bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Effect of Cardiolipin on Microglial Activation and Microglia‐Mediated Neuronal Death
Authors: Pointer Caitlin , Klegeris Andis 
Published: November 2023

Microglia, the immune cells of the brain, become activated in response to a variety of stimuli. Under normal physiological conditions, microglial activation is advantageous, as it produces an acute inflammatory response that helps rid the brain of the noxious stimuli through the release of pro‐inflammatory mediators and cytotoxins, including reactive oxygen species (ROS) and reactive nitrogen species (RNS). When microglial activation is sustained, however, the persistent and non‐specific release of these cytotoxic molecules results in damage to surrounding cells, including neurons and other microglia. Such state of chronic neuroinflammation is a characteristic feature of Alzheimer's disease (AD). Cardiolipin (CL), a phospholipid located in the inner mitochondrial membrane, is significantly reduced in AD brains. The remaining CL is exposed to the high levels of ROS present in AD brains, which modifies the structure of CL and hinders its ability to regulate mitochondrial processes. This results in the further production of ROS and extensive neuron death. The intracellular role of CL is well defined; however, the effects of extracellular CL on microglial activation, and subsequently on neuronal viability, have not been described.We utilized cell culture techniques to study the effects of extracellular CL on the activation of microglia, and their subsequent toxicity towards neurons. BV‐2 murine microglia, as well as human HL‐60 promyelocytic and THP‐1 monocytic cells (microglia models), were exposed to CL in the presence of noxious stimuli for 24 or 48 hours. Pro‐inflammatory mediators including tumor necrosis factor (TNF)‐alpha, monocyte chemoattractant protein (MCP)‐1, ROS and RNS in supernatant were quantified to determine the effect CL has on the secretory profile of activated microglia. Additionally, THP‐1 cell supernatants were transferred onto human SH‐SY5Y neuroblastoma cells (neuron model) to determine whether CL regulates microglia‐mediated neurotoxicity.We discovered that CL decreased the secretion of TNF‐alpha, MCP‐1, ROS and RNS from activated microglial cells. Additionally, CL reduced the cytotoxicity of activated THP‐1 cells towards SH‐SY5Y neuronal cells.These findings indicate that CL may act as a novel intercellular signaling molecule that is capable of reducing microglial activation, as well as neuron death. Further studies will explore glial signaling pathways engaged by CL and reveal its role in neuron‐microglia communication.
High Fat Diet Increases Cognitive Decline and Neuroinflammation in a Model of Orexin Loss
Authors: Duffy Cayla Marie , Nixon Joshua P , Butterick Tammy A 
Published: November 2023

Midlife obesity is a risk factor for cognitive decline and is associated with the earlier onset of Alzheimer's disease (AD). Diets high in saturated fat can potentiate the onset of obesity, microglial activation, and neuroinflammation. Signaling deficiencies in the hypothalamic peptide orexin A (OXA) and/or orexin fiber loss are linked to neurodegeneration in humans and animal models. Prior studies in orexin/ataxin‐3 (O/A3) mice, a transgenic mouse model of orexin neurodegeneration, demonstrate that OXA treatment improves cognitive processes. In separate studies we have demonstrate that OXA can suppress neuroinflammation and is neuroprotective against the saturated fatty acid palmitate. Our overall hypothesis is that OXA treatment attenuates hippocampal neuroinflammation and prevents cognitive decline in obesity during high fat diet (HFD) feeding. Our short term goal for this project is to determine if OXA loss increases neuroinflammation and impairs cognition during HFD feeding. To examine this, we tested male O/A3 mice (7–8 mo. of age) in a two‐way active avoidance (TWAA) hippocampus‐dependent memory task. We tested whether 1) OXA loss impaired cognitive function; 2) HFD worsened cognitive impairment; and 3) HFD increased microglial activation and neuroinflammation. We show that O/A3 mice have significant impairments in TWAA task learning vs. wild type (WT) mice (increased escapes p<0.05, reduced avoidances p<0.0001). Mice were then placed on HFD (45% total fat, 31.4% saturated fat) or remained on normal chow (NC; 4% total fat and 1% saturated fat), and TWAA was retested at 2 and 4 weeks. Learning impairment was evident at both 2 and 4 weeks in O/A3 mice fed HFD for following diet exposure vs. WT mice on normal chow or HFD (increased escapes, reduced avoidances p<0.05). Additionally, O/A3 mice had increased gene expression of the microglial activation marker, Iba‐1 (measured via qRT‐PCR, p<0.001). Characterization of the microglial immune response (M1 neurotoxic vs. M2 protective phenotypes) is ongoing and will include gene and protein markers (such as TNF‐α, iNOS, UCP2, and Arg1). Collectively, our results indicate that OXA loss impairs memory, and that HFD accelerates hippocampus‐dependent learning deficits and the onset of neuroinflammation. Future studies will focus on restoration of OXA within hippocampal circuits to reduce neuroinflammation and improve cognitive functionSupport or Funding InformationThis work was funded by the US Department of Veterans Affairs BLR&D IK2 BX001686 and IS1 BX003083A (to TAB), the University of Minnesota Healthy Foods, Healthy Lives Institute (to JPN and TAB), MnDRIVE Brain Conditions Fellowship in Neuromodulation (to CMD), and the Minnesota Veterans Medical Research & Education Foundation (to TAB).
Inhibition of Stroke‐Induced Injury by Phlebotomy
Authors: Tsai Yung‐Hsu , Liou Houng‐Chi , Tang Sung‐Chun , Liou Horng‐Huei , Jeng Jiann‐Shing , Fu Wen‐Mei 
Published: November 2023

