bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reversal of myofibroblast differentiation by phorbol 12‐myristate 13‐acetate is PKC‐independent
Authors: Talbott George , Le Helen , Lor Paia , Jin Zhuqiu 
Published: June 2021

Fibrosis often results in organ dysfunction and failure in diseases, such as: chronic heart failure, hepatic cirrhosis, pulmonary fibrosis, and end‐stage renal disease. The differentiation of fibroblasts into myofibroblasts results in the secretion of collagens and other extracellular matrix proteins that limit organ function and underlie the fundamental basis of fibrosis. TGF‐β1 induces the differentiation of α‐SMA‐expressing myofibroblasts from fibroblasts. However, the mechanism to induce reversal of myofibroblast differentiation remains elusive. Phorbol 12‐myristate 13‐acetate (PMA) has been used for the stimulation of lymphocytes and splenocytes. It is also involved in multiple cellular functions by potently activating protein kinase C (PKC). Nevertheless, the effect of PMA on the formation of myofibroblasts is unknown. To investigate whether PMA plays a role in myofibroblast differentiation, NIH 3T3 fibroblasts were cultured in DMEM medium for 48 hours in the presence of DMSO (vehicle control), PMA, or PKC inhibitors (calphostin C, chelerythrine, and staurosporine) alone or plus PMA. The expression of α‐smooth muscle actin (SMA, a hallmark of the myofibroblastic phenotype), fibroblast‐specific protein 1 (FSP1, a biomarker of fibroblasts), and TGF‐β1 in cultured fibroblasts was detected by western blotting and immunofluorescence. Expression of α‐MARCK phosphorylation was also detected to examine the activity of PKC activation. Positive staining of α‐SMA was observed in some cells in control culture conditions, and these α‐SMA positive cells exhibited significant morphology changes with large nuclei and cytoplasm, compared with α‐SMA negative fibroblasts. Treatment with PMA 50 ng/mL for 48 hours reduced the expression of α‐SMA and TGF‐β1, and pretreatment with the three PKC inhibitors mentioned above could not abrogate PMA‐induced reduction of α‐SMA and TGF‐β1. These results indicate that some myofibroblasts are derived from fibrolbasts under basic culture conditions. PMA induces reversal of myofibroblast differentiation via a PKC‐independent mechanism.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects Of Nitrite In Goldfish (Carassius Auratus) Olfactory Function
Authors: Martinez Melody , Huertas Mar 
Published: June 2021

In the last decade, freshwater ecosystems have experienced a dramatic increase of nitrogenous compounds as a result of farming and industrial activity. Aquatic ammonium (NH4+) is naturally transformed to nitrite (NO−2) by nitrifying bacteria. Nitrite is toxic to vertebrates and is known to cause disruption of physiological processes such as ion homeostasis, respiration, and cardiovascular activity. However, little is known about the effects of NO−2 on the olfactory function in teleost, which is critical for the survival of the species. This study aims to determine the physiological effects of sub‐lethal concentrations of nitrite in the olfactory function of goldfish (Carassius auratus). Thus, goldfish were exposed to 6 different NO−2 concentrations (1000 mM, 500 mM, 300 mM, 100 mM, 10 mM and 0 mM.) for 72 h. At the end of the experiment, blood, brain and nose samples were obtained for histological analysis. Additionally, in order to determine the reduction of olfactory function, olfactory responses to several food odorants were recorded by electro‐olfactogram (EOG) after exposure of the nose to several NO−2 concentrations. No mortality was observed in any treatment. Goldfish exposed to the two highest concentrations of NO−2 exhibited metahemoglobemia (darkening in the blood due to irreversible oxidation of hemoglobin). Histological analysis of the olfactory epithelium showed an increase in mucus production and mucus cells. Moreover, the olfactory epithelia of these fish showed hyperplasia of the olfactory sensory neurons. Hyperplasia of epithelium cells occurred in goldfish exposed to high NO−2 concentrations. Preliminary electrophysiology recordings showed a decrease in the olfactory function due to NO−2 treatments. These results suggest that sub‐lethal concentrations of nitrite are neurotoxic and alter the olfactory function in fish.Support or Funding InformationResearch support: Texas State University start‐up funding to Dr. Mar Huertas.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Clinical Implication of Compounds which could induce the function of human organic cation transporter 2 (hOCT2), while inhibit the function of human Multidrug and Toxin Extrusion Proteins 1 (hMATE1)
