bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of Lymphatic‐Targeted Fluorescent Organic Nanoparticles to Label Cells in vitro
Authors: Petersen Chase Benson , Morris Kahlen , Svechkarev Denis , Mohs Aaron M. , Fink Darci M. 
Published: June 2021

We have developed a new class of hyaluronic acid (HA)‐conjugated fluorescent organic nanoparticles. These conjugates are designed to specifically target lymphatic endothelial cells (LECs) based on the ligand‐receptor relationship between HA and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE‐1). The lymphatic vasculature undergoes dramatic remodeling during inflammation, wound healing, and in the tumor microenvironment, and is a significant route of metastasis. The long term goal of this project is to develop new tools for pre‐clinical and clinical imaging of pathological lymphatic vessel remodeling signatures that are indicative of the presence of early malignancy. Our work here has focused on 10 kDa and 100 kDa HA‐dye conjugates; future translational experiments will test nanoparticles with a paramagnetic core. The primary objective here was to optimize the dosage for delivery of HA‐conjugated fluorescent dyes to immortalized mouse and primary human LECs for detection by fluorescence microscopy. This included identification of LYVE‐1 expressing experimental cell lines, analysis of cell death, and dose‐dependent HA‐dye in vitro labeling studies. The toxicity of the HA‐conjugated dye was determined using a caspase 3/7 glo luminescence assay that measured the amount of cleaved caspase 3/7 in HA‐treated mouse and human lymphatic cells. The dye was found to be non‐toxic to both the mouse and human lymphatic cells at all dilutions tested including the working staining dilution as compared to staurosporine control used to induce apoptosis. LYVE‐1 expression in human and mouse LECs was confirmed by western blotting and immunofluorescence microscopy techniques including epifluorescence and spinning disk confocal. Fluorescence microscopy analysis showed a dose‐dependent increase in HA‐conjugate dye labeling of LECs. We have shown that HA‐conjugated dyes are non‐toxic to both mouse and primary human LECs, and that this conjugate can experimentally label cells by fluorescence microscopy. Ongoing tubulogenesis and migration experiments are designed to determine if LEC function is impaired with the HA‐conjugate dyes bound. Co‐culture and corneal explant assays will confirm specificity of HA‐conjugate dye for LECs.Support or Funding InformationIn collaboration with Dr. Aaron M. Mohs and Denis Svechkarev at the University of Nebraska Medical Center and bio‐systems networks & translational research (BioSNTR)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Estrogen Inhibits Colon Polyp Formation in a Benzo(a)Pyrene‐Induced Colon Cancer Rat Model
Authors: Harris Kenneth J. , Amos‐Landgraf James , Archibong Anthony E. , Adunyah Samuel E. , Ramesh Aramandla 
Published: June 2021

Colorectal cancer (CRC) is the third most common diagnosed cancer and the third leading cause of cancer‐related deaths in the United States. The incidence and mortality rates of colorectal cancer are higher in men than in women. Epidemiological evidence show estrogen might influence the incidence of CRC in women by acting in a protective role. Whether estrogen acts via estrogen receptor beta (ERβ) to render its protective effect or through some other mechanism is not yet known. Using the Polyposis in Rat Colon (PIRC) animal model, we have demonstrated that male animals develop twice as many polyps in the colon than female animals when treated with the food toxicant, Benzo(a)pyrene [B(a)P]. In this study, to assess the potential protective role of estrogen against polyp formation in the colon, we treated female PIRC rats, depleted of ovarian steroids by ovariectomy (OVX), with B(a)P and measured polyp development. We found that B(a)P treatment of OVX‐female PIRC rats increased the polyp numbers when compared to control OVX‐female animals. Estrogen replacement in these B(a)P‐treated OVX PIRC rats significantly decreased total colon polyp numbers. These results suggest that estrogen does provide protection against B(a)P‐induced colon carcinogenesis.Support or Funding InformationThis research was funded by NIH grants 5R25GM059994‐3, U54CA163069‐04 and G12MD007586‐29.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interactions of N, N‐ diethyl‐meta‐toluamide (DEET) and Novel Insect Repellents with Mammalian GABAA and Glycine Receptors
Authors: Grant Gariel G , Hall Adam C 
Published: June 2021

