bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison study of exosomes molecules driven from (NCI1975) NSCLC cell culture supernatant isolation and characterization techniques
Authors: Mahgoub Elham Omer , Haik Yosef , Qadri Shahnaz 
Published: June 2021

BackgroundLung cancer consider as one of leading cause of death around the world, normally the disease detection is difficult in early stages, Anew methods for diagnostic using non‐invasive way are developed using Nano‐molecules called exosomes. Exosomes are cell derived vesicles carrying different kind of molecules (proteins, DNA, RNA, microRNA) and also, displaying many proteins on their membrane surfaces. Those exosomes molecules are identical of parental cancer cells that induce them.MethodsIn this study, exosomes were isolated and characterized using multiple centrifugations, ultrafiltration, and chemical precipitation techniques. The resulted products were captured by two electro‐Microscopes (EM) methods for characterizing the morphology of exosomes as to differentiate exosomes from other extracellular vesicles Besides, the resulted products also further characterized by blotting the proteins extracted from exosomes against different monoclonal antibodies (CD81, CD63, CD9) that confirmed the existence of ligands attached to exosomes surfaces in western blot test.Results and Discussionthe exosomes isolated products were screened under Transmission Electron Microscopy (TEM). The size distributions of extracellular vesicles were significantly different from exosomes 160 nm P < 0.001). The size distributions of apoptotic vesicles (APV) 450nm and necrotic bodies, (NCB) 280 nm were significantly different from exosomes (P < 0.001). The same size band was observed in PVDF membranes of western blot test in all three methods using each monoclonal antibody. However, the filtration devices give higher protein yield and greater purity.In ConclusionsThe current results confirmed the accessible use of exosomes as biomarker to detect lung cancer cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antiproliferative Properties of Lichens Parmelia vagans and Parmelia sulcata
Authors: Bondarenko Vladimir , Raymond Ky 
Published: June 2021

The purpose of this project was to analyze antiproliferative properties of the secondary metabolites extracted from lichens Parmelia vagans and Parmelia sulcata collected from the semi‐deserts of Russia. An isolation and characterization of novel natural compounds that can selectively slow down or stop proliferation of cancer cells can enhance development of anticancer therapeutics.Various lichen extracts were prepared using either water or organic solvents. We assessed the antiproliferative activity against several human cancer cell lines; human hepatocellular carcinoma (Hep G2), human non‐small cell lung cancer (NCI‐H1299), and human mammary gland ductal carcinoma (T‐47D). The same number of cells of different cell lines were plated into a 96‐well plate and allowed to adhere overnight. Next day, different dilutions of the lichen extracts were added to the adhered cells and the plate was placed in CO2 incubator for 6–72 hours. By the end of the incubation time a picture of each well was taken to assess cell confluency and the cells were subjected to Resazurin assay to analyze the effect of the lichen extracts on cell growth and proliferation. After Resazurin assay, the cells in each well were treated with trypsin, suspended in growth medium and counted using Bio‐Rad cell counter to evaluate a percent of live cells.Our experimental data from cell viability assay and live cells count indicate that bottled water and ethyl acetate extracts of P. vagans and P. sulcata significantly suppress (30%–60%) the growth of human hepatocellular carcinoma and human non‐small cell lung carcinoma. We also found that cancer cells treated by the lichen extracts results in their morphological changes that resemble an activation of apoptosis.Our findings indicate that these lichen species produce secondary metabolites that slow cancer cells growth and proliferation. Future experiments will be focused on isolation and characterization of the compounds from P. vagans and P. sulcata crude extracts that are responsible for the inhibition of cancer cells growth. Another important direction of the future research is to identify the cellular target(s) of the antiproliferative compound(s) and its molecular mechanism of action. I summary, the lichen's secondary metabolites may hold a vast medicinal potential and could be a viable source of a novel of anticancer drugs.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
UPLC‐MS Analysis of Characterization of Bioactive Compounds Produced by Endophytic Bacillus tequilensis ALR‐2 from Aloe vera Plant
Authors: Akinsanya Mushafau Adewale , Goh Joo Kheng , Ting Adeline Su Yien 
Published: June 2021

