bluuebunny/crossref_metadata_2025 huggingface dataset:
One Plate by GAUTIER D'AGOTY on the Anatomy of the Frogs, Engraved in 1752 in Paris by Delaguette
Authors: Le Floch‐Prigent Patrice Pierre , Nguyen Juliette , Gillot Jean‐Bernard
Published: June 2021

An engraving of the frogs with seven figures was anatomically observed. It was 14,5 cm wide and 20,5 cm high. It contained two dissected male and female specimens, the dissections of the genito‐urinary apparatus in both sexes, one entire, living frog and a tadpole. This plate came from a journal directed and published by Jacques‐Fabien GAUTIER D'AGOTY (1716–1785): “Observations sur l'histoire naturelle, sur la physique et sur la peinture” (observations of the natural history, the physics and the painting). It accompanied the Observation # XX “sur l'anatomie des grenouilles” (on the anatomy of the frogs). These plates were originally in the in‐4 size; later they were reproduced for some of them in the in‐12 size. GAUTIER D'AGOTY was very famous for a procedure of engraving in four colors and the publication of a large book with muscular plates and also for full size spectacular, obstetrical, anatomical women.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Workflow for Successful Collaboration between Anatomists and Pathologists: An Example of Pulmonary Adenocarcinoma Metastasis to a Virchow Node
Authors: Zdilla Matthew J , Aldawood Ali M , Plata Andrew , Vos Jeffrey A , Lambert H. Wayne
Published: June 2021

Interprofessional collaboration between anatomists and pathologists is important, yet poses unique workflow challenges. Therefore, this report details a successful workflow example of a case of pulmonary adenocarcinoma metastasizing to a Virchow node wherein anatomists and pathologists collaborated in a manner that was both streamlined and effective. Essentially, gross pathology was identified in a gross anatomy laboratory, and an anatomist forwarded a description of these findings to the director of pathology residents. Next, the director of pathology residents forwarded basic details of gross anatomical findings to pathology residents while asking for resident participation based upon professional interests. The interested residents then contacted the anatomist, who, in turn, walked the residents through the gross anatomical findings in the dissection facilities. The residents resected the pathological tissues and then provided the anatomist with a written histopathological assessment that was confirmed by a seasoned pathologist. The insights of both the anatomists and pathologists led to successful description of the case with a robustness that either party would not have been able to complete on their own. This collaborative project led to a publication containing improved descriptions of clinically important findings arising from gross anatomy laboratories, and the effectiveness of the described workflow between anatomists, pathology residents, and pathologists will increase the collaborative output of all involved parties in future research projects and variant anatomical discoveries.

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 Size and Shape of the Porus Trigeminus: Implications for Trigeminal Neuralgia Procedures
Authors: Nestor Nicholas , Ritz Brianna , Hunter Dawn , Zdilla Matthew
Published: June 2021

Trigeminal neuralgia is characterized by debilitating facial pain. The treatment for trigeminal neuralgia often involves the introduction of a surgical tool through the foramen ovale and porus trigeminus (PT). Though several studies have recently described the anatomy of the foramen ovale, few have described the PT. This study analyzed 53 PT for area, height, width, and circularity among both left‐ and right‐sided PT from females and males. There were no statistically significant differences in the area, height, width, and circularity of the PT between sexes (U=258, p=0.42; U=255, p=0.38; U=298, p=0.99; U=295.5, p=0.95). Similarly, paired t‐tests revealed no significant differences between sides with regard to area (t=1.35; p=0.19) and height (t=0.40; p=0.69). However, there were significant differences between sides with regard to width (t=2.19; p=0.04) and circularity (t=3.38; p=0.003). The average left‐sided PT width was 6.6±2.4mm (Mean±SD) whereas the average right‐sided PT width was 7.7±1.6mm. With regard to surgical implications, right‐sided PT may provide slightly greater freedom with regard to the angle of approach to the PT.

Support or Funding Information

Funding: WV Research Challenge Fund [HEPC.dsr.17.06]

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 Book of LE CAT, “Traité des Sens” (= Treatise of the senses), Published in 1740, in Rouen (France)
Authors: Le Floch‐Prigent Patrice Pierre
Published: June 2021

This book contained 16 anatomical and optical plates with 3 double ones: the sketch and explanations on one side and the print on the other one. The engravings were made by Herisset. It is the rare, first edition about the five senses, particularly the vision's one. The book was an in octavo and contained 301 pages. There was a dedicacion to the lords of the Normandy's parliament. In fact it was only one part of a large book in three volumes which will be edited later in 1767 as “Oeuvres physiologiques, Traité des sensations et des passions en général, et des sens en particulier” (= Physiological works, Treatise of the sensations and of the passions in general, and of the senses in particular (Paris, by Vallat‐La‐Chapelle). Claude‐Nicolas LE CAT (1700–1768) founded the Royal Academy of Rouen and was one of the most famous french surgeon in the 18th century. He establishes his activity in the town of Rouen (a large harbour in the entrance of the Seine river) in 1729 as the surgeon of his protector: the archibishop La Vergne de Tresan, then he became the chief surgeon of the Hotel‐Dieu in Rouen in 1731 where he built a dissection amphitheater in march 1736. This skilful lithotomist (ablation of the bladder stones) was a member of several societies (Academies of Paris, London, Madrid, …). We specially analyzed the anatomical accuracy of the plates.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Decoupling of biomechanics of hyoid movement and bolus flow in preterm infants
Authors: Gould Francois , Catchpole Emily , Bond Laura , Stricklen Bethany , Mayerl Christopher , German Rebecca
Published: June 2021

