bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of maltreatment on cortisol and inflammatory analytes
Authors: Wong Kingston E. , Molnar Danielle S. , Maguire Aindriu R. R. , Moore Jessy , O’Leary Deborah D. , MacNeil Adam J. , Wade Terrance J. 
Published: April 2020

Adverse childhood experiences (ACEs) such as maltreatment have been associated with a disease risk phenotype that is characterized by altered regulation of the hypothalamic‐pituitary‐axis (HPA) and elevated pro‐inflammatory cytokines. However, there is evidence that different subtypes of ACEs elicit different physiological profiles. More specifically children with a history of physical abuse have lower cortisol levels whereas children with a history of sexual abuse have elevated cortisol levels compared to those in other maltreatment and non‐abused subgroups. The objective of the present study was to investigate if different maltreatment exposures, specifically physical and sexual abuse, are associated with HPA axis and inflammation by measuring levels of cortisol and inflammatory analytes. This study examines 156 young adults using the pilot data from the Niagara Longitudinal Heart Study (NLHS). Inflammatory analytes were assessed with blood serum. Retrospective chronic cortisol was measured through hair taken from the back of the scalp. Maltreatment was self‐reported using the Childhood Trust Event survey questionnaire. The NLHS study was approved by the Brock University’s Research Ethic Board. All analysis of variance models were not statistically significant. Those who reported sexual abuse had the highest cortisol levels. Those who reported physical abuse had the lowest cortisol levels compared to other subtypes. Interleukin 6 (IL6), C‐reactive protein (CRP), tumor necrosis factor α (TNFα), soluble tumor necrosis factor receptor 1 and 2 (sTNFR1 and sTNFR2) levels were higher among those with physical and sexual exposure compared to other subtypes. Levels of interleukin 6 receptor α (IL6Rα), glycoprotein (gp130), and interferon γ (IFNγ) were similar across all subtypes. The direction of cortisol levels pertaining to the subtypes of ACEs in the present study was consistent with the literature where physical abuse was linked to lower cortisol levels and sexual abuse was related to higher cortisol levels. Both physical and sexual abuse were trending towards statistical significance to some inflammatory analytes but not others. As the current pilot sample is underpowered for the effect sizes of cortisol and inflammatory analytes, the models used in analysis are likely to achieve statistical significance in a larger sample.Support or Funding InformationThe NLHS is funded by the Canadian Institutes of Health Research (CIHR #s 363774, 399332). ARRM is supported by the Ontario Graduate Scholarship (OGS) program.
Translational and Systems‐based Pharmacokinetic/pharmacodynamic Model for Predicting Abemaciclib and Doxorubicin Clinical Interaction in Rb‐positive Triple Negative Breast Cancer
Authors: Fleisher Brett , Lezeau Jovin , Werkman Carolin , Jacobs Brehanna , Ait-Oudhia Sihem 
Published: April 2020

BackgroundTriple‐negative breast cancer (TNBC) is a breast cancer (BC) subtype that lacks the three hallmark BC receptors: estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. The retinoblastoma protein (Rb) is a potential pharmacological target for TNBC therapy. Its over‐expression in TNBC cells is associated with increased resistance to doxorubicin (DOX) treatment in this BC patient population.MethodsThe drug‐drug interaction (DDI) between DOX (0.01–20 μM) and the CDK4/6 inhibitor, abemaciclib (ABE, 1–6 μM), was examined on TNBC cells that are Rb‐positive (MDA‐MB‐231) and Rb‐negative (MDA‐MB‐468) over a time span exposure of 72 hours. The DDI was quantified for all examined drugs’ concentrations by calculating combination indices (CI) using Compusyn software. Temporal changes in %cell viability were measured using CCK‐8 assay. The fold‐change from control (untreated) and from baseline (time=0) of phosphorylated‐Rb proteins (pRb) were measured using a commercially available ELISA kit. An in vitro pharmacodynamic (PD) model was developed by linking pRb dynamics to temporal changes in %cell viability. Clinical pharmacokinetic (PK) models capturing clinical data extracted from the literature were linked to the developed PD model. A clinically scaled‐up PK/PD model was used to simulate the time‐to‐tumor progression (TTP) as defined by the RECIST and WHO criteria for optimized human dosing regimens of both drugs in combination.ResultsThe combination DOX+ABE is synergistic in MDA‐MB‐231 (CI<1) and antagonistic in MDA‐MB‐468 (CI>1). The potencies (maximum inhibitory effects, Imax) for both drugs were fixed to 1. The estimated sensitivities (IC50, concentrations producing 50% of Imax) were 0.565 (DOX) and 3.23 μM (ABE) for MDA‐MB‐231 and 0.121 (DOX) and 2.03 μM (ABE) for MDA‐MB‐468. Model‐based simulations over 60 days treatment period with DOX (75 mg/m2, intravenous, every 21 days) and ABE (200 mg, oral, twice a day) predicted TTP of 0.5 (DOX) and 6.2 days (ABE) shorter compared to control. Paradoxically, TTP for DOX+ABE was 1.1 days shorter than control suggesting DOX antagonizing ABE effects.ConclusionThe developed translational and systems‐based PK/PD model for DOX and ABE combinatorial effects in Rb‐positive TNBC captured well all observed data (in vitro and clinical). While in vitro experiments in MDA‐MB‐231 cells suggest synergistic DDI for DOX and ABE, model‐based predictions of clinical TTP in Rb‐positive TNBC patients suggest a reduced TTP. Further clinical investigations to support model predictions are warranted.Support or Funding InformationThis work was supported by the American Foundation for Pharmaceutical Education (AFPE).
Assessing the epigenetics effects on the expression of the Fabp3 promoter region of Zinc‐Transporter 7‐deficient mice
Authors: Scherpel Victoria Grace , Kirschke Catherine Patricia , Huang Liping 
Published: April 2020

