bluuebunny/crossref_metadata_2025_split huggingface dataset:
miR144 Regulates Nrf2 Expression In Primary Alveolar Epithelial Cells Derived From HIV‐1 Transgenic Rats
Authors: Fan Xian , Staitieh Bashar B. , Raynor Robert , Kukoyi Abiodun , Guidot David M. 
Published: June 2021

BackgroundDespite anti‐retroviral therapy, individuals living with human immunodeficiency virus (HIV) type 1 infection have impaired anti‐oxidant defenses within their airways and remain at high risk for lung injury. Recently we reported that chronic expression of HIV‐related proteins in HIV‐1 transgenic (HIV‐1 Tg) rats (which models the chronic expression of these proteins in humans living with HIV) inhibits the expression of Nrf2, the master transcription factor that activates anti‐oxidant defenses, in the alveolar epithelial cells and impairs their barrier function thereby leaving the alveolar epithelium vulnerable to injury. However, the mechanism(s) by which HIV‐related proteins inhibit Nrf2 expression have not been identified. MicroRNAs (miRNAs) are post‐transcriptional regulators that have been shown to play critical roles in mediating the redox balance; in particular, miR144 is known to inhibit Nrf2 but a potential role for miR144 in the context of HIV has not been investigated.MethodsPrimary rat alveolar epithelial cells (AECs) were isolated from HIV‐1 Tg rats and wild type (WT) littermates. Total RNA and miRNA were isolated; real‐time RT‐ PCR and western immunoblots were performed to quantify gene and protein expression of Nrf2 and its downstream effectors, as well as key tight junction proteins involved in epithelial barrier function. miRNAs were quantified by qRT‐PCR. In parallel, miR144 expression was increased by treatment with a miR144 mimic or decreased by treatment with a selective miR144 inhibitor (‘antagomir’) to assess the effects of direct manipulation of miR144 expression on Nrf2.ResultsWe first determined that miR144 expression was significantly increased in primary AECs derived from HIV‐1 Tg rats. Further, treating AECs from wild type rats with the miR144 mimic significantly decreased the expression of Nrf2 and its downstream effectors, NAD(P)H dehydrogenase (quinone) 1 (NQO1) and glutamate‐cysteine ligase catalytic subunit (GCLC) and, in parallel, inhibited the expression of the tight junction protein ZO‐1, which is required for alveolar epithelial barrier function. In contrast, treating AECs derived from HIV‐1 Tg rats with the miR144 antagomir significantly increased the expression of Nrf2 and its downstream effectors.ConclusionsHIV‐1 transgene expression increases miR144 expression in the alveolar epithelium, and overexpression of miR144 directly inhibits Nrf2 expression and signaling as well as the expression of tight junction proteins. In contrast, inhibiting miR144 mitigates these pathophysiological effects of HIV‐related proteins on the alveolar epithelium, suggesting that miR144 could be a novel therapeutic target to enhance lung health in individuals living with HIV.Support or Funding InformationThis study is supported by R01 HL 125042.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Environmental Factors Influence α‐synuclein Transport In The Gut‐brain Axis In A Rodent Model Of Parkinsonism
Authors: Bove Cecilia , Anselmi Laura , Travagli R. Alberto 
Published: June 2021

