bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anatomy of the Proboscideal Nerve in the Proboscis of the African Elephant (Loxodonta africana)
Authors: Nabavizadeh Ali , Reidenberg Joy S. 
Published: June 2021

The elephant proboscis (i.e., trunk), a unique muscular organ formed by the combined narial and upper lip region, is a complex muscular hydrostat adapted for a multitude of functions, including grasping, holding, and transporting objects, siphoning, holding, and spraying water, and communication. The proboscis has previously been described as containing tens of thousands of muscle fascicles, each working individually to help bend, curve, and curl the trunk in all directions at any point along its length. Although the muscular system of the proboscis has previously been described in some depth, the unique nervous innervation of the proboscis has been given minimal attention. Here, the previously frozen head of an adult female African elephant (Loxodonta africana) is dissected to investigate the anatomy of the neuromuscular anatomy of the proboscis in more depth. In addition to the complexity of interlacing muscle fascicles and the integrated connective tissues, we identified the proboscideal nerve, a large, 2.5 to 3 centimeter thick nerve that is unique to elephants. The proboscideal nerve is made up of the combined distal nerve fibers of the infraorbital branch of the maxillary nerve (CN V3) and zygomatic branch of the facial nerve (CN VII). Specifically, as the infraorbital branch of the maxillary nerve (carrying sensory innervation) exits the cranium through the infraorbital foramen, the zygomatic branch of the facial nerve (carrying motor innervation) runs along the ventral border of the zygomatic arch and joins the maxillary nerve rostrodorsal to the masseter muscle to form the proboscideal nerve. The proboscideal nerve then travels down the length of the proboscis transmitting small sensory and motor nerve fibers to all muscle fascicles and surrounding skin and tissue. Given this anatomy, fine‐tuned neuromuscular control and coordination of the individual nerve fibers are necessary to control the diverse movements that occur throughout the length of the proboscis. This specialized evolutionary adaptation helped shape a functionally innovative organ that grew, modified, and persisted throughout the course of proboscidean evolution.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 Radiographic Analysis of Phalanx Shapes
Authors: Neiswonger Madison Paige , Hartnell Layne S. , Bender Emily M. , Zdilla Matthew J. 
Published: June 2021

Understanding differences in the characteristics of human phalanxes is important for varied disciplines including anthropology, forensics, and surgery. For the most part, analysis of the phalanxes has been limited to “traditional” morphometric measurements such as length. Therefore, this preliminary study sought to utilize geometric morphometric methods in the analysis of phalanxes. After exclusions, a total of 1830 phalanxes (618 proximal, 610 middle, and 602 distal) from digits 2–4 were analyzed from PA radiographs. A set of four landmark points were applied to each phalanx. Canonical variate analysis (CVA) was performed among phalanx rows (i.e., proximal, middle, and distal). Additionally, CVA was utilized to identify shape variance of phalanxes, within‐row, according to digit number (i.e., 2nd, 3rd, 4th, 5th phalanxes within a given row). The results of this research suggest that geometric morphometric analysis of phalanxes may aid in the differentiation and identification of phalanxes.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differentiation of neuronal population in cytoarchitecture of spinal cord using Modified Thionin Staining
Authors: Potaliya Pushpa , Ghatak Surajit 
Published: June 2021

BackgroundNumerous staining techniques for neuronal tissue have an inherent unreliability even after elaborate procedures. A non‐methylated thiazin dye, thionin, has been extensively used in the past for neuronal studies. However procedure is not suitable for paraffin embedded tissue. In this study a modification of thionin stain was done for paraffin embedded neuronal tissue.AimFor better differential staining of neuronal tissue in spinal cord with modified thionin staining.Material and methodThe formalin fixed material was stained by conventional H & E, thionin and a modified thionin method to evaluate staining results. For this spinal cord sections were treated and stained with acid fuschin, oxalic acid and thionin.ResultThe procedure imparted a distinct differential coloration of myelin and neurons. Myelinated fibres appear pinkish whereas nerve cell bodies and glial nuclei were stained blue.ConclusionThe modified thionin method was found to be a reliable mean to provide satisfactory results with neuronal tissue in spinal cord.Support or Funding InformationNilThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cell Based Therapy Reduces Secondary Damage and Increases Extent of Microglial Activation Following Cortical Injury
Authors: Orczykowski Mary E , Calderazzo Samantha M , Shobin Eli , Pessina Monica A , Oblak Adrian L , Finklestein Seth P , Kramer Brian C , Mortazavi Farzad , Rosene Douglas L , Moore Tara L 
Published: June 2021

