bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of Ferroptosis Using UAMC‐3203 in the Post‐stroke Period Does Not Impact Cognitive Outcomes in Diabetic Rats
Authors: Edgerton Mia , Phoenix Ashley , Chandran Raghavendar , Wolf Victoria , Abdul Yasir , Jamil Sarah , Li Weiguo , Ergul Adviye 
Published: May 2022

IntroductionPost‐stroke cognitive impairment (PSCI) contributes to significant long‐term disability in stroke victims. 30% of ischemic stroke victims in the United States also have diabetes, which increases the risk of hemorrhagic transformation as well as PSCI. Ferroptosis, an iron‐induced cell death can instigate increased oxidative stress and contribute to impaired neurovascular repair leading to PSCI in diabetes. In our previous studies, we were able to identify that treating diabetic animals with an iron chelator, deferoxamine (DFX), prevents post‐stroke vasoregression and improves functional outcomes. Furthermore, DFX prevented the activation of ferroptosis in brain microvascular endothelial cells in vitro. Untreated diabetic animals experienced progressive cognitive decline while being monitored for 8 weeks. These findings led us to speculate that endothelial ferroptosis also plays a role in vasoregression and impacts cognitive outcomes post‐stroke. Therefore, this study was designed to test the hypothesis that inhibiting ferroptosis in the post‐stroke period will improve cognitive recovery in diabetic animals.MethodsAnimals were housed in reverse light cycle. 8 weeks after diabetes onset, male rats underwent 60 min middle cerebral artery occlusion (MCAO). On Day 3, after stroke injury was confirmed by MRI, animals were randomized to UAMC‐3 (2mg/kg) or vehicle treatment for 2 weeks. Sensorimotor and cognitive behavioral tests were performed during the animals’ active hours up to 8 weeks post MCAO.Results(Table 1): 60 min occlusion caused significant acute neurological deficits. There were no differences between the groups in indices measured by novel object recognition (NOR), Y‐maze and sucrose preference tests. Interestingly, step through latency in passive avoidance test (PAT) was lower in the UAMC‐3203 group.ConclusionTreatment with a ferroptosis inhibitor for 2 weeks after stroke did not impact recognition and working memory but worsened aversive learning in diabetic male rats. Unlike our previous study in which behavior tests were performed during rats’ passive hours, there was no progressive cognitive decline in untreated animals. Further evaluation of behavior testing times as well as tissue markers of neurovascular degeneration, inflammation and ferroptosis are required to determine whether molecular and cellular markers are affected by the treatment before overt changes in behavioral outcomes.
Adaptation to High‐Salt Diet Requires H,K‐ATPase Type 2 Dependent NaCl Secretion
Authors: Cheval Lydie , Lasaad Samia , Rafael Chloé , Morla Luciana , Picard Nicolas , Crambert Gilles 
Published: May 2022

IntroductionThe kidney controls the extracellular volume and the blood pressure because of its ability to excrete efficiently Na+ and Cl‐ ions. These last decades in Western countries the consumption of salt has dramatically increase and the question arises as to how the kidney adapts to this chronic situation. Up to now, the mechanisms described to excrete a large load of salt consist in the inhibition of Na+ reabsorption all along the nephron. We have recently identified a novel pathway that mediates the secretion of Na+ in the collecting duct (Morla, L. et al. 2016). This system involves A‐type intercalated cells (AIC), NKCC1 and the H,K‐ATPase type 2 (HKA2), which is known to be able to transport Na+. Here, we investigated the role of HKA2 in NaCl renal secretion, as well as the physiological relevance of this secretory pathway in vivo, in response to salt load.Material and MethodsC57BL6J mice wild type or knockout for the Atp12a gene (HKA2‐KO) were used and placed under a control (NS) or high‐Na+ (HS) diets for 3 days. Outer medullary collecting ducts (OMCD) of WT mice under NS or HS were isolated for RNAseq and RT‐PCR analysis. Type A intercalated cells were isolated by FACS (following labelling with c‐kit antibodies) from WT mice under NS or HS for RT‐PCR analysis. The mice were also placed in metabolic cages for measurements of physiological parameters (food and water intakes, urine excretion, plasma parameters). Blood pressure was assessed by tail‐cuffed measurement.ResultsThe RNA seq analysis of OMCD revealed that, as expected, genes involved in Na+ reabsorption (ENaCa, SGK) are downregulated. However, we also observed that HS diet also leads to an increased expression of the Atp12a gene that encodes for the catalytic subunit of the HKA2 both in OMCD and ICA. To understand the role of the HKA2, we placed the HKA2‐KO mice under high‐salt diet and observed that those mice exhibit a salt loosing phenotype with low blood pressure that is due to a stronger down‐regulation of NKCC2 than in WT in the same context. Therefore, the absence of HKA2 is over‐compensated by the inhibition of a reabsorption pathway.ConclusionAltogether, these results demonstrate that the secretion of Na+ in OMCD is stimulated in response to high salt intake.
Endometriosis Induces Colonic Inflammation and Reactive Gliosis in an Animal Model
Authors: Rivera‐Arce Luis A. , Rodriguez‐Cintron Ulises , Cruz Myrella L. , Appleyard Caroline B. 
Published: May 2022