In recent years, idiopathic Parkinson's disease (PD) has been hypothesized to be triggered by the gastrointestinal (GI)‐mediated transport of environmental pathogens; these pathogens then travel retrogradely to the central nervous system in a bottom‐up fashion via the vagus nerve, and induce the typical degeneration of dopaminergic neurons of the substantia nigra pars compacta (SNpc). The presence of misfolded α‐synuclein in SNpc is the characteristic histological hallmark of PD. We have shown previously that oral co‐administration of subthreshold doses of paraquat (1mg/kg) and lectins (0.05%w/v) causes GI dysmotility and may be implicated in PD's etiology.In the present work we aimed to further characterize the effects of this model of Parkinsonism on the gut‐brain axis. Rat groups were gavaged with i) 1% sucrose (control); or ii) 0.5% lectins + paraquat, 1 mg/kg (200μl for 7 days). Motor behavior, and gastric motility/tone were measured 14 and 30 days post‐gavage, via strain gauges sewn to the anterior gastric surface. At the end of the experiment the GI tract and the brainstem were extracted and processed for immunohistochemistry. Organotypic cultures of stomach and small intestine of naïve rats were incubated up to three days in a culture medium containing 0.5% lectins prior to immunohistochemistry. Motor behavioral tests (vibrissae and stepping tests) revealed initial small, albeit significant, motor dysfunctions in the tested rats at 2 weeks post treatment (N=10), the motor dysfunctions were more pronounced at 4 weeks post treatment (N=10). Levodopa (4 mg/kg b.i.d.) re‐established normal motor behavior (N=12).Organotypic cultures revealed that lectins were present in NOS‐ and in NeuN‐ immunoreactive myenteric neurons, as well as in nerve fibers, likely of extrinsic vagal origin. As reported previously (DDW 2017), this experimental model is associated with gastric motor impairment. Tissues from the stomach, small and large intestine of these rats revealed the presence of misfolded α‐synuclein in 40–60 % of myenteric neurons. The presence of α‐synuclein was noted in neurons of the dorsal motor nucleus of the vagus and the SNpc at two weeks post treatment. At four weeks post treatment, the expression of α‐synuclein was more pronounced, and was accompanied by a significant loss of tyrosine‐hydroxylase immunoreactive neurons of the SNpc.The data suggest that prolonged oral exposure to subthreshold doses lectins and paraquat induces a Parkinsonian phenotype that mimics the prodromal GI dysfunctions and α‐synucleopathy progression in the gut‐brain axis observed in patients.Support or Funding InformationNIH DK 55530This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RBM20 Deficiency Postpones Skeletal Muscle Regeneration After Injury and Promotes Fibrotic Tissue Formation
Authors: Rexiati Maimaiti , Sun Mingming , Guo Wei 
Published: June 2021

IntroductionRNA binding motif 20 (RBM20) is a splicing factor with specific expression in striated muscles. It regulates muscle gene splicing and expression. Previous study indicated that RBM20 is associated with myoblast differentiation in C2C12 cell cultures. With higher expression of RBM20, myoblasts were highly differentiated. In this study, we evaluated whether RBM20 regulates muscle differentiation and regeneration in rat muscle injury model.MethodsMuscle injury model was made with 9‐week‐old male wild type (WT) and RBM20 knockout (KO) rats by injecting 0.5ml of 1.2% barium chloride into tibialis anterior and normal saline as control. Tibialis anterior muscles are harvested at 18hours, 3days, 5days, 7days, 14days, and 1month post‐injury (n=5 per time point) and used for western blot, histological and immunofluorescence analyses.ResultsOur results indicated that muscle mass and the regenerating fiber numbers were not significantly different between WT and KO injury during the whole process of regeneration. However, mean cross‐sectional area (CSA) of the regenerating myofibers after 7‐day post injury was significantly smaller in KO when compared to WT, indicating delayed muscle regeneration after injury in RBM20 KO. Further, we found that fibrotic area and TGFβ1 signaling were significantly increased in KO as compared to WT after 14‐day post injury. To test whether RBM20 involves muscle differentiation, we evaluated the expression of Pax7, MyoD, and Myogenin at 5 and 7‐day post injury, we found no changes of these transcription factors at 5 days, but MyoD was significantly reduced at 7 days in KO muscle compared to WT. Interestingly, we found that RBM20 expression was increased with post injury muscle regeneration from 18 hours to 1 month.ConclusionRBM20 is a novel factor that promotes muscle regeneration after injury and protect from fibrotic tissue formation.Support or Funding InformationThis work was supported by the NIFA‐USDA 1009266, National Institutes of Health NIGMSP20GM103432, AHA BGIA and Faculty‐Grant‐in‐Aid from University of Wyoming.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dynamic Assessment of Ca2+ Sensitivity of Isometric Force in Intact Airway Smooth Muscle Using Phase Loop Plots
Authors: Osorio Sara , Han Young , Sieck Gary 
Published: June 2021

