bluuebunny/crossref_metadata_2025_split huggingface dataset:
5‐HT2B Receptor Expression in the Phrenic Motor Nucleus of Rats with Intrapleural CTB‐Saporin‐Induced Phrenic Motor Neuron Death
Authors: Borkowski Lauren F. , Nichols Nicole L. 
Published: June 2021

Intrapleural injection of cholera toxin B fragment conjugated to saporin (CTB‐SAP) selectively kills respiratory motor neurons, and mimics aspects of neuromuscular disorders and neurodegenerative diseases including phrenic motor neuron death and decreased phrenic motor output. In this model, respiratory plasticity (acute intermittent hypoxia induced phrenic long‐term facilitation; pLTF) is enhanced following 7 days (d) of CTB‐SAP compared to control treatment. In contrast, pLTF in 28d CTB‐SAP treated rats is comparable to pLTF in controls, suggesting that the underlying mechanisms of plasticity at both time points may differ. pLTF in naïve rats predominately requires 5‐HT2 receptors; however, there is an alternative mechanism for pLTF that requires A2A receptors. Our preliminary data suggest that A2A receptors are increased in the phrenic motor nucleus of 7d, not 28d, CTB‐SAP treated rats (Craig et al., 2017), suggesting that 7d CTB‐SAP rats utilize the alternative mechanism for enhanced pLTF. However, it remains unknown how CTB‐SAP induced phrenic motor neuron death affects 5‐HT2A/B receptor expression, and if 5‐HT2A/B receptors are utilized for pLTF in CTB‐SAP treated rats. Here, we tested the hypothesis that 5‐HT2B receptor expression in the phrenic motor nucleus is decreased in 7d CTB‐SAP treated rats, but is not different in 28d CTB‐SAP treated rats when compared to controls using immunohistochemistry. 5‐HT2B receptor expression was studied in Sprague Dawley rats 7 and 28 days following bilateral intrapleural injections of: 1) CTB‐SAP (25μg), or 2) un‐conjugated CTB and SAP (control). Our preliminary data indicate that following CTB‐SAP injection, total 5‐HT2B receptor positive pixels and C4 fractional area are decreased in the phrenic motor nucleus and non‐phrenic ventral horn at 7d, not 28d, vs. controls (n=3 for each group; p<0.05). This suggests 5‐HT2B receptors recover to control levels and are potentially utilized for pLTF in 28d CTB‐SAP treated rats. Future studies will be focused on whether 5‐HT2A receptor expression is similarly affected, whether 5‐HT2A/B receptor expression is impacted on other cell types (i.e., glial cells), and which 5‐HT receptor isoform is required for pLTF following CTB‐SAP‐induced phrenic motor neuron death. This project increases our understanding of the expression of receptors associated with respiratory plasticity and its implications for breathing following motor neuron death.Support or Funding InformationSupported by NIH K99/R00 HL119606This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Extracellular Vesicles: An Additional Compartment for the Second Messenger, cyclic Adenosine Monophosphate (cAMP)
Authors: Sayner Sarah L. , Maulucci Marcy , Scruggs April , Zhou Chun , Bauer Natalie 
Published: June 2021