Zinc is an essential trace mineral for life. Zinc transporter 7 (ZNT7) transports zinc from the cytosol to the Golgi to maintain cellular zinc homeostasis. Mice with null‐Znt7 gene expression exhibit low zinc status, low body weight, as well as alterations in glucose and triglyceride homeostasis leading to a reduction in fat storage in adipocytes, and subsequently an increase in fat accumulation in muscle. The increase in muscle‐triglycerides in the Znt7KO mice is accompanied by a surge in the expression of Fabp3. Fatty‐Acid binding proteins (FABPs) belong to a family of proteins involved in intracellular lipid transport and status, yet some functions remain unknown. Fabp3 is found to be abundantly expressed in the heart, but it can also be moderately detected in skeletal muscle. Altered Fabp3 expression has been linked to a higher risk of developing metabolic syndrome in humans, hence this area of research is promising as rates of obesity and obesity‐related diseases continue to rise. DNA methylation has been shown to regulate human Fabp3 mRNA expression. Therefore, we aim to discover the methylation status of the mouse Fabp3 gene in a zinc deficient state using Znt7KO mice and determine a relationship between cellular zinc deficiency and the increased lipid accumulation in muscle, a cause of insulin resistance in Znt7KO male mice. Our preliminary results indicate that the cellular zinc level‐related DNA demethylation mainly occur in the area surround the transcriptional start site of Fabp3.Support or Funding InformationUSDA/ARS 2032‐51000‐005‐00D
Loss of Repressor Activator Protein 1 Precipitates Cardiac Aging in Mice via p53/PPARα Signaling
Authors: CAI YIN , XIA ZHENGYUAN 
Published: April 2020

BackgroundTelomere dysfunction has been linked to many aspects of the aging process. Increasing evidence indicates that telomere dysfunction leads to cardiac mitochondrial defects. Mammalian repressor activator protein 1 (Rap1) is one of the components of shelterin complex and essential for the maintenance of telomere length and structural integrity. Our preliminary work showed that aged Rap1 knockout (Rap1−/−) mice exhibited more pronounced aging‐associated phenotypes (including earlier hair graying, hair loss, reduced body weight). However, the effects of Rap1 on mitochondrial function and its contribution to cardiac aging are largely unknown. Thus, the present study investigated whether or not loss of Rap1 in mice precipitates cardiac aging and explored the underlying mechanisms.Methods and ResultsMice cardiac function was assessed with transthoracic echocardiography performed noninvasively using a Vevo 2100 high‐resolution imaging system. Deletion of Rap1 in mice aggravated aging‐related cardiac structural changes and dysfunction, as evidenced by increased left ventricular (LV) posterior wall end‐diastole and LV mass, reductions in ejection fraction and fractional shortening, as well as significantly reduced myocardial performance index (all p < 0.05, Rap1−/− vs. age‐matched counterparts). These changes were associated with greater cardiac senescence (enhanced SA‐β‐gal staining), cardiac hypertrophy (enhanced wheat germ agglutinin staining and increased the ratio of heart weight with body weight), abnormalities in the mitochondrial ultrastructure (cristae fragmentation, vacuolization). Mechanistically, Rap1 deficiency led to shorter telomere (T/S ratio), enhanced DNA damage (γH2AX) and increased nuclear p53 level in vivo in the myocardium and in primary cardiomyocytes when compared to age‐matched counterparts (one year old). Chromatin immunoprecipitation revealed that p53 directly binded to the promote site of PPARα [a central regulator of cardiac fatty acid metabolism (FAM)] and repressed its expression. Rap1 deficiency impaired FAM (reduced CD36, CPT1α, ACADL level and palmitate‐induced oxygen consumption), while PFTα (a p53 inhibitor, i.p. 1.1 mg/kg/day) treatment in Rap1−/− mice significantly alleviated cardiac aging and enhanced FAM by restoration of PPARα.ConclusionsWith aging, Rap1 deficiency may lead to shorter telomere length, increase DNA damage and thereby activate p53, which in turn suppresses PPARα, leading to impaired FAM and mitochondrial defects with abnormal cardiac structure and dysfunction. These findings may identify a new cardiac dimension in the physiological role played by telomere‐Rap1.Support or Funding InformationThe authors’ work was supported by the General Research Fund (17123718M, Research Grants Council of Hong Kong).
IEX‐1 is Required for Vitamin D‐induced Inhibition of Proinflammatory State of Macrophages
Authors: Khan Faaeiza , Owens Elan , Singh Sandeep , Ahern Alex , Shahid Mohd 
Published: April 2020

