bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of the Dopamine D4 Receptor by Angiotensin II in Rat Astrocytes
Authors: O'Connor Ann Tenneil , Trivedi Malav , Clark Michelle
Published: November 2023

Objective

We investigated whether Angiotensin (Ang) II regulates the Dopamine D4 receptor (D4) protein expression and whether this interaction is dysregulated in astrocytes isolated from Spontaneously hypertensive rats (SHRs).

Background

A role for the cerebellum in attention and cognitive control has recently been reported. Dysregulation of the D4 receptor function in the cerebellum has been linked to neurodevelopmental disorders such as Attention deficit hyperactivity disorder (ADHD). The mechanisms underlying the dysfunction of the D4 receptor are unknown. We proposed that the angiotensin type 1 receptor (AT1R) cross talks with the D4 receptors, and this interaction may be dysregulated in neurodevelopmental disorders such as ADHD. We used the SHR which is a widely accepted animal model for ADHD. Astrocytes were used as a model brain system to explore the interaction between the two receptors.

Methods

Cerebellar astrocytes were prepared from 2–3 days old Wistar and SHR pups. Quiescent cells were treated with 100nM Ang II (the major AT1R ligand) for 1–48 hours. Expression of the D4 receptor protein levels was determined by Western blotting.

Results

There were no significant differences in basal protein levels of the D4 receptor in cerebellar SHR samples as compared to the Wistar controls. In Wistar rats, Ang II downregulated the D4 receptor expression relative to untreated controls. In SHR, Ang II decreased D4 receptor protein expression at most time points examined. There was no significant difference in the ability of Ang II to reduce the D4 receptor protein levels in SHR cerebellar astrocytes as compared to the effect observed in Wistar cerebellum astrocytes.

Conclusion

There is an interaction between the AT1R and the D4 receptor as demonstrated by the ability of Ang II to reduce the D4 receptor protein expression in both SHR and Wistar rat cerebellum samples. This Ang II‐mediated effect was similar in cerebellum astrocytes from both Wistar and SHRs suggesting that these cells were similarly responsive to the peptide. This interaction between the AT1R and the D4 receptor will be further investigated as a potential therapeutic target for pathophysiological conditions mediated by these two systems in other areas of the brain where there is a higher expression of the D4 receptor, such as the prefrontal cortex.
Impact of Delta‐9‐tetrahydrocannabinol on the Reinforcing Effects of Remifentanil in Rhesus Monkeys Responding Under a Food/Drug Choice Procedure
Authors: Maguire David R , France Charles
Published: November 2023

Cannabinoid receptor agonists such as delta‐9‐tetrahydrocannabinol (THC) enhance some (e.g., antinociceptive) but not other effects of mu opioid receptor agonists such as morphine, indicating that opioid/cannabinoid mixtures might be a safe and effective treatment for pain. In rhesus monkeys, cannabinoids fail to enhance and often decrease self‐administration of the mu opioid receptor agonist heroin, suggesting that opioid/cannabinoid mixtures do not have greater reinforcing effects (abuse potential) as compared to opioids alone. Previous studies on the self‐administration of opioid/cannabinoid mixtures in monkeys used single‐response procedures which do not easily differentiate changes in reinforcing effects from other (e.g., generalized rate decreasing) effects, so the interaction between opioids and cannabinoids with regard to reinforcing effects remains unclear. This study used a choice procedure to examine the reinforcing effects of opioid/cannabinoid mixtures relative to food. Rhesus monkeys (n=3) responded under a concurrent schedule wherein responding on one lever delivered 3 sucrose pellets and responding on the other lever delivered i.v. infusions of the mu opioid receptor agonist remifentanil (0.032–1.0 μg/kg/infusion) alone or in combination with THC (32 or 100 μg/kg/infusion). Each session comprised 4 sample trials (2 per reinforcer) followed by 20 choice trials; trials were separated by 10 min. When saline or small doses of remifentanil alone were available, monkeys responded predominantly for food. Responding for drug infusions increased, and responding for food decreased, as the dose of remifentanil increased. For two monkeys, THC did not appear to impact the relative reinforcing effects of remifentanil as the dose‐effect curves for the remifentanil/THC mixtures did not differ from the curves for remifentanil alone. However, for a third monkey, the dose‐effect curve for the remifentanil/THC mixture was shifted leftward compared with the curve for remifentanil alone, suggesting that THC increased the reinforcing potency of remifentanil. In all monkeys, THC (particularly the larger dose of 100 μg/kg/infusion) initially decreased the number of trials completed but that effect dissipated across sessions, suggesting that tolerance developed to rate‐decreasing effects of THC. Overall, these results extend previous studies and show that cannabinoids do not substantially enhance the reinforcing effects of mu opioid receptor agonists and demonstrate the utility of choice procedures for studying the reinforcing effects of drug mixtures.

