bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mineralocorticoid receptor antagonism prevents associated parenchymal arteriole remodeling
Authors: Yen Ting-Chieh , Laimon-Thomson Erinn , Abolibdeh Bana , Jackson William F. , Dorrance Anne M. 
Published: April 2020

Obesity, a major risk factor for cerebrovascular injury, is associated with an increase in mineralocorticoid receptor (MR) activation. Obesity induced middle cerebral artery remodeling can be prevented by MR antagonism. However, the role of obesity‐induced MR activation in cerebral parenchymal arteriole (PA) remodeling has not been defined. PAs are responsible for regulating microcirculatory blood flow and play a significant role in cerebrovascular resistance. We tested the hypothesis that obesity induced PA remodeling is remediated by MR antagonism. Three‐week‐old Sprague Dawley rats were randomized to a control or high fat (HF) diet. At 12 weeks of age, 50% of rats in each group began treatment with the MR antagonist, Canrenoic acid (20mg/kg/day; CAN). PA structure in 24 week‐old rats was assessed by pressure myography under calcium‐free and zero flow conditions at 60 mmHg. Data are presented as means ± SEM, n = 3–10 per group. MR antagonism increased systolic blood pressure in HF‐diet only (HF+Vehicle: 125 ± 6 vs HF+CAN: 153 ± 6 mmHg, p<0.01). At the time of euthanasia, HF‐fed rats, both vehicle and CAN treated, had more abdominal fat per body weight than controls (Control+Vehicle: 1.44 ± 0.37 vs HF+Vehicle: 4.31 ± 0.44 % of body weight, p<0.01; Control+CAN: 2.26 ± 0.33 vs HF+CAN: 4.70 ± 0.44 % of body weight, p<0.01). HF‐diet decreased PA outer diameter (Control+Vehicle: 85.00 ± 5.51 vs HF+Vehicle: 49.09 ± 3.55 μm, p<0.01) that was reversed by MR antagonism (HF+Vehicle: 49.09 ± 3.55 vs HF+CAN: 96.00 ± 3.49 μm, p<0.01). HF‐diet decreased PA lumen diameter (Control+Vehicle: 68.67 ± 6.74 vs HF+Vehicle: 34.59 ± 2.72 μm, p=0.028) that was reversed by MR antagonism (HF+Vehicle: 34.59 ± 2.72 vs HF+CAN: 73.17 ± 7.38 μm, p<0.01). HF‐diet tended to decrease PA vessel wall area (Control+Vehicle: 1948 ± 55 vs HF+Vehicle: 973 ± 148 μm2, p=0.103); MR antagonism increased wall area in HF‐fed rats (HF+Vehicle: 973 ± 148 vs HF+CAN: 2027 ± 234 μm2, p=0.016). Vascular remodeling is associated with cerebrovascular injury and cognitive impairment. Our results suggest that inward hypotrophic PA remodeling is MR dependent providing a potential therapeutic target to combat obesity‐induced cerebrovascular disease.Support or Funding InformationThis work was supported by National Institutes of Health (NIH) Grants R01‐HL‐137694‐01 to W. F. Jackson and A. M. Dorrance.
