bluuebunny/crossref_metadata_2025 huggingface dataset:
Keystone Species in the Pathogenic Process of NAFLD
Authors: Fang Sa , Wu Dingfeng , Yang Zihuan , Jiao Na , Lin Yunzheng , Li Xiaoyi , Xiao Jing , Yuan Meifei , Zhu Ruixin , Zhu Lixin
Published: June 2021

Background

The role of gut microbiome in the pathogenic process of NAFLD has been increasingly recognized. However, the keystone species in the gut of NAFLD, the critical factor(s) in maintaining the stability of the entire gut microbiome, has not been reported.

Method

SparCC method was used to construct the microbial interaction ecosystem of human gut microbiome in healthy condition. PageRank algorithm was used to evaluate the importance of each bacterial species in healthy gut micro‐ecology system. Subsequently, the keystone species required for maintaining the stability of gut micro‐ecological system was identified on the basis of network stability assessment. Finally, the keystone species specific for NAFLD was determined by the quantitative analysis of gut microbiome between NAFLD patients and healthy people. And the relevant OTUs were annotated at species‐level based on phylogenetic trees. Furthermore, the same keystone species identification analysis was also conducted in NAFLD rats induced by high‐fat diet.

Results

Here we reconstructed the microbial interaction ecosystem of healthy human gut microbiome, consisting of 61 bacterial species and 90 pairwise interactions. Wherein, Bacteroides ovatus, Faecalibacterium prausnitzii, Barnesiella intestinihominis, Odoribacter splanchnicus, Roseburia faecis, Eubacterium ventriosum, Clostridium leptum were the seven keystone species that maintained the stability of the gut micro‐ecological system. The results of quantitative analysis indicated that the abundance of B. intestinihominis, E. ventriosum and C. leptum decreased significantly in the gut of NAFLD patients, and they were identified as the keystone species specific for NAFLD. These species could participate in the initiation and development of NAFLD by regulating the metabolic process of bile acid signaling and the activity of inflammasome. In addition, similar results were observed in the gut microbiome of NAFLD rats.

Conclusion

This research identifies the keystone species specific for NAFLD, and highlights a novel target for the precise prevention and treatment of NAFLD based on gut microbiome. Importantly, our strategy for the identification of the keystone species can be extended to other ecological systems.

Support or Funding Information

This work was supported by National Natural Science Foundation of China 81770571 (to LZ), 81774152(to RZ), 41530105 (to RZ).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rapastinel as a non‐opioid treatment for opioid dependence
Authors: Ferrante Julia Renee , Lamichhane Nidesh , Kuhn Cynthia , Armstrong Christopher , Walker David , Patkar Ashwin
Published: June 2021

An increasing number of opioid overdose deaths have resulted from the rising availability of both prescription and nonprescription opioids. Pharmacotherapies, such as buprenorphine and methadone, are used to treat those dependent on opioids. However, these commonly used pharmacotherapies are opioid partial agonists and agonists, so the patient remains in an opioid‐dependent state throughout treatment. Treatment requires long‐term tapering with buprenorphine or methadone, during which withdrawal symptoms can occur. These limitations show a clear need for a non‐opioid pharmacotherapy. Drugs that target the NMDA receptor have been proposed as alternatives to opioid‐targeted pharmacotherapies. The NMDA receptor is involved in the maintenance of opioid dependence and its blockade may reverse the neuroadaptive changes induced by opioid dependence. Regulation of the NMDA receptor could accelerate treatment of opioid dependency (Glass, 2011). Ketamine, an NMDA antagonist, has potential as a treatment, but has associated side effects and potential for abuse. Rapastinel is a novel drug marketed as an antidepressant, and it acts as a partial agonist of the NMDA receptor complex. The negative side effects reported with ketamine have not been reported with rapastinel (Moskal et al., 2016). Since rapastinel acts as a partial agonist and has no reported side effects, it may be a more tolerable treatment option for those dependent on opioids. Male and female adolescent (between 28 and 30 days old) Sprague‐Dawley rats were injected with morphine in increasing doses twice a day for five days (5 mg/kg/day–25 mg/kg/day) to induce opioid dependency. On day six, rats were injected with naloxone (1 mg/kg) and withdrawal signs were quantified. Rats were then given either ketamine injections (1 mg/kg) twice daily (n=12), rapastinel injections (5 mg/kg) every other day (n=14), or saline injections (n=24). On day nine, rats were again administered naloxone (1 mg/kg), and withdrawal signs were quantified to measure efficacy of ketamine and rapastinel. Rapastinel treated rats exhibited significantly fewer withdrawal signs than those treated with ketamine, which did not differ from controls. These results show that rapastinel significantly enhances recovery from opioid dependence.

