bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Atypical Antidepressants Mianserin and Mirtazapine Inhibit TNF‐α‐Induced Apoptosis via LPA1 in Mouse HT22 Hippocampal Cells.
Authors: Olianas Maria C , Dedoni Simona , Onali Pierluigi 
Published: November 2023

Several studies have proposed that pro‐inflammatory cytokines, such as TNF‐α, contribute to the pathophysiology of major depression through different mode of actions, including direct impairment of hippocampal neural progenitor cell development and survival. However, little is known on the mechanisms by which drugs effective in treating major depression may counteract the deleterious effect of pro‐inflammatory cytokines on neural cells. We have recently reported that different antidepressants activate the lysophosphatidic acid receptor LPA1 to induce growth factor receptor transactivation, cell proliferation and neuroprotection [1,2]. In the present study we show that in mouse HT22 hippocampal cells, which display a neuroblast‐like phenotype, the atypical antidepressants mianserin and mirtazapine inhibit the apoptotic cell death induced by TNF‐α and that this protective action involves the activation of LPA1. Thus, prolonged exposure of HT22 cells to TNF‐α increased caspases 8, 7 and 3 activities, proteolytic cleavage of poly ADP‐ribose polymerase and DNA fragmentation. Co‐treatment with antidepressants markedly attenuated the activation of the apoptotic cascade induced by TNF‐α. Blockade of the LPA1 receptor with the selective antagonists AM966 and Ro6842262 antagonized the anti‐apoptotic activity of the antidepressants. Analysis of intracellular signalling showed that through activation of LPA1 antidepressants transactivated the fibroblast growth factor receptor and triggered the phosphorylation of ERK1/2 and CREB, two key regulators of neural survival and differentiation. Collectively, the data indicate that in HT22 hippocampal cells LPA1 is a critical mediator of the neuroprotective effects of mianserin and mirtazapine against the pro‐apoptotic action exerted by TNF‐α.Support or Funding InformationSupported by University of Cagliari. Italy
Acetate, the Metabolite of Ethanol, Increases Cytosolic Calcium and mRNA Expression Levels of EGR1 and TNFα in Dopaminergic Like PC12 Cells
Authors: Behnke Jessica E. , Chapp Andrew D. , Driscoll Kyle M. , Shan Zhiying , Chen Qinghui 
Published: November 2023

We have previously reported that acetate activates NMDAR in vivo and in vitro, increases pro‐inflammatory cytokine mRNA expression, and cytosolic calcium levels, all of which may contribute to neurotoxicity. Neural growth factor (NGF) derived PC12 cells are a reasonable model used in substances of abuse and neurodegeneration studies as they are primarily dopaminergic in nature. Neurotransmitter release is governed in part from rises in cytosolic calcium levels which facilitate loading of synaptic vesicles and exocytosis from the synaptic cleft. We hypothesized that acetate may have similar effects in NGF derived PC12 cells which may facilitate increased dopamine release contributing to alcohol dependence. NGF derived PC12 cells exposed to (2 mM) acetate had a significant (p<0.05) increase in cytosolic calcium relative to baseline, measured in real‐time with a fluorescent calcium probe (Fluo‐4AM). Additionally, acetate (2 mM) significantly (p<0.05) upregulated mRNA expression levels of a calcium dependent protein, early growth response 1 (EGR1) at 15 and 30 minutes post acetate treatment, 5 and 6 fold respectively relative to control. EGR1 is an early response protein implicated to alterations in synaptic plasticity and also the immune response. Furthermore, 30 minute treatment with acetate also significantly (p<0.05) upregulated mRNA expression of tumor necrosis factor alpha (TNFα), 2 fold compared to control. This data suggests that physiological circulating levels of acetate (2 mM) following ethanol consumption and metabolism lead to increased cytosolic calcium levels, and mRNA expression levels of EGR1 and TNFα in dopaminergic like PC12 cells. These in combination may contribute to the development of alcohol dependence and alcohol associated neurotoxicity.Support or Funding Information(Funding support: AHA 16PRE27780121, Andrew Chapp, R15‐HL129213 Zhiying Shan; R15‐HL122952, Qinghui Chen).
Intermittent High Fat Diet Exposure Attenuates Acquisition and Maintenance of Alcohol Drinking in Rats
Authors: Vithlani Nisha , Smith Anna M , Dickenson Kanhiya , Davis Jon F , Sirohi Sunil 
Published: November 2023

