bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inductively Coupled Plasma Optical Emission Spectrometry (ICP‐OES) Analysis of Lead Bioaccumulation in Chlamydomonas reinhardtii and the Role of Cia7 Gene
Authors: Vazquez Cano Erick U. , Reyes Victoria , Ynalvez Ruby A. 
Published: June 2021

The indiscriminate use of heavy metals in human activities has become a threat for the biochemical equilibrium and geochemical cycles of nature. To survive in heavy‐metal polluted environments, some microorganisms have developed a variety of detoxifying mechanisms. An excellent model organism for the study of heavy metal tolerance and homeostasis is Chlamydomonas reinhardtii, a unicellular green haploid microalga. The purpose of this research project was to identify the function of the novel gene Cia7 in C. reinhardtii, and specifically, to determine if it has a role in heavy metal tolerance. It was hypothesized that wild type C. reinhardtii (CC4425) would exhibit less bioaccumulation and higher resistance heavy metal stress; while a mutant strain in which Cia7 has been suppressed (CC5013), would exhibit inhibited growth and higher metal bioaccumulation compared to the wild type. In this study, both strains of C. reinhardtii were treated with either 0 μM or 150 μM of Pb(NO3)2 solution. Lead bioaccumulation after three days of treatment was determined by inductively coupled plasma optical emission spectrometry (ICP‐OES). Chlorophyll concentration and cell growth analysis were also performed. Both strains of C. reinhardtii were treated with increasing concentrations of lead, and absorbance readings at 663, 645, 730 nm were taken on 3‐day intervals over a 15‐day treatment period. Results from ICP‐OES analysis showed no difference in lead bioaccumulation between both strains. The wild type strain (CC4425) demonstrated a relatively higher growth rate than the mutant (CC5013). Chlorophyll concentration and growth in the wild type strain decrease as the concentration of lead in solution increases. On the other hand, chlorophyll concentration and growth in the mutant strain was observed to increase with a higher treatment of lead.Support or Funding InformationACT on Ideas ‐ Texas A&M International University/College of Arts and Sciences ‐ Texas A&M International UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the Role, Expression and Interactions of FAP‐1 in S. cerevisiae, Cultivated in a Nitrogen‐limited Media
Authors: Velez Astrid Carolina Rodriguez , Matos Elsie Pares 
Published: June 2021

The protein FAP‐1 was found in the cytoplasm of Saccharomyces cerevisiae, and it confers resistance to the immunosuppressive drug and antifungal Rapamycin, by competing with the drug for the binding site, which is mutually exclusive for both, thus preventing the formation of the Fpr1p‐Rapamycin complex. This complex inhibits the products of the TORC1 which regulates essential pathways that control some cytoplasmic and nuclear events, and some nutrient regulated genes. In preliminary studies, it has been observed that FAP1 needs to be overexpressed in other to confer this resistance to the yeast. The complex FAP1:Fpr1p has been also observed in the absent of Rapamycin, especially when some nutrient levels are low. By using these observations, we proposed that the concentration of FAP1 might be inversely proportional to the availability of nutrients in culturing media. To prove this hypothesis, the strain CFY7‐PJ69‐6A of Saccharomyces cerevisiae will be cultured in Nitrogen‐limited media and at different time periods. The technique qPCR was used to quantify the amount of fap1 RNA available at different concentrations of ammonium sulfate (the nitrogen source), and at incubation points of 3 hrs, 6 hrs, 9 hrs and 12 hrs. We found that the expression of fap1 is inversely proportional to the availability of nitrogen in culturing media, whereas the growth and division was directly proportional to the nitrogen availability. The expression levels observed with the qPCR could be useful to optimize bioprocesses at industry by monitoring FAP1 in order to determine the availability of target nutrients in media in yeast cultures. Currently, we are interested in the identification of protein‐DNA interactions via Chromatin Immunoprecipitation, to determine which genes this transcription factor might regulate.Support or Funding InformationThis work is supported by the NIH‐MARC‐U‐STAR Grants: 5T34GM008419‐25 and 2T34GM008419‐26, and Puerto Rico's Louis Stoke Alliance for Minority Participation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Distinct Roles of Dietary Fat and Sugar in the Development of Obesity, Insulin Resistance, Atherosclerosis and Cardiac Dysfunction in LDL Receptor Knockout Mice
Authors: Perazza Laís Rossi , Daniel Noemie , Dubois Marie Julie , Pilon Geneviève , Mitchelle Patricia , Le Quang Khai , Lachance Dominic , Plante Eric , Varin Thibaut , Bouchareb Rihab , Mathieu Patrick , Pouliot Yves , Gauthier Sylvie , Roy Denis , Asselin Claude , Blais Mylène , Lessard Martin , Marette André 
Published: June 2021