Determining the timing of bolus passage over the airway, and into the esophagus, is critical for biomechanical studies of swallowing. Videofluroscopy is the gold standard for determining fluid movement, but is not always possible. Hylolaryngeal movement, measured from light film or standard videography, is a reliable marker for adult human swallowing, but its validity for the biomechanics of swallowing in immature infants is unknown. We tested the relationship between hyolaryngeal movement from light camera data compared to bolus movement from video fluoroscopy in pre‐term and term infant pigs.

Three term and four pre‐term pigs were fed from a bottle containing barium and a milk solution. At seven days after birth pigs were recorded feeding from the bottle using simultaneous high speed (100 fps) x‐ray and light video cameras. In the light video, the frame of most rapid elevation of the hyoid was scored as the swallow. In the x‐ray, the swallows were scored as the beginning of posterior bolus movement. The number of frames separating x‐ray and light camera swallow identifications was calculated for each swallow.

Hyoid movement was found to be closely correlated to bolus movement in term pigs, but no correlation existed in pre‐term pigs. The mean frame offsets for term and pre‐term pigs were different (Wilcox test, p<0.001). The variance for the pre‐term pigs was significantly larger (Levene's test p<0.001).

The light videos were found to be a good measure of swallows in term pigs, but were not a good measure of swallows in pre‐term pigs. The biomechanics of bolus movement are altered in the neurologically poorly coordinated pre‐term pigs. Specifically, hyolaryngeal elevation and bolus movement are decoupled in pre‐terms whereas they are tightly coordinated in term pigs. Prematurity leads to altered form function relationships through immature development.

Support or Funding Information

NICHD grant 088561

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evolutionary Ontogeny of the Avian Adductor Musculature‐Endocranial Relationship: Evidence from the Chick
Authors: Llera Catherine J , Bever Gabriel S
Published: June 2021

The modern avian body evolved across a stem lineage that is more than 140‐million years in length and is famously populated by a multitude of fossil forms, including all non‐avian dinosaurs. Expansion of the brain and surrounding neurocranium is an important driver of many morphological patterns, including the general reduction and ventral displacement of the adductor chamber. Nevertheless, a recent study showed that Ichthyornis — a Cretaceous stem bird that retains teeth and lies phylogenetically just outside the crown radiation — expresses both a derived braincase and a primitive adductor chamber. This unexpected co‐occurrence suggests complexity in the evolutionary trade‐off between these functionally important cranial spaces. We build on this work by examining the relationship between the developing adductor chamber and the endocranial cavity; specifically testing for temporal shifts as the adductor chamber develops and the skull grows around the precocial brain. Our sample consists of differentially stained μCT scans from a chick (Gallus) ontogenetic series that ranges from the Day 5 embryo to adult. The adductor chamber musculature and associated bones were digitally segmented. Muscle attachments at each stage and the volume for each individual muscle/undifferentiated muscle group were recorded. Results show the adductor muscle groups (adductor mandibulae posterior, externus, and internus) were clearly distinguishable by Day 15 of embryonic development. The individual muscle bellies within each group remained largely undifferentiated until postnatal week 6. Nonetheless, only in the adult stage are the muscle bellies within each group fully discernible. Analyses of muscle locations relative to skull bones show that attachment points tend to shift ventrally during development. Additionally, preliminary results indicate that the increase in muscle mass standardized by head weight is non‐linear and the relative adductor muscle mass is smaller as the chick matures. These results indicate an ontogeny‐phylogeny parallel, with the adductor muscles decreasing in relative size and shifting ventrally as the braincase expands. The timing and tempo of adductor muscle differentiation and maturation, and the morphological dynamics of the other feeding‐related structures (e.g., lower jaw) has potential implications for understanding the evolutionary trade‐off that occurred between brain expansion and reduced chewing musculature. Our data will serve as baseline trajectories for future developmental studies, including modelling how heterochronic or heterotopic alterations explain observed‐ phylogenetic patterns and predict future fossil discoveries.

Support or Funding Information

NSF‐DEB‐1801224 to GSB

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Caloric Restriction Increases Parkin Recruitment in Hearts of Obese Insulin‐Resistant Rats
Authors: Cummings Chelsy Tiy , Cornejo Manuel A , Nguyen Julie , Cazaress Joshua , Escobedo Benny , Nishiyama Akira , Ortiz Rudy M
Published: June 2021

