bluuebunny/crossref_metadata_2025_split huggingface dataset:
Long Term Effectiveness of A Dietary Portfolio of Cholesterol‐Lowering Foods in Hypercholesterolemic Subjects
Authors: Kendall Cyril WC , Nguyen Tri H , Fozdar Nishant , Faulkner Dorothea A , Marchie Augustine , Wong Julia MW , Josse Andrea R , Srichaikul Kristie K , Vidgen Edward , Holmes Candice , Josse Robert G , Leiter Lawrence A , Singer William , Jenkins David JA 
Published: June 2021

Objective: To determine the long term effectiveness of advice to consume a dietary portfolio of cholesterol‐lowering foods on risk factors for heart disease.Methods: Sixty‐six hyperlipidemic subjects were prescribed diets high in plant sterols (1.0 g/1000 kcal), soy protein (22.5 g/1000 kcal), viscous fibers (10 g/1000 kcal) and almonds (23 g/1000 kcal). Thirty subjects completed 3 years.Results: LDL‐cholesterol (LDL‐C) was significantly reduced at years 1, 2, and 3 with mean reductions of 17.7%, 11.2%, and 10.1%, respectively (P<0.01, n=30). HDL‐cholesterol (HDL‐C) was significantly raised at years 1, 2, and 3 with mean increases of 3.9%, 3.1% and 4.5%, respectively (P<0.01, n=30). Changes in LDL‐C significantly correlated with compliance to the dietary portfolio. Subjects with greater than 75% compliance to the dietary portfolio achieved a mean 24% reduction in LDL‐C.Conclusions: A sustained long term benefit of the dietary portfolio was seen in hyperlipidemic subjects with lower LDL‐C and raised HDL‐C over a 3‐year period. The effectiveness of the dietary portfolio was greater with >20% LDL‐C reductions seen in subjects having over 75% compliance.Funding: Canada Research Chair Endowment of the Federal Government of Canada; the Canadian Natural Sciences and Engineering Research Council of Canada; Loblaw Brands Limited; the Almond Board of California; Unilever Canada and Unilever Research & Development, Vlaardingen, The Netherlands; and Yves Veggie Cuisine.
Liver mesenchymal Thy‐1 cells: important player in oval cell mediated liver regeneration
Authors: Yovchev Mladen I , Zhang Jialin , Neufeld David , Grozdanov Petar , Dabeva Mariana D 
Published: June 2021

Recently, we and others have reported that Thy1 antigen is expressed in rat liver myofibroblasts. To study further the properties and the function of these cells, we purified Thy‐1+ cells by MACS and FACS and analyzed their gene expression pattern using Affymetrix Rat expression array 230 2.0, RT‐PCR and IF microscopy. Thy‐1+ cells are highly activated in models of oval cell‐mediated liver regeneration, tightly surrounding the oval cell pseudoducts. Thy‐1+ cells appear, proliferate and disappear from the liver in parallel with oval cells. Thy‐1+ cells express mRNA for genes considered specific for stellate cells: desmin, v‐cam, gfap, pdgfrβ, integrin β3; for myofibroblasts: αsma, elastin, fibulin‐2, fibrilin‐1; for matrix remodeling proteins: mmp2, collagen I‐a1, ‐a2, III and IV; for mesenchymal stromal cells: CD54, CD73, CD105, CD166, SDF‐1; and for growth factors and cytokines that can stimulate oval cell growth: Il‐6, hgf, tweak. Co‐expression of Thy‐1 antigen with desmin, αSma and Pdgfrβ was confirmed by double IF microscopy. RT‐PCR analysis of isolated individual Thy‐1 expressing clones confirmed the heterogeneity of these cells. Conclusion: liver mesenchymal Thy‐1+ cells are the major supporting cells in the oval cells niche.Supported by NIH grant R01 DK59321 (to M.D.D.)
High glucose alters tubular epithelial cell expression of molecules in the renin‐angiotensin axis
Authors: Kruss Szilvia , Danziger Robert , Kotlo Kumar , Setty Suman 
Published: June 2021