Support or Funding Information

Supported by the US National Institutes of Health (R01DA005018) and the Welch Foundation (AQ‐0039).
A Scientometric Analysis of Research on Pueraria lobata and Predictable Usefulness in Skin Health
Authors: Sohn Eunsoo , Sohn Eun‐Hwa
Published: November 2023

Pueraria lobata (Leguminosae) has been widely used as a traditional medicine for treating various diseases in Asia. The inclusion of botanical extracts in skin care products is becoming more popular. Potential antioxidant and anti‐inflammatory effects from the natural plant may prove beneficial for the health, texture, and integrity of the skin. The aim of this study is to analyze the global scientific research and estimate its R&D trend in the field of herbs related research through scientometric methods. Using data from the Web of Science database between 2006 and 2015, a total of 1,354 publications was returned by searching P. lobate, scientific name. Knowledge Matrix Plus and VOS viewer software have also been used for the analysis. Results showed that the majority of keywords in the publication papers on the medicinal activities of P. lobata were related to the cardiovascular disease, estrogenic effects, anti‐inflammatory effects, and apoptosis with focusing on the active components, isoflavonoid such as daidzein and genistein. It might also be worth considering that the estrogenic and anti‐inflammatory effects of P. lobate on skin aging and health in the flow‐up study and experiments.
Indole‐2‐carboxamide derivatives as G‐protein coupled Estrogen (GPER/GPR30) ligands
Authors: O'Dea Austin , DeLeon Chelsea , Wilhelm McKenna , Sweeny Patrick , Arnatt Chris
Published: November 2023

Classical estrogen receptors (estrogen receptors α and β), have been shown to cause cell proliferation in a variety of breast cancers upon interaction with estrogenic compounds; however, G‐protein estrogen receptor (GPER) has also been linked to cell proliferation, when in the presence of estrogens in breast cancers absent Estrogen receptors α and β. Here we demonstrate the ability of small molecules, specifically computationally modeled hits that may interact with GPER, to inhibit estrogen binding to GPER. Interactions of these modeled hits were studied on breast cancer cell lines SKBR3, MCF7, and MDA‐MB‐231's in an estrogen promoted antiproliferation assays to search for potential activity with the GPER to determine estrogenic interactions upon the breast cancer lines. These interactions may signify that inhibition of GPER to block estrogen is an effective method for curbing estrogen promoted cancer cell proliferation. Specifically, within MBA‐MD‐231 cells, compounds showed inhibition of growth similar to the GPER selective agonist G‐1, which is consistent with previously published results.
Synthesis and Biological Evaluation of a Novel Series of G Protein‐Coupled Estrogen Receptor Antagonists for The Treatment of Gallstone Disease
Authors: DeLeon Chlelsea , Wilhelm McKenna , Sweeney Patrick , Arnatt Christopher
Published: November 2023

The prevalence of gallstone disease is a major public health issue globally. While laparoscopic cholecystectomy is the current standard of treatment, new research suggests that medicinal therapy may offer a non‐surgical option for patients with a functioning gallbladder. The G protein‐coupled estrogen receptor (GPER) is known to induce gallstone formation in response to 17b‐Estradiol (E2) treatment. Therefore, the design of GPER antagonist may significantly reduce gallstone formation and enable non‐surgical management of gallstone disease. Using a computational homology model of GPER, a novel series of antagonists were designed and synthesized. An original high‐throughput screening (HTS) biochemical assay was designed to analyze GPER activity. Preliminary results show no agonism of intended GPER antagonists.

Support or Funding Information

Saint Louis University
Complete Bilateral Ureteral Duplication: an Incidental Finding in Cadaveric Dissection
Authors: Syed Mohsin , Akhter Noor , Phelan Kevin D , Eltahawy Ehab
Published: November 2023

Ureters are muscular conduits that transport urine from the kidneys to the urinary bladder. During embryonic development, partial or complete duplication of the ureter may result from early splitting of the ureteric bud or the presence of a second ureteric bud. Ureteral duplication is described in approximately 1 in 125 (0.8%) people at autopsy (Nation EF: 1944). Ureteral duplication occurs more frequently in females than in males, which is reported to be approximately 1.6:1. Unilateral duplication is 6 times more common than bilateral duplication. Ureteral duplication is generally associated with ectopia, where one ureter may open in the urethra, vagina, vestibule or epididymal region. Ureterocele is also commonly seen with ureteral duplication. A higher occurrence of uretorenal pathology has been recorded with duplication anomalies. Recognition of the possible effects of abnormal ureteral development is essential in determining the management strategies of the patients with urogenital disorders.

We are presenting a case of a middle‐aged Caucasian female cadaver donor with bilateral ureteral duplication. In this case the upper right ureter (25 cm long) drained 3 major calices (~60% of the kidney); the lower right ureter (22 cm long) drained 2 major calices (~40% of the kidney). On the left, both ureters (upper 27cm and lower 25cm long) drained 2 calices each (50% of the kidney). Each set of ureters entered the urinary bladder enclosed in a common fascial sheath and opened separately in the trigone of the urinary bladder. On both sides, the upper ureteric opening was approximately 5 mm below and medial to the lower ureter. On the right side ureteric openings were marked by a prominent intravesicle ureterocele, however no such ureterocele was observed on the left side. No significant enlargement, constriction or polyps were seen in any of the ureters. The size and parenchyma of both kidneys were within normal range from a gross anatomical perspective. The left Kidney (11.2×4.2×3 cm) was larger than the right kidney (9.5×3.8×1.8 cm).There were no sign of renal cysts, hydronephrosis or calculus in either kidney.

Urologists taking care of patients need to be aware of this normal variation. Of particular interest are endoscopic procedures, where a ureter that harbors a stone or tumor can be missed, if the normal non‐affected ureter was accessed instead.
Dynamic Surface Temperature Control Prevents Overcooling with N6‐Cyclohexyladenosine in Rats
Authors: Bailey Isaac R , Laughlin Bernard , Moore Lucille A , Bogren Lori K , Barati Zeinab , Drew Kelly L
Published: November 2023

