bluuebunny/crossref_metadata_2025_split huggingface dataset:
In‐Vitro Evaluation of the Antidiabetic Effects of Coffee as a Natural Inhibitor of Alpha‐Glucosidase
Authors: Day Xiara , Hill Jenna , Obih John‐Clifford Agbasi , Tolessa Wajira , Obih Patience O 
Published: June 2021

Diabetes mellitus is the seventh leading cause of death in the U.S., and affects over 29 million people. Type 1 diabetes, Type 2 diabetes, and gestational diabetes are the three prominent types. It is a progressive disease, often associated with numerous complications, including, but not limited to, retinopathy, neuropathy, and nephropathy. In 2012, the costs of diabetes exceeded 245 billion dollars. Oral hypoglycemic medications, insulin, and other injectable medications, have been widely used to control Type 2 diabetes, but have undesirable side effects and are expensive. Some agents control postprandial blood glucose levels through inhibition of α‐glucosidase, which delays the cleaving of polysaccharides, preventing the resulting monosaccharides from entering the blood stream. Examples of α‐glucosidase inhibitors are acarbose and miglitol. However, they both have undesirable side effects. Recent studies have pointed toward Phytotherapy, the use of medicinal plants or herbs which have little or no side effects, to treat hyperglycemia. This study investigates the hypoglycemic effects of coffee. Some studies have proposed that coffee may suppress postprandial hyperglycemia. We hypothesized that coffee reduces hyperglycemia by inhibiting alpha‐glucosidase activity. To determine this effect, serial dilutions of 200 mg/mL aqueous coffee extract was mixed with a 0.2 M phosphate buffer and 5 mM p‐Nitrophenyl α‐D‐glucopyranoside in a 96 well plate. The assay quantifying the α‐glucosidase inhibition activity of coffee was performed in‐vitro and measured using a spectrophotometer. Acarbose was used as the positive control of α‐glucosidase inhibition. The IC50 was evaluated, as it is the concentration of the extract required to inhibit 50% of α‐glucosidase activity. From our observation, coffee extract did not inhibit alpha glucosidase at the doses used. More investigations are needed to optimize the approach in assaying the α‐glucosidase inhibition of coffee as well as identifying other mechanisms of action.Support or Funding InformationCenter of Excellence (COE) and BUILD program at Xavier UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Titanium Dioxide TiO2 Induces Loss of Proliferative Capability Related to Ultrastructural Changes in Human Mesenchymal Stem Cells from Wharton Jelly
Authors: Lugo‐Martínez Gabriela , Gutierrez‐Iglesias Gisela Camelia , Parra‐Barrera Alberto , Calzada‐Mendoza Claudia Camelia , Ramos‐Godínez Maria Del Pilar , Cáceres‐Cortés Julio Roberto 
Published: June 2021

IntroductionTitanium dioxide is one of the most widely used mineral pigments in the world in the field of paints, cosmetics, sunscreens, plastics, paper, food, etc. It has been reported that TiO2 at different nanometric scales is considered toxic, depending on the concentration and type of cell that is exposed. TiO2 nanoparticles (TiO2‐NPs) have the ability to cross biological barriers. In vivo models, it cross the blood‐brain barrier and also it could to cross the fetoplacental barrier, although it is unknown if they are capable to internalize in sites called niches of mesenchymal stem cells, which relevant cells sources due to the importance in regeneration of various tissues. The internalization of nanometric TiO2 particles on Wharton's jelly hMSC remains unknown.ObjectiveEvaluate the toxic effect and internalization of TiO2 nanoparticles in hMSCs from Wharton's Jelly.Materials and MethodsWharton gelatin hMSC were cultured and were exposed to three different concentrations (500, 62.5 and 4 μg/mL) of anatase TiO2‐NPs. After 2, 5 and 7 days of exposure, viability tests were performed by alamar blue assay for quantify the toxic effect of TiO2. To identify the location of TiO2‐NPs and morphological changes in the hMSCs, various preparations with different TiO2 concentration were observed by transmission microscopy.ResultsThe results showed that the cellular viability of hMSC decreased significantly at days 2, 5 and 7 of exposure, reduction of viability 40%, 30% and 30% respectively (fig 1). Transmission microscopy showed that the hMSCs had morphological changes and, more specifically, that the TiO2‐NPs took a perinuclear distribution as they accumulated inside the cell, forming a characteristic halo (fig 2). The images showed the internalization of NPs through vesicles, which led to the loss in the continuity of the membrane and the formation of intracytoplasmic spaces representing cellular structural damage in different cellular organelles.ConclusionsThis results are strong evidences that have to be considered in the regulation of the use of nanotechnology in different products containing TiO2 without this affecting public health. Sanitary regulation is suggested in the use of TiO2‐NPs to prevent the health condition as it has been done in different countries.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differentiation of the anti‐coagulant effects of Activated Protein C and Recombinant Thrombomodulin as studied in whole blood and retrieved plasma
Authors: Siddiqui Fakiha , Laddu Rajan , Hoppensteadt Debra , Fareed Jawed 
Published: June 2021

