bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cardiac function in a chemical toxin (organophosphate) induced model of gulf war Illness: benefit of exercise training
Authors: Machi Jacqueline Freire , Conti Filipe , Schmidt Rodrigo , Morris Mariana 
Published: June 2021

BackgroundGulf War Illness (GWI) is a debilitating disease characterized by a cadre of neural, immunological and autonomic symptoms. We show that exposure to a sub‐lethal dose of Sarin lead to autonomic imbalance, cardiomyopathy and central nervous system changes. Diisopropyl fluorophosphate (DFP), an analogue of Sarin (GB), can be used as surrogate agent to produce neurotoxicity and neuro‐inflammation. There is limited information on the use of DFP to produce model of GWI with emphasis on the study of cardiovascular‐autonomic dysfunctions. Consideration was given to the inclusion of exercise training (ET) as part of the treatment of cardiac dysfunction induced by organophosphate exposure.ObjectiveThe aim was to determine the effect of chronic exercise training to the response to DFP on autonomic and cardiovascular function.MethodsMale C57BL/6J mice were divided into two groups (n=8/group): Sedentary (S) and trained (T) before and after DFP exposure. ET was performed on an electronic wheel at low intensity for 1 h/day, 5 days/wk. GWI model was induced by corticosterone (CORT) administered for 7 days (200 mg/l) followed by DFP (1.5 mg/kg, s.c). Cardiac function and ECG were assessed by transthoracic echocardiography (echo) under isoflurane anesthesia (2 – 2.5% on 0.8 l/min of O2).ResultsThe low dose of DFP produced left ventricular dilation on sedentary group, specifically, LV dilation of end systolic area (ESA) (S: 10.34±0.86 vs S+DFP: 6.69 ±0.41 cm2). The ET ameliorated DFP induced toxicity (T: 7.99 ±0.40 vs T+DFP 7.66±0.96cm2). Cardiac Output (CO) was reduced in sedentary group after DFP (S: 20.61±2.63 vs S+DFP: 14.78±0.85 ml/min), not change was observed in the ET group, revealing the efficacy of exercise protocol (T: 16.99±1.69 vs T+DFP: 17.90±1.42 ml/min). Myocardial performance index (MPI) and E/A ratio were significantly increased in S group after DFP exposure (MPI: S: 0.52±0.03 vs S+DFP: 0.60±0.02; E/A: S: 1.29±0.09 vs S+DFP: 1.64±0.07) changes that were not observed on T group (MPI: T: 0.50±0.04 vs T+DFP: 0.48±0.03; E/A: S: 1.35±0.09 vs S+DFP: 1.56±0.10) suggesting protection against DFP. Finally sympathetic modulation (LF) was increased in S group compared initial values and T group, as well as LF/HF ratio. Vagal modulation was similar between both groups.ConclusionIn conclusion, results demonstrate that DFP exerts a negative impact on cardiac/autonomic function. Our critical finding is that chronic exercise prevented most of the cardiovascular dysfunctions elicited by DFP, documenting the positive effects of exercise. This treatment protocol maybe equally applicable to other animal models or even humans.Support or Funding InformationThis work was funded under U.S. Department of Defense grant W81XWH‐13‐2‐0085This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Paralytic hypo‐energetic state facilitates anoxia tolerance despite ionic imbalance in adult Drosophila melanogaster
Authors: Campbell Jacob B. , Andersen Mads Kuhlmann , Overgaard Johannes , Harrison Jon F. 
Published: June 2021

