bluuebunny/crossref_metadata_2025_split huggingface dataset:
RNA helicase homolog and its association with the JAMM2 metalloprotease required for targeted protein turnover in Archaea
Authors: De Jesus Ramirez Cuper , Hauserman Matthew , Cao Shiyun , Maupin‐Furlow Julie A 
Published: June 2021

A functional analogue of the eukaryotic ubiquitin‐proteasome system (UPS) is found in archaea. This archaeal system, termed the SAMP‐proteasome system (SPS), operates in targeted protein turnover; however, many of its protein components [E1‐like UbaA, Cdc48/p97‐/Rpt‐ type AAA ATPases, and JAB1/MPN/MOV34 metalloenzyme (JAMM/MPN+) homologs] remain to be fully characterized. To dissect the role of the JAMM metalloprotease homologs within the SPS, we previously expressed and purified the two Haloferax volcanii JAMM/MPN+ homologs (HvJAMM1/2) from recombinant Escherichia coli. Only HvJAMM1 was demonstrated to be active as an Ubl isopeptidase. By contrast, HvJAMM2 was found to be required in vivo for protein turnover by the SPS. Here we focused on identifying and characterizing the protein partners and/or biochemical conditions which may be required in vitro for HvJAMM2 activity. For this objective, His6‐HvJAMM2 was expressed in H. volcanii and purified by Ni2+‐ chromatography to overcome factors that may be absent from recombinant E. coli. Co‐purifying protein partners were identified by LC‐MS/MS analysis, including HVO_2955 which is predicted to be a structural homolog of the JAMM/MPN− interacting RNA helicase Brr2 of the eukaryal spliceosome. While hvo_2955 was not essential for growth, we found an Δhvo_2955 mutant to have decreased growth under standard conditions, with the genetic defect trans complemented by expression of His6‐HVO_2955. Current work is focused on determining the role of HVO_2955 during stress as well as its potential binding to JAMM‐domain proteins in archaea. Assessing the role of this Brr2 structural homolog in RNA processing in conjunction with SPS components such as HvJAMM2 is anticipated to show a clearer picture of archaeal physiology.Support or Funding InformationFunds for this project were awarded to JM‐F through the Bilateral NSF/BIO‐BBSRC program (NSF 1642283), the U.S. Department of Energy, Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences and Biosciences, Physical Biosciences Program (DOE DEFG02‐05ER15650) and the National Institutes of Health (NIH R01 GM57498). Additional support is provided by the University of Florida's Preeminence Fellowship awarded to CR.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Conditional Protein Splicing of Inteins from Extremophiles
Authors: Mills Kenneth V 
Published: June 2021

Protein splicing is a post‐translational process facilitated by an intein, by which the intein catalyzes its own excision from flanking polypeptides (the exteins), concomitant with ligation of the exteins. Many inteins are themselves interrupted by homing endonuclease domains that can facilitate homing into intein‐less alleles. We have discovered inteins from extremophiles that can be expressed in E. coli, purified as unspliced precursors, and induced to splice by cues such as incubation under reducing conditions, at high temperature, or at high ionic strength. We have shown that the activity and folding of inteins from the extreme halophiles Halobacterium salinarum and Haloquadratum walsbyi demonstrate variable dependence on salt concentration, and we have designed transgenic organisms lacking inteins to test the physiological role of splicing. We have solved the structure of an intein from the extreme thermophile Pyrococcus abyssi, and studied the influence of structural elements on its stability at high temperature and pressure. We also have discovered that the splicing and nuclease activity of inteins from extreme thermophiles can be influenced by pressures consistent with those found at the seafloor.Support or Funding InformationThis work was supported by grants from the National Science Foundation (grants MCB1244089 and MCB1517138) and the Camille and Henry Dreyfus Foundation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using fluazifop‐p‐butyl for low cost increases in lipid accumulation for the generation of algal biofuels from Chlorella vulgaris
Authors: Pauley Annabella Grace , Smythers Amanda , Perry Nicole , Kolling Derrick 
Published: June 2021