Mitochondria provide energy to the cells, making the task of mitochondrial recycling highly important. Impairments in mitochondrial number and function can lead to dysregulation of cellular metabolism and disrupted capacity to maintain homeostasis. The Parkin gene codes for the protein, Parkin, which is a component of the Parkin/PINK1 pathway that helps regulate mitophagy. An impairment in the Parkin gene may also results in nonfunctional mitochondria, potentially leading to many diseases such as type two diabetes mellitus and cardiovascular disease. Recent studies suggest that Parkin expression is downregulated during insulin resistance. To address the hypothesis that insulin resistance downregulates Parkin in the heart, a piece of the left ventricle of the hearts of 4 groups of rats were extracted: (1) Long Evans Tokushima Otsuka (LETO), (2) Otsuka Long‐Evans Tokushima Fatty (OLETF), (3) LETO with caloric restriction, and (4) OLETF with caloric restriction. Extracted Parkin from the hearts of each group was measured by Western blot. Because Parkin expression is downregulated with insulin resistance, we anticipated lower levels of express in the insulin resistant, OLETF rats compared to LETO, and that caloric restriction would restore Parkin expression in the OLETF rats. Parkin protein expression was 71% lower in OLETF than LETO. While the 16% increase in expression in caloric restricted LETO group was not statistically significant, the expression increased nearly 7.5‐fold with caloric restriction in OLETF group compared to the ad lib OLETF group. The data suggest that the benefits of CR on CV function and health go beyond the potential benefits of mass loss (usually adiposity) but include cellular energetics and metabolism in the heart of insulin resistant rats.

Support or Funding Information

The STRIDE Fellowship is supported by the American Physiological Society and a grant from the National Heart, Lung and Blood Institute (NHLBI; 1 R25 HL115473‐01).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessing Post‐Stroke Cardiac Dysfunction in the Insular Ischemic Stroke Rat Model
Authors: Jaremek Victoria , Iankov Stella , Milazzo Thomas , Wang Lynn , Whitehead Shawn N. , Sposato Luciano A.
Published: June 2021

Patients who have suffered an ischemic stroke with damage to the right insular cortex (IC) often develop post‐stroke cardiac complications. However, the pathophysiology and time course of these post‐stroke myocardial changes require further investigation which is complicated by a lack of pre‐clinical models. The purpose of this work is to assess the time course of post‐stroke cardiac fibrosis and inflammation in a rat model of selective insular ischemic stroke. IC ischemic stroke was induced in six‐month‐old Wistar rats via unilateral stereotaxic injection of endothelin‐1 into the right IC (n=6/surgical group). Control rats received a phosphate‐buffered saline (PBS) injection into the right IC (n=6/surgical group). To establish a time course of cardiac dysfunction, hearts were histologically examined at 6, 24 hours, 7, 14 and 28 days post‐stroke for fibrosis (Masson's Trichrome stain), inflammation (CD45+, myeloperoxidase, CD3+, CD45R, CD68+ immunostaining) and endothelial dysfunction (phosphorylated endothelial nitric oxide synthase (eNOS)). Areas of interest included the 4 heart chambers and pulmonary vein/left atrium border (PV‐LA border). Results from this study showed that focal IC ischemic stroke led to neutrophil infiltration at 6 hours, followed by T lymphocyte infiltration at 7 days at the PV‐LA border. Left atrial tissue experienced long‐term fibrosis at 28 days following focal IC stroke. Further, B lymphocyte infiltration was seen at 28 days post‐stroke in left ventricular tissue. These findings provide insight into the progression of post‐stroke cardiac changes and suggest that inflammation is a treatable target to prevent post‐stroke myocardial injury.

Support or Funding Information

This study is funded by the Kathleen & Dr Henry Barnett Research Chair in Stroke Research (Western University, London, Ontario, Canada), the Western University Medical and Health Sciences Research Board Seed Research Grant, and the Edward and Alma Saraydar Neurosciences Fund (London Health Sciences 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.
The Silent Treatment: miR199 Family Silences Shh during Craniofacial Development
Authors: Richbourg Heather Ashley , Hu Diane , Marcucio Ralph
Published: June 2021

Early craniofacial development requires precise timing and spatial regulation of key signaling pathways in order for proper facial structures to form. Perturbations, particularly in the sonic hedgehog (Shh) signaling pathway, can result in craniofacial birth defects that cause significant harm and financial costs. microRNAs (miRNA) play many roles in developing systems by repressing gene expression, but despite the enormous number of miRNAs that have been identified, few have been related to biological processes through direct targets. Here we identified the miR199 family as Shh‐regulated miRNAs during development of the upper jaw in a chick model.

We performed a miRNA microarray analysis on chick embryonic ectoderm before (stage HH18) and after (stage HH22) formation of the Frontonasal Ectodermal Zone (FEZ) and after activating or blocking SHH signaling in the brain. We identified a family of miRNAs (miR199b, mir199* and miR199) that appeared to be regulated by SHH signaling in the brain. These are increased between stages HH18 and HH22, and increase upon activation of Shh signaling and decrease upon blockade of Shh signaling, and these data were confirmed by qPCR. To assess the function of this family of miRNAs during facial development, we performed miR199 knock‐down and overexpression experiments and examined the phenotypic and molecular changes. Up‐regulation of each miRNA led to decreased Shh expression in the FEZ and facial malformations that are consistent with decreased Shh signaling in the face. Using antagomirs to knockdown each miRNA, we observed that Shh expression was expanded and the faces were wider, reminiscent of SHH gain‐of‐function experiments. Hence, it appears that expression of these miRNAs in the FEZ are regulated by SHH signaling from the brain, and these miRNAs appear to regulate SHH signaling from the FEZ. We then identified Smurf2, Map3k4, Hif1alpha, and Dyrk1a as targets of the miR‐199 family of miRNAs via mir‐base. We confirmed that each of these genes behaved like targets in miR‐gain and loss‐of‐function experiments described above. Using pmirGLO luciferase, we identified MAP3K4 and HIF as direct targets of miR199. Finally, we generated a miRNA knockdown cassette to reduce expression of Hif1α and Map3k4. Blocking HIF1α resulted in a large reduction of Shh expression in the FEZ, and resulted in severe craniofacial malformations, while blocking Map3k4 altered facial morphology, resulting in altered mediolateral beak development at later stages. Overall, our results demonstrate that Shh signaling within the brain contributes to facial development by regulating a family of miRNAs that then regulate the expression of Shh in the FEZ via targeting the transcription factor Hif1α.

