaptic vesicles (SVs) which ultimately impact neuronal circuit output and behavior. Located within the presynaptic terminal is the active zone, a dense network of proteins, which are implicated to regulate SV release by controlling VGCC function. However, the molecular mechanisms by which active zone proteins control VGCC density and organization within the presynaptic terminal are unclear. The calyx of Held, a glutamatergic presynaptic terminal in the auditory brainstem utilizes rapid and temporally precise action potential signaling for encoding information. Since it is the sole input to drive post‐synaptic action potential spiking and due to the ability to directly measure presynaptic Ca2+ currents and correlate them to SV release rates, the calyx is an exceptional model for gaining mechanistic insights into the presynaptic regulation of SV release and neuronal circuit output. We are using transgenic mouse models and novel viral vectors to manipulate presynaptic active zone proteins and CaV2 subtypes at the calyx during different developmental stages. In combination with these molecular manipulations, we utilized morphological analysis of VGGC organization in the presynaptic active zone and presynaptic ultrastructure and electrophysiology to determine their impact on synaptic biophysics, transmission and plasticity. Our results have identified key molecules and uncovered molecular mechanisms regulating CaV2 subtype levels, organization and proximity to SVs thereby controlling synaptic transmission and neuronal circuit output.

Support or Funding Information

Current Research Support: National Institute of Deafness and Communication Disorders (R01 DC014093), the University of Iowa.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Epigenetics and chronic pain
Authors: Stone Laura S
Published: June 2021

The emerging field of pain epigenetics will be introduced and the potential implications of this new field will be reviewed. Long‐term programming of gene expression is dynamically regulated by chemical modifications to the DNA and histones collectively referred to as epigenetic modifications. Epigenetic changes are responsive to the environment and have long‐lasting biological consequences. For example, early life neglect results in altered DNA methylation in the brain that are associated with maladaptive behavioural patterns in the adult. Evidence of epigenetic dysregulation in pre‐clinical pain models and in individuals with low back pain will be highlighted. Finally, the potential for epigenetics to understand biological mechanisms, to unmask new therapeutic targets and as biomarkers for chronic pain will be discussed.

Support or Funding Information

Supported by The Canadian Institutes for Health Research, The Quebec Pain Research Network and the Louise and Alan Edwards Foundation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structural, Functional and Gene Expression Analyses of the Aorta and Carotid Arteries in Newborn Term and Moderately Preterm Lambs
Authors: Black Mary Jane , Lombardo Paul , Nguyen Vivian , Flores Tracey , Sutherland Megan , Nitsos Ilias , Allison Beth , Parkington Helena , Tare Marianne , Harding Richard , De Matteo Robert , Schneider Michal , Polglase Graeme
Published: June 2021

Background

Preterm birth (< 37 weeks of gestation) occurs in approximately 10% of births worldwide; the majority of preterm births are moderately preterm (between 32 and 36 weeks gestation). We hypothesized that when birth occurs before term the major conduit arteries would be structurally and functionally immature and thus ill‐prepared for the hemodynamic transition that necessarily occurs at birth. Utilising a preclinical sheep model, our aims were to compare the structure and function of the thoracic aorta and common carotid arteries of moderately preterm (0.9 of gestation) and term‐born lambs in the immediate period after birth, and to determine whether there was upregulation of vascular injury‐associated genes following moderate preterm birth.

Methods

Time‐mated ewes were induced to vaginally deliver lambs either preterm (132 ± 1 days of gestation; n=11 females and n=10 males) or at term (147 ± 1 days of gestation; n=10 females and n=5 males). At 2 days after birth, ultrasound imaging of the proximal ascending aorta, main, right and left pulmonary arteries and right and left common carotid arteries was conducted in anaesthetised lambs. The lambs were then euthanized and segments of the thoracic aorta and left common carotid artery were either snap frozen for later real‐time PCR analyses or immersion‐fixed in 10% buffered formalin for histological quantification of collagen, smooth muscle and elastin within the medial layer.

Results

Overall there were few differences in vascular structure, wall composition and blood flow measurements of large conduit arteries at postnatal day 2 between moderately preterm and term lambs. However, there was a significant decrease in the proportion of collagen within the thoracic aortic wall (predominantly in males; p=0.048), narrowing of the carotid arteries (p=0.004) and a reduction in peak aortic systolic blood flow (p=0.049); all of which have the potential to impact on cardiovascular function if they persist long‐term. In addition, there was significantly increased mRNA expression of the cell adhesion marker P‐selectin in the thoracic aortic wall (p=0.001) and the pro‐inflammatory marker IL‐1β in the left carotid artery (p=0.04) in preterm lambs, suggestive of postnatal vascular injury.