O‐GlcNAc Transferase Regulates Growth Factor Signaling and Extracellular Matrix Composition in Idiopathic Pulmonary Fibrosis
Authors: Vang Shia , Helton Eric Scott , Denson Rebecca , Thannickal Victor J. , Krick Stefanie , Barnes Jarrod 
Published: April 2020

RationaleIdiopathic pulmonary fibrosis (IPF) is a chronic disease that affects the lungs. The scarring and progressive fibrosis, mediated by communication from the intracellular network and the extracellular matrix (ECM), results in impaired lung function causing respiratory failure and mortality. Over the years, IPF has been documented as a metabolic and age‐related disease. Several hallmarks of aging have been studied, including dysregulated nutrient sensing, altered metabolism, and intracellular communication. Interestingly, dysregulated nutrient sensing, via altered glucose utilization, has been shown to drive metabolic reprogramming in myofibroblasts. O‐linked N‐Acetylglucosamine (O‐GlcNAc) transferase (OGT) is a metabolic ‘stress sensor’ and is responsible for the O‐GlcNAc modification of proteins involved in many cellular processes including metabolism, intracellular communication, and growth factor signaling. However, the potential role of OGT/O‐GlcNAc on growth factor signaling, intracellular communication, and ECM changes during transdifferentiation has not been studied.MethodsLung tissue from individuals with IPF or controls was subjected to immunohistochemistry (IHC) staining to assess O‐GlcNAc modification. In addition, the role of OGT on transforming growth factor beta 1 (TGFb1) induced fibroblast to myofibroblast transition (FMT) was assessed on primary human lung fibroblasts cultured in the presence or absence of an OGT inhibitor, OSMI‐1. Alpha smooth muscle actin (a‐SMA), type 1 collagen (COL1a1), and Smad3 phosphorylation following TGFb1 stimulation with and without OSMI‐1 was examined by Western blot analysis.ResultsIn human lung tissue sections, O‐GlcNAc levels were increased in IPF lung tissue compared to control human lungs as determined by IHC staining. In primary human lung fibroblast, TGFb1 administration resulted an increase in fibroblast transdifferentiation, which was determined by higher COL1a1 and a‐SMA expression. Interestingly, inhibition of OGT by OSMI‐1 following TGFb1 administration reduced expression of COL1a1 and a‐ SMA. Furthermore, OGT inhibition was shown to alter Smad3 phosphorylation upon treatment with TGFb1 (up to 2 hours) in the human lung fibroblasts.ConclusionCollectively, these data suggest that the metabolic sensor, OGT, and the O‐GlcNAc protein modification may have a role in regulating the expression of ECM proteins including COL1a1 by altering TGFb1 signaling during FMT. More importantly, the OGT/O‐GlcNAc axis may be directly involved in the pathogenesis of IPF and regulation of cellular transdifferentiation through TGFb1 signaling and overall ECM composition. Determining the mechanistic role of the O‐GlcNAc modification of proteins on these processes is needed, and may help to identify novel therapies for the treatment of IPF or any other fibrotic diseases.
Identify effects of phenytoin and losartan on TGF‐beta using human gingival fibroblast‐1 (HGF‐1) cells and establish HGF‐1 cell culture as a model to study drug‐induced gingival hyperplasia
Authors: Lipovskyte Kamile , Burrows Anna , Kale Vijay 
Published: April 2020

PurposeThe main objectives of this study were 1) to evaluate the effects of phenytoin (antiepileptic drug) and losartan (TGF‐beta inhibitor) on cytotoxicity, ATP, and TGF‐beta in human gingival fibroblasts (HGF‐1) cells, 2) to identify whether losartan inhibits gingival overgrowth caused by phenytoin and 3) to establish HGF‐1 cell culture as a model to study druginduced gingival hyperplasia.MethodsHGF‐1 cells obtained from ATCC were grown to 90% confluency in DMEM media supplemented with PenStrep, and bovine serum (FBS 10%). Cells were incubated in 5% CO2, 37°C and 95% relative humidity. For cytotoxicity, LDH leakage in the medium was measured using the CytoTox 96 Non‐radioactive Cytotoxicity Assay Kit, and for 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 phenytoin (0, 10, 25, 50, 100 and 200 μM) and losartan (0, 25, 50, 100, 250 and 500 μM) dissolved in DMSO, and incubated for 24h (30,000 cells/well in 96 well plates), 5% CO2, 37°C and 95% relative humidity. Final DMSO concentration was 