The demand for sustainable, low‐emission fuels is increasing with the global expansion of industrialization. Biofuels have the potential to meet this need without the problematic addition of carbon into the earth's atmosphere. Algae, a potential biofuels feedstock, have the capability to quickly accumulate biomass under various conditions, including low‐nutritional growth medium and a reduced need for arable land. Current research is seeking to increase organismal efficiency in order to increase economic feasibility. Previous work has shown that exposing algae to environmental stressors or limitations, such as nitrogen deprivation, can increase lipid production in the cultures, though this is typically accompanied by a decrease in overall biomass. Nitrogen deprivation is also expensive to facilitate, so finding more economical methods is paramount to increasing algal biofuel feasibility. In this study, Chlorella vulgaris, a unicellular green alga, was exposed to a formulation of an inexpensive and readily available mass‐produced herbicide that targets acetyl Co‐A carboxylase with its active ingredient, fluazifop‐p‐butyl. The compositional changes of the algae cultures were monitored as well as the photosynthetic efficiency. Preliminary results have shown an increase in percent lipids in cultures exposed to the herbicide formulation, though the overall biomass decreased. Current results in this study will be presented.Support or Funding InformationThis research was made possible by NASA West Virginia Space Grant Consortium, Training Grant # NNX15AI01HThis material is based upon work supported by the National Science Foundation under Cooperative Agreement No. OIA‐1458952This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cytotoxic Interactions between Pseudomonas aeruginosa Virulence Factors and Metal‐Based Antimicrobials In Vitro
Authors: Mawhinney Thomas , Mossine Valeri V. , Chance Deborah L. , Waters James K. 
Published: June 2021

Emergence of multidrug‐resistant (MDR) bacterial pathogens has triggered a search for alternatives to current antibiotic‐based treatments. Toxic metal salts are widely used as anticeptics, but gallium(III) may act systemically, by disrupting iron metabolism in MDR pathogens like Pseudomonas aeruginosa. As P. aeruginosa infections release a range of toxins on their own, we asked whether bacterial toxins may interact with metal‐based drugs against the host.Using stress‐inducible transcription factor (Nrf2, HIF‐1α, ATF‐6, and MTF‐1) reporter constructs, we have developed a cell‐based protocol for simultaneous evaluation of the cellular metabolic activity, transcriptional/translational activity, and activation of intracellular stress responses in NIH‐3T3 fibroblasts and BEAS‐2B bronchial epithelial cells. The cytotoxicities of four P. aeruginosa cytotoxic virulence factors phenazine‐1‐carboxylate, phenazine‐1‐carboxamide, pyocyanin (Pyo) and its main metabolite, 1‐hydroxyphenazine (1HP), alone and in combinations with five clinically approved antimicrobial metal salts (Ag, Zn, Ce, Bi, and Ga) were evaluated in vitro.Bacterial phenazines Pyo and 1HP displayed cytotoxicities (IC50 30–90 μM) at concentrations comparable with those detected in sites of P. aeruginosa infections. Silver nitrate was the most cytotoxic metal salt (IC50 < 10 μM), while citrates of cerium(III) and bismuth(III) were not affecting cell proliferation even at 1 mM. In combinations, Bi and Ce inhibited cytotoxicity of Pyo and 1HP, increasing the IC50 2‐fold. In contrast, zinc(II) and gallium(III) strongly synergized with the phenazines, with a decrease in the IC50 reaching over 100‐fold in some experiments. Both Pyo and 1HP strongly (200–500‐fold) activated Nrf2 in the reporter cells, suggesting oxidative stress is a significant contributor to their cytotoxicity mechanism. Both Zn and Ga induced activity of MTF‐1, at subtoxic concentrations, thus implying that interaction of these metals with cytotoxic phenazines occurs along pathways maintaining intracellular redox homeostasis.This is the first report documenting cytotoxic interactions between clinically approved antimicrobial metal salts and bacterial phenazines at physiologically relevant concentrations. It provides a rationale for further mechanistic and in vivo studies of the phenomenon.Support or Funding InformationThis research was supported by the University of Missouri Agricultural Station Chemical Laboratories, the Cystic Fibrosis Association of Missouri (CFAM), the Leda J Sears Trust Fund for Child Health Research; and National Institute of Food and Agriculture (NIFA) grant No. MO‐HABC0002.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Macrophage migration inhibitory factor (MIF):A novel therapeutic target against aggressive breast cancer
Authors: Das Subhadip , Chatterjee Nabanita , Ahirwar Dinesh K , Mishra Sanjay , Varikuti Sanjay , Kaul Kirti , Satoskar Abhay R , Ganju Ramesh K 
Published: June 2021