IntroductionEndometriosis is a complex gynecological disorder characterized by the ectopic growth of endometrial tissue. Symptoms of endometriosis are known to impair the quality of life of patients, among these symptoms are dysmenorrhea, chronic pelvic pain, and gastrointestinal (GI) issues. These GI issues are one of the common reasons for misdiagnosis with irritable bowel syndrome (IBS), a pain syndrome characterized by abdominal pain, painful bowel movements, diarrhea, and constipation, all of which are often present in endometriosis and persist even after treatment. Enteric glial cells (EGC) are known to play a role in pain associated with IBS, and reactive gliosis has been reported in patients with IBS, but the role of EGC in endometriosis has yet to be elucidated.HypothesisWe hypothesize that endometriosis will induce reactive gliosis, with increased expression of the glial fibrillary acidic protein (GFAP) and S100B, in the myenteric plexus of colonic sections in an animal model of endometriosis.MethodsWe used 15 female Sprague Dawley rats and divided them into three groups (n=5 per group): Naïve, Sham, and Endo. Endometriosis was induced in the Endo group by suturing 4 pieces of autologous uterine tissue in the intestinal mesentery, while the Sham group received only sutures without tissue. Von Frey and Hot Plate tests were done the day before surgery, two weeks after surgery, and before sacrifice to assess pain behavior. 21 days after surgery, animals were sacrificed, and endometriosis development and colonic macroscopic inflammation were assessed. Colonic tissue was collected to assess GFAP and S100B immunoreactivity, microscopic damage, and mast cell whole‐tissue cell count.ResultsWe found Endo animals showed a decrease in allodynia threshold, but no hyperalgesia. All Endo animals developed endometriosis with an average area per vesicle of 13.89±1.43mm2, and we found Endo animals had a significant increase in the colonic macroscopic score (3.01±0.22;p<0.001) compared to Sham (0.35±0.12) and Naïve (0.47±0.20). Paraffin‐embedded colon sections stained with H&E didn’t show statistical differences in microscopic damage between groups, yet toluidine blue staining for mast cells showed a 36% increase in the mean number of cells in Endo compared to Naïve but didn’t reach statistical difference. GFAP whole‐mount immunostaining showed significantly increased integrated density in Endo (25655.00±2395.00) when compared to Sham (13558.00±719.80;p<0.001) or Naïve (9035.00±1362.00;p<0.001), while no statistical difference was found in S100B integrated density between groups.ConclusionEndometriosis can alter GI homeostasis by inducing macroscopic inflammation, reactive gliosis, and some mast cell infiltration. Taken together these suggest endometriosis can mimic IBS histopathology beyond the symptomatology, reinforcing this disease’s complexity and the need to treat it beyond the gynecological setting.
HFpEF in CKD is Associated with Elevated TNF‐α/IL‐6 Inflammatory Signaling from the Kidney
Authors: Chade Alejandro R. , Eirin Alfonso 
Published: May 2022