Cardiac arrest is a leading cause of death in the United States and currently, therapeutic hypothermia, now known as targeted temperature management (TTM), is the only recent treatment modality proven to increase survival rates and reduce morbidity for this condition. Shivering and subsequent metabolic stress however, limits application and benefit of TTM. Stimulating CNS A1 adenosine receptors inhibits shivering and nonshivering thermogenesis in rats and induces a hibernation‐like response in arctic ground squirrels and rats. We found previously that N6‐cyclohexyladenosine (CHA) is more effective at reducing core body temperature (Tb) in dietary restricted (DR) rats than in ad libitum (AL) fed rats. To develop A1AR agonists for TTM in AL fed animals, the current study evaluates the effectiveness of dose, drug and ambient temperature (Ta) on the Tb lowering effects of CHA and capadenoson in freely fed rats. We found that CHA lowered oxygen consumption (VO2) in AL and DR fed rats, consistent with an inhibition of thermogenesis. CHA at 0.5 or 1.0mg/kg in AL rats produced large variations in response at 17.0°C Ta, but after 1.0 mg/kg, IP, all animals cooled to or below a target Tb of 32°C; some animals over‐cooled to as low as 21°C but re‐warmed to 37°C without incident at Ta 23°C. The partial A1AR agonist, capadenoson (1.0 or 2.0mg/kg, IP) produced a consistent response in all animals and decreased Tb to a minimum of 36°C from a mean of 38°C at Ta 17.0°C during the active phase of the circadian cycle. The response to capadenoson was dose‐dependent. To prevent overcooling after CHA we studied continuous IV administration of CHA in combination with dynamic surface temperature control. Results show this approach to be the best at maintaining a desired target Tb. Rigorous optimization of TTM achieved with CHA and dynamic control of conductive cooling will inform future studies aimed at defining the therapeutic benefit of TTM using drugs such as CHA as adjunctive pharmacotherapy.

Support or Funding Information

Research was supported in part by a graduate student fellowship from The Alaska Native Science and Engineering Program to IRB and by NIH R15NS070779, the Alaska Space Grant Program and Alaska INBRE P20GM103395.
Fractional Contribution of UGT2B15 and UGT2B17 in Testosterone Glucuronidation in Human Liver Microsomes and Its Variability
Authors: Zhang Hae Young , Bhatt Deepak Kumar , Prasad Bhagwat
Published: November 2023

Background and Objective

Testosterone has poor and variable oral bioavailability, thus leading to unreliable predictions of its oral pharmacokinetics. Testosterone glucuronidation is the major route of testosterone elimination, and UGT2B15 and UGT2B17 are considered to be the main enzymes responsible. In order to develop a mechanistic physiologically‐based pharmacokinetic (PBPK) model to predict testosterone first‐pass metabolism and its variability, we estimated the fractional contributions of UGT2B15 and UGT2B17 in testosterone hepatic glucuronidation by performing in vitro activity and quantitative proteomic assays in human liver microsomes (HLM) samples genotyped for UGT2B17 deletion.

Methods

HLM samples were isolated from 29 individual liver tissues from University of Washington, School of Pharmacy liver bank and quantified for UGT2B15 and UGT2B17 protein expression using a validated trypsin digestion protocol. Glucuronidation activity assay was performed in HLM (0.1 mg/ml total protein) containing 5 mM MgCl2‐100 mM phosphate buffer at pH 7.4, alamethicin (0.1 mg/ml), testosterone (2 nM to 20 μM), and UDPGA (2.5 mM), for 30‐minutes. The reaction was quenched by acetonitrile and the sample centrifuged. The supernatant was analyzed by LC‐MS/MS to quantify testosterone depletion and glucuronide formation. Based on activity‐expression correlation and UGT2B17 genotype information, fractional contribution of UGT2B15 and UGT2B17 in testosterone glucuronidation was established. Further, we expanded our quantification of UGT2B17 to a larger pool (n=446) to examine correlation with age, gender, and genotype.

Results

The glucuronidation activity was linear for testosterone incubation concentrations ranging from 2 nM to 20 μM. UGT2B17 showed strong protein expression‐activity correlation (r=0.97) while UGT2B15 showed good correlation only in UGT2B17 gene deletion subjects (r=0.72). Fractional contributions of UGT2B17 and UGT2B15 in testosterone glucuronidation were highly variable across individuals, ranging from 0% to 92%, and 8% to 100%, respectively. UGT2B17 protein expression was associated with gender and age (p<0.05).

Discussion

We identified that UGT2B17 is a major contributor in testosterone hepatic glucuronidation in subjects carrying UGT2B17 gene insertion while UGT2B15 protein abundance correlates with the activity in subjects with UGT2B17 gene deletion. We have established methodology to quantitatively characterize individual contributions of each UGT2B isoform in individual HLM samples. Quantification of correlation between UGT2B17 protein abundance and genotype, age, and gender is crucial for developing PBPK model to predict variability in testosterone metabolism. Further, these data will also be critical for predicting outcomes of doping studies and advancing personalized testosterone replacement therapy.

Support or Funding Information

Department of Pharmaceutics, University of Washington, Seattle, WA
Halogen‐π Interactions in the Cytochrome P450 Active Site: Structural Insights into Human P450 2B6 Substrate Selectivity.
Authors: Shah Manish B , Liu Jingbao , Zhang Qinghai , Stout C. David , Halpert James R
Published: November 2023

Numerous CYP2B6 substrates including drugs and environmental chemicals are halogenated. To assess the role of halogen‐π bonds in substrate selectivity and orientation in the active site, structures of four CYP2B6 monoterpenoid complexes were solved by X‐ray crystallography. Bornyl bromide exhibited dual orientations in the active site with the predominant orientation revealing a bromine‐π bond with the Phe‐108 side chain. Bornane demonstrated two orientations with equal occupancy; in both the C2‐atom that bears the bromine in bornyl bromide was displaced by more than 2.5 Å compared with the latter complex. The bromine in myrtenyl bromide π‐bonded with Phe‐297 in CYP2B6, whereas the two major orientations in the active site mutant I114V exhibited bromine‐π interactions with two additional residues, Phe‐108 and Phe‐115. Analysis of existing structures suggests that halogen‐π interactions may be unique to the CYP2B enzymes within CYP family 2 but are also important for CYP3A enzymes.

