bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Novel Components in Fibroblast Growth Factor Signal Transduction
Authors: Webb Jason , Gaudenzi Carolina , Eichelberger Eric , Stefinko Mariya , Alonso Yanely , Ahmed Fardin , Voisine Cindy , Stern Michael , Lo Te‐Wen 
Published: June 2021

Fibroblast Growth Factor Receptors (FGFRs) are a type of receptor tyrosine kinase (RTK) that control many developmental processes. These include cell proliferation, cell migration, and cell survival. Mutations to these FGFRs and their pathways in humans result in developmental disorders such as achondroplasia, craniosynostosis, and thanatophoric dysplasia. To better understand FGFR signaling specificity we use the nematode C. elegans as a model. C. elegans contain a single FGFR, EGL‐15 which is imperative for sex myoblast migration and fluid homeostasis. Defects in the processes mediated by the EGL‐15 FGFR result in striking phenotypes that can be used to discover additional signaling components of the EGL‐15 pathway. For example, hyperactivation of EGL‐15 signaling causes the worm to accumulate excessive amounts of a clear fluid in its body which is known as the Clr phenotype. The isolation of suppressors of Clr (soc) mutants has helped to identify many of the core components of EGL‐15 signaling. A previous soc screen identified the adaptor protein SEM‐5 which links RTK activation to the activation of the RAS/MAPK pathway.SEM‐5 is known to bind to EGL‐15 at two tyrosine residues (Y1009 and Y1087). However, previous work suggests that removal of these binding sites does not confer a Soc phenotype. To verify this, we utilized three different CRISPR/Cas9 mutants. One mutant contains the Y1009F mutation, the second mutant contains the Y1087F mutation, and the third mutant contains both the Y1009F and Y1087F mutations. Each of these mutations was tested for their ability to suppress the Clr phenotype associated with the hyperactivation of the EGL‐15 signaling pathway. Neither the Y1009F or Y1087F mutation alone or together, results in a Soc (suppressor of Clr) phenotype. This would suggest that a key component that links EGL‐15 to SEM‐5 has yet to been identified. We performed an additional modified suppressor screen to isolate these potential “links” between EGL‐15 and SEM‐5 and are in the process of mapping and identifying these components.Support or Funding InformationIthaca College H&S Summer Scholars, Ithaca College CSTEP and FERP Funding, NIH R15 (RGM122001A), MNEIU Student Center For Science EngagementThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of UCP2 protein expression in normal RIN‐5F and UCP2 inhibited cells
Authors: Willis Jacquelyn , Clemons Tameka 
Published: June 2021

Uncoupling protein 2 (UCP2) is a mitochondria integral membrane protein that transports protons across the intermembrane space. Traditionally, the function of UCP2 is to impact oxidative phosphorylation by uncoupling ATP synthesis. However, a cell signaling role for UCP2 has recently been investigated. UCP2 expression has been shown to have expression in pancreatic beta cells and the presence of UCP2 has been suggested to lead to a decrease in the release of insulin. Although amylin is co‐secreted with insulin, studies on the relationship between amylin and UCP2 have had little investigation. A known function of amylin is to block glucagon secretion, which leads to a reduction in the rate of glucose production by decreasing the rate of gastric emptying. This study analyzed the protein expression level of amylin in normal RIN‐5F cells and RIN‐5F cells that have been transfected with a UCP2 siRNA construct. The study included stimulating the cells with either 11mM or 30 mM of glucose and comparing the protein expression level in the two cell lines. Western blot analysis was conducted and the results suggest a difference in UCP2 and amylin levels in normal RIN‐5F cells in comparison to the transfected RIN‐5F cells.Support or Funding InformationNational Science Foundation Research Initiation Award #24430This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Cushing Syndrome Mutation of Protein Kinase A C‐subunit Disrupts the Internal Allosteric Network Affecting Regulation and Substrate Specificity
Authors: Walker Caitlin , Wang Yingjie , Olivieri Cristina , Karamafrooz Adak , Casby Jordan , Bathon Kerstin , Taylor Susan S. , Calebiro Davide , Bernlohr David A. , Veglia Gianluigi 
Published: June 2021

