bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gap junction blockade in the locus coeruleus (LC) decreases the hypercapnic ventilatory response
Authors: Patrone Luis Gustavo A , Bicego Kenia C , Hartzler Lynn , Putnam Robert W , Gargaglioni Luciane H. 
Published: June 2021

The LC has an important role in central chemoreception and its contribution decreases with development. In neurons from young rats (P9 or younger), the % of LC neurons that increase firing rate in response to hypercapnia is 85% while it is only about 45% in neurons from rats P10 or older. Carbenoxolone (CARB ‐ gap junction blocker) has very little effect on this response in neurons from the youngest rats, with the % responding dropping only to 70% in the presence of CARB. However, CARB decreases the % responding by half in older animals, dropping it to 21%. We evaluated the participation of gap junctions in the CO2 ventilatory response in conscious adult rats by bilaterally microinjecting CARB (1mM or 3mM / 0.1 μL) or vehicle (artificial cerebrospinal fluid) into the LC of Wistar rats. Bilateral gap junction blockade in LC neurons did not affect resting ventilation. However, the increase in ventilation produced by hypercapnia (7% CO2) was reduced by 20% after CARB injection (1904.5 ± 95.2 in vehicle group vs 1518.0 ± 76.1 mL/kg/min in CARB 3mM group and 1468.3 ± 122.2 mL/kg/min in CARB 1mM) due largely to a decrease in respiratory frequency. The results suggest that gap junctions modulate the hypercapnic ventilatory response, but that they are not involved in respiratory control during normocapnia, in the LC of rats older than P10.Financial support: INCT‐Fisiologia Comparada, FAPESP, CNPq, and NIH Grant HL56683.
Functional confocal live cell imaging of the pulmonary neuroepithelial body microenvironment in GAD67‐GFP mice
Authors: Schnorbusch Kathy , Brouns Inge , Lembrechts Robrecht , Pintelon Isabel , Timmermans Jean-Pierre , Adriaensen Dirk 
Published: June 2021

As integrated components of the airway epithelium, pulmonary neuroepithelial bodies (NEBs), composed of clusters of neuroendocrine cells, are ideally located to sense changes in the airway environment. Connections of NEBs with different populations of mainly vagal sensory nerve terminals allow fast transduction of information to the CNS. NEBs are dispersed in the airway epithelium and inaccessible in vivo. The selective identification of NEBs is a prerequisite to enable functional molecular live cell imaging in our ex vivo lung slice model, but the vital dye 4‐Di‐2‐ASP that we used so far has been criticized. In this study the lung slice model was further strengthened using knock‐in mice that express GFP driven by the promoter of glutamate decarboxylase 67 (GAD67), the rate‐limiting enzyme of GABA synthesis. Highly selective expression of GFP in NEB cells in mouse lungs allows unequivocal selection. Since ex vivo vibratome slices of GAD67‐GFP mouse lungs could be loaded with red‐fluorescent functional probes, and proof‐of‐concept experiments were successful, this model will certainly boost future functional studies of NEBs in health and disease. Simultaneously the model enables to explore the significance of a GABAergic signaling system in the NEB microenvironment.Support: IWT fellowship SB/81162 (RL); FWO grants G.0081.08 (DA) and G.0589.11N (DA, JPT); UA grants GOA BOF 2007 (DA) and KP BOF 2006 (IB)
Cerebrovascular Consequences of Obstructive Sleep Apnea
Authors: Crossland Randy F. , Durgan David J. , Lloyd Eric E. , Phillips Sharon C. , Marrelli Sean P. , Bryan Robert M. 
Published: June 2021

Obstructive sleep apnea (OSA) is an independent risk factor for stroke. However, little is known about how OSA affects the cerebral circulation. The goal of this project was to test the following hypothesis: Chronic exposure to apnea produces hypertension and alters nitric oxide (NO) mediated dilation in cerebral arteries. For these studies, we developed a novel model of OSA in rats. Airway obstruction was induced by a chronically implanted, inflatable endotracheal obstruction device. We induced 30 apneas (8 sec duration each)/hr during 8 hrs of the sleep cycle in free‐ranging rats. Apnea caused significant alterations in blood gasses [baseline vs. end apnea‐ pO2 (mmHg): 127 ± 2 vs. 67 ± 2; pCO2 (mmHg): 43 ± 1 vs. 51 ± 1; and pH: 7.46 ± 0.002 vs. 7.38 ± 0.01; n=5, p≤0.003]. Apnea decreased hemoglobin O2 saturation by 12 ± 1% (n=5, p=0.001 vs. Sham) and produced exaggerated negative airway pressures (−16 ± 0.7 mmHg; n=7, p<0.001 vs. Sham). Two weeks of apnea produced a 24‐hr mean blood pressure (BP) pattern similar to non‐dipping hypertensives (i.e. ‐ <10% BP dip during the sleep phase). After one month of apnea, NO‐mediated dilations to ATP in isolated cerebral arteries were attenuated by 20 ± 2% (n=4, p<0.02 vs. Sham). Using a novel model of OSA, which recapitulates OSA in humans, we demonstrate that endothelial dysfunction occurs by one month, likely increasing the vulnerability of the brain during stroke.Funded by NIH R21 HL102778
Oxidative stress in hypothalamic neurons may explain the impaired vasopressin secretion observed during sepsis
Authors: Basso Paulo José , Oliveira-Pelegrin Gabriela R. , Rocha Maria José A. 
Published: June 2021