Cortical injury elicits long‐term cytotoxic and cytoprotective mechanisms within the brain and the balance of these pathways can determine the functional outcome for the individual. Cytotoxicity is exacerbated by production of reactive oxygen species, accumulation of iron, and peroxidation of cell membranes and myelin. There are currently no neurorestorative treatments to aid in balancing the cytotoxic and cytoprotective mechanisms following cortical injury. Cell based therapies are an emerging treatment that may function in immunomodulation, reduction of secondary damage, and reorganization of surviving structures. We previously evaluated human umbilical tissue‐derived cells (hUTC) in our non‐human primate model of cortical injury restricted to the hand area of primary motor cortex. Systemic hUTC treatment resulted in significantly greater recovery of fine motor function compared to vehicle controls. Here we investigate the hypothesis that hUTC treatment reduces oxidative damage and iron accumulation and increases the extent of the microglial response to cortical injury. To test this, brain sections from these monkeys were processed using immunohistochemistry to quantify oxidative damage (4‐HNE) and activated microglia (LN3), and Prussian Blue to quantify iron. hUTC treated subjects exhibited significantly reduced oxidative damage in the sublesional white matter and iron accumulation in the perilesional area as well as a significant increase in activated microglia along downstream white matter. Increased perilesional iron accumulation was associated with greater perilesional oxidative damage and larger reconstructed lesion volume. These findings support the hypothesis that systemic hUTC administered 24 hours after cortical damage decreases the cytotoxic response while increasing the extent of microglial activation.Support or Funding InformationThis study was supported by a contract from Advanced Technologies and Regenerative Medicine (ATRM), LLC. [RR# 101115‐PR] who provided the cell therapy product and the vehicle control and by the National Institutes of Health [NIH‐NINDS R21NS081261].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 CAFFEINE REDUCES DNA METHYLATION IN HUMAN MESENCHYMAL STEM CELLS
Authors: Pregmon Emily , Seat Mara , Poulsen Ryan , Wendler Christopher Charles 
Published: June 2021

There is growing evidence that alterations in the intrauterine environment can have lasting effects on an individual's health. By disrupting prenatal development, environmental factors can lead to the fetal programming of adult disease, including cardiovascular disease. One substance that fetuses are frequently exposed to is caffeine, which is a non‐selective adenosine receptor antagonist. Caffeine consumption during the first trimester of pregnancy is reported by 60% of pregnant women. Human studies have shown correlations between caffeine use during pregnancy and low birth weight, childhood obesity, and neurodevelopmental deficits in children. Although, all do not accept some of these deleterious effects, it is concerning enough that the United States and many Nordic countries recommend that pregnant women consume less than 200 mg/day, or about 2 cups of coffee a day.Our lab has demonstrated that in utero caffeine exposure in mice produces defects in cardiac gene expression, morphology, and function in adulthood. Furthermore, we have shown that in utero caffeine exposure alters DNA methylation patterns in adult hearts. Based on these data, we hypothesize that in utero caffeine exposure leads to altered DNA methylation that is maintained into adulthood leading to altered gene expression and reduced heart function in adults.It is difficult to study the effects of caffeine on human development at the molecular level. Therefore, we have chosen to examine human mesenchymal stem cells (MSCs) obtained from umbilical cords of infants with known caffeine exposure during development. These cells were obtained from Dr. Kristen Boyle of The Healthy Start Project at the University of Colorado School of Medicine. We obtained a subset of these cell lines that were divided into two groups based on caffeine exposure during pregnancy.Initially, we isolated RNA and DNA from these MSC lines to compare gene expression and DNA methylation between low and high caffeine exposure groups. A global DNA methylation assay revealed that the high caffeine group had a 29.98% decrease in global DNA methylation compared to the low caffeine group (p ≤ 0.01, N = 3). Furthermore, qPCR revealed a 3.5 fold decrease in DNA methyltransferase 3a (DNMT3a) expression in the high caffeine exposure group. DNMT3a is one of three enzymes, including DNMT1 and DNMT3b, responsible for DNA methylation during embryonic development.In summary, our initial results indicate a correlation between high caffeine exposure and reduced global DNA methylation within human MSCs obtained from umbilical cords with known caffeine exposure. In addition, we observed altered gene expression in MSCs exposed to high caffeine levels in utero.Support or Funding InformationChildren's Miracle Networkat the University of FloridaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Lethal Replication Roadblock Imposed by a Catalytically Inactive DNA Polymerase
Authors: Hernandez Alfredo J , Lee Seung‐Joo , Chan Seungwoo , Lee Jaehun , Loparo Joseph , Richardson Charles C 
Published: June 2021