RationaleCyclic AMP is generated by adenylyl cyclase to initiate intracellular signaling leading to diverse cellular and tissue responses. Compartmentalization of the cAMP signal within the cellular environment is critical to maintain signaling specificity; however, cAMP is also released into the surrounding extracellular space. Many cell types release extracellular vesicles that encapsulate a variety of functional proteins and RNAs/miRNAs; however, little is known about whether they encapsulate signaling molecules, such as cyclic adenosine monophosphate (cAMP), which could contribute to the extracellular cAMP pool.MethodsTo test the hypothesis that endothelial cells release extracellular vesicles containing cAMP, media was collected from unstimulated aortic and pulmonary endothelial cells and subject to low‐speed and subsequently high‐speed centrifugation. To determine whether cAMP in these extracellular vesicles increased upon cell stimulation, pulmonary microvascular endothelial cells (PMVEC) were stimulated over time to increase cellular cAMP and extracellular vesicles analyzed for cAMP levels. Finally, isoproterenol and rolipram was perfused through the isolated rat lung to determine whether extracellular vesicles isolated from the intact organ also contain cAMP.ResultsWhile cAMP was detected in extracellular vesicles derived from various endothelial cell types, cAMP was higher in PMVECs compared to systemic endothelial cells (aorta) or endothelial cells from pulmonary conduit vessels, pulmonary artery endothelial cells. Further, stimulation of PMVECs with agents that increase near membrane cAMP (isoproterenol or forskolin) in the presence of the phosphodiesterase inhibitor, rolipram, led to an increase in cAMP in extracellular vesicles over time (upto 60 minutes). In PMVECs, cell lysates showed a maximum increase in cAMP 15 minutes after treatment, while the maximum increase in extracellular vesicle‐cAMP was not observed until 60 minutes after treatment. Extracellular vesicles from the perfusate of isoproterenol and rolipram stimulated isolated lungs contained elevated cAMP compared to unstimulated controls.ConclusionExtracellular vesicles released from both the systemic as well as pulmonary vasculature contain cAMP, and cAMP within extracellular vesicles can be increased upon cell stimulation. While cAMP is rapidly packaged into extracellular vesicles within 5‐minute of cell stimulation, the maximal increase is delayed. Isolated lungs can also be stimulated to release extracellular vesicles with increased cAMP. Thus, extracellular cAMP while already accepted to be free in the cytosol is also encapsulated within extracellular vesicles.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evidence for Altered Non‐Canonical Wnt Signaling and Increased Tight Junction Remodeling in Cftr Knockout (KO) Mouse Small Intestine
Authors: Clarke Lane L. , Woode Rowena A. , Liu Jinghua L. , Walker Nancy M. , Strubberg Ashlee M. 
Published: June 2021

Recent investigations have shown Cftr expression and its functional activity in the Lgr5+ intestinal stem cell (ISC) population of mice where it acts as a negative regulator of proliferation (Strubberg et al., CMGH 2017). In freshly‐isolated small intestinal crypts and crypts of cultured enteroids from Cftr KO mice, loss of Cftr activity results in increased Wnt/β‐catenin signaling and ISC hyperproliferation. A persistent alkaline intracellular pHi in Cftr KO ISC increases plasma membrane association of the Wnt transducer Disheveled (Dvl) thereby facilitating binding to the Wnt receptor Frizzled 7. Dvl also acts a transducer of non‐canonical Wnt signaling involved in directed cell migration and cell polarity, a necessary accompaniment to crypt proliferation. It is known that both cystic fibrosis (CF) patients and Cftr KO mice exhibit increased intestinal permeability. We hypothesized that increased pHi in progenitor cells of the crypt transit‐amplifying (TA) zone in Cftr KO mice increases plasma membrane Dvl association, potentially driving non‐canonical Wnt signaling for active tight junction (TJ) remodeling and increased permeability. Dvl2 association at the apical membrane was measured in TA cells of live WT and Cftr KO enteroids from mice expressing Dvl2‐EGFP (Dvl2 KO/Dvl2‐EGFP rescue) and membrane Tomato (RosamT/mG) using fluorescence confocal microscopy. Activity of the noncanonical Wnt receptor Frizzled 6 (Fz6) and the Rho GTPase Cdc42, an important regulator of intestinal polarity and migration, assessed TJ remodeling. Enteroid permeability was measured by FITC‐dextran (3 kD) diffusion into the lumen. Dvl2‐EGFP intensity at the apical membrane in TA zone crypt cells was increased ~2‐fold in Cftr KO crypts as compared to WT. Increased Dvl2‐EGFP apical membrane localization could be normalized in Cftr KO crypts by two days exposure to reduced medium pH (pH 6.6). Immunolocalization indicated the Fz6 was well expressed on the basolateral membranes near TJ of both WT and Cftr KO TA zone cells. Immunolocalization revealed increased Cdc42 activity at the TJ and immunoblots showed increased Cdc42 protein amounts in both fresh and enteroid crypts in Cftr KO intestine as compared to WT. Intestinal permeability was significantly increased in Cftr KO vs. WT enteroids. We conclude that cell alkalinity in the Cftr KO crypt epithelium affects Dvl transduction of both canonical and non‐canonical Wnt signaling thereby abnormally increasing proliferation and TJ remodeling. Increased TJ remodeling may contribute to increased intestinal permeability in CF patients.Support or Funding InformationSupported by NIDDK 5R01DK048816 and the Cystic Fibrosis Foundation (CLARKE11G0 and LIU13Q0).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Prorenin receptor regulates lipid metabolism via distinct actions on PPARγ and SREBP‐2 in liver of male mice
Authors: Gatineau Eva o. , Yiannikouris Frederique 
Published: June 2021

