bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Role of Polyamines for Proliferation, Survival, and Infectivity of the Protozoan Parasite Leishmania donovani
Authors: Roberts Sigrid , Perdeh Jasmine , Abuan Irene , Le Linh , Love Quintin , LoGiudice Nicole , Turcu Kristen , Harrelson John 
Published: June 2021

A better understanding of parasite biology and host‐parasite interactions is critical for the development of much needed new therapeutic strategies against the neglected tropical disease leishmaniasis, which affects an estimated 12 million people annually. Polyamines are metabolites that play central roles in the biology of all eukaryotes, and recent studies have highlighted their critical nature in Leishmania parasites. In Leishmania, the polyamine biosynthetic pathway consists of four enzymes: arginase (ARG), ornithine decarboxylase (ODC), spermidine synthase (SPD), and S‐adenosylmethionine decarboxylase (ADOMETDC). These enzymes sequentially generate ornithine, putrescine and spermidine. We have generated a complete set of gene deletion mutants in Leishmania donovani (Δarg, Δodc, LdΔspd, and Δadometdc), in order to evaluate the polyamine biosynthetic enzymes as therapeutic targets and to study the functions of polyamines. In vivo studies revealed surprising differences in infectivity phenotypes, with the Δodc deletion exhibiting the most profound reduction in infectivity. In vitro studies showed that the gene deletion mutants depend on the provision of downstream metabolites to survive. An evaluation of cell growth between the different cell lines uncovered that the only vital role of ornithine is as precursor for polyamine synthesis. However, putrescine, which has previously been postulated to be merely a substrate for spermidine formation, has additional essential functions. Our studies revealed that putrescine is especially important for replication and proliferation. Both putrescine and spermidine were essential for cell survival, but the provision of either polyamine alone allowed parasites to enter a quiescent‐like state for several weeks. Future studies utilizing the gene deletion mutants will continue to elucidate the functions of polyamines for cellular proliferation, survival, and persistence.Support or Funding InformationNational Institute of Allergy and Infectious Disease Grant AI041622This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chai Tea Promotes Ampicillin Susceptibility in MRSA
Authors: Hinsdale Shannon , Pace Joel , Anderson Emeli , Favoreto Silvio 
Published: June 2021

Staphylococcus aureus (S. aureus) is a pathogenic organism that asymptomatically resides on the skin and in nasal mucosal membranes of 11–40% of the worldwide population. S. aureus has mutated in response to long‐term antibiotic exposure, generating strains known as methicillin‐resistant Staphylococcus aureus (MRSA). MRSA is involved in approximately 80,000 infections, coupled with 11,000 deaths per year in the United States alone; costing the U. S. healthcare system up to $9.7billion, or approximately $60,000 per patient, annually. Consequently, MRSA infections require stronger, more expensive drugs, accompanied by extended hospitalization. Treatment options are dwindling as costs are rising, both in mortality/morbidity rates and financially. Antimicrobial resistance is an ongoing process that will require alternative and creative interventions.Masala chai tea (chai tea) is a traditional tea beverage originating from India. This tea beverage is composed of a strong black tea base infused with various spices; typically cinnamon, cloves, cardamom, ginger, and black peppercorn. Each constituent listed above is composed of at least one bioactive plant compound with proven antimicrobial activity. We have taken these antimicrobial properties of each component into account, thus opting to use chai tea as a means to modify the resistance of MRSA to beta‐lactam antibiotics.In this study, we utilized seven MRSA strains. The microbes were isolated from carriers at the Cuesta College Allied Health program and were characterized by culture and PCR properties. Aqueous based chai infusion (CW) or ethanol‐based chai extract (CA) was incorporated into the media used for antibiogram diffusion assays, according to standard protocols. Compared to the control conditions, the aqueous‐based chai infusion and ethanol extract promoted a significant increase in MRSA susceptibility to Ampicillin, as demonstrated by the diameter of the inhibition halo (ANOVA, F=4.92, p<0.05).The results indicate that aqueous based chai infusion or ethanol‐based chai extract potentiated the bacteriocidal effect of Ampicillin against all seven isolates of MRSA in vitro. Interestingly, only Ampicillin, from all tested antibiotics, showed synergistic activity with chai. The data suggest the potential benefits from the association of established antimicrobial drugs with natural plant compounds as a means of modifying drug resistance in MRSA. The synergism shown between chai tea and ampicillin holds the possibility of assisting in deconstructing mechanisms of resistance, helping stage for future studies to progress towards reducing the MRSA global crisis.Support or Funding InformationWork supported by Cuesta College FoundationShannon Hill ‐ Advancement Foundation Executive Directorshannon_hill@cuesta.eduThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Benzimidazole Carboxamide Derivative, DDB‐506 Inhibits Bacterial Proliferation by Targeting FtsZ
Authors: TIWARI RISHU , DHAMELIYA TEJAS M. , CHAKRABORTY ASIT K. , PANDA DULAL 
Published: June 2021

