bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of non‐contingent ethanol administration on self‐administration of ethanol in cynomolgus macaques (658.7)
Authors: Allen Daicia , Helms Christa , Grant Kathleen 
Published: June 2021

After chronic ethanol self‐administration animals show decreased variability in daily drinking, suggesting titration of dose. Previous studies found that self‐administration with limited access to ethanol was resistant to experimenter‐administered ethanol, resulting in intake of a greater‐than‐average daily dose. The current study examined the effects of ethanol gavage on daily intake with continuous access. Ten adult male cynomolgus macaques were induced to drink 4% (w/v) ethanol using a schedule‐induction procedure, followed by 22 hours/day continuous access to ethanol and water. Meals were given 3 times per day at 0, 2, and 4 hours after session start. After 6 and 12 months of daily access, animals were placed in a primate chair and administered ethanol (1.0 g/kg and 1.5 g/kg, i.g.) or saline (i.m.) 2 hours before a daily session. After 6 months of ethanol consumption, there was a robust dose‐dependent decrease in drinking after the challenges (p<0.001), indicating that dose was titrated. Interestingly, at the 12‐month time point, drinking behavior was resistant to the pretreatment, and significant differences from saline were only observed in the 1.5g/kg dose (p<0.05). The dose of self‐administered ethanol was not correlated with blood ethanol concentration after i.g. ethanol, measured immediately before the session. These data suggest that extended access to ethanol self‐administration in monkeys results in intakes that are less sensitive to pharmacological feedback.Grant Funding Source: Supported by AA013510 (Grant)
Delta opioid receptor ‐ kappa opioid receptor heteromer signaling in peripheral nociceptors (659.1)
Authors: McGuire Blaine , Sullivan Laura , Chavera Teresa , Clarke William , Berg Kelly 
Published: June 2021

We have found that DOR‐KOR heteromers form in peripheral nociceptors (i.e., pain‐sensing neurons) (Berg et al., 2012, Mol Pharmacol 81:264‐272). Here we sought to better understand DOR‐KOR heteromer function in peripheral nociceptors. We compared the signaling characteristics of the putative DOR‐KOR heteromer selective agonist, 6’‐GNTI, to that of the selective KOR agonist, U50488. In a rat behavioral model of thermal allodynia, intraplantar (i.pl.) injection of U50488 produced a biphasic (inverted “U”) dose response curve (DRC). The descending limb of the DRC was blocked by the ERK inhibitor, U0126 (10μg; i.pl.) suggesting that ERK mediates the downward phase. By contrast, 6’‐GNTI produced a robust, monotonic DRC that was unaffected by U0126. In primary cultures of peripheral nociceptors, U50488‐mediated activation of ERK was pertussis toxin (PTx) sensitive, whereas 6’‐GNTI‐mediated ERK activation was PTx insensitive. Although both U50488 and 6’‐GNTI inhibit adenylyl cyclase (AC) with similar potency and efficacy, U50488‐mediated inhibition of AC desensitized with repeated U50488 exposure, an effect that was blocked by U0126. Interestingly, 6’‐GNTI‐mediated inhibition of AC did not desensitize with repeated 6’‐GNTI exposure. These data suggest that DOR‐KOR heteromers may have unique signaling characteristics in peripheral nociceptors that make them promising therapeutic targets for pain.Grant Funding Source: DA024865 and COSTAR 2T32DE014318‐12A1
Hyperbaric oxygen treatment suppresses withdrawal signs in opioid‐dependent mice (659.12)
Authors: Nicoara Daniel , Zhang Yangmiao , Nelson Jordan , Brewer Abigail , Shirachi Donald , Quock Raymond 
Published: June 2021