Stroke is the second most common death and a leading cause of long‐term disability worldwide, yet the therapeutic options are still limited. Hemodilution has been reported to reduce brain infarction in experimental stroke model. Phlebotomy causes hypovolemic‐hemodilting effect, but whether it offers protection after ischemic stroke remains unclear. Using permanent occlusion model, we aimed to assess the effect of phlebotomy on ischemic stroke.Male spontaneously hypertensive rats (SHR) were subjected to permanent MCA occlusion (pMCAO) to produce cerebral ischemia. Phlebotomy was conducted by drawing 0.8 ml of whole blood after the pMCAO. In another set of experiment, macrophage migration inhibitory factor (MIF, 10 or 20μg/kg, i.v.) or its antagonist, ISO‐1 (3 mg/kg, s.c.), were administered 3 h after pMCAO. Brain infarction, neurological function and protein expression were analyzed 24 h after pMCAO. Cerebrovascular integrity was analyzed 7 h after pMCAO. Analysis of serum by proteome profiler array and ELISA were performed at 3 h, 6 h and 24 h after pMCAO.Our results show that phlebotomy at 3 h after MCA occlusion significantly reduced stroke‐induced infarction and brain edema and shows an improvement in behavioral deficit. Inflammatory responses were decreased in phlebotomy‐3h group as the expression of inflammation‐related mediators were reduced. Analysis of the cytokine profile in serum led us to presume Macrophage migration inhibitory factor (MIF) as a possible mediator of the protection by phlebotomy. MIF aggravated stroke injury with a larger infarction and a more severe cerebrovascular disruption was observed, whereas, its antagonist ISO‐1 inhibited stroke‐induced infarction. In conclusion, phlebotomy offers protection when conducted 3 h after stroke onset and may exert its protective effect through the suppression of inflammatory cascade by interfering the balance of inflammatory mediators in serum.Support or Funding InformationThis work was supported by the Ministry of Science and Technology grants, 104‐2321‐B‐002‐033, Taiwan.
1,2,3,4,6 Penta‐O‐Galloyl‐β‐D‐Glucose attenuates LPS/IFNγ‐induced MCP‐5 and Pro‐MMP‐9 Cytokines Release in BV‐2 Microglia Cells by Inhibiting NFkB and MAPK Signaling Proteins
Authors: Mendonca Patricia F , Taka Equar F , Bauer David F , Soliman Karam F 
Published: November 2023

