bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of nanoparticle size and surface charge on uptake into rat alveolar epithelial cells
Authors: Sipos Arnold , Peti‐Peterdi Janos , Chow Robert H , Borok Zea , Kim Kwang‐Jin , Crandall Edward D 
Published: November 2023

Physicochemical properties of nanoparticles, including size, composition and surface charge, are likely to affect nanoparticle interactions with biological cells/tissues. We reported previously that uptake of polystyrene nanoparticles (PNP) from apical fluid into primary cultured rat alveolar epithelial cell monolayers (RAECM) takes place in a manner dependent on exposure time and dose. Classical endocytosis pathways do not appear to play a role in the uptake of PNP, consistent with a ‘diffusional’ process. We also reported that autophagy is involved in cellular processing of PNP, in that accumulation of PNP in lysosomes occurs via autophagic flux. Lysosomal exocytosis of PNP (modulated by intracellular [Ca2+]), takes place in parallel with a slower ‘diffusional’ egress process. In this study, we investigated the effects of nanoparticle size and surface charge on uptake into RAECM grown on Anopore filters. RAECM were exposed apically to 80 μg/mL carboxylated or amidinated 20, 100 or 200 nm PNP (with near‐infrared fluorescent label) and intracellular [PNP] was monitored over 24 hr using live cell imaging. Results showed that intracellular [PNP] reached a plateau of ~0.6 mg/mL after ≥12 hr of apical exposure to both 20 nm carboxylated or amidinated [PNP] at 80 μg/mL. As PNP size increased from 20 to 100 to 200 nm, steady state intracellular [PNP] decreased from ~0.6 to ~0.3 to ~0.1 μg/mL, respectively, after ≥12 hr. These data indicate that, for RAECM, PNP uptake is strongly dependent on PNP size, while the role of surface charge appears to be less important. Additional factors (e.g., nanoparticle‐associated protein corona) might alter the charge effect. In general, understanding the role of physicochemical properties of nanoparticles in cellular interactions will lead to improved nanoparticle design for protection from cellular injury and approaches to therapeutic utilization in drug/gene delivery.Support or Funding InformationFunding: NIH; Hastings and Whittier Foundations.
GPR171 in the Basolateral Amygdala Regulates Stress and Reward‐Related Behaviors
Authors: Bobeck Erin , Pena Darlene , Gomes Ivone , Fujita Wakako , Devi Lakshmi 
Published: November 2023

Recently we deorphanized GPR171, a hypothalamic G protein‐coupled receptor, as the receptor for the abundant neuropeptide, Big‐LEN. By virtual screening of a library of small molecules, we identified compounds that selectively target GPR171. Using these compounds we found that the activation of GPR171 within the hypothalamus regulates feeding. Additional characterization of the distribution of GPR171 revealed high expression in the basolateral amygdala. This led us to explore a role for GPR171 in various stress and reward‐related behaviors using lentiviral shRNA‐mediated knockdown of GPR171. While decreases in expression of GPR171 in the basolateral amygdala had a minor or no effect on anxiety and depression, the decrease in expression had a major effect on fear learning and morphine conditioned place preference. In addition, we investigated these behaviors using GPR171 selective ligands and found a reduction in anxiety. Interestingly, we also find that mice subjected to repeated stress show an increase in GPR171 mRNA. Together, these results suggest a role for GPR171 in stress and reward‐related behaviors within the amygdala.Support or Funding InformationThis work was supported in part by NIH grants NS026880 and DA019521 to L.A.D and a NIDA postdoctoral training grant DA007135 to E.N.B.
Adenosine A3 Receptor Expression and Function in Mitochondria
Authors: Doyle Timothy M. , Phillps Grady , Tosh Dilip K. , Gratton Michael Anne , Jacobson Kenneth , Bennett Gary J. , Salvemini Daniela 
Published: November 2023