Support or Funding Information

ES003619 to Dr. James R. Halpert
Effects of troglitazone on bile acid disposition in Bsep KO rats
Authors: Cheng Yaofeng , Chen Shenjue , Freeden Chris , Chen Weiqi , Zhang Yueping , Nelson David , Humphreys William Griffith , Gan Jinpin , Lai Yurong
Published: November 2023

The inhibition of bile salt export pump (BSEP) by troglitazone (TGZ) and its sulfate metabolite (TS) is one of the mechanisms for TGZ‐induced hepatotoxicity. Here, we evaluated the bile acid profiles in wild type (WT) and Bsep knock‐out (KO) rats following repeated oral dose of TGZ (200 mg/kg/day, 7 days). With the remove of 8 base‐pair of nuclear acids, the mRNA levels of Shp, Mrp2, Mrp3 and Sult2A1 were significantly upregulated in KO rats. Following 7 day TGZ treatment, no signs of liver toxicity were observed in WT and KO rats, but Cyp7A1 was significantly increased in both groups. The plasma exposures of TGZ were not different between WT and KO rats, and are similar on Day‐1 and Day‐7. The exposures of TS were 7.6 and 9.3 fold lower in KO rats than in WT on Day‐1 and Day‐7, respectively. However, the formation of TS in liver S9 incubation was similar between WT and KO, indicating that the decreased TS plasma exposures in KO rats could be from increased biliary excretion, rather than altered metabolism. In general, dosing vehicle had minimal effects on bile acid profiles in WT and KO rats, but plasma exposure of individual bile acids demonstrated variable changes in KO rats when compared with WT rats. Many bile acids were increased with the first dose of TGZ in WT, but the increases were partially restored on last dose. In KO rats, alterations of most bile acids were only observed after 7 doses of TGZ. Therefore, rats seem able to regulate bile acid homeostasis through bile acid synthesis and transporter regulation in the presence of Bsep impairment caused by genetic modification or drug inhibition.
Identification of Misregulated Histone Post Translational Modifications in Melanoma
Authors: Davis Lauren E , Byrum Stephanie D , Mackintosh Samuel G , Shalin Sara , Tackett Alan J
Published: November 2023

Histone post‐translational modifications (PTMs) are involved in the regulation of multiple cellular processes including replication, gene transcription, DNA damage repair and cell cycle progression. Misregulation of histone PTM levels can contribute to diseases and cancers such as melanoma. Melanoma is a tumor arising from the pigment producing melanocytes in the skin. It is the deadliest form of skin cancer, accounting for 75% of skin cancer deaths. In early stages, melanoma is easily treatable with surgery alone, but once it has metastasized it becomes more difficult to treat and survival rates drop. The low survival rate in metastatic melanoma is due, in part, to drug resistance in later stages; therefore, new drug targets need to be identified. The objective of the current work is to identify histone PTMs that are misregulated in melanoma. High resolution mass spectrometry data indicated that histone 4 lysine 20 monomethylation (H4K20me) is misregulated by three fold in malignant melanoma cell lines compared to benign cell lines. H4K20me levels are regulated by the methyltransferase SET8 and the demethylase PHF8, both of which have been reported to play a role in cancer progression. To confirm H4K20me misregulation, immunohistochemistry staining of FFPE melanoma patient samples was performed, which showed that H4K20me was significantly down regulated in malignant tumor tissue compared to benign tissue. In order to assess whether H4K20me levels play a role in melanoma progression, invasion and proliferation assays were performed in melanoma cell lines. Cell lines with high H4K20me were not more invasive than the cell lines with low methylation. However, cell lines with low H4K20me had significantly higher levels of proliferation, indicating that H4K20me may be playing a role in cell growth. Our results indicate that down regulation of H4K20me drives proliferation in melanoma cell lines. Future work will focus on determining how H4K20me down regulation affects cellular processes in melanoma.
Quercetin attenuates Sodium fluoride (NaF)‐induced hypertension through reduction in oxidative stress and heat shock proteins (HSP 70)/extracellular signal regulated kinase (ERK) pathways in rats
Authors: Afolabi Jeremiah Moyinoluwa , Oyagbemi Ademola Adetokunbo , Omobowale Temidayo Olutayo , Asenuga Ebunoluwa Racheal , Ajibade Temitayo Olabisi , Adejumobi Olumuyiwa Abiola , Hassan Fasilat Oluwakemi , Adedapo Adeolu Alex , Yakubu Momoh Audu
Published: November 2023

