bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serotonin Type 3 Receptor B Subunit (5‐HT3BR) and Corticotropin‐Releasing Hormone (CRH) Expression in the Serotonin Transporter Knockout (SERT KO) Rat Model of Visceral Hypersensitivity
Authors: Mecca Andrew , El‐Ayache Nadine , Galligan James 
Published: November 2023

BackgroundVisceral hypersensitivity is a chronic abdominal pain symptom present in functional gastrointestinal disorders such as irritable bowel syndrome (IBS). IBS is a global health issue, and is more prevalent in females than in males suggesting a potential role of female sex hormones in its pathogenesis. Some drug treatments for IBS target the ionotropic serotonin type 3 receptor (5‐HT3R), but these drugs have limited clinical benefit and can be associated with serious side effects. Therefore, a deeper understanding of the pathophysiology of visceral hypersensitivity is crucial for the development of more effective treatments. Environmental stress and altered serotonergic signaling may influence the severity of visceral hypersensitivity in IBS patients. Corticotropin‐releasing hormone (CRH) is an important mediator of the neuroendocrine stress response, but its role in IBS needs to be further studied.Aim/MethodsWe studied the expression of CRH and the B‐subunit of the 5‐HT3BR in the lumbar spinal cord, colon and lumbar dorsal root ganglia (DRG), in a serotonin‐transporter knockout (SERT KO) rat model of visceral hypersensitivity using immunohistochemistry and real‐time quantitative PCR.ResultsFemale SERT KO rats have higher proportion of CRH+ neurons in DRG than male SERT KO rats (P < 0.01), and CRH was most strongly expressed by small‐diameter nerve cell bodies that are likely to be involved in the pain pathway. There was no difference in the distribution of 5‐HT3BRs in the dorsal spinal cord or colon between SERT KO and WT animals. Similarly, there was no difference in CRH+ neurons in the spinal cord, and no difference in CRH expression in the colon between SERT KO and WT. However, we found a 10‐fold and 6‐fold increase in CRH mRNA expression in female and male SERT KO rats, respectively, compared with wildtype controls (P < 0.05). Lastly, preliminary data show that there is no difference in the mRNA transcript for the 5‐HT3BR in DRG between SERT KO and WT rats.ConclusionsWe found a novel sex‐specific increase in CRH expression in female SERT KO animals at the level of primary afferent neurons at the protein and mRNA levels. We found no difference in the distribution of the 5‐HT3BR between SERT KO and WT in the colon, DRG and spinal cord. Our results support our hypothesis that environmental stress mediated by CRH may influence visceral hypersensitivity in IBS.
Analysis of Inflammatory Modulation on Term and Preterm Neonatal Gastrointestinal Non‐Human Primate Explants
Authors: Alana Nicholas B , Acevedo Steven J , Carr Nick J , Mustafa Shamimunisa B , Blanco Cynthia L , King Jonathan M 
Published: November 2023

BACKGROUNDDysregulation of inflammatory processes are associated with intestinal injury, necrotizing enterocolitis, and risk of perforation in the newborn. A reduction of Epidermal Growth Factor (EGF) and impaired function of the Epidermal Growth Factor Receptor (EGFR), have been demonstrated in the preterm gastrointestinal system compared to term. Prostaglandin E2 (PGE2) as well as EGF have independently been shown to have a role in the barrier maintenance, recovery from injury, and regulation of perfusion.OBJECTIVEEvaluate the regulation of EGF/EGFR/PGE2 pathway in a non‐human primate gastrointestinal explant. Additionally evaluating the roles of inflammatory and anti‐inflammatory agents on this pathway.DESIGN/METHODSTissue explants were created from gastrointestinal regions (ileum through colon) upon necropsy of term and preterm (67% gestation) non‐human primates. Explants were evaluated in conditions of control, pro‐inflammatory (TNF 10 ng/ml), and anti‐inflammatory (indomethacin 50uM) variables. Absolute and relative prostaglandin secretion (PGE2); mRNA expression of Prostanoid (EP) receptor subtypes, EGFR, EPS8, AKT1, AKT2, and MAPK1 were used as markers of EGF/PGE2 pathway regulation at 24 hours post exposure.RESULTSPGE2 secretion significantly increases in term colon compared preterm colonic secretion. In addition, the term gastrointestinal explants are significantly more resistant to indomethacin inhibition of PGE2 secretion, noted across ileum, cecum, and colon. There were no significant differences noted in EP receptor subtype expression. Preterm expression of EGFR and its downstream effectors are elevated when compared to term ileum explants. However, there is markedly more variability in response noted to TNF and indomethacin in the term explants when compared to preterm ileum explants.CONCLUSIONSA positive gradient of PGE2 secretion in the distal gastrointestinal regions is observed in term explants likely correlated to the advanced differentiation status compared to the preterm. In addition, the up regulation of EGFR along with overall increased expression of downstream targets in the preterm explant is consistent with the expected mitogenic status. However, the decreased response to TNF and indomethacin, suggest a limited ability to modulate inflammation.
Tachykinin Activation on Enteric Glia: A Novel Mechanism of Enteric Nervous System Dysfunction in Irritable Bowel Syndrome
Authors: Delvalle‐Dorta Ninotchska Michelle , Rivera‐Lopez Gretchen , Gulbransen Brian 
Published: November 2023