Both clinical and experimental studies denoted an impaired vasopressin (VP) secretion in the late phase of sepsis, despite progressive hypotension. One hypothesis to explain this is an excessive production of nitric oxide in the brain, which may induce oxidative stress and apoptosis in vasopressinergic neurons. Our aim was to investigate the occurrence of mitochondrial oxidative stress and apoptosis in magnocellular neurons of the supraoptic nucleus of septic rats.Male Wistar rats (250±30g) were anesthetized and perfused 24 or 48 hours after cecal ligation puncture (CLP) to induce sepsis, or following sham operation. Brains were removed and processed for immunodetection of hypoxia‐induced factor 1 subunit α (HIF‐1α), Bax, cytochrome‐c (cyt‐c) and cleaved caspase‐3 (casp3), or TUNEL assay.A granular or punctate staining for cyt‐c and a homogeneous one for Bax was observed in sham rats. These patterns were inversed in some of the septic rats indicating translocation in opposite directions for both proteins with respect to the mitochondria. In these same animals, immunofluorescence for casp‐3 and HIF‐1α reached higher levels compared to sham rats. Additionally, a positive TUNEL staining was observed.We conclude that sepsis can induce oxidative stress causing apoptosis in hypothalamic magnocellular neurons, which could explain the basal VP levels denoted in the late phase.Financial support: FAPESP, CAPES.
Morphological Changes of the Membranous Labyrinth in Meniereˈs Disease
Authors: Tanioka Hisaya , Kaga Kimitaka 
Published: June 2021

ObjectivesTo image morphological changes of the membranous labyrinth in Meniere's disease using direct volume rendering by temporal bone high‐resolution CT exam. 3D reconstructed images of endolymphatic hydrops with direct volume rendering generate morphological changes in Meniere's disease.Study DesignIn vivo retrospective clinical study during pre‐attack, attack and recovery periods in Meniere's disease.PatientsPatients with Meniere's disease who were examined at three stages, namely, pre‐attack, attack and recovery period.MethodsAll CT images were acquired on a row CT scanner. The axial data were applied to create 3D volume rendering images on the CT work station for post processing with the factory preset soft ware.Results3D images of the membranous labyrinth in Meniere's disease demonstrate different morphologic change in proportion to periods. On attack period, the saccule and utricle was enlargement in comparison with pre‐attack period and recovery period. Patients were treated by increases in vestibular, systematic blood flow and decreases in blood pressure.ConclusionWe reported the first visualization of endlymphatic hydrops in patients with Meniere's disease that were consistent with periods.
Individual craniofacial bone volumes and relative densities in postnatal mice provide valuable phenotypic information
Authors: Percival Christopher , Wang Yingli , Zhou Xueyan , Jabs Ethylin Wang , Richtsmeier Joan T 
Published: June 2021

The objective of this study was to develop an efficient method, using Avizo software (VSG), to measure the volume and relative bone mineral density of individual craniofacial bones in early postnatal mice from micro‐computed tomography images. Manually labeled individual bones from representative specimens were expanded to define the theoretical space that a bone would occupy in a standard orientation. These models were transformed into the space of related target specimens and used to identify all voxels above three phantom based on minimum bone density thresholds for each bone of the target specimen. Total bone volumes, medium/high density bone volumes, and ratios of these volumes (relative BMD) were produced for Fgfr2Y394C/+ Beare‐Stevenson cutis gyrata syndrome model mice and unaffected littermates at postnatal day 0 and 8. This provided phenotypic and developmental information unavailable from landmark based morphometrics that can help target regions and developmental times for further analyses. This procedure requires <5% of the time required for manual segmentation. Error levels are acceptable for summary analysis of bone volume and relative density, with higher error for older specimens, smaller bones, and bones surrounding teeth. This work is supported in part by a grant from the National Institute of Dental and Craniofacial Research to JTR (R01‐DE018500, ‐DE18500‐S1, ‐DE18500‐S2).Grant Funding Source: National Institute of Dental and Craniofacial Research
Esrp1 and Esrp2 expression in the developing midface and kidney of mice with frontonasal dysplasia and renal hypoplasia
Authors: Hong Trudy , Hynd Thomas , Lozanoff Scott 
Published: June 2021