Support or Funding Information

Funding provided by ASPET, Duke University SURPH program, and Duke University Department of Pharmacology and Cancer Biology

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A novel conformation‐sensitive antibody‐based strategy to explore PKC signaling induced by opiate activation of μ‐opioid receptors
Authors: Duarte Mariana Lemos , Devi Lakshmi
Published: June 2021

Opiates such as morphine and fentanyl are the most widely used analgesics for the treatment of pain; however, prolonged treatment with opiates decreases the responsiveness of opioid receptors, leading to the development of tolerance and opiate addiction. Recent alarming reports of the widespread misuse of fentanyl leading to the ‘Opioid Epidemic” have begun to refocus scientific investigations into the mechanisms of action of opioid receptors. Despite intense research efforts, the molecular mechanisms underlying opioid addiction remain unknown contributing to a public health challenge. Activation of opioid receptors leads to the initiation of a number of signaling cascades, resulting in enhanced protein phosphorylation. Among the various kinases, protein kinase C (PKC) has been reported to play a crucial role in the desensitization of opioid receptors. To date, the majority of studies with PKC have used genetic and/or pharmacological manipulation of PKC to explore the dynamics of PKC activation. In order to investigate the activation of endogenous PKC in native tissue, we developed conformational‐sensitive antibodies that are able to detect time‐ and ligand‐mediated activation of native PKC. We also generated antibodies to several substrates of PKC including pPP2A (regulatory subunit phosphoS90 PPP2R5D), phosphoSer143 MARCKS, and phosphoS375/T376 MOR. We used a high‐throughput microscopy approach to explore the temporal dynamics of opiate‐mediated PKC signaling and track the temporal dynamics of PKC activation in cells treated with PMA, morphine or fentanyl. Interestingly, in contrast to the PKC activation induced by morphine that is rapid but transient the activation by fentanyl is slow and sustained for a longer period of time. We are able to detect rapid activation of PKC in the striatum of animals acutely treated with morphine suggesting that the conformation‐sensitive activation, state sensitive anti‐PKC antibodies serve as hitherto unavailable, critical tools to investigate the role of PKC in opioid receptor signaling in particular and PKC‐mediated signaling in general.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Icariside II, a phosphodiesterase 5 inhibitor, attenuates brain injury in focal cerebral ischemia/reperfusion rats: Involvement of inhibition of GSK‐3β‐mediated activation of autophagy
Authors: Gao Jianmei , Li Fei , Long Long , Xu Fan , Feng Linying , Liu Yuangui , Shi Jingshan , Gong Qihai
Published: June 2021

Cerebral ischemia/reperfusion (I/R) is always exacerbated neuronal damage when timely restoring cerebral blood flow after ischemia stroke. Cerebral I/R results in excessive autophagy and leads to neuronal demise. The present study was designed to investigate the role of autophagy during ICS II‐mediated neuroprotection on cerebral I/R in vivo and in vitro. Here, we found that ICS II not only protected against middle cerebral artery occlusion‐induced injury in rats, but also attenuated primary cortical neurons injury upon oxygen‐glucose deprivation. Moreover, autophagy was activated as evidenced by triggered autophagic influx and increased ATG expressions after stimulation of cerebral I/R. However, ICS II signifcantly inhibited excessive autophagy as evidenced by repressing autophagic flux, combined with decreasing ATG expressions including LC3 II, Beclin 1, ATG5 and ATG7. Interestingly, ICS II, a PDE5 inhibitor, not only restored cGMP/PKG pathway, but also inactivated GSK‐3β through mediating phosphorylation at tyr216 and ser9. Most intrugingly, siGSK‐3β obviously promoted the beneficial effect of ICS II, which further confirmed that the neuroprotective mechanism of ICS II is, at least in partly, through inactivating GSK‐3β. Furthermore, we reveled that the cGMP/PKG pathway acts as the upstream of GSK‐3β inactivation in the inhibitory effect of ICS II on cerebral I/R‐induced autophagic neuronal death, as evidenced by GSK‐3β inhibitor or PKG inhibitor in vitro. Most importantly, the findings demonstrated that ICS II protected against cerebral I/R‐induced injury via mediating PKG/GSK‐3β/autophagy axis. This study will raise the possibility that ICS II may be exploited into a potent PDE 5 inhibitor against ischemia stroke.

Support or Funding Information

This work was supported by Natural Science Foundation of China Natural Science Foundation of China (Grant No. 81560666), Program for excellent young talents of Zunyi Medical University (Grant No. 15zy‐002), Science and Technology Innovation Talent Team of Guizhou Province (Grant No.20154023), The hundred level of high‐level innovative talents in Guizhou Province (Grant No. QKHRCPT 20165684), and Program for Changjiang Scholars and Innovative Research Team in University, China (Grant No. IRT_17R113).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulatory Effect of a Standardised Andrographis Paniculata Aqueous Extract on Urine Metabolomics in a Rat Model of Neuroinflammation
Authors: Sani Dahiru , Kirby Brian P , Basri Hamidon , Stanslas Johnson
Published: June 2021

In this study, the effect of pre‐treatment with standardised andrographis paniculata aqueous extract (APAE) on rats urine metabolomic changes following Lipopolysaccharide (LPS)‐induced neuroinflammation was evaluated based on proton‐nuclear magnetic resonance (1H‐NMR) spectroscopy. Both principal component analysis and orthogonal Partial Least Square‐Discriminant Analysis (OPLS‐DA) revealed that LPS (1 mg/kg) induces metabolic alterations. LPS group showed increased urinary levels of oxoglutarate, creatinine, citrate, allantoin, lactate and hippurate. However, rats pre‐treatment with APAE for 7 days prior to LPS administration prevented the upregulation of these molecules similar to the control group to confer neuroprotection. Our findings imply that metabolomic procedures based on NMR could be applicable to elucidate neuro‐inflammation via the biomarkers exploration. In addition, we proposed a mechanistic metabolic pathway for APAE suppression of neuroinflammation.

