bluuebunny/crossref_metadata_2025 huggingface dataset:
Translation For Portuguese And Validation Of The Questionnaire Of The Patient Neuropathy Interpretation (PIN)
Authors: Reis MM , Parisi MCR , Giannella‐Neto D
Published: June 2021

Background

The instruments that could evaluate and extend self‐care carried on are in demanding for the prevention of chronic complication of diabetes. The Patient Interpretation of Neuropathy (PIN) has been developed and validated in the USA and in the UK and comprises a questionnaire for the qualitative measurement of comprehension of the patient with diabetes at risk to develop diabetic foot due to neuropathy and micro/macrovascular diseases. By means of this instrument, the patient can self‐evaluate her/his foot and health care provided to her/him, aiming the prevention of ulcer diabetic foot.

Objective

The aim of the present study was to translate to Portuguese validating the PIN and to assess the degree of comprehension regarding patient awareness to her/him clinical condition.

Methods

Hundred patients from the outpatient Clinic of Insensitive Foot at the Institute of Orthopedics and Traumatology, University of Sao Paulo, were surveyed by the application of PIN to determine awareness and knowledge regarding risks and consequences of and prevention of diabetic foot. Score data were expressed as mean and standard deviation and statistical analysis was performed by parametric and non‐parametric tests when needed. Statistical significance was set at p<0.05.

Result

The representative values of the assessment of the level of patients' understanding are presented in Figure 1, below. ID, identity of the disease; C, concerning; CC, cure; TL, timeline; CONS, consequences; MS, emotion. There were significant statistical differences when Sao Paulo data were compared to that obtained in UK and USA centers in most of the domains. For agreement analysis, the Spearman correlation coefficient was used. The level of significance was set at P <0.05.

Conclusion

The results of this study suggest that the use of the PIN provides better understanding of patients' condition regarding diabetic foot risk. Moreover, PIN can improve the awareness of the patients to carry on simple measures performed at home, such as, moisturize the feet, use appropriate shoes, visit the health service regularly, and daily monitoring and inspecting the feet. The application of a questionnaire as the PIN serves both to assess the patient's self‐care, but also allows the health professional delineates measures of effective prevention.

funding source: Universidade Nove de Julho ‐ UNINOVE

Support or Funding Information

UNIVERSIDADE NOVE DE JULHO ‐ UNINOVE table

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessment of Tobacco Users in Creating an Educational Intervention to Promote Oral Health
Authors: Chawla Sudeep , Einstein George P. , Tulp Orien L.
Published: June 2021

The purpose of this study was to characterize smoking behaviors amongst individuals of various socioeconomic status, to assess the need to develop an educational program that addresses the failure of getting treatments for diseases that stem from tobacco related products. Further, we wished to identify the target populations that might be the most suitable to be recruited in such a program for behavioral intervention and consultation by an oral health professional with a goal to to dissuade individuals from smoking and other tobacco use. Consuming tobacco in form of cigarettes, but also in more ‘artisanal’ forms like cigars and pipes as well as the current trend to ‘vaping’ or even consuming entirely smokeless tobacco are behaviors that are ultimately detrimental to the oral and subsequent physical health of individuals. Socioeconomic status has been negatively correlated with the engagement in such unhealthy behaviors, thereby potentially burgeoning society with increased treatment costs while suggesting that better access to public resources might be a viable way to reduce tobacco consumption across the socioeconomic strata in the future.

Methods

In a retrospective study, adult patients at NSU Dental School's Department of Oral Diagnosis were asked about their smoking behavior and several other demographic details, including age, gender, educational status and smoking history using an Axium questionnaire. Results indicated there was a slight prevalence of non‐smokers vs. smokers among the participants, with a few individuals preferring smokeless tobacco or electronic cigarettes. The prevalence or non‐smokers vs. smokers was reflected across the majority of demographic categories, with the exception of young age, old age, middle to low income and either low or the highest degree of education.

Conclusion

These results are sufficient to select a first cohort of individuals who were younger and in a lower socioeconomic category to participate in a consultation/behavioral intervention program for tobacco users with low vs. high socioeconomic status.

Support or Funding Information

Support from Institutional Resources

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Pistachios and Mixed Nuts on Bone Density and Osteogenic Gene Expression in Young Male Rats
Authors: Wickman Brooke E. , Rochester Eric , Hauffe Laura , Kern Mark , Hong Mee Young , Liu Changqi , Hooshmand Shirin
Published: June 2021

Nut consumption has beneficial health effects that include reducing the risk of cardiovascular disease, improving blood lipid profiles, and reducing oxidative damage. Nuts contain several nutrients that are involved in the maintenance and development of bone such as calcium, protein, fatty acids, and various phenolic antioxidants. To investigate the effect of nuts on bone health, thirty 21‐day‐old Sprague Dawley rats were randomly assigned to one of three isocaloric diets containing pistachios (8.1% w/w), mixed nuts (7.5% w/w), or a control diet for 8 weeks. True bone densities of tibias and femurs were measured using Archimedes' principle, and areal bone densities of total body without heads were measured using dual‐energy X‐ray absorptiometry (DXA). Femoral mRNA expression of various genes including RANK, RANKL, and OPG were also assessed. True densities of the tibias were significantly different among groups (p<0.03). Post hoc analyses indicate that tibial true densities of rats in the pistachio group were greater than tibial true densities of rats in the control group (p<0.05, 95% CIΔ 0.001 to 0.092). There was no significant difference between the tibial true densities of the rats in the pistachio group and the mixed nut group nor in the control group and the mixed nut group. There was no significant difference in the femoral true density or the whole body DXA bone mineral density among groups. Though there was a higher tibial true bone density in the pistachio group compared to that of the control group, there were no significant differences in the genes presumed responsible for these changes. It can be concluded that pistachio intake may have beneficial effects on bone density. The potential mechanism of action should be further investigated.