Oxygen limitation plays a key role in many pathologies, and yet we still lack a fundamental understanding of the mechanisms responsible for variation in anoxia tolerance. Most vertebrate studies suggest that anoxia tolerance involves the ability to maintain cellular ATP despite the loss of aerobic metabolism. However, insects such as adult D. melanogaster are able to survive long periods of anoxia (LT50: ~8 h) in a hypo‐energetic state characterized by low [ATP]. In this study, we tested for possible mechanisms that allow D. melanogaster adults to survive long periods of anoxia. The adults are paralyzed within 30 sec and after two hours of anoxia, ATP was 3% of normal, [K+]o increased approximately threefold, pH dropped 1 unit, but survival was 100%. With 0.5–6 h of anoxia exposure the live adults maintained low but constant ATP levels while [K+]o and pH continued to change. When these animals were returned to normoxia they could restore hemolymph [K+]o and subsequent movement. However, with longer durations of anoxia, ATP levels decreased and [K+]o rose further, and both of these variables correlated tightly with decreased survival. This response contrasts with the anoxia‐sensitive larval stage (LT50: ~1 h). During anoxia larvae attempt escape for up to 30 min and after two hours of anoxia, ATP was <1% of resting, hemolymph [K+]o increased by 50%, hemolymph pH fell by 1 unit, and survival was zero. The superior anoxia tolerance of adult D. melanogaster appears to be due to the capacity to maintain a paralytic hypometabolic energy state with low but non‐zero ATP levels, and to be able to tolerate extreme extracellular ionic variability. This study suggests that a focus of research in anoxia‐tolerance should be the mechanisms by which animals can survive and quickly recover from such energetic and ionic conditions.Support or Funding InformationSupported by NSF IOS1256745.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Long‐term Delivery of “Low Dose” Repetitive Intermittent Hypoxia is Not Associated with Detectable Pathology
Authors: Gonzalez‐Rothi Elisa Janine , Allen Latoya A. , Santiago‐Moreno Juan , Ciesla Marissa C. , Asa Zachary A. , Smith Kristin N. , Tadjalli Arash , Perim Raphael , Santiago Juliet V. , Holland Ashley E. , Stefan Kelsey A. , Ross Ashley , Satriotomo Irawan , Kelly Mia N. , Simon Alec K. , Poirier Amy E. , Seven Yasin B. , Yarrow Joshua F. , Mitchell Gordon S. 
Published: June 2021

Repetitive acute intermittent hypoxia (rAIH) is a promising therapeutic strategy to induce spinal plasticity and improve respiratory and non‐respiratory function after chronic, incomplete spinal cord injury. Although brief rAIH exposures (< 2 weeks) have proven safe in rodent models and humans with cSCI, the safety profile of prolonged rAIH exposures is unknown. This is an essential step in the translation of rAIH to treat humans with respiratory and non‐respiratory paralysis due to injury and/or disease. Here we report an absence of detectable respiratory, cardiovascular, neurological, renal, liver and osteological pathologies after prolonged rAIH (3 months) in rats. Beginning 8 weeks post‐injury spinally intact rats, or rats with C2 hemisections (C2Hx) were treated 4 times per week for 3 months with: 1) rAIH (10, 5 min episodes of 10.5% O2; 5 min normoxic intervals) versus 2) sham normoxia. Whole body plethysmography was conducted following the exposures to assess respiratory function in spontaneously breathing rats. To assess cardiovascular pathology: 1) arterial blood pressure was assessed in urethane anesthetized rats, and 2) the heart and aorta were processed for histological analyses. To assess neurological pathology, the cortex, hippocampus and cervical spinal cord were harvested and histology was used to assess possible gliosis and/or cell death. The liver and kidney were processed for histological analyses, and all tissues are being evaluated by a board certified veterinary pathologist. Lastly changes in bone density were evaluated in the left and right humerus bones using microcomputed tomography. No differences in respiratory or cardiovascular function were observed following rAIH exposures (n = 8–12 per group; all P > 0.50), nor did we observe any evidence of pathology in the heart, aorta, liver or kidney. We found no evidence of hippocampal cell death and/or reactive gliosis based on cressyl violet, NeuN, cd11b and GFAP staining. Bone density analyses are pending, but preliminary results indicate significant bone loss only as a result of C2Hx, but not rAIH. These results add to a building body of evidence that prolonged, “low dose” rAIH is a safe, simple and effective means to improve respiratory and non‐respiratory motor function after chronic spinal cord injury.Support or Funding InformationDoD SC130298 CDMRP; NIH R01 HL69064 and OT2OD023854 (SPARC); NIH T32 HD043730 (LLA), NIH K12 HD055929 (EGR); and the McKnight Brain Institute.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating a novel regulation on a checkpoint protein Sda that is essential for biofilm formation and sporulation in Bacillus subtilis
Authors: He Yinghao , Gozzi Kevin , Qin Yuxuan , Chai Yunrong 
Published: June 2021

