bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Rising of Acylsugar Diversity: Metabolic Innovation in Tomato Trichomes through BAHD Enzyme Promiscuity and Pathway Evolution
Authors: Fan Pengxiang , Miller Abigail M. , Jones A. Daniel , Liu Xiaoxiao , Last Robert L. 
Published: June 2021

In cultivated (Solanum lycopersicum) and wild tomatoes, glandular trichomes produce structurally diverse acylsugars. These insect protective metabolites are mixtures of aliphatic esters of sucrose and glucose, with acylchains varying in length from C2 to C11. We are analyzing acylsugar biosynthesis in the cultivated tomato, along with the biochemical and evolutionary mechanisms that generate acylsugar diversity across wild tomatoes (1, 2). Previous studies identified four BAHD family Acylsucrose Acyltransferase (ASAT) enzymes. These catalyze consecutive reactions from sucrose and acyl‐CoA substrates to produce the full set of cultivated tomato acylsucroses in vitro (3). Recently we identified a modified ‘upside down’ acylsugar pathway in the wild tomatoes S. pennellii and S. habrochaites that produces unusual types of acylsucroses, namely triacylsucroses with all three chains on the pyranose ring (P‐type) (4). This is in contrast with the furanose ring acylated acylsucroses (F‐type) produced in cultivated tomatoes and a variety of wild tomato species. Biochemical analysis demonstrates that the acyl acceptor substrate specificities were changed in two S. pennellii ASAT orthologous enzymes, which reversed their enzymatic activity order, leading to the ‘flipped pathway’. A phylogeny‐based structure‐function analysis of the ASAT2 and ASAT3 enzymes led to identification of a small number of amino acid changes that switch their activities between the F‐ and P‐ acylsucrose pathways (4, 5). Investigation of these pathways in other tomato species allowed inference of the evolutionary events giving rise to divergence of F‐ and P‐type acylsucroses across tomato species. This work demonstrates multiple evolutionary mechanisms in biochemical pathway evolution, including gene duplication and loss, amino acid substitution, and how the emergence of enzyme promiscuity restructured a specialized metabolic pathway and led to metabolic product innovation.Support or Funding InformationThis work was funded by National Science Foundation grants IOS‐1025636 and IOS‐PGRP‐1546617 to A.D.J. and R.L.L. Abigail Miller was supported by NSF REU grant DBI‐1358474 in the summer of 2014 and an American Society of Plant Biologists Summer Undergraduate Research Award during the summer of 2015. A.D.J. acknowledges support from the USDA National Institute of Food and Agriculture, Hatch project MICL‐02143.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lipidomic Profiling Reveals Soluble Epoxide Hydrolase As a Therapeutic Target of Obesity‐induced Colonic Inflammation
Authors: Wang Weicang , Yang Jun , Zhang Jianan , Wang Yuxin , Hwang Sung Hee , Wan Debin , Sanidad Katherine Z. , Yang Haixia , Liu Jun‐Yan , Park Yeonhwa , Liu Zhenhua , Hammock Bruce D. , Zhang Guodong 
Published: June 2021

Obesity is associated with enhanced colonic inflammation, which is a major risk factor of colorectal cancer. Considering the obesity epidemic in United States, there is an urgent need to identify novel therapeutic targets for obesity‐induced colonic inflammation. Eicosanoids are endogenous lipid signaling molecules involved in regulating inflammation and immune responses. Using a LC‐MS/MS‐based lipidomics approach, we found that obesity‐induced colonic inflammation is associated with increased expression of soluble epoxide hydrolase (sEH) and its eicosanoid metabolites, termed fatty acid diols, in colon tissue. To study the functional role of sEH, we treated mice with sEH inhibitors, TPPU or t‐TUCB, in high fat diet (HFD)‐induce obesity model, and found that pharmacological inhibition of sEH attenuated obesity‐induced colonic inflammation. Administration of sEH inhibitors diminished obesity‐induced infiltration of immune cells and expressions of pro‐inflammatory cytokines