Mosquito‐borne diseases such as malaria, dengue and yellow fever are responsible for millions of deaths annually. Repellents disrupt interactions between humans and insects, thus providing personal protection from bites through which diseases are transmitted. The most commonly used repellent today is N, N‐ diethyl‐meta‐toluamide, DEET. Despite its popularity, DEET is not effective against all species of mosquitoes, and there are widespread concerns about its possible toxicity to users. Consequently, there is a demand for new repellents which are safer, effective for longer durations, and more efficacious against a broader range of arthropods. The novel repellents tested in the following study fall under two classes of chemicals: acylpiperidines and carboxamides.In the present electrophysiological investigation, the neurotoxicity of these novel repellents was explored relative to that of DEET, focusing on the effects of the repellents on inhibitory neurotransmitter receptors. Two‐electrode voltage clamp technique was employed with Xenopus laevis oocytes expressing mammalian GABAA (α1β3γ2S) and glycine (α1 homomer) receptors. Results indicated that high concentrations of DEET negatively modulated glycine receptor activity when co‐applied with submaximal glycine concentrations. 1 mM DEET inhibited glycine receptor activity by 48.4% ± 4.0% and 10 mM DEET produced a maximal inhibition of 80.4% ± 3.2%. Interestingly, most carboxamides (including DEET) inhibited the activity of glycine receptors, while the acylpiperidines enhanced the activity of glycine receptors, acting as positive modulators. Neither acylpiperidines nor carboxamides were found to inhibit the activity of GABAA receptors, with DEET and other carboxamides acting as weak agonists at high concentrations (>1 mM) to produce directly‐activated GABA currents. The EC50 for the activation of GABAA receptors by DEET was 4760.4 ± 50.6 μM, compared to a GABA EC50 of 42.9 ± 8.0 μM. These data suggest that DEET and the novel repellents have distinct effects on mammalian GABAA and glycine receptors. Furthermore, the results highlighted a stark difference between the two classes of chemicals, whereby acylpiperidines have the potential to result in less neurotoxicity relative to carboxamides. Like strychnine, carboxamides, including DEET, act as competitive antagonists at glycine receptors albeit with considerably less potency.Support or Funding InformationBlakeslee and Holmes funding, Smith CollegeThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Berberine attenuates the acute pancreatitis induced by Dibutyltin dichloride (DBTC) in albino Wistar rats
Authors: Putta Swathi , Qureshi Absar Ahmed , Kilari Eswar Kumar 
Published: June 2021

Acute pancreatitis is a life threatening inflammatory disorder of pancreas. Currently, there is no effective pharmacological therapy available for this disorder and they does not showed any effect on root cause of the disease. The objective of the study was to evaluate the protective role of berberine in Dibutyltin dichloride (DBTC) induced acute pancreatitis in Wistar albino rats. The rats were divided into 4 groups and each carrying 6 rats. Group 1 served as control, Group 2 served as DBTC control (6mg/kg., i.p), Group 3 and 4 received 50mg/kg and 100mg/kg of berberine orally for a period of 28 days. Blood samples were collected at 1st, 3rd day and 28th day from all groups for the estimation of serum amylase and lipase. The DBTC reported to have effect on both exocrine and endocrine portion of the pancreas. The serum blood glucose, liver and kidney function markers (SGOT, SGPT, ALP, bilirubin, creatinine and protein); oxidative stress parameters such as SOD, CAT, GSH, Nitrite, MDA and Protein carbonyl content were estimated in both pancreatic and liver homogenates; inflammatory mediators (IL6, TNF α and CRP) were also estimated in both pancreatic and liver homogenates at the end of 28 days treatment period. The histopathological study was conducted for pancreas and liver. The treatment with berberine showed significant reduction of fasting blood glucose and liver kidney markers in DBTC induced pancreatitis in rats. The antioxidant enzymes SOD, CAT and GSH were found to be elevated and oxidative stress parameters (Nitrite, MDA and Protein carbonyl content) were found to be reduced with treatment with berberine in DBTC induced pancreatitis in rats. The berberine normalizes the elevated inflammatory cytokines with DBTC induction. The histopathology study indicating that berberine ameliorated the necrosis of both pancreatic acini and hepatocytes in DBTC induced pancreatitis in rats. The possible protective action against acute pancreatitis could be the action of berberine to counteract the oxidative damage caused by radicals or may attenuates the inflammatory infiltration induced by DBTC. Hence the berberine could be a potential source of traditional drug for acute pancreatitis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Insights into the pharmacology of GPER/GPR30 and its involvement in gallstone formation
Authors: Arnatt Christopher , Sondergard Chelsea , Wang Helen , Wang David 
Published: June 2021