Mitochondrial dysfunction is emerging as a common mechanism of a variety of neurodegenerative diseases, including chemotherapy‐induced neuropathic pain. We have recently found that administration of selective agonists to the G protein‐coupled, adenosine A3 receptor (A3AR) prevents the development of chemotherapy‐induced mitochondrial dysfunction in peripheral sensory neurons that result peripheral neuropathy and chronic pain. Recent evidence also indicates the presence of G proteins and G‐coupled protein receptors within mitochondria, including the adenosine A1 and A2A receptors. Bioinformatic analysis revealed potential mitochondrial‐associated sequences in published A3AR protein sequences, thus prompting us to hypothesize that A3AR is associated with mitochondria and its signaling could regulate mitochondrial function. Using Western blot analysis and transmission immune‐electron microscopy, we report for the first time that A3AR is present in normal rat mitochondria purified from liver, spinal cord and peripheral blood lymphocytes. Moreover, we also report that when normal rodent mitochondria are pretreated with the selective A3AR agonist, MRS5890, they produced more ATP following ADP‐stimulation than mitochondria treated with its vehicle (0.002% DMSO). We also find that MRS5980 counteracts normal mitochondrial membrane depolarization during calcium mobilization. Collectively, our data provide the first evidence for the presence A3AR in mitochondria and initial mechanistic insight to its functional role in mitochondria that potentially could serve as a target for the treatment of neurodegenerative disease.Support or Funding InformationNIH‐NCI CA169519
A load of hypergravity inhibits AMPKα and causes liver injury in mice, which can be prevented by preconditioning loads
Authors: Lee Sang gil , Lee Chan gyu , Kim Sang geon 
Published: November 2023

An understanding of the effects of hypergravity on energy homeostasis is necessary in managing proper physiological countermeasures to assure safe and effective aerospace missions and human survival. This study investigated whether a single or multiple load(s) of mice to hypergravity has an effect on the molecules associated with energy metabolism. In the liver, AMPKα level and its signaling pathway were repressed 6 h after a load of +9 Gz hypergravity for 1 h, and then gradually returned toward normal up to 24 h. AMPK β1 and γ1/2/3 levels were unchanged. AMPKα level was restored after 3 consecutive daily loads to +9 Gz, suggestive of adaptation of animals to repetitive preconditioning. In cDNA microarray analyses, 221 genes were differentially expressed by +9 Gz (P<0.05 and a 1.5‐fold cutoff), and the down‐regulated genes included Nrf2 targets. Nrf2 knockout abrogated the recovery of AMPKα elicited by 3 loads to +9 Gz, indicative of the role of Nrf2 in the adaptive increase of AMPKα. In addition, +9 Gz stress inhibited other transcription factors including STAT3, FOXO1/3 and CREB, and this effect was attenuated during the resting time. Similarly, apoptotic markers were enhanced with liver injury 6 h after a load of hypergravity, indicating that the liver may be vulnerable to hypergravity stress. Hepatocyte death was also abolished after multiple hypergravity loads. Our results show that a load of mice to +9 Gz hypergravity causes AMPKα repression with liver injury, which can be overcome by multiple loads to hypergravity as mediated by Nrf2.Support or Funding InformationAs supported by the NRF grant funded by the Korea government (NRF‐2014M1A3A3A02034698)
Metformin Protects against 7, 12‐Dimethylbenz[a]anthracene‐Induced Breast Cancer Initiation and Adduct Formation In Vivo Rat Model Through CYP1A1/CYP1B1 Pathway
Authors: Belali Osamah M , Attafi Ibraheem M , Moraished Bader A , Ansari Mushtaq A , Korashy Hesham M 
Published: November 2023

Recent studies have established that metformin (MET), an oral anti‐diabetic drug, possesses antioxidant activity and is effective against different types of cancer in several carcinogen‐induced animal models and cell lines. In that, a study from our laboratory showed that MET protected against 7, 12‐dimethylbenz[a]anthracene (DMBA)‐induced breast carcinogenesis in vitro human non‐cancerous epithelial breast cells (MCF10A). However, whether MET can protect against breast cancer initiation in vivo rat model has not been reported before. Therefore, the overall objectives of the present study are to elucidate the potential chemopreventive effect of MET against DMBA‐induced rat breast carcinogenesis and explore the underlying mechanism involved, specifically the role of cytochrome P4501A1/1B1 pathway. In this regard, treatment of rat with DMBA (25 mg/kg) induced initiation of breast carcinogenesis in rat as evidenced by changes in biochemical and histopathological analysis. Importantly, the protective effect of MET against DMBA‐induced breast carcinogenesis was supported by the capability of MET (500 mg/kg) to restore the induction of the mRNA and protein expression levels of several DNA repair genes ( OGG1, XRCC1), and the level of 8‐hydroxy‐2‐deoxyguanosine (8‐OHdG), a DNA damage marker. In addition, MET was able to restore DMBA‐mediated changes in the gene expression of apoptotic (caspase‐3 and BAX) and inflammatory (TNF‐α and IL‐6) mediators. Interestingly, these changes were associated with proportional decrease in DMBA‐induced CYP1A1 and CYP1B1 gene expression. In conclusion, the present work provides the first evidence that MET protects against DMBA‐iduced rat breast carcinogenesis by inhibiting DNA adduct formation, inducing apoptosis and inhibiting the expression of CYP1A1/CYP1B1 pathway.Support or Funding InformationThis Project was funded by the National Plan for Science, Technology and Innovation (MAARIFAH), King Abdulaziz City for Science and Technology, Kingdom of Saudi Arabia, Award Number (12‐MED3131‐02)
Investigate Cigarette Smoke Induced IL‐8 Secretion in Ovarian Cancer Cells and Estrogen Mediated Modulations
Authors: Sumanasekera Wasana , Beckmann Taralein 
Published: November 2023

