bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of PC‐NSAIDs to protect gingival cells from injury due to cytotoxic agents
Authors: Lichtenberger Lenard M. , Dial Elizabeth J. , Fang Dexing 
Published: November 2023

BackgroundOral mucositis is perhaps the most severe oral side effect of cancer treatment and consists of painful inflammation and ulceration of the mucous membranes lining the oral cavity. Oral mucositis can make it difficult to eat and speak, thereby limiting the dosage and/or duration of radiation/chemotherapy, and require treatment modifications and compromises to prognosis. Antibiotic/anti‐fungal agents have limited efficacy at preventing or treating oral mucositis, also a number of “Magic Mouthwashes” that contain a variety of ingredients have not been shown to be effective, although work continues in this direction. Some other agents, such as the calcium phosphate mouth rinse Caphosol®, have shown efficacy in small clinical studies, but not been widely tested yet. Finally, due to the irritating potential on the mucosal surface any rinses with alcohol are not recommended. Clearly, there is a great unmet need for a new approach to prevent and/or treat mucositis.Objective/AimMacrophages and gingival fibroblast cells are the major cells in mouth responding to mucositis. The goal of this study is to use an in vitro cell culture system of murine gingival cells (ESK‐1) to screen novel formulations of phosphatidycholine (PC) alone and PC‐complexed to the non‐steroidal anti‐inflammatory drug (NSAID), ibuprofen (IBU) for protection of the cells from clinically relevant challenges. The hypothesis is that PC, IBU and IBU‐PC will protect the gingival cells from injury due to cytotoxic agents such as, sodium dodecyl sulfate (SDS), lyso‐PC (LPC), and lipopolysaccharde (LPS), an endotoxin released by toxic bacteria due to the surface protective and/or anti‐inflammatory action of these agents.MethodsThe murine gingival ESK‐1 cells were cultured in the presence of injurious chemicals, cytotoxins and PC, IBU, or IBU‐PC was added at a range of doses. The damaging agents/toxins conditions included SDS ‐ a detergent commonly present in toothpaste, Lyso‐PC ‐ a metabolite of PC, and LPS ‐ an endotoxin released by toxic bacteria. The MTT assay was used to determine the number of cells protected and the LDH (lactate dehydrogenase) assay was used to test the cells damaged.Results & ConclusionThe cell number, release of LDH, the cytosolic protein into the culture medium as indices of cytotoxicity, demonstrated that chemical‐induced injury to the gingival cells could be significantly reduced, if not prevented, by adding IBU‐PC or PC alone to the culture medium compared to IBU alone. Interestingly, LPS‐induced release of the pro‐inflammatory cytokine IL‐6 into the medium was significantly inhibited by IBU‐PC or IBU alone compared to PC alone, indicating the efficacy of IBU and IBU‐PC to block the release of this inflammatory mediator. Taken together, IBU‐PC could be an ideal drug in combined benefit of gingival cell protection and inhibiting inflammation of the oral mucosa. Supported by a NIH NCI grant, 1R21CA202751.Support or Funding InformationSupported by a NIH NCI grant, 1R21CA202751
Defining the role of different MAPK signaling pathways following APAP and AMAP exposure
Authors: Nguyen Ngoc Uy , Valeu Lynn , Stamper Brendan 
Published: November 2023

ObjectiveThe objective of this study was to better characterize the role of mitogen‐activated protein kinase (MAPK) activation in acetaminophen (APAP) toxicity using structure‐toxicity relationships with acetyl‐m‐aminophenol (AMAP). Both c‐Jun N‐terminal kinase (JNK) and extracellular signal regulated kinase (ERK) signaling were investigated.MethodsDose‐dependent toxicity assays were performed in an in vitro liver model (TGF‐a transgenic mouse hepatocytes; TAMH) to determine non‐toxic concentrations of the following MAPK modulators: ERK inhibitor (FR‐180204), ERK inducer (Menadione – MEN), JNK inhibitor (JNK‐IN‐8, SP600125, Leflunomide ‐ LEF), and JNK inducer (cycloheximide – CHX). Protein immunoblotting was then used to confirm MAPK activation. Pretreatment with modulators was then coupled to toxicity assays with APAP and AMAP to determine their effect on cell viability. Quantification of additional protein biomarkers related to MAPK signaling was also assessed in an attempt to elucidate mechanistic details.ResultsMeasured non‐toxic concentrations of modulators were as follows: 10mm for FR‐190204, 2mM for MEN, 2nM for CHX, 50mM for LEF, 1mM for JNK‐IN‐8, and 1mM for SP600125. Time‐based immunoblotting results demonstrated peak modulation by MEN, LEF, JNK‐IN‐8, and SP600125 to occur after two hours, whereas peak CHX‐mediated JNK induction occurred after 24 hours. Induction of either JNK or ERK decreased cell viability when dosed in conjunction with APAP, with no observable effect on AMAP.ConclusionPrevious studies have established the role of JNK during APAP‐induced hepatocellular injury, whereas the role of ERK is poorly defined. As expected, when dosed with APAP, inducing JNK promoted cell death; however, the induction of ERK also promoted cell death. AMAP, a relatively non‐toxic regioisomer of AMAP was not affected by pretreatment with MAPK modulators suggesting that additional work is needed to clarify the role of ERK in APAP‐induced injury.Support or Funding InformationSource of Research Support: This work was supported by a Pacific University Research Incentive Grant
MicroRNA‐122 protects mice and human hepatocytes from acetaminophen toxicity by regulating CYP1A2 and CYP2E1 expression
Authors: Chowdhary Vivek , Luna Joseph , Teng Kun‐yu , James Laura , Lee William M , Ghoshal Kalpana 
Published: November 2023