Purpose: Aminopeptidase N (APN), a type II membrane ectoenzyme also known as CD13, is a major component of Glut‐4 containing vesicles. It has been postulated to regulate secretory function in general and glucose transport in particular. The present study was performed to determine the effect of glucose and insulin on the expression and activity of APN in renal epithelial cells.Methods: A heightened glucose milieu was created by culturing immortalized HK‐2 cells in 25 mM glucose as compared to the normal 5 mM glucose and a control for hyperosmolality (cells grown in 5 mM glucose with 20 mM mannitol)(OC). The model was validated by studying the altered growth and adhesion characteristics of the HK‐2 cells in the three conditions. Assays for APN activity and APN transcript abundance were performed on cells cultured in the three conditions. The effect of insulin and lisinopril on glucose‐induced changes APN activity was examined.Results: APN transcript, protein, and activity were greater in cells treated with 25 mM vs 5 mM glucose and the same in cells treated with OC vs 5 mM glucose. Insulin reduced APN activity to that observed in low glucose (5mM) levels (p<0.05).Conclusions: Glucose is capable of stimulating APN in renal epithelial cells. This stimulation was blocked by insulin, raising the possibilities that APN plays a role in glucose homeostasis in general and in the pathogenesis of diabetes specifically.
Knock down of AQP1 in HMEC‐1 cells and WM115 cells changes the organisation of the cytoskeleton
Authors: Porta caterina AM La , Monzani Elena , Facchetti Floriana , perego Carla 
Published: June 2021

Aquaporin 1 (AQP1) is specifically and strongly expressed in most microvasculature endothelial cells outside of the brain. Herein we have investigated the role of AQP1 in both microvascular endothelial (HMEC‐1) and in a human melanoma cell line (WM115). The knock down of AQP1 by RNA interference modified in both cells the organization of F‐actin as well as reduced the migration and invasive capacity significantly. In order to better understand the intracellular mechanisms elicited by AQP‐1, we have investigated the role of Lin‐7, a PDZ protein which is involved in the recruitment of receptors, adhesion molecules and signalling proteins and that can interact with cytoskeleton through beta‐catenin.Our results showed that AQP1 co‐immunoprecipitated with Lin‐7 and the knock down of AQP‐1 decreased the level of expression of Lin‐7 and beta‐catenin. Interestingly, the inhibition of proteasome contrasted partially such a decrease.All together, our findings show, for the first time, that AQP1 triggers an intracellular mechanisms modulating and stabilizing the organization of cytoskeleton through Lin‐7/beta‐catenin interaction. A model for these functions of AQP1, at least for endothelial and melanoma cells, is presented.
Synthesis and biological evaluation of soluble epoxide hydrolase (sEH) inhibitors: t‐AUCB and its derivatives
Authors: HWANG SUNG HEE , Tsai Hsing‐Ju , Morisseau Christophe , Liu Jun‐Yan , Hammock Bruce D. 
Published: June 2021

Soluble epoxide hydrolase (sEH, EC ) is involved in the metabolism of endogenously derived fatty acid epoxides, such as arachidonic acid, linoleic acid, and other lipid epoxides. Epoxyeicosatrienoic acids (EETs), the cytochrome P450 epoxygenase products of arachidonic acid, are known modulators of blood pressure and inflammation. A series of N,N′‐disubstituted ureas having a conformationally restricted cis‐ or trans‐1,4‐cyclohexane alpha to the urea were prepared and tested as sEH inhibitors. This series of compounds showed low nanomolar to picomolar activities against recombinant human sEH. Both isomers showed similar potencies, but the trans‐isomers were more metabolically stable in human hepatic microsomes. Furthermore, these new potent inhibitors show a greater metabolic stability in vivo than previously described sEH inhibitors. Among them, trans‐4‐[4‐(3‐Adamantan‐1‐yl‐ureido)‐cyclohexyloxy]‐benzoic acid (t‐AUCB, IC50 = 1.3 ± 0.05 nM) had excellent oral bioavailability (98%, n = 3) and blood area under the curve (AUC) in dogs and was effective in vivo to treat hypotension in lipopolysaccharide (LPS) challenged murine models. The synthesis, SAR, and PK properties of this series as well as the scale‐up process and efficacy of t‐AUCB along with PK profile of corresponding esters and salts of t‐AUCB also will be presented.(supported in part by NIEHS grant ES02710, NIEHS Superfund grant P42 ES04699, and NIEHS Center grant P30 ES05707)
Microsomal epoxide hydrolase E404D mutation influences the turnover of epoxyeicosatrienoic acids
Authors: Burgener Julia , Marowsky Anne , Cronin Annette , Arand Michael 
Published: June 2021