Bacteria have evolved various protection mechanisms, such as biofilm formation and sporulation, to help them adapt to and survive extreme environments. In the soil bacterium Bacillus subtilis, Sda is a “checkpoint” protein that plays a critical role in sporulation and biofilm formation by regulating the phosphotransfer from the histidine kinase KinA that culminates in phosphorylation of the transcription factor Spo0A, a master regulator for both sporulation and biofilm formation. Recent studies have shown a serine codon hierarchy‐based novel signaling mechanism for biofilm formation in B. subtilis. Interestingly, the sda gene has a much higher percentage of serine codons (12%) compared to the genome average (5%) in B. subtilis and a biased distribution of different serine codons. We hypothesize that the serine codon hierarchybased signaling mechanism also regulates Sda in that translation of Sda will slow down during nutrient limitation at the onset of biofilm formation and sporulation. To test our hypothesis, we constructed six Sda point mutants with synonymous substitutions in three TCN serine codons. We designed biofilm and sporulation competition assays to determine the impact of those synonymous changes in serine codons. We found that the Sda mutant with AGC substitutions outcompeted other mutants and became the dominant strain after several rounds of competition in both biofilm and sporulation assays. Our data indicates that synonymously switching serine codons in sda gene can delay biofilm formation and sporulation. This study suggests that this codon‐based serine sensing mechanism may be widely present in B. subtilis.Support or Funding InformationNational Science FoundationNortheastern UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying the Substrate Proteins of E3 Ubiquitin Ligase by Orthogonal Ubiquitin Transfer (OUT)
Authors: Yin Jun , Wang Yiyang , Bhuripanyo Karan , Chen Geng , Zhou Li , Liu Ruochuan , Zhou Han 
Published: June 2021

Ubiquitin (UB) is transferred through an E1‐E2‐E3 cascade to cellular proteins and encodes diverse signals such as proteasome degradation, subcellular trafficking, interacting with other proteins, and enzyme catalysis. The 2 E1s, 50 E2s, and more than 600 E3s constitute a complex network of UB transfer and make it a significant challenge to identify the substrates of a specific E3. Here, we developed a method that we called “orthogonal UB transfer (OUT)” to identify the direct ubiquitination targets of an E3. In this approach, an engineered UB (xUB) is exclusively transferred through an engineered cascade of xE1‐xE2‐xE3 to the substrates of a specific E3 (“x” designates engineered enzymes that are free of cross‐reactivities with their native partners). We used phage and yeast cell surface display to construct the OUT cascades of HECT E3 E6AP, U‐box E3s E4B and CHIP, and Ring‐between‐Ring (RBR) E3s HHARI and Parkin to profile their substrates. We found key signaling proteins such as kinases, phosphatases, and methyltransferases are ubiquitinated by these E3s. Based on the substrate profiles from the OUT screen, we discovered new pathways to regulate protein ubiquitination and degradation in the cell. Overall, we validated OUT as a powerful proteomic platform to identify the substrates of an E3 and map its position in the cell signaling networks.Support or Funding InformationThis work is supported by the National Institutes of Health (GM104498) and the National Science Foundation (1420193 and 1710460).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structural Determination of Inteins from Halophilic Organisms
Authors: Reidy Deirdre A. 
Published: June 2021

Protein splicing is a post‐translational process by which an intervening polypeptide, or intein, can excise itself and ligate the flanking polypeptides, or exteins. Two extreme halophilic archaea, Halobacterium salinarum (Hsa) and Haloquadratum walsbyi (Hwa), have proteins with inteins. Previous work in the lab demonstrated that splicing of the Hsa DNA Polymerase II (Polll) intein is salt‐dependent. In order to solve the structure of the intein, we have begun to screen for and optimize crystallization conditions. Structural studies will be done by x‐ray crystallography and/or high resolution NMR. We plan to do similar work with Hsa Cell Division Control Protein 21 (Cdc21) and Hwa Gyrase B (GyrB) to determine the structures of the inteins. Previous work demonstrated that the splicing of the Hsa Cdc21, like Hsa PolII, is salt‐dependent. In contrast, the Hwa GyrB intein is likely to have been a recent invader of the haloarchaea and has been shown to splice efficiently in E. coli without salt incubation. We are interested in determining the structure of these inteins from haloarchaea using the methods described.Support or Funding InformationThis work was supported by the National Science Foundation (grants MCB‐1244089 and MCB‐1517138 to KVM), the Dreyfus Foundation (KVM), and William F. McCall Jr. (DAR).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Homing Endonuclease and Protein Splicing Activity of Inteins from Extreme Thermophiles
Authors: Comeau Hannah Y. , Gomez Alvin V. , Mills Kenneth V. 
Published: June 2021