Genetic alterations of the PRKACA gene coding for the catalytic subunit of the CAMP‐dependent protein kinase A (PKA‐C) are linked to cortisol‐secreting adrenocortical adenomas resulting in the onset of Cushing's syndrome. Among those, a single mutation (L205R) located in the P+1 loop of the kinase's active site has been found in 30–60% of patients. It has been proposed that L205R hinders the interactions of PKA‐C with its regulatory (R) subunit generating a constitutively active kinase causing hyperphosphorylation of downstream substrates. However, the x‐ray structures of PKA‐C wild‐type and L205R are essentially identical and the aberrant mechanism of function of the mutant remains unknown. To gain mechanistic insights into the multifarious effects of this single mutation, we carried out solution NMR spectroscopy in concert with binding thermodynamics as well as molecular dynamics simulations. We found that this single mutation, in addition to changing the protein‐protein interface, disrupts the internal allosteric network of the kinase and the network of interactions between the small and large lobes. The ternary complex of L205R with nucleotide and pseudo‐substrate displays significantly altered allosteric interactions, leading to the disruption of the interactions with R‐subunit, the binding with endogenous protein kinase inhibitor (PKI), and modifying substrate specificity. Our results show that this single mutation has global effects on the structure of the kinase that leads to the dysregulation and aberrant phosphorylation profiles.Support or Funding InformationThis work was supported by the National Institutes of Health, GM100310 (G.V.) and S10 OD021536 (G.V.)., and the CRC/Trasregio 166 (Project C1 to DC) as well as the IZKF Würzburg (grant B‐281 to DC). KB was partially supported by a fellowship through a grant of the German Excellence Initiative to the Graduate School of Life Sciences, University of Würzburg.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TRPM8 channel role in mesenchymal stem cells osteogenic differentiation
Authors: del Pilar Torres Yolima , Henao Juan Carlos , Barreto Alfonso , Rodríguez Viviana , Mejía Camila 
Published: June 2021

Mesenchymal stem cells (MSCs) have properties that make them a good model to use in regenerative medicine. MSCs proliferation and differentiation are complex processes involving the activation of several molecules that induces changes in intracellular calcium concentration. Different ion channels are responsible for calcium influx including TRP channels. The present study evaluates the expression and functional activity of TRPM8 channels in human mesenchymal stem cells. Cells were characterized by the expression of the specific surface markers CD90+, CD105+ and CD34− and by the ability to differentiate into adipogenic and osteogenic linage. Western blot, immunocytochemistry and flow cytometry were performed to determine TRPM8 protein expression. Patch clamp recordings and fluorescence imaging with Fluo‐4 AM were used to analyze channel activity. We found that the agonist menthol induces an increase in ionic currents which were incompletely abolished by treatment of the cells with the antagonist BCTC. Changes in intracellular calcium concentration were detected after addition of menthol, suggesting an effect induced by activation of TRPM8 channels. Alizarin red staining showed that the TRPM8 agonist Icilin increased osteogenic differentiation. On the contrary, the TRPM8 antagonist BCTC decreased cell differentiation. These results were confirmed by real time PCR with the marker Alkaline Phosphatase (ALP). Results from this study suggest that the activity of TRPM8 channels in mesenchymal stem cells is related with regulation of osteogenic differentiation.Support or Funding InformationThis work was supported by Departamento Administrativo de Ciencia, Tecnología e Innovación, COLCIENCIAS, Project: Evaluación del papel de canales TRPM8 en la diferenciación de células madre mesenquimales derivadas de tejido adiposo; Grant ID 120365843092.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interactions between doripenem and NDM metallo‐β‐lactamases as inspiration for future generation antibiotics
Authors: Orischak Matthew , Shurina Ben , VanPelt Jamie , Page Richard 
Published: June 2021