FtsZ, a conserved cell division protein in prokaryotes has emerged as an attractive target for antibacterial drugs. FtsZ plays a key role in bacterial cell division; therefore targeting FtsZ assembly may be an efficient approach to inhibit bacterial cell division. In this work, we have checked the effect of several benzimidazole carboxamide derivatives on the bacterial cell proliferation. A benzimidazole carboxamide derivative, DDB‐506 was found to inhibit the proliferation of Bacillus subtilis (B. subtilis) with an MIC of 6 μM. DDB‐506 increased the length of B. subtilis cells by six times indicating that it inhibits bacterial cytokinesis. Light scattering, sedimentation and electron microscopic analysis of the effects of DDB‐506 on the assembly of Bacillus subtilis FtsZ (BsFtsZ) indicated that DDB‐506 inhibits the polymerization of BsFtsZ in vitro. DDB‐506 also inhibited the GTPase activity of BsFtsZ. Molecular docking analysis showed that DDB‐506 possibly binds to the nucleotide binding site of BsFtsZ. DDB‐506 also inhibited the binding of MANT‐GTP, a fluorescent analogue of GTP to FtsZ suggesting that the compound may bind to the GTP binding site of FtsZ.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Computational Studies of the NUDIX Hydrolase Superfamily
Authors: Richman Spencer Chase , O'Donovan Kevin , Craig Paul , Mills Jeffrey , O'Handley Suzanne 
Published: June 2021

The Protein Structure Initiative (PSI) was an effort by consortiums to solve as many unique protein structures as possible; the Protein Data Bank contains a number of enzymes whose structures have been solved, but for which no enzymatic activity has been determined. The Enzyme Function Initiative (EFI) is an effort to determine as many unique enzyme functions as possible. There are a number of putative NUDIX Hydrolase superfamily members for which structures have been solved, but for which no enzymatic activity has been determined. We have catalogued the structurally‐determined enzymes within the NUDIX Hydrolase superfamily using the PDB, BLAST, Dali, and SCOP. We are now in the process of characterizing these enzymes in the biochemistry teaching laboratory.Computational programs, such as ProMOL and UCSF Chimera, aid in annotating uncharacterized enzymes. ProMOL works by aligning the relative spatial arrangement of the catalytic residues of a reference enzyme with query searches. Chimera is a molecular visualization program that provides various tools for the structural analysis of proteins, including comparative modeling and conformational analysis. We are using these programs to design catalytic motifs for the NUDIX hydrolase superfamily and subfamilies within, which then can be used to uncover family members whose structures have been solved. Likewise, we have used the docking program PyRx, which utilizes AutoDock Vina, to predict substrate specificity, and have discovered that due to its “static” nature, it does not satisfactorily predict substrate specificity. Thus, next we will use a “dynamic” docking program to reevaluate substrate specificity prediction. Through this process, we have discovered the strengths and limitations of these programs.Bioinformatics tools, including databases and computational programs, are good starting points for determining protein function; however, the only way to definitively determine protein function is through experimentally determining activity. Our ultimate goal is to discover and characterize new enzymes from human pathogens as potential novel antibiotic targets.Support or Funding InformationNSF IUSE grant (PAC, JLM, and SOH). RIT College of Science FEAD grant (SOH and JLM).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery of Novel Small Molecule Inhibitors of Bacterial Pyruvate Carboxylase
Authors: Wyatt Brittney , Burkett Daniel , Mews Mallory , Donaldson William , Dockendorff Christopher , St. Maurice Martin 
Published: June 2021