Alcohol use disorder (AUD) poses a significant socio‐economic burden to modern society and is a chronic relapsing disorder. Chronic alcohol consumption not only impairs the emotional states but also induces malnutrition. Neuroendocrine peptides, which are known to be regulators of nutritional status, can impact the reinforcing properties of alcohol. Therefore, altered neuropeptides as a result of a nutritionally impaired state may impact alcohol consumption in alcoholics. However, nutritional and dietary alternatives are often overlooked in the management of AUD. We have recently reported that six weeks of intermittent exposure to a nutritionally complete high‐fat diet (NCHFD) attenuates alcohol drinking in rats, which has important clinical implications in AUD treatment. At present, it is unclear if NCHFD pre‐exposure duration less than 6 weeks would be effective enough to impact alcohol drinking. In the present study, we assessed the impact of two weeks of intermittent NCHFD exposure on alcohol drinking in male, Long Evans rats. The rats (n=6/group) were grouped based on their body weight, food and water intake and were given intermittent access to NCHFD (24 hrs, twice a week) or normal chow (controls) for two weeks. All rats had ad libitum access to chow and water always and their daily food intake was recorded. Following two weeks of the intermittent NCHFD regimen, 20% ethanol drinking was evaluated using two‐bottle choice alcohol drinking model on the days following NCHFD exposure. A pattern of caloric overconsumption and under consumption was seen in the NCHFD exposed rats on NCHFD and non‐NCHFD days, respectively. Alcohol intake was significantly reduced in the NCHFD group of rats compared to the chow controls. Furthermore, not only preference and acquisition but also maintenance of alcohol intake was attenuated following intermittent NCHFD exposure. No between‐group differences in water or total fluid intakes were observed, suggesting that intermittent NCHFD exposure selectively impacted alcohol intake. As we begin to understand the impact of altering nutritional status on alcohol drinking behavior, the present data provide fundamental insights into the critical components for evaluating such interventions in the management of AUD.Support or Funding InformationThis publication was made possible by funding, in part, from the NIMHD‐RCMI grant number 5G12MD007595 from the National Institute on Minority Health and Health Disparities and the NIGMS‐BUILD grant number 8UL1GM118967 to SS. We thank the XULA vivarium support, especially Ms. Nannette Simoneaux.
Novel MicroScale Solutions for Biophysical Characterization in Drug Discovery and Development
Authors: Heffern Charles 
Published: November 2023

Here we present 2 unique and proprietary technologies for biophysical characterization of biomolecules to facilitate the drug discovery and drug development process:MicroScale Thermophoresis (MST) monitors the movement of molecules through μm‐sized temperature gradients to quantify interactions of any kind of molecule. Recent results from automated high‐throughput primary and secondary screening campaigns by MST will be presented. In a collaboration project with Sanofi R&D, MST identified several hits which were not detected by orthogonal techniques. In addition, MST detected secondary effects such as protein aggregation and denaturation, and could thus be used to reliably rule‐out false positives and measurement artifacts.nanoDSF technology is specifically tailored for thermal and chemical unfolding studies. By monitoring fluorescence shifts of the amino acid tryptophan or tyrosine, unfolding profiles of multidomain proteins, transmembrane proteins or low‐abundant drug targets can be obtained in an entirely label‐free approach. Here, we present buffer and detergent screening campaigns for membrane proteins as well as for antibodies and antibody‐drug conjugates, highlighting the sensitivity and resolution of this method.In addition, the fully‐automated protein stability platform using the novel Prometheus NT.Plex in combination with NT.Robotic Autosampler enables high‐throughput measurements as required in formulation development. Thermal and chemical unfolding data correlate with long‐term turbidity and monomer content over time, showing that the Prometheus NT.Plex can be used to rapidly predict the long‐term stability of biologics within 1 day.
Functional Improvement of Dystrophic Muscle via let7 Site Blocking Oligonucleotide Mediated Utrophin Upregulation
Authors: Mishra Manoj K , Loro Emanuele , Wilton Steve D , Khurana Tejvir S 
Published: November 2023