Authors: YANG HONG , Guo Dong , Shu Yan 
Published: June 2021

The human organic cation transporter 2 and human multidrug and toxin extrusion protein 1 (hOCT2/hMATE1) transport system is a pair of transporter proteins highly expressed at the basolateral and apical membrane of renal proximal tubules respectively. They function collaboratively to facilitate elimination of a variety of compounds into urine, thus playing a crucial role in drug disposition and response. Preliminary data from our lab showed that acute exposure (20 minutes) of Cd2+ could significantly upregulate the activities of human hOCT2, while inhibit the function of hMATE1, both in a dose‐dependent manner (Fig. 1A&1B). Beside Cd2+, we identified imatinib, an anticancer drug, could induce the function of hOCT2 while inhibit the function of hMATE1 (Fig. 2). In vivo study also showed that pre‐treatment of imatinib could significantly increase the accumulation of metformin in mice kidney while had no impact on the plasma exposure of metformin (Fig. 3A&3B). It implicated that the index of plasma drug exposure will not be able to correctly reflect drug‐drug interaction when a drug could induce hOCT2 while inhibit hMATE1 at the same time, which potentially could cause significant drug accumulation in kidney tubular cells.Support or Funding InformationNIH: R01GM099742FDA: U01FD004320This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction between Adenosine and β‐adrenergic receptors in regulating vascular tone in mouse mesenteric arteries.
Authors: Yadav Vishal R. , Zhang HanTing , Mustafa S. Jamal S. 
Published: June 2021

Adenosine (AR) and β‐adrenergic receptors (βR) are GPCRs involved in the regulation of cardiovascular function. Adenosine is an endogenous nucleoside which controls local blood flow through activation of four subtypes of ARs. Stimulation of A1 and A3 ARs produce contraction while A2A and A2B ARs cause relaxation in mouse aorta and mesenteric arteries (MAs). Stimulation of β1R causes increase in heart rate and contractile force whereas β2R produces relaxation of blood vessels.Reportedly, A1AR, β1R, and β2R form heterodimers in human heart. Hetero‐dimerization between A1AR and βR leads to altered receptor pharmacology, functional coupling, and intracellular signaling pathways. Importantly, stimulation of the myocytes with isoproterenol (βR agonist) inhibits the effect of A1AR activation on the co‐localization of protein kinase C‐ɛ involving protein kinase A. Thus, a complex interaction between signaling cascades of A1AR and β1R exists, which may regulate cardiovascular function. In the present study, we aimed to determine whether interaction between β1R and ARs exists in regulating vascular tone in mouse aorta and MAs.We measured mean arterial blood pressure (MABP), protein expression, organ bath (aorta) and DMT (MA) muscle tension in β1R KO and WT mice. Our non‐invasive CODA tail cuff BP (mmHg) data showed significant reduction in MABP in β1R KO mice. The MABP in β1R WT mice was 83.4 ± 3; whereas 68.9 ± 3 (p<0.05) in β1R KO mice. Our DMT experiments revealed that β1R was partly involved in the vascular relaxation response to isoproterenol. Significantly reduced relaxation response to isoproterenol was observed in A1AR KO mice MAs. Importantly, significantly enhanced contraction to CCPA (A1AR agonist) was observed in β1R KO mice MAs. These contractile responses were blocked by HET0016 (Cyp4A inhibitor). NECA (non selective ARs agonist) induced responses were comparable in MAs. Furthermore, our