Sodium fluoride (NaF), a component of certain household items is known to cause renal, cardiac and DNA damage in animals and man. The present study investigated the effect of co‐administration of quercetin on the toxic effects of NaF in rats. Forty male Wistar rats of 10 per group were used for this study. Group A was the control group which was given water; Group B was exposed to 300 ppm (300 mg/L) of NaF alone in drinking water for a week. Rats in Groups C and D were exposed to 300 ppm (300 mg/L) of NaF in drinking water for a week and co‐administered orally with quercetin at 50 mg/kg and 100 mg/kg, respectively. Electrocardiogram and blood pressure parameters including the systolic pressure, diastolic pressure and mean arterial pressure were measured prior to sacrifice of the rats. Biochemical assays, histology and immunohistochemistry on kidneys and hearts tissues were also carried out. Results reveal a significant (p<0.05) decrease in nitric oxide (NO) of rats that received NaF alone alongside a significant (p<0.05) increase in markers of oxidative stress, hydrogen peroxide (H2O2), malondialdehyde (MDA), advanced oxidative protein products (AOPPs) and serum myeloperoxidase. Systolic, diastolic and mean arterial pressure also increased (p<0.05) significantly in NaF alone treated rats. Genotoxicity results revealed increased frequency of Micronucleated Polychromatic erythrocytes (MnPCEs) in NaF alone treated rats. Furthermore, administration of NaF led to a reduction in the expressions of Heat Shock Proteins (HSP 70) and Extracellular signal regulated kinase ERK. Co‐administration of quercetin (50 mg/kg & 100 mg/kg) with NaF (300 mg/L), however led to reductions in the markers of oxidative stress, blood pressure and frequency of MnPCEs, improved antioxidant defence system and increased expressions of ERK and HSP 70, respectively.

Support or Funding Information

The authors declare that this work was self funded
Manganese Exposure Induces Neuroinflammation by Impairing Mitochondrial Dynamics in Astrocytes
Authors: Malovic Emir , Sarkar Souvarish , Harischandra Dilshan S. , Ngwa Hillary A. , Jin Huajun , Anantharam Vellareddy , Kanthasamy Arthi , Kanthasamy Anumantha G.
Published: November 2023

Chronic manganese (Mn) exposure induces neurotoxicity, which is characterized by Parkinsonian symptoms resulting from impairment in the extrapyramidal motor system of the basal ganglia. Mitochondrial dysfunction and oxidative stress are considered key pathophysiological features of Mn neurotoxicity. Recent evidence suggests astrocytes as a major target of Mn neurotoxicity since Mn accumulates predominantly in astrocytes. However, the primary mechanisms underlying Mn‐induced astroglial dysfunction and its role in metal neurotoxicity are not completely understood. In this study, we examined the interrelationship between mitochondrial dysfunction and astrocytic inflammation in Mn neurotoxicity. We first evaluated whether Mn exposure alters mitochondrial bioenergetics in cultured astrocytes. Metabolic activity assessed by MTS assay revealed an IC50 of ~ 100 μM Mn at 24 h in both primary mouse astrocytes (PMAs) and the human astrocytic cell line U373s. Furthermore, Mn treatment reduced mitochondrial mass, indicative of impaired mitochondrial function and biogenesis, which is substantiated by the significant reduction in mRNA of mitofusin‐2, a protein that serves as a ubiquitination target for mitophagy. Seahorse analysis of bioenergetics status in Mn‐treated astrocytes revealed that Mn significantly impaired the basal mitochondrial oxygen consumption rate as well as the ATP‐linked respiration rate. The effect of Mn on mitochondrial energy deficits was further supported by a reduction in ATP production. Mn‐exposed PMAs also exhibited a severe quiescent energy phenotype, which was substantiated by the inability of oligomycin to increase the extracellular acidification rate. Since astrocytes regulate immune functions in CNS, we also evaluated whether Mn modulates astrocytic inflammation. Mn exposure in astrocytes not only stimulated the release of proinflammatory cytokines, but also exacerbated the inflammatory response induced by aggregated α‐synuclein. Lastly, the novel mitochondria‐targeted antioxidant, mito‐apocynin, significantly attenuated Mn‐induced inflammatory gene expression in U373s, further supporting the role of mitochondria dysfunction and oxidative stress in mediating astrogliosis. Collectively, we demonstrate for the first time that Mn drives proinflammatory events in astrocytes by impairing mitochondrial bioenergetics.

Support or Funding Information

NIH grants ES026892 and NS088206
Role of Acyl‐CoA Short Chain Synthetase 2 and Acetyl‐CoA in Regulation of Chromatin Modifications and Gene Expression
Authors: Lindahl Anastasia J. , Moffett John R. , Krishnan Jishnu K. S. , Appu Abhilash , Puthillathu Narayanan V. , Krautkramer Kimberly , Dowell James A. , Namboodiri Aryan M. A. , Denu John M.
Published: November 2023

Post‐translational modifications of proteins control many complex biological processes, including genome expression, chromatin dynamics, metabolism, and cell division. Histone protein acetylation and methylation have been extensively linked to the regulation of chromatin remodeling and gene expression. Chromatin modifying enzymes utilize a variety of metabolites as co‐substrates to catalyze their respective chromatin modifications, closely linking chromatin modifications and metabolism. In this study, we have investigated the role of acyl‐CoA short chain synthetase family member 2 (ACSS2), the cytosolic and nuclear Acetyl‐CoA synthetase of the cell, in the regulation of histone and chromatin protein post‐translational modifications as well as gene expression through proteomic and genomic approaches.

Support or Funding Information

AJL was supported by NIH National Research Service Award T32 GM07215
Elucidate the Role of TGF‐beta in Prevention and Treatment of Gingival Hyperplasia Using HGF‐1 Cells
Authors: Kale Vijay , Mathew Sarah , Melnik Mariya , Danh Lisa M
Published: November 2023

Purpose

The purpose of this study was to evaluate the effects of nifedipine (calcium channel blocker), phenytoin (antiepileptic drug), and losartan (TGF‐beta inhibitor) on cytotoxicity and ATP levels in human gingival fibroblasts (HGF‐1) and evaluate whether losartan attenuates gingival overgrowth caused by nifedipine and phenytoin.