Neurodegenerative diseases are characterized by an inflammatory process and activation of microglia cells that produce pro‐inflammatory cytokines, which are hardly evident in the normal brain. Many natural compounds derived from plants have shown anti‐inflammatory activity, such as 1,2,3,4,6 Penta‐O‐Galloyl‐β‐D‐Glucose (PGG), which is a naturally occurring polyphenolic compound highly enriched in some medicinal herbs such as Rhus chinensis Mill and Paeonia suffruticosa. PGG has been shown to exhibit an antiinflammatory effect through the inhibition of many cytokines in different experimental models, but not in microglia cells. In the present study, we investigated the role of PGG as an antiinflammatory agent in LPS/IFNγ‐activated BV‐2 microglia cells. Mouse cytokine antibody arrays were used to assess the effect of PGG in the release of proinflammatory cytokines, and ELISA experiments were performed to validate the results from the arrays. PGG signaling pathway was evaluated with NFκB and MAP Kinase PCR assays for the target genes, and the effect of PGG on protein expression was evaluated by Western blot. The results obtained showed that PGG decreased the expression of monocyte chemotactic protein‐5 (MCP‐5) and pro‐matrix metalloproteinase 9 (Pro‐MMP‐9). Both cytokines are upregulated during the inflammatory process and have been shown to be involved in brain injury, inflammation, and neurodegeneration. PGG also exhibited an inhibitory effect on several signaling proteins involved in the NFκB (IRAK1, NFKB1, NOD1) and MAP Kinase signaling pathways (CDK2), which were validated by mRNA and protein expression. In summary, these findings show that PGG modulatory effects on the release of MCP‐5 and Pro‐MMP‐9 cytokines may be through the downregulation of proteins involved in the NFκB and MAP Kinase pathways.Support or Funding InformationSupported by NIH Grants G12 MD007582 and P20 MD 006738
Naturally, Occurring Neurotrophic Factor from Mu Bie Zi (Momordica cochinchinensis) seeds
Authors: Mazzio Elizabeth A , Georges Beatrice , Soliman Karam F 
Published: November 2023

The discovery of nerve growth factor (NGF) mimetics could have the potential application for Alzheimer's disease prevention and therapy. Previously, we have conducted a high‐throughput (HTP) screening of over 1100 nutraceutical products (herbs, vitamins, plant‐based phytochemicals, etc.) to determine if any dietary based substance could induce neurite outgrowth similar to 7S NGF at <200ug/mL in PC‐12 cells. Only a single biologically active with compelling NGF mimetic capacity in the seed of Mu Bie Zi, Momordica cochinchinensis (bitter cucumber) (MCS). The objective of the current study is to identify the bioactive substances within the ground seed pit by testing for NGF biological activity in the crude seed solvent extractions and purified proteins. The active protein fraction was subjected to one‐dimensional gel electrophoresis (native), gel staining, sectioning / excision, electro‐elution back into solution and re‐evaluation for NGF mimetic effects. Results obtained show that the NGF mimetic resided in a low abundant visually absent gel section at around 17 kD. The 17kD gel slice was excised, digested and run on a UPLC‐MS/MS –with a Q Exactive Hybrid Quadrupole ‐ Orbitrap Mass Spectrometer. The data were evaluated using X! Tandem, OMS, X Hunter and K‐score algorithms. Proteomic evaluation of the 17kD band was conducted showing the presence of 11S globulin subunit beta, napin, oleosin 18.2 kDa, trypsin inhibitor 1 MCoTI‐I /II (known to contain growth factor cystine knots) and Two Inhibitor Peptide Topologies TIPTOPs (isoforms 1,2,3,5,6), all with taxonomical identities inherent to the cucumber family (Momordica cochinchinensis, macrophylla, charantia, Cucumis sativus).Support or Funding InformationThis research was supported by the National Institute for Minority Health and Health Disparities of the National Institutes of Health through Grant Number G12 MD007582 and Grant Number P20 MD006738
A Team‐based Assignment to Integrate Basic Science and Pharmacotherapeutic Principles for Anticancer Agents
Authors: Jungsuwadee Paiboon , Sakharkar Prashant , Kurup Sonali 
Published: November 2023