Support or Funding Information

This work was supported by the National Institutes of Health (R01 DE018234‐A1)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Craniofacial Bone Mineral Density in Mice with Osteogenesis Imperfecta (OI)
Authors: McBride Alexandra H , Ladd Summer H , Organ Jason M , Menegaz Rachel A
Published: June 2021

Osteogenesis Imperfecta (OI) is a rare genetic disorder characterized by a point mutation in COL1A1 or COL1A2 genes that leads to the abnormal synthesis and assembly of type I collagen, a major organic component of bone. Clinical manifestations of the severe OI Type III include small body size, limb deformities, and low bone mineral density (BMD) within the post‐cranial skeleton. OI type III often co‐occurs with dentinogenesis imperfecta (DI), especially in juveniles. The goals of this study are: (1) to investigate the relationship between the organic and inorganic components of bone in the craniofacial skeleton; and (2) to examine whether an abnormality in type I collagen, as seen in OI, affects BMD within the craniofacial skeleton.

The homozygous recessive murine mouse (OIM−/−) is a model for OI Type III. Similar to human OI patients, OIM−/− mice exhibit low BMD within post‐cranial elements such as the femur, smaller body size, and DI. OIM−/− mice and WT littermates were weaned at 21 days and CT scans were performed longitudinally at 4 weeks, 10 weeks, and 16 weeks of age. BMD at three regions of interest (ROIs) within the mandible were measured using Bruker CT Analyzer software. ROIs were selected on the basis of high vs. low loading during mastication, and included the temporomandibular joint (TMJ), buccal side of the mandibular corpus at the level of the second molar (M2), and the mandibular symphysis. Kruskal‐Wallis ANOVAs were used to statistically compare BMD between genotypes (α = 0.05). Adult OIM−/− mice (n=6) have significantly lower BMD within the mandibular corpus near M2 as compared to WT mice (n=6). Differences in BMD means between OIM−/− and WT mice were not significant at the TMJ or the mandibular symphysis. Future studies will quantify BMD at additional ROIs within the craniofacial skeleton, and following the ontogenetic trajectory of BMD in the craniofacial skeleton from weaning to adulthood.

Support or Funding Information

This research was supported by an Indiana University Collaborative Research Grant and the Ralph W. and Grace Showalter Trust.

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 homeodomain protein Cux1 is regulated by the Notch signaling pathway in the developing kidney
Authors: Holthaus Conner , Kumar Emmanuel , Carlton Carol , Heuvel Greg Vanden
Published: June 2021

Cux1 is the murine homologue of the Drosophila gene Cut. In Drosophila, multiple genetic interactions between Cut and the Notch and Wingless signaling pathways occur during development. Cux1 is upregulated in rat kidney epithelial cells expressing a constitutively active Notch1 (Notch1ic), and interacts with the groucho homologue Grg4 to regulate p27 expression in the developing kidney. Moreover, Cux1 co‐localizes with several components of the Notch signaling pathway during kidney development. To further evaluate whether Cux1 is an effector of the Notch signaling pathway we used the γ‐secretase inhibitor DAPT to block all Notch signaling in kidney organ cultures. DAPT treatment significantly inhibited growth of wild‐type metanephroi. In contrast, Cux1 transgenic kidney cultures treated with DAPT were not growth inhibited, but showed high levels of cell proliferation in the nephrogenic zone, and ureteric bud branching was similar to vehicle treated kidney cultures. Notch1ic cells showed an increase in cell proliferation compared to wild type RKE cells, however, antisense knockdown of Cux1 in the Notch1ic cells reduced cell proliferation to the same level as wild type cells. Finally, the targeted deletion of RBPJ/k, a transcription factor required for canonical Notch signaling, in the developing ureteric bud and collecting duct, resulted in the complete absence of Cux1 protein specifically in the ureteric bud and collecting duct cells. Taken together, these results suggest that Notch signaling regulates the expression of Cux1 in the developing kidney, and that this pathway is conserved across species.