Support or Funding Information

This study was supported by the Malaysia Ministry of Agriculture (NRGS grant, NH612D009) and by Government of Sokoto State, Nigeria

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Specific TBC Domain‐Containing Proteins Control the ER‐Golgi‐Plasma Membrane Trafficking of GPCRs
Authors: Wei Zhe , Huang Wei , Wu Guangyu
Published: June 2021

G protein‐coupled receptors (GPCRs) constitute the largest superfamily of cell‐surface signaling proteins. However, the molecular mechanisms underlying their cell surface delivery after synthesis remain poorly understood. Here we screen the TBC domain‐containing proteins, putative Rab GTPase‐activating proteins (GAPs), in the intracellular trafficking of GPCRs and identify several TBC proteins that activity‐dependently regulate the anterograde transport, en route from the endoplasmic reticulum to the Golgi or from the Golgi to the cell surface, of several prototypic GPCR members without affecting other plasma membrane proteins. We also show that TBC1D6 functions as a GAP for Rab26, physically associates with Rab26, and attenuates Rab26 interaction with GPCRs. Furthermore, both overexpression and depletion of TBC1D6 inhibit the post‐Golgi traffic of GPCRs. These data demonstrate for the first time the important roles of the TBC proteins in forward trafficking of nascent GPCRs and reveal novel regulatory mechanisms of GPCR targeting to the functional destination.

Support or Funding Information

NIH R01GM118915

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Therapeutic Assessment and Neuroprotective Effects of a Novel G Protein‐Coupled Estrogen Receptor (GPR30/GPER) Agonist
Authors: Sondergard Chelsea , Gunn Joseph , Pemberton Kyle , Arnatt Christopher K
Published: June 2021

The G protein‐coupled estrogen receptor (GPER) has been implicated in the neuroprotective effects of 17β‐estradiol and selective GPER agonist, G‐1. Currently, G‐1 remains the standard for GPER agonists, necessitating the design and identification of novel candidates. Previously, our group synthesized and biologically evaluated a series of GPER antagonists. Modification of an aromatic moiety on the scaffold altered the pharmacological profile from antagonists to agonists. Using a fluorescence‐based binding assay for the classical estrogen receptors, ERa and ERb, we discovered off‐target binding. Due to the similarity in functionalization to estradiol, we hypothesized that the hydrogen bonding abilities of the initial candidate were responsible for the observed off‐target binding. To mitigate off‐target binding, modifications were made to reduce the hydrogen bonding ability. For each resulting candidate, the EC50 value was determined through in vitro biochemical screening. From these studies, we identified a lead compound that exhibited an EC50 value below that of our experimentally determined EC50 value of G‐1. Using E18 rat embryonic hippocampal neurons, the lead compound was shown to increase synaptogenesis. These results suggest that the identified lead compound for GPER may be a potential candidate for neurodegenerative diseases.

Support or Funding Information

Saint Louis University

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Marine Pharmacology and Pharmaceuticals Pipeline in 2018
Authors: Abadines Ivan B , Le Kelly , Newman David J , Glaser Keith B , Mayer Alejandro M
Published: June 2021

The ocean's organisms continue to deliver a chemical biodiversity that contributes to both the global preclinical and clinical pharmaceutical pipelines. Thus, in November 2018, the clinical marine pharmaceutical pipeline consisted of seven marine‐derived drugs approved by the United States Food and Drug Administration (FDA): for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969); for Herpes Simplex Virus, vidarabine (Vira‐A®, FDA‐approved 1976); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia, omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004); for cancer, eribulin mesylate (Halaven®, FDA‐approved 2010), brentuximab vedotin (Adcetris®, FDA‐approved 2011), and trabectedin (Yondelis®, FDA‐approved 2015). The website www.clinicaltrials.gov was searched to determine marine‐derived pharmaceuticals that were in Phase I, Phase II and Phase III of clinical development in November 2018. Confirmation of our findings in the www.clinicaltrials.gov website was conducted by visiting the websites of all pharmaceutical companies supporting the development of compounds currently in Phase I, Phase II and Phase III of clinical development. Our clinical research determined that there were 25 marine‐derived compounds in active clinical trials in November 2018: 6 marine‐derived compounds were in Phase III, 10 compounds in Phase II, and at least 9 compounds in Phase I, many of them auristatin‐containing antibody drug conjugates or ADCs. Regular updates on the clinical marine pharmaceutical pipeline will be posted on the dedicated website: http://marinepharmacology.midwestern.edu/clinical_pipeline.html. Furthermore, it is important to note that the global preclinical marine pharmacology pipeline continues to generate considerable data on multiple pharmacological classes, as highlighted in a recent review: A. M. S. Mayer, A. D. Rodríguez, O. Taglialatela‐Scafati and N. Fusetani. Marine Pharmacology in 2012–2013: Marine Compounds with Antibacterial, Antidiabetic, Antifungal, Anti‐Inflammatory, Antiprotozoal, Antituberculosis, and Antiviral Activities; Affecting the Immune and Nervous Systems, and other Miscellaneous Mechanisms of Action. Marine Drugs 15 (9): 273, 2017. We thus conclude that in November 2018, both the marine pharmacology preclinical and clinical pharmaceutical pipelines continued to remain very active.

Support or Funding Information

Support by the College of Pharmacy, Vicki Sears, Pharmacology Department, College of Graduate Studies and Elizabeth Tallman, Information Technology Services, Midwestern University are gratefully acknowledged.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In vitro screening of Glyphae brevis (Tiliaceae) for anticholinesterases and antioxidant potentials as a ethnomedicinal component of memory enhancing agent in Niger Delta Nigeria
Authors: NWIDU LUCKY LEGBOSI , ELMORSY EKRAMY , CARTER WAYNE GRANT
Published: June 2021

Background

This study investigates the anticholinesterases and antioxidant potential of Glyphae brevis used in ethnomedicine as memory enhancing therapy.