BackgroundChronic kidney disease (CKD) is independently associated with incident heart failure with preserved ejection fraction (HFpEF), but the underlying mechanisms remain unknown. We recently developed a translational swine model of CKD‐HF that replicates many features of human HFpEF. This study tested the hypothesis that CKD‐induced HFpEF associates with increased pro‐inflammatory cytokine signaling of renal origin.MethodsCKD and normal pigs (n=4 each) were studied for 14 weeks. Renal hemodynamics (multi‐detector CT) and cardiac morphology and function (echocardiography) were quantified in vivo. Tumor necrosis factor (TNF)‐ α and interleukin (IL)‐6 levels (ELISA) were measured in circulating, renal vein, and coronary sinus blood, and their renal and cardiac gradients quantified. Systemic TNF‐α and IL‐6 levels (Luminex) were also measured in patients with CKD and age/gender‐matched healthy volunteers (n=12 each) and correlated with biomarkers of heart failure.ResultsPigs with CKD developed hypertension, renal failure (stage 3), left ventricular (LV) hypertrophy, and abnormal LV strain and diastolic dysfunction (E/A, E/e’ ratio) with pEF, accompanied by positive renal (renal release) and negative cardiac (cardiac retention) gradients of TNF‐α and IL‐6 (Fig. 1A). Circulating TNF‐α and IL‐6 were also higher in patients with CKD compared to normal subjects (Fig. 1B), which correlated directly with ANP and NT‐proBNP levels (Fig. 1C).ConclusionsOur experimental and clinical data supports a crosstalk between the kidney and the heart in CKD, suggesting that inflammatory signaling led by TNF‐α and IL‐6 of renal origin may impose cardiac abnormalities towards development of HFpEF.
Histone H3K27 Demethylase KDM6A Regulates Cardiac Aging viaInduction of HoxC4 mediated ER Stress
Authors: Chen Kai , Zhang Bo , Sun Zhongjie 
Published: May 2022

BackgroundKDM6A mediates the removal of repressive trimethylation from H3K27me3 to activate target gene expression. It is not known, however, whether KDM6A plays a role in the regulation of cardiac aging. We hypothesized that cardiac‐specific KDM6A knockout accelerates cardiac aging.Methods and ResultsAging decreased histone demethylase KDM6A expression and activity in human cardiomyocytes. Downregulation of KDM6A expression was also found in cardiomyocytes of aged mice. The cardiomyocytes‐specific KDM6A knockout (Myh6‐KDM6A cKO) mouse model was created by using Cre‐LoxP system. KDM6A deletion didn’t cause overt cardiac remodeling and dysfunction in normal condition. However, cardiomyocytes‐specific conditional knockout of KDM6A exaggerated ISO‐induced cardiac dysfunction and cardiac remodeling. Moreover, Myh6‐KDM6A cKO mice showed early cardiac aging at 23‐month‐old, while control mice didn’t display cardiac aging until 27‐month‐old. Therefore, KDM6A deletion accelerated cardiac aging. ChiP‐Seq analysis revealed candidate transcription factors participating in KDM6A cKO‐induced cardiac aging. In particular, HoxC4 was down‐regulated in the cardiomyocytes of Myh6‐KDM6A cKO mice. Knockdown of HoxC4 gene in H9c2 cardiomyoblast cell line caused ER stress. The ER stress further resulted in oxidative stress and cardiomyocytes apoptosis in cardiomyocytes‐specific conditional knockout of KDM6A mice.ConclusionsCardiomyocytes‐specific conditional knockout of KDM6A causes cardiac aging through induction of HoxC4‐mediated ER stress. These observations suggest KDM6A a potential therapeutic target for cardiac aging.
Emerald Jewel Wasp Venom Adenosine Deaminase Antagonizes Purinergic Signaling in the Cockroach Brain
Authors: Connolly Maize , Theisen Rachel , Arvidson Ryan 
Published: May 2022

Ampulex compressa injects venom directly into the cerebral ganglia of the cockroach (Periplaneta americana) to induce a reversable, but long‐term compliant state of hypokinesia, marked by an increase in escape threshold and a decrease in spontaneous walking. The venom is not paralytic as stung cockroaches can still stand, groom, eat and drink, and can even run if aggressively provoked. The venom is also not necrotic or cytotoxic, and despite its long‐lasting duration the venom’s effects are reversible, as the escape response returns to normal after several days. Previous work involved generation of a comprehensive venom apparatus transcriptome and venom proteome or “venome” and described several interesting signaling pathways that appear to be targeted by venom components. This includes adenosine deaminase (ADA), which may act to antagonize purinergic signaling in the brain.The substrates of venom ADA, adenosine (Ado) and adenosine monophosphate (AMP), are also the activating ligands of the cockroach adenosine receptor (AdoR). We hypothesize that the presence of ADA at the site of injection can reduce cell damage‐induced signaling by clearing Ado and AMP released from compromised cells. We have cloned, expressed, and characterized both venom ADA and cockroach brain AdoR and show that the presence of venom ADA can attenuate Ado induced calcium signaling in vitro. Antagonizing ADA or AdoR in vivo did not alter locomotory behavior. Thus, the action of ADA in the venom may be more neuroprotective than inhibitory of specific behaviors as is the case with other venom components.
Does CADM1 Immunohistochemistry Distinguish Mycosis Fungoides from its Closest Mimickers?
Authors: Haghayeghi Koorosh , LeBlanc Robert E. 
Published: May 2022

