bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Sulfiredoxin‐Peroxiredoxin axis promotes urethane‐induced lung adenocarcinoma through the regulation of the tumor microenvironment
Authors: Hao Yanning 
Published: May 2022

Lung adenocarcinoma is the most common type of lung cancer and has a strong association with tobacco smoking. Smoking leads to the accumulation of reactive oxygen species (ROS) and oxidative stress damages to macromolecules, leading to abnormal cell growth & proliferation, transformation, changes in cell metabolism and a variety of other physiological functions. Accumulation of ROS only leads to increased expression of cellular antioxidants that contribute to tumorigenesis and cancer progression. Sulfiredoxin (Srx) is the only enzyme that reduces over‐oxidized forms of typical 2‐Cys Peroxiredoxins (Prxs). Compared to other Prx family members, Srx preferentially interacts with and has a higher binding affinity to Prx4. Our previously work demonstrated that the Srx‐Prx4 axis enhances the RAS/RAF‐MEK‐ERK signaling cascade to promote lung cancer cell proliferation and metastasis. However, the role of the Srx‐Prx axis in de novo lung tumorigenesis has not been explored.In this study, Prx4‐/‐ mice and Srx‐/‐Prx4‐/‐ mice were generated and mouse lung carcinogenesis was induced by a well‐established urethane protocol. Briefly, wildtype and knockout mice at 8‐week of age were injected intraperitoneally with 1g/kg of urethane once per week for three successive weeks. Ten weeks after the administration of urethane, all mice were euthanized and examined for tumors. Our results indicate that both knockout mice have significantly reduced rates on tumor incidence, multiplicity, and size. Compared with tumors from wildtype mice, the rate of cell proliferation in tumors from either knockout mice is significantly decreased.Macrophages are the most abundant immune cells identified in the tumor microenvironment of solid tumors and their presence correlates with reduced survival in most cancers. In addition to all other markers, we also compared the presence of two distinct states of polarized macrophages including the classically activated macrophage (M1) and the alternatively activated macrophage subsets (M2) in urethane‐induced tumors using specific surface marker staining. We found that there are significantly increased numbers of M1 cells in tumors of either Prx4‐/‐ or Srx‐/‐Prx4‐/‐ mice compared with those of wildtype mice. In contrast, there are significantly few numbers of M2 cells in tumors from either knockout mice.It has been shown that M1 macrophages have immune‐stimulatory properties through the expression of a series of pro‐inflammatory cytokines, chemokines, and effector molecules, whereas M2 cells express a wide array of anti‐inflammatory molecules. Therefore, the difference in tumor‐associated macrophages caused by the loss of Prx4 or Srx and Prx4 may contribute to the reduced lung tumorigenesis observed in the mouse model. In addition, bioinformatic analysis shows that Prx4 is overexpressed in the majority of cancers, and the increased expression of Srx and Prx4 both show a strong correlation with poor prognosis in lung adenocarcinoma patients.