Although acetaminophen (APAP) is a widely used anti‐pyretic and analgesic, and generally a safe drug, it is a dose‐related toxin in all mammals and causes ~1000 cases of acute liver failure (ALF) annually in the United States. Acetaminophen (APAP) toxicity is a major cause for acute liver failure (ALF) and N‐acetylcysteine (NAC) remains only useful antidote for APAP toxicity till now. MicroRNA‐122 (miR‐122) is a liver‐specific conserved microRNA that regulates metabolic homeostasis and Hepatitis B and C virus replication. It is abundantly expressed in adult mammals and regulates expression levels of many targets in hepatic cells. Although circulating miR‐122 is a sensitive biomarker of ALF, role of this most abundant liver specific miRNA in ALF has not been elucidated. Here, we show that miR‐122 is down regulated in liver biopsies of ALF patients compared to non‐ALF biopsies and in the livers of mice treated with APAP. A dramatic decrease in primary‐miR‐122 expression occurs upon APAP overdose in mice with suppression of its two key trans‐activators, HNF4a and HNF6. More importantly, mortality rate of liver‐specific miR‐122 knockout (LKO) (Mir122loxP/loxP; Alb‐Cre) mice was higher (P<0.0001) than that of the control (Mir122loxP/loxP) mice when injected intraperitoneally with APAP (500mg/kg), which is not lethal to the control mice. Higher basal level of CYP2E1 and CYP1A2 that convert APAP to highly reactive N‐acetyl‐p‐benzoquinone imine (NAPQI) are contributing factors to the sensitivity of LKO mice to APAP overdose. CYP2E1 was upregulated at the protein level whereas Cyp1a2 gene was activated in LKO livers due to upregulation of its key trans‐activator, AHR and the co‐activator MED1. Interestingly, binding of miR‐122 with an intronic site of Ahr primary transcript (hnRNA) and with a 3′‐UTR site of Med1 was identified in the control livers by Argonaute‐HITS‐CLIP analysis, which was validated by the luciferase reporter assay. The loss of these interactions resulted in derepression of Ahr hnRNA and Med1 mRNAs in Mir‐122−/− livers. miR‐122 also suppressed human AHR and MED1 expression by interacting with their 3′‐UTRs. Notably, miR‐122 depletion sensitized differentiated human HepaRG cells to APAP toxicity that correlated with upregulation of AHR, MED1 and CYP1A2 expression. Collectively, these results suggest a critical role of miR‐122 in acetaminophen metabolism and implicate its therapeutic potential in ALF.Support or Funding InformationThis study is supported, in part, by grants R01A193244, R01086978 from NIH and Pelotonia Idea grant.
Paradoxical Upregulation of Macrophage CXCR7 by Myeloid‐specific Gene Editing in Mice
Authors: Wang Chuan , Alqurashi Thamer , Shen Jianzhong 
Published: November 2023