BackgroundImmunohistochemistry (IHC) sometimes fails to distinguish clinically ambiguous cases of evolving patch‐stage mycosis fungoides (MF) from relatively common, non‐neoplastic inflammatory skin diseases (ISD), particularly spongiotic dermatitis (SD), psoriasis (PSO), and pityriasis lichenoides (PL). CADM1 (cell adhesion molecule 1) is a tumor suppressor gene implicated in the pathogeneses of solid skin tumors, visceral malignancies, and peripheral T‐cell lymphomas. Recent data suggest that CADM1 IHC expression could help distinguish early‐stage MF from ISD.DesignThis pilot study sought to determine whether CADM1 IHC distinguishes MF from ISD in clinically‐annotated cases from our archives. MF (n=14), PL (n=10), PSO (n=10), and SD (n=10) were stained with CADM1. Using appropriate controls and antibody optimization, two pathologists established consensus scores in each case for percentage of lymphocytes staining with CADM1 and CADM1 staining intensity. The localization of CADM1‐positive cells (epidermis including junction and dermis) was also noted.ResultsThere were no differences in CADM1 expression pattern or intensity between groups. All cases showed moderate to strong nuclear CADM1 expression in ≥75% of lymphocytes in both the dermis and epidermis. Some intraepidermal lymphocyte nuclei were challenging to detect on CADM1‐stained slides given uniform moderate‐intensity staining of background keratinocyte nuclei.ConclusionThe challenge of distinguishing evolving patch‐stage MF from ISD facilitates demand for novel and readily available diagnostic tests. Although recent data have suggested that differential expression of CADM1 distinguishes MF from ISD, there was no difference in lymphocyte CADM1 expression and distribution between any of the groups included in our pilot study. Furthermore, evaluation of CADM1 expression in intraepidermal lymphocytes was challenging given background keratinocyte expression.
The Loss of Endothelial Glycocalyx in Diabetic Retinopathy
Authors: Kaur Gaganpreet , Song Yuefan , Xia Ke , Cruz‐Topete Diana , McCarthy Kevin , Zhang Fuming , Linhardt Robert , Harris Norman 
Published: May 2022

IntroductionThe vascular endothelium is lined with the endothelial glycocalyx, composed of core proteins and glycosaminoglycans (GAGs). The loss of glycocalyx can cause endothelial injury and could contribute to the progression of retinopathy. The glycocalyx thickness has been shown to decrease in the diabetic retina; however, changes in individual components of the retinal glycocalyx have yet to be determined. Therefore, the present study examined hyperglycemia‐induced changes in retinal glycocalyx components.MethodsType‐1 diabetic rats and primary rat retinal microvascular endothelial cells (RRMECs) exposed to high glucose were used to mimic diabetes. Western blots and qRT‐PCR were used to analyze the expression of core proteins. Further, GAG composition was studied using LC‐MS.ResultsBoth retinal endothelial cells and rat retina exhibited a significant decrease in mRNA transcripts and protein levels of syndecan‐3 under hyperglycemic condition, whereas syndecan‐1 expression was significantly increased. Further, we observed a substantial loss of glypican‐1 in RRMECs with high glucose, but a significant increase in the retina of diabetic rats. RRMECs in high glucose exhibited a significant decrease in chondroitin sulfate (CS) and hyaluronic acid (HA), but no change in heparan sulfate (HS) levels. In addition, 4‐O‐sulfated CS residues and 2‐O‐N‐sulfated HS residues were reduced, whereas N‐sulfated and 6‐O‐N‐sulfated HS residues were increased in retinal endothelial cells treated with high glucose. Lastly, we observed a significant increase in media levels of syndecan‐1, HA, HS, and CS, indicating increased shedding of glycocalyx components under hyperglycemia.ConclusionsOur findings suggest that the loss of endothelial glycocalyx in the diabetic retina could be a result of a loss of the proteoglycan syndecan‐3 (and possibly glypican‐1) as well as the GAGs CS and HA.
Age‐dependent differences in the ventilator‐induced diaphragm dysfunction (VIDD): A multi‐omics study
Authors: Wen Ya , Lyu Qiong , Zhang Xiang , Larsson Lars 
Published: May 2022