Support or Funding Information

NIH DK58377, WMED Research Funds

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In Utero Exposure to Nicotine Causes Post‐natal Anemia
Authors: Durham Emily L , Howie R Nicole , Grey Zachary , Hall SarahRose , Larson Nicholas , Patterson Kimberly , Warren Graham , LaRue Amanda C , Cray James
Published: June 2021

Almost ¼ of the world population suffers from anemia, a condition characterized by an insufficient number of red blood cells or oxygen‐carrying capacity within the blood. Anemia is associated with fatigue, weakness, and dizziness, and when it occurs early in life it may alter the course of behavioral and cognitive development in human and murine populations causing decreased physical activity and responsiveness. Pregnant women and children are particularly vulnerable to anemia as are individuals who use nicotine. Despite the link between adverse birth outcomes of pre‐ and perinatal nicotine exposure, 11% of US women continue smoking or using alternative nicotine products throughout pregnancy. We hypothesize that in utero exposure to nicotine will result in post‐natal anemia precipitating a characteristic abnormal bone phenotype. To test this hypothesis, wild type C57BL6 males and females were utilized to produce in utero nicotine exposed litters. Drinking water for pregnant dams was supplemented with nicotine. Pups were grown to 15 days postnatal then sacrificed. Blood and whole skulls were collected for complete blood count, micro‐computed tomography (μCT), and histological analyses. Complete blood count data comparing nicotine exposed pups to unexposed controls indicated increased mean corpuscular volume indicative of macrocytic anemia in nicotine exposed mice. Additionally, nicotine exposed pups presented with low platelet counts also associated with anemia. μCT analysis of bone surface compared to bone volume indicated a greater ratio in individuals exposed to nicotine, which suggests weaker pitted bone also associated with anemia. Histological investigation of osteoclast activity indicated greater osteoclast activity in individuals exposed to nicotine. Together, these data indicate that exposure to nicotine in utero precipitates a negative bone phenotype associated with nutritional deficiency and anemia.

Support or Funding Information

This research was supported by the National Institutes of Health (NIH) National Institute of Dental and Craniofacial Research (NIDCR) [R03DE026192, 5T32DE017551 F31DE026684], National Center for Advancing Translational Sciences [UL1TR000062], and Plastic Surgery Foundation [Pilot Award 512114]. This study utilized the facilities and resources of the Medical University of South Carolina Center for Oral Health Research supported by the NIH/NIGM [P30GM103331].

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterizing the Expression Pattern and Requirement of Sf3b4 During Mouse Embryonic Development
Authors: Kumar Shruti , Alam Sabrina , Chan Wesley , Beauchamp Marie‐Claude , Jerome‐Majewska Lodyie
Published: June 2021

Nager Syndrome (NS) is a rare disorder that affects the face as well as the limb including both hands and feet. Patients with NS typically have malar and mandibular hypoplasia, cleft palate, as well as hearing problems. Limb defects include radial hypoplasia as well as thumb abnormalities. Using exome sequencing, NS was attributed to haploinsufficiency of the SF3B4 gene, an important component of the U2 subunit of the spliceosome complex. We hypothesized that Sf3b4 will show tissue‐specific expression during development and that mice with heterozygous mutation in this gene will model NS. To create heterozygous Sf3b4 mutant mice, we used CRISPR/Cas9 to target loxP sequences in intronic regions flanking exon 2 and 3 of the Sf3b4 gene. We will breed loxP founders with Wnt‐1 Cre transgenic mice, and report the resulting phenotypes. Additionally, whole mount in situ hybridization was used to examine expression of Sf3b4 during embryonic development of wild type mouse. We found that Sf3b4 shows ubiquitous expression at early stages of development. In midgestation embryos, although still globally expressed, Sf3b4 expression becomes stronger in the maxillomandibular region, limbs and tail bud. Our research will elucidate the expression of Sf3b4 in embryonic tissues affected in NS patients and generate a mouse model that will be used to further characterize the molecular basis of this syndrome. As there are currently no mouse models or therapy available for NS our work will help to identify some of the targets of Sf3b4 and pave the path for the creation of new therapies for spliceosomal disorders.

Support or Funding Information

Canadian Institutes of 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.
Identification of nutrient metabolites capable of altering the epigenetic status at specific loci
Authors: Hayakawa Koji , Nishitani Kenta , Tanaka Satoshi
Published: June 2021

Nutrients can influence cellular functions associated with physiological and pathological processes. Major energy sources in our body, such as glucose and amino acids, are decomposed, resynthesized via numerous synthetic pathways, and finally, metabolized into energy molecules (e.g., ATP) and metabolites involved in various molecular events such as gene expression. Our previous study demonstrates that a sugar metabolite, N‐acetyl mannosamine, when used as a supplement in the culture medium, can induce the differentiation of pluripotent stem cells into orexin neurons and activate the epigenetic status at the orexin‐coding gene locus. Thus, our observations suggest that some metabolites can change the fate of a cell by participating in epigenetic alterations at specific loci. However, there is insufficient information about the role of specific nutrients and their metabolites in altering the epigenetic status. In the present study, we aimed to screen for metabolites that function as epigenetic modulators. We supplemented the culture medium with 129 metabolites individually during the induction of neural differentiation from human iPS cells (hiPSCs). We investigated the epigenetic status at 40 loci of the neural peptide‐coding genes by ChIP‐qPCR, using a high‐throughput platform. Trimethylation of lysine 4 of histone H3 (H3K4me3) and histone O‐GlcNAcylation of serine 40 of H2A (H2AS40Gc) were used as active epigenetic markers. Metabolites 4 and 12 considerably altered the levels of H3K4me3 and H2AS40Gc, respectively, at the loci of most genes, except at those of genes encoding pan‐neural markers. Among them, metabolites produced via the kynurenine pathway (one of the tryptophan metabolic pathways) increased H3K4me3 and H2AS40Gc levels at multiple neural peptide‐encoding gene loci such as CART (cocaine‐and amphetamine‐regulated transcript) and OXT (oxytocin). The mRNA expression levels of these genes were also found to have increased following supplementation of the culture medium with metabolites of the kynurenine pathway. In other words, supplementation with these metabolites could effectively induce the differentiation of hiPSCs into CART‐ and OXT‐producing neurons. Collectively, we have identified metabolites that can alter the epigenetic status at specific loci. Our findings provide a better understanding of the use of nutrients or bioactive food components in modifying epigenetic mechanisms.