Methods

HGF‐1 cells obtained from ATCC were grown to confluency in D‐MEM media supplemented with PenStrep, and bovine serum (FBS 10%) in a CO2 incubator in T‐flasks. For cytotoxicity (LDH leakage in the medium was measured using the CytoTox 96 Non‐radioactive Cytotoxicity Assay Kit) and ATP (Total % ATP was measured using CellTiter‐Glo Luminescent Cell Viability Assay Kit). For ATP and LDH measurements, cells were treated with various concentrations of nifedipine (5, 10, 25, 50 and 100 μM), phenytoin (10, 25, 50, 100 and 200 μM) and losartan (25, 50, 100, 250 and 500 μM) dissolved in DMSO and incubated for 24 h, 72 h and 7 day periods (1×104 cells/well in 96 well plates, 5% CO2, 37 °C and 95% relative humidity).

Results

Results show that none of the drug concentrations had a significant cytotoxic effect on HGF‐1 cells. No significant increase in ATP levels were observed for losartan. At 24h, nifedipine and phenytoin showed significant increase in percent ATP levels when compared to vehicle control. Percent ATP levels for losartan, nifedipine and phenytoin were in range of 101 ± 4 to 113.5 ± 8.2%; 143.1 ± 16.9 to 148.5 ± 20.4% and 135 ± 11.1 to 138.1 ± 16.5% respectively after 24h treatment.

Conclusion

In conclusion, our study suggests that nifeidipine and phenytoin causes cell proliferation as evidenced from increased ATP levels. Concentrations identified from these studies will be used to assess effects of losartan on nifedipine and phenytoin induced gingival overgrowth using various markers involved in gingival overgrowth.

Support or Funding Information

This project is supported by College of Pharmacy, Roseman University of Health Sciences, South Jordan, Utah
Chemical Composition and In vitro Antioxidant Activity Evaluation of Acetone Extract of Crinum jagus bulbs (Liliaceae) Linn
Authors: Shorinwa Olusayo Aderonke , Ebong Omotayo Oluranti , Siminialayi Iyeopu Minakiri , Obianime Atuboyedia Wolfe
Published: November 2023

Objective

Crinum jagus is popularly referred to as bush onions in Nigeria and is used for the treatment of various diseases in traditional medicine. Oxidative stress is a central pathogenic factor for chronic diseases. Fruits and vegetables have been shown to exert a protective effect against oxidative stress. The primary nutrients thought to provide the protection afforded by fruit and vegetables are the antioxidant phytochemical agents. Hence, this study investigated the in‐vitro antioxidant potential and chemical composition of the acetone extract of Crinum jagus bulbs.

Methods

The plant extract was screened for the presence of phytochemicals while the antioxidant activity was evaluated with 2, 2‐diphenyl‐1‐picryl‐hydrazyl radical (DPPH) and nitric oxide free radical scavenging assay. Total antioxidant activity, total phenol and total flavonoid content were determined using a spectrophotometer. Isolation and characterization of the active principles were done using GC‐MS techniques.

Results

Preliminary phytochemical screening of the plant revealed the presence of flavonoids, tannins, saponins, steroids and alkaloids. DPPH radical scavenging activity of the plant extract (IC50 value) was found to be 72μg/ml while that of the reference standard, ascorbic acid was 31μg/ml. Nitric oxide scavenging activity of extract was found to be 55μg/ml while that of ascorbic acid was 35μg/ml. The plant extract was found to have a tannin content of 194.1 ± 5.33mg/g (expressed as ascorbic acid equivalent), total flavonoids content (220 ± 0.19mg/g expressed as quercetin acid equivalent), tannin (259.6 ± 0.58mg/g expressed as tannin equivalent) and phenol content of 136.9 ± 0.29 mg/g gallic acid equivalent. The active principles were found to be bowdensine and demethoxybowdensine.

Conclusion

The potency of the extract is about 40% (two‐fifth) that of ascorbic acid against DPPH whereas potency of the extract is about 60% (two‐third) that of ascorbic acid for nitric acid inhibition. The findings of this study have shown that the plant possesses good anti‐oxidant activity comparable to the standard.

Support or Funding Information

Self funded
Effect of ultrasound on antimicrobial properties of whey protein‐totarol nanogel
Authors: Guo Mingruo , Ma Shuang
Published: November 2023

Totarol is a natural antimicrobial compound extracted from the heartwood of the Totara tree. The effects of whey protein‐totarol nanogel treated with ultrasound on the growth of Staphylococcus aureus (S. aureus) were investigated. The particle size of WPI‐Totarol nanogels was reduced by ultrasound treatment from 31.24 ± 5.31 nm to 24.20 ± 4.02 nm and its distribution was also narrowed by the treatment. The data of viscosity and modulus indicated that the flow behaviors of WPI‐Totarol nanogels seemed to be a Newtonian and exerted a typical viscoelastic fluid at protein content of 15% (w/v). Rheological properties were more insensitive to ultrasonic time. Time‐killing assays, agar diffusion tests, the cell membrane damage analysis and microstructure were also exploited to study the antibacterial properties of WPI‐Totarol nanogel. The minimal inhibitory concentration (MIC) of WPI‐Totarol after ultrasound treatment decreased from 4 to 2 μg/mL compared to that without ultrasound treatment. WPI‐Totarol nanogels treated with ultrasound resulted in a significant (P<0.05) decrease in time‐killing after 24 h. The agar diffusion results showed that the inhibition zones of WPI‐Totarol nanogels were 12 and 36 mm for untreated and treated with ultrasound, respectively. The bacterial membrane damages and the microstructure changes also proved that WPI‐Totarol nanogels treated with ultrasound had strong antibacterial activities against S. aureus and the antibacterial effectiveness enhanced with the increasing of ultrasonic time. These findings suggested that WPI‐Totarol nanogels treated with ultrasound were more effective against S. aureus than untreated ones.