Bacteriophage T7 encodes its own DNA polymerase, the product of gene 5. In isolation, gene product 5 (gp5) is a DNA polymerase of low processivity, however, it becomes highly processive upon formation of a complex with E. coli thioredoxin. In the course of investigating the transfer of primers from T7 DNA primase to T7 DNA polymerase, produced a gp5 variant lacking polymerase activity. Expression of a gp5 variant where the metal binding site in the polymerase domain was inactivated was found to be highly toxic to E. coli. This toxicity was strictly dependent on the presence of a functional E. coli trxA allele and T7 RNA polymerase‐driven expression. The closely related Klenow fragment of DNA polymerase I containing an engineered gp5 thioredoxin‐binding domain did not show any toxicity. Purified polymerase‐inactive gp5 drastically inhibited the E. coli replisome in vitro, but only in the presence of thioredoxin. Mutation of two critical metal binding site residues promote the formation of long‐lived protein‐DNA complexes that may block the displacement of the E. coli replication machinery. On the other hand, the inactive gp5 mutant only partially inhibits the T7 replisome, T7 DNA polymerase, or Klenow fragment. We envision that this T7 gp5 allele, in combination with a functional copy of TrxA, can be used a genetic building block in the design of synthetic biology approaches as a cell‐growth shutoff mechanism or a timed protein antibiotic.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exploring a potential link between cell cycle regulation and Cancer Testis gene misregulation
Authors: Alhewati Sabha Salem , Goetsch Paul 
Published: June 2021

Oncogenesis and misexpression of germline genes like Cancer Testis (CT) genes, called the soma‐to‐germline transformation, are commonly linked, but the mechanisms that drive the phenomenon remain underexplored. In the model system Caenorhabditis elegans, a soma‐to‐germline transformation occurs when the function of the highly conserved DREAM complex (for Dp, Retinoblastoma(Rb)‐like, E2F, and MuvB) is attenuated. In mammalian cells, DREAM's activity overlaps with the Retinoblastoma(Rb) protein to mediate establishment of cell cycle exit into quiescence, or G0. We hypothesize that CT gene misexpression in cancer cells occurs following loss of DREAM and Rb activity, similar to the soma‐to‐germline transformations that occur in C. elegans. Therefore, we expect that cancer cell lines that express CT genes will be unable to maintain an effective cell cycle exit. To test our hypothesis, cell cycle distributions of 8 cancer cell lines were analyzed using propidium iodide (PI) staining and flow cytometry before and after serum starvation. We observed that only 3 of the cell lines failed to arrest under growth limiting conditions, with the other 5 exhibiting some enrichment in G0/G1, indicating that these cell lines that did not arrest might exhibit upregulated expression of CT genes, and DREAM complex and Rb are deregulated. However, the cell lines that we observe G0arrest may still be dysfunctional in either DREAM complex or Rb activity. Therefore, we will next test expression analysis will test whether CT genes are differentially regulated between these groups and whether DREAM and/or Rb regulation is attenuated. This study is the first step towards understanding whether cell cycle regulation and the soma‐to‐germline transformation are linked in mammalian cells.Support or Funding InformationThe Saudi Arabian Cultural Mission (SACM)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein‐DNA interactomes of cardiac transcription factors GATA4 and NKX2‐5
Authors: Carrasquillo Emmanuel , Rodriguez Jose Arcadio 
Published: June 2021

