bluuebunny/crossref_metadata_2025_split huggingface dataset:
HISTOCHEMICAL ANALYSIS OF STOMACH OF GRASSCUTTER (Thryonomys swinderianus)
Authors: Alawa Judith Nkiruka , Barnabas Kas K , Ibegbu Austin O , Alawa Clement Barikuma 
Published: June 2021

The Grasscutter (Thryonomys swinderianus), is a wild herbivorous rodent peculiar to Africa. It is widely eaten in Nigerian rural communities as a protein source hence its domestication as microlivestock. There is paucity of data on the histochemical studies. This study was aimed at histochemically analyzing the stomach in order to determine if it exhibits regional differences in its qualitative expression of acid, neutral and carboxylated mucin. Seven animals obtained from the wild and domesticated in the animal house of the Department of Human Anatomy, Ahmadu Bello University, Zaria were used. Histologically, stomach of the Grasscutter was simple, glandular and composed of three regions: cardia, fundic‐body and pyloric. Cardia region showed enlongated tubular glands, coiled at the lower end with an opening continuous with the long conspicuous gastric pits. Fundic‐body region showed numerous gastric glands which open into fewer shorter gastric pits while the pyloric region showed simple tubular glands which are highly branched as such obscures the depth of the gastric pits. Histochemically, stomach of the Grasscutter exhibits regional polymorphism in the mucin content with the glands showing weak alcianophilic reactions. Cardia and pyloric regions demonstrated neutral periodic acid Schiff (PAS) reaction characterized by purple coloration thus indicating the presence of neutral mucins. Fundic‐body region exhibited strong PAS positive reaction particularly at the apical cytoplasm of the oxynticopeptic cells indicating the presence of carboxylated mucins. Results demonstrate that regional differences in the expression of mucins occur in the stomach of African Grasscutters, thus this may provide basic information for further nutritional and conservative studies.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Iodine Staining Results in Significant Shrinkage of Sarcomere Lengths in Macaque Biceps Brachii Muscle
Authors: Green Katherine D. , Orsbon Courtney P. , Ross Callum F. , Taylor Andrea B. 
Published: June 2021

Sarcomere lengths are widely used to calculate scaled sarcomere models of whole muscle length‐tension curves. Previous studies have shown that scaled sarcomere models of fresh tissues accurately predict experimental whole muscle length‐tension curves. Diffusible iodine‐based contrast enhanced CT (diceCT) imaging employs iodine staining with Lugol's solution (I2KI) to improve visualization of muscle tissue to the level of individual fascicles, which holds great promise to allow length‐tension modeling of multiple muscles. However, staining skeletal muscles with I2KI causes significant volumetric shrinkage of the muscle, even at levels as low as 2% I2/4% KI, which may affect the validity of length‐tension curves calculated from these tissues. Here we test the hypothesis that iodine staining distorts sarcomere length measurements. We evaluated the impact of I2KI on sarcomere lengths measured from muscle samples of the right biceps brachii from one formalin‐fixed rhesus macaque (Macaca mulatta). We sectioned the muscle into a primary control (no I2KI) and two experimental groups: one treated with 20% (w/v) I2KI and one pretreated with a sucrose prewash (20% w/v) prior to iodine staining. The muscle sections then were chemically digested in 70% nitric acid in saline and mounted on slides. A minimum of three sarcomere length measurements was taken per fiber using laser diffraction and the averages used for ANOVA and post hoc t‐tests. The results show that iodine staining significantly shrinks sarcomere lengths (2.08 ± 0.22 μm) compared with primary controls (2.45 ± 0.25 μm) (p < 0.001). The sucrose prewash group also experienced significant shrinkage (2.13 ± 0.23 μm). These data demonstrate the need for methods which correct for this pronounced sarcomere shrinkage before iodine stained skeletal muscle samples can be used to calculate whole muscle length‐tension curves.Support or Funding InformationNational Science Foundation Grant, Division of Behavioral and Cognitive Science, DDRI Bio Anthro, Award #1732175This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mesterolone Increases Satellite Cell Number without Inducing Hypertrophy in Type I Oxidative Muscle Fibers
Authors: Allouh Mohd (Mohammed) Zohair 
Published: June 2021

