bluuebunny/crossref_metadata_2025_split huggingface dataset:
Systemic Treatment with Cannabinoid Receptor 2 Agonist to Treat Osteoporosis in a Rodent Model of SCI
Authors: Grayson Bernadette E. , Pride Yilianys , Sereduck Susan , Chade Alejandro , Grill Raymond , Tucci Michelle 
Published: April 2020

Osteoporosis is a skeletal disease resulting in a reduction of bone mineral density and bone mass with disorganization of trabecular bone architecture, leading to an increased incidence of bone fracture. Nearly all spinal cord injury (SCI) persons will develop osteoporosis following injury, and further up to 50% of all persons with SCI will sustain a fracture during their lives. The mechanism driving osteoporosis following SCI is elusive. The cannabinoid system modulation of bone metabolism through CB1/2 has been increasingly of interest for the preservation of bone mass and density in models of osteoporosis. Here we used early and sustained treatment with selective cannabinoid 2 receptor (CB2) agonist, HU‐308, in a mouse model of SCI to prevent and reverse SCI‐induced osteoporosis. First, we determined the efficacy of acute treatment with CB2 agonist, HU‐308, to stave off osteoporosis with treatment for 40 days post‐injury. In thoracic‐complete transected mice, the bone density was significantly reduced in comparison to injury‐naïve mice. In the distal tibia, HU‐308 was effective to preserve trabecular bone density as measured by Micro‐CT in comparison to SCI mice treated with vehicle (VEH) (P < 0.05) but less so in the proximal tibia. In the femur, 40 days post‐injury, there was no efficacy of HU‐308 to preserve bone density in SCI‐HU‐308 treated mice in comparison to VEH‐treated SCI. Histologic inspection supports substantial loss of trabecular bone. In order to determine if HU‐308 could reverse bone loss, we treated SCI mice with HU‐308, CB2‐agonist significantly after the onset of SCI‐induced osteoporosis (i.e., 3 months post‐injury) for 40 days; this resulted in profoundly improved bone mineral density in SCI mice treated with HU‐308 in the tibia and trends in the femur. Additionally, increased periosteal bone formation was observed in the femurs of SCI‐HU‐308 treated animals following the delayed treatment (P < 0.05) and absent in the SCI‐VEH treated animals. We also sectioned undecalcified bone and stained with Alizarin to demonstrate preservation and rebuilding of bone with HU‐308 in the acute and delayed treatment paradigms. Acutely, there appears to be preservation of the cortical bone and not the trabecular bone; the VEH‐treated mice showed loss of mineralization in both trabecular and cortical bone. Delayed treatment with HU‐308 shows bone formation with a higher degree of trabecular and cortical mineralization in comparison to VEH‐treated SCI mice. Taken together these data suggest that the cannabinoid system contributes to the pathophysiology of SCI‐induced osteoporosis and that CB2 agonism may be a viable therapeutic strategy to prevent or slow osteoporosis in SCI patients.Support or Funding InformationThis work is supported by Department of Defense Grant Award Number: W81XWH‐16‐1‐0349 to BEG.