Methods

Glyphae brevis crude methanolic leaf extract (GBCMLE), stembark (GBCMSE) and root (GBCMRE), and fractions: ethyl acetate (GB EAF), butanol (GB BUF), aqueous (GB AQF) were investigated for acetylcholinesterase (AChE) inhibitory activity using Ellman's assay and DPPH free radical scavenging activity. The total Phenolic and Flavonoid contents were also examined and expressed in mg Gallic acid (GAE/g) and quercetin (QUER/g) respectively.

Results

The samples exhibited concentration dependent AChE inhibitions, the leaf: GBCMLE, 52.6%, (IC50 127 μg/mL), GBLEAF, 46.3% (IC50 245 μg/mL), GBL BUF, 24.5% (IC50 425 μg/mL) and GBL AQF, 20.3% (IC50 502 μg/mL); the stem: GBCMSE 68.3% (IC50 19 μg/mL), GBS EAF 172.2% (IC50 172 μg/mL), GBS BUF 65.4% (IC50 21 μg/mL), GBS AQF 85.% (IC50 12 μg/mL); the root: GBCMRE 42.2% (IC50 497 μg/mL), GBR EAF 70.1% (IC50 509 μg/mL), GBR BUF 81.3% (IC50 287 μg/mL) and GBR AQF 60.5% (IC50 521 μg/mL) at a concentration of 200 μg/mL. At this dose, the AChE activities ~20.3 – 52.6%, ~52.2 – 85.6% and ~42.2 – 81.3% for leaf, stem and root respectively of the value observed using eserine 0.02μg/mL. The descending AChE activity is stem > root > leaf.

In the DPPH free radicals scavenging assay, highest activity were demonstrated by GBC MLE and GBL EAF, 72.5 ± 3.1 and 65.2 ± 3.2%; for the stem, GBS AQF and GBS BUF, 80.5 ± 3.4 % and 68.3 ± 3.4 %; and the root, GBR BUF and GBR EAF, 78.2 ±3.3% and 67.0 ±3.1% inhibitions of free radicals respectively at 1 mg/mL concentration. The corresponding IC50 were 327, 473, 278, 327, 211 and 473 μg/mL respectively. Similarly, the TPC and TFC are 88.85 and 31.61, 52.36 and 19.45, 75.67 and 27.22, 55.34 and 24.67, 97.0 and 34.33, 51.87 and 19.29 mg Gallic acid GAE/g and quercetin QUER/g respectively. BUF has the highest correlation (r) values, which means that the contents of BUF extracts are the most important for the antioxidant and AChE properties.

Conclusion

These results indicate that G. brevis is enriched with phenolic and flavonoid contents that exhibit marked anticholinesterase and antioxidant effects..

Support or Funding Information

University of Nottingham Visiting Fellowship

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Perfluorononanoic Acid (PFNA) Suppresses Autophagy and Activates Nrf2 Signaling
Authors: Le Yuan , Cheng Xingguo
Published: June 2021

Perfluorinated compounds (PFCs), a group of persistent environmental pollutants, are frequently detected in human blood. Epidemiological and animal studies show that there are positive associations between higher blood levels of PFCs and greater risk of liver disorders, thyroid disorders, reproductive toxicity, and immunotoxicity. Nuclear factor erythroid 2‐related factor (Nrf) 2, a transcription factor that can be activated by oxidative stress, electrophiles, and accumulated p62 protein after autophagy repression, plays important anti‐inflammatory, anti‐oxidative and cytoprotective roles. We previously reported that PFCs increased the expressions of several Nrf2 target genes in mouse liver. However, the underlying mechanism by which PFCs activate Nrf2 signaling remains largely unknown. The present study was designed to determine whether PFNA activates Nrf2 signaling; and if so, the underlying mechanism. PFNA increased mRNA expressions of Nrf2‐target genes, including Heme oxygenase (Ho/HO) 1 and NAD(P)H: quinone oxidoreductase (Nqo/NQO) 1, as well as nuclear levels of Nrf2/NRF2 protein in both mouse liver and cultured human Hep3B hepatoma cells. To our surprise, PFNA did not produce apparent oxidative stress in both mouse liver and cultured human Hep3B hepatoma cells. We next determined the impact of PFNA on autophagy progression. In both mouse liver and cultured human Hep3B hepatoma cells, PFNA decreased the conversion of microtubule‐associated protein 1 light chain 3 beta (LC3B)‐I into LC3B‐II, and increased cytosolic levels of p62 protein, indicating that PFNA repressed autophagic flux. In addition, PFNA decreased nuclear levels of transcription factor EB (TFEB) protein, an upstream signaling molecule of autophagy. Morphologically, PFNA caused liver injury in both wide‐type and Nrf2‐null mouse liver, evidenced by hepatocyte necrosis and hemorrhage. In consistency with histology study, PFNA (0.1mmole/kg) increased serum AST and ALT levels in both wide‐type mice and Nrf2‐null mice. In addition, PFNA produced clear inflammation foci in Nrf2‐null mouse liver, but not in wide‐type mouse liver. Furthermore, PFNA induced mRNA expressions of pro‐inflammatory cytokines, including IL‐1β, IL‐6, and TNF‐α only in Nrf2‐null mouse liver, but not in wide‐type mouse liver. In conclusion, PFNA activates Nrf2 signaling, primarily mediated by accumulated p62 protein after autophagy repression, which significantly attenuated PFNA‐induced inflammation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phenobarbital Increased Hepatic Toxicity of Acetaminophen due to Cytochrome P450 Induction in Young and Adult Mice
Authors: Shao Xueyan , Wang Pei , Bao Yifan , Chen Liming , Zhong Xiao‐bo
Published: June 2021