Introduction and objectivesThe epidemic of obesity is currently affecting more than 35% of the population in the United States. Non‐alcoholic fatty liver disease (NAFLD) is strongly associated with obesity and metabolic syndrome. Our laboratory previously demonstrated that adipose prorenin receptor (PRR) KO mice developed lipodystrophy and liver steatosis associated with elevated hepatic triglycerides (TG) levels. Interestingly, hepatic PRR and PPARγ expression were up‐regulated in HF‐fed adipose PRR KO mice. Since body of evidence indicated that PPARγ becomes elevated in fatty livers and since we showed that PRR regulated PPARγ in adipose, we sought to investigate whether PRR regulated TG contents in liver through a PPARγ‐dependent mechanism.Methods and resultsPRR floxed male mice fed a standard diet were injected with a single dose of an adeno‐associated virus thyroxine‐binding globulin Cre (Liver PRR KO, n=5) or saline (Veh, n=6). Body weights were measured weekly for 3 weeks. The deletion of PRR in liver did not change body weight curves. Hepatic levels of TG were significantly decreased in liver PRR KO mice compared with control mice injected with saline (Veh, 53.22 ± 15.4 mg/g liver; AAV, 10.79 ± 1.61 mg/g liver, P<0.05). The deletion of PRR induced a significant decrease of PPARγ gene expression whereas SREBP‐1c expression was unchanged suggesting that PRR regulated TG contents through a PPARγ‐dependent mechanism. Surprisingly, liver weights were significantly higher in liver PRR KO mice compared to control mice injected with saline (Veh, 1.82 ± 0.12 g; Liver PRR KO, 2.37 ± 0.08g, P<0.05). Plasma cholesterol levels (Veh, 143 ± 10 mg/dL; Liver PRR KO, 383 ± 14g mg/dL, P<0.05) and hepatic total cholesterol levels (Veh, 3.64 ± 0.09 mg/g liver; Liver PRR KO, 10.47 ± 0.55 mg/g liver) were elevated in liver PRR KO mice. Interestingly, the increase in total cholesterol content in liver was associated with an increase in SREBP‐2 and HMGCoA‐reductase gene expression suggesting that the deletion of PRR stimulated cholesterol synthesis. Additionally, ABCG5 gene expression was increased, reflecting an increased cholesterol efflux.ConclusionTaken together our results demonstrated for the first time that PRR is an important modulator of lipid metabolism in liver. In agreement with our hypothesis, hepatic PRR positively regulated TG contents through a PPARγ‐dependent mechanism. Interestingly, the deletion of hepatic PRR stimulates cholesterol synthesis via SREBP‐2 up‐regulation indicating that PRR acts at the crossroad between triglycerides and cholesterol metabolism.Support or Funding Information

COBRE ‐ P20 GM103527
American Heart Association ‐ 13SDG17230008
University of Kentucky, Center for Clinical and Translational Sciences, CCTS pilot grant, CCTS small grant, NIH T32HL091812
This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparative study of the intestinal microbiota in children from Mexican mothers with or without gestational diabetes
Authors: de Leon‐Solis Andrea Garcia , Torres‐Trueba Mariana , Palacios‐Gonzalez Berenice , Aguilar‐Reyes Jaqueline , de Leon‐Mendez Maria del Carmen Garcia , Lara‐Anaya Monica , Vadillo‐Ortega Felipe , Meraz‐Cruz Noemí 
Published: June 2021

Pregnancy includes the development of an individual, from fertilization to labor. This development is influenced by genetic and environmental modifications during the intrauterine period, which continues until the first 1000 days of life. Important lapse in health and disease programming. The presence of gestational diabetes (GDM) favors the development of metabolic diseases in adult life and has an 18% prevalence in Mexico. The diversity of the intestinal microbiota is affected by maternal diabetes status, and this could contribute to the abundance of different phylas that contribute to the generation of new associated diseases. The aim of this study was to compare the fecal microbiota of children between 3 months and 3 years of age born to mothers with and without GDM. Anthropometric measures and fecal sample were collected from 29 patients (without GDM=24; GDM=5). The V4 region of bacterial 16S rRNA gene was sequenced by Illumina MiSeq. There were no significant differences between the birth weight of the infant and the weeks of gestation. Our data showed differences in composition of the microbiota in the study groups. We observed a decrease in Firmicutes, in addition with an increase in Bacteroidetes and Proteobacterias in GDM group. Also, LefSe analysis indicated a significant increment in Desulfovibrionaceae in this group compared with the control group. These results suggest the presence of dysbiosis in child from gestational diabetic mothers.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Beta‐hydroxy‐beta‐methylbutyrate (HMB) augments muscle contraction‐induced protein synthesis via mTORC1 signaling in cultured L6 myotubes
Authors: Sato Satoko , Takamura Yusuke , Yamana Ikko , Nomura Mitsuru , Uchiyama Akira , Furuichi Yasuro , Manabe Yasuko , Fujii Nobuharu L. 
Published: June 2021