Mesterolone is a synthetic oral anabolic androgenic steroid used to treat hypogonadism. Limited information is available about how this drug exerts its effects on skeletal muscle. Satellite cells (SCs) are mononuclear myogenic stem cells that contribute to postnatal skeletal muscle growth and maintenance. As SCs' activation and subsequent differentiation to new myonuclei is a major event during skeletal muscle hypertrophy, this study investigated the influence of Mesterolone on SC distribution within type I oxidative fibers of the adult avian skeletal muscle. Immunocytochemical techniques and morphometric analyses were used to calculate the fiber size, and numbers of SCs and myonuclei within the anterior latissimus dorsi (ALD) muscle of the chicken. The ALD is a slow (tonic) contracting muscle that is made up exclusively of slow‐twitch type I oxidative red fibers. The number of SCs was significantly (P<0.01) increased by very high doses (12 and 16 mg/kg) of Mesterolone administration. Similarly, the number of myonuclei was increased by administering the high doses of Mesterolone. However, this increase did not reach significant levels except at the Mesterolone dose of 12 mg/kg as the number of myonuclei increased significantly (P<0.05) by 32% compared to the number of myonuclei in the control birds. In addition, no significant (P>0.05) differences were detected in the fiber area and the myonuclear domain size between control and Mesterolone‐treated birds. This study indicates that Mesterolone treatment can increase the numbers of SCs and myonuclei without inducing hypertrophy in type I fibers of the adult avian skeletal muscle. This is because SCs in type I fibers are, most probably, required for fiber maintenance rather than hypertrophy since these fibers have a higher rate of protein breakdown because they are the ones that mostly used in daily routine activities.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Intrinsic Cardiac Ganglionic Neurons Projecting to the SA node in the Rat and Pig Hearts: Retrograde Labeling and Neurolucida 3‐D Reconstruction
Authors: Chen Jin , Hanna Peter , Ardell Jeffrey L , Hoover Donald B , Vadigepalli Rajanikanth , Schwaber James , Shivkumar Kalyanam , Cheng Zixi J 
Published: June 2021

Intrinsic cardiac ganglionic neurons (ICNs) within the heart are essential for regulation of cardiac function. ICNs include afferent, efferent neurons and interneurons and constitute a local integration center. In rat and pig hearts, there are multiple cardiac ganglia. Among these ganglia, the largest ganglionated plexus in the rat heart and the right atrial ganglionic plexus (RAGP) in the pig heart are near the sinoatrial node (SAN). However, it is not clear whether both plexi project to the SAN. In this study, retrograde tracer Fast DiI was injected into the SAN in rats (n=5) and a pig. SAN cells were identified by immunostaining for pacemaker channel HCN4. Fluorogold (FG) was injected (i.p.) to label all of the ICNs in rats. FG‐ and DiI‐labeled ICNs were identified and analyzed using a confocal microscopic system and a Neurolucida Tracing and Digitization System. Sixteen days after Fast DiI injection in rats, the animals were perfused with paraformaldehyde, and the whole‐mount atria were prepared. There were multiple ganglionated plexi and the DiI‐labeled neurons were mainly found in the largest ganglionated plexus near the SAN in all these rats. Twenty‐one days after Fast DiI injection in the pig, the animal was sacrificed, and the RAGP was removed and sectioned at 40 μm. Numerous DiI‐labeled neurons were found in the RAGP of this pig. Interestingly, these DiI‐labeled neurons were organized in five spatially‐separated columns perpendicular to the epicardial surface. Our data suggest that the largest ganglionated plexus in rat indeed projects mainly to the SAN, and the pig RAGP projects very heavily to the SAN. The functional significance of this large ganglionated plexus in rat and the five spatially‐separated columns of RAGP ICN projecting neurons in the pig will be elucidated in the future.Support or Funding InformationSupported by NIH SPARC grant OT2OD023848, NIH R15HL137143‐01A1.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Periocular Mesenchyme (POM) Contributes to Anterior Eye Structures
Authors: Svoboda Kathy K H , Thomson Robert , Serrano Maria Juliana , Zhao Hu , Petroll Matthew 
Published: June 2021