Traditionally, the microsomal epoxide hydrolase is thought of as a detoxifying enzyme as it hydrolyzes many carcinogenic epoxides to their corresponding dihydrodiols. Recent data by our group that substantiate earlier findings implicate that the mEH may also have a physiological role other than detoxification. The enzyme is capable of hydrolyzing epoxyeicosatrienoic acids (EETs), a class of signaling molecules known to regulate a wide variety of physiological functions, such as blood pressure, angiogenesis, inflammation and apoptosis. Thus, by metabolizing the EETs to the corresponding dihydrodiols, the mEH might also be implicated in the modulation of such processes.The enzymatic reaction mechanism involves a nucleophilic attack followed by a hydrolysis via a water‐activating charge relay (H431 and E404). An artificial E404D mutant shows a faster metabolization rate compared to the wild type enzyme. This more efficient variant has, so far, not been found in the animal kingdom. We hypothesize that this may be connected to an altered EET metabolism by the E404D variant. So far, we have expressed recombinant wild type and E404D mEHs from rat, mouse and human and presently analyze their turnover rate with EETs using LC‐MS/MS.We expect that our findings will elucidate some more details about the physiological role of mEH and may help to explain why in animals the less active version of mEH is established.
Soluble epoxide hydrolase inhibition attenuates the development of renovascular hypertension in 2K1C rats.
Authors: Huskova Zuzana , Kopkan Libor , Walkowska Agnieszka , Vanourkova Zdenka , Hammock Bruce D. , Imig John D. , Cervenka Ludek 
Published: June 2021

The present study tested the hypothesis that inhibition of soluble epoxide hydrolase (sEH), that increases the level of epoxyeicosatrienoic acids, would attenuate the development of renovascular hypertension in 2K1C rats. Renovascular hypertension was induced by placing a silver clip on the left renal artery in Wistar rats for 25 days. The sEH inhibitor, c‐AUCB was given in drinking water (0.5 mg/day) starting 48 hours before the renal artery clipping. Blood pressure (BP) was monitored by radiotelemetry in conscious animals during 30 days of experimental period. At the end of the experiment the ratio of left heart ventricular weight (mg) to tibial length (mm) (LVW/TL) was used for assessing cardiac hypertrophy.After 30 days of experimental period, systolic BP was not different in sham operated rats treated with sEH inhibitor (106±2 to 104±3 mmHg; n=5) compared to sham control group (107±3 to 112±4 mmHg; n=4). However, the sEH inhibitor, c‐AUCB, attenuated the development of hypertension in treated 2K1C rats (110±2 to 138±3 mmHg, p<0.05; n=5) compared to untreated 2K1C (113±4 to 161±7 mmHg, p<0.05; n=6). Moreover, significantly higher LVW/TL ratio in untreated 2K1C compared to sham control group (26±1 vs. 20±1) was partially attenuated in 2K1C rats treated with sEH inhibitor (23±1, p<0.05).These data demonstrate that sEH inhibition attenuates the development of renovascular hypertension.
Self‐organization and pattern formation in calcifying vascular stem cells
Authors: Demer Linda L. , Yochelis Arik , Tintut Yin , Garfinkel Alan 
Published: June 2021

We previously observed patterns of evenly spaced stripes of calcified cellular aggregates in cultures of calcifying vascular cells (CVC), bovine smooth muscle cells purified by dilutional cloning. We hypothesized that the patterns were formed by a reaction‐diffusion process driven by interaction of a known morphogen, bone morphogenetic protein‐2 (BMP‐2), with its inhibitor, matrix GLA protein (MGP). We used mathematical models based on the biochemical and physical properties of BMP‐2 (autocatalytic, chemoattractant, and osteogenic) and MGP (inhibitory of BMP‐2 and rapidly diffusing).Computer simulations based on the math models predicted 2 experimental findings. Over 10–14 days, CVC cultures treated with exogenous MGP produced evenly spaced spots (nodules) rather than stripes, and CVC treated with warfarin (an inhibitor of MGP) produced stripes at double the density of untreated cultures. In addition, the models predicted conversion of diffuse cell density to focal nodules. In vivo, when CVC were implanted for 8 wks in subcutaneous diffusion chambers in hyperlipidemic mice, they produced tissue nodules resembling calcified atherosclerotic plaque.Thus, calcified atherosclerotic lesions may arise from cellular aggregates formed by reaction‐diffusion processes. Harnessing this mechanism may ultimately allow clinical control of focal vs. diffuse vascular disease.
Role of epigenetic histone modifications during early embryonic stem cell differentiation
Authors: Fritsch Michael Kevin , Murdoch Fern , Singer Don , Lee Elliot , McCool Kevin , Alexanian Ruben , Oduro Akua 
Published: June 2021