Cigarette smoking is a global health crisis and exposure to cigarette smoke (CS) leads to variety of diseases including many cancers. Although there are few clinical reports involving CS exposure with the ovarian cancer, laboratory based reports are rare. The focus of our laboratory is to investigate whether CS contributes to the development of ovarian cancer and unveil the role of estrogen in CS induced effects. Our preliminary data shows that CS induced functional changes in ovarian cancer cells (OCCs). The objective of current work is to investigate IL‐8 release as one of the contributing mechanism for CS induced effects. The specific aims are 1). Detection of CS induced IL‐8 release 2). Investigate whether estrogen cause IL‐8 release 3) Examine estrogen's role in modulating CS induced IL‐8 release.MethodsOCCs were cultured using standard procedure. Following 10 nM estradiol pre‐treatment, OCCs were exposed to two different concentrations of CS, which is created in‐house using a published method. Following CS exposure, released IL‐8 was detected using IL‐8 Enzyme Linked Immunosorbant Assay (ELISA). ELISA was conducted based on manufacturer's instructions (Boster Bio).ResultsCS increased IL‐8 release in OCCs. 10 nM estradiol pre‐treatment also increased IL‐8 release. However, when OCCs are exposed to both estrogen and CS, estrogen pre‐treatment attenuated CS induced IL‐8 release showing a protective effectSupport or Funding InformationSullivan University Faculty Development Grant
Pancreas Proteome of Palmatine and Purified Fraction E Extract of Coscininum Fenestratum ‐Supplementation in STZ‐Induced Diabetic Rats
Authors: Okechukwu Patrick Nwabueze , Ekeuku Sophia Ogechi , Sena Teo Swee , Akuwoah Gabriel Akyirem , Froemming Gabriele Anisah 
Published: November 2023

Previous studies have reported the antidiabetic and antioxidant activity of Purified Fraction E extract from stem of Coscinium Fenestratum and Palmatine. Therefore in this study the mechanism of action of Palmatine, Fraction E and Tolbutamide were evaluated using proteomics. The proteome of pancreas harvested from STZ‐ induced diabetic rat and supplemented with Palmatine, Fraction E and tolbutamide for 90 days were evaluated using 2‐D gel electrophoresis and MALDI‐TOF MS/MS. The identified proteins which were expressed based on their functions were: chaperone/heat shock proteins (calreticulin, protein disulfide isomerase a2/a3, glucose regulatory protein 78 and heat shock protein 70); digestion protein (pancreatic amylase 2a3/1a, chymotrypsinogen, anionic trypsin 2/trypsin v‐a); lipid digesting proteins (bile salt activated lipase protein, Enoyl CoA hydratase and pancreatic lipase); channel proteins (ATP synthase B and voltage dependent anion channel2 protein); protection proteins (selenium‐binding protein1 and Lactoyl glutathione lyase) and antioxidant proteins (glutathione‐s‐transferase, peroxidoxin 4 and serum albumin). The differential protein expression of tolbutamide, Palmatine and Fraction E‐treated groups suggest that their antidiabetic effect maybe by the stimulation of chaperone/heat shock proteins, digestion protein, lipid digesting proteins, channel proteins and protection proteins. However, Palmatine and Fraction E stimulated antioxidant proteins expression, which were absent in Tolbutamide treated group.Support or Funding InformationCERVIE UCSI University
Hypoglycemic and Hypolipidemic Effects of Acalypha Wilkesiana Leaves in Alloxan Induced Diabetic Rats
Authors: Olukunle Johnny Olufemi , Jacobs Eniope Bamidele , Oyewusi Joseph Adeoye 
Published: November 2023