Support or Funding Information

This work was supported by Lotte Shigemitsu Prize.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis of Chromatin Accessibility Landscapes in Esophageal Squamous Cell Carcinoma
Authors: Choi Sangyong , Sathe Adwait , Mathe Ewy , Xing Chao , Pan Zui
Published: June 2021

Esophageal cancer ranks sixth among all cancers in mortality and is associated with malfunction in gene regulation and expression. Assay for Transposase‐Accessible Chromatin with high‐throughput sequencing (ATAC‐seq) can reveal the locations of regulatory elements across the genome and discover the mechanisms of chromatin structure that govern gene expression programs. This study compared the chromatin accessibility landscapes between esophageal squamous cell carcinoma cell lines (ESCC), i.e. KYSE‐30 and KYSE‐150, and non‐cancerous esophageal epithelial cell lines (NC), i.e. HET‐1A and NES‐G4T. Opened and closed chromatin regions were associated to the nearest genes, and gene ontology analyses (GO) were performed. The results demonstrated that many regions were more accessible in ESCC cells than in NC cells. These regions belong to the GO terms including positive regulation of GTPase activity, metabolic process, and nucleosome assembly. Furthermore, the promoter‐proximal peaks at genome‐wide scale predicted that several genes in histone gene cluster 1 on chromosome 6p22 were transcriptionally active in ESCC cells. To confirm that the promoter region with enhanced accessibility indeed is indicative to gene expression, the mRNA level of a few representative genes were examined. For example, the expression of PLCB1 gene, which encodes phospholipase C beta 1, a key regulator for GTPase activity and MAPK signaling, was confirmed to be up‐regulated in both ESCC cells. There were also some regions more inaccessible in ESCC cells than in NC cells and they belonged to different GO terms, chiefly associated with cellular homeostasis. This study may aid to reveal the instrumental epigenetic changes responsible for differential gene expression and ESCC development.

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 Histone Acetyltransferase GCN5 and Transcriptional Coactivator ADA2b Affect Trichome Initiation in Arabidopsis thaliana
Authors: Hark Amy T. , Caiola Hanna , Sockler Patrick , Kotak Jenna , Kendig Ashley , Cann Kelly , McCain Elizabeth R.
Published: June 2021

While genome sequencing projects have helped us uncover what information is encoded in DNA of many species, one critical challenge that remains is to understand how gene function is controlled. We investigate the roles of chromatin‐modifying factors in regulating developmental processes in Arabidopsis thaliana. Our current work explores how the histone acetyltransferase GCN5 and the associated transcriptional adaptor ADA2b act to define which leaf cells are selected to become trichomes, single‐celled protrusions implicated in defense mechanisms. In a loss‐of‐function ada2b mutant background, trichome number on rosette leaves is decreased while in gcn5 mutant backgrounds the number of trichomes tends to increase. Using qRT‐PCR analysis, we have established a role for GCN5 in impacting the expression of key players in the trichome initiation pathway including the paralogs Repressor of GA (RGA) and GA Insensitive (GAI). Gibberellic acid (GA) signaling stimulates trichome initiation by blocking the DELLA repressors RGA and GAI, which in turn block a transcription factor cascade that controls trichome development. This approach has the potential to identify novel connections between GA, a critical plant hormone, and histone acetylation in plant growth. It also provides an example of how chromatin modifiers and transcription factors interact to drive developmental processes.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Genome Wide Microarray Profiling Identified Differentially Expressed Circulating Biomarkers Associated with Early Development of Diabetic Cardiomyopathy
Authors: PANT TARUN , DHANASEKARAN ANURADHA , BAI XIAOWEN , BOSNJAK ZELJKO , GE ZHI‐DONG
Published: June 2021

Introduction

Circulating long noncoding RNAs (lncRNAs) are emerging as valuable biomarkers in numerous diseases. However the expression signature of circulating lncRNAs is yet to be identified in diabetic cardiomyopathy (DCM). We used computational and system biology approach to predict circulating lncRNAs using plasma RNA before and after DCM.

Hypothesis

We hypothesized that lncRNAs could serve as blood‐based prognostic and diagnostic biomarkers in DCM detection.

Methods

The left ventricular function was measured with echocardiography in control and db/db mice. Expression profiling of lncRNAs was performed using LncRNA Array v3.0 (8 × 60K, Arraystar).