The Brachyrrhine (Br) mutant mouse displays heritable frontonasal dysplasia (FND) and renal hypoplasia (RH). Previous DNA microarray analysis on embryonic Br facial explants showed a misexpression of several genes, two of which are Esrp1 and Esrp2. Together, the Esrp genes play an essential role in the regulation of epithelial‐mesenchymal transition—an important process in craniofacial and kidney development. The primary objective of this study is to determine if Esrp1 and ‐2 contributes to a developmental pathway leading to FND or RH in the Br mouse and, specifically, if there is a difference in Esrp1 and ‐2 expression in the developing midface and kidneys of wild type (WT) and Br adults and embryos. WT and Br/Br E11.5 medial nasal prominences (MNP) and E15.5 and E18.5 kidneys were dissected and RNA extracted for qPCR expression analysis. Preliminary results show minimal difference in Esrp1 and ‐2 expression in MNPs of WT and Br embryos, and an increase in Esrp2 expression from E15.5 to E18.5 WT kidneys. Future work involving qPCR and immunohistochemistry will be completed to better understand the expression patterns of these two genes. Results from this study will contribute to our understanding of the genetic basis of craniofacial and kidney morphogenesis.Grant Funding Source: 1R01‐DK‐064752 (SL)
FGF signaling specifies pre‐chondrogenic identity in neural crest derived mesenchyme, but initiation of chondrogenesis requires BMP4 signaling
Authors: Kumar Megha , Ray Poulomi , Chapman Susan C. 
Published: June 2021

The chick middle ear bone, columella, derives from neural crest mesenchyme located proximally in the second pharyngeal arch. Subjacent pharyngeal endoderm is hypothesized as the source of the local signals that specify the pre‐chondrogenic identity of this tissue. Sox9 is induced and maintained in pre‐chondrogenic cells during condensation formation and endochondral ossification. We show that pharyngeal endoderm was sufficient, but not necessary for specifying pre‐chondrogenic identity. Many Fgf genes are expressed specifically in the pharyngeal endoderm subjacent to neural crest derived mesenchyme. Our results show that FGF signaling is both sufficient and required for specification of Sox9 expression and specification of pre‐chondrogenic identity. Further, FGF signaling, by itself, is insufficient for the induction of the chondrogenic marker, Col2a1. BMP4 signaling can induce Sox9 expression, but only in combination with FGF signaling induced Col2a1 expression and, thus, chondrogenesis. Given the spatio‐temporal expression patterns of FGFs and BMPs in the pharyngeal arches, we suggest that this may represent a general mechanism of specifying pre‐chondrogenic identity and chondrogenesis.Grant Funding Source: NIH/NIDCD
Mesenchymal condensation sets the stage for intramembranous bone development
Authors: Jabalee James , Franz-Odendaal Tamara 
Published: June 2021

Mesenchymal condensation is one of the earliest, and most critical, stages in the formation of intramembranous (directly ossifying) bone. It is during the condensation phase that key skeletogenic genes are first upregulated and during which collagen I, the scaffold on which the inorganic portion of bone will be deposited, is first established. Thus, morphogenesis and tissue specific gene expression occur concurrently. Previously, we used microarray analysis and in situ hybridisation to determine which genes are important and at what stage of development these genes act during intramembranous ossification in chick and zebrafish. In the current study, we utilize a combination of light and electron microscopy to better understand the patterning and polarization of cells in early skeletogenic condensations. We compare neural crest derived intramembranous bones, specifically the scleral ossicles of the chick (Gallus gallus) and the opercula of the zebrafish (Danio rerio). While our preliminary results in the chick suggest osteoblasts are polarized toward the center of the early condensations, more data is required to make a solid conclusion. Data collection in zebrafish is also ongoing. Combined, these studies provide a means to correlate gene expression and morphogenesis during intramembranous bone formation.Grant Funding Source : Nova Scotia Health Research Foundation (Canada) ; Natural Sciences and Engineering Research Council of Canada
Embryonic transcription factor upregulation during normal lactation and breast oncogenesis
Authors: Hassiotou Foteini , Beltran Adriana , Trengove Naomi , Lai Ching Tat , Hartmann Peter , Filgueira Luis , Blancafort Pilar 
Published: June 2021