Every year over 100,000 new cases of Congenital Heart Disease (CHD) are reported, being one of the most common disease in neonates. CHDs are characterized by anomalies in the heart and its great vessels during development and physiology. Transcription Factors (TFs) such as GATA4 and NKX2‐5 play a vital role in the regulation of genes involved in human heart development and functionality. Identified point mutations in the DNA‐binding domain (DBD) of GATA4 and NKX2‐5 are among multiple mutations associated to CHD. We hypothesize that non‐synonymous mutations in the DBD of GATA4 and NKX2‐5 will alter their DNA‐binding properties ultimately affecting the whole transcription pathway for a proper heart development. To address this hypothesis, we want to study how the GATA4 and NKX2‐5 DNA‐protein interactomes are affected by the non‐synonymous mutations found in CHD patients. We have successfully cloned the DNA binding domain (DBD) of GATA4 and NKX2‐5 via Gibson‐assembly. Both of which were over‐expressed and purified. Functional analysis protocols using electrophoretic mobility shift assay (EMSA) confirms protein binding to its DNA binding sites and therefore functional. We will measure the comprehensive protein‐DNA interactomes of missense mutants of GATA4 and NKX2‐5 using High‐Throughput Systematic Evolution of Ligands by Exponential Enrichment (HT‐SELEX), and use bioinformatics tools to predict changes in gene targets between wild‐type and mutant TFs.Support or Funding InformationThis research was supported by:1) Research Initiative for Scientific Enhancement (RISE) Grant #: 5R25GM061151‐172) NIH SC1GM127231This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Methylation of RB1 CpG island by SUV39H1/DNMT3A promotes malignant melanoma development
Authors: Kim Garam , Choi Hong Seok 
Published: June 2021

Melanoma is among the most aggressive and treatment‐resistant human cancers. Aberrant histone H3 methylation at Lys 9 (H3K9) correlates with carcinogenic gene silencing but the significance of suppressor of variegation 3–9 homolog 1 (SUV39H1), a H3K9‐specific methyltransferase, in melanoma initiation and progression remains unclear. Here we show that SUV39H1‐mediated H3K9 trimethylation (H3K9me3) facilitates retinoblastoma (RB1) promoter CpG island methylation by interacting with DNMT3A and decreasing RB mRNA and protein in melanoma cells. Reduced RB abundance, in turn, impairs E2F1 transcriptional inhibition leading to increased peptidyl‐prolyl cis‐trans isomerase NIMA interacting 1 (PIN1) levels, human keratinocyte neoplastic cell transformation, and melanoma tumorigenesis via enhanced RAF1/MEKs/ERKs signaling pathway activation. In a synergistic model with B16‐F1 murine melanoma cells, SUV39H1 and PIN1 overexpression increased melanoma growth, which was abrogated by their inhibition in SUV39H1‐overexpressing B16‐F1 mice. SUV39H1 also positively correlated with PIN1 expression in human melanoma. Our studies establish SUV39H1 as an oncogene in melanoma and underscore the role of chromatin factors in regulating tumorigenesis.Support or Funding InformationThis research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number : HI18C1083).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of αvβ3 Integrin Proteins in Glaucoma
Authors: Fassler Amy 
Published: June 2021