Vitamin D (Vit‐D) deficiency is linked to several metabolic diseases. However, the underlying mechanism remains elusive. Immediate early response gene x‐1 (IEX‐1)—which is negatively regulated by Vit‐D—plays a pathogenic role in obesity and atherosclerosis. IEX‐1 paucity inhibits macrophage‐mediated inflammation and protects against these diseases. We hypothesize that Vit‐D promotes anti‐inflammatory state of macrophages via inhibiting IEX‐1 expression.We harvested bone marrow cells from wild‐type (WT) and IEX‐1 knockout (KO) mice and cultured them with macrophage‐colony stimulating factor (M‐CSF) to mature into macrophages. The cells were differentiated into pro‐ or anti‐inflammatory macrophages by treatment with LPS (10 ng/ml) or IL‐4 (10 nM), for 24 hours in the absence/presence of Vit‐D (10 nM). Total RNA was extracted and cDNA was synthesized. The mRNA levels of IEX‐1, pro‐, and anti‐inflammatory marker genes were quantified by qRT‐PCR. The TNF‐α protein level was measured in supernatant using ELISA. Further, impact of IEX‐1 deficiency on LPS‐induced pro‐inflammatory macrophage formation and apoptosis was determined using flow cytometry. Vit‐D inhibited LPS‐induced increases in mRNA levels of TNF‐α (42±5 vs. 58±5‐fold, p<0.05) and iNOS (327±29 vs. 523±102‐fold, p<0.05) in WT cells. Vit‐D also suppressed LPS‐induced TNF‐α protein production by macrophages (18±1 vs. 27±1 ng/ml, p<0.05). Conversely, Vit‐D potentiated IL‐4‐induced increases in expression of anti‐inflammatory genes Arg‐1 (15204±2780 vs. 4362±1624‐fold, p<0.01), MRC‐1 (13±3 vs. 6±1‐fold, p<0.05), and Clec10a (8±1 vs. 4±1‐fold, p<0.05). However, these effects of Vit‐D were markedly attenuated in IEX‐1 KO cells. In WT cells, Vit‐D treatment dose‐dependently decreased IEX‐1 expression, showing negative regulation of IEX‐1 by Vit‐D. Flow cytometry analysis demonstrated that IEX‐1 deficiency alone was sufficient to inhibit LPS‐induced pro‐inflammatory macrophage formation (11±1 vs. 26±3%, p<0.05). This effect was concomitant with an increase in apoptosis (10±1 vs. 18±2‐fold, p<0.05) during LPS treatment.These results suggest that Vit‐D favors the anti‐inflammatory state of macrophages in part via repressing IEX‐1 expression.Support or Funding InformationCTRE Faculty Development Seed Grant, Chicago State University
Ameliorative effects of Moringa oleifera (MO) seeds on L‐NAME‐induced hypertension in rats
Authors: Omóbòwálé Temidayo Olutayo , Oyagbemi Ademola , Aderoju Abiodun , Adejumobi Olumuyiwa A. , Adedapo Adeolu , Gbadamosi Idayat , Yakubu Momoh 
Published: April 2020