in colon tissue. Furthermore, we examined the sEH−/− mice in obesity model, and found genetic ablation of sEH reduced colonic concentrations of fatty acid diols and decreased the expressions of pro‐inflammatory cytokines. Finally, HFD treatment increased expression of phosphorylated GSK3β in colon tissue, while such effect was attenuated in sEH−/− mice, or by treatment with sEH inhibitors, suggesting that pharmacological inhibition and genetic ablation of sEH attenuated HFD‐induced activation of Wnt signaling in colon. Together, these results support that sEH could be a novel therapeutic target for obesity‐induced colonic inflammation and associated diseases.Support or Funding InformationThis work was supported by USDA NIFA grants 2016‐67017‐24423 and 2014‐67017‐21762, NIEHS grant R01 ES002710 and NIEHS Superfund Research Program P42 ES004699, and National Natural Science Foundation of China (NSFC) grants 81702832, 81672938 and 81470588.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CYTOCHROME P450 EPOXYGENASE EICOSANOID PATHWAY AS NOVEL BIOMARKER AND THERAPEUTIC TARGET OF COLORECTAL CANCER
Authors: Zhang Guodong , Wang Weicang , Yang Jun , Edin Matthew , Hammock Bruce , Zeldin Darryl 
Published: June 2021

In US, there are ~130,000 new cases and ~50,000 deaths resulted from colorectal cancer (CRC) every year, making CRC the second leading cause of cancer‐related deaths. There is an urgent need to identify novel biomarkers and/or therapeutic targets of CRC, in order to reduce the risks of CRC. Here we report that the expressions of cytochrome P450 (CYP) epoxygenases and their eicosanoid metabolites (fatty acid epoxides) are dramatically increased in cell culture model, animal model, and human cancer patients of CRC; in addition, pharmacological inhibition or genetic ablation of CYP epoxygenases suppresses development of CRC in mice, illustrating that the previously unappreciated CYP epoxygenase pathway could serve as novel biomarkers and/or therapeutic targets of CRC. In a well‐established azoxymethane (AOM)/dextran sulfate sodium (DSS)‐induced CRC model in C57BL/6 mice, LC‐MS/MS showed that CYP‐derived fatty acid epoxides were the only eicosanoid metabolites which were significantly increased in both plasma and colon tissues of tumor‐bearing mice. RT‐PCR and immunoblotting analysis demonstrated that the expressions of CYP (including CYP2C38, 2C39, 2C50, 2C65, 2C70, and 2J6) were significantly increased in colon tumors. Consistent with the animal results, LC‐MS/MS showed that the serum levels of CYP‐derived fatty acid epoxides were significantly increased in patients clinically diagnosed with CRC. To study the functional roles of CYP in CRC, we tested whether pharmacological inhibition or genetic ablation of CYP affects CRC development in mice. We found that treatment with SKF‐525A and clotrimazole, which are two different inhibitors of CYP epoxygenases, suppressed AOM/DSS‐induced CRC in mice. We further showed that compared with wild‐type (WT) control mice, the AOM/DSS‐induced CRC was reduced in Cyp2c gene cluster knockout mice. To elucidate the specific metabolites involved in the pro‐CRC effects of CYP, we treated mice with epoxyoctadecenoic acids (EpOMEs) which are metabolites of linoleic acid produced by CYP, and found that EpOMEs promoted development of colonic inflammation in mice. Together, our studies showed that the previously unappreciated CYP epoxygenase pathway plays critical roles in promoting progression of CRC, in part through formation of pro‐inflammatory EpOMEs. These results suggest that the circulating levels of CYP‐derived fatty acid epoxides could serve as novel diagnostic biomarkers of CRC, and the CYP enzymes could be promising therapeutic targets of CRC.Support or Funding InformationThis work was supported by USDA NIFA 2016‐67017‐24423 (to G.Z.). In addition, the research is supported in part by NIEHS R01 ES002710 and Superfund Research Program P42 ES04699.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ibuprofen increases resistance to exertional heat stroke in female mice