vascular studies revealed significantly reduced PE and ACh induced contraction in MAs of β1R KO mice whereas KCl induced responses were comparable. Our western blot analysis revealed higher A1AR and comparable Cyp4A expression levels in MAs. Our studies in aorta revealed comparable CCPA induced contraction in β1R WT and KO mice. In addition, comparable A1AR and Cyp4A expression levels and vascular responses to KCl, ACh and PE were observed in aorta.In conclusion, our data revealed a possible interaction between β1R and A1AR at a vascular level in MAs. This unique receptor cross‐talk may enlighten the important signaling responses controlling vascular function.Support or Funding InformationSupported by HL 027339This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rapamycin PFC Nanoparticles Pre‐Treatment Mitigates Cisplatin Induced Acute Kidney Injury
Authors: Doherty Justin , Fazal Jawad , Wickline Samuel , Pan Hua 
Published: June 2021

Cisplatin, belonging to the class of platinum‐containing anti‐cancer drugs, is widely used in a variety of cancers, including sarcomas, lymphomas, small cell lung cancer, squamous cell carcinoma of the head and neck, bladder cancer, testicular cancer, cervical cancer and ovarian cancer. Despite its chemotherapeutic efficacy, cisplatin induces acute kidney injury (AKI) in approximately one‐third of treated patients, which further impairs their quality of life, and thwarts successful outcomes of cisplatin treatment. Although it is known that early enhancement of autophagy and inhibition of cisplatin induced inflammation is beneficial in mitigating cisplatin induced AKI, treatment targeting both mechanisms remains to be developed. Accordingly, our hypothesis is that rapamycin PFC nanoparticles could serve this purpose via rapamycin targeting of mTOR signaling to induce autophagy and inhibit its downstream NF‐κB signaling pathway, a convergent gate keeper in inflammation. Rapamycin delivery by PFC nanoparticles enables sufficient drug delivery to the site of injury with minimum systemic exposure as well as sustained local drug release.Fluorescence microscopic images demonstrated the delivery of rapamycin PFC nanoparticles to the kidneys, where they remained for 24 hours following i.v. injection (n=3). Therefore, we performed rapamycin PFC nanoparticles (1 ml/kg) treatment 24 hours before cisplatin injection (10 mg/kg) (n=5). Mice receiving cisplatin injection only (n=5) and mice receiving no injection (n=5) served as controls. On the day of rapamycin PFC nanoparticles and cisplatin injections as well as 24, 48, and 72 hours after the cisplatin injection, blood was collected from each mouse for a BUN test to assess kidney function. The results (Fig. A) illustrated that cisplatin treatment induced mild kidney function impairment, which was mitigated by rapamycin PFC nanoparticles pre‐treatment. Furthermore, using proximal tubule cells (NRK‐52E), we demonstrated that the benefit of rapamycin PFC nanoparticles pre‐treatment could come from 1) enhancing autophagy (Fig. B), where p62 level was reduced in the groups with the rapamycin nanoparticles treatment regardless of cisplatin treatment; 2) preventing cisplatin induced inflammation (Fig. C) through NF‐κB signaling pathway activation, where p65 upregulation was only observed in the group receiving cisplatin treatment alone; and 3) inhibiting apoptosis (Fig. C), where rapamycin pre‐treatment inhibited cisplatin induced Bax expression.In conclusion, rapamycin PFC nanoparticles pretreatment could potentially mitigate cisplatin induced AKI in cancer patients.Support or Funding InformationThis work was supported by the NIH grant R01 DK102691.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Brain Microvascular Endothelial Derived Microparticles as a Non‐Invasive Diagnostic Tool for Multiple Sclerosis
Authors: Yun J. Winny , Boyer Chris , Minagar Alireza , Alexander J. Steven 
Published: June 2021