Staphylococcus aureus and Listeria monocytogenes are pathogenic bacteria that can cause serious infections for immunocompromised individuals1, 2. Bacterial metabolism has been linked to pathogenicity and, in particular, the metabolic enzyme pyruvate carboxylase (PC) has been shown to be an important contributor to L. monocytogenes virulence1, 2. Small molecules that could inhibit bacterial PC activity would better reveal the role of this enzyme in a wide range of bacterial infections and could have the potential to be developed into antimicrobial agents. However, there are currently no known specific or potent small molecule inhibitors of PC activity3. We aim to find, develop, and characterize novel small molecule inhibitors of PC that will aid in understanding the structure, mechanism, and regulation of this enzyme. PC catalyzes the conversion of pyruvate to oxaloacetate (OAA) and a novel fixed‐time assay has been developed to detect OAA with a diazonium salt, Fast Violet B (FVB). The validated assay has been used to screen specific subsets of rationally designed inhibitors as well as compounds in an unbiased high‐throughput screen. Using both approaches, we have discovered novel small molecule inhibitors of bacterial PC that have the potential to be developed into valuable research tools and future antimicrobial agents.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 GENE EXPRESSION PROFILE AND TETRASPANIN PROTEIN CO029 ON THE HUMAN COLORRETAL CANCER LATERALITY
Authors: Braga Lucélia Antunes , Assis Jéssica Vieira , Moraes Vanessa Silva , Silva Istéfani Dayse , Grenfell Rafaella Fortini 
Published: June 2021

Colorectal cancer (CRC) is one of the most prevalent neoplasms and presents a multifactorial and complex etiopathogenesis, not yet fully elucidated. Studies have shown that tumors of the right colon differ significantly from neoplasms of the left colon, because they present greater risks of metastasis, are less responsive and exhibit different predictive and prognostic markers. Some of these markers have been studied, such as the tetraspanin CO029 protein, which favors cell motility, and epithelial‐mesenchymal (EMT) and neoplastic stem cell (CSC) markers, which are associated with loss of cell differentiation. According to studies, the three markers cited are closely associated with tumor progression. It has already been observed that in RCC tumor cells these markers present a high expression. Therefore, the analysis of the expression profile of tetraspanin CO029 and other markers in the RCC may become potential predictive tools for this neoplasia. Thus, the objective of the study is to evaluate the gene and protein expression of tetraspanin CO029 and correlate with laterality of RCC and with EMT and CSC markers. Therefore, it is proposed the use of human samples affected by the neoplasia proposed in different histopathological stages, for the evaluation of the immunohistochemical expression of tetraspanin CO029 protein. In addition, we will evaluate the gene expression of the three mentioned markers through the extraction of messenger RNA (mRNA) from these samples, for quantitative PCR (qRT‐PCR). The clinical data of the patients will be used for correlation with the expression of the proposed markers. Currently our efforts are concentrated in the evaluation by qRT‐PCR of the samples affected by CCR, where we observed that the samples located in the right colon presents a greater expression of tetraspanin CO029, corroborating with works that associate this protein with intensification of metastasis, due to the function of give motility to the cell. We are jointly conducting the production of anti‐tetraspanin CO029 monoclonal antibodies by in vitro methodology, which will be used for immunohistochemical labeling and in future projects, tested as monoclonal therapy. The production is in the phase of selection of positive hybridomas. Considering that the number of RCC cases is increasing in Brazil and in the world, it is important to carry out studies to elucidate this neoplasia. This project is relevant when we refer to possible predictive markers and future therapeutic targets, as it may make treatment personalized and possibly more efficient. Financial support: Fiocruz, CNPq, FIOCÂNCER IRR.Support or Funding InformationFinancial support: Fiocruz, CNPq, FIOCÂNCER IRR.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Understanding the Impact of IDH2 Mutations on the Redox Balance of Cancer Cells
Authors: Gelman Susan J. , McKenzie Lisa , Chheda Milan G. , Patti Gary J. 
Published: June 2021