A transient elevation of cytosolic Ca2+ concentration ([Ca2+]cyt) triggers a force response in excitation‐contraction coupling. The Ca2+ sensitivity of muscle reflects the relationship between [Ca2+]cyt and force, and in skeletal and cardiac muscle is often defined as the [Ca2+]cyt at which 50% force is generated (ED50). In smooth muscle, Ca2+ sensitivity has been assessed as the ratio of peak force to peak cytosolic [Ca2+]cyt. However, this conventional measurement overlooks the dynamic nature of Ca2+ regulation of force generation. In the present study, in order to provide a dynamic index of Ca2+ sensitivity in airway smooth muscle (ASM), we simultaneously measured [Ca2+]cyt and force and determined their temporal dependence using a phase loop plot. We examined the phase loop plot assessment of Ca2+ sensitivity in ASM under the condition in which the conditional assessment indicated an increase (Calyculin A treatment) in Ca2+ sensitivity. Intact ASM strips were isolated from porcine tracheas obtained from local abattoir. ASM strips were loaded with Fura‐2 AM (Ca2+ fluorescent indicator) and stimulated using 1 mM ACh. Isometric force and [Ca2+]cyt responses were simultaneously measured using a Guth Muscle Research System. ASM strips were exposed to Calyculin A (phosphatase inhibitor; 100 nM for 10 min), which was previously reported to alter static Ca2+ sensitivity. Phase loop plots were calculated from simultaneous [Ca2+]cyt and force responses. The extent of rMLC phosphorylation was determined at different time periods after ACh stimulation using Phos‐tag SDS‐PAGE (SuperSep Phos‐tag; Wako Pure Chemical Industries, Ltd.) and Western blotting. Calyculin A increased ACh‐induced isometric force in ASM strips, but it had no effect on the amplitude of evoked [Ca2+]cyt transients. These results suggested that Calyculin A increased static Ca2+ sensitivity. However, the phase loop plots showed no significant shift in dynamic Ca2+ sensitivity with Calyculin A. These results indicate that the phase loop plot provides a valuable tool to assess dynamic Ca2+ sensitivity of isometric force generation in ASM, which better reflects the complex process of excitation‐contraction coupling in smooth muscle. These results further indicate that dynamic Ca2+ sensitivity of force generation in ASM is independent of rMLC phosphorylation.Support or Funding InformationSupported by NIH grant HL126451 (GCS)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of a Rubric to Evaluate Student Reflection Essays in a Clinical Physiology Course
Authors: Strosdahl Coleman C. , Anderson Lisa Carney 
Published: June 2021

Reflection as a learning technique may be an important strategy for long‐term retention of information. In this study, investigators used a descriptive design to analyze student reflective writings in a two‐course clinical physiology series. In this course series, students completed multiple‐choice exams, first as individuals and second in inter‐disciplinary student groups (biology, physiology, nursing anesthesia and biomedical engineering). Student groups must discuss each question until the group reaches a consensus on the answer. Following the exam students write reflection essays on three exam questions of their choosing. For this assignment, students are encouraged to pick questions that were particularly difficult or that inspired a lot of group discussion. Students were instructed to write about their thought process for choosing their answer and how the group discussion influenced how they thought about the question. The course instructor, also an investigator, collected student writings over a two‐year period, compiled and de‐identified the essays and organized essays according to exam and semester.The course instructor and a student co‐investigator worked as a team to analyze student writings for levels of reflection. First, the investigators adapted a previously published reflection rubric (Hoffman, Shew, Vu, Brokaw & Frankel, 2016) using student writing from the academic year 2015–2016. The investigators analyzed writings separately using the rubric and then met to discuss the essays, reconcile their individual ratings, and revise the rubric as inconsistencies were discovered. In this way, the original rubric was tested and refined. The adapted rubric is a check list of desired essay components: good writing (spelling, grammar, focus), individual reflection, articulation of group process, use of references and insights about future learning. Once a final version of the rubric was agreed upon, the revised reflection rubric was used to evaluate student writings from the academic year 2016–2017 for inter‐rater reliability. There was no correlation between student exam scores and ratings of reflection; this was an unexpected result.Currently the rubric is being used in the 2017–2018 course series to guide student reflection writings.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Morphological characteristics of laboratory rat's primary and secondary lymphoid organs beyond water‐soluble silicon consumption in repeated studies.
Authors: Sergeeva Valentina Efremovna , Gordova Valentina Sergeevna 
Published: June 2021