The Enteric Nervous System (ENS) is the major neural regulator of the gastrointestinal (GI) tract. Enteric circuits involve precise interactions between neurons and glia and abnormal ENS activity is considered a key contributor to gut motility disorders like irritable bowel syndrome (IBS). However, the role of neuron‐glia interactions in functional GI motility disorders is not understood. Tachykinins are important excitatory neurotransmitters in the ENS that contribute to IBS pathophysiology. In the spinal cord, substance P (SP) and neurokinin A (NKA) release by hyperactive neurons drives reactive gliosis. We hypothesize that the activation of enteric glia by tachykinins plays an important role in neuroinflammation during IBS. We tested our hypothesis using immunohistochemistry (IHC) and calcium imaging recordings in tissue from 5–8 week old C57BL/6 male mice. Our results show that both NKA and SP are present in varicosities that surround enteric glia in the myenteric plexus. We found that this population of glial cells primarily express neurokinin‐2 receptors (NK2R) (n=3 animals) which preferentially bind NKA over SP. NK1R labeling in the myenteric plexus was primarily neuronal. However, we could not exclude the possibility of expression on glia. We did not find evidence of NK3R expression in the myenteric plexus. We tested the breadth of responsiveness of myenteric glia to tachykinins such as SP and NKA in whole‐mount preparations loaded with the calcium indicator dye Fluo‐4. Our results show that enteric glia produce robust calcium transients in response to stimulation with NKA but not to SP (n=62 glial cells from 3 animals). To determine if tachykinin receptor activation contributes to neurodegeneration during inflammation, we stimulated enteric glia with either SP or NKA and measured neuronal packing density within myenteric ganglia. Our results show that treatment with NKA significantly reduces neuronal survival (p=0.0416, n=4 animals) while treatment with SP had no effect (p=0.7171, n=3 animals). In conclusion, our results provide strong evidence that the activation of glial NK2Rs by NKA is a novel mechanism of neuroinflammation in the ENS.Support or Funding InformationT32NS44928‐11 and RO1DK103723
Arsenic (As) and Copper (Cu) Serve as Cofactors to Produce Hepatotoxicity With Mitochondrial Damage, Double Strand DNA Breaks and Disruption of ATM Signaling
Authors: Chinnasamy Thirunavukkarasu , Sharma Yogeshwar , Gupta Priya , Gupta Sanjeev 
Published: November 2023

Despite increasing exposure of populations to As the role of cofactors in organ toxicity, e.g., reactive Cu ions, is poorly defined. Since As and Cu ions may combine to increase oxphos events and processes directing mitochondrial and cellular DNA damage, we studied metal toxicity in cultured adult and fetal human hepatocytes with IC50 concentrations of As in MTT assays. In even nontoxic conditions by itself Cu dose‐dependently worsened As toxicity. This worsening included alterations related to oxphos activity with mitochondrial and nuclear perturbations (JC‐1 dye assays, acridine orange nuclear stain, 8‐OHdG stain), double strand DNA breaks (Comet assay, γH2AX stain), and replicative stress including loss of cycling cells and/or arrest in G2/M (Ki‐67staining, flow cytometry). In addition, verapamil‐sensitive side populations denoting stem cells were depleted by As toxicity. Use of cells stably transduced lentivirally to express hATM promoter construct with tdT reporter showed that Cu exacerbated cellular DNA damage by perturbing ATM signaling pathways. To further establish whether Cu accumulation may also worsen As toxicity in vivo, we studied Atp7b−/− mice that model Wilson's disease (WD) and accumulate Cu in liver due to absence of hepatobiliary Cu excretion, along with their healthy littermates. After As was given i.p. for 10d, liver necrosis increased significantly in Atp7b−/− mice (H&E tissue staining, serum hyaluronic acid levels). Moreover, we noted increased ATM pathway‐dependent DNA strand breaks (Comet assay, γH2AX stain). This As and Cu toxicity was ameliorated in human hepatocytes cultured with an anti‐oxidant, indole‐3 propionic acid (IPA), as shown by MTT, JC‐1 and hATM promoter assays. Similarly, IPA decreased As toxicity in Atp7b−/− mice.ConclusionsCu ions markedly exacerbated As toxicity with mitochondrial and ATM pathway‐regulated DNA damage. These findings will help understand how toxic metals may alter physiological processes and contribute additionally in human diseases, e.g., WD with Cu toxicosis.
Cardiorespiratory Fitness as a Predictor of Intestinal Microbial Diversity and Distinct Metagenomic Functions
Authors: Estaki Mehrbod , Pither Jason , Baumeister Peter , Little Jonathan P , Gill Sandeep , Ghosh Sanjoy , Ahmadi‐Vand Zahra , Marsden Katelyn R , Gibson Deanna L 
Published: November 2023