Support or Funding Information

American Heart Association

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of N‐Methyl‐D‐Aspartate Receptor (NMDAR) in Co‐morbid Schizophrenia and Substance Abuse
Authors: Desai Rajeev I. , Puhl Matthew D. , Coyle Joseph T.
Published: June 2021

Both schizophrenia (SZ) and substance abuse (SA) exhibit significant heritability. Moreover, N‐methyl‐D‐aspartate receptors (NMDARs) have been implicated in the pathophysiology of both SZ and SA. We hypothesize that the high prevalence of co‐morbid SA in SZ is due to dysfunction of NMDARs caused by shared risk genes. We used transgenic mice with a null mutation of the gene encoding serine racemase (SR), the enzyme that synthesizes the NMDAR co‐agonist D‐serine, to mimic an established genetic risk factor for SZ. We determined the effect of NMDAR hypofunction resulting from the absence of D‐serine on motivated behavior using intracranial self‐stimulation (ICSS) and neurotransmitter release in the nucleus accumbens using in vivo microdialysis. Compared to wild‐type mice (WT), SR−/− mice exhibited similar baseline ICSS thresholds but were less sensitive to the threshold‐lowering (rewarding) and the performance‐elevating (stimulant) effects of cocaine. While basal dopamine (DA) and glutamate release were elevated in the nucleus accumbens of SR−/− mice, cocaine‐induced increases in DA and glutamate release were blunted. γ‐Amino‐butyric acid (GABA) efflux was unaffected in the SR−/− mice. Together, these findings suggest that the impaired NMDAR function and a consequent decrease in sensitivity to cocaine effects on behavior are mediated by blunted DA and glutamate responses normally triggered by the drug. Preliminary data from ongoing behavioral studies in SR−/− and WT mice also show a decreased sensitivity to nicotine's behavioral effects. Projected to humans, NMDAR hypofunction due to mutations in SR or other genes impacting glutamatergic function in SZ may render abused substances less potent and effective, thus requiring higher doses to achieve a hedonic response, resulting in elevated drug exposure and increased dependence/addiction.

Support or Funding Information

This research was supported by NIH grant RO1MH51290‐18 to JTC, DA031231 K01 NIH/NIDA grant to RID, and a NARSAD Young Investigator Award to MDP.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Studies on identification of genes associated with neuronal memory using of rat hippocampal neurons
Authors: Opoku Ruth , Ahmed Ijaz , Patwary Mohsin , Nur‐e‐Kamal Alam
Published: June 2021

We have previously demonstrated that extracellular matrix (ECM) proteins facilitate neurite extension of rat cerebellar granule neurons (CGNs) in culture. We have also identified the functional domain(s) of these proteins involved in neurite outgrowth. Hippocampal neurons play an important role of storing memory in animals. We propose a hypothesis that “hippocampal neurons can be used to identify genes associated with storage of memory in animals”. In this report we developed a ECM‐coated culture surface to growth primary rat hippocampal neurons and determined their neurite outgrowth. We have also compared the neurite outgrowth of CGN and hippocampal neurons. Cerebellar granule neurons, cortical neurons (CN), and hippocampal neurons were plated onto the cover slips coated with extracellular matrix proteins (laminin‐1, fibronectin, and collagen) at a density of 105 neurons per ml. The neurons were allowed to extend neurites for 24 hrs in NBM/B27/KCl. The extent of neurite outgrowth was then determined using carboxyfluorescein diacetate (CFDA) labeling. Images of the cultures were captured and analyzed with the NIH Image software. A sample of 100 neurons with well‐defined processes were studied for each condition. Laminin and fibronectin‐coated surfaces induced neurite outgrowth of Cerebellar granule neurons, cortical neurons, and hippocampal neurons. It was found that collagen stimulates neuirte outgrowth of CGN, while collagen inhibited neurite outgrowth of hippocampal neuron. We have developed a condition for culture of hippocampal neurons. By immunostaining, immunoblot, and RT‐PCR analysis we found alteration of expression of some genes genes involved in neurite outgrowth, Results of these studies with be presented in this report. These hippocampal neurons will be used to apply different types of stimuli and identify genes involved in saving memory in neurons. Identification of genes associated with saving memory might be useful in developing a therapy for neurological disorders including Alzheimer's, Parkinson's diseases.

Support or Funding Information

NIH‐RISE

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Five Different Phosphorylation Sites in the C‐Terminal regulate a1b‐Adrenergic Receptor Signaling and Regulations
Authors: Hernández David Alejandro
Published: June 2021

The G protein‐coupled receptors (GPCRs) transmit a variety of signals across the cell membrane and regulate cellular activity via the intermediary role of guanine nucleotide regulatory proteins (G proteins). Among these receptors, the a1B‐Adrenergic (a1B‐AR) receptor mediates the effects of catecholamines, like epinephrine and norepinephrine, through coupling to Gq. The activation of the a1B‐AR causes polyphosphoinositide hydrolysis catalyzed by phospholipase C (PLC), generating inositol trisphosphate and diacylglycerol, which together lead to calcium signaling and protein kinase C (PKC) activation. Subsequently, receptors are phosphorylated, triggering their desensitization and internalization, a process mediated by members of the G proteins‐coupled receptor kinases (GRK) family and/or by PKC.