Antibiotic resistant bacterial infections are a growing threat to patients globally. Bacteria are evolving rapidly and developing new modes of antibiotic resistance. One of these resistance modes is the expression of a family of proteins that chemically inactivate antibiotics. β‐lactamases act on antibiotics that contain a β‐lactam ring such as penicillin. New Delhi metallo‐β‐lactamase (NDM) was selected as the primary focus for this project as it confers multi‐drug resistance, is an increasing threat to public health, and there are currently no FDA approved inhibitors for NDM. Doripenem is an antibiotic notable for its relative stability as a carbapenem, a subclass of β‐lactam antibiotics, and is often used as a last resort antibiotic. Unfortunately, NDM is capable of hydrolyzing doripenem. Despite this, doripenem may still serve as a promising scaffold from which new derivatives could be designed for use as next generation antibiotics that are resistant to β‐lactamase mediated hydrolysis. Alternatively, doripenem derivatives might be designed to act as inhibitors for β‐lactamases such as NDM. In order to better engineer doripenem derivatives for either of these purposes, structural information detailing the interactions between NDM and doripenem is required. While there are existing structures of NDM in complex with other substrates such as meropenem and imipenem, there was no structure of doripenem in complex with NDM. Our lab has successfully solved an X‐ray crystal structure of hydrolyzed doripenem bound to the active site of NDM‐4 at 2 Å resolution. Our crystal structure of NDM‐4 in complex with doripenem reveals novel interactions between the sulfonamide functional group at the R2 position of doripenem and the surface of NDM‐4. Previously published crystal structures of NDM complexed with carbapenem substrates do not show strong interactions with the functional groups at the R2 position. Future work will explore doripenem derivatives and investigate their resistance to hydrolysis or potential use as β‐lactamase inhibitors.Support or Funding InformationNIH R35 GM128595This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Isoleucine Serves as a Precursor for the Synthesis of β‐Alanine in Plants
Authors: Rouhier Kerry A , Perrett Morgan E , Goldfarb Margo H , Reinhart Peter E , Wilczewski‐Shirai Kai C , Raymond‐Schmidt Lisa , Gothard Megan E , Piras Crystal A 
Published: June 2021

The non‐protein amino acid β‐alanine serves as an athletic performance enhancer and as a key precursor in the production of food additives and pharmaceuticals. In plants, β‐alanine is important for the synthesis of pantothenate (vitamin B5) and subsequently coenzyme A through a condensation reaction with pantoate. It is well established that the source of pantoate is valine, however the synthesis of β‐alanine in plants is less‐well established. There is some evidence that plants synthesize β‐alanine from spermine (or other poly‐amines), uracil, and propionate, but evidence has only been recorded in very specific plant systems, such as stress‐tolerant Limonium latifolium. In this project we sought to confirm the synthesis pathway of β‐alanine in Arabidopsis and the more economically relevant grain, wheat. We hypothesized that another branched‐chain amino acid, isoleucine, could serve as a source of β‐alanine in plants. Isoleucine degradation concludes with the production of propionyl‐CoA, which is further metabolized to acetyl‐CoA in a similar manner to β‐oxidation. Therefore, in order for isoleucine to serve as a precursor, the intermediate malonate semialdehyde must react via an unknown aminotransferase to produce β‐alanine. Using transgenic Arabidopsis plants and Hard Red Winter wheat, seedlings were treated with isotopically‐labeled precursors and extracts were analyzed by both NMR and GC‐MS. Results show that β‐alanine can be synthesized from both propionate and isoleucine via the proposed propionyl‐CoA metabolic pathway. Given these new findings and the prior evidence that valine serves as the precursor for pantoate, branched‐chain amino acids look to play an important role in the synthesis of pantothenate and coenzyme A in plants.Support or Funding InformationClare Boothe Luce Undergraduate Scholars program funded by the Henry Luce Foundation (Grant #9601), Kenyon Summer Science Scholars Program funded by Kenyon College, Kenyon Summer Science Cascade Program funded by the Sherman Fairchild FoundationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Increase of Efficiency of RuBisCO
Authors: Link Tina , Stanovski Lisa , Fisher Joseph , Kona Abhishek , Lu Allison , Nguyen Lan , Rajput Aashita , Saberi Zara 
Published: June 2021