Conclusions

The findings suggest that offspring born moderately preterm may be at increased risk of cardiovascular impairment later in life.

Support or Funding Information

This study was funded by the National Health and Medical Research Council of Australia

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glycobiology of Host‐Microbe Interactions in the Gut
Authors: Juge Nathalie
Published: June 2021

The gastrointestinal (GI) tract is colonised by trillions of bacteria collectively known as the ‘gut microbiota’ which have a profound impact on human health. Bacterial adhesion is a critical step for colonization of the host. Cell surface proteins (adhesins or lectins) mediate attachment to the host and initiate colonization by interacting with host proteins or glycoconjugates. However, in contrast to pathogens, the molecular basis for adhesion of commensal bacteria or probiotics to the gut remain poorly defined.

The mucus layer covering the GI tract is the first point contact between the gut microbiota and the host. There is an emerging paradigm that mucin glycans are key molecular determinants of gut homeostasis by providing binding sites and/or nutrients to the bacteria which have adapted to the mucosal environment.

Here we provide structural and functional insights into how gut bacteria adapt to the GI tract by focusing on the adhesion strategies of two inhabitant species of the GI tract, Ruminococcus gnavus1,2 and Lactobacillus reuter,3,4,5. The presentation will discuss relevant mechanistic insights into the nature, structure and function of bacterial adhesins involved in the interaction between these gut symbionts and the host glycoconjugates. Our results showed that adhesion strategy is species‐ and strain‐specific and occurs via specialised mucus‐binding proteins or carbohydrate‐binding modules. Sialic acid, a terminal epitope in intestinal mucins, plays a critical role in mediating bacterial tropism along the GI tract.

Exploring the molecular basis of adhesion by resident species of the gut microbiota is important for the design of microbiota‐targeted strategies to restore gut health.

Support or Funding Information

The work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC), under the Institute Strategic Programme Grant for Gut Microbes and Health (BB/R012490/1).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of Ancestry Genes in Asthma Susceptibility
Authors: Ortega Victor
Published: June 2021

Adverse racial differences exist throughout respiratory diseases, including asthma where African descent minority groups such as African Americans and Puerto Ricans experience greater asthma risk and morbidity compared to European descent Whites. Advances in genomic technologies have advanced the complexity and quantity of genetic data to analyze and interpret in individuals from different racial groups with varying inter‐individual ancestral backgrounds. The vast majority of genomic studies have been performed in European descent Whites, but there has been increasingly diverse studies consisting of different racial and ethnic groups where differences have been demonstrated in asthma risk, severity, and drug response. A “bottleneck” resulted in a collapse of genetic diversity following the first human colonization of Europe during the Upper Paleolithic period and the more recent mixing of African, European, and Native American ancestries. These recent admixture events have resulted in different ethnic groups with varying degrees of genetic diversity, including variability in genetic tagging architectures and the frequency of genetic variants, particularly rare variants which are most frequently found to be unique to individual ancestries. Variability in genetic ancestral background could be associated with variation that has the potential to alter asthma risk, severity, or therapeutic responsiveness to commonly used therapies. Genomic studies have employed both traditional and admixture‐based whole‐genome approaches in minority, ethnically diverse cohorts resulting in the confirmation of genomic discoveries initially only found in Whites while identifying novel genetic determinants of asthma risk and drug response unique to specific ethnic groups. Large consortium‐based sequencing studies utilize next‐generation whole‐genome sequencing in diverse multi‐ethnic cohorts to provide a map of the genomes of different recently admixed populations to guide future genomic studies. Genomic studies in increasingly diverse cohorts provide an opportunity to facilitate discoveries and are a critical step towards precision medicine applicable to all people.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Opportunities and Challenges for Inter‐Professional One Health Education
Authors: Castellot John , Afifi Linda
Published: June 2021

One Health is an interdisciplinary approach to optimizing health for humans, animals, and the environment at the local, national, and global level. It incorporates expertise from diverse fields—including human and veterinary medicine, environmental and biological sciences, engineering, public health, political science, urban planning, economics, sociology, epidemiology, and statistical modeling—to study and solve important challenges at the intersection of humans, animals, and the environment.

While it is quite easy to get buy‐in from the various stakeholders in One Health, there are many barriers to incorporating inter‐professional education in this area. For example, many undergraduate institutions do not have public health, health profession, and engineering schools. Even universities like Tufts, which has these schools, face significant obstacles in integrating One Health into their curricula.