Introduction & ObjectiveMechanical ventilation is a life‐saving intervention in patients with respiratory failure. Controlled mechanical ventilation (CMV) is the mode of mechanical ventilation used in patients with an insufficient central respiratory drive and frequently used in COVID‐19 patients ventilated in the prone position. However, CMV is also associated with negative consequences for the major respiratory muscle the diaphragm, i.e., the ventilator induced diaphragm dysfunction (VIDD). VIDD is the major factor underlying the delayed weaning from the ventilator especially in old intensive care unit (ICU) patients. The age‐related difference in mortality rate has become a major concern during the COVID‐19 pandemic. The current experimental study was undertaken to improve our understanding the mechanisms underlying age‐related differences in the response to CMV using a unique experimental ICU model allowing long‐term studies in a rat exposed CMV.Materials and MethodsYoung (7‐8 months) and old (28‐32 months) female F344 BN hybrid rats were exposed to the ICU condition for 5 days (anaesthetized, mechanically ventilated, and pharmacologically paralyzed by post‐synaptic blockade of the neuromuscular transmission). Transcriptomics (RNA‐Seq) and proteomics (Olink) analyses of the diaphragm and proteomics analysis of plasma were conducted to investigate the molecular differences between young and old rats exposed to the ICU condition.ResultsAccording to multi‐omics analyses, significant differences were observed in the diaphragm between young and old rats in response to ICU condition. In the diaphragm of young rats, CMV resulted in decreases in the biological processes related to energy metabolism and production as indicated by the downregulation of pathway like “glycolysis/gluconeogenesis”, “fatty acid metabolism”, “TCA cycle”, and “oxidative phosphorylation”, etc. On the other hand, in the diaphragm of old rats, dramatic immune and inflammatory responses were observed after CMV, as reflected in the upregulation of “Cytokine‐cytokine receptor interaction”, “TNF signaling pathway”, “IL‐17 signaling pathway”, etc. Transcriptomic results were consistent with plasma proteomics analyses, i.e., CMV induced a higher level of inflammatory factors and cytokines/chemokines in old rats.ConclusionsAfter 5‐days exposure to CMV, our multi‐omics study demonstrated age‐related inflammatory response to long‐term mechanical ventilation.Significance/ImplicationThus, CMV should be cautiously used and restricted to as short duration as possible, especially in the old patients. In the current COVID‐19 pandemic, the dramatically increased mortality in old ICU patients with COVID‐19‐associated hyperinflammation and cytokine storm need not only reflect the viral infection but may also be associated with the VIDD and hyperinflammatory responses induced by CMV per se.
The Regulatory Role of miR‐21 in Coronary Microcirculation
Authors: Juguilon Cody , Wang Yang , Wang Zhiyuan , Enrick Molly , Gadd James , Clark Alyssa , Chilian William , Yin Liya 
Published: May 2022

IntroductionCoronary microvascular dysfunction is characterized by impaired endothelial‐dependent vasodilation. These impairments are seen in diabetic cardiomyopathy (DCM), coronary artery disease (CAD) and ischemia with non‐obstructive coronary artery (INOCA), Takotsubo cardiomyopathy, myocardial infarction with non‐obstructive coronary artery disease (MINOCA), and heart failure with preserved ejection fraction (HFpEF), but detailed mechanisms have yet to be elucidated.MethodsmicroRNA‐21 (miR‐21) global and conditional knockout mice were used to study how miR‐21 regulates coronary microcirculation in pathological conditions like DCM. Both genetic (db/db) and diet‐induced diabetic models were used. Coronary arteries were isolated, and endothelial‐dependent vasodilation was assessed using myography (DMT). In vivo myocardial blood flow (MBF) under stress was measured by contrast echocardiography or doppler after the treatment with different dosages of norepinephrine. Quantitative polymerase chain reaction (qPCR) was performed for gene expression analysis. Trichrome staining and histology were performed for structural changes of the hearts.ResultsOur preliminary data show that miR‐21 is upregulated in DCM and the deficiency of miR‐21 restores the endothelial‐dependent vasodilation in isolated diabetic coronary arterioles and coronary blood flow under stress in DCM through the mechanism that miR‐21 prevents the mediator of coronary vasodilation switching from NO to H2O2 in diabetes.ConclusionsmiR‐21 regulates microvascular dysfunction in DCM. Further genetic profiling will elucidate the pathways and mechanisms converging with miR‐21 to regulate microvascular function.