IntroductionBeta‐hydroxy‐beta‐methylbutyrate (HMB), a metabolite of L‐leucine, has been shown to enhance resistance exercise‐induced protein synthesis in human and animal studies. Since there is no suitable contraction model using skeletal muscle cell line for analyzing protein synthesis level, the combined effects of HMB and muscle contraction on the protein synthesis is still unknown.ObjectiveThe present study aimed to establish a new muscle contraction model in L6 myotubes for analyzing protein synthesis. Using this model, we investigated the effects of HMB on the contraction‐induced protein synthesis via the mammalian target of rapamycin complex 1 (mTORC1) pathway.MethodsL6 myotubes seeded into 4‐well rectangular plates were placed in contraction system equipped with carbon electrode at each end. The myotubes were stimulated with electric pulses of 50 V at 100 Hz for 10 min followed by the HMB treatment. Cells were collected at 3 hours poststimulation and evaluated mTORC1 signaling pathway activation by Western blot analysis.ResultsThe phosphorylation level of Akt which is upstream components of mTORC1 pathway and the phosphorylation of 70‐kDa ribosomal protein S6 kinase (p70S6K) which is downstream components of this pathway were significantly increased 3 hours after electric stimulation (p<0.05). Furthermore, the phosphorylation level of p70S6K in contraction cells combined with HMB was significantly increased compared with that in contraction‐only cells (p<0.05).ConclusionWe established a new muscle contraction model for analyzing protein synthesis in cultured L6 myotubes. Additionally, we found that HMB enhances muscle contraction‐activated mTORC1 signaling pathway using this model.Support or Funding InformationThis work was funded by Lion Corporation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cardiorespiratory Fitness Discriminates Sarcopenic (Abdominal)Obese from Sarcopenic Aging Subjects
Authors: Burini Roberto Carlos , Manda Rodrigo Minoru , Teixeira Okesley 
Published: June 2021

The present study aimed to characterize sarcopenic obesity (SpOb) and its related morbidities in free‐living adults clinically selected for a lifestyle modification program(LiSM).The study analyzed retrospectively baseline data from 523 individuals, of both genders that spontaneously joint the LiSM (Move for Health_Mexa‐se pro‐saude) program. The selected sample had data of medical anamneses, anthropometry, physical (and cardiorespiratory) fitness and blood biochemistry. Abdominal obese (Ob) definition, was defined by altered waist circumference (NCEP‐ATP III). Sarcopenia (Sp) was defined by the lower quartile of muscle‐mass index (kg muscle mass/m2 height=MMI). Statistical analyzes were performed by SAS software 9.2 (p <0.05).The sample was 54.74 ± 10.18 years old, 72.7% females, 28.5% sarcopenic, 69.6% (abdominal) obese and 20.6% SpOb. Ob subjects presented higher BMI, % BF and MMI, along with higher values of triglycerides, blood glucose, HOMA‐IR, CRP, uric acid and reduced values of plasma HDL‐c, trunkal flexibility and VO2max. Sp was associated with lower values of handgrip strength, HDL‐c and higher HOMA‐IR. SpOb Individuals were older and presented lower muscle strength and aerobic fitness as well lower plasma HDL‐c and higher HOMA‐IR. After adjustments cardiorespiratory fitness was the only age‐dependent discriminant of (abdominal) obese sarcopenia.Thus, The anthopometric diversion of SpOb keeps the pro‐inflammatory status (higher HOMA‐IR and lower HDL‐c) of its components (Ob and Sp) and has cardiorrespiratory fitness, as the sole aging‐dependent discriminant.Support or Funding InformationSupport CNPq and CAPESThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Shrinking the Mystery of Tsantsa Identification: Using Qualitative and Quantitative Measurements to Distinguish Natural and Imitation Shrunken Heads
Authors: Vinovskis Carissa , Oya Mayu , Royer Danielle 
Published: June 2021