Triple negative breast cancer (TNBC), defined as loss of estrogen, progesterone and Her2 receptors, is a subtype of breast cancer with worse prognosis and poor survival rate. Increased inflammation further reduces the TNBC patient survival. Macrophage migration inhibitory factor (MIF) is a pro‐inflammatory molecule known to enhance breast cancer progression and metastasis. However, not much is known about the role of MIF in TNBC progression and metastasis. MIF interacts with various oncogenic molecules and signaling pathways. Therefore, targeting MIF might be a promising strategy to inhibit TNBC progression and metastasis. Here we report that CPSI‐1306, a small molecule inhibitor specific to MIF, possesses the ability to target TNBC cell in vitro by inducing apoptosis. The mechanistic studies revealed that CPSI‐1306 induces apoptosis by enhancing reactive oxygen species (ROS), mitochondria membrane potential alteration, cytochrome C release and activation of Caspases. In addition, CPSI‐1306 also inhibits cell survival and apoptosis signaling molecules, including AKT, PDK and RAF expression. Next, we analyzed the clinical utility of CPSI‐1306 using mouse MVT‐1 cells derived orthotopic and human MDA‐MB‐231 cells derived xenograft TNBC mouse models. The oral treatment of tumor bearing mice with CPSI‐1306 significantly inhibited TNBC growth and metastasis to the lungs in a dose dependent manner. The histological analysis of tumors showed higher number of apoptotic cells in CPSI‐1306 treated tumors compared to vehicle controls. We also observed an association of higher MIF mRNA expression with poor patient survival, including TNBC, by using publically available datasets. These results suggest that CPSI‐1306 induces cell death in TNBC cells via activating ROS and mitochondria‐mediated apoptosis and significantly inhibits TNBC growth and metastasis in pre‐clinical TNBC mouse models.Support or Funding InformationDOD, Breakthrough AwardThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interrelationship between sex, age, blood volume and VO2max
Authors: Convertino Victor , Koons Natalie , Schlotman Taylor , Suresh Mithun 
Published: June 2021

BackgroundCirculating blood volume (BV) and maximal oxygen uptake (VO2max) are physiological characteristics important for optimal human performance in aerospace and military operational environments. Addressing the interrelationships between BV, VO2max, sex, and age together is relevant to identifying those individuals with greatest risk of poor outcomes from hemorrhage, as well as assessing the levels of performance competence of individuals deployed in operational environments. In the present investigation, we report and compare for the first time BV and VO2max in a cohort of males and females together to test the hypothesis that BV and VO2max is lower in older people independent of sex.MethodsTo accomplish this, a ‘data mining’ effort of an historic database generated from NASA and U.S. Air Force experiments was conducted. BV, red cell volume, plasma volume, hematocrit, and VO2max were measured in 94 healthy individuals (28 females, 66 males) across an age range of 23 to 65 years to assess the interrelationship between sex, age, BV, and VO2max. Subjects were classified in age groups by < 40 years and ≥ 40 years; these groups identified females as pre‐ vs. post‐menopausal.ResultsConsistent with our hypothesis, comparisons revealed that males had higher (p < 0.01) BV, red cell volume, hematocrit, and VO2max than females when standardized for body size. Against expectations, BV was not different in older compared with younger males and females. VO2max was not different in older compared with younger females, while VO2max was lower in older males (p < 0.01).ConclusionWe conclude that physiological mechanisms other than BV associated with aging appear to be responsible for a decline in VO2max of our older male subjects. Furthermore, factors other than menopause may also influence the control of BV in the female population. Our results provide evidence that aging may not compromise males or females in scenarios where BV can affect performance in aerospace and military environments.Support or Funding InformationThis research was supported in part by appointment of Ms. Koons to the Postgraduate Research Fellowship Program at the US Army Institute of Surgical Research administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the US Department of Energy and USAMRMC. Study funding was provided by a grant from the US Army Combat Casualty Care Research Program (D‐009‐2014‐USAISR).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Distribution and Expression of the FMRFamide‐gated Sodium Channel in the CNS of Biomphalaria glabrata Under Parasitic Infection
Authors: Vicente‐Rodriguez Laura Cristina , Méndez‐De Jesús Paola , Rosa‐Casillas Mariela , Hernández Anthony , Ramírez Cayra , Rosenthal Joshua , Miller Mark 
Published: June 2021

Schistosomiasis is a disease of major global concern for health and socioeconomic development. This neglected tropical disease represents a significant health burden for over 80 countries in Africa, Asia, and South America. Is estimated that more than 240 million of people required preventive treatment in 2016, and thousands are at risk of death annually. Schistosoma mansoni, the trematode specie that causes the most widespread form of intestinal schistosomiasis, requires two host organisms: snails from the genus Biomphalaria as its intermediary host, and humans or other mammals as its definitive host. Snails undergo major physiological and behavioral changes upon parasitic infection, but the neural contribution to these changes is poorly understood. As FMRFamide‐related neuropeptides regulate diverse physiological processes in the snail, such as respiration, reproduction and feeding, and these vital processes are known to be affected by parasitosis, we hypothesized that this peptide system participates in the behavioral and physiological modifications that occur in the snail following the parasitic infection. The FMRFamide peptides have two types of receptors, a G‐coupled receptor and a peptide gated sodium channel (FaNaC) that is unique to this phyla and is the sole sodium channel gated by a peptide. We performed immunohistochemistry and western blot experiments in snail CNS samples to determine the FaNaC receptor distribution and expression. We found a change in the neuronal distribution of the receptor in the visceral and left parietal ganglia from infected snails. Our overarching goal is to better understand the neural circuits involved in the physiological changes observed in B. glabrata during parasitic infection. This knowledge could lead to effective and selective strategies to control B. glabrata proliferation.Support or Funding InformationNIH (USA): RCMI MD007600, NIGMS‐RISE R25 GM061838NSF (USA): CREST HRD‐1137725, PIRE OISE 1545803 National Academy of Sciences (NAS, USA): U.S.‐Egypt Science and Technology (S&T) Joint Fund 2000007152; Science and Technology Development Fund (STDF, Egypt): USC17‐188This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of a Membrane‐Bound Insect Transferrin
Authors: Najera Diana G , Coca Michelle , Nutsch Kayla E , Gorman Maureen J 
Published: June 2021