Recently we have shown that water‐soluble silicon compounds induced certain morphological changes in different lymphoid organs of laboratory rodents. Here we generalized our results received in timeframe from 2011 until 2014.The primary (thymus) and secondary (spleen, Peyer's patches) lymph organs of 6 weeks old white outbred male rats were investigated. The control animals (group 1; n=10) received standardized drinking water ad libitum, the experimental rats (group 2; n=10) got ad libitum the same water containing sodium metasilicate (10 mg/liter silicon content) during 2 months. First study from 2011 was repeated in 2012 and in 2014. All procedures with rats were performed in accordance with the prevailing guidelines for animals care. The thymus, spleen, Peyer's patches were isolated immediately after decapitation, fixed in 10% formalin, and embedded in paraffin. 5μm thick sections were stained with hematoxylin‐eosin and toluidine blue. Photomicrographs from lymph organs were captured with help of light microscope MIKMED 5 under ×1000 magnification and collected for further morphometric evaluation with SigmaScan Pro 5.0 software. The mean areas of the secondary lymphoid nodules were measured in five fields of view with fixed area per animal per group. Standard error of the mean (SEM) were calculated for all means.The comparative morphometric analysis in 2011 revealed that after two month of treatment the germinative center area of splenic and small intestine's secondary lymph follicles in experimental group were 1,2 times larger (p < 0,05), than in control group.The administration of water‐soluble silicon compounds probably forced the physiological involution of thymus by depletion of lymphocyte amount in thymic cytoreticulum. All these data were confirmed in repeated experiments in 2012 and 2014. Moreover, we received the comparable data from thymic mast cell reaction to the silicon consumption in all serial experiments in 2011, 2012 and 2014: ortochromatic mast cells appeared and metachromatic mast cells underwent to degranulation.In 2011, 2012 and 2014, we also noticed, that 20% of experimental animals, which received water‐soluble silicon compounds, had thymic lymph follicles. Known, that such structures appear in the thymus during autoimmune diseases.Our multi‐annual repeated experiments demonstrated that silicon from drinking water probably can influence humoral immunity processes and this fact cannot exclude the role of water‐soluble silicon compounds in the development of autoimmune diseases.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Understanding the development of variable craniofacial phenotypes caused by altered methylation affecting Wnt9b in A‐strain mice
Authors: Green Rebecca M. , Leach Courtney L. , Schmidt Eric , Diewert Virginia , Marcucio Ralph S. , Hallgrimsson Benedikt 
Published: June 2021

Variable clinical presentation is common in many structural birth defects, two children with the same genetic mutation often have different clinical presentation. Therefore, understanding the developmental origin of such variability is of fundamental importance to developing patient targeted treatment, or precision medicine. A/WySn mice are genetically identical, inbred mice, but about 20% of them develop cleft lip and palate. This has been linked to loss of methylation at an IAP retrotransposon near Wnt9b (Wnt9bIAP). Using a newly developed protocol for generating RNA and DNA as well as a 3D microCT scan that can be used to quantify shape, we show that loss of methylation at this locus directly correlates with facial shape in precleft embryos. Line1 methylation is unaffected in embryos with abnormal Wnt9bIAP methylation levels. Wnt9bIAP methylation levels are bimodal, but continuous, as are facial shape, Wnt9b and Axin2 levels. We use all of these factors to build a model of how multiple continuous bimodal inputs lead to a discontinuous phenotype (clefting). We also determine how this model relates to underlying changes in the cell biology in the affected tissue.Support or Funding InformationNIH R01 2R01DE019638 to RSM and BH. NSERC 238992‐17 to BH. Alberta Children's Hospital Research Institute Fellowship to RG.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Tooth Wear Rates for Propithecus diadema in the Fragmented and Pristine Forests of Tsinjoarivo, Madagascar
Authors: Heffernan Katie SusAnn , Samonds Karen , Irwin Mitchell 
Published: June 2021

