bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co‐localization of Hepatocellular Uptake Transporters in Detergent Resistant Membranes (Lipid Rafts)
Authors: Steiert Brianna , Idowu Jessica Y. , Dougherty Larissa , Hagenbuch Bruno 
Published: June 2021

BackgroundWe have previously shown that several transporters located in the basolateral membrane of human hepatocytes (OCT1, NTCP, OATP1B1, and OATP1B3) interact with each other. These interactions affect both surface expression and function. Currently, transporter studies are performed using cell lines that express a single transporter at a time without considering how transporter interactions will affect function. Determining how transporters co‐localize, e.g. in detergent resistant lipid rafts or via direct protein‐protein interactions, has far‐reaching implications for drug metabolism and pharmacological studies.AimThe purpose of this study is to test the hypothesis that these transporters co‐localize with well‐known lipid raft marker proteins.MethodsFollowing homogenization in 1% Triton X‐100 on ice, lipid rafts were isolated from HEK293 cells expressing NTCP, OCT1, OATP1B1, and OATP1B3, or from frozen human liver samples. Homogenates in 40% sucrose were overlaid with a discontinuous 5% to 35% sucrose gradient. After centrifugation for 18 hours at 197,000 × gav, gradients were fractionated, and aliquots were analyzed using western blotting. The blots were probed with antibodies against positive (caveolin and Na+/K+ ATPase) and negative (clathrin) lipid raft markers, as well as with transporter‐specific antibodies.ResultsIn human liver, caveolin, Na+/K+ ATPase, OATP1B1, OATP1B3, and OCT1 were detected in similar fractions. OCT1 also was found in clathrin‐positive fractions towards the bottom of the gradient. In HEK293 cells, NTCP and OCT1 co‐localized with caveolin in the middle of the gradient and again, OCT1 was also found towards the bottom of the gradient.ConclusionThe detection of NTCP, OCT1, OATP1B1, and OATP1B3 along with the positive lipid raft markers caveolin and Na+/K+ ATPase indicates that co‐localization of these transporters occurs, at least in part, because of their residence in detergent resistant membrane domains. Future experiments are aimed at identifying the functional consequences of this co‐localization.Support or Funding InformationP20 GM103549, P30 GM118247, and GM077336This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Caveolin‐1 in Normal and Oviduct Pregnant Tissue
Authors: Wang Lin , Wang Qianhua 
Published: June 2021

Objective To detect the expression of caveolin‐1 in normal and tubal pregnant tissue of human and explore its clinic significance.Methods 20 fresh clinical operative tubal tissue specimens were collected from the women who were diagnosed oviduct pregnancy. 20 fresh normal oviduct tissues were collected from the women who were diagnosed uterine benign tumors and they were divided into two groups: the proliferative phase and secretory phase according to the morphological characters. The expression of caveolin‐1 was detected by immunohistochemical method and analyzed by image analysis technique.Results The positive expression of caveolin‐1 in the normal human oviduct tissue of secretory phase was stronger than that of proliferative phase. And the positive expression of caveolin‐1 in tubal pregnant tissue was lower than that in the normal human oviduct tissue of secretory phase. The differences were significant (P<0.01). The positive expression of caveolin‐1 can be detected in cytotrophoblast cell but not in syncytotrophoblast cell of villus.Conclusion The lower expression of caveolin‐1 in the oviduct pregnant tissue may play an important role in the oviduct pregnancy.
The non‐coding RNA gadd7 is a regulator of lipotoxic‐induced ROS and ER stress
Authors: Thomas Rita , Listenberger Laura L , Harp Jeff D , Ory Daniel S , Schaffer Jean E 
Published: June 2021

To elucidate molecular events in the lipotoxicity pathway, we used retroviral promoter trap mutagenesis to generate a mutant Chinese hamster ovary (CHO) cell line resistant to palmitate‐induced cell death. In this mutant, insertion of the provirus has disrupted one allele of the non‐coding RNA (ncRNA) growth arrest DNA‐damage inducible gene 7 (gadd7), creating a model of haploinsufficiency. Knockdown of gadd7 in wild type (wt) cells also causes palmitate‐resistance. Because reactive oxygen species (ROS) induces gadd7 and palmitate generates ROS, we hypothesized that palmitate induces gadd7 expression in a ROS‐dependent manner. Palmitate treatment of wt cells induces gadd7 expression, but this is diminished by co‐treatment with an antioxidant. Although gadd7 haploinsufficient cells generate ROS in response to palmitate, induction is decreased compared to wt cells. These data suggest a feed forward loop between gadd7 and ROS generation. Since ROS generation contributes to palmitate‐induced ER stress, we assessed gadd7's role in palmitate‐induced ER stress. Induction of ER stress related genes is significantly reduced in palmitate treated gadd7 haploinsufficient and knockdown cells, indicating that gadd7 expression is necessary for palmitate‐induced ER stress. Together these data implicate a ncRNA in propagation of oxidative stress and in regulation of ER stress responses.NIH F31 (RT) and DK064989 (JES)
microRNA influence on host response to Francisella infection
Authors: Cremer Thomas John , Hunter Melissa G. , Elton Terry S. , Marsh Clay B. , Tridandapani Susheela , Butchar Jonathan P. 
Published: June 2021