Background/ObjectiveAn imbalance between harmful and beneficial bacteria in the gut, termed dysbiosis, has been associated with a variety of illnesses. In specific, reduced community diversity has been implicated in various conditions such as inflammatory bowel disease, colorectal cancer, and irritable bowel syndrome amongst others. The role of physical fitness in the context of gut microbial community in humans is currently not known. A recent study of elite athletes suggested a positive association between fitness and microbial diversity in humans, however due to extreme dietary differences amongst the groups, disentangling the respective influences of fitness versus diet was not possible. Hence, we analyzed the gut microbiota of healthy subjects with varying cardiorespiratory fitness (CRF) levels with similar diets in order to isolate the relationship between fitness and microbial composition.Methods22 males and 19 healthy females of similar age, BMI, and diets participated in this study. The subjects’ aerobic capacities were determined using a standard peak oxygen uptake (VO2peak) test, considered the gold standard of CRF. High‐throughput sequencing of the 16S rRNA of fecal microbiota were carried using the Illumina platform. Fecal short‐chain fatty acids (SCFA) were analysed using gas chromatography.ResultsThe primary findings from this study showed that VO2peak was positively correlated with gut species richness (P=0.01, B‐coefficient=5.35) and can predict over 20% of the observed variability, while keeping all other factors constant, including diet. Further, protein intake but not VO2peak was significantly associated with changes in beta‐diversity, correlating strongly with Bacteroides genus. VO2peak was a significant factor in explaining variability of the microbiomes predicted metagenomic functions, aligning positively with genes related to bacterial chemotaxis and motility, and negatively with lipopolysaccharide biosynthesis. Of all SCFA analysed, VO2peak was positively correlated with only butyrate, known for its role in promoting gut health.ConclusionsResults from this study show that CRF is correlated with increased microbial diversity in healthy humans and that the associated changes are anchored around a set of functional cores rather than specific taxa. The microbial profiles of those with high CRF levels favor the production of the beneficial SCFA, butyrate. As increased microbiota diversity and butyrate production is associated with overall host health, our findings warrant further research in the use of exercise prescription as an adjuvant therapy in combating dysbiosis‐associated diseases.Support or Funding InformationThis project was funded by the Natural Sciences & Engineering Research Council (NSERC) of Canada
Vitamin D Supplementation and DNA Methylation Patterns during Pregnancy and Lactation in Mothers and Infants
Authors: Anderson Cindy M. , Thiele Doria K. , Ralph Jody L. , Perley Danielle , Ohm Joyce E. 
Published: November 2023