It is currently known the a1B‐AR is phosphorylated in two separate groups of serine residues in the carboxyl terminal tail, one by PKC (S396 and S402) and one by GRK (S406, S410 and S412) during the desensitization and internalization of the receptor. The mutation of these residues prevents agonist induced desensitization and blocks the receptor internalization into the plasma membrane.

Because the role of these specific sites remains unknown in different down‐process of the a1B‐AR, we investigated the importance of phosphorylation by PKC and GRK in calcium desensitization signal, mitogen activated protein kinases (MAPK) pathway and receptor phosphorylation. To achieve this,, we generated three site specific mutants of the a1B‐AR in which the serine residues phosphorylated by the two kinases were mutated to alanine, alone or in tandem. We found that any of these mutations in the phosphorylation sites produces changes in the heterologous desensitization of the receptor detected by intracellular calcium response and in the activation of the MAPK, when compared to the wild type. Finally, we obtained clear differences in the phosphorylation pattern of each mutant during noradrenaline stimulation.

Support or Funding Information

Partially supported by Grants from CONACYT (253156 and 882) and DGAPA (IN200915).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Different Phosphorylation Sites Regulate α1D‐Adrenergic Receptor Function
Authors: Rosas Gabriel Carmona
Published: June 2021

Human α1D‐adrenergic receptors (α1D‐AR) are seven transmembrane‐spanning proteins that mediate the actions of noradrenaline in homeostasis maintenance as well as the physiology of hypertension and benign prostatic hyperplasia. Although several studies have demonstrated that α1D‐AR is a phosphoprotein, the phosphorylation sites and the specific roles that they play in the function and regulation of this receptor, are currently unknown. Using a combination of in silico analysis, mass spectrometry and site directed mutagenesis, we identified for the first time distinct phosphorylation patterns on α1D‐AR during noradrenaline (homologous) and non‐related agents‐mediated (heterologous) desensitization. We were able to identify that G protein‐coupled receptor kinase‐2 (GRK2) and protein kinase C α/β (PKC α/β) phosphorylate α1D‐AR during these processes. Phosphorylation of the receptor's intracellular domains, such the intracellular loop 3 or the carboxyl tail, was enough to altered calcium desensitization signals, mitogen activated protein kinases (MAPK) pathway and regulate receptor internalization, suggesting that modular regulatory mechanisms heavily rely on the phosphorylation bar code and the receptor's intracellular domains.

Subsequently, we generated six specific non‐phosphorylatable α1D‐AR mutants to delve into the functions of the phosphorylation sites located only at the receptor's carboxyl tail. We found clear differences in the phosphorylation pattern of each mutant during noradrenaline and non‐related agents‐stimulation. Moreover, we analyzed the activation of the MAPK and AKT pathways as well as intracellular calcium mobilization. Modification of any of these phosphorylation sites altered the response when compared to the wild type, but no clear differences were observed among the different α1D‐AR mutants. Interestingly, seven specific phosphorylation sites clustered into two groups in the carboxyl tail, regulate the α1D‐AR subcellular location. This was confirmed when these two clusters were mutated with negative charged residues and the α1D‐AR was located again at the plasma membrane.

These data suggest that different phosphorylation sites located in the carboxyl tail regulate the α1D‐AR functions and regulation at different levels. This knowledge highly contribute to unveil new strategies to improve current pharmacological treatments and to avoid drug side effects during sustained administration.

Support or Funding Information

Partially supported by Grants from CONACYT (253156 and 882) and DGAPA (IN200915).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Amyloid Beta Peptide Modulates the Signaling Property of Chemokine‐like Receptor 1
Authors: Ye Richard D. , Zhang Shuo , Liao Dan , Gong Hao
Published: June 2021

OBJECTIVE

Amyloid beta peptides (Aβ) are the main constituents of the amyloid plaques found in the brains of Alzheimer patients. While Aβ is proinflammatory and contributes to the development of Alzheimer's disease, its signaling property remains unclear. The present study investigates how Aβ modulates signaling of one of its receptors, the chemokine‐like receptor 1 (CMKLR1, ChemR23).

METHODS

Cells expressing CMKLR1 were incubated with Aβ42 before stimulation with another agonist of the CMKLR1, the C9 peptide. Calcium mobilization, cAMP inhibition and MAP kinase activation were measured. Intramolecular FRET was determined using CMKLR1 constructs containing an ECFP attached to the C‐terminus.