Cardiovascular diseases including hypertension, have been recognized as leading causes of death worldwide. In this study we investigated the effect of Moringa oleifera on L‐NAME induced hypertension in rats. Fifty (50) Wistar albino rats were grouped into five groups. Group A, the control, was administered water only. Groups B‐E received orally, 40 mg/kg of L‐NAME only, 40 mg/kg of L‐NAME and 10% MO feed inclusion, 40 mg/kg of L‐NAME and 20% MO feed inclusion, and 40 mg/kg of L‐NAME and 10 mg/kg of lisinopril, respectively for 35 days. Treatment with L‐NAME caused a significant (p< 0.05) reduction in Nitric oxide concentration as well as an increase in blood pressure measurements when compared with the control. These were however ameliorated by the administration of MO seeds and lisinopril. Markers of oxidative stress (malondialdehyde and Hydrogen peroxide) increased significantly (p<0.05) in the group treated with L‐NAME alone while the administration of MO seeds and lisinopril caused a significant decrease in their values when compared with the controls. Also, the administration of L‐NAME caused a significant (p<0.05) decrease in the activities of enzymic antioxidants (Glutathione peroxidase and Glutathione‐S‐transferase) while there was reduction in the levels of reduced glutathione level as well as activities of superoxide dismutase, glutathione peroxidase and glutathione‐S‐transferase when compared with the control. These were however significantly (p<0.05) ameliorated by the administration of MO seeds and lisinopril. Taken together, Moringa oleifera seed inclusion in the feed of rats ameliorated L‐NAME induced hypertension by improving Nitric oxide bioavailability as well as its antioxidant activitiesSupport or Funding InformationNone
Modulation of Cytokine Secretion from Thp‐1 Macrophages by Adenosine Receptors
Authors: Eudy Brandon James , da Silva Robin P. 
Published: April 2020

Macrophages serve a major role in the innate immune system by directing the inflammatory response in tissues. By secreting cytokines and chemokines, macrophages direct other immune cells to sites of inflammation. Once inflammatory stimuli are removed, macrophages play a major role in initiating resolution of inflammation by secreting anti‐inflammatory cytokines, greatly influencing immune function and chemotaxis. However, the signals which tune cytokine production towards resolution of inflammation are not yet fully understood. Small metabolites such as adenosine can initiate cell signaling pathways which direct cytokine secretion. Adenosine acts on four cell surface g‐protein coupled receptors (A1, A2A, A2B, and A3 adenosine receptors) and is understood to be a potent immunosuppressive molecule by stimulating IL‐10 secretion. However, the full influence of adenosine receptor agonism on cytokine secretion is not known. Therefore, our primary aim was to determine how adenosine alters the pattern of cytokines secreted during an inflammatory response. We used a multiplex immunoassay to measure secretion of several cytokines in conditioned medium from LPS‐stimulated THP‐1 monocyte derived macrophages treated with adenosine or a vehicle control. We also used siRNAs directed against each adenosine receptor to determine which receptor mediated the effect of adenosine on each of these cytokines. LPS‐stimulated THP‐1 macrophages treated for four hours with adenosine showed increased IL‐10 and IL‐1β secretion and reduced IL‐6 and MCP‐1 secretion. We found the A1 receptor is required for the stimulatory effect of adenosine on IL‐10 and IL‐1β secretion while the A2B receptor is required for the inhibitory effect of adenosine on IL‐6 and MCP‐1 secretion. We also show that under basal adenosine concentrations the adenosine receptors serve to suppress and modulate various cytokines. These data show adenosine receptors are important regulators of cytokine secretion in macrophages. Adenosine, by signaling through the A1 and A2B adenosine receptors, stimulates IL‐10 and IL‐1β secretion while reducing IL‐6 and MCP‐1 secretion which signals priming of immune cells and tips the balance towards a pro‐resolution state which is of value in the context of several pathologies involving chronic inflammation such as metabolic syndrome, arthritis, and cancer. Future studies should determine how adenosine, by altering cytokine patterns, may affect other aspects of macrophage phenotype including ROS production and phagocytic function.Support or Funding InformationFunded by the UF/IFAS Agricultural Experiment Station.
Regulator of G protein signaling complex, Gβ5‐R7, promotes glucose‐ and extracellular signal‐stimulated insulin secretion
Authors: Slepak Vladlen , Wang Qiang , Henry Taylor , Pronin Alexey , Lubaczeuski Camila , Jang Geeng-Fu , Crabb John , Bernal-Misrachi Ernesto 
Published: April 2020