Study ObjectiveWe seek to answer the following question: how does consumption of NADPH by mutant IDH2 for 2‐HG synthesis affect other NADPH‐dependent pathways in the mitochondria?MethodsHCT116 IDH2 mutants were incubated in the presence of H2O2 or irradiated with 3,6, or 9 Gy ionizing radiation (IR) and cell viability was assessed with a trypan blue exclusion assay. Stable isotope tracing was performed by incubating HCT116 cells (n=3) with the specified isotope for 12–24 hours, then harvested in ice‐cold methanol. After extracting metabolites from cells in a mixture of methanol, acetonitrile, and water, samples were run on a Thermo QExactive Plus using liquid chromatography/mass spectrometry (LC/MS) with a HILIC column in negative mode.Results and ConclusionIsocitrate dehydrogenase 2 (IDH2) mutants are found in cases of acute myeloid leukemia (AML), glioblastoma, chondrosarcoma, and lymphoma cancers. IDH2 is one of two nicotinamide adenine dinucleotide (NADPH)‐dependent isoforms of IDH. While IDH1 is found exclusively in the cytosol, IDH2 is localized to the mitochondria. Both enzymes are mutated in cancer to produce 2‐hydroxyglutarate (2‐HG) in millimolar amounts. The neomorphic reaction consumes the essential redox factor NADPH. Previous research indicates that IDH1 mutants are dependent upon upregulation of the pentose phosphate pathway (PPP) to supply more NADPH for 2‐HG synthesis. However, this pathway is localized to the cytosol and NADPH is not capable of crossing the mitochondrial membrane to supply the IDH2 reaction. To study the effects of NADPH consumption/production with respect to compartmentalization, we used stable isotope labeling and LC/MS to assess central carbon pathway flux in comparison to IDH1 and WT cells. Our preliminary data suggests that IDH2 mutants may be dependent on compensatory mechanisms to generate essential metabolic factors (such as ME1 and the folate cycle). We conclude that IDH2 mutations lead to metabolic rewiring of the TCA cycle to support the demand of 2‐HG synthesis in the mitochondria.Support or Funding InformationGJP received financial support for this work from the National Institutes of Health Grants R35ES028365 and R21CA191097 as well as the Alfred P. Sloan Foundation, the Pew Scholars Program in the Biomedical Sciences, and the Edward Mallinckrodt, Jr Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Profiling Bis(monoacylglycero)phosphate Lipids in Cancer Cell Lysosomes as Therapeutic Targets
Authors: Showalter Megan , Sa Michael , Tsugawa Hiroshi , Berg Anastasia , VanderVorst Kacey , Kind Tobias , Carraway Kermit L. , Fiehn Oliver 
Published: June 2021

BackgroundTumor resistance to apoptosis underlies the clinical failure of conventional chemotherapeutics and commonly employed molecularly targeted drugs. Cancer cells frequently activate survival pathways to evade destruction by apoptosis‐inducing therapies and persist following treatment to seed tumor recurrence and metastatic outgrowth. Lysosomal cell death is an alternative death pathway that triggers non‐apoptotic mechanisms. The lysosome, a major degradative component of eukaryotic cells, is altered by oncogenic transformation. Key to the function and stability of lysosome‐associated enzymes are bis(monoacylglycero)‐phosphate lipids (BMP). BMPs are negatively charged and highly enriched in the internal lysosome membrane. Untargeted analytical methods for the measurement of BMPs and in silico libraries to support the discovery of new BMP species are currently lacking.MethodWe present here an expanded BMP library to include over 1800 BMP species in both positive and negative ESI modes. We have chromatographic separation of BMP from isobaric species, and have used this method to profile cancerous and non‐cancerous immortalized cell lines. Lipidomic analysis was performed using Waters Acquity UPLC CSH C18 (100 mm length × 2.1 mm id; 1.7 μm) column coupled to a Thermo Scientific Q‐Exactive HF mass spectrometer. Data processing was done using MS‐DIAL software and in‐silico structure predictions were done using MS‐FINDER.ResultsIn breast cancer cells (Met‐1), we found that all BMP species measured were significantly less abundant relative to non‐transformed mammary epithelial cells (nMUMG), (n = 10). As BMPs are key to lysosome stability, we hypothesize the dramatic loss of BMPs observed in transformed cells renders cancer‐associated lysosomes unstable. Therefore, targeting BMPs might represent an effective therapeutic strategy for selective cancer cell destruction.Support or Funding InformationThis work was funded by NIH U24 DK097154.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Herbal Mixture Extracts on Degenerative Arthritis In Vivo Models
Authors: Moon Yun Jeong , Dasom Lee , Dakyung Kim , Minhee Lee , Sujeung Park , Yongwook Lee , Junkee Hong , Jeongmin Lee 
Published: June 2021