Duchenne's muscular dystrophy (DMD) is a fatal genetic disease caused by the absence of dystrophin. Utrophin is the autosomal homolog of dystrophin and when overexpressed can compensate for dystrophin and rescue the dystrophic phenotype of the mdx mouse model of DMD. Utrophin is subject to miRNA mediated repression in muscle by several miRNAs including the let‐7c miRNA. Blocking the let‐7c‐utrophin interaction would be predicted to ‘repress the repression’ and increase utrophin expression. We developed and tested the ability of 2′‐O‐Methyl phosphorothioate (2OMePS)‐based utrophin 3′UTR let‐7c site blocking oligonucleotides (SBOs) to upregulate utrophin and improve the dystrophic phenotype in vivo. Blockade of the let‐7c‐utrophin interaction using bi‐weekly intraperitoneal injections of let7c SBOs for 1 month in young mdx mice led to increased utrophin expression along with improved muscle morphology, decreased muscle fibrosis and increased muscle specific strength. The functional improvement of dystrophic muscle achieved by using 2OMePS‐let7c SBOs suggests a novel, utrophin upregulation based therapeutic strategy for DMD.Support or Funding InformationAcknowledgements: MDA, Shire Plc.
Pharmacokinetics of Cefquinome in Healthy and Pasteurella multocida‐Infected Rabbits
Authors: Elazab Sara T , Schrunk Dwayne , Griffith Ronald W , Ensley Steve M , Dell'Anna Giuseppe , Mullin Kathleen , Elsayed Mohamed G , Amer Magdy S , El‐Nabtity Sameh M , Hsu Walter H 
Published: November 2023

The pharmacokinetics of cefquinome were studied in healthy and Pasteurella multocida‐infected rabbits after a single intramuscular injection at 2 mg/kg. 12 female New Zealand white rabbits (2.0–2.5 kg) were used; 6 of them served as controls, and the other 6 were infected; the experiments were done 1–2 days after Pasteurella nasal inoculation when rabbits showed signs of infection. Plasma concentrations of cefquinome were determined using high‐performance liquid chromatography. The values of elimination half‐life (T1/2 λz), area under the curve (AUC), area under the first moment curve (AUMC), and mean residence time were significantly lower in infected rabbits (0.47 h, 4.54 h*μg/mL, 3.63 h*h*μg/mL and 0.8 h) than healthy rabbits (0.72 h, 9.11 h*μg/mL, 9.85 h*h*μg/mL and 1.1 h), whereas total body clearance was significantly higher in infected than healthy rabbits. Therefore, P. multocida infection caused significant changes in some of the pharmacokinetic parameters of cefquinome in rabbits.Support or Funding InformationEgyptian Ministry of Higher EducationIowa State University supplied the tools and support to carry out the experiments.
Lindane (Gamma‐Hexachlorocyclohexane) Exposure Impairs [Ca2+]i‐Mediated Vascular Reactivity
Authors: Yakubu Momoh A , Abraham Nina S , Bessac Bret F 
Published: November 2023