Carbon dioxide (CO2) is the primary source of carbon for life on Earth. As such, the efficient incorporation of CO2 into organic matter is of significant importance. Ribulose‐1,5 bisphosphate carboxylase/oxygenase (RuBisCO), sequenced from the cyanobacteria Synechococcus PCC6301, is a 64.8 k‐Da photosynthetic enzyme involved in the oxidation‐reduction process of carbon fixation, assimilating inorganic carbon into bioavailable organic carbon. RuBisCO is composed of two subunits: a large L chain and a smaller S chain, with a length of 467 amino acids. Each chain consists of eight sub‐chains and weighs 51.87 k‐Da and 12.93 k‐Da respectively. RuBisCO, the most abundant protein in the world, is slow‐reacting, fixing about 3–10 CO2 molecules each second per molecule of enzyme. During the catalysis of RuBisCO, the formation of an active site intermediate is able to react with either CO2 or Oxygen (O2). Approximately 25% of the time, RuBisCO will bind with O2 instead of CO2. This misbinding has a significant impact on the enzyme's effectiveness within the Calvin cycle as well as other photosynthetic processes. The functionality error of RuBisCO means that the enzyme exhausts large quantities of ATP in order to process the O2 and then attempt to bind to CO2. Minimizing RuBisCO's intermediate active site's availability can reduce the occurrence of photorespiration. While variations to RuBisCO exist between different photosynthetic species, all species experience inefficiencies due to oxygenation to some extent. Efforts to discover more efficient carboxylation through random mutagenesis of RuBisCO lead to the discovery of a mutant that catalyzes the carbon fixation reaction with greater speed, accuracy, and precision. The resultant mutated RuBisCO was chosen through selection in E. coli. Mutations in the C‐terminal loop 6 region and a substitution mutation at 331 of valine to alanine, both within the large subunit, decreased the specificity factor by almost 40%. RuBisCO's enlarged specificity factor could potentially benefit the development and growth of many different species of plants. The potential increase in crop productivity could have a substantial impact on global food distribution. Walton High School SMART Team has designed a 3D model of RuBisCO using JMOL to investigate the relationship between structure and function.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Avocado Oil on Metabolic Profile and Development of Diabetic Nephropathy in Goto‐Kakizaki Rats
Authors: Ortiz‐Avila Omar , Saavedra‐Molina Alfredo , Cortés‐Rojo Christian 
Published: June 2021

Uncontrolled glucose and lipid metabolism during diabetes, caused by a decrease in the synthesis of regulatory substances such as insulin and adiponectin, promotes the development of diabetic complications. One of the most frequent and devastating complications is diabetic nephropathy, which is characterized by high levels of proteinuria due to inadequate glomerular filtration. The aim of this study was to evaluate the effect of avocado oil on metabolic profile and the development of diabetic nephropathy in a model of type 2 diabetes.Goto‐Kakizaki rats were used as a model of type 2 diabetes, and avocado oil was administered orally for 3, 6 and 12 months. The results showed that treatment with avocado oil maintained for a longer time adiponectin levels, however had no effect on insulin levels. The treatment improved the lipid profile in rats by lowering cholesterol, triglyceride and LDL levels and maintaining HDL concentrations. Another important aspect was that avocado oil did not result in weight gain in diabetic rats. When evaluated renal function, the oil partially counteracted the increase in proteinuria observed in the diabetic group, while in the histopathological evaluation diabetic rats showed signs of diabetic nephropathy such as accumulation of extracellular matrix in the glomerulus and thickening of the basement membrane starting from 3 months of treatment. Treatment with avocado oil delayed the development of these alterations until 6 months.These results show that avocado oil improved several parameters of the metabolic profile and delayed the development of diabetic nephropathy, probably due to the components on avocado oil, since some of them have antioxidants functions, while others, such as oleic acid, have a beneficial effect on the metabolism of lipids and carbohydrates.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Effects of De Novo Lipogenesis Inhibitors on plasma lipids and liver PNPLA3
Authors: pocai alessandro 
Published: June 2021