The purpose of this study was to identify the temporal influence of maternal vitamin D supplementation during pregnancy and lactation on DNA methylation in mothers and infants. In this double blind randomized control trial, pregnant women planning to exclusively breastfeed were randomized to control (400 IU vitamin D3 daily, n=8) or experimental (3800 IU vitamin D3 daily, n=8) groups from 28 weeks gestation through 4 weeks postpartum. Genome‐wide DNA methylation (Illumina Infinium 450K) was analyzed in white blood cells from mothers at 28 weeks gestation (baseline), 2 days postpartum and 4 weeks postpartum and in their fully breastfed infants at birth and 4 weeks of age. Significant DNA methylation was determined by at least 20% difference in individual CpG methylation between control and experimental groups. In the experimental group, differential methylation in >400 individual maternal CpGs in 280 known genes across the genome was identified with methylation loss present in 2/3 of individual CpGs. Among the infants, differential methylation in 143 CpGs (91 genes) was identified with common methylation loss patterns as in the mothers. These findings provide evidence for the direct influence of vitamin D supplementation on DNA methylation in pregnant and lactating women, in utero on the fetus and via breastmilk in newborns.Support or Funding Information1P20GM104360‐01 National Institutes of Health, National Center for Research Resources (NCRR) Centers of Biomedical Research Excellence (COBRE). Center of Excellence in Epigenetics/Epigenomics of Development and Disease. COBRE Epigenomics Jumpstart Pilot Grant, Epigenomic gestational responsiveness to vitamin D Association of Women's Health, Obstetric and Neonatal Nursing (AWHONN). Maternal vitamin D supplementation to correct deficiency in mothers and breastfed infants.
Inner Ear Volume in Craniosynostotic Mice
Authors: Heitman Erin Ashley , Holmes Megan A , McNulty Margaret A , DeLeon Valerie Burke , Mussell Jason C 
Published: November 2023

Craniosynostosis is a congenital defect causing premature fusion of cranial sutures. These skull abnormalities can cause developmental delays, cranial venous hypertension, raised intracranial pressure, hearing impairments, and facial deformities. The temporal bone, specifically, plays a crucial role in the development in many of the features affected by craniosynostosis. The jugular foramen is an interosseous foramen formed by the temporal and occipital bones. Stenosis of the jugular foramen has been reported in individuals suffering from both syndromic and non‐syndromic complex (multi‐suture) craniosynostosis. Children with complex craniosynostosis also exhibit anomalous basicranial venous drainage in the form of persistent condylar and mastoid emissary veins uniting the sigmoid dural venous sinus with the suboccipital venous plexus and posterior auricular vein, respectively. The otic capsule is crucial for proper establishment of the sigmoid dural venous sinus during development. The petrous portion of the temporal bone is a significant branch point for dural venous sinuses and is altered in children with craniosynostosis syndromes. This study aimed to compare the volume of the otic capsule in syndromic craniosynostotic mice and their wild‐type littermates for possible effects on basicranial venous drainage. Using the otic capsule as a proxy for temporal bone volume and symmetry, we hypothesize that craniosynostosis causes a change in otic capsule volume throughout ontogeny that may also alter dural venous drainage pathways.The Fgfr2+/C342Y mouse model of Crouzon's syndrome along with wild‐type littermates were analyzed using serial micro‐computed tomography sections of whole heads at embryonic day 17.5 (E17.5), postnatal day 14 (P14), and postnatal day 42 (P42). Further, a subset of mouse crania were immersed in potassium iodine solution (Lugol's solution) to stain soft tissue structures. Immersion was performed to enable measurement of the unossified otic capsule of the earliest stages. Amira 3D analysis software was used to reconstruct three dimensional isosurface structures and compute volumes from complete two‐dimensional CT image stacks.While we did not find evidence of otic capsule volumetric differences in the earliest stages examined, significant differences between Fgfr2+/C342Y and WT littermates did emerge in the sub adult population (P42; p = 0.0455). Further investigations will examine the surface area of the jugular foramen, as well as, emissary foramina.
Deletion of Notch signaling in the developing mouse kidney results in a disruption of planar cell polarity in tubule epithelial cells
Authors: Kearns Donovan , Livingston Safia , Baybutt Richard , Heuvel Gregory Vanden 
Published: November 2023

Cux1 is the murine homologue of the Drosophila gene cut. In Drosophila, multiple genetic interactions between Cut and the Notch and Wingless signaling pathways occur during development. Cux1 is upregulated by Notch1 activation in rat kidney epithelial cells, and interacts with the groucho homologue Grg4 to regulate p27 expression in the developing kidney. To further evaluate the regulation of Cux1 by Notch signaling, we conditionally deleted RBPJ/k, a transcription factor required for canonical Notch signaling, in the developing ureteric bud and collecting duct using Cre‐recombinase (Cre) under the direction of the Hoxb7 promoter. We confirmed the Cre activity by crossing the RBPJ/kCD mice to the membrane‐targeted Tomato/Green Fluorescent Protein (mT/mG) double fluorescent reporter mouse strain (Gt(ROSA)26Sortm4(ACTB‐tdTomato,‐EGFP)Luo/J), which carries a loxP‐flanked Tomato cassette. Newborn RBPJ/kCD mice exhibited medullary and cortical cystic like dilatations. Postnatal day 14 (P14), RBPJ/kCD mice exhibited hydronephrosis and cortical dilatations. Analysis of planar cell polarity using anti‐phospho‐Histone H3 antibodies showed a disruption of spindle orientation in the collecting ducts of RBPJ/kCD mice. Taken together, these results suggest that Notch signaling is required for the normal development of the collecting duct system in the kidney, and that reduced Notch signaling may contribute to the development of renal cysts.Support or Funding InformationNational Institutes of Health
Uterine Endometrial Reprogramming during Early Pregnancy is Accompanied by Distinct Spatiotemporal Expression Patterns of Basement Membrane Components
Authors: Jones‐Paris Celestial R , Paria Sayan , Berg Taloa , Saus Juan , Bhave Gautam , Paria BC , Hudson Billy 
Published: November 2023