Adaptation of Puromycin‐Associated Nascent Chain Proteomics to Arabidopsis thaliana and Pisum sativum
Authors: Noras Aleksandra Izabela , von Arnim Albrecht 
Published: April 2020

Translation is a biological function present in all forms of life that allows for the production of proteins from strands of messenger RNA (mRNA). In plants, such as Arabidopsis thaliana or Pisum sativum (garden pea), translation is tightly regulated because of the plants’ sessile nature and need to adapt to a variety of stresses. PuNCH‐P is a proteomic technique that offers a snapshot of the translational status of cells in both a noninvasive and time sensitive manner. It was previously developed for mammalian tissues but had not been adapted to plants. PuNCH‐P delivers an estimate of the rate of protein production using a technique complementary to ribosome footprinting or polysome loading. This method circumvents the need for in‐vivo labeling such as in SILAC or BONCAT methods, and also avoids high sequencing costs associated with ribo‐seq methods. Nascent peptides attached to ribosomes are isolated from a frozen tissue sample by centrifugation through a sucrose cushion. These peptides are then C‐terminally labelled with biotin‐puromycin. The incorporation of biotin allows for downstream peptide visualization on an immunoblot (Western Blot) using streptavidin‐HRP as a detection antibody. Peptides may also be captured with streptavidin coated beads. Here, we validate the PuNCH‐P method for Arabidopsis and peas. It is well known that heat shock causes an overall decrease in the translational activity of plants due to mechanisms such as the sequestering of cytoplasmic mRNAs into stress granules and the shift to translating heat shock proteins (HSPs). Here we present methods to efficiently label, capture and detect biotin‐labeled nascent peptides from plants. With heat treatment as a proof of concept, we show that puromycylation recapitulates the shift in the patterns of translation expected from polysome profiling. These data set the stage for the subsequent proteomic analysis of nascent peptides in plants.Support or Funding InformationNSF MCB 1546402
The curious case of ligamentum arteriosum: It is more than a ligament
Authors: Quaye Benedicta , Kummer Wolfgang 
Published: April 2020

IntroductionThe ligamentum arteriosum is generally considered to be a mere a remnant of the embryonic bypass (ductus arteriosus) from the pulmonary circulation to the aortic arch, obliterating soon after childbirth. This study set out to elucidate the morphology, innervation, and neurochemistry of this structure.MethodThe mediastinum of wildtype mice (C57BL/6J, n=5), reporter mouse strains indicating the expression of the nicotinic receptor subunit α3 (chrna3‐eGFP, n=4) and of choline acetyltransferase (chat‐eGFP, n=4), human (n=10) and pig (n=9) ligaments obtained from body donors (4 male, 6 female, aged 69–97 years) and commercial abattoir, respectively, were subjected to single‐ and double‐labeling immunofluorescence using antibodies against structural markers such as α‐smooth muscle actin (αSMA), protein gene product 9.5 (PGP 9.5), neurofilament 68 (NF68), against neuropeptides such as neuropeptide Y (NPY), vasoactive intestinal peptide (VIP), substance P (SP), and calcitonin gene‐related peptide (CGRP), and against transmitter synthesizing enzymes such as tyrosine hydroxylase (TH; nordrenergic), choline acetyltransferase (ChAT; cholinergic), and nitric oxide synthase (NOS). Paraffin embedded human samples were stained with hematoxylin and eosin, Masson‐Goldner trichrom and elastica‐van Gieson dyes.ResultsIn mice and human ligaments, remnants of a lumen and of an internal elastic membrane were observed. The lumen of the “ligament” was not completely obliterated, even though it was not patent from one side to the other. Contrary to a canonical ligament, the ligamentum arteriosum is mainly made up of collagen and abundant αSMA‐positive cells in all three species. Some human samples contained a fibrous core, sometimes calcified. Blood vessels and nerve bundles were abundant in the adventitia. In mice, the point of attachment of the ligament to both the aortic arch and pulmonary trunk received a noticeable amount of chrna3‐eGFP+ but no cholinergic innervation. PGP 9.5‐, TH‐, NPY‐, CGRP‐, and SP‐immunoreactive nerve fibers were observed in this rank order of frequency. In pig, VIP+ fibers were additionally observed. In human ligaments, PGP 9.5+, Nf68+, TH+, NPY+, VIP+, CGRP+, and SP+ fibers were observed.ConclusionThe so‐called ligamentum arteriosum retains a continuous smooth muscular wall until senescence with noticeable, presumably postganglionic sympathetic (TH+, noradrenergic) and to a lesser extent sensory C‐fiber innervation (CGRP+, SP +) long after childbirth. It is clearly not a passive structure and the dense innervation could influence impedance/compliance (reported to be the best prognostic parameter for patient survival in pulmonary hypertension (Hunter et al., Am Heart J. 155:166–74, 2008) of the vascular segments to which it is attached.