RESULTS

Binding of both Aβ42 and the C9 peptide induced CMKLR1 internalization, but only the Aβ42‐induced receptor internalization involved clathrin‐coated pits. Likewise, Aβ42 but not C9 stimulated β‐arrestin2 translocation to plasma membranes. A robust Ca2+ flux was observed following C9 stimulation, whereas Aβ42 was ineffective even at micromolar concentrations. Despite its low potency in calcium mobilization assay, Aβ42 was able to alter C9‐induced Ca2+ flux with potentiation at concentrations as low as 0.1 – 1 nM. However, at higher concentrations the effect was diminished. Similar effects were seen with the C9‐induced ERK phosphorylation but the dose curve was different from that of Ca2+ flux and cAMP inhibition, suggesting a reciprocal regulatory mechanism. Intramolecular FRET assay confirmed that Aβ42 modulates CMKLR1 rather than its downstream signaling pathways.

CONCLUSION

These findings suggest that long‐term exposure to Aβ42 may alter the signaling properties of its receptors, thereby influence the pathology of Alzheimer's disease. The reciprocal regulation of CMKLR1 by Aβ provides a novel mechanism by which a ligand dose‐dependently modulates the response of its receptor to another ligand.

Support or Funding Information

Supported by grants SRG2015‐00047‐ICMS‐QRCM and MYRG2016‐00152‐ICMS‐QRCM from University of Macau and by grant 31470865 from NSFC.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of a comprehensive profiling method for regulatory lipid mediators using UPLC‐MS/MS
Authors: Yang Jun , Wan Debin , Sun Jia , Hammock Bruce D.
Published: June 2021

The regulatory lipid mediators are biologically active metabolites derived from polyunsaturated fatty acids such as the metabolites by cyclooxygenases, lipoxygenases, and cytochrome P450 enzymes. These pathways represent over 75% of pharmaceutical in mass in the world. By measuring endogenous metabolic responses to various environment stimulus, pathological processes and dietary supplements, lipidomics/metabolomics provide the most useful information about human responses. The knowledge gained from this type of research is very useful for many research fields including biomarker discovery, biotechnology advancement, discovery of novel biochemical mechanism, and assessment of clinical effectiveness.

We developed a sensitive and comprehensive profiling method for the regulatory lipid mediators covering more than 150 lipid mediators including prostaglandins, leukotrienes, epoxy fatty acids, dihydroxy fatty acids, specialized pro‐resolving mediators, endocannabinoids, nitro‐fatty acids, poly‐unsaturated fatty acids and the trace for major species of phospholipids.

Support or Funding Information

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Key Role of Soluble Epoxide Hydrolase in the Pathogenesis of Parkinson's Disease
Authors: Ren Qian , Ma Min , Yang Jun , Nonaka Risa , Yamaguchi Akihiro , Ishikawa Kei‐ichi , Kobayashi Kenta , Murayama Shigeo , Saiki Shinji , Akamatsu Wado , Hattori Nobutaka , Hammock Bruce D. , Hashimoto Kenji
Published: June 2021

Parkinson's disease (PD) is a chronic and progressive neurodegenerative disorder. Although there are many medications available to treat the symptoms in PD patients, these medications cannot reverse the progress of the disease. The soluble epoxide hydrolase (sEH) plays a key role in the inflammation which is involved in the pathogenesis of PD. The present study was undertaken to examine whether sEH plays a role in the pathogenesis of PD. MPTP (1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine)‐induced dopaminergic neurotoxicity in the striatum and substance nigra (SNr) of mice could be attenuated by subsequent repeated administration of TPPU, a potent sEH inhibitor. Furthermore, deletion of sEH gene protected against MPTP‐induced neurotoxicity in the striatum and SNr. Moreover, the treatment with TPPU or deletion of sEH gene attenuated and protected the MPTP‐induced ER stress and oxidative stress in the striatum. In addition, expression of sEH protein in the striatum from MPTP‐treated mice and patients with dementia of Lewy body (DLB) was higher than controls. Overexpression of sEH after AAV injection into the striatum significantly enhanced MPTP‐induced neurotoxicity in this region. Interestingly, expression of sEH mRNA in the neuron from iPS cells from PD patients was higher than that of healthy control subjects, and treatment with TPPU could protect against neurotoxicity in the neurons from iPS cells of PD patients. The present results suggest that sEH plays a key role in the pathogenesis of PD, and that epoxy fatty acids, their mimics as well as sEH inhibitors could be potential therapeutic or prophylactic drugs for PD.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of Structural‐Activity Features of Long Chain Polyunsaturated Fatty Acid Epoxides in Angiogenesis
Authors: Betts Rebekah Marie , Lee Kin Sing , Cinelli Maris , Karchall Lalitha , Luca Joe , Confer Nick
Published: June 2021