0.1%. To identify effects on TGF‐beta, cells were treated with DMSO (0.1% final concentration), Phenytoin (100 μM), Losartan (100 μM), and Phenytoin (100 μM) + Losartan (100 μM). After 24h treatment of cells, TGF beta was measured as per supplier’s protocol using ELISA assay kit (Thermo Fisher, Grand Island, NY). All experiments were carried out as triplicate measurements and mean of triplicates were considered as one experiment (n). Minimum of three repetitions (n = 3) were carried out unless otherwise noted in results.ResultsPreliminary results show that none of the drug concentrations had a cytotoxic effect on HGF‐1 cells. No increase in ATP levels were observed for losartan. However, phenytoin showed greater increase in percent ATP levels when compared to vehicle control. Percent ATP levels for losartan and phenytoin were in the range of 101 ± 4 to 113.5 ± 8.2% and 135 ± 11.1 to 138.1 ± 16.5% respectively. Data from one TGF‐beta experiment showed no increase in TGF‐beta levels with phenytoin treatment, however losartan, and losartan plus phenytoin treatment showed 1.3 and 2‐fold increase in TGF beta levels over untreated cells respectively.ConclusionOur study suggests that phenytoin causes cell proliferation as noted from increased ATP levels. Preliminary results form TGF‐beta experiments were inconclusive and further experiments are needed to ascertain effects of losartan and phenytoin on TGF‐beta in HGF‐1 cells and identify whether HGF‐1 cells can be used as a model to study drug‐induced gingival hyperplasia.Support or Funding InformationThis project is supported by College of Pharmacy, Roseman University of Health Sciences, South Jordan, Utah 84095
Thermal Hyperpnea in Humans Changes Following Passive Heat Acclimation
Authors: Etter Clara Vada , Beaudin Andrew E. , Walsh Michael L. , White Matthew D 
Published: April 2020

IntroductionExercise ventilation was shown to increase after a 10‐day, 2 h·day−1 passive heat acclimation (HA) to 50°C and 20 % RH and this study assessed if thermal hyperpnoea for those at rest show a similar adaptation.PurposeIt was hypothesized that similar to that seen for eccrine sweating and cutaneous blood flow, the core temperature thresholds for human pulmonary ventilation would decrease after heat acclimation. It was also hypothesized that pulmonary ventilation would increase following heat acclimation as it does for an active heat acclimation.MethodsSix male participants performed two 40°C immersions separated by a 10‐day passive heat acclimation. Heat acclimation included 10 consecutive days with participants’ rectal temperatures maintained between 38.5–39.0°C. The heat acclimation was performed in a climatic chamber at 50°C and 20 % RH where participants remained seated at rest for 2 hr·day−1. A repeated measures ANOVA was employed with factors of Acclimation State (Pre & Post) and delta TES (0 to 1.4°C from rest). The p value was set at 0.05. An ANOVA with oxygen consumption as the covariate was employed to assess for heat acclimation‐induced changes in pulmonary ventilation. This study was approved by the SFU Office of Research Ethics.ResultsAcclimation was confirmed by a decrease in resting esophageal temperature (TES) from 37.70 ± 0.19 (mean ± SD) to 37.31 ± 0.11°C (p=0.001), as well as by significant decreases in the TES thresholds for temple cutaneous blood velocity from 37.30 ± 0.12 to 37.07 ± 0.13°C (p=0.014) and for forehead eccrine sweating from 37.60 ± 0.25 to 37.25 ± 0.22°C (p=0.03). The pulmonary ventilation thresholds were 38.28 ± 0.28 before and 37.84 ± 0.44°C after heat acclimation (p=0.13). After accounting for the covariate of oxygen consumption, a significant main effect of Acclimation State (F=10.05, p=0.025) plus an acclimation state x delta TES interaction for pulmonary ventilation (F=3.6, p=0.008) were explained by a significantly greater pulmonary ventilation following acclimation at delta TES of 0.6, 0.8, 1.0°C (0.018 <p < 0.05).ConclusionFollowing a passive heat acclimation the resting TES and TES thresholds for human thermoregulatory responses were significantly lower and pulmonary ventilation was significantly increased during passive heating as indicative of an increased thermal hyperpnea.Support or Funding Information Funding SourceThis project was supported by Natural Sciences and Engineering Research Council of Canada and the Canada Foundation for Innovation.