Authors: Garcia Christian Kyle , Alzahrani Jamal , Mattingly Alex , Laitano Orlando , Robinson Gerard , Murray Kevin , Clanton Thomas 
Published: June 2021

The impact of nonsteroidal anti‐inflammatory drugs (NSAIDs) on the severity or susceptibility to exertional heat stroke (EHS) is unknown. Previous studies have demonstrated that in humans, thermogenic responses to endurance exercise are suppressed by NSAIDs and that NSAIDs increase the endurance time to exhaustion in rats. However, NSAIDs have also been shown to contribute to organ damage in both severe exercise alone and in conditions of heat stress alone, which may limit performance.METHODSIn this study, we determined the effects of ibuprofen administration (IBU) on exercise performance and resistance to heat stroke in male and female mice in an established model of exertional heat stroke (EHS). Two days prior to EHS, the mice were given continuous IBU‐loaded chow (ad libitum) at 375 mg/kg (N = 7–8) compared to controls (N =44). To induce EHS, the mice ran a preprogrammed incremental exercise protocol using a forced running wheel in an environmental chamber at 37.5°C/35% RH, until a symptom‐limited max was reached (loss of consciousness). The EHS protocol was defined by an incremental exercise phase (IP) and a steadystate exercise phase (SP). During the IP, wheel speeds were increased every 10 min until core temperature (Tc) = 41°C. This was followed by a SP phase, in which speed was maintained until a symptom‐limited endpoint.RESULTSAll groups ran to nearly identical peak Tcs (42.2–42.4°C). IBU‐treated female mice exercised longer in heat, prior to collapse, compared to controls (223 vs. 176 min; P = 0.0018), but there was no significant effect in males (133 min vs 124 min). Time in the IP was higher in the IBU‐female mice compared to controls (181 vs 131; P=0.0018). Though the duration of the SP phase did not change in any group, female IBU mice ran at significantly higher power outputs during the SP phase compared to controls (females: 34.5 mW vs 27.4 mW, P=0.0004) demonstrating a lower rate of heat accumulation for a given rate of external power expended.CONCLUSIONSThese data demonstrate that in female mice, short‐term IBU treatment improves exercise performance in the heat and delays the onset of EHS. The results are consistent with the hypothesis that a component of the temperature elevation seen in EHS arises from endogenous pyrogen production that can be blocked by IBU. The lack of similar responsiveness in males may reflect greater sensitivity to EHS in male vs. female mice and predominance of other mechanisms limiting performance in the heat. Authors' views not official US Army or DoD policy. W81XWH‐15‐2‐0038Support or Funding InformationAuthors' views not official US Army or DoD policy. Department of Defense Grant W81XWH‐15‐ 2‐0038This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Thermoregulation Deficiencies in Mice with a Deletion in the Muscle Protein Titin
Authors: Miyano Carissa Ashley , Buck Charles Loren , Nishikawa Kiisa 
Published: June 2021

The muscular dystrophy with myositis (mdm) mouse carries a deletion in the muscle protein titin. A previous study found that mdm mice shiver at a lower than expected frequency for their body size, are mostly heterothermic with a narrow thermoneutral zone beginning at 34C, and have reduced active muscle stiffness in vivo compared to their wildtype siblings. Impairment in heat production (shivering thermogenesis) could be due to the N2A deletion in the titin protein leading to a more compliant muscle and a lower shivering frequency. The ability of mdm mice to use nonshivering thermogenesis via Uncoupling Protein 1 in brown adipose tissue to generate heat during cold stress may also be impaired and contributing to their heterothermy. The purpose of this study was to evaluate mechanisms for the inability of mdm mice to maintain homeothermy during cold stress and to further understand their metabolic properties. We hypothesized the inability of mdm mice to maintain homeothermy is due to an impairment in