Specific suggestions for overcoming these obstacles will be provided, including seed grant initiatives and mini‐symposia, along with a description of a highly successful inter‐professional One Health activity currently ongoing at our institution: a student‐driven, faculty coordinated “Comparative Anatomy Exchange Day”. This activity involves veterinary, medical, dental, physician assistant, and graduate students.

The event is hosted at the medical school, which has a newly‐built Gross Anatomy laboratory. Since human anatomy labs have tables sized for humans, it is not possible to accommodate large animal cadavers. Thus, the focus is on a specific organ system such as the GI system. The vet students bring GI systems from large animals, including horses, cows, camelids, etc; smaller animal cadavers such as birds, cats, dogs, pigs, and sheep are also provided. The medical school provides 1–2 previously dissected cadavers that are especially good for illustrating the GI system.

The morning session is devoted to students from the various professional schools presenting the GI system of the human and animal species via a series of short, pithy talks. Following lunch, all of the participants head to the Gross Anatomy laboratory where the human and animal cadavers are displayed. Students go from station to station as they wish. A student is in charge of describing the relevant anatomy at each station, with a small number of faculty available for questions.

This activity has proven to be highly popular, despite the fact that it is an optional activity offered on a Saturday afternoon in late April—more than 130 students and faculty participated. Creating One Health activities that are fun, educational, and allow students from different professions to interact is a highly effective way to provide inter‐professional education around One Health.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Signals, forces, and cells: Decoding tissue morphogenesis
Authors: Zallen Jennifer
Published: June 2021

A major challenge in developmental biology is to understand how changes in tissue structure are generated by processes that occur on a cellular and molecular scale. In Drosophila, the characteristic elongated shape of the body axis is achieved through the coordinated movements of hundreds of cells. We discovered that these movements are oriented by subcellular asymmetries in the localization and activity of proteins that generate contractile and adhesive forces within cells. In addition, we identified a positional code that systematically orients cell movements throughout the embryo and demonstrate that this spatial information is provided by an ancient family of Toll‐related receptors that are widely used for pathogen recognition by the innate immune system. Using quantitative live imaging and automated cell tracking, we showed that these molecular asymmetries result in dynamic, collective cell behaviors in which multicellular rosette structures form and resolve directionally, driving tissue elongation. Planar polarized actomyosin contractility and rosette behaviors have now been shown to promote tissue elongation in chicks, frogs, and mice, and represent a general mechanism linking cellular asymmetry to global tissue reorganization. We are currently using genetic, cell biological, biophysical, and live imaging approaches to understand how genes encode the forces that shape tissues, and to elucidate the mechanotransduction mechanisms that allow cells to modulate their behavior in response to changes in their mechanical environment.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Integration and the genetics of variation in facial shape
Authors: Hallgrimsson Benedikt , Aponte Jose David , Katz David C , Bannister Jordan J , Green Rebecca M , Young Nathan M , Cheverud James M , Forkert Nils Daniel , Marcucio Ralph S
Published: June 2021

Human facial form is both highly variable and heritable. Despite considerable effort, however, the genetic basis for the strong and often very particular patterns of resemblance among relatives are largely unknown. The apparently highly polygenic basis for facial shape variation appears to be at odds with the strong and often highly specific patterns of heritability for facial features. Facial shape variation tends to be structured around developmentally determined patterns of correlated variation. This tendency for covariation or integration results in morphological variation that occupies only a fraction of it possible dimensions. Synthesizing results from experimental and quantitative genetics of the face and applying multivariate genotype‐phenotype mapping, we argue that the integrated nature of variation in facial form can simplify qualitative explanations of the genetics of face but greatly complicate the ability to quantitatively relate genetic to phenotypic variation. Acting on variation in development, many genetic variants push or pull variation along the same limited number of dimensions that correspond to patterns of covariation among aspects of craniofacial variation, producing complex and overlapping patterns of pleiotropic effects. Developmental nonlinearities and interactions among processes at multiple levels result in gene interaction effects that can result in specific genetic variants having genetic background specific effects whilst having small and difficult to detect effects on variation at the population level. These findings have important general implications for the genetics of complex traits.

Support or Funding Information

CIHR Foundation Grant, NSERC #238992‐17, NIH R01DE019638; U01DE024440

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FAM20C is Essential for Maintaining Brain Homeostasis
Authors: Zhang Hua , Lu Yongbo , Qin Chunlin
Published: June 2021

OBJECTIVES

FAM20C (Family with sequence similarity 20, member C) mutations cause Raine syndrome in humans, characterized by generalized osteosclerosis, distinctive craniofacial dysmorphism, along with extensive intracranial calcification. While much progress has been made in understanding the role of FAM20C in osteogenesis and odontogenesis, the function of FAM20C in brain is still unclear. The goal of this study was to determine whether FAM20C is required for normal brain homeostasis in vivo.

