bluuebunny/crossref_metadata_2025_split huggingface dataset:
Skeletal Muscle Metabolic Enzyme Activity at Varying Temperatures in the Thirteen‐lined Ground Squirrel (Ictidomys tridecemlineatus)
Authors: Conley Emily , Symanietz Sarah , Loeza‐Rodriguez Monserrat , Schlater Amber E 
Published: June 2021

Hibernation is a prolonged season where some mammals experience a dramatic reduction in food intake while undergoing a series of torpor bouts for weeks or months at a time. During individual torpor bouts, metabolic rate decreases and hibernators will thermoconform to their environment, dropping their core body temperature (TB) close to ambient temperature (TA). The causality of the reduction of metabolic rate has yet to be fully understood. Thirteen lined ground squirrels, Ictidomys tridecemlineatus, endure a hibernation season of about 8 months. Although, the thirteen‐lined ground squirrels undergo hibernation for a long period of time, they still manage to preserve and maintain their organs without any greater damage and are able to achieve physiological sustainability. Given the extreme range of TB experienced throughout the hibernation period, which is unlike most other mammalian endotherms (that typically reside TB at a constant 37°C), in addition to the unique metabolic challenges of thermoregulation within and between torpor bouts, we explored function of metabolic enzymes and subsequent substrate utilization across a range of physiologic temperatures in skeletal muscle tissue. We performed functional enzyme assays of lactate dehydrogenase (LDH, marker of anaerobic metabolism) and citrate synthase (CS, marker of aerobic metabolism) in the gastrocnemius muscle from control, non‐hibernating squirrels. Preliminary LDH data suggest that anaerobic activity is higher at 37°C as compared to thermoconforming temperature ranges of 30°C and 25°C. Alternatively, preliminary CS data suggest that aerobic activity does not change across a range of physiologic temperatures. Collectively, these aerobic and anaerobic comparisons across three different physiologic temperatures suggest that squirrels may not be suppressing aerobic metabolism when thermoconforming to lower temperatures, despite overall metabolic rate suppression during torpor bouts. Future studies may look to characterizing enzyme activity at an even lower range of physiologic temperatures, and may look to include data from hibernating squirrels in addition to non‐hibernating squirrels. These kinds of preliminary baseline data can ultimately be used to better understand the physiologic basis governing reduction of metabolic rate and subsequent substrate utilization during prolonged torpor bouts.Support or Funding InformationMcNair Scholars Program & The College of Saint Scholastica.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In Vitro and In Vivo Anti‐Hyperglycemic and Antioxidant Effects of Phenolic Compounds isolated from Oryza sativa L.(Keun‐nun‐Jami) Variety
Authors: Kim Tae‐Yang , Kang Hanna , Lee Jung‐Yun , Yu Jin‐a , Hong Sung‐Gil , Kwon Young‐In 
Published: June 2021