Cytochrome P450 (P450) enzymes, phase I drug metabolizing enzymes, are responsible for the metabolism of a variety of substances, including endogenous compounds, xenobiotics and drugs. The expression of P450s could be altered by factors including drug treatment, which may lead to decreased drug therapeutic efficacy or increased toxicity. Phenobarbital is a known inducer of P450s and commonly utilized to treat seizures in children and adults. Although acetaminophen serves as the most commonly used drug for pain and fever in children and adults, an overdose of acetaminophen could lead to severe liver damage. The aim of our study was to examine the short‐term effect of phenobarbital administration on the hepatic toxicity of acetaminophen and the correlation of this effect with the alterations in P450s expression in young and adult mice. Mice were pre‐treated with saline or phenobarbital at the age of Day 10 or Day 59. 24 hours later, saline, 200 mg/kg, or 400 mg/kg acetaminophen was administered to mice. Real‐time polymerase chain reaction (RT‐PCR) was used to detect mRNA expression of P450s. Moreover, liver toxicity was evaluated based on plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Phenobarbital pre‐treatment enhanced ALT and AST levels in response to acetaminophen treatment in young and adult mice. For young mice with phenobarbital pre‐treatment, CYP1A2, CYP2B10 and CYP2C29 mRNA levels were significantly increased in male mice and the mRNA expression of CYP2B10, CYP2C29, CYP2E1 and CYP3A11 showed significant enhancement in female mice. As for adult mice with phenobarbital pre‐treatment, the mRNA expression of CYP2B10, CYP2C29 and CYP3A11 were found to go up in male and female mice, while only male mice showed a significant increase in CYP1A2 mRNA expression. In summary, our data indicate that phenobarbital pre‐treatment elevated hepatic toxicity of acetaminophen as a result of P450s induction in both young and adult mice. Therefore, much attention should be paid to the enhanced hepatic toxicity of acetaminophen when combined with phenobarbital in clinical setting.

Support or Funding Information

This study was supported by the National Institutes of Health National Institute of General Medical Sciences [Grant R01GM‐118367] (to X.‐b.Z.).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In vitro Transporter Screens to Investigate Uremic Solute‐Drugs Interaction
Authors: Chang Shih‐Yu , Chapron Alenka , Imaoka Tomoki Imaoka , Yeung Catherine K
Published: June 2021

Renal tubule secretion via transporter proteins plays a critical role in the renal clearance of drugs and endogenous compounds. We hypothesize that competition between accumulating uremic solutes and drugs for proximal tubule transport results in unpredictable drug exposure in people with kidney disease. The goal of this study was to characterize the potential of prominent uremic solutes, including hippuric acid (HA), indoxyl sulfate (IS), p‐cresol sulfate (pCS) and trimethylamine N‐oxide (TMAO), to inhibit renal tubule transport. We used penciclovir (PEN), oseltamivir carboxylate (OST) and tenofovir (TEN) in our study as model transporter substrates. We conducted systemic screens of transporter inhibition using in vitro assays, including uptake (overexpressed cell lines) and efflux (in‐side out vesicle membranes) transporters, to investigate whether uremic solutes and model drugs are potential inhibitors/substrates of renal uptake and efflux transporters involved in tubular secretion (OAT1/2/3, MRP2/4, MATE1/2K, P‐gp, BCRP). This systematic study of uremic solute ‐ drug interaction demonstrated that some uremic solutes and drugs (PEN and TEN) might compete and inhibit uptake transporters via OATs and efflux via MRPs transporters. A clearer understanding of the interactions between drugs and uremic solutes may lead to personalized approaches for the selection and dosing of medications in people with kidney disease.

Support or Funding Information

5 R01 GM 121354‐02 UREMIA TRANSPORT

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hepatic GSTZ1 Expression in Pregnant Ewes and Their Offspring: Influence of Treatment with Dichloroacetate
Authors: James Margaret O , Luzi Michelle , Miller Megan L , Faux Laura Rowland , Joseph Serene , Stacpoole Peter W , Keller‐Wood Maureen
Published: June 2021