ObjectiveWe previously reported that the chemokine receptor CXCR7 is induced during monocyte‐to‐macrophage differentiation in vitro, leading to a switch of SDF‐1 signaling profile and increased macrophage migration/phagocytosis. However, the role of monocyte/macrophage CXCR7 in vivo remains unknown.Methods and ResultsThe CXCR7‐floxed mice (CXCR7‐floxed+/+) were crossed with the LyzM‐Cre mice (LyzM‐Cre+/+) to obtain myeloid‐specific deletion of CXCR7 in vivo. Unlike the systemic knockouts, this new line was viable, fertile, and did not show obvious gross physical or behavioral abnormalities. However, we found significantly higher number of periphery blood monocytes in CXCR7‐floxed+/+/LyzM‐Cre+/+ mice compared with CXCR7‐floxed+/+/LyzM‐Cre−/− mice, indicating a role of CXCR7 in monocytosis. Further study indicated that this phenotype cannot be solely attributed to the disruption of LyzM gene by Cre insertion or the inclusion of floxed sequence to CXCR7 gene. Surprisingly, CXCR7 mRNA and protein expression were dramatically upregulated in CXCR7‐floxed+/+/LyzM‐Cre+/+ mice compared with littermate CXCR7‐floxed+/+/LyzM‐Cre−/− mice, both in bone marrow‐derived macrophages and in peritoneal macrophages, suggesting a potential compensatory mechanism. This is supported by our finding that PCR mapping indicated partial deletion of myeloid CXCR7 gene via the Cre‐LoxP approach and that adenovirus delivery of the Cre gene into CXCR7‐floxed macrophages in vitro mimicked CXCR7 induction in vivo. In addition, in the mouse peritonitis model, injection of thioglycolate led to more macrophage infiltration into the peritoneum in CXCR7‐floxed+/+/LyzM‐Cre+/+ mice.ConclusionsMyeloid‐specific manipulation of CXCR7 gene by using Cre‐LoxP method leads to compensatory upregulation of CXCR7 in monocytes/macrophages, which is associated with increased monocytosis and cell migration in vivo.Support or Funding InformationAHA national SDG award
Ubiquitin Proteasome System Inhibition Reduces LRP1 Protein Levels in HepG2 Cells
Authors: Rondón‐Ortiz Alejandro Nico , Gonzales‐Urday Ana Lucia , Martínez‐Málaga Jimena A. , Carr Olivia C. , Maher Timothy J. , Pino‐Figueroa Alejandro 
Published: November 2023

Ubiquitin proteasome system (UPS) is a conserved proteolytic process involved in the degradation of damaged and misfolded proteins. Proteins undergoing this event must be conjugated to one or more monomers of ubiquitin, a process known as ubiquitination. UPS inhibition produces protein aggregation which causes cytotoxicity. Low‐density lipoprotein receptor‐related protein‐1 (LRP1) is a transmembrane receptor composed of two subunits, α (515 kDa) and β (85 kDa) located at extracellular and intracellular levels, respectively. This receptor is involved in the uptake of molecules via clathrin for either transcellular transport or lysosomal degradation. UPS inhibition by MG132 increases LRP1 half‐life in HepG2 cells as reported in the literature. Conversely, protein aggregation might have a role in LRP1 reduction.The purpose of this research was to demonstrate that MG132, a UPS inhibitor, induces cytotoxicity, protein aggregation and reduces LRP1 protein levels in HepG2 cells. These cells were treated for 24 hours with different concentrations of MG132 (0.03 – 3.0 μM) reducing cell viability in a concentration‐dependent manner determined by 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS assay). Whole cell lysates were extracted and separated on SDS‐PAGE. Immunoblotting was used to determine K48‐linked ubiquitinated proteins and LRP1. MG132 at 0.3 and 1 μM increased K48‐linked ubiquitinated proteins and reduced LRP1 protein levels in a concentration‐dependent manner.These results suggest that UPS inhibition promotes protein aggregation, which can be the cause of cytotoxicity. In addition, MG132 reduced LRP1 protein levels which might be a consequence of protein aggregate accumulation. Further experiments are required to elucidate the LRP1 reduction mechanism when UPS is inhibited.
Prenatal exposure to cyclosporine impair cardiac function in neonates
Authors: El‐Mas Mahmoud , El‐Bassossy Hany , Awan Zuheir 
Published: November 2023