The increased consumption of high caloric foods has been associated with the obesity epidemic and a higher incidence of cardiometabolic disease, but the specific role of dietary fat and sugar remains unclear. There is growing evidence that high intake of added sugar contributes to cardiovascular disease (CVD) but whether this is independent from high fat content in the diet remains to be established. To this end, our objective was to determine the impact of high sucrose intake on the features of metabolic syndrome and CVD in a mouse model of atherosclerosis.We used atherosclerosis prone LDLr−/− ApoB100/100 (LRKOB100) mouse fed either a low‐fat/high‐sucrose (LFHS) or a high‐fat/low‐sucrose (HFLS) diet supplemented with 0.2% cholesterol for 24 weeks.Body weight gain and whole‐body fat mass were greater in HFLS‐fed vs LFHS‐fed mice. HFLS feeding promoted higher liver triglyceride (TG) deposition and plasma hypercholesterolemia when compared to LFHS feeding. HFLS‐fed animals were more insulin resistant than LFHS as revealed by a higher HOMA‐IR index. Unexpectedly, while LFHS‐fed mice were less prone to metabolic impairments, they had significant atherosclerosis development as revealed by extensive aortic plaque formation in comparison to HFLS‐fed mice. Consistent with atherosclerotic plaque formation, echocardiography further suggested that LFHS‐fed LRKOB100 mice developed left ventricle eccentric hypertrophy (LVEH) after only 12 weeks of feeding as compared to HFLS‐fed mice. Determination of the inflammatory status by multiplex analysis revealed that HFLS‐fed mice had elevated hepatic IFN‐γ, circulating MCP‐1 and PAI‐1. Conversely, LFHS‐related atherosclerosis was linked with increased hepatic levels of IL‐1β, IL‐6, RANTES and TNF‐α. In addition to hepatic inflammation, LFHS fed animals showed higher amount of TNF‐α and IFN‐γ in visceral adipose tissue, consistent with activation of metabolic inflammation.Our results indicate that dietary sugar and fat have distinct roles in the development of cardiometabolic diseases in LRKOB100 mice. While high fat intake promotes obesity and metabolic alterations, high sugar intake promotes atherosclerosis and cardiac dysfunction, with a major effect on the hepatic inflammatory profile. Further investigations are needed to unravel the underlying mechanisms by which high sugar intake promotes the atherosclerotic process.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antioxidant effect of Justicia spicigera in liver from diabetic rats
Authors: Murillo‐Villicaña Marina , Martínez‐Mora José A. , Noriega‐Cisneros Ruth , Manzo‐Avalos Salvador , Salgado‐Garciglia Rafael , Cortés‐Rojo Christian , Montoya‐Pérez Rocío , Saavedra‐Molina Alfredo 
Published: June 2021

The oxidative stress is a factor that contributes to the development of complications in the diabetes disease; however, its effects can be counteracted by the use of exogenous antioxidants, which are found in some plants. That is why people look to the traditional medicine in search of the therapeutic treatment. Justicia spicigera has been demonstrated to have the capacity of reduce glycemia levels; nevertheless the effects in non‐insulin dependent‐organs like liver, has not been reported. During administration of the ethanol extract for 30 days, the blood glucose and weight data were taken every 5 days. Once the treatment concludes, the rats were sacrificed. The corporal weight, blood glucose, cholesterol‐Very Low Density Lipoprotein (VLDL), triglycerides, total lipids and liver profile; were diminished in the diabetic condition and normalized with the application of ethanol extract of J. Spicigera (EJS). Meanwhile catalase activity shown a significant increase in control diabetic rats and a decrease in their activity with the application of the extract, without any effect in normoglucemic rats. There is also a tendency to increase the GSH/GSSG ratio and a decrease in total glutathione levels in diabetic rats. In conclusion, the ethanol extract of Justicia spicigera is considered to reduce oxidative stress in liver in STZ‐induced diabetes rats.Support or Funding InformationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the elongated cell phenotype of Escherichia coli overexpressing the lysophospholipase PldB
Authors: Georgiou George S. , Garrett Teresa A. 
Published: June 2021