Dichloroacetate (DCA) is an investigational drug that can stimulate mitochondrial energy metabolism by inhibiting pyruvate dehydrogenase kinase. We hypothesize that DCA could be used in high‐risk pregnancy to reduce perinatal mitochondrial deficiency and resulting cardiac dysfunction in at‐risk fetuses and are testing this postulate in a pregnant sheep model. DCA is metabolized by the enzyme glutathione transferase zeta1 (GSTZ1) to glyoxylate. GSTZ1 is not expressed in human fetal liver, but expression rises after birth. Furthermore, DCA inactivates GSTZ1, resulting in altered pharmacokinetics with repeated dosing. The objectives of this research were (1) to investigate the relative expression of GSTZ1 in maternal and fetal sheep liver from untreated controls close to full term (140 days gestation) and (2) to determine if treatment of sheep with DCA resulted in reduced expression of GSTZ1. We administered DCA intravenously to either the ewe or the fetus at daily divided doses of 25 mg/kg for three to five days prior to sacrifice 24 h after the last dose. We used samples of maternal and fetal liver to prepare cytosol and mitochondria and employed a custom‐made polyclonal antibody to rat GSTZ1 to measure expression relative to a single rat hepatic cytosol standard by Western blot. The sheep liver samples exhibited good cross‐reactivity to the rat antibody. Surprisingly, expression of GSTZ1 in the full‐term fetal liver cytosol fractions from controls was similar to that in the control maternal liver. Treatment with DCA resulted in reduced expression in both maternal and fetal liver cytosol fractions to less than 10% of the control values. In control fetuses, mitochondrial expression of GSTZ1 was 17.6 ± 7.6% of the cytosolic expression. In the DCA‐treated group, mitochondrial expression decreased and levels were undetectable in one‐half of the fetal sheep. These studies demonstrate that (1) GSTZ1 expression in liver cytosol and mitochondria in the late fetal period varies with animal species and (2) repeated treatment with DCA results in reduced GSTZ1 expression, which is likely to alter DCA pharmacokinetics.

Support or Funding Information

Supported in part by the US Public Health Service, grant R21HD91599

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Angiotensin II Inhibits DDAH1‐nNOS Signaling through AT1R and μOR Dimerization in the Central Nervous System to Modulate Central Control of Blood Pressure
Authors: Cheng Pei‐Wen , Sun Gwo‐Ching , Tseng Ching‐Jiunn
Published: June 2021

Aims

G protein‐coupled receptors (GPCRs) play an important role in drug therapy, and represent one of the largest families of drug targets. Blockade of angiotensin II type 1 receptor (AT1R) signaling has been shown to alleviate hypertension and improve outcomes in patients with heart failure. GPCRs formations or desensitization and trafficking can lead to pathophysiological processes. The aim of the present study was to investigate that may Ang II through induce AT1R and μOR dimerization in the NTS leads to the progress of hypertension.

Method

Formation of heterodimers was assessed with proximity ligation assays. Microinjections of μOR or α2A‐AR agonists into the NTS and measured changes in BP of WKY rats. Real‐time PCR and immunoblotting analyses were used to quantify RNA and protein expression levels.

Result

The levels of μOR and α2A‐AR heterodimer were increased and the extracellular signal–regulated kinases 1/2T202/Y204, ribosomal protein S6 kinaseT359/S363, nNOSS1416 phosphorylation; DDAH1expression were abolished in the NTS of adult spontaneously hypertensive rats (SHRs), however, the overexpressed endomorphins‐2 in the NTS of SHR when compared with young SHR. Microinjection of μOR agonist into NTS induced persist presser effect of WKY rats, decreased NO production and DDAH1. Immunoblotting analysis showed that μOR agonist significantly reduced the DDAH1 activity and decreased the expression of nNOS phosphorylation. Additionally, Ang II inhibitors (losartan) significantly decreased the BP and abolished AT1R‐induced both formation of AT1R and μOR heterodimers and α2A‐AR and μOR heterodimers. Besides, also has the same founding, losartan significantly enhanced nNOSS1416 phosphorylation and DDAH1 expression. In summary, these results represent a novel finding that Ang‐II induces endomorphins‐2 and abolished DDAH1 through enhancing the formation of AT1R and μOR heterodimers formation in the NTS to progress hypertension.

Support or Funding Information

This work was supported by funding from the Ministry of Science and Technology (MOST106‐2314‐B‐037‐022‐MY2, MOST107‐2320‐B‐075B‐001) and Kaohsiung Veterans General Hospital (VGHKS107‐175) (to P.‐W. C). The authors declare that they have no competing interests.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The role of Sirt3‐AMPK in mediating oxidative stress‐induced hypertension in the brain stem nuclei of rats
Authors: Ho Chiu‐Yi , Cheng Pei‐Wen , Chen Hsin‐Hung , Tseng Ching‐Jiunn
Published: June 2021

Essential hypertension is the most common cardiovascular disease and is a major risk factor for stroke and heart disease. It is known the nucleus tractus solitarii (NTS) located on the dorsal medulla play an important role in regulating cardiovascular responses. More recently, metabolic syndrome‐induced phosphorylated AMPK defect has been demonstrated to underline the initiation of superoxide elevation. In recent years, fructose induces a decrease in SOD2 activity, which leads to a large increase in superoxide and a decrease in the pathway of Akt‐nNOS. Studies have found that stimulation with abnormal diet factor can promote the reduction of Sirt3 and increase the content of superoxide. Therefore, we hypothesize that the SOD2 activity in the NTS was inhibited via Sirt3‐AMPK dysfunction induced superoxide expression and hypertension. During this study, the rats were fed 10% fructose with or without natural compound 3H‐1,2‐Dithiole‐3‐thione (D3T) for 4 weeks with the D3T treatment beginning at week 2. The systolic blood pressure and serum fasting high‐density lipoprotein decreased significantly in the D3T‐treated group compared fructose‐fed group after feeding D3T for 2 weeks. The protein levels of Sirt3, and AMPKT172 showed a significant increase in the D3T‐treated group compared fructose group after feeding D3T for 2 weeks by immunoblotting analyses. However, the acetylation of SOD2 were significantly decreased in the D3T‐fed group compared the fructose‐fed groups. Superoxide anion expression levels in the NTS were clearly eliminated in the D3T‐fed group compared the fructose‐fed groups. Collectively, fructose induced Sirt3 and AMPK defect and acetylation of SOD2 in the NTS of fructose‐fed rats, while treatment of D3T in the fructose‐fed rat reversed the effect. This study suggest that fructose‐induced hypertension may through Sirt3‐AMPK to mediate the acetylation of SOD2 and causes superoxide generation in the central nervous system. Our findings suggested a better understanding of complex cardiovascular neural pathways and may provide a new link between hypertension and neuronal superoxide that could be targeted for the treatment of cardiometabolic diseases.