Support or Funding Information

The financial support for this project was provided by the Ministry of Science and Technology of China (Project # 2013BAD18B07)
Cardiovascular risk factors and academic performance in Peruvian medical students
Authors: Fernandez‐Rodríguez Lissett Jeanette , Bardales‐Zuta Victor Hugo , Altamirano‐Sarmiento Dan Orlando , Hilario‐Vargas Julio Santos
Published: November 2023

Introduction

Cardiovascular diseases (CVD) are the leading cause of death and disability worldwide. Cardiovascular risk factors are present in increasingly younger populations and medical students are no exception.

Objectives

To determine if there is a relationship between cardiovascular risk factors and academic performance in medical students enrolled during 2015 at Antenor Orrego Private University (UPAO) in Trujillo, Peru.

Methods

A descriptive, cross‐sectional study was performed on a population of 78 second‐year medical students (74% female, mean age of 18.9±1.5 years). Family history of CVD, medical history, tobacco use, vital functions, body mass index, waist – hip index (WHI), serum glycemia levels, electrocardiogram and lipid profile (ATP III) were evaluated. Academic performance was measured using the grade obtained in the morphophysiology I course.

Results

Cardiovascular risk factors associated (p<0.05) with poor academic performance were pre‐hypertension in women and men and a WHI > 0.8 in women.

Conclusions

Cardiovascular risk factors correlate with academic performance of medical students, which underlines the importance of mitigating such risk factors.

Support or Funding Information

Antenor Orrego Private University (UPAO)
Proteomic Characterization of Epicardial‐Myocardial Signaling Reveals Novel Regulatory Networks Including a Role for NF‐κB in Epicardial EMT
Authors: Li Yanyang , Urban Alexander , Midura Devin , Simon Hans‐Georg , Wang Q. Tian
Published: November 2023

Reciprocal signaling between the epicardium and underlying myocardium not only is essential for heart development, but also plays a critical role during heart regeneration in regeneration‐competent vertebrates. The complex molecular interactions and regulatory networks involved in this communication are not well understood. In this study, we integrated mass spectrometry with bioinformatics to systematically characterize the secretome of embryonic chicken EPDC‐heart explant (EHE) co‐culture. The 150‐protein secretome dataset established here greatly expands the knowledge base of the molecular players involved in epicardial‐myocardial signaling. We identified proteins and pathways that are implicated in epicardial‐myocardial signaling for the first time, as well as new components of pathways that are known to regulate the crosstalk between epicardium and myocardium. The large size of the dataset enabled bioinformatics analysis to deduce functional networks for the regulation of specific biological processes and predicted signal transduction nodes within the networks. We performed functional analysis on one of the predicted nodes, NF‐κB, and demonstrate that NF‐κB activation is an essential step in TGFβ2/PDGFBB‐induced epithelial‐to‐mesenchymal transition. In summary, we have generated a global perspective of epicardial‐myocardial signaling for the first time, and our findings open exciting new avenues for investigating the molecular basis of heart development and regeneration.

Support or Funding Information

This work was funded by the Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community Trust.
Pravastatin&[trade] Prevents Venous Shear Stress Dependent Induction of Aquaporin 1 Protein Expression in Human Umbilical Vein Endothelial Cells in vitro
Authors: Crum Raphael J. , Krane Carissa
Published: November 2023

Background

Aquaporins (AQPs) are transmembrane water channels that facilitate osmotically‐driven transmembrane water flux. AQP1 is abundantly expressed in vascular endothelium, where it has been shown to function in endothelial cell migration, wound healing and cell volume regulation in response to mechanical stimuli. AQP1 protein abundance is enhanced in cultured human umbilical vein endothelial cells (HUVECs) in response to fluid flow shear stress. Statins, a pharmacological class of HMG‐CoA reductase inhibitors, have been shown to suppress the early development of vein graft intimal hyperplasia by improving endothelial cell function, likely by affecting endothelial response to arterial shear stress. Statins have also been shown to differentially regulate the expression of specific AQPs by altering their abundance and/or subcellular localization. Therefore, it is hypothesized that AQP1 may function as a component of the mechanosensory complex in endothelial cells, and is subject to shear‐stress induced gene regulation and modulation of expression by statins.

Methods

HUVECs were cultured in microfluidic chambers under static (0 dynes/cm2) and venous fluid shear stress (6 dynes/cm2) in the presence or absence of 5 μM Pravastatin™. Fluorescent immunocytochemistry was used to detect AQP1 protein expression and subcellular localization. Images were captured using confocal laser scanning microscopy. Mean fluorescent intensity (in arbitrary units) of AQP1 immunofluorescence under each condition was determined using ImageJ software, and normalized to control conditions (time zero, static culture with no Pravastatin™). 25–30 cells from two independent experiments were analyzed for each experimental condition.

Results

AQP1 immunofluorescence increased 1.3‐fold (p<0.05) after 24 hours, and 1.7‐fold (p<0.05) after 48 hours in cells cultured under venous fluid shear stress as compared to the control. This shear stress‐induced expression was blocked by incubation with Pravastatin™ (p>0.05; control vs. venous shear stress with Pravastatin™). Pravastatin™ had no effect on AQP1 immunofluorescence in static cultures (p>0.05 vs. control). These data indicate that Pravastatin™ prevents shear stress dependent induction of AQP1 in cultured HUVECs.

Conclusions

The results presented show that the shear stress dependent induction of AQP1 protein expression is prevented by Pravastatin™. These results further demonstrate a possible tissue‐independent function of statin drugs in regulating the expression of AQPs in general and specifically a potential cardioprotective benefit of statins, independent of their cholesterol‐lowering function, related to the regulation of AQP1 in vascular endothelial cells.