Endophytic Bacillus tequilensis ALR‐2 isolated from Aloe vera plant, was found to have antimicrobial activities attributed to a series of lipopeptide antibiotics and biosurfactants produced. Partial‐purification of its ethylacetate extract by column chromatography gave SF5 (5mg mL−1) which effectively inhibited Bacillus cereus and Staphylococcus aureus (28.6±0.5 and 30.3±0.5 mm zone of inhibition, respectively), and significantly more effective than the standard antibiotics Ciprofloxacin (5μg) (23.6 ±0.5 and 18.6 ±0.5 mm, respectively) and Kanamycin (30μg) (20.3 ±0.5 and 13.6 ±0.5 mm, respectively) against the two pathogens. UPLC‐MS analysis of SF5 showed three series of ion peaks. The quadrupole‐orthogonal ionization time‐of‐flight mass spectrometer revealed [M+H]+ of 678.4601; 679.4690 and 680.5076 were shown to correspond to the oligopeptide antibiotics, monamycin (mol. wt. 677.84) and [M+H]+ of 1119.8870; 1120.8992 and 1121.9119 correspond to antibiotic complex 61–26 (mol. wt. 1120.348) a lipopeptide. However, [M+H]+ of 1356.0151 and 1358.0664 could not be classified to what is known in the literatures.Support or Funding InformationThis work was supported by Higher Degree for Research (HDR) School of Science, Monash University Malaysia and Sponsorship from Tertiary Education Trust Fund of Federal Republic of Nigeria. We wish to acknowledge the assistance provided by Ms Amreeta Sarjit of Microbiology Teaching Laboratory of Monash University Malaysia for the provision of bacteria pathogens used for this project.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An Investigation of Milly, a K2 Mycobacteriophage
Authors: Conte Amy , Ganey John , Cornely Kathleen 
Published: June 2021

Mycobacteriophages are viruses that infect bacteria of the genus Mycobacterium. These phages are capable of undergoing two life cycles: the lytic cycle and the lysogenic cycle. In the lytic cycle, the phage inserts its DNA into a bacterial host, and in response, the host produces all of the proteins and other materials necessary to create more phage. The phage then multiply exponentially within the bacterial host until the bacterium lyses, releasing thousands of new phage to infect more bacteria. On the other hand, during lysogeny, the bacterium incorporates the phage DNA into its own genome, allowing it to replicate without producing new phage or lysing. Due to the increasing prevalence of antibiotic resistance, we are interested in taking advantage of the lytic cycle in order to treat bacterial infections, such as tuberculosis, with mycobacteriophage therapies. Using the Bacteriophage Recombineering of Electroporated DNA (BRED) method, we hypothesized that we could delete the gene coding for the repressor protein on Milly's genome, which would prevent Milly from undergoing lysogeny and make it lytic‐only. Here we report a characterization and manipulation of the K2 cluster mycobacteriophage Milly, isolated at Georgia College and State University, which we have determined to be capable of both the lytic and lysogenic phage life cycles. We are in the process of demonstrating the deletion of Milly's repressor gene so that it loses its ability to undergo lysogeny. This way, we can utilize Milly's lytic cycle as a way to infect and kill disease‐causing bacteria.Support or Funding InformationThis research is funded by the Providence College Undergraduate Research Committee. We would also like to thank the Providence College Department of Chemistry and Biochemistry for their support.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Step Past Phage Hunters, Modification of the Mycobacteriophage ZoeJ Genome for Phage Therapies
Authors: Scano Colin J , Jancevski Alicia , Scola Shelby , Ferrante Aaron 
Published: June 2021