METHODS

1) Reverse transcription polymerase chain reaction (RT‐PCR), Western‐blotting and in situ hybridization analyses were used to examine expression patterns of FAM20C in the mouse brain. 2) Global and conditional FAM20C‐knockout mice were generated by crossing FAM20C loxP mice with sox2‐cre (global) mice, Nestin‐cre (active in neuronal and glial cell precursors in brain) mice and 2.3 Col I‐Cre (active in osteoblasts/osteocytes in bone) mice, respectively. 3) A combination of histology, μ‐CT, and x‐ray methods were used to characterize the brain and bone phenotypes of the global and conditional FAM20C‐knockout mice.

RESULTS

1) FAM20C was broadly expressed in the neuronal cells in the brain. 2) The global deletion (mediated by Sox‐2 cre) and brain‐specific (mediated by Nestin‐cre) deletion of Fam20C resulted in cerebral calcification in mice after postnatal 3 months and 6 months, respectively. Interestingly, although the bone‐specific (mediated by 2.3 Col I‐Cre) deletion, like the global deletion, of Fam20C caused similar hypophosphatemic rickets/osteomalacia in mice, it did not lead to intracranial calcification after postnatal 6 months. 3) X‐ray and histological analyses showed that the calcifications were bilaterally distributed within the brain. These calcifications were not associated with dystrophic changes. There were mild perifocal microgliosis as well as astrogliosis around calcospherites.

CONCLUSIONS

The local loss of FAM20C function in the brain may directly account for intracranial calcification. FAM20C may play an essential role in maintaining normal brain homeostasis and preventing ectopic brain calcification.

Support or Funding Information

NIH/NIDCR DE022549 to CQ

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 4th Dimension of Transcriptional Networks: TIME
Authors: Coruzzi Gloria , Varala Kranthi , Marshall‐Colon Amy , Brooks Matthew , Ruffel Sandrine , Alvarez Jose , Pasquino Angelo , Cirrone Jacopo , Shasha Dennis
Published: June 2021

Our study exploits time ‐ the relatively unexplored dimension of gene regulatory networks (GRNs) ‐ to learn the temporal transcriptional logic that underlies dynamic nitrogen (N) signaling in plants [1, 2]. Because causality moves forward in time, fine‐scale time‐series experiments are a valuable resource to infer GRN models that can predict TF‐target relationships in out‐of‐sample data ‐ the ultimate goal in systems biology [3]. First, using fine‐scale time‐series transcriptome datasets from N‐treated plants we developed a in a “just‐in‐time” analysis approach that uncovered a temporal cascade of cis‐regulatory elements underlying dynamic N‐signaling [2]. Next, to learn the associated transcription factor (TF) cascade, we inferred a GRN using a time‐based machine learning method ‐ Dynamic Factor Graph ‐ capable of predicting future gene responses under untested conditions [1]. We performed network validation studies using TF perturbations in plant cells using the TARGET system [4], and also in planta [2]. Our TF‐target validations of seven TF hubs in this predicted network were used in a precision/recall AUPR analysis to identify high‐confidence TF‐target edges in the time‐inferred GRN [1]. This high‐confidence GRN recovered TFs previously validated in the N‐response, as well as novel ones that were validated to regulate 15NO3−‐uptake and assimilation in planta [1]. This validated time‐based GRN now provides the temporal transcriptional logic and high‐confidence target predictions for >150 TFs in the dynamic N‐signaling response in Arabidopsis [1, 2]. More broadly, this time‐based approach ‐ can be applied to uncover the temporal transcriptional logic for any signaling response system in biology, agriculture or 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.
Genomic and Proteomic Signatures of Replication Fork Collapse
Authors: Shastri Nishita , Rivard Rebecca S. , Yeng Ya‐Chu , Anastassiadis Theonie , Kulej Katarzyna , Jagannathan Veena , Mandal Rahul , Borole Piyush , Garcia Benjamin , Belinsky Anja‐Katrin , Brown Eric
Published: June 2021