HBO2 therapy has been approved by the Food and Drug Administration for limited clinical conditions (Gesell, The Hyperbaric Oxygen Therapy Committee Report: Indications and Results, 12th Ed, 2008). It has been reported that HBO2 treatments reduce opioid withdrawal in human subjects (Epifanova, Anesteziol Reanimatol, May‐June 1995 issue, 34‐39). However, no further research has been conducted to investigate the mechanism of HBO2 in reducing opioid withdrawal. The purpose of this research was to determine whether HBO2 treatment could suppress signs of withdrawal in opioid‐dependent mice. Male NIH Swiss mice received repeated s.c. injections of morphine sulfate or saline twice a day for 4 days. The daily dose of morphine sulfate was increased progressively from 50 mg/kg on day 1 to 125 mg/kg on day 4. On day 5, opioid withdrawal was precipitated by intraperitoneal injection of 5.0 mg/kg naloxone. Withdrawal signs in mice, including jumping, forepaw tremor and wet dog shakes, were recorded for 30 min and scored. Sixty min prior to the naloxone injection, different groups of mice received a 30‐min or 60‐min HBO2 treatment at 3.5 atmospheres absolute. The results show that HBO2 treatment caused a dose‐dependent reduction in withdrawal jumping, forepaw tremor and wet dog shakes in opioid‐dependent mice.Grant Funding Source: Supported by NIH Grant AT‐007222 and the Allen I. White Distinguished Professorship.
Age‐related changes in peripheral nociceptor function (659.2)
Authors: Sullivan Laura , Jamshidi Raehannah , Chavera Teresa , LoCoco Peter , Dahlhausen Christine , Berg Kelly , Clarke William 
Published: June 2021

Effective and safe pain management in the elderly is a major medical challenge. Furthermore, little is known regarding the effect of aging on nociceptor (pain‐sensing neuron) responsiveness or the function of opioid receptors expressed on nociceptors. Here, we compared the effects of the sensitizing agent, bradykinin (BK), and delta and kappa opioid receptor (DOR, KOR) agonists on nociceptors in young (4‐months‐old) and aged (26‐months‐old) Fisher x Brown Norway rats. Behavioral responses to noxious heat, cold, or mechanical stimulation were determined following intraplantar injections of BK. For all three stimuli, aged rats had a greater allodynic response to BK. In primary cultures, activation of phospholipase C in nociceptors from aged rats was more sensitive to BK than those from young rats. In addition, both opioid agonists, SNC80 (DOR) and Salvinorin A (KOR), produced greater antinociceptive effects in aged rats compared to young rats. Similarly, opioid receptor‐mediated inhibition of adenylyl cyclase activity was greater in nociceptors from aged rats. Overall, our results indicate that aging enhances allodynic effects of the inflammatory mediator, BK, as well as increases antinociceptive effects of opioid agonists. We propose that peripherally‐restricted opioids may be a valuable analgesic strategy for treating pain in the elderly.Grant Funding Source: Supported by T32DA031115, Baptist Health Foundation, Nathan Shock Center of Excellence AG013319
Mechanisms of long‐term regulation of peripheral kappa opioid receptor function by the antagonist, norBNI (659.4)
Authors: Jamshidi Raehannah , McGuire Blaine , Sullivan Laura , Chavera Teresa , Clarke William , Berg Kelly 
Published: June 2021