Obesity and genetic polymorphisms such as the patatin‐like phospholipase domain‐containing 3 (PNPLA3‐I148M; 49% allele frequency in Hispanics) predispose to non‐alcoholic steatohepatitis (NASH). NASH patients are at increased risk of death by cardiovascular and liver‐related complications. Hence, there is an urgent need to develop new strategies for preventing and treating NASH. Increased de novo lipogenesis (DNL) and liver lipid deposition are processes associated with the early stage of the disease.Inhibition of liver acetyl‐CoA carboxylase (ACCi) in NASH patients reduces DNL and liver triglycerides (TG) and improves noninvasive markers of fibrosis. Unexpectedly, ACCi results in a significant rise in plasma TG, a major concern given the already high risk of CV disease in NASH patients. Hence, alternative strategies to reduce DNL devoid of the lipid liability might be beneficial for liver steatosis in NASH. Previous studies have demonstrated that diacylglycerol acyltransferase 2 inhibition (DGAT2i) lowers DNL and improves inflammation and fibrosis in rodents. Here, we conducted a 2‐week study in diet‐induced obese (DIO) mice comparing the effects of ACCi versus DGAT2i on lipid metabolism.Thirty‐six DIO mice were treated with vehicle, ACCi (30 mg/kg PO) or DGAT2i (10 mg/kg and 30 mg/kg PO). ACCi and DGAT2i resulted in significant reduction in liver fat mass (−43% for ACCi vs. −23 and −29% for DGAT2i), however, ACCi significantly increased plasma TG (+49%, ACCi treatment vs. vehicle; p < 0.001). ACCi increased mRNA expression of liver sterol regulatory element‐binding protein 1c (SREBP‐1c, +40%) and of the lipogenic genes stearoyl‐CoA desaturase‐1 (SCD1; +650%), ELOVL fatty acid elongase 6 (ELOVL6; +373%), and Glycerol‐3‐phosphate acyltransferase 1 (GPAT1; +96.4 %). In the same study, DGAT2i resulted in no change of plasma TG while reducing expression of SCD1 (−92% and −95%), ELOVL6 (−16% and −10%) and GPAT1 (−32% and −39%) mRNA. DGAT2i decreased PNPLA3 expression (−71% and −79%; DGAT2i 10 and 30 mg/kg vs. vehicle; p<0.05) while ACCi increased PNPLA3 (+2400%, ACCi vs. vehicle; p<0.001). PCSK9 plasma levels were significantly reduced (−67%; p < 0.001) in DGAT2i treated mice. In contrast, ACCi treated mice exhibited a significant increase of plasma PCSK9 levels (226%; p < 0.001).Inhibition of de novo lipogenesis with DGAT2 and ACC inhibitors are potential promising strategies to lower liver lipids in NASH patients. Our results also demonstrated that DGAT2i and ACCi inhibition have opposite effects on liver PNPLA3 and plasma PCSK9This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Functional decline in brown adipose progenitors during BAT remodeling
Authors: Ruan Hai‐Bin , Huang Zan , Zhang Zengdi 
Published: June 2021

In humans, there is a striking decrease in the prevalence and activity of brown adipose tissue (BAT) with age and obesity; however, the cellular and molecular mechanisms of BAT remodeling are almost completely unknown. Rodent models have been widely used to study BAT but they possess constitutively active BAT throughout life even at thermoneutrality; therefore, complementary animal models that possess characteristics of human BAT remodeling are needed. Here, we found that BAT progressively whitened in rabbits in a similar manner as in humans. As rabbits mature, preadipocytes in BAT displayed reduced capacity to be differentiated into brown adipocytes, when transplanted into immune‐deficient mice. RNA‐sequencing of brown preadipocytes identified that, during BAT remodeling, genes enriched for muscle contraction and mitochondrial function were down‐regulated, while genes involved in lipid and amino acid metabolism were up‐regulated. Knocking down genes regulating muscle and mitochondrial function in rabbit brown preadipocytes blunted their ability to become mature brown adipocytes in vitro and in vivo. We also identified follistatin like 1 (FSTL1) as a novel “batokine” that is specifically expressed by brown preadipocytes but not mature brown adipocytes. FSTL1 gradually reduced its expression when BAT whitens in rabbits, but was stably expressed in mouse BAT during life. Deleting FSTL1 specifically in the Myf5 lineage in mice reduced BAT size, UCP1 expression, and body temperature in newborns, suggesting the constitutive expression of FSTL1 prevents BAT whitening in mice. Taken together, we identify potential molecular mechanisms underlying functional decline in the browning ability of preadipocytes that contribute to BAT remodeling in rabbits and possibly humans.Support or Funding InformationAmerican Diabetes Association (18‐IBS‐167)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.