ObjectiveTo implement and evaluate an active‐learning, team‐based assignment centered on anticancer agents for the integration of basic science and pharmacotherapeutic principles.DesignStudents enrolled in the integrated hematologic and oncologic disorders course were divided into 5–6 students/group. Each group was assigned a specific anticancer agent and was expected to answer a series of questions designed to measure students learning abilities and competencies, followed by a presentation to the class.AssessmentEach assignment was assessed using a grading rubric that was mapped to the 2013 Center for the Advancement of Pharmacy Education (CAPE) outcomes to assess whether expected abilities and competencies are being met within a PharmD curriculum. Student learning was evaluated using performance on the assignment and examination questions. Student perceptions of the assignment were assessed using a short survey.ResultAssessments for student learning abilities and competencies were performed based on the team‐based assignment and presentation as well as three examinations, in which the first exam was taken place prior to the team‐based assignment and in‐class activities. Overall mean score on examination 1 was 75%, examination 2 and examination 3 were 81% and 77%, respectively. When only questions related to drugs covered in the assignment were analyzed, we found that students performed better on examination 2 and 3 that were completed after the team‐based assignment vs. examination 1. Student performance on 14 of the 53 questions on examination 2 that were related to the drugs covered as part of the assignment was 93.7%. Students performed better on assignment related questions on examination 2 compared to unrelated questions. Student performance on 21 of the 61 questions on examination 3 that were related to the drugs covered as part of the assignment was 85.9%. Although, student performance was better on assignment related questions on examinations 2 and 3 compared to unrelated questions, this differences were not significant (p=0.253). Finally, Student perceptions of the effectiveness of the team‐based assignment were also positive and majority of the students (83%) agree or strongly agreed that the assignment improved their knowledge and understanding of anticancer agents and should.ConclusionThe assignment provided a focused review of basic science and pharmacotherapeutic principles and enabled integration of concepts relating to the therapeutic application of anticancer agents and management of anticancer agent mediated adverse effects. The assignment could contribute toward preparing students for the evolving role of the pharmacist in the management of cancer.
Tools enhance learning of Medical Pharmacology
Authors: Zhong Qing 
Published: November 2023

ObjectiveMedical pharmacology learning is challenging in a Caribbean Medical School. The medical pharmacology curriculum only has 12 weeks. We created extra‐lecture activities (tools) to help pharmacology learning.MethodsWe introduced practice of baroreflex and volume‐pressure loop by applying drugs to enhance cardiovascular pharmacology learning, and tables to help chemotherapy learning. We had pre‐lecture and post lecture quiz, clinical case study, and team‐based learning (TBL) throughout the whole pharmacology course. We did a questionnaire to investigate how helpful of these tools in 2015 and 2016 after students finished the course. Students were asked to rate the tools on a 1–5 scale. Scale 3 represented neutral response, and scale 4 and 5 were helpful and very helpful to their learning pharmacology, respectively. Frequency was expressed as a percentage.ResultsTotal 91 responses were collected from 110 students. When combining scale 4 and 5 together which represented the helpful and very helpful, post‐lecture quiz was rated the most helpful (76.9%), followed by making table for antibiotics (72.5%). Practice of baroreflex and volume‐pressure loop were rated positive by 65% of students. TBL was rated fairly positive, with 59.3% at scale 4 and 5, and 54% student liked pre‐lecture quiz, and only 29.9% of students thought the case‐study beneficial.ConclusionsPost lecture quiz, making tables for antibiotics, and practicing of baroreflex and volume‐pressure loop are regarded very helpful to enhance students' learning of Medical Pharmacology.Support or Funding InformationNone
Effects of clavulanic acid on ethanol intake and glial glutamate transporters expression in alcohol preferring rats
Authors: Hakami Alqassem Y. , Sari Youssef 
Published: November 2023

Studies from our lab showed that upregulation of glutamate transporter 1 (GLT‐1) and cystine‐glutamate exchanger (xCT) expression with ceftriaxone, β‐lactam antibiotic, in central reward brain regions was associated with attenuation in ethanol consumption and reinstatement to cocaine‐seeking behavior. In this study, we tested clavulanic acid (CA), non‐antibiotic β‐lactamase inhibitor. This drug has the central β‐lactam pharmacophore, which is very critical for the upregulation of GLT‐1 and xCT expression in the brain. Therefore, we examined the effects of CA on alcohol consumption as well as GLT‐1, xCT and GLAST expression in adult male alcohol‐preferring (P) rats. We found that CA administered at a dose of 5 mg/kg (i.p.) significantly attenuated ethanol consumption in P rats as compared to the vehicle‐treated group. Importantly, we found that CA induced a significant upregulation of GLT‐1 and xCT expression in nucleus accumbens (NAc). There was no effect of CA on GLAST expression in the NAc. These findings revealed that CA at 20‐fold lower dose than ceftriaxone can attenuate ethanol consumption, in part through upregulation of GLT‐1 and xCT in mesolimbic system. Thus, we suggest that CA might be used as an alternative option to ceftriaxone for the treatment of drugs of abuse with hyperglutamatergic state.Support or Funding InformationThis work was supported by the National Institutes on Alcohol Abuse and Alcoholism (Award Number R01AA019458 to Y.S.).
Effects of Self‐Administered Oxycodone on Learning and Cognitive Flexibility in Nonhuman Primates
Authors: Kangas Brian D , Porter Erica N. , Bergman Jack 
Published: November 2023