Gama hexacholorocyclohexane (γHCH; lindane), Gammallin is an organochlorine chemical that has been used as an agricultural insecticide as well as a pharmaceutical treatment for lice and scabies. It is reported to be neurotoxic, interfering with GABA neurotransmitter functions and other biological functions from brain to endocrine and cardiovascular systems. Despite being a banned persistent organic pollutant, it's still in use and can leach and contaminate our foods and water. We have investigated the effects and the mechanism(s) of lindane‐induced vascular dysfunctions in rat aorta. Rats treated with low dose of single pesticide Lindane (100 or 300 μmol/kg;1/5LD50; SP) or multiple pesticides (MP) (aldrin, endosulfan, lindane, 4,4‐DDT, and endrin: 1/100, 1/50, and 1/25 of the LD50, MP) orally or dermal application for 2 or 4 weeks showed significantly attenuated vascular relaxation (57 ± 5% to 36 ± 0.8%; ACh 10−3 M), MP attenuated relaxation (57 ± 5% to 19.8 ±5.8% and 19.0 ±4.0% respectively for 1/100 and 1/25 LD50). Paradoxically, 4 weeks SP enhanced relaxation to ACh (60.5 ± 4.6% to 83.5 ± 8.8%). In aorta from dermal group (SP), relaxation to ACh was significantly attenuated (59.4 ± 5.0 to 23.7 ± 6.0%). Vasoconstriction to PE (10−8 ‐10−3 M) were significantly attenuated by 4 weeks' treatment with oral SP or dermal MP reducing the contraction (0.64±0.06 to 0.15 ± 0.02gm or 0.49±0.01 gm; 10−3 M), but not for 2 weeks oral MP for PE‐induced contraction. Further investigation into the mechanism behind the observed vascular dysfunction reveals that in CHO cell line stably expressing M1‐ACh receptors, Lindane diminishes carbachol (10−8 to 10−3 M)‐induced increases in [Ca2+]i mediated by M1‐ACh receptor. Lindane diminishes Carbachol effects only at Carbachol's maximum activation and these results indicate that the vascular dysfunction by the pesticides could be due to disruption of intracellular Ca2+ regulation. Thus, 24‐hour Lindane exposure facilitates IP3‐mediated [Ca2+]i signaling. Month‐long Lindane exposure induces vascular dysfunction possibly via disruption of IP3‐mediated in [Ca2+]i signaling leading to dysregulated vasoresponsiveness to hormones/neurotransmitters contributing to cardiovascular dysfunctions.Support or Funding InformationThis research was supported by the National Heart, Lung, and Blood Institute (Grants HL03674 and HL070669) and by the use of TSU Research infrastructure support provided by grants G12RR003045 and CO6RR012537 awarded by the National Center for Research Resources, National Institutes of Health (NIH). The G12 program is now a part of the National Institute on Minority Health and Health Disparities (NIMHD) and the C06 program is in the Office of Research Infrastructure Programs in the Office of the Director, NIH.
Rosuvastatin therapy for dyslipidemia and other associated parameters in Saudi metabolic syndrome patients
Authors: Rafeeq Misbahuddin , Habib Hamed , Murad Hussam , Gari Mamdouh , Gazzaz Zohair 
Published: November 2023

