bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Key Transcription Factor Target Interactions That Regulate Prostate Cancer Metastasis
Authors: Sharma Nitya , Pellegrini Kathryn L. , Giuste Felipe O. , Ouellet Veronique , Trudel Dominique , Mes‐Masson Anne‐Marie , Saad Fred , Osunkoya Adeboye O. , Petros John , Moreno Carlos S. 
Published: November 2023

Prostate cancer remains the most commonly diagnosed cancer in U.S. males, and ranks second in mortality with over 28,000 deaths per year. The standard of care for patients with recurrent, aggressive prostate cancer is androgen deprivation therapy (ADT), but the benefits from ADT are typically short‐lived. Recurrent disease following ADT treatment is termed castration‐resistant prostate cancer (CRPC), and is generally incurable after progression to metastatic disease. Therefore, understanding the mechanisms underlying CRPC and subsequent progression to metastatic disease is critical. To gain insights into how transcriptional networks change in response to ADT and lead to metastasis, we have identified the relationships between transcription factors and corresponding gene targets in matched pre‐ADT and post‐ADT tissue samples, as well as matched primary and metastatic lesions. First, we generated gene expression data by sequencing RNA from 24 formalin‐fixed paraffin‐embedded patient‐matched pre‐ADT needle core biopsies and corresponding post‐ADT radical prostatectomy prostate cancer samples. For generating metastatic networks, we used publicly available expression data from matched primary prostate and metastatic tumors. Next, we integrated mRNA expression, protein‐protein interaction, and DNA binding motif data using the PANDA algorithm to reverse engineer transcriptional networks. We identified key transcription factors with significant gains or losses of interactions with target genes specifically in metastatic networks. We also identified putative novel key prostate cancer specific transcription factor interactions that share multiple gene targets, such as ETS1 and GATA2, possibly revealing relationships that directly contribute to increased metastatic potential. By comparing changes in post‐ADT and metastatic transcriptional networks, we may identify critical transcription factor‐target gene interactions that are essential for the progression of CRPC to metastatic disease. This study may thus provide insights into novel therapeutic approaches for treatment of CRPC and prevention of metastasis, as well as prognosis of patients with poor outcomes.Support or Funding InformationMovember Foundation
Beneficial effect of exogenously applied melatonin on wound healing in skin injured animal model
Authors: Choi Jeonghyun , Won Jinyoung , Hong Yonggeun 
Published: November 2023

Skin injury (SI) is generated by various situations such as surgery, burning etc.. Many factors are participated in wound healing processes. The present study was conducted to investigate the effect of exogenously applied melatonin (MT) on the wound healing. In this study, subjected Sprague‐Dawley rats were subdivided into three groups; Con (control), Veh (SI+Saline), MT (SI+MT). Several genes related to inflammation were analyzed in mRNA using RT‐PCR. The expression of TNF‐α was significantly increased in Veh compared to MT (p<.05), whereas, TNF‐R2 expression was higher than in MT (p<.05). Also, in MT group, the expression of p53 and CHOP as apoptosis marker was significantly increased compared to Veh (p<.05). IGF‐1, IGF‐1R, TGF‐β1, and COL1 expression related to collagen synthesis showed similar pattern. Several markers that related to angiogenesis were analysed from two wound margins. HIF‐1α and VEGF expression was significantly increased in MT compared to Veh (p<.05). Also, exogenous melatonin impacted on the upregulated expression of GPx in MT. We observed fibroblast like fibers, adipocytes, blood vessels, collagen like fibers, and hair follicles in H&E staining. Also, wound closing rate was significantly increased in MT compared to Veh at 2, 7, 10, and 21 day (p<.05). Taken together, it is suggested that exogenously applied melatonin might contribute to the skin restoration via regulation of various factors related to inflammation, collagen synthesis, and angiogenesis.Support or Funding InformationFunds: NRF‐2013R1A2A2A01067169 to Y.H., 2014R1A1A3051724 to S.P., KRIBB Research Initiative Program (KGM4611613 to Y.H.), Republic of Korea
Inactivation of the glyoxalase anti‐glycation system in porcine brain post‐cardiac arrest and its preservation by pyruvate
Authors: Scott Gary Frank , Cherry Brandon H , Williams Arthur , Ryou Myoung‐Gwi , Nguyen Anh , Mallet Robert T 
Published: November 2023