Support or Funding Information

This work was supported by funding from the Ministry of Science and Technology (MOST‐107‐2320‐B‐075B‐002).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Type 2 Diabetic Human Coronary Vascular Smooth Muscle Cells Are Less stiff and Less Adhesive to Fibronectin
Authors: Trask Aaron J. , McCallinhart Patricia E.
Published: June 2021

Previous data from our laboratory has demonstrated that type 2 diabetic (T2DM) coronary resistance microvessels (CRMs) undergo inward hypertrophic remodeling associated with reduced stiffness and reduced coronary blood flow in both mice and pig models. Further analysis revealed that the reduced stiffness was in part due to increased tissue elastin expression and decreased vascular smooth muscle cell (VSMC) stiffness in mice. The goal of the present study was to test the hypothesis that T2DM coronary VSMCs from humans are less stiff than normal. De‐identified, low‐passage normal and T2DM human coronary VSMCs were purchased from Lonza and ATCC for all experiments (n=4–5 per group). Using an atomic force microscope with probes coated with fibronectin (FN), a nano‐indentation protocol was used to measure cellular stiffness and adhesion to FN. Elastic modulus, a measurement of cellular stiffness, was reduced in T2DM human coronary VSMCs compared to normal (Normal: 3.60 ± 0.32 kPa vs. T2DM: 2.75 ± 0.22 kPa, p<0.05). Additionally, T2DM human coronary VSMCs were less adhesive to fibronectin compared to normal human coronary VSMCs (Normal: 58.3 ± 7.7 vs. T2DM: 43.5 ± 4.9 pN, p=0.17). Immunoblot analysis showed that T2DM human coronary VSMC expressed reduced vinculin (Normal: 1.0 ± 0.17 vs. T2DM: 0.31 ± 0.11, p<0.05) and increased cofilin (Normal: 1.0 ± 0.16 vs. T2DM: 1.8 ± 0.35, p=0.052) compared to normal human coronary VSMCs; this expression profile favors a less stiff cell phenotype given that vinculin deficient mice exhibit reduced cytoskeletal stiffness and that increased cofilin binding to F‐actin reduces stiffness. These data show that reductions in stiffness and adhesion to the ECM of diabetic CRM VSMCs may be due to underlying alterations in the cytoskeletal arrangement. Collectively, reduced stiffness in T2DM human coronary VSMCs may drive whole tissue microvascular mechanical properties to account for reduced CRM stiffness. These findings will inform future studies on the mechanisms of stiffness in the diabetic coronary circulation.

Support or Funding Information

NIH R00 HL116769, NIH R21 EB026518, and Nationwide Children's Hospital to AJT

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Estrogen Receptor‐α Counterbalances the Endotoxic Inflammatory Response and Associated Arterial Baroreflex Dysfunction in Ovariectomized Rats
Authors: El‐Lakany Mohammed A , Fouda Mohamed A , El‐Gowelli Hanan M , El‐Mas Mahmoud M
Published: June 2021

Evidence from clinical and experimental studies suggests that the female population is tolerant to inflammatory and cardiovascular anomalies of endotoxemia. In this study we investigated the possibilities that female rats are also protected against the arterial baroreceptor depressant effect of endotoxemia and that this interaction is modulated by gonadal hormones. The effect of i.v. lipopolysaccharides (LPS, 10 mg/kg) on baroreflex chronotropic activity were assessed in conscious sham‐operated (SO) female rats as well as in ovariectomized (OVX) rats supplemented with or without gonadal hormones. Baroreflex curves relating changes in chronotropic responses to increases or decreases in blood pressure evoked by i.v. doses (1–16 μg/kg) of phenylephrine (PE) and sodium nitroprusside (SNP), respectively, were constructed and slopes of the curves were taken as measures of baroreflex sensitivity (BRSPE, BRSSNP). While LPS had no effect on baroreflex activity tested by PE in SO rats, it caused downward shifts in baroreflex curves generated by SNP and significant reductions (by ~ 25%) in slopes of the curves (BRSSNP). Compared with SO rats, significantly greater reductions in BRSSNP were caused by LPS in OVX rats (~ 50%), inferring a restraining influence for gonadal hormones on LPS‐BRSSNP interaction. The BRSSNP depressant effect of LPS in OVX rats was largely eliminated after supplementation with PPT (estrogen receptor‐α agonist), in contrast to no effect for DPN (estrogen receptor‐β agonist), estrogen, or medroxyprogesterone. Immunohistochemical studies revealed that PPT was more effective than estrogen or DPN in attenuating the LPS‐evoked increases in myocardial expressions of NF‐κB and iNOS in OVX rats. Further, the three estrogenic drugs caused comparable increases in myocardial HSP70 expression in the same rat model. None of these molecular entities was improved by medroxyprogesterone. These findings preferentially implicate antiinflammatory pathways of estrogen receptor‐α in the alleviation of baroreflex dysfunction induced by endotoxemia in female rats.