Although cardiotoxicity has been recognized as an adverse effect of cyclosporine A (CSA), limited or no information exists regarding effect of prenatal exposure on the cardiac function in the newborn. We tested the hypothesis that prenatal exposure to CSA impair cardiac function in neonates.Prenatal exposure to CSA (15 mg/kg/day sc) from the day 6 of pregnancy till delivery resulted in a sex‐dependent impairment in ventricular systolic functions in neonates as reflected by a tenfold decrease in ESPVR slope in male neonates while two‐fold decreases in female neonates. In addition, prenatal exposure to CSA led to a significant rise in systolic and diastolic blood pressure in male neonates but not in female neonates. prenatal exposure to CSA caused significant increases in JT interval and tendency to increase in QTc interval in male neonates, but not in females, indicating LV ischemia and arrhythmogenesis. On the other hand, prenatal exposure to CSA resulted in a delay in AV conduction in female neonates, but not in male, rats as reflected by the significant prolongation in P duration and tendency to increase in PR intervalIn conclusion, In summary, the current study establishes the importance of the rat sex in defining the magnitude of cardiotoxicity due to CSA prenatal exposureSupport or Funding InformationThis work was funded by the Deanship of Scientific Research (DSR), King Abdulaziz University, Jeddah, under grant No. (166 ‐ 110 ‐ D1437).
Probable renin/prorrenin receptor fetal programming in the offspring from rats with pregnancy‐induced hypertension
Authors: Cabrera‐Becerra Sandra Edith , Anguiano‐Robledo Liliana , Gallardo‐Ortiz Itzell A. , Lopez‐Sanchez Pedro 
Published: November 2023

It has been described that fetal development is related to health state in adulthood. Barker et. al. have hypothesized that the prevalence of some diseases in adults (as hypertension) are related to intrauterine development, calling this phenomenon as “fetal programming”. Renin angiotensin aldosterone system (RAAS) participates in blood pressure regulation. This system has become more complex due to the finding of new components, such as the prorrenin/renin receptor (PRR), and it is described that some of them might be submitted to fetal programming. The aim of this work was to determine PRR expression in preterm (3rd week of pregnancy), and 1, 4 and 12 weeks old offspring from normal pregnant and pregnancy‐induced hypertensive (PIH) rats. We measured blood pressure, length and body weight and determined PRR and PLZF expression by immunoblot on preterm (3rd. week gestation),1, 4 and 12 weeks old groups. We found lower body weight and length in pups at the third week of gestation, and in 1 and 4 weeks old offspring from PIH. Blood pressure was higher in offspring from PIH rats at 12 weeks. We also found PRR and PLZF expression increased in those groups. These results suggest that PRR might be programmed in fetal stage, and could play a role in development of cardiovascular diseaseSupport or Funding InformationSIP 20150467 and 20160667
Role of lipocalin‐2 in dietary and aldosterone‐induced renal damages
Authors: Sun Kiwi Wai Yan , Yang Kangmin , Luo Cuiting , Félétou Michel , Villeneuve Nicole , Vanhoutte Paul M , Xu Aimin , Wang Yu 
Published: November 2023

Study objectiveBoth high fat diet and aldosterone challenge can cause significant renal damage, either as a primary cause or as co‐morbidities with other diseases (e.g. cardiovascular diseases and diabetes). Angiotensin‐converting‐enzyme inhibitor (ACEi) and mineralocorticoid receptor inhibitor (MRi; e.g. spinronolactone and eplerenone) are commonly used in these patients. However their mortality is still high and such treatments are not suitable for all patients. Thus, a new therapeutic approach is warranted for renal damages. Herein, we hypothesize that lipocalin‐2 plays a detrimental role in both dietary and aldosterone‐induced renal injuries, and thus could be a novel drug target for therapeutic development.MethodsWild‐type (WT), lipocalin‐2 knockout (LKO) and lipocalin‐2 tissue specific knockout in adipose tissue (ADN‐Cre) mice were subjected to eight weeks of high fat diet treatment or to nephrectomy followed by four weeks of aldosterone/nephrectomy/1% saline treatment (ANS). MRi (either spinronolactone or eplerenone) was administered in ANS‐treated groups to compare its efficacy versus lipocalin‐2 deficiency mice in terms of anti‐hypertensive effect, renal damages and fibrosis, and small resistance artery stiffness and tone. Adipose tissue of all mice was also collected for further analysis.ResultsCompared to MRi‐treated wildtype mice, LKO and ADN‐Cre mice showed a better protection against ANS‐induced renal damages, as evidenced by: (1) attenuation of tissue fibrosis, (2) decreased infiltration of macrophages, (3) reduced levels of molecular injury markers, such as kidney injury marker (KIM)‐1, monocyte chemoattractant protein (MCP)‐1, transforming growth factor (TFG)‐β and collagen I. In addition, LKO and ADN‐Cre mice treated with high fat diet showed significant protection from renal damages and reduced adipose tissue fibrosis when compared to controls. As regards small resistance artery stiffness and compliance, ADN‐Cre mice were protected against ANS and high fat diet challenge.ConclusionLipocalin‐2 is involved in the development of the metabolic‐renal syndrome. Targeting its expression and/or signalling pathways could be a novel therapeutic approach to combat these medical complications.Support or Funding InformationThis work was supported by the grants from Hong Kong Research Grant Council [General Research (17121714, HKU 763312 and HKU 780613M), Collaborative Research Fund (C705514G)] and an educational grant from Les Laboratories Servier (RS100503).
An epoxyeicosatrienoic acid analog treated adriamycin‐induced proteinuria and renal fibrosis
Authors: Khan Md Abdul Hye , Goldspink Paul , Imig John D , Falck John R 
Published: November 2023