BackgroundGlyoxalase‐1 (GLO1) degrades the toxic glycolytic byproduct methylglyoxal, thereby preventing irreversible glycation of proteins and nucleic acids. However, the glyoxalase system may be inactivated by oxidative stress. Cardiac arrest (CA) and cardiocerebral resuscitation (CCR) provokes intense oxidative stress in the brain. However, the impact of CA/CCR on the brain's anti‐glycation machinery is unknown.PurposeTo analyze the impact of ischemia‐reperfusion on the cerebrocortical glyoxalase system in a porcine model of CA/CCR, and to determine the ability of the natural antioxidant pyruvate to protect this crucial anti‐glycation system.MethodsDomestic pigs (25–35 kg) were subjected to 10 min CA, followed by 4 min closed‐chest CCR, defibrillation and 4 h recovery. Sodium pyruvate (Figures: Pyr) or NaCl control was infused iv to 3.5–4 mM during CA/CCR and the first 60 min recovery. GLO1 content, activities of GLO1 and glutathione reductase (GR), and protein glycation (Arg‐pyrimidine immunoblot) were analyzed in frontal cortical homogenates collected at 4 h recovery.ResultsCompared with non‐arrested sham pigs (n=4), GLO1 content and GLO1 and GR activities (Figures 1–3) fell sharply after CA/CCR (n=4), but were preserved by pyruvate (n=4). Also, CA/CCR caused protein glycation, particularly at 65 kDa, the MW of the blood‐brain‐barrier component occludin, while pyruvate reversed this glycation.ConclusionsCardiac arrest‐resuscitation impaired the brain's GLO1/GR anti‐glycation defenses in a manner prevented by pyruvate treatment. Thus, the brain's GLO1 system is disabled by acute ischemia‐reperfusion, yet pyruvate intervention protected this pivotal, cerebroprotective system.Support or Funding InformationNS076975 National Institute of Neurological Disorders and Stroke
Comparison of Topical PDGF and Naltrexone on Full‐Thickness Diabetic Wound Healing
Authors: Titunick Michelle Beth , Cain Jarrett , Zagon Ian , McLaughlin Patricia 
Published: November 2023

Diabetes affects more than 29 million individuals in the US, and approximately 15% will develop chronic, non‐healing foot ulcers (DFU) that may result in amputation if left untreated. The current FDA‐approved DFU treatment is Regranex® which increases platelet derived growth factor (PDGF), but is expensive and has significant side‐effects. A new topical treatment utilizing naltrexone (NTX) to block classical opioid receptors as well as Opioid Growth Factor Receptor (OGFr) has been shown to accelerate full‐thickness cutaneous wounds in preclinical models of type1 and type 2 diabetes by increasing cell replication and angiogenesis. NTX blocks interaction of the inhibitory peptide OGF and OGFr; OGF is elevated in serum of diabetic humans and animals. To compare the efficacy of NTX and Regranex®, type 1 diabetes was induced in male Sprague Dawley rats (n=14) by injection of streptozotocin (40mg/kg i.p. on two consecutive days); control rats received vehicle only (n=13). Five weeks later, four 6 mm wounds were produced on the dorsum of each animal; wounds were randomly assigned treatment of PDGF (Regranex®), 0.03% NTX dissolved in moisturizing cream, or cream containing sterile saline (control). Residual wound size was photographically monitored over a 2 week period. DNA synthesis (BrdU integration), as well as PDGF and vascular epithelial growth factor (VEGF) expression were measured in wound tissue (3–4 rats per group) collected 1,2 and 4 days following surgery. Both NTX and Regranex® significantly accelerated wound closure in comparison to the vehicle‐treated wounds. By day 4 following surgery, there was a 1.2‐fold decrease in residual wound size in rats treated with either NTX or Regranex® relative to vehicle‐treated wounds, and by day 12, NTX‐treated wounds were reduced in size by 5‐fold relative to vehicle‐treated wounds. All wounds were <1% the original size by day 14.Four days post‐wounding, DNA synthesis was elevated 43% in diabetic rats receiving NTX relative to diabetic vehicle‐treated rats (p≤0.01). Immunohistochemical examination of wounds one day after surgery revealed that Regranex®‐treated diabetic rats had 139% more PDGF positive cells than vehicle‐treated diabetic rats (p≤0.01), whereas NTX‐treated diabetic rats exhibited approximately 78% increase in PDGF+ cells compared to vehicle‐treated rats (p≤0.05). At 2 days, NTX‐treatment resulted in a 55% increase in PDGF+ cells relative to controls, whereas the Regranex and controls were comparable. Four days after surgery there was a 44% increase in VEGF+ vessels in NTX‐treated diabetic rats (p≤0.001) relative to those receiving vehicle only and a 69% increase when compared to diabetic Regranex®‐treated rats (p≤0.001). FGF immunohistochemistry did not indicated significant differences in expression at wound sites between any treatment groups. The data suggest that blockade of the OGF‐OGFr regulatory pathway utilizing NTX may be a safe and effective alternative treatment to Regranex® for DFUs.
Protective Role of Metallothioneins in Human Endothethial Cells in Acute Response to High Dose Cadmium Chloride
Authors: Zhang Yanmin , Kucynda Theresa T. , Marsboom Glenn , Kajdacsy‐Balla Andre 
Published: November 2023