PurposeEstablish a new glaucoma animal model. The objective of this project is to determine if Gli1 positive cells contribute to the POM and anterior eye structures by using inducible Gli1‐CreERT2; tdTomatoflox (Gli1‐tdTomato) mouse model to serve as a probe for anterior eye development. Pitx2, FOXC1, and FOXC2 are expressed in the periocular mesenchyme, and are critical for anterior eye development. Mutations in these genes are associated with a broad spectrum of abnormalities in these tissues. A second objective is to establish that FOXC1, FOXC2, and Pitx2 are expressed in the Gli1 positive cells.MethodsGli1‐tdTomato expressing mice were induced at different stages with tamoxifen. Eyes were examined that were induced on embryonic (E13.5) and postnatal days: P3, P7, P30 and sacrificed several days to weeks later. Mouse tissues were fixed and prepared for frozen or paraffin sectioning. Tissue antigen retrieval was done using citric acid buffer. Indirect immunohistochemistry for FOXC1, FOXC2, and Pitx2 was completed. Some samples were counterstained with phalloidin. Sections were analyzed with confocal microscopy for spatial distribution of the Gli1+ labeled cells and FOXC1, FOXC2, and Pitx2.ResultsIn Gli1‐tdTomato mice induced on P3, P7, and P30, the POM and anterior angle of the eye, eyelids and optic nerve were labeled several days to 3 weeks later. In contrast, the cornea, lens and sclera did not have positive cells for Gli1. Expression levels for FOXC1, FOXC2, and Pitx2 were generally higher in mice harvested at younger postnatal ages. After postnatal day 30, the expression decreased with little to no expression of FOXC1 and FOXC2 while Pitx2 expression remained relatively constant in the anterior angle tissues and was expressed in some Gli1 positive cells.ConclusionThis inducible Gli1‐tdTomato mouse model may be an excellent system for determining the role of POM in anterior eye development. The elegance of this model is it can be activated at specific developmental stages and Gli1+ cell derivatives can be tracked and other detection probes (phalloidin, nuclear stains, Immunohistochemistry etc), can be incorporated to determine cell characteristics or over‐all tissue structure. Our results provide evidence that POM cells are contributing to the development of the anterior angle chamber that forms between P3‐P10 in mice.Support or Funding InformationBiomedical Sciences Department Seed Grant (K. Svoboda), NIH K08DE025090 (H. Zhao)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Emergent Properties of Facial Morphogenesis Regulated by Fgf Signaling
Authors: Marcucio Ralph , Hu Diane , Hallgrimsson Benedikt , Young Nathan 
Published: June 2021

Craniosynostosis is a severe disorder that may be caused by activating mutations in Fibroblast growth factor receptors (FgfRs) that affects development of the skull and is characterized by premature fusion of the cranial sutures. In addition to dysmorphology of the skull, affected individuals also have facial dysmorphology. We have previously used an avian model to explore the role that a disease‐causing allele of FgfR2 (FgfR2C278F) plays in producing malformations of the middle and upper face at early stages of development. In that work we observed a wider midface, which resembles what is observed in human patients, and this was directly related to decreased cell proliferation and a disrupted net polarization of the mesenchymal cells in the growing facial primordia. We have extended this work by examining the role of the Map kinase, PLC‐gamma, and PI3K pathways downstream of Fgf signaling in producing facial dysmorphology, as well as directly blocking FgfR activation. We implanted beads soaked in small molecular inhibitors (MEK1/2: U0126, PLC‐gamma: U‐73122, PI3K: LY 294002, FgfR activation: SU5402) into the right side of the developing avian face at HH22 and examined morphological and cellular outcomes at various times of development. First, we determined that each inhibitor substantially and specifically down‐regulated its respective pathway using immunohistochemistry to assess expression of down‐stream targets of activation of each pathway. We then determined that each pathway significantly reduced cell proliferation and altered cell polarization in the mesenchyme. Blocking each pathway also created severe craniofacial malformations that were observed at early and late time points. There are many inputs to each of these pathways, and our data suggest that each participate in regulation of a similar set of cellular processes that contribute to the emergence of morphogenetic processes in facial development.Support or Funding InformationNIH: R01DE019638, R01DE018234, R21DE028198This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of alpha‐amylase during murine palatine gland development: A light microscopic immunocytochemical study
Authors: Kalfut Mahasin , Alhazmi Ahlam , Alhakami Khalid , Hakami Zaki 
Published: June 2021