Protein splicing is a post‐translational modification by which intervening polypeptides, or inteins, catalyze their own excision from two flanking polypeptides, or exteins, concomitant with the ligation of the exteins. Many inteins contain a homing endonuclease domain, which can facilitate intein homing by catalyzing double strand breaks in intein‐less extein alleles. We studied the in vitro protein splicing and endonuclease activity of homing endonuclease‐containing inteins that interrupt the DNA Polymerase II (PolII) from the deep‐sea thermophiles Thermococcus kodakarensis (Tko) and Thermococcus barophilus (Tba). It was found that the Tba and Tko inteins can be purified as unspliced precursors and induced to splice in vitro at temperatures greater than 37°C. Additionally, when incubated at pressures up to 1500atm, the Tba and Tko inteins continue to exhibit splicing activity. Tba and Tko inteins also have temperature‐dependent endonuclease activity; the Tba intein is enzymatically active at pressures up to 1000atm. Future work will focus on identifying the active site residues of the homing endonuclease within the Tba and Tko inteins as well as determining the target sequence specificity of the enzyme. We also will determine if there is a difference in homing endonuclease activity between excised inteins and unsplicing intein precursors.Support or Funding InformationThis work was supported by the National Science Foundation (grant MCB‐1517138) and by the Camille & Henry Dreyfus 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.
Analyzing Protein Kinase C Domain Interactions with the Formin SepA
Authors: Olsen Elisabet , Betton Brianna , Atiq Zainab , Singh Ashmeet , Campbell Lynsey , Parish Paul C. , Hill Terry , Jackson‐Hayes Loretta 
Published: June 2021

This research focuses on understanding polarized growth in filamentous fungi. In the model organism Aspergillus nidulans and other filamentous fungi, polarized growth is focused at both the hyphal tip and septa, and growth at both areas involves deposition of cell wall material. To better understand polarized growth, we study proteins that localize to growing hyphal tips and forming septa. In A. nidulans, protein kinase C (PkcA) is involved in the organism's cell wall stress response, and our previous work uncovered a protein‐protein interaction between PkcA and the formin SepA. We also illustrated complementary roles of the two proteins in growth and cell wall integrity. Both PkcA and SepA localize to hyphal tips and septation sites. Using bimolecular fluorescence complementation, we found that SepA and PkcA physically interact at both sub‐cellular locations, and that this interaction occurs through amino acid residues within the C‐terminal half of SepA. Previously, we showed using a series of fluorescence‐labeled PkcA truncated proteins that two regions of PkcA are responsible for PkcA localizing to hyphal tips and septation sites. The first region is a 10 amino‐acid sequence near the carboxyl end of the C2 domain that is required for localization to hyphal tips and septation sites, while the second region is between the C2 and C1B domain, and is sufficient for localization to septation sites only. Here we present our progress towards determining which PkcA domains are involved in its interaction with SepA by using the previously published PkcA truncations in Gal4‐based yeast two hybrid assays. These results help to shed light on the mechanism underlying SepA's and PkcA's complementary involvement in polarized growth.Support or Funding InformationNational Science Foundation Grant 1615192 (Loretta Jackson‐Hayes, PI)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 Intersection of Learning Progressions and Concept Inventories: Assessment of Student Understanding of Acid/base Concepts
Authors: Wolfson Adele J. , Reed Charlotte R. , Mercer Allison M. , Sutheimer Susan , Lewis Jennifer E. 
Published: June 2021

