bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sickle cell disease induced hepatopathophysiology involves reduced bile acid synthesis and transport
Authors: Pradhan‐Sundd Tirthadipa , Vats Ravi , Singh Sucha , Koshar Karis , Tutuncuoglu Egemen , Nejak‐Bowen Kari , Sundd Prithu , Monga Satdarshan P 
Published: June 2021

Sickle cell disease (SCD) is an autosomal‐recessive‐genetic disorder that affects millions of people worldwide. Although hepatic crisis affects 10–40% of hospitalized SCD patients and can progress to fatal liver failure, the current treatment is primarily supportive and the molecular pathophysiology remains largely unknown. We found that transgenic, humanized SCD mice developed liver injury with age, manifested by progressive hepatomegaly, hyperbilirubinemia, iron accumulation, dyslipidemia, fibrosis, increased inflammation and necrosis. Remarkably, real‐time in‐vivo imaging of liver in live mice revealed presence of sinusoidal ischemia, impairment of bile‐secretion, and accumulation of bile‐acids in SCD but not control mice. Interestingly, impaired bile secretion was associated with the loss of apical bile‐transporters and bile‐acid converting enzymes as well as gain of efflux bile‐transporters. These findings are the first to identify that impaired bile acid synthesis and misexpression of bile transporters in SCD, contribute to liver injury by promoting intrahepatic bile accumulation and impaired bile secretion into bile canaliculi. Improved understanding of these processes could potentially benefit the development of new therapies to treat sickle cell hepatic crisis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Post‐Natal Growth and Liver Immune Homeostasis are Post‐Transcriptionally Regulated by Tristetraprolin Family of RNA Binding Proteins
Authors: Patial Sonika , Blackshear Perry 
Published: June 2021

Tristetraprolin family of RNA binding proteins, including tristetraprolin (TTP), zinc finger protein 36 like 1 (ZFP36L1), and zinc finger protein 36 like 2 (ZFP36L2) regulate mRNA levels by binding to AU‐rich elements on the 3′ untranslated regions of specific mRNAs and enhancing their turnover. While the three proteins behave similarly under biochemical conditions, they appear to have diverse physiological functions. This is apparent from the global deletion of the three proteins in mice. While the global loss of TTP results in systemic inflammatory syndrome, global loss of ZFP36L1 is embryonic lethal, and the global loss of ZFP36L2 results in early post‐natal mortality. Here, we sought to understand the physiological functions of the three proteins in liver by conditional liver specific deletion using Cre‐loxP technology. While the independent deletion of the three proteins did not result in any aberrant phenotype, simultaneous deletion resulted in post‐natal growth defect, chronic active portal hepatitis, and bile duct hyperplasia. Transcriptomic analysis showed small numbers of gene expression changes in the liver of ZFP36L1 knockout, no changes in the livers of TTP and ZFP36L2 knockout, and many changes in the liver of mice deleted of the three proteins simultaneously. The changes included perturbation of genes that regulate post‐natal growth and several inflammatory mediators, some of which are novel potential liver‐specific targets of TTP family proteins. Interestingly, one allele of either ZFP36L1 or ZFP36L2 was sufficient to protect the mice from developing this phenotype. However, one allele of TTP was only partially protective. In summary, our results suggest that post‐transcriptional regulation of gene expression plays an important role in post‐natal growth and liver immune homeostasis and that TTP family proteins regulate these physiological functions in both specific and redundant manner.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Improved Method to Capture a Broader Array of High Density Lipoprotein Particles Including Those of Intestinal Origin
Authors: Kang Jea Woo , Rhodes Christopher H. , Houts Hannah , Zivkovic Angela M. 
Published: June 2021