IntroductionActivated protein C (APC) mediates its anti‐coagulant effects by digesting Factor Va & Factor VIIa, whereas recombinant thrombomodulin (rTM) produces its effects by complexing with thrombin and mediating the formation of APC. Both agents have been developed for clinical indications such as Sepsis associated coagulopathy. The purpose of this study is to compare the anti‐coagulant effects of these two agents in deidentified patients whole blood and retrieved plasma.Materials & MethodRecombinant thrombomodulin (recomodulin) was obtained from Asahi kasei Tokyo, Japan. Plasma derived activated protein C was obtained from Hematologic technologies, Burlington Vermont, USA. Both agents were supplemented in 39 individual citrated whole blood collected from patient from clinical coagulation laboratories at a concentration of 2.5ug/ml. Whole blood was analyzed for PT and aPTT on a ST4 instrument (Stago Paris, France). Whole blood was centrifuge at 2500 g to contained plasma samples, which was analyzed for PT & aPTT. Saline supplemented individual blood samples were used as control. Individual blood and plasma results were compiled as group Mean ± SD and Percent change levels were calculated.ResultsIn the whole blood studies, rTM produced a modest increase in the PT assay (2%), in contrast to APC which produced stronger response (36%). In the aPTT assay both agents produced stronger effects, rTM produced a 36% increase whereas APC produced a much stronger increase amounting to 172%. In retrieved plasma rTM produced slight increase in PT assay (2%) while APC produced strong response (20%). In the aPTT assay rTM produced strong effects with 13% however APC produced much stronger effects about 198%. These results were given in Table.ConclusionThese results indicates that APC is a stronger anti‐coagulant then the rTM in both the whole blood and plasma system as measured by the PT and aPTT. Both agents produced stronger anti‐coagulant effects in the aPTT assay suggesting that the intrinsic coagulation pathway is strongly inhibited by these agents in comparison to the extrinsic pathway.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Defining the intracellular redox‐dependent protein signaling networks between cell‐junctions and focal adhesions during intestinal mucosal repair
Authors: Matthews Jason , Reedy April , Jones Rheinallt , Neish Andrew 
Published: June 2021