The cytochrome P450‐derived fatty acid epoxides are bioactive lipids with regulatory roles in a wide range of homeostatic and pathophysiological events. These lipid mediators are metabolized efficiently by the mammalian soluble epoxide hydrolase enzyme (sEH) to corresponding 1,2‐diols. Inhibition of sEH has been shown to be beneficial to human health via increasing concentrations of a variety of short‐lived epoxylipids in vivo (2). Identifying the most potent polyunsaturated fatty acid (PUFA) epoxide metabolites will provide insight on the effects of sEH inhibition. Recent studies have shown that the omega‐6 epoxyeicosatrienoic acids (EETs) from arachidonic acid are pro‐angiogenic, while the omega‐3, docosahexaenoic acid (DHA)‐derived epoxydocosapentaenoic acids (EDPs) inhibit angiogenesis, tumor growth, and metastasis (1). Given these findings, we are interested in exploring the roles of various long chain PUFA epoxides in angiogenesis. We tested a library of omega‐3 and omega‐6 epoxides in a newly developed high‐throughput tube formation assay and found their effects are regiospecific and stereospecific; Of note, the 19(S),20(R)‐EDP stereoisomer was a less effective inhibitor of endothelial cell tube formation than its respective racemic mixture. Here, we report the angiogenic activities of diverse omega‐3 and omega‐6 fatty acid epoxides that could be present in mammals from the diet, including the less explored epoxides from both omega‐3 and omega‐6 docosapentaenoic acid (DPA). We will discuss the structure‐activity features of the most potent epoxide species and also present methodology to synthesize and purify the regioisomers and stereoisomers of each series of epoxides. Ultimately, understanding activities of sEH substrates in specific regulatory environments will guide development of targeted sEH modulators that promote the beneficial effects of PUFA metabolites in human health and disease.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mesenchymal Stem Cells Ameliorate Uric Acid Induced Nephropathy in Rats
Authors: Li Lan , Cheng Dongqi , Yuan Yujia , Chen Younan , Cheng Jingqiu , Lu Yanrong
Published: June 2021

Background

The elevation of serum uric acid (UA) is positively correlated to the incidence and mortality of kidney diseases by evoked inflammation and epithelial‐mesenchymal transition (EMT) in renal tubular epithelial cells. Mesenchymal stem cells (MSCs) are able to facilitate immunomodulation and repair kidney injury in experimental models of acute or chronic kidney injury. Our study aims to investigate the effects of MSCs on kidney injury both in rats model of uric acid nephropathy and high uric acid‐treated renal tubular epithelial cells (NRK‐52E).

Methods

In vivo, rats were intragastric administrated with potassium oxonate (PO, 1500 mg/kg/day) plus adenine (Ad, 50 mg/kg/day) for 4 w to develop hyperuricemia. MSCs treatment (tail vain injection of MSCs 3×106 cells/rat) was conducted from the 2nd week for 2 times with one week interval. After 6 weeks, the kidney tissues were collected for further detections. In vitro, NRK‐52E cells were co‐cultured with or without MSCs in Transwell system. Cell cycle and apoptosis were analyzed by flow cytometry. The mRNA and protein expression of genes related to inflammation and fibrosis were detected by q‐PCR and WB.

Results

MSCs alleviated kidney injury in rats of uric acid nephropathy. Compared with UA group, HE and Masson staining have shown that the histological abnormalities and fibrosis were significantly ameliorated after MSCs treatment. Moreover, the mRNA of FN, TNF‐α, TGF‐β and IL‐1β showed marked decrease in UA + MSCs group in comparison to UA group.

MSCs inhibited cellular damage in high uric acid‐treated NRK‐52E. Comparing with high uric acid group, MSCs co‐culture induced remarkable increase of cell proliferation index (62.61% vs. 54.55%), slight decreased of apoptosis rate (3.7% vs. 4.8%) and increased expression of anti‐apoptotic protein Bcl‐2. In respect of the expression of inflammation and fibrosis related protein, MSCs co‐culture slightly downregulated the expression of TNF‐α, α‐SMA and FN, while upregulated the expression of E‐cad, but no significant change was observed on VCAM‐1.

Conclusion

During the development of hyperuricemia, MSCs effectively ameliorated inflammatory infiltration, suppressed the fibrosis progress trigged by inflammatory factors and fibrosis factor as TGF‐β, resulting in the improvement of high uric acid induced kidney lesion. MSCs exerted the protective role through increase of cell viability and proliferation, decrease of apoptosis and the expression of inflammatory factor TNF‐α, and increase the expression of E‐cad, which contributed to the suppression of interstitial transformation of renal tubular epithelial cells.

Support or Funding Information

This study was supported by the Program of National Natural Science Foundation of China (81370824), and the National Key Clinical Project.
Proximal Tubule Beta‐2 Adrenergic Receptor Is Responsible for Recovery of Renal Function Following Ischemia Reperfusion Injury
Authors: Cameron Robert Bruce , Simmons Epiphani , Miller Siennah , Gibbs Whitney , Schnellmann Rick
Published: June 2021

Acute kidney injury (AKI) is the transient loss of renal function following an insult, such as nephrotoxicant exposure, sepsis, or ischemia. Numerous cell types play a role in the pathogenesis of AKI, including immune, endothelial, and renal proximal tubule cells. Our laboratory demonstrated that the beta‐2 adrenergic receptor (ADRB2) agonist formoterol accelerates the recovery of renal and mitochondrial function in mice following ischemia‐reperfusion injury (IRI). However, the cell type(s) responsible for this recovery remains unknown. To assess the role of renal proximal tubule cells in formoterol‐induced recovery of renal function, we generated a proximal tubule‐specific knockout of the ADRB2 receptor (γGT‐Cre: ADRB2Flox/Flox). Compared to controls (ADRB2Flox/Flox), renal cortical mRNA expression of the ADRB2 receptor was decreased 90% in γGT‐Cre:ADRB2Flox/Flox mice. These mice were subjected to renal IRI, followed by once daily dosing with 0.3 mg/kg formoterol or vehicle beginning at 24 h and euthanasia at 144 h. At 24 h following IRI, ADRB2Flox/Flox and γGT‐Cre: ADRB2Flox/Flox mice had a similar loss of renal function as measured by serum creatinine (0.89 mg/dL and 0.86 mg/dL, respectively). At 144 h following IRI, both ADRB2Flox/Flox and γGT‐Cre:ADRB2Flox/Flox mice treated with vehicle exhibited a modest but incomplete recovery of renal function as measured by serum creatinine (0.43 mg/dL and 0.46 mg/dL, respectively). These mice had similarly suppressed renal cortical mRNA and protein expression of markers of mitochondrial homeostasis such as NDUFB8, COX1, ATPSβ, and Mfn2. Following treatment with formoterol, ADRB2Flox/Flox mice exhibited accelerated recovery of renal function as measured by serum creatinine (0.20 mg/dL) with associated rescue of mitochondrial markers in the renal cortex. In contrast, formoterol did not enhance the recovery of renal function as measured by serum creatinine (0.39 mg/dL) or mitochondrial markers in γGT‐Cre:ADRB2Flox/Flox mice subjected to IRI. These data reveal that formoterol‐induced ADRB2 receptor activation on proximal tubule cells induces mitochondrial biogenesis and restores renal function following AKI.