The overexpression of the enzyme PldB leads to increased levels of the headgroup‐acylated glycerophospholipid, acyl phosphatidylglycerol (acyl PG), in the membranes of Escherichia coli. The role of acyl PG in E. coli has yet to be fully determined. Interestingly, PldB overexpression leads to an elongated phenotype, suggesting a role for acyl PG in cell length and division. To gain a better understanding of this phenotype, wildtype MG1655 and cells lacking the chromosomal pldB (ΔpldB) were transformed with an empty arabinose‐inducible plasmid (pBAD), and a plasmid containing pldB (pPldB). These four strains (MG1655/pBAD, MG1655/pPldB, ΔpldB/pBAD, and ΔpldB/pPldB) were grown to mid‐log phase and imaged via scanning electron microscopy. Quantification of cell lengths from SEM images showed a significant difference in the average length of the wild‐type strain, MG1655/pBAD (1.85 μm ± 0.46 μm), versus MG1655/pPldB (2.69 μm ± 1.12 μm) and ΔpldB/pPldB (3.02 μm ± 2.42 μm). Furthermore, many of the elongated cells appeared to contain no cleavage furrows suggesting that they failed to complete cellular division. Visualization of the chromosomal DNA by staining with DAPI revealed that PldB overexpressing strains contained multiple unsegregated nucleoids within individual cells, providing more evidence that PldB overexpression leads to irregularities in cellular division. Future research will focus on separation of the inner and outer membranes of E. coli overexpressing acyl‐PG in order to assess the lipid compositions of each respective membrane and elucidate where acyl‐PG migrates and integrates once formed. Additionally, the contractile Z‐ring of E. coli overexpressing acyl‐PG will be visualized via fluorescence microscopy to further investigate the relationship between acyl‐PG and cellular division.Support or Funding InformationSupported by National Science Foundation Research at Undergraduate Institutions grant #1516805.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulating the activity of Akt by inhibiting the pleckstrin homology domain‐PtdIns(3,4,5)P3 interaction using flavonoids
Authors: Kang Yerin , Lee Woonwoo , Yoon Youngdae 
Published: June 2021

Since over‐activation of cellular signalling was frequently observed from a wide range of cancer types, the down‐regulation of signalling is regarded as a goal for drug development. A serine‐threonine kinase, AKT, playing a pivotal role in tumour progression was also over‐activated in cancer cells; thereby AKT was believed as a critical therapeutic target for cancer intervention. In this study, we targeted the pleckstrin homology (PH) domain to down‐regulate AKT activity via inhibiting phosphatidylinositol‐3,4,5‐triphosphate (PIP3) binding using by small natural molecules, flavonoids. The inhibition rates of flavonoids were evaluated by liposome pull‐down assay coupled with fluorescence spectrophotometry, and several flavonoids including 3,6‐dihydroxyflavone and 6,2′‐dihydroxyflavone showed about 40 and 50 % of inhibition at 10μM. Moreover, it was confirmed that flavonoids targeting PH domain‐PIP3 interaction decreased the activity of AKT in cellular levels. And then, we further analyzed the relationship between structure and activity using QSAR to find optimum structural moiety because of the structural similarity of flavonoids. Although it needed more study to derive optimum moiety of flavonoids as anticancer drugs for targeting AKT, we believed that this finding would provide a strong evidence to regulate AKT activity by targeting the regulatory domain instead of catalytic kinase domain.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of the Senescent Preadipocyte Secretome on Skeletal Muscle Cells
Authors: White Thomas A. , Mazula Daniel , Schafer Marissa , Giorgadze Nino , Aversa Zaira , Kirkland James L. , LeBrasseur Nathan K. 
Published: June 2021