KOR is known to regulate cellular signaling cascades including c‐Jun N‐terminal Kinase (JNK). We have found that norBNI, a prototypical KOR “antagonist”, has “agonist” activity towards JNK, leading to long‐term reduction in some aspects of KOR function in peripheral pain‐sensing neurons. Using a rodent behavioral model of nociception, we have found that KOR‐mediated peripheral antinociception is abolished 2 and 7 days following a single intraplantar injection of norBNI. This effect was blocked by pre‐injection of a selective JNK inhibitor, SP600125. Treatment of pain‐sensing neurons in culture with norBNI for 1h, followed by washing, eliminated KOR‐mediated inhibition of adenylyl cyclase, measured 24h later, which was blocked by SP600125. However, KOR‐mediated activation of extracellular signal‐regulated kinase (ERK) was unaffected by norBNI treatment.Since JNK is a well‐known activator of transcription factors, ultimately leading to protein synthesis, we sought to determine if protein synthesis is required for the long‐term effects of norBNI. Prior treatment with the protein synthesis inhibitor, cycloheximide, eliminated effects of norBNI, suggesting that increased synthesis of an unknown protein leads to prolonged reduction in some KOR‐mediated responses. These data provide strong evidence that norBNI is a protean ligand which has powerful capacity to regulate peripheral KOR function.Grant Funding Source: Supported by: DA024865, T32DA031115, TST128233/151325
Potential use of the nano‐compound fullerenol for the treatment of paraquat‐induced acute lung injury in rats (660.16)
Authors: Aires Rosária , Socarrás Teresa , Ladeira Marina , Guatimosim Silvia , Ladeira Luis , Campos Paula , Cortes Steyner , Lemos Virginia 
Published: June 2021

The present work aimed at investigating the effect of fullerenol in the treatment of paraquat‐induced acute lung injury (ALI). Wistar rats were separated in control, ALI and ALI+fullerenol groups. In rats given paraquat there was increase of lipid peroxidation in lung, neutrophilia, accumulation of neutrophils in lung and bronchoalveolar lavage (BAL), associated with an increase in pulmonary levels of TNF‐α, IL‐1β, IL‐6 and CINC‐1, and serum levels of TNF‐α and IL‐6. Histological analysis showed inflammatory infiltrate, perivascular and intra‐alveolar edema, hemorrhage, vascular congestion and atelectasis. There was elevation of the basal production of nitric oxide (NO) and hyporesponsiveness of the aorta to vasoconstrictors. Treatment with fullerenol normalized lipid peroxidation, neutrophils number on BAL, lung and blood, the level of IL‐1β and CINC‐1 in lung, and TNF‐α and IL‐6 on serum. All histological parameters were normalized. Furthermore, treatment with fullerenol normalized the basal production of NO and vascular function. Finally, fullerenol increased the survival of animals with ALI. In conclusion, fullerenol combines antioxidative and antiinflammatory activities, reducing pulmonary and vascular alterations, and mortality induced by paraquat. Hence, fullerenol can be an effective and promising drug for the treatment of paraquat‐induced acute lung injury.Grant Funding Source: Supported by FAPEMIG/PRONEX BioNC; CNPq and CAPES
Acute aspiration of graphene sheets evokes transient airway hyperreactivity to methacholine in mice (660.4)
Authors: Fedan Jeffrey , Thompson Janet , Sager Tina , Roberts Jenny , Fedan Jeffrey 
Published: June 2021

Concern exists that the use of graphene sheets (GS) in composite materials might expose manufacturing workers to an inhalation hazard. Studies have shown that GS are cytotoxic in vitro (PC12 cells, fibroblasts) and in vivo in mice (lung granuloma), and Roberts et al. (2013) found non‐oxidized GS with larger lateral dimensions (蠅 5 μm) and a greater number of layers (~20) produced more lung inflammation up to 7 d after aspiration in mice when compared to smaller GS (<1 μm laterally, ~ 4 layers). The lung toxicity of various forms of GS has not been characterized completely. Here, we investigated the effects of GS on basal lung resistance (RL), basal dynamic compliance (CDyn), and reactivity to inhaled methacholine (MCh) aerosol. Mice were given a non‐oxidized GS (5 μm x 5 μm laterally, 7 nm thick equal to ~ 20 layers; 40 µg) suspended in dispersion medium (DM; Porter et al., 2008) or DM (control) via aspiration. RL and CDyn and reactivity to increasing concentrations of MCh aerosol were measured 4 h ‐ 2 mo after GS exposure. Basal RL was increased 4 h post‐exposure but at no other time; basal CDyn was unaffected at any time. Airway reactivity to MCh (as ΔRL) was increased at 4 h post‐exposure, and ΔCDyn responses were decreased. GS was essentially without effect on RL or CDyn at 1 d, 1 wk, 1 mo and 2 mo after administration. The results indicate that a single exposure to GS increases transiently lung resistance and reactivity to MCh.Grant Funding Source: Supported by NIOSH
Wheel access in the prior 22 hours is sufficient to reduce d‐methamphetamine self‐administration in rats (661.5)
Authors: Taffe Michael , Aarde Shawn , Creehan Kevin , Vandewater Sophia 
Published: June 2021