Support or Funding Information

R.B.C. is supported by F30 DK104550 and T32 GM008716 (National Institutes of Health). W.S.G. is supported by T32 DK083262. R.G.S. is supported by R01 GM084147 (National Institutes of Health) and 1BX000851 (Department of Veterans Affairs).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Short‐ and Long‐term Effects of Phenytoin Exposure on the Liver Proteome of Neonatal and Adult Mice Using SWATH‐MS Technology
Authors: Shi Jian , Piekos Stephanie C. , Chen Liming , Jing Xinyue , Zhong Xiao‐Bo , Zhu Hao‐Jie
Published: June 2021

Phenytoin, a drug listed on the World Health Organization's Model List of Essential Medicines, has been widely used for the treatment of seizures and several other indications. However, it has a narrow therapeutic index and some severe side effects, such as drug‐induced liver diseases. In addition, phenytoin is a well‐known inducer of several drug‐metabolizing enzymes and transporters expressed in the liver, which could lead to severe drug‐drug interactions. In the present study, a newly developed untargeted proteomics approach, sequential window acquisition of all theoretical mass spectra (SWATH), was utilized to investigate the short‐ and long‐term effects of phenytoin exposure on the liver proteome of neonatal and adult mice. A total of 1,780 protein groups were identified in the liver microsomes prepared from neonatal and adult mice. Markedly different liver proteome profiles were observed between newborn (day 5) and adult mice (day 60). Administration of phenytoin exhibited a greater short‐term impact on the liver proteome in neonates than that in adults, in which hepatic expression levels of 214 protein groups were significantly altered in newborn mice relative to 88 protein groups in adult mice at 24 h following an intraperitoneal injection of phenytoin (100 mg/kg). Furthermore, relative to vehicle control, phenytoin treatment resulted in significant changes of the expressions of 50 protein groups in the liver at age day 60 after the mice received a single intraperitoneal injection of phenytoin (100 mg/kg) at age day 5. Taken together, our study provides an insight into the effect of phenytoin treatment on the liver proteome, which might lead to a better understanding of the phenytoin‐induced hepatotoxicity and drug‐drug interactions.

Support or Funding Information

This study was supported by National Institutes of Health National Institute of General Medical Sciences [R01GM‐118367] (Xiao‐Bo Zhong).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Generation of an Adult Hyperbilirubinemia Model in Liver‐specific Humanized UGT1A1*6 Mice
Authors: Mennillo Elvira , Maruo Yoshihiro , Nguyen Nghia , Thérien Ariane , Chen Shujuan , Barbier Olivier , Tukey Robert H.
Published: June 2021

Bilirubin is metabolized in liver by UDP‐glucuronosyltransferase 1A1 (UGT1A1). Genetic polymorphisms associated with the UGT1A1 gene are commonly known to be responsible for increased levels of total serum bilirubin (TSB). Variants in the UGT1A1 gene promoter as well as Single Nucleotide Polymorphisms (SNPs) at both promoter and exons can result in UGT1A1 deficiency and impairment of bilirubin conjugation, leading to an icterus condition (jaundice) or hereditary unconjugated hyperbilirubinemia. Gilbert Syndrome (GS) is a common heritable condition of adult hyperbilirubinemia affecting over 10% of the population. The most common polymorphisms are the TA‐insertion in the TATA box (TA7) sequence in the promoter region of the UGT1A1 gene. In addition, a SNP at nucleotide 211G>A (G71R) in exon 1 of the UGT1A1*6 gene confers adult hyperbilirubinemia or GS. To examine the contribution of the UGT1A1*6 gene (R71) towards adult hyperbilirubinemia, a UGT1A1*6 and a UGT1A1*1 (G71) cDNA was expressed specifically in the liver of Ugt1−/− mice. Adult humanized UGT1A1*6 mice (h1A1*6HEP mice) expressed an icteric condition compared to normal TSB levels in h1A1*1HEP mice. Differences in TSB levels between h1A1*1HEP and h1A1*6HEP mice was reflected in expression levels of liver UGT1A1. The elevated TSB levels in h1A1*6HEP mice was consistent with the h1A1*6HEP mice being leaner than h1A1*1HEP mice, linking elevated TSB levels and its antioxidant potential with reduced body fat. Functional studies using liver extracts confirmed that the Km for 7‐ethyl‐10‐hydroxy‐camptothecin (SN‐38) and bilirubin glucuronidation was similar for UGT1A1*1 and UGT1A1*6, while there was a significant decrease in the Vmax associated with UGT1A1*6. The reduced Vmax is associated with reduced liver UGT1A1*6 expression and elevated TSB levels. Reduced expression of UGT1A1*6 may explain in part the contribution of elevated TSB levels in GS.