Novel Mutations in OCTN2 Identified in Newborn Screening Exhibit Reduced Function and Lead to Carnitine Transporter Deficiency
Authors: Koleske Megan , Giacomini Kathleen 
Published: April 2020

Newborn screening programs have revolutionized the diagnosis and treatment of genetic disorders. A compelling example is found in SLC22A5 encoding OCTN2, a bi‐directional plasma membrane transporter, which acts in the influx carnitine and efflux of drugs. Newborns with low plasma carnitine levels and biallelic SLC22A5 variants are diagnosed with Carnitine Transporter Deficiency (CTD), a potentially fatal disorder that can be treated with L‐carnitine supplementation. Here, we sought to validate the role of novel OCTN2 variants in CTD via functional characterization of mutant transporters in vitro.In a recent study (PMID 31364285), three novel OCTN2 missense variants were identified in newborn screening. Following site‐directed mutagenesis and transient transfection in HEK293T cells, we performed isotopic uptake assays using 14C‐carnitine. Sodium phenylbutyrate, a chemical chaperone previously shown to increase function of other OCTN2 variants, was used to treat transfected cells. Welch’s two‐sample t‐test was used to determine significance.Two novel missense variants, p.Asp165Glu and p.Gly473Val, exhibited substantial reduction of carnitine transport, retaining less than 5% of wildtype transporter function (p = 9.63*10−6 and 1.02*10−5, respectively). In contrast, only moderate reduction was observed for p.Met433Thr, which retained 61% wildtype function (p = 0.021). Sodium phenylbutyrate showed trends (which were not significant) toward increasing the function of all three variants, suggesting that intracellular errors of protein folding, processing, or trafficking may contribute to their reduced function.Our studies validate the role of three novel variants in CTD, none of which have been associated previously with the disorder. The studies are consistent with the idea that even partial loss‐of‐function alleles may be disease‐causing for CTD. Finally, we suggest that sodium phenylbutyrate represents a potential modulator of OCTN2 function, with further studies warranted to confirm effects on function and membrane localization of OCTN2 variants.Support or Funding InformationThis work is supported by the Chan Zuckerberg Biohub.
A Meta Analysis of Advancements in Prevention and Therapy of Chronic Kidney Disease
Authors: Negere Mulunesh Lee , Einstein George P. , Tulp Orien Lee 
Published: April 2020

This meta‐analysis study was focused on estimating the prevalence of chronic kidney disease (CKD) in the USA and Ethiopia to show the relevance of global awareness toward chronic kidney disease, advanced treatment practices, and its eradication in these two Nations. Kidney disease has become a global burden, especially CKD, according to CDC 2019 data. CKD is most common in adults. Evidence shows nine in ten adults with chronic kidney disease do not know if they have the disease or not. The estimated ratio shows from the CDC data indicate that 15% of US adults have CKD. Studies indicate that Diabetes mellitus and hypertension are the leading causes of chronic kidney disease more than other factors or diseases. The accepted common treatments for CKD are medication, dialysis, and transplant. The other modality of treatment now emerging is stem cell therapy, which has been proposed to improve the quality of patent life, replace dialysis, and decrease the frustration behind the transplant donor waiting list. In the present study, an extensive search was done with the use of MedLine, EmBase, PubMed, BIOMED Central, CDC Database, Web of Science, and Scopus databases to identify applicable published studies. Search items included CKD prevalence, diabetic and hypertensive causes of CKD, and other common causes of decreased renal function, advancements in CKD treatment, stem cell therapy, clinical trials, diagnostic criterion to diagnose CKD, including possible blood and urine tests, chronic kidney failure, and Renal insufficiency, including pool risk estimates, fixed and random effects models where applicable. The participants were chosen with inclusive criteria using meta‐analysis. The study group was focused on the participants with both diabetic and hypertensive contributors. The results show there is a high prevalence of CKD among diabetes, and the rates for hypertensive patients were consistently high regardless of the country and/or their socio‐economic status. CKD was significantly associated with older age; prevention of Diabetes mellitus and hypertension is mandatory in programs to reduce or eliminate the prevalence of CKD. While stem cell therapy shows promise to decrease the incidence of kidney transplant rate or as a replacement for dialysis, it has not yet become the universally accepted treatment of choice.Support or Funding InformationSupport from University of Science Arts & Technology, Montserrat and the Einstein Medical Institute USA
Evaluation of CB1 antagonists as antidotes for synthetic cannabinoid toxicity in male and female mice
Authors: Raghav Jimit G. , Van Kralingen Thomas , Kumar Hritik , Makriyannis Alexandros , Desai Rajeev I. , Vemuri Kiran 
Published: April 2020