Mucosal wounds and inflammation of the human intestine are hallmark features of intestinal bowel disease. It is the coordinated intestinal epithelial cell spreading and migration at the wound edge that facilitates mucosal wound closure in a process known as restitution. During restitution, intestinal epithelial cells spread and migrate as a sheet along focal adhesions, or points of basal cellular contact with the extracellular matrix. Cell‐to‐cell contacts of migrating cells are maintained by junction proteins. As restitution begins, epithelial cells produce reactive oxygen species (ROS) by NOX‐1 enzyme. Over time immune cells infiltrate the inflamed and damaged mucosa and increase the oxidative stress by depositing even larger quantities of ROS around and onto the damaged epithelium, leading to cysteine oxidation of a wide variety of cellular and extracellular proteins. As intestinal epithelial cells spread and migrate as a sheet to close the wound, the cellular junctions and the focal adhesions must somehow communicate through a cross‐talk that maintains directional migration and barrier properties. Little is known about the cross‐talk between cell junctions and focal adhesions, or how ROS regulates this mucosal repair program. We have previously reported redox‐dependent subcellular changes of the focal adhesion protein, Cas, localizing to cellular junctions within the epithelium at the edges of mucosal wounds. We hypothesize those redox‐dependent changes in protein signaling drive the mucosal repair program by cysteine‐oxidation and subsequent alteration in kinase/phosphatase activity that produces changes in cell function, and we have sought to elucidate the profile of this protein network. We show here that epithelial cell Cas localization is indeed dependent on oxidation by ROS generated by immune cells during mucosal repair, and provide evidence that similar redox conditions also regulate the cell border accumulation of AKT and FAK. Remarkably, pharmacological inhibition of FAK increased barrier function. Furthermore, we have employed a novel proteomics technique, called Isotope‐Coded Affinity Tag (ICAT) that quantifies the level of proteome‐wide cysteine oxidation in order to further define these signaling modules. Our analysis has revealed redox‐dependent protein signaling driven by MAPK/PAK2/TLN1/actin cytoskeleton interactions, which might regulate Cas, AKT and FAK localization to junctions during mucosal repair. Collectively, these data shed new light on the redox‐dependent junction/focal adhesion cross‐talk during intestinal restitution.Support or Funding InformationCrohn's and Colitis Foundation, Career Development Award # 451678This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Recombinant Fasciola hepatica Fatty Acid Binding Protein Downregulates Inflammatory Cytokines/Chemokines in a Mouse Model of Septic Shock and Shows a Wide Spectrum of Action
Authors: Rosado‐Franco Jose Javier , Benitez‐Ramos Marcos Javier , Ruiz‐Jiménez Caleb , Espino‐Hernández Ana Margarita 
Published: June 2021

Sepsis is the leading cause of death from infections. It involves, both pathogen and host factors and it is characterized by an exacerbation of inflammatory cytokines and chemokines and organ dysfunction and failure. Recently, we reported that a recombinant Fasciola hepatica fatty acid binding protein, termed Fh15, is capable of suppressing the LPS induced expression of inflammatory cytokines such as IL‐1β and TNFα in murine bone marrow derived macrophages in vitro. We also demonstrated that such anti‐inflammatory effect is achieved by targeting specifically the CD14 co‐receptor, which results in a complete suppression of TLR4 activation and makes macrophages irresponsive to subsequent LPS‐stimuli. In the current study, we evaluated the capacity of Fh15 to suppress the TLR4‐activation in vitro, as well as its capacity to therapeutically suppress the cytokine storm in a mouse model of septic shock. We also tested the ability of Fh15 to suppress the activation of multiple TLRs in response to Gram‐positive and Gram‐negative whole bacteria extracts in vitro. For in vivo studies, we used female BALB/c mice that received a lethal intraperitoneal (i.p.) injection of LPS‐E. coli and 1h later received a single i.p. injection with 50μg Fh15. Animals were sacrificed 12h after the LPS challenge. For in vitro experiments, we used THP1‐CD14 cells, which were cultured with Fh15 prior to stimulation with optimized concentrations of 1×108 cells of Klebsiella pneumoniae and Enterococcus faecalis. Results demonstrated that, although Fh15 was unable to stimulate the secretion of any inflammatory cytokine, it significantly suppressed the levels of serum IFNγ (p=0.0425), TNFα (p=0.0008), IL‐1β (p=0.0021) in all animals exposed to lethal doses of LPS. Fh15 also significantly suppressed the levels in serum of two macrophage inflammatory proteins MIP‐1α (p=0.0027) and MIP‐1β (p<0.0001), two chemotactic chemokines that play relevant roles in the inflammatory process. Moreover, we also demonstrated that Fh15 inhibits the stimulation of various TLRs in response to whole bacteria extracts, suggesting that Fh15 could have a broad spectrum of action. These results support the possibility of using Fh15 as an excellent alternative for an anti‐inflammatory drug in pre‐clinical studies in the near future.Support or Funding InformationThis study was supported by MBRS‐RISE R25GM061838‐13, and NIH grants G12MD007600 and 2P40OD012217.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A2A Receptors Are Highly Expressed in Young Patients with Luminal B and Triple Negative Breast Cancer Subtypes and Are Associated with a Higher Proliferate Index
Authors: Guimei Maha , Eladl Mohamed Ahmed , Tawfik Dalia Ra'a Ahmed , Habbal Fuad Wardan , Harrous Abdelkader , El‐Sherbiny Mohamed 
Published: June 2021