Adipocytes accumulate within and around skeletal muscle with advancing age. Adipose progenitor cells, preadipocytes, are prone to cellular senescence, a state of stable growth arrest associated with the secretion of cytokines, chemokines and matrix remodeling proteins, collectively referred to as the senescence‐associated secretory phenotype (SASP). Considerable evidence implicates the SASP in age‐related tissue inflammation and degeneration; however, its impact on skeletal muscle is poorly understood. Herein, we examined the effects of conditioned media (CM) collected from healthy human (control CM) and senescent by irradiation (senescent CM) preadipocytes on human skeletal muscle cells (SkMCs). Compared to control CM, senescent CM had more than 10‐fold higher concentrations of IL‐6, IL‐1β, IL‐8, PAI‐1, MCP‐1 and TNFα as quantified by a multiplexed antibody‐based assay. Senescent CM increased expression of IL‐6 (5‐fold), IL‐1α (1.6‐fold), IL‐1β (3.5‐fold), MCP‐1 (2.5‐fold), PAI‐1 (1.7‐fold) and MMP‐3 (3‐fold) in differentiated SkMCs relative to control CM as measured by qPCR (all p < 0.05). Senescent CM increased proliferation of undifferentiated SKMCs by 20%; however, inhibited differentiation relative to control CM per the fusion index. Preliminary data further suggest senescent CM compromises insulin‐ and IGF‐1‐mediated activation of Akt. Collectively, our data suggest that the SASP of senescent preadipocytes may drive inflammation in SkMCs and compromise their differentiation and, potentially, response to metabolic an anabolic stimuli. These effects may contribute to age‐related changes in skeletal muscle health and regeneration.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Engineering riboswitch‐based whole cell biosensors in Escherichia coli
Authors: Zhao Maryann , Page Katharine , Liu Jane 
Published: June 2021

In vitro diagnostics (IVD) have become increasingly popular tool for tackling global health concerns, but conventional IVD tests, such as microscopy and nucleic‐acid amplification methods, are often expensive and complex to use, limiting their use in low‐resource settings. Whole‐cell biosensors in bacterial systems offer an attractive solution as an easy‐to‐use, portable device that non‐specialists could use for clinical purposes outside clinical locations. However, a strategy is needed to engineer ligand specific sensors for new targets. An appealing candidate for a sensing module is the riboswitch. Genetic RNA switches, termed riboswitches, are genetic regulatory elements found in the 5′‐UTR (untranslated region) of some prokaryotic mRNA and contain two functional components: an aptamer binding domain and an expression platform that modulates downstream gene expression. We developed an engineering platform coupling genetic selections and Fluorescence‐Activated Cell Sorting (FACS) screens to identify novel riboswitches from a 108 random‐sequence library in which we replaced the aptamer sequence of the ThiM#2 riboswitch with a degenerate 40‐nucleotide sequence. In a proof‐of‐principle validation of the platform, we identified novel riboswitches for the small molecule theophylline, which are distinct from the existing synthetic theophylline switches. Our best riboswitch hit displays a 2.3‐fold activation in response to theophylline and does not respond to the structurally unrelated molecule thiamine. We performed directed evolution on this riboswitch hit by mutagenizing its sequence and performing additional rounds of selections and screens, resulting in a variant with a 3.6‐fold activation. Current efforts are focused on assessing the efficacy of the selection platform when isolating riboswitches for dopamine. Furthermore, a variant of the engineering platform is being used side‐by‐side to compare if a dopamine riboswitch can be selected from a library composed of an in vitro selected aptamer coupled with a 15‐nucleotide degenerate sequence. We anticipate that these efforts will provide general guidelines toward the development of novel riboswitches for a variety of natural and non‐natural ligands.Support or Funding InformationNSF CBET 1258307Arnold and Mabel Beckman FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Vasopressin infusion throughout pregnancy causes placental pathology in mice consistent with preeclampsia
Authors: Gibson‐Corley Katherine N. , Sandgren Jeremy A. , Perschbacher Katherine J. , Zhang Shao Y. , Deng Guorui , Santillan Donna A. , Santillan Mark K. , Grobe Justin L. 
Published: June 2021