Early embryonic stem cell differentiation includes “exit” from the pluripotent state followed by “commitment” to a specific lineage. During both of these processes there are genome‐wide changes in gene transcription levels that are regulated, at least in part, by epigenetic changes in histone tail modifications. We have demonstrated both global and gene specific changes in histone tail modifications. We focused on the role of these marks as they regulate the earliest stages of differentiation in mouse ES cells.Histone H3 phosphoacetylation (activating mark) is increased within the first 3 days of differentiation. Blocking general phosphorylation pathways markedly slowed exit from the pluripotent state. Blocking MSK1, the specific kinase responsible for phosphorylation on histone H3 still resulted in exit, but with diminished expression of early mesoderm markers. We also demonstrated a gene specific increase in acetylation (activating) of histones H3 and H4 and a global increase in acetylation of H4 as cells began lineage commitment. A pulse of the histone deacetylase inhibitor, TSA, maintained expression of markers of the undifferentiated state while also inducing expression of early mesoderm markers. TSA effects on later lineage commitment are being studied.We propose that epigenetic modifications play an important role in early cell fate decisions.This work was supported by NIH RO1 DK064243 from NIDDK.
Connecting development and disease pathways through zebrafish chemical biology
Authors: Peterson Randall 
Published: June 2021

The molecular pathways that direct normal embryonic development often resurface as causative factors in adult diseases. For example, bone morphogenetic proteins (BMPs) establish the embryonic dorsal‐ventral axis, but disruption of BMP signaling contributes to the anemia of chronic disease. Similarly, AML1 is a master regulator of embryonic hematopoiesis, but AML1 mutations are causative in a significant proportion of cases of acute myeloid leukemia (AML). This intimate connection between development and disease offers hope that experimentally tractable developmental models may provide new avenues to the study and treatment of adult cancers.We have developed methodologies that enable high‐throughput small molecule screens to be conducted with whole, living zebrafish. These screens allow us to discover small molecules that modify developmental pathways and also exhibit high specificity, good pharmacological properties, and low toxicity. The small molecules discovered are useful probes for studying embryonic development and can often be developed further to study and treat the cancer associated with the targeted developmental pathway. We are currently exploring the developmental pathways involved in myocardial infarction recovery, AML, and anemia and testing whether our small molecule modifiers will also be helpful for studying or treating these diseases.
Stem/Progenitor Cell Mobilization for Tissue Regeneration
Authors: Lee Sang Jin , Van Dyke Mark , Atala Anthony , Yoo James 
Published: June 2021

IntroductionIn this study we explored the possibility of utilizing host stem or progenitor cells for tissue regeneration in vivo. We investigated whether these cells could be mobilized into target specific sites for regenerative medicine applications.Materials and MethodsPolyglycolide polymers were implanted subcutaneously in mice to recruit host cells. The scaffolds were retrieved at 1, 2, 3 and 4 weeks after implantation for analyses. Cells contained within the retrieved scaffolds were cultured and differentiated into osteogenic, endothelial, myogenic, and adipogenic cell lineages using the respective cell differentiation conditions.Results and DiscussionThe retrieved implants showed a progressive increase in neovascularization and host cell infiltration with time. Tissue infiltrates within the scaffolds showed the evidence of collagen deposition after 2 weeks of implantation and continued to increase over time. The culture of infiltrated cells showed the expression of stem cell markers and these cells were able to differentiate into various cell lineages.ConclusionsThis study demonstrates that stem or progenitor cells can be mobilized and recruited into target specific sites. These findings suggest that host stem cells may be maneuvered to achieve tissue regeneration in vivo using a target specific scaffold system.
Proliferation and extracellular matrix protein expression in vascular smooth muscle cells cultured for aortic valve tissue engineering
Authors: Appleton Andrew James Edward , Appleton Christopher Thomas George , Boughner Derek , Rogers Kem 
Published: June 2021

OBJECTIVES: Vascular smooth muscle cells are a potential autologous cell source for aortic valve tissue engineering. We hypothesized that combining basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and platelet derived growth factor (PDGF) with transforming growth factor beta‐1 (TGF‐β1) treatment would allow temporal control of rat aortic smooth muscle cell (RASMC) proliferation and ECM production.METHODS: Growth factor (alone or in combination with TGF‐β1) ability to induce myofibrolast‐like phenotype in RASMCs in monolayer culture was assessed. Cell proliferation was measured by MTT assay following treatment with growth factor combinations. Total RNA was isolated to assess ECM gene expression by real‐time PCR.RESULTS: Combinations of growth factors which included PDGF showed the greatest increases in proliferation. Immunofluorescence for alpha‐smooth muscle actin (α‐SMA) in conditioned cultures demonstrated an inverse correlation between proliferation and myofibroblast phenotype, with the combination of TGF‐β1+bFGF+EGF showing the greatest α‐SMA expression. Finally, TGF‐β1+EGF+PDGF treatment showed a significant increase in versican, fibronectin and type I collagen mRNA expression.CONCLUSION: A combination of TGF‐β1+EGF+PDGF, can increase RASMC proliferation and induce ECM gene expression profiles similar to those of native aortic valve interstitial cells.
Increased expression of SOX2 in Ammon's Horn Sclerosis: Further evidence for neurogenesis in epilepsy
Authors: Eschbacher Jennifer Mary , Coons Stephen 
Published: June 2021