Mycobacteriophages are viruses that infect host mycobacterial cells like the mycobacterium tuberculosis. When infected, the host bacterial cell will either enter a lytic or lysogenic phase. In the lytic cycle, the host cell is transformed into a phage DNA producing factory. This host cell then lyses, releasing numerous new phages that infect other bacterial cells, carrying on the cycle. During lysogeny, however, the phage genome is integrated into the bacterial cell genome and the host cell does not lyse, remaining dormant for periods of time. The use of phages for the treatment of bacterial infections known as phage therapy is an alternative to the conventional use of antibiotics and could potentially replace it entirely. Since only the lytic cycle offers antimicrobial treatment, problems arise in phage therapy due to the inability to control the lifecycle of phages. A solution to this problem involves the creation of a lytic‐only phage through alterations of the phage genome. The phage genes responsible for the lysogenic cycle and genome integrations are the repressor and integrase genes. The goal of this project is to alter the phage genome in order to construct lytic‐only phage that lacks the repressor or integrase genes. This project focuses on the K2 cluster phage ZoeJ that was discovered in the SEA‐PHAGES/Phage Hunters program at Providence College. We have previously demonstrated that the repressor gene can be knocked out in the ZoeJ phage, and now aim to knock out the integrase gene in the wild‐type ZoeJ phage, as well as in the Δgp46 ZoeJ phage. Using the BRED method developed by the Hatfull laboratory, we aim to take advantage of the homologous recombination of bacteria DNA with phage DNA to delete the integrase gene from the ZoeJ phage genome.Support or Funding InformationFunding for the project and trip has been supplied by the Providence College Biochemistry department through the PC small grants for undergraduate research.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Demonstration of Photodynamic Molecules as Larvicides towards the Yellow Fever Mosquito Aedes aegypti
Authors: Meier Cole , Rouhier Matthew 
Published: June 2021

Each year, mosquitoes serve as a vector to transmit diseases that kill upwards of 750,000 people. Although the need for vector control of these organisms is clear, the misuse of modern pesticides has caused significant ecological damage as well as the development of pesticide‐resistant mosquitoes. Photosensitive insecticide technology has shown potential to minimize ecological damage by targeting classically resistant mosquitoes in new ways. These photoactive molecules, specifically safranin‐O (Saf‐O) and methylene blue (MB), transfer their energy to molecular oxygen to form reactive oxygen species (ROS). These species damage biological systems in the mosquito that differ from the systems excessively targeted by modern pesticides. By targeting new and less resistant biological systems, photosensitive insecticides present a novel strategy for vector control. Currently the literature lacks the in vivo pathway describing ROS generation by photodynamic insecticides. Here we show the generation of ROS in vivo using Saf‐O and MB along with a model of the photodynamic interaction with the Aedes aegypti larva.Support or Funding InformationThe authors would like to thank the Kenyon College Summer Scholars Program for the funding received to pursue this project.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluating the Chemistry and Biological Action of Midwestern Medicinal Plants
Authors: Nguyen Nguyen‐Anh Thi , Shima Rafaella Tami , Abelkis Alexandra , Carter Jasmine , Cornwell Claire , Scerbak Courtney , Deibel Michael 
Published: June 2021

Literature has demonstrated Caenorhabditis elegans possess similar cellular mechanisms when responding to toxic stress and increasing lifespan. In addition, a plant‐based diet with antioxidant activity and high phenolic content are associated with increased health. We tested the hypothesis that medicinal plant extracts would improve C. elegans survival rates when exposed to toxic (hot and cold temperature) stresses. The chemical content and biological activity of mulberry, goldenrod root, goldenrod leaf, blue violet, black raspberry, and red root plant extracts were measured and their compound class fractions (anthocyanin, flavonol, tannin, and phenolic acid). We found that with increased phenolic content, extracts had higher antioxidant activity; this correlation is largely attributed to anthocyanin content. Additionally, phytochemicals not isolated in our fractions and/or synergistic interactions contributed to the overall antioxidant capacity of extracts. We did not find a correlation between antioxidant capacities and C. elegans survival rate under toxic stresses. However, mulberry (50 μg/mL) significantly increased the survival rate in response to cold stress and blue violet ( 400 μg/mL) extract treatment trended towards increased survival rate in heat stress. This research is a preliminary step for further study and discussion to understand the comparison between medicinal plant activity and C. elegans survival and longevity.Support or Funding InformationThis work was funded by the Earlham College Center for Global Health and a Senior Research Grant from the Indiana Academy of Sciences (to Courtney Scerbak).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of oxidized mitochondrial mRNA in human neurons under oxidative stress
Authors: Rana Joram , Kharel Prakash , Basu Soumitra 
Published: June 2021