The isolation of high density lipoprotein (HDL) particles by density‐based methods using a density range of 1.063–1.21 g/mL is considered to be the gold standard. However, it is known that several HDL subclasses, including discoidal/nascent HDL particles, as well as ApoE‐containing, and intestinally derived ApoA‐IV‐containing HDL particles, are found at a higher density of up to 1.25 g/mL. In addition, using density‐based approaches alone it is not possible to purify HDL particles from a range of other particles such as extracellular vesicles, which overlap in density but not size. In this study, a novel method was optimized to isolate a broader array of HDL particles in a purified fraction that minimizes contamination by extracellular vesicles. The method utilizes density‐based sequential flotation ultracentrifugation followed by size exclusion chromatography with an FPLC system to isolate HDL fractions of different sizes, which were then analyzed for their cholesterol efflux capacity and protein content. The results demonstrate a higher yield of HDL and an increased level of HDL from earlier fractions using the 1.25 density cut‐off compared to the 1.21 density cut‐off. The cholesterol efflux capacity of early and late eluting fractions from the 1.25 density cut‐off method is increased, suggesting that both smaller and larger higher‐density particles, both with increased efflux capacity, are being isolated with the new method. Gel electrophoresis and Western Blot results from these fractions indicate enrichment of ApoE as well as ApoA‐IV in HDL fractions collected using the 1.25 density cut‐off method, particularly from plasma samples collected in the postprandial vs. the fasted state. In conclusion, we demonstrate that the use of the optimized HDL isolation method yields a broader array of HDL particles, which may be lost when the 1.21 density cut‐off is used.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Microvessel Stenosis and Density Analysis of Post Mortem WMH Detected Using Ultra High Field MRI in Aging, Cerebrovascular and Alzheimer Disease
Authors: Roseborough Austyn , Langdon Kris , Hammond Robert , Pasternak Stephen , Khan Ali , Whitehead Shawn 
Published: June 2021

Cerebral small vessel disease (SVD) affects the microvasculature of the brain, increasing in prevalence with age and associating with cognitive decline and dementia. SVD can be visualized on MR imaging by markers such as hyperintensities within the white matter (WMH). Periventricular WMH within the frontal lobe specifically may have a negative effect on executive functions. Executive dysfunction represents a set of cognitive changes, including processing speed and attention, that can precede memory impairment in Alzheimer disease. The etiology and development of cerebral small vessel disease, including WMH, and it's relation to executive dysfunction is not well understood. Previous work has characterized the presence of stenosis of the arterioles, but little work has focused on the venules. In this study we sought to characterize alterations in microvascular stenosis and density including small and medium venules and arterioles within the frontal white matter. Post mortem 7‐Tesla MR imaging of formalin‐fixed coronal brain sections was performed on 20 brains with a neuropathological diagnosis of normal, cerebrovascular disease or Alzheimer disease. We calculated the percent stenosis within small, medium and large venules and arterioles within both the periventricular and subcortical white matter. Stenosis of the small (<50 um diameter) arterioles and venules within the periventricular white matter specifically was associated with increased severity of periventricular WMH. Small vein and artery stenosis was also associated with the presence of periventricular infarction identified on T1‐weighted imaging. This highlights the utility of multiple sequences when interpreting MRI findings within the white matter. Further study on markers of small vessel disease that affect executive functioning, such as frontal WMH, can help elucidate mechanisms that lead to cognitive decline prior to the onset of memory impairments and dementia.Support or Funding InformationCanada First Research Excellence Fund to BrainsCANCanadian Institutes of Health ResearchCanadian Consortium on Neurodegeneration in AgingThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pericytes Contribute to Retinal Vascular Cell Death in Diabetes: Role of Ephrin‐B2 Signaling
Authors: Coucha Maha M , Barrett Amy C , Abdelsaid Mohammed A 
Published: June 2021