Type 2 diabetes, one of the chronic diseases, is a problem in both developed and developing countries. In particular, excessive ingestion of grain, such as rice, can result in hyperglycemia and type 2 diabetes. Therefore, a Oryza sativa L. variety, Keun‐nun‐Jami (KNJM) extracts were investigated for their anti‐hyperglycemic effect using in vitro and animal models. In vitro study inhibitory activities of both ethyl alcohol extracts of bran and germ against rat intestinal sucrase and maltase were evaluated. Oxygen radical absorbance capacity (ORAC) was also investigated. Ethyl alcohol extract of KNJM bran (EKB) had the highest sucrase inhibitory activity followed by ethyl alcohol extract of KNJM germ (EKG). Taxifolin, a flavonol, was isolated from the extract of KNJM along with three unknown compounds using HPLC and semi‐prep HPLC systems. Their structures were elucidated by extensive spectroscopic analysis, including 1D and 2D NMR, and FTICR‐MS. Furthermore, post‐prandial blood anti‐hyperglycemic effect of the extacts and taxifolin were measured using SD (Sprague Dawley) rat model. Postprandial blood glucose levels were tested for 3 hr after sucrose and starch administration with or without 500 mg of EKB, EKG, and taxifolin, respectively. Similar to result in vitro study, EKB which has higher content of taxifolin than that of EKG had stronger anti‐hyperglycemic effect than that of EKG. These results indicate that taxifolin from KNJM serves to reduce post‐prandial blood glucose rise and slower absorption of glucose in the small intestine, via carbohydrate hydrolyzing enzymes inhibition. These findings suggest that EKB has better potential for this bioactivity and this observation possibly correlates with the taxifolin contents observed in both extracts. Selected variety of colored rice, KNJM may provide the potential to contribute as a dietary strategy for controlling postprandial hyperglycemia.Support or Funding InformationThis work was supported by a grant from the Next‐Generation BioGreen 21 Program (No.PJ01314003), Rural Development Administration, Republic of KoreaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The relationship between food exposure frequency and expression of AMP‐activated protein kinase (AMPK) mRNA in the brain, liver, and muscle of channel catfish
Authors: Kostner Danica LeAnn , Dougherty Megan , Schmidtberger Abigail , Kobayashi Yass 
Published: June 2021

Obesity poses serious challenges to our healthcare system with serious consequences of associated diseases. Our laboratory has been examining how the expression of genes involved in food intake and tissue nutrient metabolism is associated with the development of obese‐like phenotypes in channel catfish. The objective of the study was to examine changes in expression of AMPK mRNA in response to changes in feeding frequency. Fish were fed every 12 (overfed), 24 (control), or 48 (underfed) hours (n= 4 tanks per treatment, 8 fish per tank) for 28 days. Brain, liver, and muscle tissue were collected at the end of the study. The expression of the AMPK mRNA was examined using quantitative RT‐PCR. The muscle expression of all AMPK subunit mRNA was unaffected by feeding frequency. In contrast, brain expression of alpha 1 and beta 1 subunit mRNA had a linear tendency for increase in response to increased feeding frequency. Hepatic expression of alpha 2 and beta 1 mRNA was lower in the overfeed group compared to those assigned as control or underfeed. Hepatic expression of alpha 1 mRNA decreased linearly in response to increased feeding frequency. Our results demonstrated that the hepatic and the brain expression of AMPK mRNA was influenced by increase in exposure to food in a transcript‐specific manner. Given that the AMPK serves as a regulator of tissue nutrient sensing and metabolism in mammals, it is possible that AMPK may share a similar function in the brain and liver but not in the muscle of channel catfish.Support or Funding InformationKansas IDeA Network of Biomedical Research Excellence Network (NIH/NIGMS grant number P20GM103418)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bisphenol A, but not Bisphenol S, exposure increases lipid deposition by acting on the PI3K pathway in vivo
Authors: Gibert Yann , YOGANANTHARAJAH Prusothman , McGee Sean , Meikle Peter , Mellett Natalie , Basheer Faiza , Hall Liam 
Published: June 2021