IntroductionSalivary alpha‐amylase is one of the major enzymes in the oral cavity. Beyond its primary function of initiation of digestion in the oral cavity, it is involved in defense against bacteria. Alpha‐amylase, a product of serous cells, is the most abundant protein of parotid saliva and it is also secreted by sublingual glands. However, its presence in the minor glands, including palatine glands, has been contradictory. Therefore, this study was conducted to observe the expression of alpha‐amylase at different developmental stages of rat palatine glands using light microscopic immunohistochemistry.Materials and MethodsAn avidin‐biotin technique was used to detect the binding of alpha‐amylase on sagittal frozen sections of palatine glands of pre‐ and post‐natal rats.ResultsPositive reaction of alpha‐amylase was observed at the basolateral and perinuclear membranes of the developing acinar cells at the embryonic day 20 (E20). At birth, in addition to the basolateral membranes, weak cytoplasmic reaction labelling a few mucous cells was observed. Amylase reactivity increased with maturation and reached peak reactivity at postnatal day (PN) 10 and 14, thereafter, decreased into adulthood. Serous cells expressed amylase starting from PN7. Whereas ducts showed variable reaction on the luminal membrane starting from PN3. In adult rats, heterogeneous distribution was revealed between the anterior and posterior portions of palatine glands, as amylase showed more reactivity in the posterior portion.ConclusionThe expression and distribution of alpha‐amylase showed significant modifications during cytodifferentiation and maturation of the palatine glands.Support or Funding InformationNoneThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cold‐Inducible RNA Binding Protein (CIRP) Decreases Inflammation and Breast Tumor Growth
Authors: Lujan Daniel Albino , Hathaway Helen J , Kusewitt Donna F , Hartley Rebecca S 
Published: June 2021

RNA binding proteins (RBPs) post‐transcriptionally regulate gene expression by associating with the regulatory sequences in the untranslated regions of mRNAs. RBPs are often disrupted in cancer, but current knowledge of their role in regulating cancer‐associated genes is largely based on in vitro studies. Cold‐inducible RBP (CIRP) is a stress‐induced RBP that was recently shown to modulate inflammation in response to cellular stress. CIRP expression is altered in several cancers, including breast cancer. The effect of CIRP on inflammation in breast cancer is not known. This study tested the hypothesis, based on our preliminary data and published studies that CIRP changes the inflammatory profile of breast tumors from pro‐ to anti‐tumorigenic, and that CIRP expression and this altered inflammatory milieu are associated with lower tumorgenicity. To test this hypothesis a transgenic mouse overexpressing CIRP in the mammary epithelium was crossed with the PyMT mouse model of breast cancer. Effects on both early (7‐week‐old mice) and late (14‐week‐old mice) tumorigenesis and inflammation were assessed in CIRP/PyMT and PyMT mice. Mammary glands from 7‐week‐old CIRP/PyMT mice had 50% fewer mammary intraepithelial neoplasias than PyMT mice while 14 week‐old CIRP/PyMT mice had slower tumor growth kinetics, smaller tumors, decreased tumor burden and decreased pulmonary metastasis compared to PyMT mice. Importantly, CIRP protein level correlated with decreased tumorigenesis in CIRP/PyMT mice. To assess effects on the inflammatory environment, mammary glands from 7 and 14‐week‐old mice were incubated in medium overnight and Luminex cytokine assays were performed on the medium. Luminex assays were also performed on sera from 14‐wk mice to assess effects on systemic inflammation. There was a decrease in cytokines that promote inflammation, angiogenesis and metastasis at both timepoints for mammary glands, and in 14‐week sera. Overall, these data support a role for CIRP as a potent anti‐tumor molecule that suppresses local and systemic pro‐tumorigenic inflammation. To begin to probe whether CIRP's effects on cytokine levels were cell‐autonomous, we performed in silico analysis of CIRPs binding affinity for individual targets using the web‐based beRBP tool (Yu et al.). CIRP was not predicted to bind the cytokine mRNAs so may be acting in a non‐cell autonomous fashion.Support or Funding InformationNational Institutes of Health Grant/Award Number: F31‐CA213933American Association of Anatomists, Fellows Grant Award ProgramThis 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 a GDF5 Associated Enhancer During Limb Development
Authors: Cooper Allen M. , Shankel Matthew , Haro Endika , Pira Charmaine U. , Oberg Kerby C. 
Published: June 2021