Effect of BAY‐117082, a NLRP3 inflammasome inhibitor, in a mouse model of nitroglycerin (NTG)‐induced migraine
Authors: Cuzzocrea Salvatore , Campolo Michela , Scuderi Sarah A. , Filippone Alessia , Lanza Marika , Casili Giovanna , Paterniti Irene , Esposito Emanuela 
Published: May 2022

Migraine is a common brain disorder characterized by recurrent episodes of headache. It is a complex and multifactorial disease with a higher prevalence in females than in males. Several risk factors have been associated to migraine disease as genetic factors, sex and age. The pathophysiology of migraine is still unclear; however, it has been proven that NOD‐like receptor protein 3 (NLRP3) inflammasome pathway overactivation as well as p‐ERK/p‐CREB axis contribute to migraine pathogenesis. Therefore, the aim of this study was to investigate the effect of BAY‐117082, a NLRP3 inflammasome inhibitor and p‐ERK/p‐CREB modulator, in an in vivo model of nitroglycerin (NTG)‐induced migraine.Migraine model was induced by NTG intraperitoneal administration at dose of 10 mg/kg diluted in 0.9% saline. Mice were treated intraperitoneally with BAY‐117082 at doses of 1 mg/kg, 5 mg/kg, and 10 mg/kg, 5 minutes after NTG injection. Mice were sacrificed 4 h following NTG injection; the whole brain with the rostral spinal cord was removed to perform several analysis.Our results demonstrated that, following behavioral tests for pain and photophobia, BAY‐117082 treatment at doses of 5 mg/kg and 10 mg/kg reduced pain attacks induced by NTG more than BAY‐117082 at the dose of 1 mg/kg. Moreover, the treatment with BAY‐117082 at doses of 5 mg/kg and 10 mg/kg significantly reduced histological damage in the trigeminal nerve nucleus and significantly decreased inflammosome activation by reduction of NLRP3, ASC, IL‐1β and TNF‐α expression. Additionally, the treatment with BAY‐117082 at doses of 5 mg/kg and 10 mg/kg significantly modulated p‐ERK/p‐CREB axis activation through the reduction of p‐ERK, p‐AKT, p‐CREB and p‐PI3K expression.Therefore, the obtained results offer new insight into the role of NLRP3 inflammasome pathway and p‐ERK/p‐CREB axis in migraine pathogenesis, suggesting that BAY‐117082 could be considered a novel therapeutic strategy to treat migraine.
Global PER1 knockout Dahl Salt Sensitive rats show increased expression of renal Edn1 mRNA and Endothelin‐1 peptide
Authors: Juffre Alexandria , Costello Hannah , Crislip G. Ryan , Douma Lauren A. , Cheng Kit‐Yan “. , Bratanatawira Phillip , Zietara Adrian , Dissanayake Lashodya V. , Staruschenko Alexander , Gumz Michelle L. 
Published: May 2022

BackgroundPreviously, we showed that global knockout of the circadian clock protein PERIOD1 (PER1) in Dahl Salt Sensitive (SS) rats resulted in worsening of the SS hypertensive phenotype (1). PER1 KO rats also exhibited evidence of renal injury and increased plasma aldosterone levels after three weeks on a high salt diet (4% NaCl). Because we previously showed that PER1 negatively regulates the peptide hormone Endothelin‐1 (ET‐1) via transcriptional control of the ET‐1‐encoding gene Edn1, we wanted to investigate ET‐1 regulation in this novel rat model. Interestingly, Edn1 is also positively regulated by aldosterone through the mineralocorticoid receptor (MR).HypothesisGiven that ET‐1 signaling can be pro‐hypertensive and profibrotic in the kidney, we hypothesized that ET‐1 peptide levels in the kidney may be upregulated as part of the phenotype of the PER1 KO SS rats.MethodsProtein and total RNA were isolated from the kidney samples of control and PER1 KO SS rats on normal diet (0.4% NaCl) or after three weeks on a high salt (4% NaCl) diet. Renal ET‐1 peptide and plasma aldosterone levels were measured by ELISA and quantitative PCR was performed to quantify the renal mRNA levels of ET‐1 and its receptors ETA and ETB, MR, and GR. Renal MR protein levels were measured by Western Blot and quantified by relative densitometry. Data were evaluated for genotype or diet differences using ANOVA.ResultsRenal ET‐1 peptide levels were significantly higher in PER1 KO rats on high salt diet when compared to control SS rats (p=0.0003), as were plasma aldosterone levels (p=0.0156). There was also a significant genotype difference in Edn1 mRNA levels, with increased levels in PER1 KO rats on a high salt diet (p=0.0389). In contrast, significant differences were not observed in the mRNA levels of ET‐1 receptors Ednra and Ednrb between genotypes or diets. Importantly, quantification of renal MR protein levels showed marked increase with genotype and treatment effects (genotype: p=0.0002, treatment: p=0.0133).ConclusionsThese data suggest that PER1 KO results in increased ET‐1 in the kidney, due to increased transcription of the Edn1 gene, with increased plasma aldosterone and renal MR protein levels. This apparent negative regulation of ET‐1 by PER1, coupled with increased MR and aldosterone levels, is likely to contribute to the hypertensive and renal injury phenotype of PER1 KO SS rats on a high salt diet. Together these data show an important role for PER1 and its target ET‐1 in salt‐sensitive hypertension and the accompanying renal injury.Reference1 Spires DR, Zietara A, Levchenko V, Gumz ML, Staruschenko A. Knockout of Per1 Exacerbates the Hypertensive Phenotype of the Dahl Salt Sensitive Rat. FASEB J. 2020;34(S1):1.