Diabetes‐induced retinal vascular cell death aggravates diabetic retinopathy to proliferative stage and blindness. Pericytes (PCs) share the basement membrane with endothelial cells (EC) in retinal capillaries and play a crucial role in capillaries survival, stability, and angiogenesis. Ephrin‐B2 is a tyrosine kinase that has an essential role in PC/EC communication. The role of Ephrin‐B2 signaling in diabetic retinopathy is under investigation. In the present study, we hypothesized that diabetes alters Ephrin‐B2 signaling in pericytes that increase inflammation and retinal vascular cell death.MethodsType‐II diabetes was induced in male Wistar rats using high‐fat diet and streptozotocin. In vivo, selective reduction of Ephrin‐B2 expression in the retinal PC was achieved using the intraocular injection of AAV‐virus with NG2‐promoter that expresses EphrinB2 shRNA. Retinal trypsin‐digest was used to assess acellular capillary. Human retinal microvascular (HRP) were grown in glucose 25 mM/palmitate 200 uM (HG/Pal) to mimic diabetic conditions. Ephrin‐B2 was silenced in HRPs using siRNA. Cell migration was assessed using a scratch assay. Ephrin‐B2 expression, inflammatory (TNF‐alpha, JNK and IL‐1 beta) and apoptotic (PARP, Caspase‐3) markers were assessed via Western blot and immunohistochemistry.ResultsDiabetes increased Ephrin‐B2 expression in the retinal perivascular areas and pericytes that was confirmed with Western blot analysis. HRP treated with diabetic conditions showed a significant increase in Ephrin‐B2 expression that was associated with increased HPR migration. Silencing Ephrin‐B2 expression significantly reduced PC migration. Increased Ephrin‐B2 expression in pericytes in diabetes was accompanied with an increase in inflammatory markers TNF‐alpha, JNK and IL‐1 beta. In parallel diabetes increases PARP and caspase‐3 cleavage in diabetic retina. Silencing Ephrin‐B2 in retinal PCs decreased retinal vascular inflammation and apoptosis that was in parallel with the decrease in the number of acellular capillaries in retinas of diabetic rats.ConclusionOur results provide novel evidence that diabetes increases Ephrin‐B2 signaling in retinal pericytes that increase retinal vascular inflammation and death. These results identify pericytes‐Ephrin‐B2 signaling as a novel therapeutic target for diabetic retinopathy.Support or Funding InformationAHA, 16SDG30270013 to MA.SUSOP Excellence in Pharmacy Research award to MC.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protective effects of different bile acids against pathological neovascularization
Authors: Thounaojam Menaka C. , Gutsaeva Diana R. , Martin Pamela M. , Bartoli Manuela , Jadeja Ravirajsinh 
Published: June 2021

Emerging evidence suggest that the secondary bile acid ursodeoxycholic acid (UDCA) and its taurine (TUDCA) conjugates, possess neuroprotective and vasculoprotective properties and may exert beneficial effects in a number of neovascular diseases including ischemic retinopathy such as retinopathy of prematurity (ROP). However, the direct activity of UDCA or TUDCA in the treatment of retinal neovascularization has not been previously investigated. Herein, we studied the effects of different bile acids on in vitro angiogenesis in human retinal endothelial cells (HuREC) and retinal vascularization in vivo using the mouse model of oxygen induced retinopathy (OIR), an experimental model of ROP. HuREC were treated with 20 ng/ml of VEGF and/or different doses (10–500 μM) of UDCA or TUDCA for different times. Angiogenic responses were evaluated using tube formation assay. Treatment of HuREC with VEGF significantly induced time dependent tube formation. UDCA dose‐dependently halted VEGF‐induced tube formation, whereas TUDCA had no effect. For the in vivo studies, mice were subjected to OIR and treated with UDCA and TUDCA (administered i.p.) from days P12–P17 (the neovascular stages). Isolectin B4 staining on flat mounted retinal preparations was performed at P17 to evaluate retinal vessel growth and distribution. The results of this analysis revealed that UDCA in the OIR mice halted pathological neovascularization, but did not alter normal vascularization. On the contrary, treatment of the mice with TUDCA failed to block retinal neovascularization, thus confirming the in vitro data. In summary, here we have shown that the bile acids UCDA and TUCDA possess differential ability to modulate VEGF‐mediated angiogenesis effects on HuREC and on retinal pathological neovascularization in vivo. The results of studies support the hypothesis that UDCA could be a potential therapeutic option for ROP as it exerts protection even by systemic administration and does not alter normal progression of vessel formation.Support or Funding InformationThis study is supported by R03HD097660‐01.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Simultaneous knockdown of TLR2 and TLR4 after focal cerebral ischemia facilitates the post‐stroke neurological recovery in rats
Authors: Nalamolu Koteswara Rao , Jorgenson Laura C , Grudzien Natalia A , Choudry Mouneeb M , LaHood Lukas D , Johnson Kendrick D , Klopfenstein Jeffrey D , Pinson David M , Wang David Z , Veeravalli Krishna Kumar 
Published: June 2021