Hypoglycemic and hypolipidemic activities of 21day, once a day dosing of aqueous extract of Acalypha wilkesiana administered orally at a dose of 400mg/kg was studied in alloxan‐induced diabetic rats. The hypoglycemic effect of a single dose of 400mg/kg methanolic extract of Acalypha wilkesiana was also evaluated within 24 hours of dosing the rats, Glibenclamide, Polyoxyethylene sorbitan monooleate (Tween 80) and distilled water were orally administered to equal number of rats as positive and negative controls, respectively. Effect of oral administration of aqueous extract Acalypha wilkesiana (400 mg/kg) on level of blood glucose, triglycerides (TG), low density lipoprotein (LDL), high density lipoprotein (HDL) were determined.The 400 mg/kg aqueous extracts caused 50.8% reductions in blood glucose when compared to 70.6% reduction caused by Glibenclamide, a standard antidiabetic agent. The aqueous extract also caused statistically significant (p<0.05) decrease in the values of serum cholesterol (121±0.5) mg/dl, LDL (61.70±1.4) mg/dl and statistically significant (p<0.05) increase in the value of HDL (42.95±1.0) mg/dl when compared to the untreated control. The hourly percentage glucose reductions obtained, revealed a steady decrease from 10.7% at 30mins post administration to 43.8% reduction at 24 hours, this is comparable with the 52.4% reduction obtained at 24 post administration of standard antidiabetic agent by Glibenclamide. This result establishes the hypoglycemic and hypolipidemic activities of Acalypha wilkesiana leaves.
Depletion of Dietary microRNAs from Cow's Milk Causes an Increase of Purine Metabolites in Human Body Fluids and Mouse Livers
Authors: Lozano Ana Gabriela Aguilar , Baier Scott , Adamec Jiri , Sadri Mahrou , Giraud David , Zempleni Janos 
Published: November 2023

BackgroundMicroRNAs (miRNAs) play essential roles in gene regulation. We have recently reported the ground‐breaking discovery that miRNAs are not exclusively derived from endogenous synthesis but can also be obtained from dietary sources such as cow's milk. Evidence suggests that milk consumption is inversely correlated with serum uric acid (UA) concentrations, and 19 miRNAs regulate 11 pathways in purine metabolism.HypothesisConsumption of a diet depleted of milk miRNAs causes changes in purine metabolism.MethodsIn a first screen, C57BL/6 mice were fed a diet depleted of miRNAs (encapsulated in exosomes) from age 3 to 7 weeks [denoted Exosome (E) Minus, E−], whereas controls were fed a miRNA‐sufficient diet (E+). Mouse liver metabolites were analyzed using non‐targeted LC/MS‐MS, and targeted enzymatic assays for purine metabolites. Subsequently, we assessed purine metabolites in urine and plasma from human dairy avoiders and dairy consumers. The statistical significance of differences was assessed by t‐test.ResultsPrincipal Components Analysis (PCA), a dendrogram performed with hierarchical clustering, Variable Important in Projection (VIP) scores, and a heat map analysis consistently suggested that concentrations of purine metabolites were higher in E− livers than in E+ livers, and that purine metabolites were affected by milk miRNA intake to a greater extent than any of the other metabolites that were identified (p<0.01; n=5). As for effect size, the concentrations of xanthine were 16.6 ± 3.3 μM and 10.1 ± 1.0 μM in E − and E+ livers, respectively (p<0.05). Likewise, the concentrations of plasma xanthine and urinary uric acid were 81.6% and 19.3% higher in human dairy avoiders compared with dairy consumers (n=6).ConclusionDietary depletion of milk miRNAs causes an increase in purine metabolites in mouse tissues and human body fluids.Future plansA reversed‐phased HPLC was developed to quantify eight purine metabolites in a comprehensive analysis of purine metabolism; gene expression analysis and purine feeding studies will be used to identify the step(s) in purine metabolism targeted by milk miRNAs.Support or Funding InformationNIFA/USDA 2015‐67017‐23181, NIH 1P20GM104320, Gerber Foundation, Egg Nutrition Center, USDA W‐3002, Hatch and NIH P30GM103335
A Genome‐Wide Association Study of Plasma α2‐Macroglobulin Concentrations in Young Adults
Authors: Jamnik Joseph , Borchers Christoph , El‐Sohemy Ahmed 
Published: November 2023