IntroductionCadmium is a well‐recognized environmental pollutant with numerous adverse health effects. Recent research has demonstrated the association of cadmium with cardiovascular disease through endothelial dysfunction at substantially low levels of chronic exposure. However, little is know about the effect of high concentration cadmium effect on human endothelial cells after acute exposure.MethodsPrimary human umbilical vein endothelial cells (HUVECs) were treated with varying doses of cadmium chloride (CdCl2). Flow cytometry was used to evaluate the cells reactive oxygen species (ROS) with cell‐permeant 2′,7′‐dichlorodihydrofluorescein diacetate (H2DCFDA) probe. The oxygen consumption rate (OCR) of adherent cells was measured at 37°C using a Seahorse XF24 Extracellular Flux Analyzer. Gene expression was measured with real‐time RT‐PCR.ResultsThe HUVECs cells were treated with varying dose of CdCl2 for 24 hours and cells viability was not changed at any of the dose used. Real time RT‐PCR data revealed both metallothionein‐1A (MT1A) and metallothionein‐2A (MT2A) gene levels were significant increased by 50 μM CdCl2 compared to the control levels (13.8‐fold and 5.4‐fold increase, respectively, P<0.05). OCR of HUVECs was decreased by 50 μM CdCl2 treatments from 66.97±2.41 to 42.00±5.22 pMoles/min (P<0.005), and H2DCFDA fluorescence intensity was decreased from 2989±87.75 in control to 2165±54.75 (P<0.005). However, with lower concentrations, short‐term exposure did not cause significant changes in ROS level and oxygen consumption rate. We are currently exposing cells to lower concentrations for longer period of time.ConclusionsIt is generally agreed that oxidative stress plays important roles in acute cadmium poisoning. Interestingly, our data demonstrated decreased oxygen consumption and lower ROS level upon acute high dose cadmium exposure. The possible mechanism could be that during acute response to high dose cadmium, metallothioneins (MTs) are induced and act as a potent sequestration mechanism, thus protecting against oxidative stress.
Mode of the inhibition of venom collagenase by actinonin
Authors: Price Joseph A. , Sun Jing 
Published: November 2023

Snake envenomation remains an endemic health problem in much of the world. A new approach to controlling its pathology may be the use of enzyme inhibitors. A preliminary characterization was made of the mode of C. atrox collagenase inhibition by the protease inhibitor actinonin. The reaction progresses nearly linearly for close to an hour at room temperature. Reaction data for combinations of seven levels of inhibitor and six levels of substrate were fit to Michaelis‐Menten equations by nonlinear regression to determine values of apparent Km and Vmax. In these dose response experiments apparent Vmax, Km, and Vmax/Km decreased as the concentration of the inhibitor was increased. This pattern is most consistent with noncompetitive enzyme inhibition. These results better characterize this agonist in reactions with whole venom, and make actinonin a compound of interest in developing new anti‐venom therapeutics.Support or Funding InformationNo extramural funding supported this work.
Location, Location, Location: The Effect of Lorcaserin via Different Routes of Administration on Acute Oxycodone Analgesia
Authors: Lippold Kumiko , Dewey William L. 
Published: November 2023