Results

A total of 17,124 lncRNAs and 13,333 mRNAs were profiled of which 814 lncRNAs and 2707 mRNAs were deregulated, and more than half were intergenic lncRNAs and antisense lncRNAs. Ten dysregulated plasma lncRNAs were validated by quantitative PCR assays. For the gene ontologies related to plasma, the enrichment was observed for those mRNAs which were found to be significantly co‐expressed with the neighboring lncRNAs during DCM. Bioinformatics analysis revealed that the altered lncRNAs regulated the differentially expressed mRNAs which were profoundly enriched for pathways such as insulin signaling, fatty acid metabolism, MAPK signaling, and hypertrophy cardiomyopathy which in the past has been associated with diabetic pathogenesis. To evaluate the translational efficiency of our mouse model, we experimentally validated the results using human induced pluripotent cell‐derived cardiomyocytes (hiPSCs‐CMs) for the in vitro DCM model. We found that most of the gene expression patterns were repeated in hiPSCs‐CMs and several of these lncRNAs genes have been associated with DCM in the past.

Conclusion

Circulating lncRNAs are dysregulated in DCM, and our results depict the changing transcriptome as a function of disease progression and functional role of lncRNAs. These lncRNAs may serve as promising targets for DCM prevention, early diagnosis, and therapy.

Support or Funding Information

This work was supported, in part, by a National Institutes of Health research grant P01GM 066730 (to Dr. Bosnjak) from the United States Public Health Services, Bethesda, Maryland, USA. The funding body had no role in the design of the study, collection, analysis, and interpretation of data, and in writing the abstract.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Alcohol induces TGF‐beta through suppression of miR‐1946a
Authors: Sueblinvong Viranuj , Mills Stephen T , Fan Xian
Published: June 2021

Rationale

We previously showed that chronic alcohol ingestion promotes fibroproliferative disrepair following bleomycin‐induced acute lung injury in a mouse model and that is associated with exaggerated constitutive and injury‐provoked expression of TGFβ. Importantly, the aberrant expression of TGFβ has been identified as a proximal event that mediates the ‘alcoholic lung’ phenotype. However, the mechanisms by which alcohol induces TGFβ1 has not been identified. Several microRNAs (miR) have been shown to directly target TGFβ. Using an in silico analysis, we identified that miR‐1946a is predicted to directly bind to the 3′ untranslated region (UTR) of the TGFβ mRNA and thereby inhibit its transcription. We hypothesize that alcohol‐mediated miR‐1946a leading to an induction of TGFβ expression.

Methods

Mouse primary lungfibroblasts (PLFs) were cultured ± alcohol (60 mM) for 24–48 hrs, miR‐1946aexpression was quantified. In parallel, PLFs were cultured with miR‐1946amimic, miR‐1946a inhibitor, or appropriate controls in the presence of absent of alcohol, then assessed for TGFβ gene and protein expression. Lastly, to further comfirm the miR‐1946abinding site on TGFβ 3′UTR, PLFs were transfected with vector containing TGFβ3′UTR (or control clone) tagged with luciferase reporter and miR‐1946a mimic(or negative mimic), then analyzed for luciferase activity.

Results

Alcohol suppressed PLFs expression of miR‐1946a. Treatment of PLFs with miR‐1946a mimic suppressed alcohol‐induced TGFβ expression while miR‐1946a inhibitor induced TGFβ expression. Lastly, miR‐1946a mimic suppressed TGFβ 3′UTR luciferase activity.

Conclusion

These data suggest that miR‐1946a modulates TGFβ expression likely through direct interaction with TGFβ 3′UTR. These findings identify a novel mechanism by which alcohol induces TGFβ in the lung which would help us design therapeutic or preventative tools for alcoholic patients with or at risk of acute lung injury.

Support or Funding Information

NIH K08 AA021404‐01

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Measuring the Stability of RNA5.0 in Human Cytomegalovirus (HCMV)
Authors: Tsingine Hannah P , Kulesza Caroline
Published: June 2021

Human Cytomegalovirus (HCMV) is a member of the beta herpesvirus family. HCMV establishes a lifelong latent infection in its host, and although it infects a variety of cell types and replicates persistently, it has an asymptomatic effect in healthy humans. Primary or reactivated HCMV infections have the ability to be transferred via bodily secretions such as saliva, urine, or breast milk, and through organ transplants. Unfortunate organ recipients who contract the virus may suffer from pneumonia and/or acute organ rejection. HCMV is the largest of the human herpesviruses containing a DNA genome of approximately 240 kilobases encoding 225 open reading frames (ORFs). It's large and complex genome also encodes long noncoding RNAs (lncRNA) which are known for not being translated, but rather function as RNA. In HCMV, one of the lncRNA, known as RNA5.0, is a stable intron that is believed to play an important role in viral pathogenesis. Recent studies have shown that RNA5.0 is a nuclear localized stable intron that is transcribed by RNA polymerase II, and that it also consists of high adenine and thymine sequences. The true role of RNA5.0 is still unknown, so it could activate transcription, regulate gene silencing, or play a role in HCMV latency. To begin to understand why an intron that should be degraded within minutes is stable for hours, we will measure the half‐lives of viral noncoding RNAs using transfected cultured 293T cells.