Modulation of the tongue muscle activity in response to bronchopulmonary C‐fiber activation following mid‐cervical contused in the rat
Authors: CHANG HSIAO-SEN , LEE KUN-ZE 
Published: April 2020

The present study was designed to investigate the tongue muscle activity in response to bronchopulmonary C‐fiber activation following mid‐cervical spinal contusion in the rat. The esophageal pressure and electromyogram of extrinsic tongue muscles (genioglossus and hyoglossus) were monitored before and after inhalation of capsaicin (25 and 100 μg/ml) at the acute, subchronic and chronic injured stages after mid‐cervical contusion. The results demonstrated that pre‐inspiratory and inspiratory burst amplitude of tongue muscle were generally enhanced at the acute and subchronic injured stages in mid‐cervical contused animals vs. uninjured animals. Inhalation of capsaicin usually caused a rapid shallow breathing pattern associated with a reduction in pre‐inspiratory and inspiratory activity of the tongue muscle in uninjured animals at the acute injured stage; however, this altered breathing pattern and decreased tongue muscle activity were blunted in contused animals. During the chronic injured stage, the expiratory duration was prolonged to a similar degree in both uninjured and contused animals after inhalation of capsaicin. Notably, inhibition of genioglossus muscle activity during capsaicin inhalation is more severed in chronically contused animals. These results demonstrated that cervical spinal cord injury is associated with alteration of supraspinal respiratory activity, which may modulate upper airway motor outputs and bronchopulmonary C‐fiber mediated respiratory reflexes. The compensatory increase in the tongue muscle activity at the acute injured stage may be benefit to the maintenance of the upper airway patency; however, the susceptible tongue muscle activity during bronchopulmonary C‐fiber activation may increase the risk of airway obstruction during the chronic injured stage.Support or Funding InformationMOST (105‐2628‐B‐110‐002‐MY3 & 108‐2636‐B‐110‐001), Higher Education Sprout Project (07C030111), NSYSU‐KMU Joint Research Project (107‐I001)
The Effects of C‐Phycocyanin and Lycium barbarum Polysaccharides on Aspirin‐Induced Apoptosis in Rat Gastric Mucosal Cells
Authors: Chao Jane C-J , Liu Yu-Chen , Hsu Chien-Yeh , Chen Yu-Shan 
Published: April 2020

Non‐steroidal anti‐inflammatory drugs (NSAIDs) cause gastrointestinal injury through both topical and systemic effects, which may generate reactive oxygen species and lead to apoptosis. Aspirin, a NSAID, is widely used for cardio‐protection and causes gastrotoxicity in long‐term administration. C‐phycocyanin (CPC) is a biliprotein pigment of the blue‐green alga Spirulina platensis, and possesses antioxidant, anti‐inflammatory, and wound‐healing properties. Lycium barbarum polysaccharides (LBP) extracted from wolfberry act as an antioxidant and immunomodulator. This study investigated the effects of CPC and LBP on aspirin‐induced apoptosis in rat gastric mucosal RGM‐1 cells. Rat RGM‐1 cells were pretreated with different concentrations of CPC and/or LBP (100, 250, and 500 μg/mL) for 24 hours, and gastric damage was induced by 21 mM aspirin for 3 hours. Apoptotic protein expression was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting. The results showed that aspirin significantly increased c‐Jun N‐terminal kinase (JNK), p38 phosphorylation, and Bax expression, while LBP decreased JNK activation and Bax expression compared with aspirin treatment alone. However, CPC had no effects on these apoptotic markers. In conclusion, LBP may decrease aspirin‐induced apoptosis in rat gastric mucosal RGM‐1 cells by suppressing the activation of JNK signaling pathway.Support or Funding InformationThe study was supported by the Ministry of Science and Technology, Taiwan (grant no. MOST105‐2320‐B‐038‐036‐MY3).
Cardiovagal Baroreflex Sensitivity is Reduced in Middle‐Age and Older Men with Low Testosterone
Authors: Babcock Matthew C. , Cornwell William K. , Witten Teresa L. , Brubaker Ashley , Hildreth Kerry L. , Moreau Kerrie L. 