Pain is an unpleasant, subjective experience that serves as an important defense mechanism to noxious stimuli. There are several classes of analgesic drugs prescribed for the treatment of pain, with the most well‐known being opioids. Opioids are almost exclusively indicated for the treatment of moderate to severe pain but with their repeated use, there is a development of tolerance and dependence. With that known, there has been great interest to develop new analgesics or drug combinations that may reduce the incidence of side‐effects and facilitate the analgesic effects. Lorcaserin is a 5‐HT2C agonist that was originally indicated as a treatment for obesity but hypothesized as a potential treatment for drug dependence. Several preclinical studies have shown that lorcaserin reduced the self‐administration of several classes of drugs of abuse, including opiates. This observation prompted us to investigate whether lorcaserin would alter the basic pharmacology of opioids. To test this, we used a preclinical model (tail immersion test) that utilizes an acute, noxious thermal stimulus and is a heavily validated method for testing the antinociceptive properties of opiates. In the first round of studies we investigated the effect of subcutaneous lorcaserin pretreatment on the acute antinociceptive effect of oral oxycodone. All doses of lorcaserin tested produced a reversed dose‐response shift in the oxycodone curve (ED50 = 6.11mg/kg) to the left, with significant ED50 shifts occurring at 0.25mg/kg (ED50 =1.3mg/kg), 0.5mg/kg (ED50 = 1.21mg/kg), 1mg/kg (ED50= 2.08mg/kg), and 2mg/kg of lorcaserin (ED50=3.77mg/kg). It is important to note that none of the subcutaneous doses of lorcaserin tested had an antinociceptive effect on their own. Next, we administered lorcaserin via different routes of administration and found that its effects vary depending upon route of administration. Lorcaserin, administered via the intracerebroventricular route at 32, 64 or 128ug did not demonstrate antinociceptive activity but intrathecal lorcaserin (8, 16, 32, 64, 128ug) demonstrated a full dose‐responsive effect, with an ED50 of 54ug. Interestingly, we observed that ICV lorcaserin attenuated oral oxycodone's antinociceptive effects by 40%. Our results indicate that there are significant regional differences in the effect of lorcaserin and the 5‐HT2c receptor plays a differential role depending on the region of the central nervous system in the antinociception elicited by oxycodone. Further studies will investigate the role of potential off‐target effects of lorcaserin that may be mediated through the 5‐HT2A receptor.Support or Funding InformationFunding provided by USPHS grant number DA036975.
Regulation of Cytoskeletal Remodeling Proteins in the Ventral Tegmental Area by Morphine, Stress, and TORC2
Authors: Kaska Sophia , Brunk Rebecca , Kechner Megan , Mazei‐Robison Michelle 
Published: November 2023

Drug addiction and depression are two co‐morbid diseases that have a substantial burden on society. Increasing evidence suggests that dysfunction of the reward circuit could underlie both disorders. In support of this, chronic morphine and chronic social defeat stress (CSDS) have been shown to induce similar changes in ventral tegmental area (VTA) DA neuronal morphology. Specifically, our previous work demonstrated a morphine‐induced decrease in VTA DA size that was mammalian target of rapamycin complex 2 (TORC2) signaling‐dependent. Similarly, preliminary evidence from our lab suggests that mice that are susceptible to CSDS also have a decrease in VTA DA soma size. Our current objective is to identify the molecular mechanisms underlying these morphological changes. Given our data that morphine‐induced changes in VTA DA soma size are TORC2‐dependent and recent evidence that TORC2 signaling modulates cytoskeletal remodeling in the hippocampus, we hypothesize morphine and stress may alter the actin cytoskeleton in VTA DA neurons through alteration of TORC2 activity. We completed western blot analyses on micro‐dissected VTA tissue from mice exposed to chronic morphine or CSDS and found a significant decrease in the phosphorylation of cofilin, a protein involved in severing actin filaments. We are now examining molecules upstream of cofilin, such as Rac1 and p‐21 activated kinase (PAK) to determine if exposure to chronic morphine or stress similarly alters activity of these proteins. Additionally, to further investigate the potential role of TORC2 on mediating changes in the actin cytoskeleton, we are performing western blot analysis using VTA tissue from mice that either overexpress or knockout Rictor, a critical component of TORC2. Overall, our goal is to determine the molecular mechanisms underlying changes in VTA structural plasticity in the hopes of identifying novel targets for therapeutic intervention in addiction and depression.Support or Funding InformationPhRMA Foundation Pre‐Doctoral Fellowship in Pharmacology/Toxicology (SK)Integrative Pharmacological Sciences Training Program: T32 GM092715 (SK)NIDA: R01 DA039895‐01A1 (MMR)NIDA: R03 DA037426 (MMR)PhRMA Foundation Research Starter Grant (MMR)
The Role of Norbuprenorphine in Buprenorphine‐Induced Neonatal Abstinence Syndrome: A Pilot Study
Authors: Brents Lisa K , Caperton Caitlin O , Patton Amy L , Owens S Michael , Moran Jeffery H , Stowe Zachary N , Fantegrossi William E 
Published: November 2023