Bisphenol A (BPA) is an endocrine disrupter present in many daily products including food and beverage packaging. BPA exposure has been linked to many disorders in humans including cancer, neurological disorders, diabetes and obesity and has therefore been declared toxic and a partial ban is in effect in several countries. Recently the plastic industry has come with a bisphenol analog as a substitute for BPA in BPA free products: BPS. However to date, little is known regarding the toxicity of BPS in vivo. In this study, we used the zebrafish as a vertebrate model to understand the mechanism of action of BPA and BPS in lipid usage during embryogenesis. Zebrafish embryos exposed to the extreme low dose 1 nM BPA (over 6,000‐fold lower than the accepted human daily exposure), have a 65% increase in lipid deposition. Interestingly, BPS exposure, at the same concentration, has no effect on lipid deposition. BPA is classified as an estrogenic compound however, pharmacological blockage of the nuclear estrogen receptors does not suppress the induced lipid deposition. To identify which pathway might be involved in this increased lipid deposition, we performed a full embryo lipidomic analysis. This revealed that BPA exposed embryos had a dramatic increase of diacylglycerol (DAG) and Phosphatidylinositol (PI) lipids. Strikingly, lipidomic analysis of BPS exposed embryos showed a decrease of DAGs and PIs. This is the first study to demonstrate a phenotypical difference between BPA and BPS at similar doses. Double exposure of BPA and LY294002, a strong inhibitor of phosphoinositide 3‐kinases, abolishes the BPA induced increase in lipid deposition and lipid usage. At the molecular level, BPA exposure increases the expression of key enzymes of the PI3K pathway such as: cdipt, plcb3, vps34p and p110. Our study allows us to propose a model in which BPA, but not BPS, at very low concentration can activate the PI3K pathway leading to an accumulation of lipid during embryogenesis in an estrogen receptors independent manner. Our results confirm that, at least in terms a of lipid usages, BPS is safer than BPA exposure and support the ban on BPA from consumer merchandise.Support or Funding InformationThis project was supported by an Australian Diabetes Research Trust (DART) research grant to YGThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phosphorylation Dependent Interactions of Perilipin 5 with Adipocyte Triacylglycerol Lipase
Authors: Osborn Rachel E , Tansey John T 
Published: June 2021

Diseases such as non‐alcoholic fatty liver disease, obesity and type II diabetes are on the rise in the US, and the dysfunction of cellular lipid metabolism is a major contributor to them. Perilipins are a class of five proteins found on the exterior of lipid droplets which play a role in proper lipid metabolism. Perilipin 5 is expressed in tissues that are oxidative such as the liver, heart and skeletal muscle. We used CHO cells expressing perilipin 5 protein and immunofluorescence microscopy to examine lipase interactions with perilipin 5 under different phosphorylation states. We hypothesized that the interactions of lipases such as adipocyte triacylglyerol lipase (ATGL) and perilipin 5 were phosphorylation dependent. Using immunofluorescence microscopy, we confirmed that perilipin 5 changes its subcellular localization and moves from lipid droplets and cytosol to the nucleus when stimulated with forskolin and IBMX to activate the PKA signaling pathway. Identification of lipase‐perilipin 5 interactions are ongoing. Collectively these data show that perilipin 5 interactions and localization in the cell are phosphorylation dependent.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lipid Droplet Protein Binding in Response to Altered Phospholipid Composition
Authors: Renier Timothy , Paetz Olivia , Vuong Sunny , Listenberger Laura 
Published: June 2021

Alcoholic Fatty Liver Disease (AFLD) is the alcohol‐induced accumulation of excess lipids in hepatocyte lipid droplets. Lipid droplet surfaces are phospholipid monolayers predominantly composed of phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Protein binding to lipid droplets affects droplet size and activity. Previous experiments with a rat model of AFLD show lipid droplet surfaces have a decreased ratio of PC:PE. We aimed to determine if the altered phospholipid composition of AFLD lipid droplets leads to differential binding of several proteins of interest, selected for their membrane binding motifs and roles in lipid droplet homeostasis. By culturing AML12 cells in choline‐deficient media with oleate, an unsaturated fatty acid, we simulated the altered phospholipid composition of AFLD lipid droplets. Next, we isolated lipid droplet fractions and examined proteins of interest by SDS‐PAGE and western blotting. We found equivalent levels of rab18 and lanosterol synthase associating with lipid droplet fractions from cells grown with and without choline. Alternatively, there was an increase in the level of αCCT1 and perilipin 2 in lipid droplet fractions from cells grown without choline. By evaluating how altered phospholipid monolayer composition affects the binding of lipid droplet proteins, we seek to further elucidate the biochemical pathways that contribute to AFLD.Support or Funding InformationThis work was supported by St. Olaf College Collaborative Undergraduate Research and Inquiry, Howard Hughes Medical Institute, the St. Olaf TRIO McNair Scholars Program, and LSAMP Alliance.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis of Phosphorylation Sites in Perilipin 5
Authors: Sumser Natalie M , Tansey John T 
Published: June 2021