Glucose‐stimulated insulin secretion (GSIS) is essential for maintaining energy homeostasis. G protein‐coupled receptors (GPCRs) are important modulators of this process by extracellular signals such as hormones, neurotransmitters and non‐glucose metabolites. We investigated the role of Gβ5‐R7, the protein complex consisting of the atypical G protein β subunit Gβ5 and a regulator of G protein signaling (RGS) of the R7 family. To analyze the effects of Gnb5 knockout on insulin secretion we used mouse insulinoma MIN6 cell line and pancreatic islets isolated from the Gnb5−/− mice. Consistent with previous work, Gnb5knockout diminished insulin secretion evoked by cholinergic agonist Oxo‐M. Here, we found that Gnb5 knockout also attenuated activity of other GPCR agonists ADP, AVP, GLP‐1, forskolin and, surprisingly, the response to high glucose. Our experiments also provided evidence that Gnb5 knockout eliminated the positive effect of cell adhesion on Oxo‐M‐stimulated GSIS, likely through adhesion GPCR GPR56. Total insulin content and the amount of insulin released in response to K+‐evoked membrane depolarization in Gnb5 −/− islets or MIN6 cells was normal. The pleiotropic effect on several GPCRs led us to the hypothesis that Gβ5‐R7 may play a non‐canonical role in insulin secretory pathway downstream of G protein signaling and glucose‐activated membrane depolarization. We found that Gnb5 knockout does not influence cortical actin depolymerization, but influences activity of protein kinase C, and thus far identified 14‐3‐3ɛ as one of the affected substrates. Together, these results allow us to propose that the Gβ5‐R7 complex regulates a phosphorylation event participating in the vesicular trafficking pathway, downstream of G protein signaling and actin depolymerization, but upstream of insulin granule release.Support or Funding InformationThis work was supported by National Institutes of Health Grants R01DK1054127(to V.S.), RO1‐DK073716, DK084236 (to E.B.M), EY11373 (to J.C.).
E‐Cigar Vapor Alters Lung Fibroblast Viability via Alpha 7 Nicotine Acetylcholine Receptor
Authors: Nho Richard 
Published: April 2020

RationaleIdiopathic Pulmonary Fibrosis (IPF) is the most lethal fibrotic lung disease characterized by the presence of apoptosis‐resistant lung fibroblasts and relentless production of a collagen‐rich extracellular matrix. Recently, e‐cigars become popular in young adults due to their easy availability, various e‐liquid flavors, and belief that e‐cigars are safer than tobacco products. However, there is a growing concern that the exposure to e‐cigar vapors can increase the risk of developing lung diseases including lung fibrosis. However, the patho‐biological effect of e‐cigar vapor on lung fibroblasts has not been understood.Study objectiveThe objective of this study is to examine the pathological effect of e‐cig vapor on fibrotic and non‐fibrotic lung fibroblasts and to elucidate the underlying mechanism that alters fibroblast viability.MethodsPrimary lung fibroblasts from IPF and non‐IPF patients (n=7, each) were treated with Aroma E‐juice Dark Honey e‐cig aerosol extracts (EAEs) and Marlboro Gold smoke extracts that contain 10 ng/ml of nicotine. Fibroblast viability and proliferation were measured on 3 D collagen matrices. Fibroblasts were then treated with various does of nicotine and nicotine‐free e‐liquid (NicVape Retro Fruit Twist flavor), and their viability and proliferation were measured under the same condition. Molecular assays were carried out to elucidate the cellular mechanism that alters fibroblast viability.ResultsWhen non‐IPF lung fibroblasts were treated with EAEs, their viability was significantly increased compared to that of smoke extract‐treated fibroblasts. Likewise, EAE‐treated IPF fibroblast viability was also highly increased. However, smoke extract treatment had minor effects on their viability. Similar to this finding, non‐IPF and IPF fibroblast proliferation was not changed in response to EAEs. Our study further showed that nicotine in EAEs predominantly alters control and IPF fibroblast viability. In contrast, nicotine free e‐liquid did not affect their viability. Our molecular assays further revealed that α7 nicotine acetylcholine receptor (α7nAChR) expression was up‐regulated in nicotine and EAE‐treated lung fibroblasts. However, the utilization of a selective α7nAChR inhibitor, methyllycaconitine and siRNA significantly sensitized non‐IPF and IPF fibroblasts to collagen matrix‐induced cell death by inhibiting a fibroblast survival pathway.ConclusionsOur results suggest that exposure to e‐cig vapor alters fibroblast viability and nicotine in e‐cigs plays a crucial role in altering a fibroblast phenotype via the α7nAChR‐dependent apoptosis‐resistant pathway. These results show a possibility that the inhalation of e‐cigar vapor is a risk factor to promote lung fibrosis and that the selective inhibition of α7‐dependent cellular pathway may be an effective approach to reduce e‐cig vapor‐induced lung fibrosis.Support or Funding InformationThis study was supported by the University of Minnesota Medical School Faculty Research Award and Center for Lung Science and Health grant.