In addition to their physiological role during pregnancy/lactation, mammary stem cells (MaSCs) are primary targets of malignant transformation, giving rise to invasive breast carcinomas. To pinpoint the molecular switches that transform these cells, we must understand the basic machinery that controls their self‐renewal. We non‐invasively accessed lactation‐associated MaSCs via breastmilk. A cell population was identified expressing the pluripotency transcription factors (TFs) Oct4, Sox2 and Nanog, which generated monolayers and spheroids in 2D and 3D culture, respectively, expressing these TFs. Culture in differentiation conditions resulted in differentiation into cells originating from the three germ layers. Examination of normal resting and lactating human breast tissues revealed that these TFs are upregulated during lactation. Lentivirally‐transduced Oct4‐overexpressing mammary cells from the resting gland bypassed cellular senescence and generated subcutaneous tumors with colonization capabilities when injected in nude mice. Examination of different types of breast tumors showed expression of these TFs only in invasive basal tumors with lactating features. It is concluded that these embryonic TFs play important regulatory roles in the normal expansion of the breast during pregnancy/lactation, but also act as oncogenic targets in certain types of breast cancers.Grant Funding Source : Women and Infants Research Foundation (FH); Medela AG (PH); 1R01CA125273 and 3R01CA125273‐03S1 and Department of Defence grant W81XWH‐10‐1‐0265 (PB)
Cell fusion phenomena was detected after xenogenic intra‐uterus transplantation of Ds‐red porcine amniotic fluid derived stem cells into EGFP bearing mice
Authors: peng shao-yu , Chou Chih-Jen , Wu Shinn-Chih 
Published: June 2021

Amniotic fluid derived stem cells (AFSCs) are derived from amniotic fluid of developing fetus and can be given rise to three germ layers ‐ endoderm, mesoderm and ectoderm. Intra‐uterus transplantation (IUT) is an approach to cure the inherited genetic defect for the fetus during the gestation period of the pregnant female. Certain diseases such as osteogenesis imperfect, muscular destrophy, ect. were successfully treated with stem cells to ameliorate the syndrome. The purpose of this study was to investigate the bio‐distribution and engraftment of porcine amniotic fluid derived stem cell (pAFSCs) after IUT and further determine the fate of the donor cells through Ds‐red harboring pAFSCs inject into EGFP‐harboring fetus of the pregnant mice. The 12.5 days pregnant mice were performed the IUT surgery by pAFSCs injection. One month after birth, the mice were sacrificed. Several portions from different organs were sampled for histological examination. Experiment results showed that the migration of Ds‐red pAFSCs were expressed most significantly in the intestines. Furthermore, the EGFP signal and RFP signal were co‐expressed in the detected cells. It is concluded that the exogenous implantation of pAFSC was fused with recipient cells instead of differentiation or maintain the undifferentiated status retaining in the tissue.Grant Funding Source : National Taiwan University
Using neural stem cells as an anti‐cancer drug delivery vehicle for treating human breast cancer
Authors: Reuter Ashley , Davis Kelsey , Rachakatla Raja Shekar 
Published: June 2021

The broad, long‐term objective of this lab was to use C 17.2 murine neural stem cells (NSC) as a targeted, drug delivery vehicle to treat tumors. Our goal was to determine if NSC can be used as a drug delivery vehicle to administer interferon beta (IFN‐β) directly to MDA 231 human breast cancer and to look at the therapeutic ability of NSC transduced with IFN‐β (NSC‐IFN‐β) on MDA 231 cells. We addressed two specific aims. Specific aim 1 determined if NSC migrate towards MDA 231 cells in vitro. A transwell plate was seeded with selected concentrations of MDA 231 cells in the lower chambers and NSC in the upper chambers. After 24 hours, the migrated NSC were counted using confocal microscopy.ResultsThere was a significant increase (P<0.001) of NSC migration towards 10,000 and 100,000 MDA 231 cells compared to control. Specific aim 2 determined if NSC‐IFN‐β migrate towards and kill MDA 231 cancer cells in vitro. A transwell plate was seeded with MDA 231 cells in the lower chambers and NSC (control) or NSC‐IFN‐β in the upper chambers. After 48 hours, the live and dead cells were counted.ResultsThe number of live cells significantly decreased (P<0.001) when using NSC‐IFN‐β compared to control.ConclusionNSC and NSC‐IFN‐β migrate towards MDA 231 human breast cancer and NSC‐IFN‐β significantly reduced the number of cancer cells. Future studies include testing NSC‐IFN‐β efficacy on human breast cancer in vivo.Grant Funding Source : A. T. Still University Graduate Program
Effects of Mesterolone on satellite cell distribution and myonuclear number in maturing skeletal muscle fibers
Authors: Allouh Mohammed Zohair , Aldirawi Mohammed Hasan 
Published: June 2021