Ischemic stroke is the primary reason for serious long‐term disability. Our recent studies demonstrated that the suppression of toll‐like receptors (TLRs) 2 and 4 alone or in combination attenuated the post‐ischemic inflammation and brain damage. The purpose of this study is to investigate the effect of the simultaneous knockdown of both TLR2 and TLR4 on post‐stroke neurological recovery in young adult male rats. Transient focal cerebral ischemia was induced to male Sprague‐Dawley rats by middle cerebral artery occlusion (MCAO) procedure. A silicone rubber‐coated monofilament suture was introduced into the internal carotid artery via the external and common carotid arteries and pushed until it reaches the origin of the middle cerebral artery followed by removal of the monofilament after 2 hours to initiate reperfusion. Appropriate cohorts of rats were treated with a nanoparticle formulations of TLR2shRNA (T2sh) and TLR4shRNA (T4sh) expressing plasmids (1mg/kg each of T2sh and T4sh) or scrambled sequence inserted vector (SVsh; vehicle) expressing plasmids (2 mg/kg) intravenously via tail vein immediately after reperfusion. The rats from both the cohorts were subjected to various standard neurological tests such as modified neurological severity scores (mNSS), modified adhesive removal (sticky‐tape), beam walking and accelerating rotarod performance before MCAO procedure and at days 1, 3, 5, 7, and 14 after reperfusion. The sensory and motor functions of vehicle treated stroke‐induced rats were markedly impaired when tested at 1d reperfusion followed by a gradual improvement over 14 days. Prevention of the post‐ischemic induction of TLR2 and TLR4 attenuated the degree of neurological impairment and facilitated the magnitude of functional recovery. Therefore, we conclude that the post‐stroke induction of TLR2 and TLR4 in rat ischemic brain contributes to neurological deficits.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aliskiren Effect on Kidney Renin‐Prorenin Stained Cells in a Rat Model for Fat Embolism
Authors: Khalafi Farnaz , Randall Zach , Leupold Brad , Silswal Neerupma , Monaghan Paula , Poisner Alan , Molteni Agostino 
Published: June 2021

IntroductionThe renin angiotensin system (RAS) is involved in the control of blood pressure through its action on the kidney and the treatment by the direct renin inhibitor Aliskiren (Ali) reduces it. We have previously shown in a rat model of fat embolism (FE) that triolein (T) injection leads to a severe inflammatory pulmonary reaction that causes fibrosis in lungs and Ali rescues it. We extended this study to the kidney by evaluating the renin/prorenin (R/P) presence in the kidney with and without Ali in a rat model of fat embolism.MethodsSprague Dawley rats (280–300 grams) were injected with T (0.2 ml IV, n=18) or saline (n=4) through tail vein. The T treated rats were divided into three groups of 6 rats each and one hour later injected with 0.2 ml saline, Ali 50mg/kg or Ali 100mg/kg while 4 controls received saline. After 48 hours, rats were euthanized, the kidneys were dissected out and fixed in 10 % formalin, sectioned and later immunostained for R/P specific antibody (from Abcam). Two pathologists unaware of the slide identity took 10 random images. R/P immunostained cells were quantified by using ImageJ program on medulla and cortical regions where the glomeruli were evident.ResultsIn contrast to our previous results in the lungs of the same animals, the renal sections showed a minimal inflammatory damage without significant vasculitis and fibrosis. There are multiple cells cortical, tubular and glomerular showing the R/P cells immunostaining. It was evident by quantification of these cells by Image J that treatment with saline, T + saline, and T+ Ali resulted in an increase of R/P positive cells versus the saline + saline group but there was no statistically significant difference. However, in the glomeruli of the cortical region, R/P staining was mostly confined in the juxtaglomeruli of saline + saline and T+ saline groups and treatment with both doses of Ali reduced it and was statistically significant.ConclusionAs previously reported for lungs and hearts, R/P positive cells were also present in kidneys. However in kidney, marked differences in several regions were seen after Ali administration. Furthermore, by reducing the R/P evaluation only to glomeruli, the study confirms that Ali pharmacological activity is interacting with the RAS system, and the drug has a potential use for the treatment of the respiratory distress syndrome consequent to the occurrence of fat embolism.Support or Funding InformationSupported by Mary Katherine Geldmacher Research Foundation, St. Louis MO.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The lack of Galectin 3 modifies the evolution of the healing and post‐ myocardial infarct remodeling in mice
Authors: Betazza Maria Celeste , Estevez Florencia Fontana , Cassaglia Pablo , Wilensky Luciana , Cozzi Maria , Truant Sofia Noli , Miksztowicz Veronica , Fernandez Marisa , Naya Nadia Laura Martinez , Volberg Veronica , Berg Gabriela , Casanova Veronica , Morales Celina , Gonzalez German Esteban 
Published: June 2021