BackgroundMultiple sclerosis (MS), an autoimmune‐neurodegenerative disease, exhibits features of neurovascular inflammation including endothelial activation and shedding of microparticles (MPs). While the brain lacks ‘classical’ lymphatics, we previously reported expression of venous‐lymphatic endothelial markers (LYVE‐1, podoplanin, prox‐1, VEGFR‐3, VEGF‐C, VEGF‐D) in both brain tissue and sera, with altered expression of these proteins in MS. Based on differential expression of these markers in brain tissue, we propose that these markers may originate within brain microvascular endothelium and undergo changes in disease which can be analyzed diagnostically.ObjectivesTo validate the neurolymphatic biomarker matrix as a novel, non‐invasive and cost‐effective tool for diagnosing MS and discriminating MS subtypes.MethodsNeurolymphatic biomarker expression was determined in human cerebral microvascular endothelial cells (hCMECs) following 24–48 hours of TNF‐α (20 ng/ml) exposure +/− IFN‐γ (1000 U/ml) using western blotting. MPs from these cells were visualized using scanning electron microscopy. Human sera were analyzed by flow cytometry, western and dot blotting to characterize relative partitioning of neurolymphatic biomarkers in MP.ResultshCMECs released MPs that contain lymphatic and junctional markers as well as caveolin‐1. Plasma MPs were seen to liberate soluble (non‐particulate) neurolymphatic biomarkers upon storage, indicating that serum represents a more stable MP‐derived pool of these markers. MP flow analysis of intracellular actin revealed that permeabilization destabilizes phosphatidylserine‐Annexin V interaction which is used to designate MP identity, suggesting that flow cytometry of MPs may require more judicious interpretation. All neurolymphatic markers so far tested were able to distinguish MS from healthy control serum samples; FOXC2 expression can further distinguish subtypes of MS: RRMS from SPMS.ConclusionLymphatic biomarkers in plasma and sera are expressed by brain microvascular endothelial cells and exhibit changes in expression in response to inflammatory stimuli (cytokines). Such alterations parallel transfer of these markers into caveolin‐1 ‐enriched microparticles, which represent a circulating ‘snapshot’ of the vascular surface that can be analyzed by several immune approaches, particularly following storage‐dependent solubilization of these biomarkers.Support or Funding InformationSupported by the Annette Funicello Research Fund for Neurological Diseases and the Malcolm Feist Predoctoral Fellowship from Center for Cardiovascular Diseases and Sciences, LSU Health Sciences Center‐ShreveportThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Carotid Arterial Stiffness Predicts White Matter Lesion Volume in Older Adults
Authors: Balestrini Christopher S. , Al‐Khazraji Baraa K. , Lingum Navena R. , Vording Jennifer L. , Suskin Neville , Shoemaker J. Kevin 
Published: June 2021

Markers of arterial wall stiffness are used commonly as indicators of cardiovascular risk. Stiff conduit vasculature transmits pulse waves with increased velocities, which may lead to end‐organ microvascular damage. However, the association of peripheral vascular stiffness to the brain's vulnerability for vascular impairment has not been established. In the brain, this microvascular damage could include breakdown of the blood‐brain barrier causing leakage of fluid through the cerebrovascular endothelial layer. Fluid‐attenuated inversion recovery (FLAIR) imaging allows the visualization of fluid‐abnormalities in subcortical tissue, which can be quantified as white matter lesions (WML). These WML have been linked to cognitive impairment, and labelled as risk factors for stroke, dementia, and cardiovascular mortality. This research tested the hypothesis that peripheral arterial stiffness predicts levels of WML volume. We examined normotensive controls (CTRL; n = 12; 6 males; age: 42–69 years) as well as normotensive ischemic heart disease patients (IHD; n = 17; 14 males; age: 40–75 years). Common carotid artery diameters were measured during systole and diastole (B‐mode ultrasound images with ECG‐gating), and time‐aligned corresponding blood pressure values were used for measures of arterial strain (strain), arterial stiffness