Neurogenesis persists throughout adult human life in the dentate gyrus and subventricular zone. Recent evidence points to increased neurogenesis in the hippocampus of epileptic animal models. The abnormal migration of new neurons to ectopic locations in the dentate gyrus, in conjunction with aberrant integration of new neurons into the granule cell layer, has been hypothesized to contribute to increased seizure activity. Many studies have also shown an increase in neurogenesis secondary to CNS insults such as stroke and trauma. We therefore hypothesized that we would see increased expression of SOX2, a neural stem cell marker, in hippocampal sclerosis. Using immunohistochemical analysis, we examined the hippocampus from nine patients for SOX2 expression. Seven were from patients with a known history of temporal lobe epilepsy and histologically confirmed Ammon's horn sclerosis and two were from normal autopsy controls. We found many immunoreactive cells throughout the hippocampous, including the subventricular zone, in all seven seizure cases, and no expression of sox2 in the autopsy controls. These results provide further evidence in support of active neurogenesis in epilepsy and suggest that SOX2 plays a role in neurogenesis in hippocampal sclerosis.This research was funded and supported by Barrow Neurological Institute.
Characterization and Differential Inhibition of Topoisomerase IB from the Nosomial Multidrug Resistant Pathogen Pseudomonas aeruginosa by Fluoroquinolone Drugs
Authors: Jain Teesta , Grove Anne 
Published: June 2021

P. aeruginosa is an opportunistic pathogen responsible for many life‐threatening conditions in immunocompromised patients of neutropenic cancer and bone marrow transplant. P. aeruginosa encodes a putative topoisomerase (PAT) that shares sequence and biochemical characteristics with the vaccinia topoisomerase IB (VT). PAT has been purified and shown to relax supercoiled DNA without ATP but in the presence of MgCl2/MnCl2. The predicted tyrosine at the active site (Tyr292) when mutated to phenylalanine does not perform the relaxation reaction. In addition, the catalytic domain with the truncated amino terminal topo (81–333) performs the same repertoire of reactions as the wild type enzyme.Fluoroquinolones are a subset of quinolones that are a group of antibacterials. The quinolones inhibit the type II topoisomerase exemplified by DNA gyrase. Sparfloxacin is a synthetic fluoroquinolone, which belongs to the same class as ofloxacin and norfloxacin. Although the PAT is not inhibited by norfloxacin, lomefloxacin, enrofloxacin, moxifloxacin and camptothecin, it is inhibited by sparfloxacin at higher concentrations. Like VT, PAT is resistant to camptothecin and the specific fluoroquinolones which inhibit the viral enzyme.
An interaction between Mex67p and Upf1p at the terminal step of mRNA export facilitates mRNA translation
Authors: Agosto Abby Joyce , Cajigas Iván , Guzzardo Paloma , Krebber Heike , González Carlos 
Published: June 2021

Gene expression relies on the communication between the processes of transcription, mRNA processing, export and translation. Once synthesized in the nucleus, an mRNA must become competent for its translocation across the nuclear pore. Formation of an mRNA particle (mRNP) suitable for export is accomplished by the binding of soluble export factors to the nascent mRNA. Following transport, mRNPs must undergo structural rearrangements in the cytoplasm in order to be translated. We have gathered information to support this mRNP remodeling step. Specifically, we have found that Mex67p interacts with Upf1p. We provide evidence to support the notion that the Mex67p‐Upf1p interaction is neither required for the activation of the NMD pathway or for poly‐(A)+ mRNA export. Instead, Upf1p and Mex67p are critical to facilitate the conversion from an export‐competent mRNP to a translation‐competent mRNP, since cells lacking both proteins demonstrate severe defects in protein synthesis. Specifically, our data demonstrate that these cells are unable to efficiently initiate mRNA translation and recognize translation termination codons. We propose that in the cytoplasm, Upf1p communicates the Mex67p‐bound mRNAs with the protein synthesis machinery to facilitate translational competency and couple the terminal step of mRNA export to mRNA translation.Supported by RISE Award 2R25GM61151, NIH U54 Award CA96297‐03, NIH‐NIGMS SCoRE Award GM008102‐305, and P20 RR 016174 from NCRR‐NIH.