Initial rat studies appeared to show that an extended, ~6 week interval of activity wheel exposure was needed to reduce intravenous self‐administration of stimulant drugs. Yet a recent study found that ~22hrs of wheel access prior to sessions, without any extended prior exposure, was sufficient to reduce cocaine self‐administration in a between‐groups design. The present study sought to distinguish if the effects of ~22hr wheel access on methamphetamine (MA) self‐administration are an acute or lasting phenomenon using a within subjects design. Groups of male Sprague Dawley rats were trained to stably self‐administer MA (0.05 mg/gk/infusion, i.v.) and then subjected to wheel access/no access conditions when in the vivarium. Intake of MA was significantly lower on sessions following ~22 hrs of access to the wheel compared with sessions which followed ~22 hrs of no‐wheel housing. Effects of the wheel were observed within‐subject in an ABAB design as well as between groups. These data provide evidence that a suppressive effect of wheel activity that had previously been attributed to sustained, multi‐week wheel access is instead due to acute wheel activity in the day prior. Thus, brain plasticity mechanisms that require 6 weeks of chronic activity to produce are unlikely to be related to the suppressive effect of activity on drug self‐administration. This study also has important implications for using exercise as an adjunct to human drug cessation therapy because an extended interval of escalated activity may not be required. Likewise, consistent daily activity may be required for maximum effect.Grant Funding Source: Supported by USPHS Grant DA024105 and DA024705
KK‐92A, a novel GABAB receptor positive modulator, attenuates the rewarding effects of nicotine in rats (661.9)
Authors: Li Xia , Kaczanowska Katarzyna , Sturchler Emmanuel , Cameron Michael , McDonald Patricia , Finn M. G. , Griffin Patrick , Markou Athina 
Published: June 2021

GABAB receptors (GABABR) are thought to be one of the most promising targets for the treatment of drug dependence. GABABR positive allosteric modulators (PAMs) have been shown to exhibit similar effects as agonists in animal models of nicotine dependence with superior side‐effect profiles. This study involved the synthesis of new selective GABABR PAMs, and the assessment of their effects in animal models of nicotine dependence. New derivatives of BHF177, a potent GABABR PAM with efficacy in animal models of nicotine dependence, were synthesized. Nine active molecules were selected based on in vitro microsomal stability and efficacy in multiple cell‐based functional assays, including cAMP accumulation, Ca2+ mobilization, ERK activation and a cellular impedance‐based (CellKey) assay. Among these compounds, KK‐92A showed improved efficacy in the cAMP and Cellkey assays compared to BHF177, and had a similar in vivo drug metabolism and pharmacokinetic profile, thus, was selected for in vivo assessment. We found that KK‐92A decreased intravenous nicotine self‐administration under fixed‐ and progressive‐ratio schedules of reinforcement in rats. The results indicate that KK‐92A, a novel selective GABABR PAM, inhibited both the primary reinforcing and incentive motivational effects of nicotine, suggesting that GABABR PAMs may be useful antismoking medications.Grant Funding Source: NIH 2U19DA026838
G protein contribution to complex signaling pathways downstream of the dopamine D2L receptor (662.10)
Authors: Brust Tarsis , Hayes Michael , Roman David , Watts Val 
Published: June 2021