The gram‐negative bacterium Francisella tularensis has the ability to subvert host immune response and is the causative agent of the disease tularemia. Previous work has identified the roles of PI3K, Akt and SHIP in modulating host resistance to Francisella. However, the mechanisms of F. tularensis host cell subversion are still poorly understood.microRNAs (miRs) have emerged as an important regulatory system in the modulation of host innate immune response. Since miRs have the potential to regulate key immune functions such as NFkappaB activity and TLR signaling, we examined whether they played a role during Francisella infection. We performed a miR array analysis of human myeloid cells infected with Francisella. This led to the identification of several miRs that were significantly up‐ or down‐regulated by Francisella. We subsequently verified many of these changes using real‐time RT‐PCR in both human and murine cells. Currently we are examining the roles of these miRs by identifying their targets and by modulating miR activity itself during Francisella infection. Interestingly, our data so far indicate that SHIP and PI3K/Akt regulate the expression of several miRs. As specific miR targets are identified within the context of Francisella infection, it is likely that novel therapeutic targets will be found, and that the mechanisms of Francisella host cell subversion will be further elucidated.
Influence of Metal Ions on Immune Reactivity
Authors: Chan Erwin PH , Zellweger Rene , Filgueira Luis 
Published: June 2021

Orthopaedic metal implants are routinely used in bone fracture repair and for joint replacement therapies. However, approximately 15% of recipients suffer from inflammatory reactions and aseptic loosening. One possible cause is an immune reactivity towards the metal implants. Metal implants in patients undergo biocorrosion releasing metal ions and wear particles. Metal ions are able to circulate in the body and bind to extracellular and/or intracellular proteins. Any immune reactivity by patients would be initiated by professional antigen presenting cells, dendritic cells (DCs) and effected by CD4+ T‐cell.For this in vitro study, Titanium (IV) and Vanadium (IV) ions were chosen as they are major components in current metal implants. Using energy filtered transmission electron microscopy, uptake of Titanium (IV) by human DCs showed intracellular accumulation as phosphorus containing titanium complexes at the surface and intracellular membranes, as well as in ribosomal and nuclear chromatin structures. Changes in the DC surface markers after exposure to metal ions measured using flow cytometry. Increase in expression of co‐stimulatory molecules (CD80, CD86 and CD40) was seen in vanadium exposed DCs. Upregulation of cytokines IL4, IL10, IFN‐g and TNF‐a was detected in the supernatants of DCs exposed to titanium (IV). Finally, the ability of metal exposed DCs to interact with lymphocytes was also investigated through flow cytometry. Decrease in T‐lymphocytic CD25 was seen after 5 day culture with Titanium (IV) treated DCs whilst vanadium treated DCs were able to maintain CD25 expression.The reactivity towards various metal ions varies with individuals. It is suggested that responders towards metals would have a reduced regulatory capability through CD4+CD25+ cells, allowing the proliferation of inflammatory osteo‐immune Th17cells.
ICAM‐1 expression is required for regulatory T cell localization in the lungs of M tuberculosis infected mice
Authors: Windish Hillarie Plessner , Flynn JoAnne L 
Published: June 2021