Uterine endometrium undergoes decidualization, a tissue reprogramming process, in response to blastocyst implantation in mice. During this process, stromal cells acquire epithelial‐like behaviors including expressing basement membrane (BM), a specialized form of extracellular matrix that influences epithelial cell identity and behaviors. The assembly and function of BM during decidualization is unknown. In this study, we investigated uterine expression and compartmentalized localization of BM components: classical components collagen IV and laminin, recently identified component peroxidasin, and a putative component Good pasture antigen‐binding protein (GPBP), in the mouse uterus during early pregnancy. Electron microscopic investigation revealed: 1) a smooth continuous layer of BM in between the epithelial and stromal layers of the preimplantation endometrium; and 2) interrupted, uneven, and thickened BM within the pericellular space of the post implantation decidua. Quantification of mRNA levels by qPCR showed changed in expression levels that were complemented by immunofluorescence localization of collagen IV, laminin, peroxidasin, and GPBP. Novel observations of BM‐associated and distinct expression patterns of the four components suggest collective pericellular functions as well as individual functions of the components in the peri implantation uterus and decidua.Support or Funding InformationThis work was supported by National Institutes of Health Grants F31 CA165787‐03 (to C.R.J.), R25 DK096999 (to B.G.H. for S.P. and T.B.), R01 HD044741 (to B.C.P.), and R01 DK018381 (to B.G.H). This study was also supported by RT2014‐2415‐1 / Prometeo II ‐2014‐048 (to J.S).
IMPACT OF BMP‐7 ON CELL MIGRATION IN THE MENISCUS
Authors: Taylor Victor , Hutchinson Ian , Danelson Kerry , Ferguson Cristin 
Published: November 2023

IntroductionThe meniscus is a C‐shaped fibrocartilaginous tissue that is essential for the normal biomechanical function of the knee. Damage or loss of the meniscus is associated with osteoarthritis. Inflammatory cytokines like Interleukin‐1 (IL‐1) inhibit repair. However, Bone Morphogenetic Protein‐7 (BMP‐7) showed great regenerative potential in cartilage and may help aid repair in meniscus. The effect of BMP‐7 on cell migration in the meniscus was examined to evaluate the potential of this agent to augment meniscus repair and tissue engineering applications.MethodsThe effect of BMP‐7 on fibrochondrocyte migration was examined using a model that interfaces a collagen type I sponge treated with BMP‐7, IL‐1, and BMP‐7+IL‐1 between porcine meniscus explants in a closed system held via a cloning tube. In this model, the growth factor treated sponge serves as the constant stimulus (or bait) for cell migration away from the media. Explants were incubated for 28 days. Afterward, tissues were embedded in paraffin and stained with 4′,6‐diamidino‐2‐phenylindole (DAPI) stain. Initial data represents n=7 for Control, n=8 for BMP7, n=4 for IL‐1, and n=4 for BMP‐7+IL‐1 groups.ResultsExplant meniscus tissue histology sections were divided into scaffold, middle, and peripheral zones relative to position next to the growth factor treated sponge, and cell count was performed in each zone. There were significantly more cells in the middle zone of the BMP‐7 group compare to the other groups (p < .05). In addition, there were significantly more cells in the peripheral zone of the BMP‐7+IL‐1 group compared to the peripheral zone of the BMP‐7 group (p < .005).ConclusionPreliminary results suggest that BMP‐7 may induce cell migration in meniscus cells. More studies will be done to better determine the effects of BMP‐7 on the meniscus.Support or Funding InformationInstitutional Research and Academic Career Development Awards (IRACDA) (NIH/NIGMS K12 PAR‐13‐290)