In modern times, there has been a significant increase in metabolic disease. Obesity has been a growing concern in the U.S, leading to a rise in various health problems, such as type II diabetes and cardiovascular disease. One major factor in these diseases is lipid metabolism. The understanding of this process would lead to a better understanding of these diseases. Perilipins, a family of five proteins associated with lipid droplets, play an important role in lipid metabolism. Perilipin 5 is primarily seen in oxidative tissue, acting to regulate fatty acid storage and release, as well as fatty acid oxidation. It is known that phosphorylation partially regulates the activity of Perilipin 5, particularly due to PKA activation, and we hypothesized that there are multiple phosphorylation sites on the protein. Treatment of cells with forskolin and IBMX was used to increase cyclic AMP to activate PKA. This was followed by immunopreciptation and trypsin digestion of perilipin 5. Phosphopeptides are enriched through affinity chromatography with a Fe‐NTA column prior to conducting mass spectrometry on the samples to differentiate between phosphorylation sites. We hypothesize that both a combination of where perilipin 5 is phosphorylated and when the sites are phosphorylated cooperate to regulate the function of the protein.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction of Perilipin 5 with Mitochondria
Authors: Todd Claire I , Tansey John T 
Published: June 2021

Problems with lipid metabolism can lead to array of diseases such as fatty liver disease, obesity, and diabetes. Nearly one‐third of the world's population is obese or overweight, and struggle with these diseases. Cellular lipids are found in lipid storage droplets. Perilipins are a family of proteins that associate with the surface of lipid storage droplets and regulate their contents. We used CHO cells expressing perilipin 5 protein and assayed them using immunofluorescent microscopy to determine if mitochondria interact with perilipin 5 under different phosphorylation states. We hypothesized that the interaction of mitochondria and perilipin 5 was phosphorylation dependent. We used antibodies that detect either FLAG tagged and native perilipin 5 and Mitotracker red to stain cells and forskolin and IBMX to activate the PKA signaling pathway. Following one hour of stimulation, perilipin 5 appeared in the lipid droplets and cytosol of the cell but interactions with mitochondria were not apparent. In this model 3 hours of stimulation is required for perilipin 5 to translocate to the nucleus. Collectively these ongoing studies show that the cellular partitioning of perilipin 5 is in part phosphorylation dependent.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The chinchilla as a novel animal model of gestational diabetes
Authors: Duvald Christina Søndergaard , Overgaard Maria Dahl , Ovesen Per Glud , Jeppesen Per Bendix , Pedersen Michael 
Published: June 2021

BackgroundGestational diabetes occurs in 3–4% of all pregnancies, thereby being one of the leading pregnancy diseases in developed countries. The disease poses important health implications for both mother and child. Several animal models exist in this field, but with significant limitations. Consequently we want to introduce the pregnant chinchilla as a novel animal model of gestational diabetes. The chinchilla is excelled for various reasons; it gives birth to one or two cubs, is pregnant for 115 day and has a histological similar placenta barrier as compared with the human one. Since the chinchilla has never been used in diabetic studies, we aimed to show that it is possible to make it diabetic, before making a gestational model.Streptozotocin is by far the most applied method to induce diabetes in animal models, mimicking the ethology of diabetes type 1. However, since gestational diabetes is due to insulin resistance, we wanted to use high fructose feed as a method to induce diabetes.MethodsTwelve chinchillas were included in the study, six of which received high fructose diet to induce diabetes and six who received control diets. Blood glucose was monitored using Abbott Libre Freestyle glucometer. After 19 weeks the animals were euthanized and organs and blood withdrawn for analysis.Preliminary resultsNon‐fasting plasma glucose levels in chinchillas fed with high fructose diet was 19.3 ± 6.9 mmol/l, compared to controls 11.1 ± 2.2 mmol/l, p=0.056. Furthermore, DXA scans showed significant higher body fat percentage among fructose animals, p=0.043 compared to controls. HBA1C was, however, not affected in any of the groups.ConclusionWe have proven it possible to induce high blood glucose levels in the chinchilla by using high fructose diet. Because of the low sample size the blood glucose levels were, however, not statistically significant between groups. In spite of this, we consider the chinchilla a promising option for a future model of gestational diabetes.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antioxidant and Haematological Activities of Ethanolic Extract of Ricinus communis
Authors: Ihekuna Ogechi , Imaga Ngozi Awa , Adewusi Banke 
Published: June 2021