Metabolic Profile in Neonatal Pig Hearts
Authors: Li Pengsheng , Tang Ling , Zhu Wuqiang 
Published: May 2022

BackgroundCardiomyocytes in postnatal hearts undergo hypertrophy to adapt to increased blood pressure and volume. The process of cardiomyocyte maturation is associated with a cessation from cell cycle and a series of metabolic switches. Prior research has shown metabolic switches in developing mouse hearts. However, metabolic changes in postnatal pig hearts have not been reported.MethodsWe evaluated the metabolic profile in pig hearts at postnatal day 1 (P1), day 3 (P3), day 7 (P7), and day 28 (P28) (n=6 each, both males and females; Yorkshire‐Landrace Cross background) using a targeted liquid chromatography tandem mass spectrometry assay. Expression of lactate dehydrogenase isoforms was evaluated by western blotting.ResultsThere is a clear separation of the detected metabolites in P1 versus P28 hearts. Active anabolisms of nucleotide and proteins were observed in P1 hearts when cardiomyocytes retain high cell cycle activity. Abundance of the intermediate metabolites in the pathways of glycolysis was decreased in P28 comparing to P1 hearts, which implicates an overall reduced level of glycolysis. The decreased abundance of intermediates in TCA cycle and increased abundance of lactic acid in P28 hearts suggested that glycolysis rather than glucose oxidation is the main form of glucose metabolism in mature pig hearts. Reduced abundance of GLCN6P was observed in P28 hearts compared to P1 hearts. Increased abundance of hydroxyproline was observed in P1, P3, and P7 hearts compared to P28 heart.ConclusionPostnatal cardiomyocyte maturation is associated with metabolic switch from glucose to fatty acids, active posttranslational protein modification and ECM remodeling.
Increased Insulin Resistance in Male Hepatic Androgen Receptor Knockout Mice Fed High Fructose Diet
Authors: Carr Kiana R. , Wilson Andre , Levy Josephine , Falzarano Claire , Lofton Taylor , Andrisse Stanley 
Published: May 2022

IntroductionInsulin resistance is a common hepatic dysfunction amongst individuals suffering from diabetes or obesity and affects 10‐25% of the general population1. The intracellular signaling pathway is reliant on every step being executed correctly. It is evident that excess dietary fats and sugars will result in obesity and likely diabetes. This cellular machinery can be altered when there is a build‐up of lipids which impact molecular insulin signaling. Our objective is to determine if chronic intake of fructose by male liver androgen receptor knockout (Liv‐ARKO) mice will show cascade alteration resulting in increased resistance to insulin when compared to male Liv‐ARKO mice on a control diet. We hypothesized that LivARKO mice on HFrD would show impaired insulin action when compared to control and chow diet mice.MethodsWe placed male Liv‐ARKO mice on high fructose (HFrD), Control (Research Diets Inc), and Chow diets. We performed insulin tolerance tests (ITT), glucose tolerance tests (GTT), and pyruvate tolerance tests (PTT) after 1‐2 months of being on diets. The mice were later sacrificed, and their tissues were extracted for further tests. To assess differences in insulin action that may arise from increased fructose some mice were given a dose of 0.5 U/kg insulin before being sacrificed to further investigate insulin signaling proteins. BCA protein assays were done to ensure that protein concentration in each sample was standardized. Western blots were performed using tissue from liver to measure molecular insulin action, via p‐AKT presence.ResultsMale mice on control diet for 2‐months showed improved glucose tolerance and enhanced pyruvate tolerance when compared to male mice on control diet for 1 month. Mice on 2‐month control diet showed impaired insulin sensitivity compared to mice on the same diet for 1 month.Male LivARKO mice displayed no insulin stimulated p‐AKT when on a chow diet. Levels of p‐AKT were the same in basal and insulin male LivARKO mice fed a chow diet. Male LivARKO mice fed a control diet displayed a 4‐fold increase in levels of p‐AKT when stimulated by insulin compared to mice not given insulin (basal). Moreover, male LivARKO mice fed a HFrD displayed no insulin stimulated p‐AKT. Foxo1 levels were not altered by insulin stimulation in male LivARKO mice fed a chow, control, or HFrD. However, it should be noted that control and HFrD both increased Foxo1 levels compared to chow diet.ConclusionOur results suggest that hepatic insulin action or the ability for insulin to initiate glucose uptake in liver is decreased in male LivARKO mice on HFrD. Further investigation of Research Diets Inc. data is necessary to determine why the control diet and HFrD lead to the results discovered.