Published: April 2020

BACKGROUNDMale aging is associated with a general decline in serum testosterone (T) concentrations. Endogenous low T is associated with increased cardiovascular disease risk. Following orchiectomy or androgen suppression, rats have reduced cardiovagal baroreflex sensitivity (BRS), a key antecedent to the development of hypertension and other cardiovascular diseases. BRS is determined by mechanical (i.e. arterial stiffness) and neural factors, and heart rate variability (HRV) may provide insight into the contribution of these mechanical and neural factors. However, the effects of low T on BRS in middle‐age and older men is unknown.PURPOSEDetermine whether BRS and HRV are reduced in middle‐age and older men with low T.METHODSEight healthy older men between the ages of 50–75 years (BMI: 28.7±6.3 kg/m2, systolic blood pressure: 130±9 mmHg, diastolic blood pressure: 78±7 mmHg; mean±SD) with either low (<300 ng/dL; n=4) or normal (≥400 ng/dL; n=4) T underwent simultaneous recordings of blood pressure (finger photoplethysmography) and R‐R interval (electrocardiogram) during supine rest. Spontaneous cardiovagal BRS was assessed in sequences of ≥4 cardiac cycles in which R‐R interval and systolic blood pressure changed in the same direction. Time (standard deviation of R‐R intervals of normal sinus beats; SDNN, and root‐mean‐square of successive differences in R‐R interval differences; RMSSD) and
frequency (low frequency and high frequency power) domain indices of HRV were determined during six minutes of paced breathing at 0.2 Hz. Serum T was measured in the morning under fasting conditions.RESULTST concentrations were significantly lower in the low T group compared to the normal T (269.3±40.5 vs. 561.0±109.8 ng/dL; p=0.002). Age (56±3 vs. 57±5 years; p=0.836) and mean arterial pressure (96±9 vs. 98±8 mmHg; p=0.794) were similar between groups. Cardiovagal BRS was significantly lower in the low T compared to normal T group (
8.2±3.1 vs. 16.2±4.2 ms/mmHg; p=0.021). Cardiovagal BRS was strongly correlated with T concentrations when considered as a continuous variable (R=0.71, p=0.049), but cardiovagal BRS was not correlated to age (R=0.06, p=0.891). No indices of HRV were significantly different between groups (p≥0.20 for all).CONCLUSIONSThese preliminary data indicate that spontaneous cardiovagal BRS is reduced in men with low T. The absence of differences in HRV suggest that differences in BRS may be influenced by
mechanical (i.e. arterial stiffness), rather than neural mechanisms.Support or Funding InformationSupported by NIH Grants R01AG049762 and T32AG000279‐16A1
Kangxian‐Yixin Granule alleviates myocardial fibrosis in rats with dilated cardiomyopathy through RhoA/ROCK1 pathway
Authors: Wang Zhentao , Wang Hailiang , WU HONG , Han Lihua 
Published: April 2020

ObjectiveKangxian‐Yixin granule (KXYX) is an effective traditional Chinese medicine prescription for the clinical treatment of dilated cardiomyopathy (DCM). The purpose of this study was to study the role of RhoA/ROCK1 signaling pathway in DCM myocardial fibrosis and the intervention mechanism of KXYX.MethodsDCM rats were randomly divided into model group (n = 8), KXYX group (n = 8), captopril group (n = 8) and control group (n = 8). After 4 weeks of drug intervention, the cardiac function of rats was detected by echocardiography, and the degree of myocardial fibrosis was observed by Masson staining. The expressions of RhoA, ROCK1, p‐MLC and fibrosis related factors CTGF and Col‐1 in RhoA/ROCK1 signaling pathway were detected by PCR and Western blot. At the same time, the myocardial fibroblast model was induced by angiotensin II, and treated with KXYX and inhibitor fasudil. RhoA/ROCK1 signaling pathway and the expression of fibrosis factors CTGF and Col‐1 were detected by Western blot and PCR.ResultsCompared with the control group, the expression of RhoA/ROCK1 signaling pathway in the model group was up‐regulated, the expression of p‐MLC was overactivated, and the expression of fibrosis‐related factors CTGF and Col‐1 was increased in the model group (P < 0. 