This study was to investigate the effects of herbal mixture extract complex (S1, S2) on the monosodium iodoacetate (MIA)‐induced osteoarthritis in rats. Arthritis was induced by injection of MIA (6 mg/m L) into knee joints of rats. Arthritic rats were divided into control and treated (S1L, S1H, S2L and S2H) group. Control group was taken AIN93G diet and treated group was taken samples (S1L, S1H, S2L and S2H) by orally for 21 days. Body weights were measured at 0, 7, 14, 21 days after MIA injection. At the end of experiment (21day after MIA injection), gross and histopathological examination on the articular structures of knee joints were performed. Prostaglandin E2 (PGE2) contents in serum were measured by ELISA method. TNF‐α, IL‐1β, COX2 and, NF‐kB contents in chondrocyte were measured by Real‐time PCR method. And also, proteoglycan aggregate (Aggrecan), collagen type 1,2,10, TIMP‐1, MMP‐3 and MMP‐7 content in chondrocyte were measured by Real‐time PCR method.PGE2 contents in serum level of the treated group were decreased compared with control group. TNF‐α, COX2 and IL‐1β contents in chondrocyte of the treated group were significantly decreased compared with control group. NF‐kB contents in chondrocyte of the S2H treated group were significantly decreased compared with control group. Collagen type 1 and 2 contents in chondrocyte of the treated group were significantly increased compared with control group. MMP‐3 and MMP‐7 contents in chondrocyte of the S2H treated group were significantly decreased compared with control group. The results of the histopathology showed that the severity of the arthritis was slightly reduced compared to the control group.On the basis of these results, we concluded that S1 and S2 treatment had anti‐arthritic effects on the monosodium iodoacetate‐induced osteoarthritis in rats.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Improvement of joint health by anti‐inflammatory activity of natural substances in chondrocyte of osteoarthritis
Authors: Kim Dakyung , Lee Minhee , Park Soo‐Jeung , Yun Jeong‐Moon , Lee Dasom , Sohn Johann , Yun Min‐Kyu , Lee Jeongmin 
Published: June 2021

Osteoarthritis (OA) is a chronic degenerative disease that involves inflammation and pain due to damage or degenerative changes in the cartilage that protects the joints. OA has a high incidence in the elderly. Treatment methods include relieving symptoms and preventing cartilage damage. Drugs used to treat osteoarthritis have side effects and it is important to reduce them. Therefore, it is necessary to develop a therapeutic agent for osteoarthritis using natural substances. The purpose of this experiment was the anti‐inflammation effect of natural substances on osteoarthritis in primary cultured rat cartilage chondrocytes. Aggrecan, collagen type I, □, □, MMP‐3, 7, NK‐kB and COX‐2 expression were measured by real‐time PCR. TNF‐α and IL‐1β secretion was measured by ELISA kit. H2O2 and LPS‐induced chondrocytes were used to measure concentrations of 100, 200, and 400 (μg/ml). As a result, S400 significantly increased the gene expression level of aggrecan, collagen type I, II, III and decreased gene expression of pro‐inflammatory factors NF‐kB, COX‐2 and MMPs (3 and 7). The cytokine productions of TNF‐α and IL‐1β also decreased in S400. Therefore, S400 had an anti‐inflammatory effect and alleviated the inflammation occurs in OA.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.