heat production (e.g. shivering thermogenesis and nonshivering thermogenesis) and that metabolic rate will reach a maximum at a higher ambient temperature than in wildtype mice. In a temperature experiment, mice were placed in a mouse cage that used indirect calorimetry to measure metabolic rate at four different temperatures (34C, 29C, 24C, 19C). Nonshivering thermogenesis was stimulated by administering 1.2 mg kg−1 of norepinephrine subcutaneously. In the temperature experiment, there was a significant interaction between genotype and temperature, with significant differences in metabolic rates at the lowest temperatures of 19C and 24C (Two‐Way ANOVA and Tukey HSD, p<0.05. Mdm mice had significantly lower body temperatures than wildtype mice at 29C, 24C, and 19C, and reached their maximum metabolic rate at 24C, suggesting that a temperature between 19–24C is the minimum at which mdm mice are able to sustain thermogenesis. Area under the curve (AUC) using the trapezoidal rule was used to calculate total thermogenic capacity following norepinephrine injection. Mdm mice had a lower overall AUC (Welch's T‐Test, p=0.03) as well as a lower overall time that they increased metabolic rate after norepinephrine injection (Welch's T‐Test, p=0.02). Our results suggest that in addition to a deficiency in shivering frequency, mdm mice also have a reduced capacity for nonshivering thermogenesis. The deficiency in shivering thermogenesis and nonshivering thermogenesis combined likely lead to inability to maintain euthermic body temperatures below 34C.Support or Funding InformationNSF IOS‐0742483, NSF IOS‐1025806, and NSF IOS‐1456868, the W. M. Keck Foundation, and the Technology Research Initiative Fund of Northern Arizona University.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lower Diffusing Capacity of the Lungs for Carbon Monoxide in Women with a Patent Foramen Ovale
Authors: Lovering Andrew , Schallerer Annalisa , Olson Travis , Davis James 
Published: June 2021

The test of diffusing capacity of the lungs for carbon monoxide (DLco) assumes that all of the cardiac output flows through the pulmonary circuit. Right to left blood flow across a patent foramen ovale (PFO) allows blood to bypass the pulmonary circuit. The shunted red blood cells are not exposed to alveolar gas and thus, would not bind any carbon monoxide with a DLco test. This may result in a lower carbon monoxide uptake and therefore a lower measured DLco. Consequently, we hypothesized that subjects with a PFO (PFO+) would have a lower DLco than subjects without a PFO (PFO−). We retrospectively analyzed historical data from our lab that included 77 PFO+ subjects (Ages:18–35; 31 Female) and 55 PFO− subjects (Ages 18–38; 24 Female). DLco was measured using the single‐breath, breath‐hold technique and ATS/ERS standards. We compared DLco absolute, DLco% predicted and DLco/Alveolar volume (VA) absolute and DLco/VA %predicted. There were no significant differences between PFO+ and PFO− subjects as a group. There were no significant differences between PFO+ and PFOmen. However, PFO+ women had DLco and DLco/VA % predicted values that were 10% lower than PFO− women, though these values were still >100% predicted. Median and mean degree of resting PFO shunt scored with transthoracic saline contrast echocardiography were identical between men and women. In conclusion, these data suggest the degree of blood flow through the PFO under resting conditions may not explain the reason for the lower DLco observed in PFO+ women.Support or Funding InformationDepartment of Defense Grant W81XWH‐10‐2‐0114American Heart Association Scientist Development grant 2280238American Physiological Society's Giles F. Filley Memorial Award for Excellence in Respiratory Physiology and MedicineAmerican Lung Association in Oregon and administered through American Thoracic Society Grant C‐10‐014This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Multisubunit Multiactive Site DNA Polymerase Complexes with Coordinated Activities
Authors: Trakselis Michael A. , Cranford Matthew , Chu Aurea M. 