Mitochondrial dysfunction and the overproduction of reactive oxygen species (ROS) are key features of many neurological disorders. Electrons leaking from the electron transport chain (ETC) in the mitochondria reduces molecular oxygen to superoxide, which through enzymatic and non‐enzymatic reactions generates ROS. Under normal physiological conditions ROS are used in cellular homeostasis and cell signaling, but during stress conditions, they are often overproduced. ROS are known to damage biomolecules DNA, RNA, proteins, and lipids. RNA damage has received attention in recent years. Studies in our lab demonstrated that the selective oxidative damage to RNA could lead to altered gene expression and differential production of protein, contributing to neurodegeneration in multiple sclerosis (MS). The mitochondria contain its own DNA, which transcribes into RNA and subsequently gets translated into mitochondrial proteins using its unique machinery. Selective mitochondrial RNA oxidation could disrupt this process, damaging the mitochondrial respiratory chain proteins, and eventually causing more mitochondrial dysfunction, contributing towards the progression of neurodegeneration in MS and other neurological disorders. The SH‐SY5Y human neuronal cells were treated with sodium nitroprusside (SNP, NO·/·OH) to induce stress and generate ROS to mimic the diseased condition. By quantitating cellular RNA, we have displayed the differential mRNA expression of a number of genes during oxidative stress, for example, high expression of superoxide dismutase 1 and carbonyl reductase. In this study, we focused on mRNA transcribed specifically inside the mitochondria to examine which sequences are most susceptible to oxidation by ROS. We found that 9 of the 13 mRNAs transcribed by mitochondria are highly oxidized. The oxidation of these mitochondrial mRNAs might lead to complication in protein synthesis and might play a role in mitochondrial dysfunction.Support or Funding InformationDivision of Research & Sponsored Programs (SURE program), Kent State University Graduate Student Senate, Kent State University to PK.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synergistic Chemopreventive Effect of Medicinal Plants against Azoxymethane‐induced Oxidative Stress and Colon Carcinogenesis
Authors: Waly Mostafa I 
Published: June 2021

BackgroundOxidative stress plays an essential role in the development of different types of cancer malignancy, including colon cancer. The protective effect of different medicinal plants against oxidative stress‐mediated colon carcinogenesis has been reported, yet their synergistic effect was not elucidated.ObjectiveThis study investigated the antioxidant potential properties of a mixture of five medicinal plants (Rumex vesicarius, Oxalis corniculate, Pteropyrum scoparium, Moringa peregrine, and Caralluma arabica) in a rat model of colon cancer.MethodsAzoxymethane (AOM), a colon selective carcinogenic agent, was used to induce colon cancer in Sprague Dawley rats which were divided into 4 groups, n=10, (Control, AOM‐injected group and the other two groups were fed ad‐libitum for 12 weeks a diet supplemented with a mixture of the five different medicinal plants in the presence or absence of AOM injection). By the end of the experiment, all rats were sacrificed; colon tissues were excised and homogenized for biochemical measurements of: oxidative DNA damage, reduced glutathione, and total antioxidant capacity. Histopathological examination for colon tissues was also examined for aberrant crypt foci (ACF) development.ResultsAOM induced oxidative stress in the colonic milieu as evidenced by high level of oxidizing DNA, glutathione depletion, and impairment of cellular total antioxidant capacity. In addition, AOM increased the formation of ACF. Meanwhile rats fed with the five medicinal plants mixture showed a protective effect against the AOM‐induced oxidative insult and attenuated the colonic ACF formation.ConclusionThis study revealed that the examined medicinal plants provided a colon specific protection and might be used as a functional dietary bioactive agent in the context of primary prevention of colon cancer.Support or Funding InformationSultan Qaboos university Funding, Internal grant (IG/AGR/FOOD/01/17)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exposure to Electronic Cigarette Vapor Causes Systemic Accumulation of Biomolecular Free Radicals
Authors: Clemons Garrett , Aboaziza Eiman , Burrage Emily , O'Reilly Juliana , Chantler Paul , Kelley Eric , Olfert Mark 
Published: June 2021