Chronic Kidney Disease (CKD) is a global health concern affecting 16% of the population and an estimated 33 million Americans. Persistent proteinuria and renal fibrosis are the two major contributors in CKD and its progression to end‐stage renal disease. Epoxyeicosatrienoic acid (EET), a CYP epoxygenase metabolite of arachidonic acid, and the EET analogs demonstrated diverse biological actions including renal protection. The aim of this study was to determine if EET‐A, an EET analog, could be used as a treatment for CKD using an adriamycin‐induced kidney injury model. Adriamycin (ADR; 10mg/kg i.v.) was administered to two groups (n=8/group) of male Balb/c mice (8–10 weeks old) whilst the control received saline (i.v.). Urine samples were collected 5 days after ADR injection and 24hr urinary protein excretion was determined. The ADR mice (15.0±4 mg/d) developed elevated proteinuria 5 days after ADR injection compared to control (2.4±0.4 mg/d). After confirming proteinuria 5 days post ADR administration, the ADR mice were treated with vehicle or EET‐A (10mg/kg in drinking water) for two weeks. In ADR mice, EET‐A treatment was able to prevent further proteinuria (11±2 mg/d) compared to the vehicle (25±4 mg/d). EET‐A treatment also reduced renal fibrosis in ADR mice by lowering the collagen positive fibrotic area (1.0±0.1%) compared to vehicle (4.0±1.2%). Overall, the data demonstrated that EET‐A treated ADR‐induced proteinuria and renal fibrosis, and has potential for CKD treatment.Support or Funding InformationResearch Starter Grant to Md. AH Khan from PhRMA Foundation, USA
CYP4A/CYP2C Modulation of the Interaction of Calcium Channel Blockers with Cyclosporine on EDHF‐Mediated Renal Vasodilations in Rats
Authors: Hammoud Safaa H , Omar Amal G , Eid Assaad A , El‐Mas Mahmoud M 
Published: November 2023

The endothelium‐derived hyperpolarizing factor (EDHF) is a back‐up mechanism that compensates for reduced nitric oxide (NO)/prodtanoids bioavailability. Here we investigated whether (i) cyclosporine (CSA, immunosuppressant drug) upregulates EDHF‐dependent renal vasodilations through altering CYP4A/CYP2C signaling, and (ii) calcium channel blockers modulate the CSA/EDHF/CYP interaction. Rats were treated with CSA, verapamil, nifedipine, or their combination for 7 days. Systolic blood pressure (SBP) was measured by Tail‐cuff plethysmography. Kidneys were then isolated, perfused with physiological solution containing L‐NAME (NOS inhibitor) and diclofenac (cyclooxygenase inhibitor), and preconstricted with phenylephrine. Carbachol vasodilations were established before and after the inhibition of 20‐HETE or EETs. CSA (25 mg.kg−1.day−1 for 7 days) increased SBP and augmented carbachol renal vasodilations. The concurrent administration of verapmil (2 mg.kg−1.day−1) or nifedipine (3 mg.kg−1.day−1) abolished the hypertensive effect of CSA and produced divergent effects on carbachol vasodilations (increases versus decreases). The infusion of MSPPOH (EETs inhibitor) reduced carbachol vasodilations in perfused kidneys of all rat groups, suggesting the importance of EETs in these responses. Moreover, HETE‐20 inhibition by HET0016 increased carbachol vasodilations in control rats, an effect that disappeared in CSA‐treated, and reappeared in rats treated with CSA/verapamil or CSA/nifedipine. The renal protein expression of CYP2C and CYP4A as well as their vasoactive products (EETs and 20‐HETE) were increased in CSA‐treated rats. Whereas the CYP2C/EETs effects of CSA were abolished by verapamil and intensified by nifedipine, the CYP4A/20‐HETE effects were reduced by verapamil only. Overall, CYP2C/EETs and CYP4a/20‐HETE pathways variably modulate the contracting effects of nifedipine and verpamil on CSA‐evoked facilitation of EDHF‐dependent renal vasodilations. These effects may not be related to changes in SBP.Support or Funding InformationSupported by the Lebanese National Council for Scientific Research (CNRS) Grant Award for Lebanese graduates (Awardee: Safaa Hammoud).