Growth differentiation factor 5 (GDF5) is an anabolic growth factor that plays a critical role in the formation, maintenance and repair of bones and joints. Reduced levels of GDF5 are associated with accelerated degradation of joint cartilage, a disease called osteoarthritis (OA). Identification of factors involved in GDF5 regulation may offer therapeutic targets for treating or preventing OA. A GDF5 Associated Regulatory Region (GARR) has been previously identified as a 900 bp enhancer located 78 kb downstream of the GDF5 locus. GARR contains a number of potential transcription factor binding sites, including sites for odd‐skipped related (Osr) zinc‐finger and Sry box (Sox) transcription factors. As Osr1/2 and Sox11 are important for normal Gdf5 expression, and their expression patterns overlap that of Gdf5, we hypothesized that mutation of OSR and SOX binding sites within GARR would disrupt enhancer activity.To identify whether putative OSR and SOX transcription factor binding sites contribute to GARR activity, we performed site‐directed mutagenesis on both potential OSR binding sites (GARRmutOSR), or on both potential SOX binding sites (GARRmutSOX) in the GARR‐tk‐EGFP reporter construct (GARRcontrol). We used targeted regional electroporation (TREP) to introduce control and mutated reporter constructs into Hamburger‐Hamilton stage 23 chicken limb buds near the developing elbow joint. Transfection efficiency was determined by co‐transfection with a β‐actin promoter‐driven RFP construct. We determined GARR activity by fluorescence microscopy, following 48 hours of incubation after TREP.Comparison of GARRcontrol and GARRmutOSR demonstrated similar GFP fluorescence intensity localized to the elbow joint. In contrast, fluorescence of GARRmutSOX was markedly reduced relative to GARRcontrol, although similarly restricted to the elbow. Though Osr transcription factors are associated with Gdf5 expression, mutation of the OSR binding sites in GARR was insufficient to substantially alter the level of GARR activity. Mutation of SOX binding sites, however, dramatically reduced GARR activity, implicating SOX transcription factors as critical regulators of GDF5 expression.Support or Funding InformationThe research was funded in part by a grant from the LLU Pathology Research Endowment.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Circularization of linear chromosomes and telomerase RNA gain‐of‐function mutations in Saccharomyces cerevisiae
Authors: Mefford Melissa Anne 
Published: June 2021

Telomeres are repeated DNA sequences located at the ends of linear eukaryotic chromosomes. While they act as protective caps, they cannot be fully copied by the DNA replication machinery. To overcome this end‐replication problem, most eukaryotic organisms express the ribonucleoprotein enzyme complex telomerase. I will present preliminary results investigating basic, broad questions of telomere and telomerase evolution in the budding yeast, Saccharomyces cerevisiae. First, we are genetically engineering strains with circularized versions of each of the 16 chromosomes to assess the viability and potential fitness effects on the organism. Second, we are screening for gain‐of‐function mutations in telomerase RNA to gain further insights into how this component contributes to enzyme function.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.