BackgroundMetabolic syndrome (MS) is a constellation of metabolic irregularities comprising dyslipidemia, hypertension, hyperglycemia, chronic inflammatory, and hypercoagulable state predisposing to diabetes and cardiovascular events. Statins are first‐line drugs to treat the associated atherogenic dyslipidemia.AimEffect of rosuvastatin on dyslipidemia and other abnormalities of MS in Saudi patients was studied.Settings and DesignProspective, open label, randomized clinical study.Materials and MethodsPatients of either sex ≥18 years (n = 153) having MS as per modified National Cholesterol Education Program Adult Treatment Panel III criteria were prescribed rosuvastatin 10 mg OD for 24 weeks. Serum lipids, biochemical, clinical, and anthropometric parameters were studied before and after treatment.Statistical Analysis UsedStatistical Package for Social Sciences version17 was used. Descriptive analysis was used for all variables and documented as mean ± SD. Normality checked by Shapiro–Wilk test, Kurtosis and Skewness Z‐score, and visualization of histograms. Lipid levels and other parameters before and after treatment were evaluated by paired t‐test for parametric data and Wilcoxon signed rank test for nonparametric data. Pretreatment and posttreatment values were correlated by Pearson's correlation coefficient. Multiple regression analysis was performed to see effect of other variables.Results142 patients completed the study. Highly significant reduction was observed in LDL‐C, TC, TG, VLDL‐C, non‐HDL‐C, Lipoprotein(a) and atherosclerotic index with an elevation in HDL‐C. A total of 86% patients reached LDL‐C goal of ≤ 100 mg/dL. Beneficial response was observed on other associated parameters. There was strong correlation between pre‐treatment and post‐treatment values. No significant effect of age, gender, BMI, obesity and baseline cholesterol was observed on mean cholesterol reduction. No serious/severe adverse effects were reported.ConclusionRosuvastatin markedly improved atherogenic dyslipidemia of MS besides beneficial effects on other abnormalities.Support or Funding InformationThis paper contains the results and findings of a research project that is funded by king Abdulaziz city for science and technology (KACST) (Grant no. ARP‐34‐129)
Contribution of Beta‐Arrestin Signaling on Thyroid Hormone‐Induced Cardiac Hypertrophy
Authors: Lino Caroline Antunes , Barreto‐Chaves Maria Luiza Morais 
Published: November 2023

Thyroid hormones (T3) exert multiple effects on cardiovascular system. It is well known that hyperthyroidism promotes the activation of cardiac Renin Angiotensin System and previous studies have described the involvement of Angiotensin II Receptor Type I (AT1R) in T3‐induced cardiac hypertrophy. AT1R is a seven‐transmembrane domain receptor that promotes signaling transduction by G protein‐dependent and independent mechanisms. G‐protein independent mechanisms are mediated by multifunctional proteins called beta‐arrestins (βARRs), which bind and activate ERK, a classic hypertrophic signaling pathway. Herein, we evaluate the contribution of βARR signaling on ERK activation in cardiac hypertrophy induced by T3. All animal experimentation was conducted in accordance with the Ethical Principles in Animal Research set forth Brazilian College of Animal Experimentation and approved by the Institute of Biomedical Sciences of the University of Sao Paulo. Primary cultures of neonatal ventricular cardiomyocytes obtained from Wistar rats (1–3 days) were stimulated with 15nM T3 (Sigma‐Aldrich; USA). T3‐stimulated cardiomyocytes presented an increase on ERK phosphorylation at 5, 10 and 15 minutes of treatment. The inhibition of AT1R using Losartan (1uM; Sigma‐Aldrich; USA) was able to attenuate such activation, demonstrating the contribution of this receptor. The pre‐treatment with a selective inhibitor of ERK, PD98059 (20uM; Sigma‐Aldrich; USA) evidenced the involvement of this signaling in the increase of hypertrophic markers in T3‐stimulated cells. The contribution of βARRs on ERK activation promoted by T3 is still being evaluated. Besides that, the contribution of βARRs isoforms in the increase of mRNA expression levels of skeletal muscle α‐actin was demonstrated by interfering RNA assays, showing the possible contribution of βARRs signaling to hypertrophy. Our results evidence the involvement of AT1R on ERK activation and the contribution of this signaling pathway on T3‐induced cardiac hypertrophy.Support or Funding InformationFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP 2013/16142‐9 and 2016/13896‐0).
Mono(2‐ethylhexyl)phthalate Exposure Alters Electrical Restitution Curve in Isolated Hearts
Authors: Jaimes Rafael , Sherman Meredith , Siegel Bryan , Posnack Nikki 
Published: November 2023