Support or Funding Information

USPHS grants GM086713 and ES010337 and grant PJT‐148611

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Engineered CaS clusters to limit breast cancer cell growth and proliferation selectively
Authors: Castro Miguel , Suarez Edu , Rivera Daniel , Adorno Wilmer , Munoz Kevin
Published: June 2021

The pH is a major difference between normal and cancer cells. Thermodynamic calculations indicate that CaS has limited dissociation in slightly basic pH, like the one found in normal cells, but dissociates easily into H2S and Ca2+ in the acidic pH found in cancer cell environments. We have studied the effect of CaS – in the size range of a few A to about 1 nm‐ in the cell cycle of breast cancer and normal fibroblasts. A single dose of CaS nanoclusters does not affect the survival and growth rate of normal fibroblasts but inhibits the proliferation rate of carcinoma cells in vitro. Human carcinoma cells treated with the CaS nanocluster dispersion exhibited a decreased ability to properly enter the cell cycle marked by a decrease in cell concentration in G0/G1 phase at 24‐ hours post treatment and then an increase in cells held in the SubG1 and G0/G1 phases up to 72‐hours post treatment. Apoptosis and necrotic channels were found to play significant roles in the death of human carcinoma exposed to the CaS nanoclusters. In contrast, any effect on normal fibroblasts appeared to be short lived and non‐detrimental. Passive uptake and intracellular pH levels of carcinoma cells are proposed to result in the observed selectivity of CaS to inhibit cancer cell proliferation with no significant effect on normal fibroblast cells. The results encourage further research with other cell lines in vitro as well as in vivo to translate this nanotechnology into clinical use.

Support or Funding Information

NIH RISE #R25GM096955NIH RISE 5R25GM088023‐02

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 hydrogel‐encapsulated T0901317 reduces atherosclerosis without effect on lipogenesis in apoE deficient mice
Authors: Li Qi
Published: June 2021

Objective

Activation of liver X receptor (LXR) by ligand induces macrophage ATP‐binding cassette transporter A1 and G1 (ABCA1/ABCG1) expression to enhance cholesterol efflux, thereby inhibiting formation of foam cells and development of atherosclerosis. However, LXR activation in hepatocytes induces fatty acid synthesis, which leads to development of fatty liver by an LXR ligand, such as T0901317. Targeting macrophage LXR selectively can avoid the adverse effects of LXR ligand on the liver. A hydrogel can be taken up by monocytes or macrophages, but not by hepatocytes. In this study, we treated apoE deficient mice with a hydrogel‐encapsulated T0901317, and determined whether the hydrogel‐encapsulated T0901317 can inhibit atherosclerosis while eliminating LXR‐induced fatty liver. We also attempted to disclose the involved molecular mechanisms.

Approach and Results

ApoE−/− mice randomly in three groups received the following treatment for 16 weeks. Group 1 (control group): mice were fed high‐fat diet (HFD); Group 2 (TF group): mice were fed HFD containing T0901317; Group 3 (THI group): mice were fed HFD while subcutaneously injected the hydrogel‐encapsulated T0901317 once every 3 days. At the end of experiment, we found that compared with control group, en face lesions were decreased by ~37.4 and ~33.17% by TF and THI, respectively. In the fibrous cap of aortic wall, the smooth muscle cell/collagen content was increased while necrotic cores, macrophages accumulation and inflammation were decreased by TF and THI. The inhibition of foam cell formation by TF and THI was attributed by LXR‐induced ABCA1/ABCG1 expression. Interestingly, compared with severe liver steatosis induced by TF, THI totally blocked T0901317‐induced fatty liver. Meanwhile, levels of serum total cholesterol, triglyceride and ALT/ALP, and liver triglyceride accumulation induced by LXR were also decreased by THI. Mechanistically, we determined that the hydrogel‐encapsulated T0901317 was taken up by monocyte/macrophages via a receptor‐mediated endocytosis, and the drug‐loaded macrophages were recruited to the lesion areas. Therefore, hepatic LXR and LXR‐targeted genes expression, such as SREBP1, ACC1 and FASN, were not activated by T0901317. As a result, hydrogel‐encapsulated T0901317 eliminated LXR ligand‐induced fatty liver.