Transferrins are extracellular proteins that bind iron. Vertebrate transferrins have well‐characterized roles in iron transport and immunity, but the functions of transferrins in most other animals are poorly understood. The goals of this study are to identify the functions of transferrin‐3 (Tsf3) from Drosophila melanogaster (fruit fly), and to determine whether Tsf3 is conserved in other species of insects. Our experimental approach has been to predict important features of the protein, analyze gene expression, perform RNAi‐mediated knockdown in cultured cells, and use phylogenetic analysis to identify Tsf3 orthologs. An analysis of the Tsf3 amino acid sequence predicts that Tsf3 is extracellular and anchored to the plasma membrane. Putative iron‐binding residues are present in the carboxyl‐lobe, but iron‐binding residues appear to be lacking in the amino‐lobe; therefore, we predict that only the carboxyl‐lobe binds iron. Tsf3 is expressed at a low level throughout development in many tissues, but it is very highly expressed in prepupal salivary glands. Cells of the prepupal salivary glands are exceptional because of their extensive DNA replication and because they synthesize large amounts of glue proteins that are used by the insect to attach itself to a solid surface prior to pupation. High expression of Tsf3 in the prepupal salivary glands suggests that Tsf3 may participate in the uptake of iron in these glands, possibly to be used as a cofactor for enzymes required for DNA synthesis, or perhaps to be used in the glue biosynthesis process. To test the hypothesis that Tsf3 is used for iron uptake by insect cells, we are in the process of evaluating the effect of RNAi‐mediated knockdown of Tsf3 on cellular iron content. Toward this goal, we verified that Tsf3 is expressed in cultured insect cells, synthesized dsRNA that targets Tsf3, and optimized a method to measure cellular iron content. Finally, we used a reciprocal BLAST method to search for possible orthologs of Tsf3 in insects from 12 different orders. We found that all 13 species analyzed have putative Tsf3 orthologs. Our findings demonstrate that Tsf3 is likely to be an extracellular, membrane‐bound, iron‐binding protein that may be involved in cellular iron uptake in diverse species of insects.Support or Funding InformationNSF IOS 1656388This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Uptake of Sialic Acids by Macrophage Cell Lines under Inflammatory Conditions
Authors: Zhao Yu , Sun Xue‐Long , Wang Dan 
Published: June 2021

Sialic acids (SAs) are the terminal sugars of cell surface glycoconjugates and are the critical determinants in many physiological and pathological processes. Macrophage cell surface expresses a dense layer of glycans often terminated with SAs which are involved in many biological processes of macrophages such as host‐pathogen recognition, migration, and antigen presentation. Notably, humans make Neu5Ac but are incapable of synthesizing Neu5Gc. However, humans can incorporate Neu5Gc into glycoconjugates through dietary acquisition. Neu5Gc‐containing glycoconjugates can be immunogenic and associated with many diseases and disease progressions such as cancer and cancer development. Moreover, the Neu5Gc‐containing glycoconjugates contribute to infectious diseases as they can serve as receptors for pathogens or toxin. Because the incorporation of Neu5Gc carries negative consequences for humans, we investigated if human macrophage cells may intake Neu5Gc under different condition and have some effects on its functions. Here, we utilized THP‐1 macrophages as a model to examine its Neu5Ac and Neu5Gc intake under the normal and inflammatory condition which was induced by lipopolysaccharide (LPS). LC‐MS/MS was used to quantify the total Neu5Ac and Neu5Gc changes, and cell surface SAs lectin bindings confirmed by flow cytometry analysis. Also, their phagocytosis capacity was evaluated upon Neu5Ac and Neu5Gc under inflammatory conditions. Overall, we found that THP‐1 macrophages has a different intake of Neu5Ac and Neu5Gc under different conditions and affect its phagocytosis as well.Support or Funding InformationThis work was supported by Faculty Research Development Fund and the research fund from the Center for Gene Regulation in Health and Disease (GRHD) at Cleveland State University supported by Ohio Department of Development (ODOD). The authors acknowledge the National Science Foundation Major Research Instrumentation Grants for supporting NMR and QTrap 5500 mass spectrometer instrument requisition at Cleveland State University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.