Medical devices are frequently manufactured with polyvinyl chloride (PVC) plastic, in combination with phthalate plasticizers to impart flexibility to otherwise rigid PVC. Since phthalate plasticizers are not covalently bound, these chemicals migrate out of the plastic device and into lipophilic solutions, including blood, parenteral nutrition, and pharmacological agents. As an example, Di(2‐ethylhexyl) phthalate (DEHP) can make up to 80% by weight of flexible medical devices, including: blood storage bags, medical tubing, and cardiopulmonary bypass and ECMO circuitry. Phthalate leaching from these devices presents a significant intravenous route of exposure to patient populations. Our previously published in vitro studies have indicated that phthalate plasticizer exposure inhibits electrical conduction in isolated cardiac cells.ObjectiveTo investigate the impact of phthalate plasticizer exposure on electrical restitution in isolated whole hearts.MethodsAll animal protocols were approved by the Children's National Institutional Animal Care and Use Committee. Adult rat hearts were excised, the aorta cannulated, and transferred to a Langendorff‐perfusion system. Hearts were perfused with oxygenated Krebs‐Heinslett buffer, and electrodes were placed on the epicardial surface. Mechanical contraction was inhibited with 5 uM Blebbistatin, and hearts were loaded with 200 nM RH237 to optically map action potentials. Hearts were subjected to a ventricular pacing protocol, and action potentials were recorded using an Andor iXon camera (> 500 fps). Electrical signals were collected using Iox2 software (EMKA technologies) and analyzed using custom MatLab scripts.ResultsExposure to clinically relevant concentrations of mono(2‐ethylhexyl) phthalate (MEHP), the main metabolite of di(2‐ethylhexyl) phthalate plasticizer, resulted in a steeper monophasic electrical restitution curve (slope = 1.8) compared with biphasic control (slope = 2.1, plateau slope = 0.1). MEHP‐exposed hearts were also more susceptible to alternans at shorter cycle lengths, compared with control.ConclusionThe effect of phthalate plasticizers on cardiac electrical restitution has important clinical implications. Alterations in action potential duration restitution and the presence of electrical alternans are arrhythmogenic factors. Additional studies are necessary to resolve the impact of plastic chemical contaminant exposure in patients, and the impact on cardiac function.Support or Funding InformationThis study was supported by the National Institutes of Health (R00ES023477 to NGP, UL1TR000075, KL2TR000076), Children's Research Institute: Divisions of Cardiology and Cardiac Surgery.
Lipopolysaccharide increases the expression level of the aquaporin‐4 water channel
Authors: Sugimoto Naotoshi , Nishimura Naoki , Ohnishi Norikazu , Matsumoto Takaaki , Kitaura Takashi , Shido Osamu , Yachie Akihiro 
Published: November 2023

Edema often occurs during an infection or inflammation due to the increase of tissue water permeability. The water permeability of cells is defined by the expression levels of aquaporins that are present in the plasma membrane. In this study, we investigated whether lipopolysaccharide (LPS), used to induce inflammation experimentally, alters the expression level of aquaporins in several cells.Primaryglial cells and C2C12 myoblast cells were maintained in Dulbecco's modified Eagle'smedium (DMEM) containing 10% fetal bovine serum (FBS) at 37°C in a 5% CO2incubator. These cells were treated with LPS (10 μg/ml) for 20 min or 36 h, andthen cell lysates were harvested. The phosphorylated NF‐κB, cyclooxygenase‐2 (COX2), and aquaporin 4 (AQP4) expression levels in the cell lysates were analyzed by western blot using phospho‐specific NF‐κB,COX2, and AQP4 antibodies, respectively.LPS exposure led to the phosphorylation of NF‐κB in both glial cells and myoblast cells after 20 min of stimulation. LPS also increased the expression levels of COX2 and AQP4 in both glial and myoblast cells after 36 h of stimulation. An NF‐κB inhibitor attenuates LPS‐induced phosphorylation of NF‐κB, COX2, and AQP4 expressions in cells.These results suggest that NF‐κB signaling plays an important role in the inflammatory response, including the regulation of COX2 and AQP4 expression. It is possible that overexpression of AQP4 results in a rapid and increased influx of osmotic water across the plasma membrane into cells, thereby inducing cells welling and edema.