Support or Funding Information

Supported by the Science and Technology Development Fund, Egypt (STDF Grant No. 14895)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of hydrogen sulfide on carbohydrate metabolism and blood pressure in obese type‐2 diabetic rats
Authors: Kashfi Khosrow , Gheibi Sevda , Jeddi Sajad , Ghasemi Asghar
Published: June 2021

Aim

Diabetes is associated with altered endogenous hydrogen sulfide levels, which plays an important role in regulating glucose homeostasis. Here we report on the long‐term effects of sodium hydrosulfide (NaSH) administration on carbohydrate metabolism in obese type 2 diabetic rats.

Method

Type‐2 diabetes was established using high fat‐low dose streptozotocin. Rats were treated for 9 weeks with intraperitoneal injections of NaSH (0.28, 0.56, 1.6, 2.8, and 5.6 mg/kg) or vehicle. Serum fasting glucose was measured weekly for one month and then at the end of the study. Fasting serum insulin was measured before and after the treatment. At the end of the study, glucose tolerance, pyruvate tolerance, and insulin secretion were determined and blood pressure was measured.

Results

In diabetic rats, NaSH at 1.6–5.6 mg/kg increased serum glucose and decreased serum insulin; NaSH at 0.28 and 0.56 mg/kg had no effect on these parameters. High doses of NaSH in diabetic rats decreased glucose and pyruvate tolerance as well as insulin secretion; 0.28 mg/kg had no effect, 0.56 mg/kg improved glucose tolerance but not pyruvate tolerance and insulin secretion. Diabetic rats had higher BP than controls; all doses of NaSH decreased blood pressure in these cohorts. In controls, NaSH (5.6 mg/kg) decreased insulin secretion as well as glucose and pyruvate tolerance, while increased serum glucose. NaSH at 1.6–5.6 mg/kg decreased blood pressure in controls.

Conclusion

In diabetic rats high doses of NaSH increased serum glucose level, decreased serum insulin and impaired glucose and pyruvate tolerance, a low dose decreased serum glucose during glucose tolerance. NaSH decreased blood pressure at all doses.

Support or Funding Information

KK: Supported in part by the National Institutes of Health [R24DA018055; R01GM123508]. SG, SJ, AG: Supported in part by Shahid Beheshti University of Medical Sciences[grant No. 1396/D/93065], Tehran, Iran.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Generic Recombinant Factor VIIa is Comparable to the Branded Novoseven in In‐Vitro and Pharmacokinetic Studies in Primates
Authors: Sadeghi Nasir , Kouta Ahmed , Duff Rick , Iqbal Omer , Abro Schuharazad , Hoppensteadt Debra , Jeske Walter , Mehrotra Siddharth , Walenga Jeanine , Iacobelli Massimo , Fareed Jawed
Published: June 2021

Background

Recombinant factor VIIa (NovoSeven®, NovoNordisk, Bagsvaerd, Denmark) is used in the management of bleeding in patients with hemophilia. A generic version of NovoSeven® is also developed for the same indication (AryoSeven™, AryoGen, Tehran, Iran). The purpose of this study was to compare several batches of generic of rFVIIa, namely AryoSeven™, with multiple commercially available batches of branded NovoSeven® to determine biosimilarity.

Methods

Four commercially available random lots of Novoseven® were obtained through US and European sources and compared to 5 clinical batches of AryoSeven™. These drugs were profiled in vitro using surface enhanced laser desorption ionization (SELDI) mass spectrometry, gel electrophoresis (GE), immunoblotting studies, and procoagulant activity as measured by a clot based method. Thrombin generation studies were carried out using NHP supplemented with 0–1.0 ug/ml rFVIIa. The pharmacokinetic profiles of one representative batch each of the NovoSeven® and AryoSeven™ were studied in primates after a 50 ug/kg bolus IV.

Results

The molecular weight profile of all rFVIIa preparations were comparable. The GE and immunoblotting studies of the two groups of agents showed a comparable profile. The two FVIIa products produced comparable procoagulant effects in the PT and aPTT assays. In the plasma prepared from rFVIIa‐supplemented whole blood, the products produced comparable effect on the PT and aPTT assays. In platelet rich plasma (PRP) and platelet poor plasma (PPP) AryoSeven™ and NovoSeven® at concentrations from 0.125 to 1 μg/ml did not produce any measurable change in either the PT or aPTT assay. The degree of reversal of anticoagulation in samples collected from patients treated with oral anticoagulants was similar in terms of relative correction of the plasma INR with both FVIIa preparations over the concentration range of 0.03 to 1 μg/ml. In the thrombin generation studies NovoSeven® and AryoSeven™ produced similar effects. In the primate pharmacokinetic studies at a dose of 50 ug/kg, both groups exhibited similar pharmacokinetic profiles in terms of biologic half‐life, Cmax, volume of distribution and other parameters.

Conclusions

These studies demonstrated bioavailability between the branded and generic products. The pharmacokinetic profile of the NovoSeven® and AryoSeven™ as studied in primates by measuring the factor VIIa level for up to 4 hours post‐administration were also found to be comparable. These results support the hypothesis that NovoSeven® and AryoSeven™ are biosimilar. Further clinical validation studies are warranted.

Support or Funding Information

None.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.