Background and ObjectivesNumerous Molecules in the tumor microenvironment orchestrate the body's immune response against tumor cells thereby affecting tumor cell growth and proliferation. Adenosine is produced in the hypoxic tumor milieu and by stimulating its receptors (A2AR), plays an important immunosuppressive role that helps tumor cells evade the body s immune mechanisms. Furthermore, stimulation of these receptors also has a profound role in promoting the tumor vasculature and contributing to tumor aggressiveness. The present study aimed at investigating A2AR expression in breast cancer tissue and its association with different clinicopathological parameters of the tumor.MethodsThis study was conducted on 30 formalin‐fixed paraffin‐embedded (FFPE) human breast cancer tissues. Sections were immunohistochemically stained with of A2AR antibodies. Clinical and pathological data were retrospectively obtained from the patients' records in the archives of the pathology department at Alexandria University, Egypt.ResultsA2ARs were expressed not only on the tumor‐infiltrating immune cells (TILs) but were also found on the tumor cells themselves in 73.3% (n=22) of breast cancer tumors. Their expression on the tumor cells was significantly higher in younger age patients (< 50 years) (88.2%) compared to older ones (54.5%) [p=0.044]. A2AR expression was also significantly associated with a Luminal B as well as triple‐negative tumors [p=0.028] compared to Luminal A and Her2‐enriched tumors. Patients expressing A2A receptors had a significantly higher expression of Progesterone receptors [P=0.031] compared to ER expression [P=0.098]. The majority of A2AR expressing tumors had a mitotic index score of 2 [p=0.013] as well as a significantly higher proliferative index (ki‐67 >20%) [p=0.018]. No association was observed between A2AR expression and tumor size, type, grade, Lymph node status, the percentage of TILs or patient survival after 2 years of follow‐up.ConclusionOur study confirmed that A2AR were strongly expressed on breast cancer tumor cells in younger patients particularly those of luminal B and triple negative subtypes. Their expression was associated with a significantly higher proliferative index suggesting a possible role of these receptors in imparting a more aggressive phenotype and that targeting these receptors could have a potential role in improving the outcome of these patients.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In vitro effect of 5G8‐specific antibodies on Giardia lamblia trophozoites
Authors: Rascon Raul , Dominguez Victor , Garzon Thania , Samaniego Brenda , Valdez Alejandra , Quintero Jael , Rascon Lucila , Velazquez Carlos 
Published: June 2021