Human preeclampsia (PE) is associated with elevated secretion of arginine vasopressin (AVP), and chronic infusion of AVP into pregnant mice is sufficient to model PE by causing hypertension, renal glomerular endotheliosis, proteinuria, fetal placental hypoxia, and growth restriction. Early stages of PE are associated with defective trophoblast invasion of maternal spiral arteries, leading to decreased artery diameter and placental oxygenation. AVP infusion (24 ng/hr, sc) reduced placental growth factor mRNA and protein at gestational day (GD) 17.5. Therefore, we performed histopathologic analyses of placentas collected from mice at GD12.5 infused with saline or AVP, with the hypothesis that AVP leads to early placental PE phenotypes. This preliminary cohort demonstrated an increased urine protein content (saline n=7 27.1±2.7, AVP n=11 44.1±2.5 mg/mL; p<0.01), trends toward elevated mid‐gestational systolic blood pressure (104±2.1, 110±2.2 mmHg; p=0.09), and similar changes in heart rate (Δ from baseline: 37.1±15.5, 38.8±11.8 bpm; p=0.93). GD12.5 placentas were then routinely processed and stained with HE or cytokeratin‐8 to examine morphological changes induced by AVP. AVP infusion had no significant effects junctional zone (saline n=11 383±18, AVP n=13 376±15 μm; p=0.76) or decidua layer (652±21, 585±33 μm; p=0.12) thicknesses but significantly reduced labyrinth layer thickness (727±26, 640±13 μm; p<0.05). AVP caused a reduction in average maximum spiral artery diameter within the decidua (161±12, 125±7 μm; p<0.05) and reduced total spiral artery number (8.1±0.8, 4.8±0.4, p<0.01), but no difference in the maximum invasion depth of CK8‐positive trophoblasts (348±22, 302±23 μm; p=0.16). We conclude that AVP infusion is sufficient to induce key mid‐gestational features of PE in pregnant mice, including reduced spiral artery diameter. Such morphological changes may be associated with the reduced placental growth factor expression in this model.Support or Funding InformationNational Institutes of Health (GM067795, HL134850, HL084207), the American Heart Association (13SDG143400012, 15SFRN23480000, 15SFRN23730000, 15SFRN23760002, 15SFRN23860007, 16PRE30980043, 15UFEL25850040, 14PRE20380401), the American Physiological Society (UGREF & UGSRF programs), University of Iowa Center for Hypertension Research, and the Roy J. Carver Trust.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chemorepulsion as a novel therapeutic concept to inhibit pancreatic cancer metastasis
Authors: Niclou Benoit , Li Xiaoran , Zessler Astrid , Adam Rosalyn , Briscoe David , Bielenberg Diane 
Published: June 2021

Pancreatic ductal adenocarcinoma (PDAC) has a high mortality rate primarily due to the absence of effective pharmacologic treatments and the limited effects of surgical resection in the early stages of the disease. The most frequent cause of death from the disease is due to its tendency to metastasize to the liver, at which point the chances of long‐term survival drop significantly. Therefore, there is an urgent need to find a drug therapy that not only affects the primary tumor, but perhaps more importantly, prevents the formation of metastases in the liver. Neuropilin 2 (Nrp2) is a transmembrane co‐receptor protein that binds a secreted protein called semaphorin 3F (S3F). Nrp2 is not expressed in the liver but is highly overexpressed in PDAC cells and PDAC‐associated endothelial cells. Based on these observations, we hypothesized that S3F expression in the liver could repel metastasizing PDAC cells. As part of a series of prevention trials, we performed an IV injection of S3F expressing adenovirus in mice and thereby induced the expression of the protein in the liver. Three days later we performed an orthotopic injection of syngeneic PDAC cells. For our intervention trials, we inverted the sequence of tumor and virus injections. Our results showed a significant reduction in tumor metastases in the experimental SEMA3F‐treated group compared to control virus treated mice. But the actions of Nrp2 are not limited to its ability to mediate repulsion. The tumor microvessel density and hence the overall PDAC tumor size was also reduced, thereby illustrating the systemic anti‐angiogenic effects of S3F‐mediated signaling via Nrp2. The action of Nrp2 on tumor growth was further elucidated by repeating the same experimental procedure in Nrp2‐deficient mice. Again, we saw a reduction in size and microvessel density within the primary tumor, which validates our initial hypothesis that Nrp2 is necessary for tumor growth and angiogenesis in PDAC. In summary the therapeutic effects of inhibiting Nrp2 are twofold: First and foremost, we have shown that S3F expression in the liver will repel Nrp2‐positive PDAC cells, as well as reduce tumor growth in the pancreas compared to the control. Finally, we have also demonstrated the anti‐angiogenic effects of S3F.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.