Myocardial infarction (MI) is a dynamic process that leads to ventricular remodeling (VR) and heart failure (HF). Previous studies established that Galectin‐3 (Gal‐3), is increased during infarction and it is a prognostic marker of HF. Here, we aimed to study the effects of genetic mutation of Gal3 on macrophages infiltration, fibrosis and MMP‐2 as well as VR after MI in mice. Adult male C57 and Gal3KO mice were subjected to permanent coronary artery ligature and sham. After 1 week (w) post‐MI, echocardiography was performed. LV end‐diastolic diameter (LVEDD) was measured and ejection fraction (EF%) calculated. Then, animals were euthanized. Body (BW) and heart weight (HW), tibial length (TL) and lung wet weight (LW) were measured. Hearts were harvested, snap frozen and storage at −80°C. Infarct size and fibrosis in MI area were quantified in cardiac slices stained with Masson's Trichrome and Picrosirius Red. Myocyte cross sectional area (MCSA) was also measured in slices labeled with wheat germ agglutinin–rhodamine. At 1w post‐MI macrophages (F4/80+ cells) were quantified by flow cytometry and MMP‐2 activity was studied by zymography, both in the MI area.Resultsare expressed as X±SEM; * p<0.05 C57 vs Gal3KO. At 1w post‐MI, the pulmonary congestion assessed by the LW/BW ratio (mg/g) was 9 ± 0.4, 8 ± 0.5 and 9 ± 0.4 in C57 sham, Gal3 KO sham and C57 MI, respectively, but it was severely increased to 15 ± 1.2* in the Gal‐3 KO MI. The macrophage infiltration (%) was reduced from 5.6 ± 0.9 to 2.6 ± 0.4 and the fibrosis (%) in infarct zone was reduced from 30 ± 1.0 to 18 ± 0.4* in C57+MI and Gal3KO+MI respectively. At the same time, MMP2 activity increased from 1 ± 0.1 (C57+MI) to 2 ± 0.4*(Gal3KO+MI). The Infarct size (%) increased from 39 ± 5 to 67± 5* in C57+MI and Gal3KO+MI respectively. Moreover, EF (%) was reduced from 47 ± 2 to 38 ± 3*, while the LVEDD was increased from 4.4 ± 0.1 to 4.8 ± 0.2* in C57+MI and Gal3KO+MI respectively. Fibrosis at remote zone as well as the hypertrophy index evaluated through the HW/BW ratio (mg/g) were similar between groups. In summary, Gal‐3 is essential for the early wound healing and VR in mice. The lack of Gal‐3 reduces the infiltration of macrophages and fibrosis while increasing the MMP‐2 activity, the infarct size and expansion, LV dilation and dysfunction. New studies should focus on the role of Gal3 on the macrophage phenotype profiles, cytokine expression and chronic remodeling.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.