Little is known regarding the impact of electronic cigarette (e‐cig) usage in the human body. However, emerging literature suggests that vaping results in endothelial cell mediated vascular impairments, altered thrombosis, and depressed immune function (similar to that observed with smoking). Since e‐cigs are marketed as a ‘safer’ alternative to traditional cigarettes, it is important to characterize mechanisms through which e‐cigs may induce cellular insult. It is known that traditional cigarette use results in systemic accumulation of biomolecular free radicals (BFR), thus we sought to determine whether exposure to e‐cig vapor will result in significant systemic increases in BFR. We hypothesize that vaping will increase the relative amount of BFR accumulation in vital tissues in a time‐dependent manner.To identify BFR in‐vivo, C57BL/7 mice were given IP 1.5 g/kg DMPO (5,5‐dimethyl‐l‐pyrroline‐l‐oxide) spin‐trap in 2 doses over 1 hour, and acutely exposed to french vanilla‐flavored e‐cig vapor (18 mg nicotine) for either 1 hour (n=3) or 4 hours (n=3). Sham mice were exposed to ambient air (n=3). Immediately following exposure, the mice were anesthetized, perfused with PBS via intra‐ventricular catheterization, organs/tissues were harvested and fixed in paraformaldehyde/sucrose, flash‐frozen in liquid nitrogen and then fixed in TFM. Imaging from fluorescent immunohistochemical labeling revealed significant increase BFR accumulation in the liver (223%, p<0.05) and to lesser extent in the heart (133%, p<0.05) in 1‐hour exposed animals. Similar trends for increased BFR (albeit not statistically significant) were found in the lung (236%) and brain (124%) in 4‐hour exposed group compared to air exposed controls.In summary, we found significant increase in the presence of BFR in the liver and heart of mice acutely exposed to 1‐hour of e‐cig vapor compared to controls, with similar trends in other tissue beds (lung, brain) after 4‐hours vape exposure. This is a pilot study which needs additional samples for further evaluation, but suggests that vaping induces BFR accumulation in multiple organs which could lead to cellular damage and tissue dysfunction.Support or Funding InformationSupport: WVU Cancer Institute Philip R Dino Innovative Research Grant (IMO); Transition Grant Support; Office of Research and Graduate Education (IMO), WVU Health Sciences Center APS STRIDE Fellowship (JO); NIHGMS 5U54GM104942‐03 (PDC)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Transcriptomics and Functional Genetics Approach to Identifying Components of the Drosophila Polyamine Transport System
Authors: Saldanha Joanne , Haney Michael , Landry Brianne , Figueroa Natalia , von Kalm Laurence 
Published: June 2021

Polyamines are essential growth factors for normal and malignant cells. Cells obtain polyamines via biosynthesis or transport from the extracellular environment. Polyamine depletion therapy is an attractive option to inhibit the growth of cancer cells. An FDA approved drug inhibits polyamine biosynthesis, however malignant cells typically respond by upregulating the polyamine transport system. Therefore, a dual therapy simultaneously targeting the biosynthetic and transport pathways is desirable. However, this approach is currently hampered by a poor understanding of the transport system in multicellular eukaryotes. To address this problem, we used RNA‐Seq to identify genes that are up‐ or down‐regulated under conditions where Drosophila larvae must transport polyamines to survive. From the screen 36 candidate genes for roles in polyamine transport were identified. We have also developed a functional genetic assay to test individual candidate genes. Our results suggest the existence of tissue‐specific polyamine transport systems, and post‐transcriptional regulation of polyamine transport.Support or Funding InformationUniversity of Central Florida, College of Sciences Seed AwardThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.