The dopamine D2 receptor (D2R) is a drug target that activates multiple signaling pathways via G protein‐dependent and independent mechanisms. Different ligands can activate these signaling pathways with distinct potencies and efficacies (i.e. biased ligands) which may provide a path for developing safer and more effective therapies. The present study examined several complex signaling pathways that may require multiple immediate effectors of the D2R. Specifically, we analyzed ERK phosphorylation, heterologous sensitization, AKT phosphorylation, and dynamic mass redistribution in response to a series of D2R ligands. The ligands tested included previously characterized D2R modulators that are used for the treatment of Parkinson’s disease and schizophrenia. The results were compared to previous studies examining Gαi/o, Gβγ, and β‐arrestin signaling in the same model system. Ligand bias was assessed using previously described methods as well as a new Weighted Efficacy Analysis (WEA). The WEA utilizes a straightforward equation incorporating two key pharmacological measures (i.e. pEC50 and relative efficacy) to calculate bias. Additional studies explored the requirement of both Gαi/o and Gβγ activation for these complex signaling pathways. Our results reveal unique bias profiles of several D2R ligands and provide insight regarding the effector requirements for complex signaling pathways of the D2R.Grant Funding Source: Supported by MH60397 and BBRF.
Expression of olfactory GPCR signaling genes in the eye (662.15)
Authors: Slepak Vladlen , Pronin Alexey , Levay Konstantin , Velmeshev Dmitry , Faghihi Mohammad , Shestopalov Valery 
Published: June 2021

Ocular surface is continuously exposed to particles, microorganisms and other stressors. To advance our understanding how the outer eye interacts with its environment, we asked which cellular receptors are expressed in the cornea, focusing on identification of G protein‐coupled receptors (GPCRs). Total RNA was isolated from the mouse cornea and subjected to next‐generation sequencing using Illumina platform. The data was analyzed by CuffLinks and TopHat software. Gene expression was confirmed by RT‐PCR. Cellular localization of transcripts was analyzed by in situ hybridization using RNAscope technology. The determined mouse cornea transcriptome contained more than 45 million reads representing over 10,000 gene transcripts. We identified more than 150 GPCR gene transcripts including more than 100 were putative olfactory receptors (Olfr’s). Our RT‐PCR analysis confirmed the presence of several Olfrs and related genes, for example, the G protein associated with olfaction, Galpha‐olf, and olfactory marker protein (OMP). In situ hybridization showed that mRNA for Olfr558 was present primarily in vessels of the eye, whereas OMP was found in epithelial cells of the cornea. Thus, our data show that several species of mRNA encoding putative olfactory receptors and related genes are expressed in the mouse cornea and other parts of the eye.Grant Funding Source: Supported by NIH RO1EY018666 (VZS), R01EY017991 (VIS), P30 EY014801
Death effects of reveromycin A in chondrocytes and synoviocytes (663.14)
Authors: Svrcina Haley , Steinke Ellen , Davitt Kelsey , Sloane Emily , Williams Calli , Jones Dan 
Published: June 2021

Osteoporosis is a prevalent disease with several current treatments. However, these treatments can have harmful long‐term side effects on individuals utilizing them. Reveromycin A (Rev A) is a natural compound that specifically causes apoptosis in osteoclasts but not osteoblasts. To further test whether Rev A can be a safe osteoporosis treatment without detrimental effects in the surrounding joint, the compound’s ability to induce apoptosis and necrosis in ATDC5 murine chondrocytes and HIG‐82 rabbit synoviocytes was measured. Inflammatory joint disease is known to accompany osteoporosis; consequently, Rev A‐induced cell death was characterized in each of these two cell lines under both physiological pH and a pH of 6.8 to mimic the pH of an inflamed joint. After treatment, morphology was assessed through phase contrast microscopy, apoptosis was measured through colorimetric caspase 3 activity assay, and necrosis was quantitated by use of a lactate dehydrogenase (LDH) release assay. Six hours of ten micromolar Rev A treatment did not significantly induce chondrocyte apoptosis, chondrocyte necrosis, or synoviocyte necrosis at either pH. Synoviocyte apoptosis was not induced by Rev A at physiological pH, but at a pH of 6.8, Rev A induced apoptosis compared to untreated by 4.9–fold (p<0.005). The results suggest that Rev A may have undesirable apoptotic effects in synoviocytes of the joint under acidic conditions.Grant Funding Source: Supported by grants from the Indiana Academy of Science and the Hodson Summer Research Institute
Reveromycin A‐induced apoptosis in osteoclasts is accompanied by decreased NF kappa B levels (663.15)
Authors: Jones Dan , Mead Brittany , Morgan Heather , Tedeschi Laura , Sloan Chris , Lang Spenser , Hines Cory , Gragg Megan , Stofer Jonathan , Riemann Kaitlin , Derr Tyler , Heller Emily , Collins David , Landis Paul , Linna Nathan 
Published: June 2021