Sleep Apnea and Kidney Disease: The Role of Hypoxia and Lysyl Oxidase
Authors: Thomas Ann , Jakkidi Nishka R. , Li Shiyu , Cheng Kit‐Yan , Gumz Michelle , Canales Muna T. , Mohandas Rajesh 
Published: May 2022

Study Objective/HypothesisChronic kidney disease (CKD) or progressive loss of kidney function, is a serious health condition that affects 1 in 7 adults or an estimated 37 million Americans. Epidemiological studies have linked CKD with Sleep Apnea (SA), a breathing disorder characterized by erratic breathing, and episodes of hypoxia. However, the mechanisms that underlie the association between CKD and SA are not well understood. Hypoxia Inducible Factor‐1α (HIF‐1α) is a transcription factor that regulates the body’s response to hypoxia. Lysyl oxidase and lysyl oxidase likes (LOX and LOX‐Ls) are a group of copper enzymes that are hypoxia responsive and pro‐fibrotic. We hypothesized that hypoxia in SA upregulates HIF‐1α to drive expression of LOX or LOX‐L enzymes, fibrosis, and CKD.Materials & MethodsSamples from the Sleep and Nephrology and Outcomes Research Study (SNORE), a prospective observational study of sleep disorders in veterans with CKD, were used for this study. The patients were divided into three main groups: CKD with no SA (n=14), CKD with severe SA (n=12) and healthy controls (n=13). RNA from peripheral blood mononuclear cells (PBMC) was isolated using PAXGene Blood RNA tubes (Qiagen). The RNA was then used as a template for reverse transcription, converting it into cDNA and fold changes in gene expression between groups were examined using real time PCR. Differences between groups were compared using nonparametric Mann Whitney U tests and a p‐value < 0.05 was considered statistically significant.ResultsLOX expression was lower in patients with CKD compared to healthy controls, regardless of SA (CKD with SA: 0.28 ± 0.06 (p=0.04) and CKD without SA: 0.33 ± 0.07 (p=0.09)). There were no significant differences seen in any of the LOX‐L expression (LOX‐L1 (p=0.84), LOX‐L2 (p=0.35), or LOX‐L3 (p=0.93)). Similarly, HIF‐1α was lower in patients with CKD (CKD with SA: 0.69 ± 1.0 (p=0.47) and CKD without SA: 0.46 ± 0.06 (p=0.01)). There was no statistically significant correlation between LOX and HIF‐1α (r=0.04, p=0.83).ConclusionOur results showed that HIF‐1α and LOX expression in PBMC was decreased in patients with CKD regardless of if they had SA or not. The mechanisms underlying these observations, the functional significance, and if these changes are representative of tissue LOX expression needs further studies.
Agreement Between MRI, Ultrasound, and Histology in Detecting Size Changes of the Vastus Lateralis Following Resistance Training
Authors: Ruple Bradley A. , Smith Morgan A. , Osburn Shelby C. , Sexton Casey L. , Godwin Josh S. , Stock Matt S. , Poole Chris N. , Roberts Michael D. , Young Kaelin C. 