index (β), carotid distensibility, carotid compliance, and arterial elastic modulus. All measures were taken in the supine posture. Magnetic resonance FLAIR imaging at T2 was used to assess periventricular WML volume (WMLv) using SPM software. The IHD and CTRL cohorts had similar WMLv (4.4 ± 3.2 vs. 3.2 ± 3.5 mL; p = 0.33). Also, all participants were normotensive. Therefore, the data were pooled (n = 29) to examine the association between WMLv and peripheral arterial stiffness. Following multiple linear regressions while adjusting for age and BMI, WMLv was correlated with arterial stiffness index (r2 = 0.39, p = 0.001), strain (r2 = 0.37, p = 0.002), carotid distensibility (r2 = 0.36, p = 0.004), carotid compliance (r2 = 0.26, p = 0.009), and absolute change in carotid diameter across one cardiac cycle (r2 = 0.24, p = 0.01). Regardless of the age, BMI, and participant cohort, structural periventricular WMLv correlated with markers of arterial stiffness. These findings support the emerging hypothesis that systemic arterial stiffness affects damage in the microvasculature of the brain. Further, the resulting subcortical structural damage is apparent in older populations regardless of ageing and vascular pathology.Support or Funding InformationSupported by the Canadian Institute of Health Research (201503MOP‐342412‐MOV‐CEEA).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ZEB1 REGULATES MULTIPLE ASPECTS OF ENDOTHELIAL QUIESCENCE DURING VASCULAR REMODELLING
Authors: Bates David O. , Fernandes Cristiano S. , Usman Kiren Yacqub , Bourne Joshua , Collier Pamela , Ali Shahzeb , Kaira Spandan , Gregory Scott , Pang Vincent , Allen Claire , Grabowska Anna M. , Denning Chris , Benest Andrew V. 
Published: June 2021

Generation of new blood vessels is essential for vascular remodeling in exercise to provide greater blood flow to developing muscles. As blood vessels grow, the sprouting endothelial tips need to recognise each other, and move from a motile, angiogenic phenotype to a maturing, quiescent phenotype,, form tubes and allow blood flow to occur and regulate their development. They grow in response to cytokine signalling such as VEGF and other angiogenic factors, but they mature as a consequence of cell contact and flow. As they do so, they change their metabolic state from growing, glycolytic, to quiescent oxidative metabolism. The mechanisms behind the transcriptional control of this phenotype are unknown, but recent work has demonstrated the dynamic interplay between ‘quiescence’ factors (such as FOXO1 expression) and expression of glycolytic enzymes such as PFKFB3 through c‐MYC. We investigated potential drivers of this process and identified Zeb1 (a transcription factor with known functions in cancer progression) to be expressed by quiescent (non‐angiogenic) endothelial cells and reduced in angiogenic endothelial cells in the developing murine retina. In vitro analyses demonstrates that Zeb1 is expressed by confluent monolayers of Human Vein Umbilical Cord Endothelial cells (HUVECs) but is downregulated in actively growing cells. Zeb1 knockdown promotes an angiogenic EC phenotype: including enhanced PFKFB3, c‐myc and VEGFR2 and reduced FOXO1, Tie2 and VE‐cadherin expression. Consistently, Zeb1 knockdown EC were metabolically more active (both through oxidative and glycolytic pathways). Furthermore, loss of Zeb1 promoted EC migration and proliferation in vitro. In vivo shRNA knockdown of Zeb1 led to altered vessel formation and hypercellularity. In conclusion, Zeb1 appears to be linked to key quiescence regulating signaling pathways and furthermore regulates aspects of these pathways. Zeb1 expression appears to be a critical component for endothelial cells to generate functional vasculatures that can support enhanced muscle blood flow, by its control of FOXO1, PFKFB3 and Myc.Support or Funding InformationThis work was supported by the British Heart Foundation (grant PG/11/67/29067), the University of Nottingham Cancer Research Priority Area, and the Medical Research Council Newton Fund (MR/N01247X/1)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.