Mycobacterium tuberculosis (Mtb) is a widespread human pathogen that persists in the lungs of latently infected individuals. Formation of the granuloma, a hallmark lesion of tuberculosis, allows leukocytes to control infection with Mtb. We hypothesized that the adhesion molecule ICAM‐1 plays a role in the granuloma formation and in the cell to cell communication that takes place at the locus of infection While previous studies in ICAM‐1−/− mice have indicated that ICAM‐1 is required for long‐term control over Mtb infection, the mechanism remains unclear. In this study, ICAM‐1−/− mice infected with Mtb survived similarly to wild type controls with equivalent bacterial burden. Although granuloma formation appeared normal in the absence of ICAM‐1, higher overall lymphocytic infiltrate was observed by H&E staining and flow cytometry. Higher T cell numbers was apparently due to inhibited contraction in the absence of ICAM‐1 following primary Mtb infection. T regulatory cells were also significantly reduced in ICAM‐1−/− mice, which could play a role on T cell contraction. In this study we have both investigated the role of T regulatory cells in controlling T lymphocytes. Results suggest that ICAM‐1 expression may help to control lung pathology by supporting the function of T regulatory cells in the lungs. Future studies will attempt to elucidate the mechanism by which ICAM‐1 affects T regulatory cell recruitment to or retention in the lungs.Funding provided by NIH R01 HL71241
Interferon (IFN) beta ‐1a induces IL‐27 and IL‐12, and down‐regulates IL‐23 and IL‐1beta in dendritic cells (DCs), establishing inhibitory conditions for Th‐17 cell differentiation: Implications for treatment of multiple sclerosis (MS)
Authors: Ramgolam Vinod , Speer Danielle , Markovic‐Plese Silva 
Published: June 2021

Background: Th‐17 cells have been implicated in the pathogenesis of MS. DC‐derived cytokines IL‐6, IL‐1 beta and IL‐23 promote, whereas IL‐27 and IL‐12 suppress Th‐17 cells differentiation. Objective: To investigate the effect of IFN beta‐1a, a standard therapy for MS, on DC‐derived cytokines and signaling events that regulate Th‐17 cells differentiation. Materials/Method: Monocyte‐derived mature DCs from 13 donors were cultured in the absence or presence of IFN beta‐1a (1000 IU/ml) and analyzed for cytokine gene expression by quantitative RT‐PCR and for signal transduction by Western blots. Results: IFN beta‐1a increased the expression of IL‐27 (191.5‐fold, p=0.002), IL‐12p35 (6.7‐fold, p=0.008) and decreased IL‐23 (−3.7 fold, p=0.002), and IL‐1 beta (−2.7‐fold, p=0.004) in DCs derived from 10 healthy donors. The results were confirmed in 3 MS patients. IFN beta‐1a induced phosphorylation of STAT‐1, a transcription factor for IL‐27. The role of STAT1 as a transcription factor for IL‐27 was confirmed in the experiments where STAT‐1 gene silencing reduced IL‐27 transcription by 30% in IFN beta‐1a‐treated DCs. IFN beta‐1a induced the expression of SOCS3, which may mediate inhibition of IL‐23 and IL‐1b expression. Conclusion: IFN beta‐1 induces IL‐27 expression via STAT‐1 activation and inhibits IL‐23 and IL‐1beta, possibly through the induction of SOCS3, promoting the suppression of Th‐17 cell differentiation in MS.The study was supported by EMD Serono.
Using gamma‐cytokine complexes to improve antigen specific CD8 T cell responses in tumor‐bearing mice
Authors: Verdeil Gregory , Marquardt Kristi L , Biggs Judith A , Surh Charles D , Sherman Linda A 
Published: June 2021

In the absence of inflammatory signals, the exposure of T cells to tumor antigen in the periphery results in tolerance. Remaining tumor specific T cells are usually of low avidity or anergic. Immunotherapy aims at reversing this tolerance to induce an efficient T cell response. STAT5 Cytokines of the interleukin‐2 family signal through STAT5 and are candidates to increase T cell anti‐tumor response. The effects of these cytokines are greatly increased when the cytokine is complexed with a specific antibody.We have studied the potential of cytokine complexes to serve as adjuvants for CD8 cells activation in a murine model, RIP‐Tag2‐HA, in which pancreatic insulomas expressing the influenza hemagglutinin (HA) as a surrogate antigen develop spontaneously. HA specific CD8 cells become effector after 3 days of treatment with IL‐2 complexes and antigen in vivo. This differentiation leads to the elimination of endogenous tumors by high avidity HA specific CD8 cells. Survival of CD8 cells is increased by using the TLR ligands polyIC or by extending the length of treatment with IL‐2 complexes. Under such conditions, even low avidity HA specific CD8 cells eliminate the tumors. We further show that cytokine complexes also increase survival of adoptively transferred, ex‐vivo activated HA specific CD8 cells. These results indicate that IL‐2 complexes are strong adjuvants that greatly enhance tumor vaccines or immunotherapy.
Sperm protein 17 is expressed in primary and metastatic breast cancer
Authors: Chiriva‐Internati Maurizio , Ferrari Raffaele , Yu Yuefei , D’Cunha Nicholas , Grizzi Fabio , Cobos Everardo , Jenkins Marjorie R 
Published: June 2021