Giardiasis is one of the most common gastrointestinal diseases worldwide, caused by the protozoan parasite Giardia lamblia. Although several immunogenic proteins of the parasite have been identified, the capacity of inducing a protective humoral response has been scarcely evaluated. Our research group have identified and characterized the immunogenic 5G8 protein, a 70 kDa Variant‐specific surface protein (VSP). The administration of 5G8‐specific monoclonal antibodies (AcMo 5G8.B5) to infected animal decreased the parasite intestinal load. The aim of the present study was to evaluate the in vitro effect of 5G8 specific antibodies on G. lamblia trophozoites. We isolated 5G8 protein by passive elution from SDS‐PAGE (membrane protein enriched fraction of trophozoites lysates). C3H/HeJ mice were intraperitoneally immunized with isolated 5G8 protein to generate polyclonal antibodies. GS/M‐83‐H7 5G8 (+) trophozoites were incubated with monoclonal and polyclonal antibodies specific to the 5G8 protein, and their agglutination capacity was evaluated. After 24, 48 and 72 hours incubation, cell proliferation was determinated and the expression of 5G8 protein on trophozoites was evaluated by Flow Cytometry. Polyclonal antibodies recognized 97 % of GS/M‐83‐H7 5G8 (+) trophozoite population, and a single band of 70 kDa (SDS‐PAGE). The AcMo 5G8.B5 and polyclonal antibodies had the ability to agglutinate G. lamblia trophozoites in vitro after 60 min of antibody exposure. Antibody treatments inhibited the in vitro G. lamblia growth. Isotype control antibody and preimmunized serum showed no inhibition of trophozoites proliferation. Surface 5G8 protein expression did not change during antibody treatments. Altogether, our data suggest a possible in vitro cytostatic effect of 5G8 specific antibodies on G. lamblia trophozoites. These findings put forward the capacity of the 5G8 protein to induce a protective humoral response against the infection.Support or Funding InformationCONACYT: CB‐2010‐01/155224This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of Myocardial Ca2+ Dynamics by the G Protein‐Coupled Estrogen Receptor
Authors: Whitcomb Victoria , Wauson Eric , Matnishian Vahe , Tran Quang‐Kim 
Published: June 2021

The G protein‐coupled estrogen receptor (GPER) participates in many cardioprotective activities. However, the mechanisms of such activities are not known. We have begun to investigate the effects of alterations in GPER activity on myocardial signaling via the beta‐1 adrenergic receptor. In freshly isolated primary murine cardiomyocytes, β1AR agonist isoproterenol triggers robust Ca2+ signals that are oscillatory and dependent on extracellular entry. GPER activation dose‐dependently inhibits the isoproterenol‐induced Ca2+ signals. On the other hand, GPER antagonism exerts the opposite effects. Similar effects were observed in H9C2 cardiomyocytes. Mechanisms of these observations were examined on various components of myocardial Ca2+ signaling. The data indicate that GPER activity is involved in β1AR‐mediated Ca2+ signaling in the myocardium.Support or Funding InformationAHA grant 15SDG25090279 (to E Wauson) and NIH Grant HL112184 (to Q‐K Tran)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synaptic mechanisms underlying the rapid‐acting antidepressant actions of ketamine
Authors: Zobel Allison , Pope Brock , Rehman Zarin , Herring Laura , Wauson Eric , Graves Lee , Duric Vanja , Yuan LiLian 
Published: June 2021

Ketamine at sub‐anesthetic doses has shown promising results as a potent and fast‐acting antidepressant. Evidence from animal model studies suggests that the functional restoration by ketamine is associated with activation of MAPK and mTOR signaling cascades in the prefrontal cortex (PFC) within a few hours of ketamine administration, followed by a second wave of synaptic protein upregulation. Together, these molecular events lead to rapid synaptogenesis and reversal of neural atrophy. Furthermore, (2R, 6R)‐hydroxynorketamine (HNK) has recently been identified as the main active component of ketamine metabolism. HNK is believed to be responsible for the antidepressant actions of ketamine, but with minimal side effects, and potentially through NMDAR‐independent mechanism of action. However, critical gaps exist in understanding the exact molecular mechanisms of ketamine actions. Thus, it would be of great importance and interest to delineate the molecular mechanisms that translate mTOR activation into synaptogenesis‐based structural remodeling. Cofilin and its upstream pathways, ideally positioned to bridge the gap between mTOR activation and spine formation, have emerged from an unbiased kinome/phosphoproteome screening for targets of ketamine treatment. As the major cytoskeletal component of dendritic spines, actin controls spine morphology and formation through changes in its polymerization state. Actin polymerization is negatively regulated by activation of cofilin protein. Phospho‐antibody based assays and direct or indirect measurement of kinase activity are currently underway to validate those targets in acutely and chronically stressed animal models. We anticipate addressing the significance of cofilin phosphorylation and actin polymerization in antidepressant responses of ketamine and HNK, and whether those responses are modulated by other factors including stress and sex.Support or Funding InformationNIH grant MH108043 and IOER fundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.