Published: June 2021

Duplication of the genome requires efficient, rapid, and coordinated activities of many enzymes that interact within a larger replisome complex. However, whether those interactions are stable or stochastic is currently being tested. These enzymatic abilities include high processivities, fidelities and plasticities to ensure faithful genome maintenance in spite of many obstacles to DNA synthesis. Many multisubunit enzyme subcomplexes have been identified within the replisome and most stabilize enzyme complexes on DNA for continued and processive enzymatic action over long stretches of the genome. One of the most well characterized subcomplexes is the elongation complex minimally consisting of the DNA polymerase holoenzyme which includes accessory proteins known to stabilize the association of the polymerase and direct repeated catalysis.In particular, the stability of the DNA polymerase holoenzyme is sensitive to stable secondary DNA structures, protein/DNA complexes, and various DNA damage. High fidelity (HiFi) DNA polymerases in complex with their accessory proteins are responsible for the bulk of genome synthesis, however, most organisms encode multiple separate specialized DNA polymerases to aid in overcoming genomic obstacles with translesion synthesis (TLS). The presence of anywhere from 2 to 18 separate DNA polymerases (depending on the organism) all with identical specificities for a 3′‐OH primer/template DNA creates a complex multiequilibrium within a cell. Moreover, many of these DNA polymerases have been shown to directly or indirectly interact to enhance catalysis. For example, a HiFi trimeric DNA polymerase complex has increased kinetics and processivity explained through dynamic active site switching and exchange. Other direct DNA polymerase interactions allow for coordinated synthesis across DNA lesions through a concerted switch of active sites. Oligomeric DNA polymerase active site coordination will be shown using a variety of presteady‐state kinetic and thermodynamic assays. These and other DNA polymerase interactions and equilibria properties will be put into context and compared across different domains of life to illustrate DNA synthesis mechanisms that maintain a high degree of fidelity and plasticity in spite of a multitude of genomic obstacles.Support or Funding InformationBaylor UniversityAmerican Cancer Society (RSG‐11‐049‐01‐DMC)National Science Foundation (NSF1613534)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Epigenetic Regulations of Genes Related to Lipid Metabolism by MicroRNA in Mice Fed High Fat Diet
Authors: Tang Xinyu , Xu Guanying , Pan Yuan‐Xiang , Chen Hong 
Published: June 2021

BackgroundHigh fat diet impacts lipid metabolism by altering the transportation, oxidation, and storage of fatty acids. Lipoprotein lipase (LPL) plays a critical role in lipid metabolism by catalyzing triglyceride hydrolysis and lipoprotein uptake in multiple tissues. Previous study reported that miR‐29b negatively regulated LPL expression in mammary epithelial cells transfected with miR‐29b mimics. The present study investigated changes in LPL expression and epigenetic mechanisms by miR‐29 family in different tissues in mice fed a high fat diet.MethodFive‐week old male CBA mice were fed with either a control diet (Con group, 10% kcal from diet) or a high fat diet (HF group, 45% kcal from diet) ad libitum for 11 weeks.ResultsLPL mRNA was increased in adipose, muscle and colon in response to high fat diet. However, LPL mRNA expression decreased in the liver by high fat diet as well as hepatic lipase (HL). The results also showed the highest expression level of LPL mRNA in adipose tissue, followed by muscle, colon, and liver. Cpt1a mRNA, an important gene related to beta oxidation, was increased in adipose, muscle, colon, and liver in the HF group. Moreover, Cpt1a mRNA expression was the highest in liver followed by adipose, muscle, and colon. Meanwhile, high fat diet reduced the expression of miR‐29a, a predicted suppressor of LPL from miR‐29 family, in adipose tissue.ConclusionThe present results indicated that the triglyceride hydrolysis could be activated by high fat diet in other tissues than the liver and fatty acid transportation could be induced in mitochondria. Overall, LPL and Cpt1a mRNA changed differently in multiple tissue in response to high fat diet and is potentially post‐transcriptionally regulated by microRNA.Support or Funding InformationSource of research support (in a separate textbox): Campus Research Board of University of Illinois at Urbana‐Champaign and the USDA Cooperative State Research, Education and Extension Service, Hatch project numbers # ILLU‐698‐923 and ILLU‐698‐379This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.