05). KXYX could inhibit myocardial fibrosis, inhibit the transformation of fibroblasts into myofibroblasts, and down‐regulate the expression of RhoA/ROCK1 signaling pathway and fibrosis factors CTGF and Col‐1 in DCM rats.ConclusionRhoA/ROCK1 signaling pathway plays an important role in DCM myocardial fibrosis, and KXYX can inhibit the up‐regulation of RhoA/ROCK1 signaling pathway, thus inhibiting the development of myocardial fibrosis in DCM rats. The correspondence to Lihua Han;
hlhaa@sina.com.Support or Funding InformationThis work was financially supported by grants from the National Natural Science Foundation of China (Grant No. 81573920)
G Protein Coupled Receptor 75 and Its Role in Pulmonary Hypertension
Authors: D’Addario Catherine , Kitagawa Atsushi , Zhang Frank , Zeldin Darryl , Gruzdev Artiom , Garcia Victor , Schwartzman Michal , Gupte Sachin 
Published: April 2020

Pulmonary Hypertension (PH) is a multifactorial disease with poor prognosis and inadequate treatment. Recent studies have shown that the orphan G Protein Coupled Receptor 75 (GPR75) is upregulated in the pulmonary vessels of PH patients, especially in females. Therefore, we hypothesized that GPR75 knock out (GPR75KO) mice will be protected from developing PH. We placed 3‐month‐old female GPR75KO (n=6) and control C57BL/C mice (n=10) in a hypoxic chamber with 10% Oxygen or at atmospheric oxygen for 5 weeks. At the end of 5 weeks, we performed echocardiograms and right heart catheterizations. GPR75KO mice were protected from developing PH (Right Ventricle Systolic Pressure (RVSP) of 26mmHg and Right Ventricle Diastolic Pressure (RVDP) of 2 mmHg), while C57BL/C mice developed PH under hypoxia (RSVP and RDVP of 56mmHg and 12mmHg, respectively). Wire myography was performed on isolated intra‐lobar pulmonary arteries from GPR75KO mice (n=3) and WT mice (n=31). Hypoxic pulmonary vasoconstriction (HPV) was abolished in GPR75KO as compared to WT. HPV response in WT was blocked (P<0.05) by antibody against RANTES (CCL5), a putative GPR75 ligand. These results suggest GPR75 plays a major role in the development of HPV and PH.Support or Funding InformationThis study was supported by NIH grant 1RO1HL139793 (MLS).
Effects of Arginine Vasopressin and Oxytocin on Neurite Outgrowth and Cellular Signaling in SH‐SY5Y Human Neuroblastoma Cell Line
Authors: Pierce Marsha 
Published: April 2020

IntroductionIn the central nervous system, arginine vasopressin (AVP) and oxytocin (OT) stimulate neural networks that regulate social behaviors, including social attachment, aggression, and complex social cognition. AVP and OT are neuropeptides that are expressed in the social behavioral network and bind to G‐protein coupled receptors, stimulating cellular signaling patterns that ultimately affect social behaviors at the organismal level. Consensus mammalian AVP and OT sequences are highly conserved. In addition to sequence homology between AVP and OT, there is ~85% structural homology between the OT receptor (OTR) and vasopressin 1a receptor (V1aR) resulting in significant cross‐reactivity between the ligands and receptors. Perturbations in OT and/or OT receptor expression results in social behavioral deficits, and is associated with a number of psychopathologies including autism spectrum disorder, schizophrenia, anxiety, and depression.MethodsThe human neuroblastoma derived SH‐SY5Y cell line demonstrates morphological characteristics of neurons including neurites and formation of functional synapses, endogenously expresses OTR, and is often used as an in vitro model for human neurons. Functional assays were performed using Fluo8‐AM to measure ligand‐induced Ca2+ mobilization and FLIPR Membrane Potential (FMP) assays were performed to assess ligand‐induced hyperpolarization. To assess effects on neurite outgrowth, cells were treated with logarithmic