Opioid abuse during pregnancy may cause drug dependence in the fetus, and the abrupt discontinuation of opioid exposure at birth may induce neonatal abstinence syndrome (NAS) during the first week of life. NAS is characterized by inconsolable high‐pitched crying, tremor, vomiting, diarrhea, hypersensitivity, and sleep disturbances. Buprenorphine (BUP), a mu‐opioid receptor partial agonist, is administered as a substitution therapy to treat opioid use disorder during pregnancy. BUP improves neonatal outcome compared with methadone or no treatment, but BUP exposure often leads to NAS. Furthermore, NAS severity is independent of maternal BUP dose throughout pregnancy, suggesting that individual differences in maternal‐fetal BUP pharmacology may increase NAS susceptibility in some newborns. One possible explanation is that fetal exposure to norbuprenorphine (NorBUP), a metabolite of BUP that has high affinity and efficacy at the mu‐opioid receptor, differs across the BUP‐treated population and drives individual differences in the development of opioid dependence. This study tested the hypothesis that NorBUP can induce NAS. To test this hypothesis, we developed a novel model of NAS in which exposure to opioids occurs continuously across fetal development. Pregnant rats were implanted with subcutaneous minipumps containing NorBUP (0.1 or 0.3 mg/kg*day) or vehicle (5 % Emulphor) on gestation day (GD) 9. Treatment continued until delivery. Upon delivery, half of the pups were observed and scored for signs of precipitated withdrawal for 10 minutes after administration of the mu‐opioid receptor antagonist naltrexone (1 or 10 mg/kg, i.p.) or saline (10 ml/kg). Because of the CNS immaturity of the neonatal rat pup relative to the human newborn, this group models opioid dependence in preterm newborns. To model NAS at term, the other half of each litter was fostered to an untreated dam and received subcutaneous injections of NorBUP (0.05 mg/kg or 1.5 mg/kg) every 12 hours (BID) until they were tested for precipitated opioid withdrawal on postnatal day (PD) 7. Few withdrawal signs were observed in the pups tested within 24 hours of delivery, but PD7 pups exposed to the high dose of NorBUP in utero and after birth exhibited significantly more “wall climbing” and less “quiet” signs when challenged with naltrexone, indicating opioid withdrawal. The results of these preliminary studies suggest that NorBUP can induce opioid dependence in the neonatal pup. Ongoing experiments will determine fetal brain concentrations of NorBUP after maternal exposure to NorBUP (GD 9‐ GD20). Results of this experiment will be used to determine additional doses of NorBUP to be tested in the novel rat model of NAS.Support or Funding InformationSupport for this study was provided by the UAMS Fellow to Faculty award and the UAMS Tobacco Research Committee Tobacco Settlement Funds
Depression of home cage wheel running reveals greater spontaneous morphine withdrawal in rats with persistent pain compared to naïve controls
Authors: Lee Andrea T , Kandasamy Ram , Morgan Michael M 
Published: November 2023

The widespread use of opioids to treat pain has led to a drastic increase in opioid dependence and overdose fatalities. The transition from the use of opioids to treat pain to abuse is driven in part by withdrawal symptoms associated with the abrupt use of opioids. The objective of the present study is to develop an animal model of withdrawal that mimics the spontaneous withdrawal that occurs in pain patients. We hypothesized that spontaneous opioid withdrawal will cause depression of home cage wheel running. This hypothesis was tested by measuring home cage wheel running before and after subcutaneous implantation of two 75 mg morphine or placebo pellets into male Sprague‐Dawley rats with and without hindpaw inflammation. Administration of the morphine pellets depressed wheel running that recovered to baseline levels by the third day after implantation in naïve rats and by the fifth day after implantation in inflamed rats. Removal of the morphine pellets depressed wheel running for 2 days in naïve animals and over 6 days in rats with an inflamed paw. These data demonstrate that home cage wheel running is a consistent and reliable method to assess spontaneous morphine withdrawal in the rat. Moreover, the magnitude of withdrawal is greatly enhanced in rats with persistent pain.Support or Funding InformationThis study was supported in part by State of Washington Initiative Measure No. 171 (to ATL) and NIH grant NS095097 (to MMM).