Conclusions

Our study demonstrates that the hydrogel‐encapsulated T0901317 can reduce atherosclerosis while eliminate T0901317‐induced fatty liver, indicating the hydrogel‐encapsulated LXR ligand might be a novel therapy for atherosclerosis treatment.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Knockout of Sphingosine Kinase 1 Attenuates Kidney Damage in DOCA‐salt Mice
Authors: Lyu Bingqing , Wang Weili , Li Pin‐Lan , Li Ningjun
Published: June 2021

Sphingosine‐1‐phosphate (S1P) is a bioactive sphingolipid metabolite. S1P is produced from phosphorylation of sphingosine catalyzed by sphingosine kinases (SphK). SphK1/S1P signaling pathway is implicated in the progression of chronic kidney disease. We recently showed that knockout (KO) of SphK1 attenuated renal interstitial fibrosis in unilateral ureteral obstruction model. However, the role of SphK1/S1P pathway in the renal injury in hypertension has not yet been reported. The aim of this study was to test the hypothesis that SphK1/S1P signaling pathway mediates the kidney damage in DOCA‐salt hypertensive mice. Male wild type C57BL6 mice and Sphk1 KO mice were unilaterally nephrectomized and received a subcutaneous silicone strip containing 50 mg of deoxycorticosterone acetate (DOCA) and 1% NaCl drinking water. At the 7th week, blood pressure was measured, 24h urine collected, and kidney tissues harvested. Four groups were studied: wild type control (WT‐C), wild type DOCA (WT‐DOCA), knockout control (KO‐C) and knockout DOCA (KO‐DOCA). There was no difference in DOCA‐salt‐induced hypertension between WT and KO mice (mean arterial pressure: 135.4 ± 6.1 mmHg vs. 130.4 ± 4.7 mmHg in WT‐DOCA and KO‐DOCA mice). The urine albumin was increased in two DOCA‐salt groups and that the KO‐DOCA group showed significantly lower urine albumin than WT‐DOCA mice (0.43 ± 0.09 mg/kg/24h in WT‐C, 19.36 ± 4.84 in WT‐DOCA, 0.52 ± 0.21 in KO‐C and 3.16 ± 0.74 in KO‐DOCA, P<0.05 for WT‐DOCA vs. KO‐DOCA). The glomerulosclerosis indices were significantly lower in KO‐DOCA than WT‐DOCA in kidney tissue slides with PAS staining (0.62 ± 0.12 in WT‐C, 1.93± 0.13 in WT‐DOCA, 0.43 ± 0.05 in KO‐C, and 1.31 ± 0.11 in KO‐DOCA, p<0.05 for WT‐DOCA vs. KO‐DOCA). The protein levels of α‐smooth muscle actin (α‐SMA) in kidney tissues were upregulated in both DOCA‐salt groups but were significant lower in KO‐DOCA group than in WT‐DOCA group. The relative protein levels of α‐SMA were 1.0 ± 0.1 in WT‐C, 3.97 ± 0.55 in WT‐DOCA, 1.12 ± 0.15 in KO‐C, and 1.61 ± 0.24 in KO‐DOCA (P<0.05 for WT‐DOCA vs. KO‐DOCA). These results suggest that Sphk1/S1P signaling pathway mediates kidney damage in DOCA‐salt hypertensive mice independent of blood pressure. Manipulating Sphk1/S1P signaling pathway may be used as a therapeutic strategy in kidney damage associated with hypertension. (Supported by NIH grant DK107991 to NL and DK54927 to PL)

Support or Funding Information

NIH grant DK107991 to NL and DK54927 to PL

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Femoral artery ligation impairs skeletal muscle force production in C57Bl/6J mice
Authors: Nickell Courtney Elizabeth , Cardinal Trevor Ryan
Published: June 2021

Patients diagnosed with Peripheral Arterial Occlusive Disease (PAOD), generally caused by atherosclerotic plaque accumulation in conduit arteries, typically experience intermittent claudication, which is characterized by pain during exercise in the lower extremities due to ischemia. Perfusion distal to occluded arteries can be maintained through enlargement of natural bypass collaterals by arteriogenesis. Unfortunately, clinical trials of arteriogenesis‐ and angiogenesis‐stimulating therapies have been unsuccessful, despite clear efficacy in animal models. This may be at least partially explained by the differences in endpoints in animal models and human patients. For example, the most common assessment of ischemic revascularization in animal models is hindpaw perfusion while the most clinical endpoint is pain‐free walking distance, which depends both on vascular supply and skeletal muscle function. However, it is unclear how occlusion affects skeletal muscle performance in animal models. To better understand the natural history of the tissue response following arterial occlusion, it is necessary to determine the impact of occlusion on skeletal muscle performance using precise and quantitative methods.

Therefore, we developed a muscle force production protocol following distal femoral artery ligation (FAL). Specifically, following ligation between the epigastric and popliteal branches, we measured force production in the triceps surae, i.e. gastrocnemius and soleus, in response to single, tetanic, and fatiguing stimulations in male C57Bl/6J mice. Pilot studies indicated that force production was not different between unoperated right and left hindlimbs, with maximum twitch forces of 398.6 ± 80.5mN and 389.2 ± 41.8mN, respectively, and maximum tetanic forces of 1420.7 ± 271.4mN and 1361.7 ± 246.5mN, respectively. Following protocol development, we investigated whether forces produced at day‐7 following FAL were consistently lower than those produced by a sham‐operated hindlimb. As expected, maximum twitch force was reduced in the operated hindlimb as compared to the contralateral sham operated hindlimb, 458.95 ± 28.02 mN and 504.08 ± 28.12 mN in the operated and sham hindlimbs, respectively. Similarly, maximum tetanic force production was also reduced in the operated hindlimb, 1583.8 ± 218.2 mN, as compared to 1990.7 ± 270.7 mN in the sham. Having determined that force production is impaired at day‐7 following FAL, future studies may include characterizing the time course of force production recovery and/or investigating if force production in ligated hindlimbs can be rescued with myoblast cell transplantation.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.