Support or Funding Information

NIH MARC T34GM092711

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Micro‐RNA isolated from liquid biopsies serve as novel molecular biomarkers for congestive heart failure in dogs
Authors: Shands Kristal , Ewing Carl , Ewing Coralis , Bohan Amy
Published: June 2021

Objective

To characterize expression profiles of circulating microRNAs via miRNA isolation from plasma (Qiagen) for dogs with congestive heart failure (CHF) secondary to myxomatous mitral valve degeneration (MMVD).

Materials and Methods

9 healthy small breed client‐owned dogs and 9 small breed client‐owned dogs (ages 9 and up) with CHF secondary to MMVD. Blood samples were collected from both healthy and affected dogs. Isolated microRNAs from plasma were subjected to quantitative reverse transcription PCR (qRT‐PCR) used to examine expression profiles of differentially expressed circulating microRNAs of dogs with CHF.

Results

Results of qRT‐PCR assays confirmed up‐regulation of several microRNAs as well as downregulation of multiple microRNAs. Results of qRT‐PCR assays were highly correlated with NGS data from a prior study done and supported our results.

Conclusion

These results suggested that miRNAs serve as novel biomarkers for congestive heart failure (CHF) in small breed canines.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
miRNAs regulate the expression of sterol‐O acyltransferase 1 through inhibiting the 3′UTR of TGFβ receptor type 1 during embryonic development
Authors: LIN HAN‐JEN , DING SHIH‐TORNG
Published: June 2021

Nutrients are utilized and re‐constructed by endodermal epithelial cells (EECs) of yolk sac membrane (YSM) in avian species during embryonic development. Sterol‐O acyltransferase 1 (SOAT1) is the key enzyme to convert cholesterol to cholesteryl ester, and for packaging into very‐low‐density lipoproteins for further delivering to growing embryos. During embryonic development, the quick absorption of yolk is matched with the significant changes of SOAT1 mRNA and enzymatic activity. MicroRNAs (miRNAs), groups of highly conserved small non‐coding RNA molecules, are usually ~23 nucleotides across species, serving classical biofunctions as mRNA‐targeting cleavage, translation repression and mRNA deadenylation. miRNAs have been identified to regulate angiogenesis, invasion and metabolism during mammalian development. However, the involvement and importance of miRNAs in lipid utilization during avian embryogenesis remains unclear.

Using a RNA sequencing technique, we found many miRNAs expressed during quail embryonic development. We confirmed the expression pattern with real time PCR and several miRNAs (gga‐miR‐181a‐5p, gga‐miR‐133a‐5p, and gga‐miR‐429‐3p) were selected because they are candidates targeting to the receptor (TGFβ receptor type 1, TGFBR1) that regulates the expression of SOAT1. Similar to SOAT1 mRNA concentration, the gga‐miR‐181a‐5p expressions were gradually elevated during development, but gga‐miR‐429‐3p were presented in the opposite direction. Moreover, transfection with gga‐miR‐181a‐5p, gga‐133a‐5p, or gga‐429‐3p inhibited TGFBR1 and SOAT1 mRNA expressions in EECs. The 3′ untranslated region (3′UTR) of TGFBR1 is speculated to be one of the targets of miRNAs (gga‐miR‐181a‐5p, gga‐miR‐133a‐5p, and gga‐miR‐429‐3p).

Taken together, miRNAs expression during embryonic development can regulate SOAT1 expression through inhibiting the 3′UTR of TGFBR1. This is indicative of possible regulation of avian yolk lipid utilization and modification of hatchability by modifying miRNA expression.

Support or Funding Information

Ministry of Science and Technology, Taiwan (ROC)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rapid RNA‐Protein Interaction Detection with RNA5.0 of HCMV
Authors: Mickens Kassandra , Mickens Kaylee , Kulesza Caroline
Published: June 2021

CMV is the most common cause of both congenital infection and post‐transplant infection worldwide. CMV infection can cause severe symptoms: pneumonia, encephalitis, and inflammation of vital organs. There is currently no vaccine for CMV. An understanding of how CMV progresses from its acute phase to persistence would illuminate features of the pathogen that could lead to novel treatments for the disease which block the virus from reaching latency.

RNA5.0 is a lncRNA produced during CMV infection. Studies using the homologous RNA7.2 a mouse model has shown when the RNA is not expressed in vivo, CMV infection cannot reach its persistent phase. However, how it performs this function remains unclear. Our study hopes to help the field to move forward by expanding the limited knowledge about RNA5.0's function in viral persistence at a molecular level by purifying and identifying the proteins. We propose identifying proteins that bind to RNA5.0 with known cellular functions will allow us to narrow the speculation surrounding RNA5.0's function, as their functions are likely related.

To purify these proteins, we will utilize the RaPID (RNA–protein interaction detection) method outlined in Ramanathan et al. 2018. In this novel approach, RNA stem loops flank RNA sequence of interest and recruits the RaPID fusion protein that biotinylates proteins proximal to inserted RNA. A streptavidin pulldown is used to capture biotinylated proteins for analysis by mass spectrometry (MS). Because the RaPID method has limited reach, in order to characterize all the proteins that bind to the 5kb lncRNA of CMV we will use 50 overlapping 150bp segments with 50bp overlaps.

Support or Funding Information

Fort Lewis College Undergraduate Research, Scholarship, and Creative Activity 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.