The prevalence of gallstone disease is a major public health issue globally, and due to complex etiology of gallstones, there are a limited number of pharmaceuticals available. While the G protein‐coupled estrogen receptor (GPR30/GPER) has been shown to induce gallstone formation, little is known about its chemical biology and pharmacology. Building upon previous computational modeling of GPER, a novel series of antagonists were designed and synthesized. These novel compounds were assessed for their therapeutic value via calcium mobilization, cyclic AMP, and estrogen receptor a and β binding assays. From this series of compounds, one compound, SG‐1, exhibited superior antagonism and selectivity for GPER. The potential therapeutic value of SG‐1 was further validated in an in vivo mouse model of gallstone formation. Data revealed that SG‐1 treatment results in a dose‐dependent reduction in gallstone prevalence in estrogen‐treated ovariectomized mice.Support or Funding InformationSupported by startup funds from Saint Louis University (C.A.) and this work was supported in part by research grants DK73917 and DK101793 (to D.W.).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Piezo1 Channels Contribute to Flow‐Induced Intracellular Calcium Transients in Mouse Cortical Collecting Duct (CCD)
Authors: CARRISOZA‐GAYTAN ROLANDO , DALGHI MARIANELA G , APODACA GERARD L , KLEYMAN THOMAS R , SATLIN LISA M 
Published: June 2021

Within the CCD, an acute increase in tubular fluid flow rate (TFFR) (i) subjects principal (PC) and intercalated (IC) cells therein to biomechanical forces including hydrostatic pressure/circumferential stretch (CS), fluid shear stress (FSS) and drag/torque on PC cilia, and (ii) leads to a biphasic increase in intracellular Ca2+ concentration ([Ca2+]i) characterized by an immediate high amplitude increase to a peak value within ~10 s due to release of IP3‐sensitive internal Ca2+ stores coupled to extracellular Ca2+ entry at the basolateral membrane, followed by a decay to a plateau elevation in [Ca2+]i that exceeds baseline, sustained by luminal Ca2+ entry (Liu et al. AJP Renal Physiology 2003, 2005; 2007). The latter increase in [Ca2+]i is necessary for flow induced K+ secretion (FIKS) in the microperfused mammalian CCD. The identity of the basolateral Ca2+ entry path is unknown. We sought to examine whether Piezo channels, a family of mechanically‐activated Ca2+ permeable channels, contribute to this response. RNAscope detected single Piezo1 RNA molecules in mouse renal cortex. Immunofluorescence studies in mice expressing Piezo1‐tdTomato construct revealed robust basolateral and faint apical Piezo1 localization in both PC and IC. Flow‐induced [Ca2+]i transients, inferred from fluorescence intensity ratios (FIRs), were measured in microperfused mouse CCDs loaded with the Ca2+ indicator fura‐2 and then subject to low or high TFFRs, in the absence or presence of GsMTx4 (piezo‐blocker) or Yoda1 (piezo1‐activator). IC in CCDs from male and female mice preincubated with luminal or bath GsMTx4 (5 μM) failed to exhibit a flow‐induced increase in [Ca2+]i in response to an acute increase in TFFR. Addition of luminal or bath Yoda 1 (1 μM) to CCDs perfused at a low TFFR led to an increase in [Ca2+]i over 90 s in both PC and IC; addition of Yoda1 (10 μM) to the bath further increased [Ca2+]i. We speculate that mechanosensitive Piezo1 channels, expressed on the basolateral>apical membranes of PC and IC in the mouse CCD, contribute to the increase in [Ca2+]i triggered by an increase in TFFR.Support or Funding InformationNIDDK: R01 DK38470, R01 DK51391 and P30 DK079307This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial Topoisomerase Dysfunction Contributes to Myocardial Mitochondrial Dysfunction and Mitochondrial DNA (mtDNA) Degradation
Authors: Edwards John , Musial Nicole , Bradley Colleen , Sira Elizabeth 
Published: June 2021