Objective. Breast carcinoma (BC) is a major health concern for millions of women. The cancer testis antigen (CTA) Sperm protein 17 (Sp17), has been identified in multiple myeloma and ovarian cancer. This study evaluated Sp17 expression in both normal breast tissue and primary invasive ductal BC.Methods. We analyzed Sp17 expression in 7 normal breast tissues, 22 primary BCs by immunohistochemistry (IHC) and three human cell by Western Blotting (WB).Summary of Results. No immunoreactivity was recognized in normal breast tissues while BCs showed strong cytoplasmic immunoreactivity with 15/22 positive cases (70%) for Sp17 protein.Conclusions. The data demonstrate the expression of Sp17 in breast cancer. Further studies will elucidate the role of Sp17 in BC, and its potential use as a BC immunotherapeutic target antigen.
Precision and validity of self‐reported anthropometry in Guatemalan adolescents
Authors: Campos J Raquel , Aquino Brenda , Solomons Noel W. 
Published: June 2021

Background:The validity self‐reported height (Ht) and weight (Wt) in adolescence has often been reported, but rarely validated. Its reproducibility is unknown.Objectives:To determine the precision and validity of self‐reported Ht and Wt in adolescents from 11 to 14 y of age.Methods:Nutritional questionnaires, including self‐reported Ht and Wt, were performed in low‐income urban Guatemalan schools at 2‐wk intervals in 100 subjects each, in 4 age‐groups (11,12,13,14 y), equally divided by sexes (F,M). Direct Ht and Wt measurements were made after the 2nd interview. BMIs were derived as kg/m2. Similarities in group‐medians and strength of individual associations were calculated.Results:Not all participants self‐reported a Ht and a Wt value on both or either occasion; 30% reported both values both times. For 118 subjects reporting the Ht twice, Spearman correlation was r=0.90 and the medians differed by 2.5 cm F and 0.5 cm M. For Wt repetition in the same subjects, r=0.93 and the medians differed by 1.1 lbs F and 0.0 lbs M. With respect to true BMI and the derived value from self‐reported on the same day, r=0.65.Conclusion:Self‐reported anthropometry may provide valuable estimates at the group level, but is not adequate for individual assessment in this age group.
Consumption of pistachios significantly reduces IL‐1 expression in isolated human mononuclear cells
Authors: Gebauer Sarah K , Zhang Jun , Heuvel John P Vanden , Kris‐Etherton Penny M 
Published: June 2021

The effects of nuts on CVD risk have been well studied, but little is known about pistachios. A randomized, crossover, controlled feeding study was conducted to compare the effects of 2 doses of pistachios on CVD risk factors in hypercholesterolemic adults (n=22; LDL‐C = 133 ± 4 mg/dl). Subjects were fed 3 isoenergetic diets for 4 wk each after a 2‐wk run‐in diet [35% total fat (TF), 11% SFA, 8% PUFA, 13% MUFA]. Diets included a Step‐I diet without pistachios (25% TF, 8% SFA, 5% PUFA, 9% MUFA), with 1.5 oz/d (1.5 PD, 10% en; 30% TF, 8% SFA, 6% PUFA, 12% MUFA), and 3 oz/d (3 PD, 20% en; 34% TF, 8% SFA, 8% PUFA, 15% MUFA). There was a significant (P < .05) reduction in IL‐1 expression after the 1.5 PD (−36%) and 3 PD (−44%). Those with low serum CRP (<1 mg/L) had a greater decrease in IL‐1 expression (P<.05). After consumption of the 3 PD, IL‐1 expression was directly associated with expression of TNFα (r = .9, P < .0001), CRP (r = .5, P < .05), IL‐6 (r = .7, P < .001) and ICAM (r = .8, P < .0001). Changes in IL‐1 expression were directly associated with plasma PUFA (r = .3, P < .05), specifically 18:3 n‐6 (r = .4, P < .01) and 20:4 n‐6 (r = .4, P < .01), and inversely associated with MUFA (r = −.5, P < 0.001), specifically 18:1 (r = −.6, P < 0.0001). Including pistachios in a heart healthy diet reduces IL‐1 expression, thus beneficially affecting CVD inflammatory risk status.Funded by CA Pistachio Commission and partially by PSU GCRC (NIH grant M01RR10732)
Augmented Myocyte Protection And Proliferation In The Infarcted Heart Following Treatment With Cell Lysate From Pharmacologically Preconditioned Mesenchymal Stem Cells
Authors: Afzal Muhammad Rizwan , Haider Husnain Kh , Idris Niagara Muhammad , Jiang Shujia , Rahman Shahid , Ashraf Muhammad 
Published: June 2021