Published: May 2022

HypothesisWe hypothesize that MRI‐determined vastus lateralis (VL) cross‐sectional area (CSA) will increase following resistance training and that changes in VL size determined by ultrasonography and histology methods will agree with and/or be related to MRI‐determined changes in VL size.MethodsTwelve healthy, untrained, college‐aged males (20±1 y; BMI: 26.9±5.4 kg/m2) had VL assessments via MRI, B‐mode ultrasound (thickness and cross‐sectional area) prior to (PRE) and after 10 weeks of resistance training (POST). Muscle biopsies were obtained at the site of imaging at PRE and POST for mean fiber cross‐sectional area (fCSA) determination. Agreement between MRI mCSA (mCSAMRI) and panoramic‐based ultrasound mCSA (mCSAUS) at PRE and POST as well as change scores in these variables were examined using Bland‐Altman plots for mean bias and 95% limits of agreement (LOA) as well as Lin’s concordance correlation coefficients (CCC). The relationship between fCSA with mCSAMRI, mCSAUS, and B‐mode ultrasound thickness (VLThick), as well as VLThick and mCSAMRI were tested with Pearson’s correlations.ResultsBoth mCSAMRI and VLThick significantly increased following training (28.1±6.1 to 31.1±6.9cm2 (p=0.005) and 2.35±0.35 to 2.63±0.39cm (p=0.001), respectively). mCSAUS and fCSA did not significantly increase with training (26.3±6.4 to 28.3±7.5cm2 (p=0.064), and 4896±901.3 5443.2±1449.3µm2 (p=0.12), respectively). Bland‐Altman analysis revealed a small mean bias in mCSAUS relative to MRI (‐2.29 m2, 95% LOA: ‐8.74, 4.15 cm2). Concordance between mCSAUS and mCSAMRI taken at PRE and POST time points were excellent (CCC=0.83). Bland‐Altman analysis revealed good agreement between change scores in mCSAUS and mCSAMRI (‐1.08cm2, 95% LOA: ‐5.49, 3.33 cm2). There was a moderate concordance between mCSAUS and mCSAMRI change scores (CCC=0.70). A positive, moderate correlation was found between percent change in both VLThick and mCSAMRI as well as VLThick and mCSAUS (r=0.39, p=0.071 and r=0.52, p=0.080, respectively). A negative correlation was found between percent changes in fCSA and mCSAMRI (r= ‐0.569, p=0.054). Finally, correlations were not significant between percent changes in VLThick and fCSA (r=0.053, p=0.871) or fCSA and mCSAUS (r= ‐0.271, p=0.394).ConclusionThese data suggest that, relative to MRI, panoramic ultrasound is a reliable method to track mCSA changes of the VL. However, fCSA changes do not associate with tissue level changes, and this discordance warrants further research.
Exposure to Eosinophilic Esophagitis Limits Esophageal Carcinogenesis in a Murine Model
Authors: Fuller Anne , Kabir Mohammad F. , Karami Adam , Klochkova Alena , Mu Anbin , Klein‐Szanto Andres , Hamilton Kathryn , Whelan Kelly 
Published: May 2022

IntroductionEpidemiological data suggest that eosinophilic esophagitis (EoE) patients do not develop esophageal cancer. Here, we paired murine models of EoE and esophageal squamous cell carcinoma (ESCC) to perform the first functional interrogation of the relationship between EoE and esophageal cancer.MethodsC57B6 mice were treated with MC903/Ovalbumin (OVA) to induce EoE and the esophageal carcinogen 4‐nitroquinoline 1‐oxide (4NQO) to induce ESCC according to the experimental scheme shown in Fig 1A. Esophagi of mice treated with vehicle controls (n=6), MC903/OVA alone (n=4), 4NQO alone (n=10), and MC903 in combination with 4NQO (n=5) were dissected. Tumor frequency was recorded and tumor load (tumor size x tumor number) was determined under a dissecting microscope. H&E stained tissues were evaluated for esophageal lesions. CD45‐positive immune cells were isolated from the spleens of mice with EoE or naïve controls then co‐cultured with the murine‐derived esophageal cell line AKR. After 24‐72 hours of co‐culture, live/dead dye was used to assess AKR viability.Results10/10 mice treated with 4NQO alone developed ESCC as expected in this robust tumor model. In mice co‐treated with MC903/OVA and 4NQO, ESCC was detected in 4/5 mice with a significant decrease in tumor load as compared to mice treated with 4NQO alone (Fig 1B, C). 