Mesterolone is a synthetic oral anabolic steroid usually used to treat hypogonadism. Mesterolone abuse has been frequently reported in human and equine sports, in which it is used to increase muscle mass and strength. However, little is known about the effects of mesterolone on satellite cells (SCs), which permit muscle growth by forming new myonuclei. This study tests the hypothesis that mesterolone induced muscle hypertrophy is associated with an increase in the number of SCs. Using immunocytochemical techniques and morphometric analyses, fiber size and numbers of SCs and myonuclei in the pectoralis muscle of maturing chickens were quantified. Mesterolone significantly increased fiber size by approximately 36% (P<0.01). In addition, numbers of SCs and myonuclei were significantly increased by mesterolone administration (P<0.01). These results demonstrate that mesterolone‐induced skeletal muscle hypertrophy is associated with increased numbers of SCs during maturation. Funds for this study were provided by a grant from the Deanship of Research at Jordan University of Science & Technology (25/2010).Grant Funding Source: Deanship of Research, JUST
Acute Inhibition of Connexin43 Expression Promotes Vesicant Wound Repair in a Nitrogen Mustard Induced Hairless Mouse Cutaneous Injury Model
Authors: Chang Yoke-Chen , Wang James D , Hahn Rita A , Gordon Marion K , Gerecke Donald R 
Published: June 2021

The gap‐junction proteins, the connexins, mediate intercellular communication and play a significant regulatory role in wound repairs. Recent studies showed that transient inhibition of connexin43 (Cx43) expression resulted in accelerated wound healing in both incisional and burn skin wounds. Skin wounds caused by vesicants generally result in an acute immune response and delayed wound repairs. We tested Cx43 antisense oligodeoxynucleotides (asODN) in a time course study of SKH‐1 mouse skin injury induced by the vesicant, nitrogen mustard, to observe the wound repair response. Animals treated with asODN had a significantly improved survival rate after 10 days when compared to all other control groups. Histological examination of tissue sections showed less acute inflammation for the asODN treated group when compared to the control groups. RT‐PCR and Western blot analysis showed a reduction of Cx43 for days 1 and 3. Cx 26 and 30 were also reduced at days 1 and 3 as shown by RT‐PCR. IL1B, the proinflammatory cytokine is five times less in the asODN samples compared to NM alone. IL10, the anti‐inflammatory cytokine is three fold higher compared to NM alone at three days post exposure. Macrophage elastase decreased at days 7 and 10 when compared to NM alone. This suggests Cx43 is essential to wound repair signaling. The potential use of Cx43 asODN helped reduction in secondary damage and a promotion of vesicant wound repair.Grant Funding Source: ES005022, ES004738, EY09056, and NIAMS U54AR055073
CKD712 promotes wound closure by producing VEGF through AMPK‐dependent HO‐1 induction
Authors: Jang Hwajin , Park Eunjung , Kim Hyejung , Chang Kichurl 
Published: June 2021

Given the importance of vascular endothelial growth factor (VEGF) and heme oxygenase (HO)‐1 in wound healing, the present study was tested our hypothesis that CKD712, a strong HO‐1 inducer, produces VEGF through HO‐1 induction for stimulating wound healing. Results indicated that CKD712 significantly increased VEGF production and accelerated migration of human dermal fibroblast (HDF) where addition of anti‐VEGF antibody antagonized the effect of promoting cell migration. Both AMPK inhibitor (compound C) and HO‐1 inhibitor (SnPPIX) but not inhibitors of MAPKs, PI3K, and PKC significantly reduced the production of VEGF by CKD712. Interestingly, SnPPIX inhibited HO‐1 expression but not p‐AMPK by CKD712. By contrast, compound C inhibited both p‐AMPK and HO‐1 induction by CKD712, indicating that AMPK controls HO‐1 induction. Moreover, CKD712 decreased HO‐1 induction without affecting the expression of p‐AMPK in siHO‐1 transfection but it failed to induce HO‐1 induction in siAMPKƒÑ1‐ transfected cells, supporting that AMPK is involved in HO‐1 induction by CKD712. In addition, administration of CKD712 significantly shortened the time of wound closure in SnPP IX‐sensitive manner in full‐thickness skin wound mice model. Taken together, we concluded that CKD712 accelerates cutaneous wound healing, at least, by production of VEGF through AMPK‐dependent HO‐1 induction.Grant Funding Source: NRF (03‐2010‐0298)