We previously showed that preconditioning of mesenchymal stem cells (MSCs) with diazoxide upregulates expression of multiple trophic factors. We hypothesize that intramyocardial injection of cell lysate from these MSCs (pcMSCs) will prevent myocyte apoptosis, stimulate proliferation and attenuate infarct size. Real time PCR showed higher expression of a series of trophic factors in pcMSCs. Cardiomyocytes pretreated with pcMSC lysate were more resistant to oxidant stress and showed higher phosphorylation of PKC å and PKC ä compared to cardiomyocytes treated with lysate from non preconditioned cells. This effect was blocked by Chelerythrin. For in vivo studies, 50 μl of pcMSC lysate (group 1) or non‐pcMSC lysate (Group 2) or DMEM (group 3) was injected into the acutely infarcted wall. Echocardiography showed improved LV function in group 1 at 6 weeks. Hearts were harvested at day 7 and 6 weeks for Masson Trichrome staining that showed significantly attenuated infarct size, reduced area of fibrosis and well preserved LV wall thickness in group 1 compared to other groups. Immunostaing for Ki67 and á‐actinin showed higher number of proliferating cardiomyocytes in group 1. Capillary density was also highest in the peri‐infarct region in group 1.Conclusion: The trophic factors rich lysate from pcMSC triggers proliferation of cardiomyocytes and preserves LV wall thickness and function.
Novel free radical scavengers protect retinal cells and decrease elevated intraocular pressure in a rat ocular hypertension model
Authors: Hosseini Alireza , Lattanzio Frank A. , Schellenberg Karl A. , Samudre Sandeep S , Shaeffer James , Williams Patricia B. 
Published: June 2021

Glaucoma is a neurodegenerative disease with damage retinal cells (RC) often associated with elevated intraocular pressure (IOP). Novel free radical scavengers (FRS) with esterified ion chelator and reducing side groups on a methoxypolyethylene glycol backbone (MPs) were evaluated as anti–glaucoma agents capable of both IOP reduction and RC protection.The operated eye of surgical rat glaucoma model received topical MPs or amifostine, a clinical FRS (all agents 20 ml, 0.3 – 100 mM). IOP was measured via Goldmann tonometer. Slit lamp, confocal microscopy and chronic toxicity studies were performed on normal rats (10 or 87 mM MPs, 3x/day for 30 days). 14C MPs were used for pharmacokinetic studies. Intravitreal injection with NMDA (2μL, 10–40mM) to naïve rats induces RC ERG and histological losses in 2 weeks. This damage was challenged with co–administered intravitreal MPs (n=5–6).Topical MPs (3–100 mM), but not amifostine, decreased IOP up to 29.6 % for T1/2s of 1–2 hrs. MPs also caused no acute or chronic ocular/systemic toxicity. 14C topical MPs showed ocular penetration with elimination via urine. MPs also reduced NMDA induced RC loss and A and B waves decreases. In vitro isolated rat and bovine RC assays are currently evaluating the role of free radical generation, elevated intracellular calcium and mitochondrial depolarization in the MPs mechanism of action in vivo.Supported in part by CHRB and AHAF.
Mathematical Model of Tissue Oxygenation in Early Sepsis
Authors: Fraser Graham Mathew , Goldman Daniel , Ellis Christopher G. 
Published: June 2021

The defining characteristic of sepsis is a progressive microvascular dysfunction remote to the locus of infection. Our objective was to quantify the time course of changing tissue PO2 due to microvascular injury in early sepsis using an established mathematical model. Experimental data obtained from a rat cecal‐ligation and perforation (CLP) model of sepsis has demonstrated a progressive FCD loss at 2, 3 and 4 hours (11±4%, 15±1%, 21±5% SHAM vs 18±3%, 32±5%, 48±7% CLP respectively). Microvascular blood flow was recorded in skeletal muscle using dual wavelength intra‐vital video microscopy. Capillaries were analyzed at baseline in vivo for O2 saturations and hemodynamics. Custom vascular mapping software was used to reconstruct the real geometry (length, diameter, tissue depth and connections between vessels) of the experimental network (Volume 420 × 420 × 60 μm). A constant consumption computational model applied to the FCD conditions at each time point found mean tissue PO2 was not different between SHAM and CLP at 2 hours (34.2±4.3 vs 33.3±4.2 mmHg) or 3 hours (33.0±4.3 vs 31.2±4.5 mmHg) post. At 4 hours tissue PO2 was significantly different between SHAM and CLP (32.7±4.3 vs 25.9±6.5 mmHg, p < 0.01). This simulation suggests that a substantial FCD loss precedes a significant decrease in tissue oxygenation due to diffusional exchange between adjacent capillaries.Supported by CIHR grant to DG and CGE.