6/6 mice treated with MC903/OVA alone and 4/4 mice treated with propylene glycol (vehicle for 4NQO) failed to display tumors (Fig 1C). We further evaluated esophageal epithelium of 4NQO‐treated animals to characterize distribution of ESCC lesions. While squamous dysplasia (pre‐malignant), early ESCC and late ESCC were present in all tumor‐bearing animals; there was a decrease in early ESCC and a lack of late ESCC lesions in 4NQO‐treated mice that were exposed to MC903/OVA‐induced EoE (Fig 1D). Moreover, the total percentage of esophageal epithelium occupied by neoplastic lesions was significantly diminished in mice co‐treated with 4NQO and MC903/OVA (Fig 1E). To investigate how EoE limits esophageal carcinogenesis, we exposed AKR esophageal tumor cells to CD45‐positive cells isolated from spleens of strain‐matched donor mice with or without EoE (Fig 2A). We found a robust induction of cell death in tumor cells exposed to immune cells from EoE donor mice compared to those exposed to immune cells from vehicle‐treated controls at 48 hours (n=3, p<0.05) (Fig 2B, C).ConclusionsThese data indicate that exposure to EoE activates an anti‐tumoral immune response that limits esophageal carcinogenesis. These studies have the potential for significant implications related to the clinical care of patients affected by EoE and ESCC.
Pulmonary Hypertension Induces Sex Differences in Control of Contracting Rat Skeletal Muscle Oxygen Pressures
Authors: Schulze Kiana M. , Weber Ramona E. , Horn Andrew G. , Colburn Trenton D. , Ade Carl J. , Poole David C. , Musch Timothy I. 
Published: May 2022

Pulmonary hypertension (PH) is a disease characterized by pulmonary vascular remodeling, cardiac impairments, and skeletal muscle dysfunction. Males and females display different features of the disease, including a higher incidence in females, while males present with greater mortality. Exercise intolerance is a cardinal manifestation of PH, but any potential differences regarding skeletal muscle function during contractions between males and females have not been studied. This investigation tested the hypothesis that sexual dimorphism exists in the oxygen delivery (Q̇O2)‐to‐oxygen utilization (V̇O2) matching with PH as represented by oxygen partial pressures (PO2) at rest and following the onset of contractions. Considering that females display greater reliance upon nitric oxide (NO) for vascular function and PH decreases NO bioavailability, we hypothesized that PH would attenuate the skeletal muscle PO2 response to a greater extent in females than males.MethodsThirty‐three Sprague‐Dawley rats were randomly assigned to healthy (HC) or PH groups. PH rats (male: n=7; female: n=9) were administered monocrotaline (MCT; 50 mg/kg, i.p.) while HC rats (male: n=9; female: n=8) were given saline. Disease progression was monitored via echocardiography. Phosphorescence quenching determined the O2 partial pressure in the interstitial space (PO2is) in the spinotrapezius muscle at rest and during contractions.ResultsAll PH rats displayed criteria for disease including right ventricular (RV) hypertrophy, elevated RV systolic pressure, and pulmonary congestion compared to HC. Indicators of PH severity did not differ between males and females (P>0.05). Females displayed a greater reduction in baseline PO2is in PH versus HC when compared to male counterparts (‐29 ± 3% vs. ‐18 ± 2%; P<0.05). No sex differences were present in PO2is kinetics (i.e., tau, amplitude) throughout contractions. However, females with PH reached a lower nadir in PO2is than PH males (4.2 ± 0.3 vs. 7.4 ± 0.4 mmHg; P<0.05).ConclusionsThese findings support that Q̇O2‐to‐V̇O2 matching is impaired in a sex specific manner in skeletal muscle. Individual contributions of O2 delivery and utilization, as well as biochemical analysis of NO in skeletal muscle will be invaluable to elucidating the mechanistic bases for the effects seen herein.