INTRODUCTIONDuring the 17th through mid‐20th centuries, the Jivaro tribes of South America participated in ritual head shrinking as part of a religious ceremony following battle. In the late 19th and early 20th centuries, travelers to South America took interest in these shrunken heads, known as tsantsas, and brought them back home as curios, inspiring the production of fake shrunken heads for the tourist trade. Numerous tsantsas, both real and fake, have ended up in museums or private collections worldwide. As many museums have very specific rules about repatriation of human remains, understanding differences between authentic and imitation tsantsas is imperative.GOALSThe purpose of this study was to qualitatively and quantitatively describe the five tsantsa specimens held in the Anthropology Collection curated at the Denver Museum of Nature and Science (DMNS).METHODSFirst, each specimen was scored independently by two observers for 19 qualitative features typically associated with authentic tsantsas. Each observer also collected 29 standard facial measurements from four key locations (eye = 6, nose = 9, mouth = 4, ear = 10). Each facial measurement was size‐corrected using the geometric mean of all measurements for the specimen. The size‐corrected facial measurements were analyzed using principle component analysis (PCA).RESULTSInter‐observer agreement on scoring of the qualitative features was high, and analysis of the facial measurements revealed no significant differences between raters. The qualitative features suggested the presence of two distinct groups within the DMNS sample: A Probably‐Authentic Group (three specimens) exhibiting 15 or more out of the 19 authentic characteristics, and a Probably‐Imitation Group (two specimens) exhibiting 8 or fewer of the 19 characteristics. PCA of all size‐corrected variables identified two factors accounting for 78% of the variance in the sample (factor 1 = 47%; factor 2 = 31%). The PCA revealed that the five specimens clustered within two distinct groups, distinguished primarily along factor 1. The group composition determined by PCA was concurrent with the two groups indicated by the qualitative traits.CONCLUSIONSAgreement between analysis of qualitative tsantsa characteristics and facial morphometric measurements suggests the presence of three authentic and two imitation tsantsas in the DMNS collection. To affirm these findings, microscopic hair analysis is also performed to establish the presence of human hair. However, DNA analysis is needed to determine with confidence whether the DMNS tsantsas are human or fabricated.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Allometry of Felid Knee and Elbow Effective Mechanical Advantage
Authors: Harper Christine M. , Sylvester Adam D. 
Published: June 2021

Body size has a profound impact on animal locomotion because all terrestrial animals must support and move their mass against gravity. If large animals were isometrically scaled versions of smaller animals, then larger animals would have relatively weaker muscles and bones, as both are a function of area. The loads associated with locomotion however, are largely equivalent to body mass, a function of volume. To contend with this problem, larger animals have been observed to use more extended limb postures; increasing the effective mechanical advantage (EMA) of their muscles. Felids however, maintain the same limb posture across body sizes, suggesting they are not increasing EMA through limb extension. To investigate how felids maintain the same posture across body sizes we analyzed the relationship between the EMA of the triceps brachii and quadriceps and body mass in nine species of felids with kinematic data from the literature.Ulnae (n=37) and femora (n=30) of nine species of felids were surface scanned using a 3D NextEngine laser scanner. Linear measurements were taken on the tibia, patella, talus, calcaneus, navicular, third cuneiform and third metatarsal. Ulna and femur surface models were aligned in virtual space using anatomical and biomechanical axes. Triceps brachii EMA was calculated as the ratio of the muscle lever arm to the GRF lever arm, which were both collected on the ulna surface model. The quadriceps muscle and GRF lever arms were calculated using the femur surface models, linear measurements of limb and foot bones and kinematic data. Joint EMAs, muscle lever arms and GRF lever arms were regressed against body mass using PGLS regressions incorporating a phylogeny from 10kTrees. Pagel's λ was zero for all analyses, so data were reanalyzed using reduced major axis (RMA) regressions without phylogenetic information.The triceps brachii and quadriceps EMA scale to body mass with slopes of 0.15 and 0.13, respectively, indicating positive allometry, meaning that larger felids have a relatively greater EMA and suggesting that some aspect of felid joint morphology is altered. For both joints neither the muscle lever arms nor the GRF lever arms are statistically different from isometry, however the muscle lever arms generally trend toward positive allometry, while the GRF lever arms lean toward negative allometry. This indicates that either there are subtle changes to both lever arms or that different felid species accomplish an increased EMA in distinct ways.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.