BackgroundGluten intake has been shown to be associated with circulating levels of α2‐macroglobulin in young adults without celiac disease. Gluten‐free foods have increased in popularity over the past decade despite a poor understanding of the physiological effects of gluten intake in those without celiac disease. Plasma α2‐macroglobulin represents a physiologically important molecule involved in inflammation and cytokine release, but the contribution of common genetic variation to the observed variation in circulating concentrations of α2‐macroglobulin remains unclear.ObjectiveThe objective was to identify genetic variants that are associated with α2‐macroglobulin in a population of young adults using a genome‐wide approach.MethodsA genome‐wide association study (GWAS) was conducted on circulating levels of α2‐macroglobulin in a population of young Caucasians without clinically diagnosed celiac disease from the Toronto Nutrigenomics and Health Study (n=488). Concentrations of α2‐macroglobulin were measured in plasma using a multiple reaction monitoring HPLC‐MS/MS assay. Genome‐wide genotyping was conducted for 822,000 single nucleotide polymorphisms (SNPs) using the Affymetrix 6.0 chip. The association between genome‐wide genetic variants and plasma α2‐macroglobulin was explored using linear regression with an additive mode of inheritance.ResultsCirculating α2‐macroglobulin was associated with variation in ICOSGL (p = 3.9×10−7), a T‐cell stimulatory gene involved in cytokine release and various immune‐related disorders including celiac disease.ConclusionThese results suggest that the physiological effects of gluten intake may be mediated by genes involved in T‐cell proliferation and cytokine release.Support or Funding InformationResearch support from the Advanced Foods and Materials Network.
Interaction between PhIP from meat intake and polymorphism in superoxide dismutase 2 gene increases risk of oxidative stress
Authors: Carvalho Aline , Steluti Josiane , Carioca Antonio Augusto , Loureiro Ana Paula , Gattas Gilka , Fisberg Regina , Marchioni Dirce 
Published: November 2023

BackgroundHeterocyclic amines (HCA), especially 2‐Amino‐1‐methyl‐6‐phenylimiazo[4,5‐b]pyridine (PhIP) from meat intake have been linked to cancer due to oxidative stress generated during the metabolism. This process is mediated by detoxifying enzymes, as superoxide dismutase 2 (SOD2), which is encoded by a polymorphic gene. The presence of this polymorphism Val16Ala (rs4880) decreases the enzyme activity increasing oxidative stress and risk for chronic disease as cancer. The aim of the present study was to investigate if there is an interaction between PhIP from meat intake and SOD2 polymorphism on biomarkers of oxidative stress.MethodsWe performed a cross‐sectional study in adults and older adults from Brazil (n=239). PhIP intake was assessed using one 24‐hour dietary recall and a structured questionnaire with cooking methods and levels of meat doneness. It was used PCR‐based assays to detection SOD2 polymorphism. Malondialdehyde was measured in plasma after derivatization by thiobarbituric acid and separation on HPLC. Multiple linear regression models were performed to verify the association and interaction between PhIP intake and SOD2 on risk of oxidative stress. Sensitivity analyses were performed in a subsample (n=79) using a DNA adduct marker, 8oxodG measured by ELISA, as a dependent variable.ResultsThe frequency of the minor allele (C) was 45%. PhIP mean intake was 254ng/day (95% CI: 200;308ng/day). MDA concentration was associated with PhIP intake (β=0.04 P=0.004) after adjustment for energy and fruit intake, sex, age, BMI, race, physical activity, C reactive protein, global methylation, and smoking. We also noted a significant interaction between SOD2 polymorphism and PhIP intake on MDA (P=0.035). Subjects who had one or two risk alleles presented a significant relationship between PhIP and MDA concentration (β=0.06 P=0.01) compared with who had the wild‐type genotype (β=0.01 P=0.870). Moreover, we confirmed a significant interaction between SOD2 and PhIP intake on 8oxodG (P=0.022) after adjustment.ConclusionPhIP was associated with oxidative stress and interacted with polymorphism in SOD2 gene increasing oxidative stress and DNA adducts in a general population.Support or Funding InformationResearches relating to this abstract were funded by Municipal Health Secretariat of Sao Paulo, National Counsel of Technological and Scientific Development and Sao Paulo Research Foundation