Over the past decade, prescription opioid abuse has become a major public health concern. Despite increased prevalence, however, the effects of chronic opioid abuse on cognition‐related behavior remains poorly understood. Previous studies in our laboratory have shown that self‐administration of other abused drugs can have adverse effects on cognitive behavior. Therefore, the present studies employed similar techniques to examine the effects of daily intravenous self‐administered oxycodone. Touchscreen‐based repeated acquisition and discrimination reversal tasks were designed to assay basic features relevant to learning and cognitive flexibility, respectively. Four squirrel monkeys were initially trained to self‐administer oxycodone. A range of doses was evaluated in each subject over 1, 2, and 3‐hour sessions until stable intake was observed. A peak dose of 0.1 mg/kg/inj during 3‐hr sessions yielded maximum daily intake and was used for subsequent conditions. Following 30 self‐administration sessions, subjects were then introduced to the touchscreen tasks. Subjects were placed in a touchscreen chamber immediately following oxycodone self‐administration and learned to discriminate between two novel stimuli (acquisition) for a palatable food reward. Once discrimination acquisition was mastered, subjects then re‐learned the discrimination under conditions where the consequences were switched (reversal). Dose‐response functions revealed orderly increases in oxycodone intake as a function of dose. In addition, extending session durations increased daily intake at all doses examined. Stable levels of oxycodone intake using the peak dose of 0.1 mg/kg/inj were maintained over extended periods of self‐administration (>100 sessions). Results from touchscreen sessions indicated dramatic deleterious effects on learning and reversal in some subjects; however, in other subjects less profound effects were observed. Future studies will investigate the effects of abrupt discontinuation of opioid self‐administration (withdrawal) on discrimination learning and reversal.Support or Funding InformationThis research was supported by grant K01‐DA035974 (BDK) from the National Institute on Drug Abuse.
Comparative studies of AMPA receptor positive allosteric modulators PF‐04958242 and PF‐04531686
Authors: Stolz Jacob Robert , Ishii Toshiyuki , Swanson Geoffrey 
Published: November 2023

Fast excitatory transmission in the central nervous system is mediated by α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazolepropionic acid (AMPA) receptors responding to the endogenous neurotransmitter L‐glutamate. Dysfunction of AMPA receptors has been associated with various neurological and neuropsychiatric disorders, but therapeutic targeting of these channels has proved difficult due to the preponderance of side effects associated with AMPA receptor agonists. Positive allosteric modulators (PAMs) of AMPA receptors are thus conceptually attractive therapeutics because they rely on endogenous glutamate signaling to potentiate excitatory signaling. The objective of this work is to study the effects of two PAMs, PF‐04958242 and PF‐04531686, on the biophysical properties of recombinant and native AMPA receptors, with the expectation that the comparative data will prove informative for pro‐cognitive preclinical or clinical studies with these small molecules.We expressed a wide variety of AMPA receptor constructs with and without auxiliary subunits in HEK293T/17 cells, and characterized the effects of PAMs on currents elicited by fast‐application of glutamate. Both PAMs increased glutamate‐evoked current amplitudes and completely eliminated desensitization of all AMPA receptors containing the flip splice isoform, whereas desensitization of currents from flop‐containing receptors was slowed to differing degrees. At saturating PAM concentrations, PF‐04531686 slowed deactivation of GluA1 and GluA4 receptors to a greater degree than PF‐04958242.Comparable effects were seen on deactivation of excitatory synaptic currents (EPSCs) recorded from hippocampal CA1 neurons in slice preparations from wildtype mice. Application (1 mM) of PF‐04531686 slowed both evoked and spontaneous EPSC decays by 3‐fold; in contrast, PF‐04958242 slowed EPSCs 1.5‐fold. At a lower concentration (300 nM), EPSCs were also slowed by PF‐04531686, but not PF‐04958242. There was no effect on paired‐pulse ratio at any concentration of either PAM. These results show that both PAMs slow the decay of EPSCs, resulting from their pharmacological actions on AMPA receptor decay rates. We predict from these data that both PAMs will impact cognition, but that PF‐04958242 might have less risk of unwanted mechanism‐mediated side‐effects due to its more modest modulatory action on synaptic AMPA receptor function.Support or Funding InformationThis project was supported by a contract from Pfizer Neuroscience.