While a plethora of recreational marijuana and cannabis derived products have been legalized across the United States, recent trends also indicate an increase in the number of emergency calls relating to the overdose or accidental exposure to Δ9‐tetrahydrocannabinol (THC) and/or synthetic psychoactive cannabinoids. There are currently no FDA approved medications for treating acute cannabinoid toxicity. To address this public health issue, here we provide key data towards the discovery and development of an effective antidote for treating acute toxicity symptoms that may be associated with cannabis overdose, and by ingestion of certain cannabimimetics such as the potent CB1 receptor agonist JWH‐018. Previously, we established proof‐of‐concept and presented preliminary data showing that the novel CB1 antagonist AM11503 (10 mg/kg, i.v.) effectively reversed the hypothermic effects of an acute dose of JWH‐018 (6 mg/kg, i.p.), effects that were similar to the known CB1 antagonist rimonabant (10 mg/kg, i.v.). Herein, we extend this line of research in male and female CD‐1 mice by: a) examining whether AM11503 induces rimonabant‐like effects on observable behavior; b) assessing the ability of AM11503 to prevent JWH‐018‐mediated tremors and convulsions; and c) comparing the ability of AM11503 and rimonabant to precipitate somatic signs of withdrawal in JWH‐018‐dependent mice. Results show that rimonabant (3.2–32 mg/kg, i.v.) produced a dose‐related increase in scratching and grooming in both male and female mice compared to controls, whereas AM11503 (3.2–32 mg/kg, i.v.) did not alter stereotypic behavior. In interaction studies, administration of 10 mg/kg (i.v.) AM11503 fully blocked tremors and convulsions induced by “suprapharmacological” doses of JWH‐018 (18 mg/kg, i.p.). Finally, rimonabant (10 mg/kg, i.v.) induced significant somatic signs of withdrawal, i.e., head twitches and paw tremors, in male and female mice that were repeatedly exposed to JWH‐018 (3 mg/kg, i.p. once per day for 6 days) compared to vehicle‐treated subjects. In contrast, AM11503 (10 mg/kg, i.v.) did not induce somatic signs of withdrawal in JWH‐018‐dependent mice and effects were similar to vehicle‐treated mice. In conjunction, the inability of AM11503 to produce rimonabant‐like stereotypic behaviors and precipitate somatic signs of withdrawal yet prevent CB1 agonist‐induced acute toxicity allows for the identification and development of safer CB1 antagonist‐based therapies for nullifying synthetic cannabinoid toxicity.Support or Funding InformationFunding Information: Work supported by NIDA/NIH grants R21DA045882 and R41DA044048
Interactions of Pesticides with Drug Efflux Transporter P‐glycoprotein in the European Honeybee
Authors: Pouv Amara , Nicklisch Sascha 
Published: April 2020

Modern agriculture depends on pollinators such as the European Honeybee, Apis mellifera, to produce crops. It is estimated that almost $20 Billion a year in US agricultural production can be contributed to honeybees. However, global bee populations have been in recent decline and the high use of agricultural pesticides have been suggested to directly or indirectly affect bee colony health. Yet, little is known about the types and levels of pesticides accumulation in bees and the underlying mechanism of uptake and distribution. Multidrug resistance proteins from the ABC family, such as P‐glycoprotein (P‐gp), are a first line of defense in biological barriers and responsible for the efflux of drugs and toxins in all organisms.This study aims to determine the molecular interactions of certain prevalent pesticides with the honeybee P‐glycoprotein to predict the bioaccumulation potential of these compounds, including neonicotinoids, glyphosate and fipronil. We are currently cloning honeybee P‐gp from forager bee specimen. Next, we will express and purify honeybee P‐gp and use an optimized ATPase activity assay to define inhibitors, activators and non‐interacting compounds. Compounds identified as P‐gp inhibitors or non‐interacting chemicals will be infused into sugar water and fed to living bees. Total body burden of these compounds will then be determined using LC‐ and GC mass spectrometry. Finally, we will dissect bees and separate the digestive system (foregut, midgut, hindgut) to validate P‐gp expression using Western immunoblotting. Our preliminary data has shown that a commercially available human P‐gp epitope antibody can recognize honeybee P‐gp.Future experiments will aim at studying behavioral changes upon P‐gp inhibitory pesticide exposure, including changes in flight paths and waggle dance in living colonies. In addition, we will determine the interactions of these pesticides with another key efflux transporter in bees, the multidrug resistance protein 1 or MRP1.The results of this study will help to identify the bioaccumulation potential of currently used agricultural pesticides and guide the rational design and use of compounds that are better eliminated by bees. Finally, it will inform farmers, stakeholders and governmental agencies about non‐lethal combinations of pesticides often used as mixtures for proper crop protection.Support or Funding InformationThis study is supported by the Principle Investigator Start‐up funds from UC Davis.