Reveromycin A (Rev A), a small natural product isolated from Streptomyces bacteria, is a potential osteoporosis therapeutic in that it specifically induces apoptosis in osteoclasts but not osteoblasts. The purpose of the study presented here was to further elucidate the intracellular mechanisms of Rev A death effects in mature osteoclasts. RAW264.7 murine macrophages were differentiated in the presence of receptor activator of nuclear factor kappa B ligand (RANKL) and stained for tartrate‐resistant acid phosphatase to confirm their osteoclast character. These osteoclasts were treated with ten micromolar Rev A for 2, 4, and 6 hours at a pH of 5.5. Peak apoptosis induction occurred at 4 hours (5.1‐fold, p<0.05) as measured by caspase 3 activity. Lactate dehydrogenase release assay revealed no significant Rev A‐induced necrosis. Western blot analysis of cytoplasmic extracts demonstrated activation of caspase 9 and a mild decrease of deamidated Bcl‐XL. In both cytoplasmic and nuclear extracts, NF kappa B levels increased on differentiation with RANKL but then decreased at 2 hours of treatment (2.4‐fold for cytoplasmic and 1.7‐fold for nuclear levels). Rev A‐induced apoptosis in osteoclasts is not accompanied by necrosis, suggesting that Rev A would likely have limited effects on neighboring bone cell types, and Rev A decreases NF kappa B, a factor needed for differentiation and survival of the mature osteoclast.Grant Funding Source: Supported by an Indiana Academy of Science grant and a Hodson Summer Research Institute grant
MAGI1 signaling in cerebral cavernous malformations (664.11)
Authors: Sanchez Atlee , Stockton Rebecca 
Published: June 2021

Cerebral Cavernous Malformations (CCMs) are vascular brain lesions affecting 0.5% of humans. Late stage lesions are clusters of grossly dilated capillaries lacking smooth muscle cell or pericyte support, and can cause hemorrhagic stroke. No treatment exists other than surgical removal of symptomatic lesions. CCMs occur sporadically through spontaneous mutations, or by familial autosomal dominant transmission of mutations in three genes : KRIT1, CCM2 or CCM3 (PDCD10). Our lab’s previous work showed that KRIT1 and CCM2 normally regulate endothelial permeability through regulation of RhoA and ROCK1/2. We now used proteomics and CHO cell co‐immunoprecipitations to identify a signalosome consisting of a unique brain microvascular endothelial cell‐specific splice variant of Rap1‐interacting junctional protein MAGI, “MAGI1C‐beta2”, that binds to KRIT1 and CCM2 through a PDZ domain to mediate translocation of two CCM proteins to endothelial junctions, and is required to maintain barrier function. We further identified a MAGI‐1 interacting protein , AMOT (angiomotin) as a convergence point for all three CCM protein signaling pathways, by showing that PDCD10 interacts directly with AMOT at junctional complex , while KRIT1 and CCM2 are localized there through MAGI1C‐beta2. Further, this intact CCM1‐2‐3/AMOT complex suppresses RhoA activation, providing a mechanistic explanation for some aspects of CCM lesion development.Grant Funding Source: NIH