Support or Funding Information

This research was funded by the Schuellein Chair in the Biological Sciences (CMK), University of Dayton Honors Program (RJC), and a 2015 UGSURF Award to RJC.
Bronchodilator Therapy and Survival in Heart Failure Patients
Authors: Uithoven Katelyn E , Snyder Eric M , Olson Thomas P
Published: November 2023

INTRODUCTION

Bronchodilator therapy via β‐agonists has been considered a controversial treatment option for patients with cardiovascular disease, including heart failure (HF), due to previous studies suggesting increased rates of mortality. A major limitation to several previous studies is the lack of similar baseline characteristics between groups; therefore, a matched control comparison is yet to be explored.

PURPOSE

To evaluate the relationship between bronchodilator therapy and survival in HF patients.

METHODS

A retrospective analysis was performed on 96 HF patients who underwent a cardiopulmonary exercise test between the years 1994–2010. Patients were divided into two groups: 48 patients on bronchodilator therapy (BDT) and 48 matched controls (CTL). Patients were matched based on age (yrs), height (cm), weight (kg), body mass index (BMI), New York Heart Association (NYHA) classification, and cardioselective β‐blocker medication during the time of their cardiopulmonary exercise test. Patients were followed for an average of ten years for an endpoint of cardiovascular mortality. An independent samples t‐test was performed comparing BDT and CTL for age (yrs), height (cm), weight (kg), and BMI, to determine comparable baseline characteristics. A Chi‐Squared test compared NYHA classification, cardioselective β‐blocker status, and sex between BDT and CTL. A log rank test was performed to determine differences in survival distributions between BDT and CTL.

RESULTS

There were no significant demographic differences between groups (age: 59.8 vs. 60.3yrs; height: 172.1 vs. 171.9cm; weight: 86.9 vs. 86.2kg; and BMI: 28.9 vs. 28.6kg/m2 for BDT vs. CTL, respectively). NYHA classification (χ2=0.00; p=1.0), cardioselective β‐blocker status (χ2=0.14; p=0.71) and sex (χ2=0.202; p=0.65) were not different between groups. BDT patients demonstrated an event rate of 51.1% compared to 42.6% for CTL patients over the same time period. The survival distribution for BDT compared to CTL was not statistically different, χ2(1) = 1.49, p=0.22.

CONCLUSION

These data suggest that HF patients on bronchodilator therapy demonstrate no significant difference in survival compared to matched HF controls. In contrast to previous work, our data suggests bronchodilator therapy does not elicit increased mortality in these HF patients.
A Regulatory Role for SDF‐1/CXCR4 Axis In Promoting TNF‐mediated Cardiac Damage Associated With Alcoholic Cardiomyopath
Authors: Little Janay , Abad Adam , Peeran Ahmad , Haddad George , Tarzami Sima
Published: November 2023

INTRODUCTION

Alcoholic cardiomyopathy is a form of heart disease caused by alcohol abuse. Tumor necrosis factor alpha (TNF‐α) plays a role in cardiomyopathy, and can lead to cardiomyocyte apoptosis. Our preliminary data suggests that stromal cell‐derived factor 1 (SDF‐1) stimulates TNF‐α mRNA and protein secretion in cardiomyocytes. Since cardiac expression of SDF‐1 is enhanced in inflammatory cardiomyopathy, we have suggested a regulatory role for SDF‐1/CXCR4 axis in promoting TNF‐α mediated cardiac damage associated with cardiomyopathy.

HYPOTHESIS

Our central hypothesis is that increased levels of SDF‐1 correlate directly with production of TNF‐α and the severity of pathological changes observed in the setting of alcoholic cardiomyopathy. Vascular endothelial growth factor (VEGF) is linked to vasculogenesis, which is seen in healthy hearts. The decrease in quantity of VEGF has been seen as a contributor to damage of heart tissues along with the increase in SDF‐1 and TNF‐α; studies have presented this to be true, but has yet to be shown or seen in alcoholic hearts.

METHODS

Littermate adult rats were put on a 3‐months isocaloric Lieber‐Decarli liquid diet with high alcohol (HA: 100mM) levels. Hearts were taken out and concentrations of TNF‐α, SDF‐1, and VEGF were analyzed in protein lysates that were prepared from those hearts. TNF‐α, SDF‐1, and VEGF protein expression was quantified using ELISA assay. To evaluate whether SDF‐1 upregulation mediated TNF‐α expression and subsequent cell death, adult rat cardiomyocytes were freshly isolated and treated with SDF‐1 at different concentrations and apoptosis was assessed using TUNEL at 24hrs post treatment.

RESULTS

Our data suggests that high alcohol consumption is associated with reduced cardiac contractile function. Moreover, in the setting of alcoholic cardiomyopathy, our preliminary results suggests that SDF‐1 can stimulate production of TNF‐α and subsequent apoptosis possibly contributing to the observed pathophysiology. The ELISA reported that the amount of TNF‐α and SDF‐1 increased in the alcoholic tissues compared to the controls; however, there is a trend toward VEGF downregulation in the alcoholic tissues compared to the controls which can further exacerbate the severity of damage. Treatment of isolated adult rat cardiac myocyte with SDF‐1 at higher concentrations (>30nM) upregualted TNF‐α protein expression and induced subsequent apoptosis.

CONCLUSION

Our data suggests a SDF‐1/CXCR4 network regulates TNF‐α production in cardiomyocytes leading to pathological changes observed in alcoholic cardiomyopathy e.g. myocyte apoptosis.

Support or Funding Information

Research report in this publication was supported by the American Physiological Society under grant number NHLBI; R25 HL115473‐01