Diabetes promotes mtDNA degradation as an antecedent event to cardiomyocyte cell death and we have reported that mitochondrial topoisomerase dysfunction participates in this pathway. In post mitotic cells, the mitochondrial genome continues to replicate about once a month and resolving the topology of mtDNA replication would be almost insurmountable in the absence of the mitochondrial topoisomerases. It remains unclear how chronic hyperglycemia induces this shift in mitochondrial topoisomerase function from physiologic to pathologic. To that end, we hypothesize that low levels of ROS alter mitochondrial topoisomerase function to accelerate mtDNA damage. DNA relaxation of supercoiled DNA is a hallmark measurement of topoisomerase function. Supercoiled DNA incubated with isolated myocardial mitochondria or purified topoisomerase 1 generated the same isopane pattern of relaxation. In the absence of mitochondrial extracts, H2O2 (125–1000 μM) did not alter DNA relaxation of supercoiled DNA. In contrast, in the presence of myocardial mitochondrial extracts, H2O2 (125–1000 μM) significantly increased DNA relaxation in a dose dependent manner. Similar results were obtained following the DNA cleavage protocol. Using the iron chelator EDTA, we determined that DNA relaxation was not due to a Fenton effect. Following immunoprecipitation using mtTop1 or Topo‐2beta antibodies, mitochondrial extracts retained DNA relaxation function indicating the overlapping presence of functional mitochondrial topoisomerases. Using isolated intact mitochondria, incubation with H2O2 significantly increased mitochondrial superoxide levels; an effect that could be blocked with MitoTempo (mitochondrial specific antioxidant). NOX4 is a member of the NADPH oxidase family, but is unique in its preferential generation of H2O2 over superoxide (O2¾). Others have reported that NOX4 is localized to the outer mitochondrial membrane. Overexpression of NOX4 in H9c2 cells exacerbated intracellular and mitochondrial superoxide levels when incubated in the present of high glucose. Diabetes increases pentose shunt pathway activity, which in turn may fuel mitochondrial localized NOX4 to promote the generation of H2O2 a diffusible ROS species. Collectively our results suggest that increased mitochondrial ROS presentation alters mitochondrial topoisomerase function. Separate from a direct impact of oxidative stress, our studies find that alteration of mitochondrial topoisomerase function accelerates and propagates an increase in mtDNA damage. These findings indicate that the functional state of the mitochondrial topoisomerases participates in the pathophysiology leading to the development of diabetic cardiomyopathy.Support or Funding InformationSupported in part by NIH RO3HD065555101 and TCUS Biomedical Education FundThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Blood Pressure Regulation Using Electroacupuncture in Middle‐Aged Hypertensive Women Associated With Mitochondrial Beta‐Oxidation
Authors: Tjen‐A‐Looi Stephanie C , Fu Liang‐Wu , Xie Lifang , Li Peng , Malik Shaista 
Published: June 2021