Transient obstruction of DNA polymerase progression activates the ATR checkpoint kinase, which in turn suppresses fork breakage and DNA repair‐factor recruitment. To characterize the causes of and responses to replication fork collapse driven by inhibition of ATR, two unbiased, information‐rich approaches were applied: genome‐wide identification of replication fork collapse sites and quantification of protein recruitment to such sites. Over 500 murine and human sites in the genome were identified as dependent on ATR for stability. The vast majority of these sites were characterized by repetitive DNA, including quasipalindromic AT‐rich repeats, inverted retrotransposable elements and purine‐rich ministatelites. While the prevalence of these types of break‐associated repeats in human and mouse cell lines differed, the formation of stable intrastrand structures was a common feature. These data indicate that intrastrand structure formation is a primary driver of fork collapse upon ATR inhibition. Using a novel two‐step replication fork retrieval approach (iPOND2), we have gone on to show that the response to replication fork collapse involves both the rapid recruitment of various repair factors and subsequent large‐scale ubiquitin‐driven turnover of these same factors. We speculate that the establishment of this dynamic equilibrium of repair factors may allow the best repair mechanism for the specific type of damage and intermediate generated to gain access and be favored. These findings and how distinct types of DNA repeats could drive differing repair responses will be discussed.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antibody Recognition of a Polysaccharide Common to many Microbes and Biofilms
Authors: Soliman Caroline , Walduck Anna K. , Yuriev Elizabeth , Richards Jack S. , Cywes‐Bentley Colette , Pier Gerald B. , Ramsland Paul A.
Published: June 2021

Hospital‐acquired infections and multidrug resistant microbes represent a healthcare crisis where alternative treatments such as therapeutic antibodies and vaccines are needed. Poly‐N‐acetyl‐D‐glucosamine (PNAG) is a polysaccharide that is produced by many microbes, including bacteria, fungi, and protozoan parasites. Antibodies that interact with PNAG have demonstrated efficacy against a wide range of microbes. A human monoclonal antibody (mAb) F598 that binds PNAG has shown opsonic and protective activity against multiple microbial pathogens and is undergoing clinical assessment as a broad‐spectrum antimicrobial therapy. We recently determined high resolution crystal structures of the unliganded F598 antigen binding fragment (Fab) and its complexes with N‐acetyl‐D‐glucosamine (GlcNAc) and a PNAG oligosaccharide [Soliman et al (2018) J Biol Chem 293:5079]. Recognition of PNAG occurs in a large groove‐shaped binding site that traverses the entire light and heavy chain interface and accommodates at least 5 GlcNAc residues. The Fab‐GlcNAc complex revealed a deep binding pocket with near identical positioning of the monosaccharide and a core GlcNAc of the oligosaccharide suggesting an anchored binding mechanism of PNAG by the F598 mAb. Although intact F598 IgG is required for effector functions, the Fab used here retained binding to PNAG on the surface of an antibiotic‐resistant biofilm‐forming strain of Staphylococcus aureus. A model of intact F598 IgG binding to two pentasaccharide epitopes shows the antibody can span across an extended PNAG chain. This study reveals the structural basis for antibody recognition of PNAG on surfaces of microbial pathogens and in biofilms.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ITGB1‐DT and ITGB1 enhance lung cancer cell metastatic abilities and are associated with poor prognoses in NSCLC patients
Authors: Lai Tsung‐Ching , Hsiao Michael
Published: June 2021

Non‐small cell lung cancer (NSCLC) is one of the major leaders in lethal cancer for decades. It has two major types: lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). In the molecular analysis, the genomic backgrounds were different among two sub‐types. But most deaths from these two types of NSCLC were due to the metastatic cells which present different molecular characteristics compared to the primary cells. The differences may provide a key for novel diagnosis and/or therapeutic strategy to treat NSCLC. Here in this study, we have found a long non‐coding RNA, ITGB1‐DT (RP11‐462L8.1) and its neighboring gene, ITGB1 were found associated with poor prognosis of overall survival and recurrence‐free survival analysis from TCGA lung cancer cohort in both LUAD and LUSC. We determined the expression levels with our own clinical cohort and confirmed the findings. We also found that the ITGB1‐DT RNA expression levels were correlated to the higher invasion and migration abilities in CL1‐5 lung cancer cells compared to lower invasion and migration ability parental CL1‐0 cells. It is also involved in epithelial‐mesenchymal‐transition in A549 after TGF‐beta treatment. ShRNA induced inhibition of the ITGB1‐DT gene expression was found to reduce the invasion and migration ability of the highly aggressive NSCLC cells with endogenous ITGB1‐DT overexpression. Knockdown of IGTB1‐D1 in lung cancer cells also significantly suppressed cancer cell invasions and metastases in in‐situ human lung cancer established NSG mouse model. The putative molecular mechanisms of ITGB1_DT and ITGB1 in promoting lung cancer cell metastasis were elucidated. Together, our results identified a new long non‐coding RNA ITGB1‐DT who may interact with its neighbor coding gene ITGB1 to promote lung cancer cell metastasis. Inhibition of ITGB1‐DT and ITGB1 expressions and interactions may provide a novel treatment strategy to combat lung cancer patients with metastasis.

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