Learning progressions are descriptions of students' growing sophistication in their understanding of a particular construct through a curricular sequence. Learning progressions are particularly useful for organizing complex constructs where students do not necessarily connect concepts as taught in different courses. One such complex construct is acid/base chemistry, which is a vital underpinning to much of biochemistry. We have developed a hypothetical learning progression for student comprehension of acid/base chemistry based on empirical studies. Learning progressions are only useful, however, if we can assess students' arrival at particular steps along the progression. To that end, we have compared the types of questions asked on concept inventories in chemistry, biology, and biochemistry, as well as common standardized tests. We find that none of these tests or surveys probes students' mastery of acid/base chemistry beyond the most elementary level. Therefore, there is a need for new assessment tools that target student understanding of the complex problems and relationships addressed in upper division chemistry and biochemistry courses.Support or Funding InformationSupported by an award from the National Science 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.
Mechanism of Membrane Biogenesis
Authors: Naik Akshata Ramesh , Kuhn Eric R. , Lewis Kenneth T. , Kokotovich Keith M. , Maddipati Krishna R. , Chen Xuequn , Hörber Heinrich , Taatjes Douglas J. , Potoff Jeffrey J. , Jena Bhanu P. 
Published: June 2021

Biological membrane in addition to delimiting the cell and its intracellular compartments, govern a wide range of life processes including ion transport, cell division, development, growth, and migration. The addition of new membrane to existing cellular membranes may involve fusion of preformed proteoliposomes and or the incorporation of intracellular free lipids and proteins. It is reported that supported lipid bilayers with adhered lipid vesicles when laterally stretched, expand due to fusion of the adhered vesicles. Mathematical modeling of the plasma membrane suggests trafficking of lipids into stretched membranes to relieve tension. Cell swelling has been reported during mitosis and mechanical forces are implicated in morphogenesis and development. However, the process of addition of new membrane to preexisting membranes in live cells is poorly understood. Here we report in live cells, that in addition to membrane stretch, composition of the existing membrane dictates membrane growth through the selective incorporation of both proteins and lipids.Support or Funding InformationWork presented in this article was supported in part by the National Science Foundation grants EB00303, CBET1066661 (BPJ); the WSU Interdisciplinary Biomedical Systems Fellow‐ship and Graduate Research Assistantship (ARN), the WSU Interdisciplinary Biomedical Systems Fellow‐ship and Graduate Research Assistantship (KTL). All authors critically analyzed results and proof read the manuscript. The authors declare no competing financial interests or conflicts.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human COQ10A and COQ10B are distinct putative StART domain proteins that restore Q biosynthesis and function in yeast
Authors: Tsui Hui S. , Pham Nguyen V. , Fernández‐Del‐Río Lucía , Amer Brendan R. , Bradley Michelle C. , Clubb Robert T. , Clarke Catherine F. 
Published: June 2021

Coenzyme Q (ubiquinone or Q) functions as an essential redox‐active lipid in respiratory electron and proton transport in cellular energy metabolism, and it is an important lipid‐soluble antioxidant in cellular membranes. In Saccharomyces cerevisiae, proteins Coq3‐Coq9, as well as Coq11, assemble into a multi‐subunit protein complex called the CoQ‐synthome, which is required for Q biosynthesis. Coq10, a putative steroidogenic acute regulatory (StAR)‐related lipid transfer (StART) domain protein is not a member of the CoQ‐synthome, but is required for the proper assembly of the CoQ‐synthome, efficient de novo Q biosynthesis during early‐log phase, and the function of Q in respiration and as an antioxidant. Humans possess two isoforms, namely COQ10A and COQ10B. Based on the RNA‐seq data from NCBI, COQ10A is predominantly expressed in heart while COQ10B is present in all tissues, suggesting that COQ10B is probably serving a more general role. Previous studies have shown rescue of S. cerevisiae coq10Δ respiration deficient phenotype by expression of human COQ10A. Here we present new evidence showing rescue of S. cerevisiae coq10Δ by expression of either the human COQ10A or COQ10B homolog, as determined by restoration of respiratory growth on non‐fermentable carbon sources, de novo Q biosynthesis, as well as restoration of the CoQ‐synthome.Support or Funding InformationThis research was supported by NSF MCB‐1330803 and Ruth L. Kirschstein National Research Service Award GM007185.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.