Acupuncture at specific acupoints reduces elevated blood pressure. We have shown that 8‐week electroacupuncture (EA) at P5‐6 and St36‐37 decreases blood pressure in men (38–75 year) with mild to moderate hypertension (HTN) more effectively (70% responsiveness, ≥ −6 mmHg). In contrast, the majority (64%) of middle‐aged hypertensive women were not responsive to this treatment. We decided to add acupoints that target the endocrine system as well (LI4, CV3‐4, SP 6, LV3, K3). However, the underlying mechanisms of how acupuncture lowers blood pressure remain unclear. Metabolomics provides a powerful method to assess the biochemical signature involved in cellular processes that are involved in human health and disease. By using untargeted metabolomics, therefore, we aimed to identify metabolic biomarkers and assess pathways involved in sympathoinhibitory EA and endocrine‐sympathoinhibitory EA treatment lowering blood pressure in middle‐aged perimenopausal females with mild to moderate HTN. Twenty four enrolled women ages 45 to 70 year with hypertension were randomly assigned to sympathoinhibitory EA treatment (P5‐6+St36‐37, n=10) or endocrine‐sympathoinhibitory EA treatment (P6‐LI4+St36‐Sp6+CV3‐4+Li3‐K3, n=14). Participants were not on antihypertensive medications and hormonal therapy. The blood pressure lowering responsiveness (≥ −6 mmHg) to endocrine‐sympathoinhibitory EA was 80% while only 33% of the subjects treated with sympathoinhibitory EA lowered their blood pressures. Peak systolic blood pressure (SBP) during 24 hours dropped from 169±4.1 to 157±3.3 mmHg (P=0.002) while averaged 24‐hour SBP dropped from 138±2.3 to 131±2.3 mmHg (P=0.001) in 14 subjects treated with endocrine‐sympathoinhibitory EA. The averaged 24‐hour mean blood pressure (MBP) and peak MBP decreased respectively with P values of 0.002 and 0.025 in the 14 subjects. On the other hand, 8‐week course of sympathoinhibitory EA treatment did not decrease SBP, MBP, peak SBP, and peak MAP in 10 other subjects, consistent with our prior study. Concurrently, metabolomics analysis indicated that the plasma acylcarnitines (short (P=0.03) and medium chain fatty acids (P=.008)) were reduced significantly in 14 subjects following 8‐week endocrine‐sympathoinhibitory EA treatment in contrast to sympathoinhibitory EA. Previous studies have shown that arterial stiffness and hypertension are associated with higher acylcarnitine levels which indicate higher beta‐oxidation associated with mitochondrial dysfunction. Endocrine‐sympathoinhibition EA enhances blood pressure lowering responsiveness in middle‐aged hypertensive women likely through decreasing acylcarnitine production, reflecting lower beta‐oxidation at the mitochondrial level.Support or Funding InformationAdolph Coors FoundationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic Cannabis Use and Circulating Bio‐Markers of Neural Health, Stress, and Inflammation in Physically Active Individuals
Authors: Lisano Jonathon , Kisiolek Jacob , Phillips Kristina , Smoak Peter , Stewart Laura 
Published: June 2021

PurposeThe purpose of this cross‐sectional study is to compare measures of general health, circulating concentrations of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), c‐reactive protein (CRP), interleukin‐6 (IL‐6) and cortisol in physically active cannabis users (CU) and physically active non‐cannabis users (NU).MethodsParticipants (N=30; n=20 male) were defined as CU (n=15; n=10 male) if they were using cannabis products at least once a week for the past 6‐months or NU (n=15; n=10 male) if they had not used any cannabis products in the past 6‐months. Age, height, weight, and body composition were assessed. Resting heart rate (HR) was obtained using a Polar heart rate monitor and VO2max, as a measure of cardiovascular fitness, was assessed with a ParvoMedics metabolic cart using a graded treadmill protocol. Fasted and rested intravenous blood samples were collected and ELISAs were used to obtain NGF, BDNF, CRP, IL‐6, and cortisol concentrations. Data are presented as mean ± SD and analyzed with a student's t‐test (alpha=0.05).ResultsCU used cannabis products an average of 4.5±2.5 times per week. There were no significant differences between CU and NU age (23.4±4.4 yrs), height (177.6±8.2 cm), weight (75.7±15.5 kg), body composition, or VO2max (50.3±7.4 ml/kg/min). HR was significantly higher in CU (72.7±15.9 bpm) compared to NU (62.5±7.9 bpm) (p=0.04). Concentrations of BDNF were significantly lower in CU (5.6±0.8 ng/mL) compared to NU (6.3±0.8 ng/mL) (p=0.02). There were no differences in concentration of NGF (193.7±71.6 pg/mL), CRP (CU=1.0± 1.83; NU=0.5±0.4 mg/L), IL‐6 (1.28±0.6 pg/mL), or cortisol (19.8±7.0 ng/mL).ConclusionsAlthough aspects of general health appeared to be similar between NU and CU groups, this study revealed elevated resting HR and reduced BDNF concentrations in CU compared to NU. Although there were no differences between groups with respect to bio‐markers of stress (IL‐6, CRP and cortisol); CU were classified as moderate risk for cardiovascular disease (CVD) compared to low CVD risk for NU as defined by CRP concentrations. This study suggests that chronic cannabis use is associated with altered central and peripheral synaptic plasticity, as well as the potential for increased CVD risk classification in physically active individuals.Support or Funding InformationThis research was funded by the University of Northern Colorado New Projects Program Grant, University of Northern Colorado Graduate Student Association and Natural and Health Sciences Student Grants.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.