Glaucoma, the leading cause of vision loss worldwide, occurs when elevated intraocular pressure damages the optic nerve. Intraocular pressure increases when there is a blockage of the trabecular meshwork, a drainage system at the base of the cornea. This process is dictated by a bidirectional signalling protein composed of a specific combination non‐covalently bonded α and β integrin subunits called the αvβ3 integrin. In an inactive state, the integrin is is positioned as a switchblade and its cytoplasmic tails are held together by a salt bridge. To become activated, the integrin must extend; the salt bridge must be broken, and the cytoplasmic tails spread apart. During outside‐in signalling, a ligand binds to the αvβ3 integrin which forces it to re‐conform to the active state and results in the transmission of mechanical and chemical cues from the extracellular matrix into the cell by means of a protein cascade. This leads to the trabecular meshwork becoming more dense and interwoven. A possible hypothesis to explain the connection between integrins and glaucoma is that the trabecular meshwork can not turn off the αvβ3 integrin which results in a reduced outflow of the aqueous humor and thus, increased intraocular pressure. Understanding the modulation of intraocular pressure by integrins can lead to therapeutic targets to treat glaucoma. The Marshfield High School MSOE Center for BioMolecular Modelling for SMART Team used 3D Modelling and printing technology to examine structure‐ function relationships of the αvβ3 integrin protein.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of Carbonic Anhydrase Expression in Experimental Colitis: A Possible Mechanistic Link with Na‐H Exchanger
Authors: Khan Islam , Al‐Shamali Amna 
Published: June 2021

In this study we investigated a role and regulation of carbonic anhydrase (CA) and a possible mechanistic link with the NHE expression in experimental colitis. Intra‐rectal instillation of trinitrobenzenesulphonic acid was used to induce colitis in rats. On day 6 post‐TNBS, colon and ileal segments were taken out and used to measure expression levels of the CA‐I, ‐II and ‐IV isoforms by ECL western blot analysis. These isoforms together with NHE‐1 were significantly suppressed in the inflamed colon. Immunoprecipitation and confocal immunofluorescence demonstrated co‐localization of CA‐I and CA‐II, but not the CA‐IV isoform with NHE‐1 in the colon. However, the binding of CA isoforms with NHE‐1 was more suppressed as compared to their expression level in inflamed colon. Only CA‐I was found to interact with an 83 aa long NHE‐1 C‐terminal peptide. Ex‐vivo studies demonstrated suppression of CA‐I and CA‐II isoforms by TNF‐alpha treatment of control colon segments. Inflammation in the colon was confirmed by measuring body weight, myeloperoxidase activity, colon hypertrophy, mucus staining and inflammatory cells infiltration. These findings demonstrate inflammation‐induced reduction in the expression of CA isoforms which is regulated by TNF‐alpha. Reduction in the binding of CA isoforms with NHE‐1 C‐terminal peptide suggest inflammation‐induced uncoupling of the two transporters. Interaction of CA with the NHE‐1 is important to ensure availability of H+ for NHE‐1 activity. In conclusion both expression and coupling of CA and NHE‐1 are altered and may account for the suppression of NHE‐1 activity in inflamed colon.Support or Funding InformationFinancial support through a grant (MB 03/15) by the Research Sector, Kuwait University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Crystal structure of a dimerization domain of Drosophila Caprin protein
Authors: Zhou Xia , Zhu Jiang , Du Zhihua , Huang Xiaolan 
Published: June 2021

The Drosophila Melanogaster Caprin protein (dCaprin), as well as the human Caprin‐1 and Caprin‐2 proteins, are prototypic members of the caprin (cytoplasmic activation/proliferation‐associated protein) protein family. Caprin proteins contain two highly conserved homologous regions (HR1 and HR2) and C‐terminal RGG motifs characteristic of RNA‐binding proteins. The detailed molecular mechanisms of caprin protein functions remain largely unknown. We had previously determined the crystal structure of a ~120‐residue fragment of Caprin‐1 and Caprin‐2 within the HR1 region. Here, we report the determination of the crystal structure of a corresponding fragment in dCaprin. Our structural studies show that all three structures of dCaprin, Caprin‐1, and Caprin‐2 have a similar novel all α‐helical fold that self‐associates to form a homodimer. Structural comparison reveals that the molecular interactions mediating homodimerization of the three proteins are largely conserved in the three systems. Presence of a dimerization domain within HR1 may represent an evolutionarily conserved feature of the caprin protein family. Interestingly, while the caprin proteins adopt similar overall dimeric structures, the structures have quite different molecular surface properties. The different surface properties of the caprin dimeric structures may dictate their intermolecular interaction with specific protein partners.Support or Funding InformationSIUC Seed GrantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.