doses of AVP and OT for 24 hours, fixed and stained with phalloidin and DAPI.ResultsIn SH‐SY5Y cells, OT was more potent than AVP at both ligand‐induced Ca2+ mobilization and membrane hyperpolarization, which is consistent with OT being more potent and efficacious at the human OTR. Specificity of the signaling at the OTR were confirmed using the vasopressin 1a receptor antagonist SR49059 and the oxytocin receptor antagonist L‐371,257. These data are consistent with previous reports that vasopressin 1a receptors are absent in SH‐SY5Y cells. Additionally, OT demonstrated a dose‐dependent increase in neurite outgrowth, which is inhibited by L‐371,257.ConclusionsTogether, these data demonstrate that OTR mediated signaling pathways lead to neurite outgrowth in SH‐SY5Y cells. Additional studies are needed to assess the specific molecular mechanisms that contribute to neurite outgrowth, and whether functional synapses are formed. Integrative studies of behavior, genetics and ligand‐receptor interaction are crucial for translating signaling activation at the cellular level to effects of AVP and OT ligands on social behavior. Knowledge of how OT alters neuronal structure and function has the potential to both identify mechanisms that produce social dysfunction and to inform the development of therapeutic agents.
PPARα‐targeted mitochondrial bioenergetics mediate repair of intestinal barriers at the host‐microbe intersection during SIV infection
Authors: Crakes Katti R. , Rocha Clarissa Santos , Grishina Irina , Hirao Lauren A. , Napoli Eleonora , Gaulke Christopher A. , Fenton Anne , Datta Sandipan , Arredondo Juan , Marco Maria L. , Sankaran-Walters Sumathi , Cortopassi Gino , Giulivi Cecilia , Dandekar Satya 
Published: April 2020

Chronic gut inflammatory diseases are associated with disruption of intestinal epithelial barriers and impaired mucosal immunity. Human immunodeficiency virus‐1 (HIV) causes depletion of mucosal CD4+ T cells early in infection and disruption of gut epithelium resulting in chronic inflammation and immunodeficiency. Although anti‐retroviral therapy (ART) is effective in suppressing viral replication, it is incapable of restoring the ‘leaky gut’ which poses an impediment for HIV cure efforts. Strategies are needed for rapid repair of the epithelium to protect intestinal microenvironments and immunity in inflamed gut. Using an in vivo non‐human primate intestinal loop model of HIV/AIDS, we identified the pathogenic mechanism underlying sustained disruption of gut epithelium and explored rapid repair of gut epithelium at the intersection of microbial metabolism. Molecular, immunological, and metabolomic analyses revealed marked loss of peroxisomal proliferator‐activated receptor alpha (PPARα) signaling, predominant impairment of mitochondrial function and epithelial disruption both in vivo and in vitro. To elucidate pathways regulating intestinal epithelial integrity, we introduced probiotic Lactobacillus plantarum into Simian immunodeficiency virus (SIV)‐inflamed intestinal lumen. Rapid recovery of the epithelium occurred within 5 hours of L. plantarum administration and independent of mucosal CD4+ T cell recovery and in the absence of ART. This intestinal barrier repair was driven by L. plantarum‐induced PPARα activation and restoration of mitochondrial structure and fatty acid beta‐oxidation. Our findings elucidate the critical role of PPARα 1) at the intersection between microbial metabolism and epithelial repair in virally‐inflamed gut and 2) as a potential mitochondrial target for restoring gut barriers in other infectious or gut inflammatory diseases. These data highlight an opportunity to investigate the evolutionary synergism between host mitochondria and gut microbes to restore mucosal health in lieu of full immune recovery.Support or Funding InformationThis study was supported by the National Institutes of Health grant R01 AI123105. KRC received a pre‐doctoral training grant F30 AI150462.