Ricinus communis commonly known as castor oil is used as a traditional natural remedy or folkloric herb for the control and treatment of a wide range of diseases around the globe. Various studies have revealed the presence of diverse phytochemicals such as alkaloids, flavonoids, terpenes, saponins, phenolic compounds such as kaempferol, gallic acid, ricin, rutin, lupeol, and ricinoleic acid. The present study investigates the in vivo and in vitro analyses of the effect of ethanolic extract of ricinus communis leaf and seed on animal models. The antioxidant potential of Ethanolic leaf and seed extract of Ricinus communis was investigated. The result shows that the ethanolic extract of the seed and leaf exhibited IC50 values of 77.17± 0.36 and 69.33 ± 0.26 μg/mL, 62.86±0.54 and 62.39±0.18 μg/mL respectively in DPPH and nitric oxide radical inhibition assays these values were less than those obtained for ascorbic acid used as standard. In the in vivo experiments the extract treatment caused a significant increase in the level of catalase in the liver and the kidneys. A significant increase in the level of SOD in the liver was observed. The acute toxicity study was carried out in both male and female wistar rats; the results reveal that even at the highest dose 2000mg/kg body weight R.communis seed and leaf extract showed no significant toxic effects showing that LD50 is above 2000 and belongs to class 5. The haematological and biochemical parameters of the treatment groups were compared with the control group, result shows that there were no significant differences (mean ± SEM n=6 *p < 0.05) in the biochemical and haematological parameters of treatment groups compared to the control group. These findings suggest that the ethanolic extract of Ricinus communis seed and leaf exhibited a strong antioxidant activity which may be responsible for some of its reported pharmacological activities and can be used as antioxidant supplement.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comprehensive snapshots of an unusual reaction cycle for an atypical protein tyrosine phosphatases (PTP)
Authors: Wang Huanchen , Jessen Henning J. , Shears Stephen B 
Published: June 2021

Inositol pyrophosphates (PP‐InsPs) are “energetic” intracellular signals that are ubiquitous in animals, plants, and fungi. We are characterizing new proteins that regulate PP‐InsP metabolism and/or function as their ‘receptors'. Here, we describe our structural characterization of At1g05000, an Arabidopsis member of the cysteine‐based protein tyrosine phosphatase family that exhibits higher reactivity towards PP‐InsPs than phosphoproteins. We provide 20 high resolution crystal structures which describe how this atypical PTP has been repurposed to become the most active of known PP‐IPs phosphatases. These structures include complexes with different ligands including inositol pyrophosphates, polyphosphates and nucleotides; together they reveal pre‐reactant, reactant, intermediate and product states. As a consequence, we describe an atypical mechanism: an exceptionally electropositive ‘P’‐loop that immobilizes the β‐phosphate for hydrolysis; the opposite end of the substrate is anchored by Lys and Arg residues in a loop that is pushed into the binding pocket due to intramolecular crowding; a canonical catalytic acid is sacrificed to avoid electrostatic clashing with the substrate; a conserved water is identified to function as the proton donor; two catalytic important residues, His156 and Arg188, form π‐cation interactions that are important for positioning substrate and the catalytic water; hydrolyzed phosphate ion is rotated during catalysis. Overall, our data support a substrate‐driven reaction mechanism. Furthermore, this work opens up new directions for studying regulatory interactions of PP‐InsPs with human protein tyrosine phosphatases.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.