Primate teeth wear through time in response to their environment. This change in topography has the potential to affect nutrient absorption if mastication is less effective at reducing food particle size. Lemurs exhibit a broad range of diets and life history strategies, as well as adaptations for surviving in an unpredictable environment. Previous studies on diademed sifakas (Propithecus diadema) in Tsinjoarivo, Madagascar show that groups in fragmented forests eat a less diverse diet and use fallback foods year‐round, while sifakas living in pristine forests eat less preferred foods only during the lean season. Groups persist in fragments, yet these dietary changes may increase rates of tooth wear and have consequential deleterious effects on their lifetime fitness and population viability.Previous studies examining tooth wear over time have mainly focused on changes in slope, angularity, and surface relief, but frequently these studies use individuals with estimated age. By examining known‐aged individuals through time, specific patterns of dental wear can be correlated to variation in diet, sex, age, and habitat differences. In this study, dental molds of 75 individuals from 10 groups of Propithecus diadema were taken over a 15‐year period in both the fragmented and pristine forests of Tsinjoarivo. Tooth casts were scanned and analyzed with Geographic Information System (GIS) software to obtain values for relief index, slope, and angularity. All three of these factors decreased with age and show some significance among specific groups of sifakas but results show no effect of overall forest type on rates of tooth wear. Thus, while forest disturbance has demonstrated impact on diet, tooth wear did not accelerate as predicted. Future research should incorporate additional tooth wear data as these known‐aged lemurs advance in years, and examine the functional consequences of tooth wear more directly.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EMBRYO‐FORM PROJECT : STUDY OF THE HUMAN SPINAL CORD VASCULAR SYSTEM
Authors: Barbier Dominique , Renard Yohann , Fauvel Marc , Perez Manuela , Labrousse Marc , Braun Marc 
Published: June 2021

INTRODUCTIONWe proposed to review and illustrate the overall venous human spinal cord vascular embryonic organization using the latest current 3D‐reconstruction possibilities “The Embryo‐form project”MATERIAL & METHODHigh résolution numerised microscopic continuous sections (10 to 20 m) from Carnegie stage XIX to XXIII human embryo were used. Specific image treatment was the following : binarisation‐ thresholding‐labellisation, rigid shifting, median filtration (cubic matrix) (resolution 10mm/pixel) ; DICOM format images are exported for post‐treatment process. Angiographic rendering was obtained thanks to the Eosine/hematoxylin red cell fixation within the vessels.This gathering brought together 20 human embryos prepared in serial histological sections, collected during the mid 50's. We also get access to the Carnegie's embryos collection thanks to her curator Miss E Lockett. 20 Embryos were digitalized with a flatbed scanner at 2400 DPI resolutionRESULTSAt first step, we confirmed the embryonnic status relatively to the Carnegie classification in assessing the cochlear canal orientation. The evolutive morphology of the brain stem primitive network is presented ; at stage XIX, the initial longitudinal veins are separated from the caudal plexus. During the next stages, we demonstrate how they connected to transverse pial anastomoses.The next period (i.e the early and late post‐embryonic period) show these intrinsic pial veins lying between the arteries and the brain surface leading to a progressive simplication pattern. During the stages XX to XXIII, we present the changing morphology of the X, XI and XII nerve ganglion venous plexus. These organisations are compared to the Padget and Streeter works.CONCLUSIONWe present the preliminary work of the cervical spinal cord venous embryogenesis using 3D and angiographic rendering at the early post‐embryonic stagesThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.