bluuebunny/crossref_metadata_2025_split huggingface dataset:
PKA RIα Holoenzyme Crystal Structure Reveals Its Allosteric Regulation and Carney Complex Disease Implications
Authors: Lu Tsan‐Wen , Wu Jian , Ahuja Lalima , Aoto Phillip , Sun Nicholas , Taylor Susan 
Published: June 2021

Protein Kinase A (PKA) or cAMP‐dependent protein kinase played an important role in numerous signaling pathway and significant biological processes, such as apoptosis, proliferation, cell cycles, mitochondria hemostasis, gluconeogenesis, etc. PKA has emerged the central role of cAMP‐dependent pathway.In the canonical cAMP‐dependent pathway, PKA activity is tightly controlled by cAMP. In the absence of cAMP, PKA is in inhibited holoenzyme form and composed by two regulatory subunits (R subunits) and two catalytic subunits (C subunits). When PKA holoenzyme (R2C2) binds with cAMP, R and C subunits dissociate and generate active form of free catalytic subunit and dimer of regulatory subunits.There are four PKA R subunits isoforms (RIα, RIb, RIIα, and RIIb) in human, and they are all functional non‐redundant. RIα and RIIα express ubiquitously, yet RIb and RIIb are more tissue specific. Although all four PKA R subunits share similar sequence and domain construction, they have different biochemical properties, cAMP sensitivities, and localization. In mice model, RIb knockout mouse show memory defect and RIIb knockout mouse have lean phenotype. However, RIα is the only R isoform that shows embryonic lethality in mice knockout model.Moreover, most of PKA‐related disease mutations are in RIα. Carney Complex (CNC) disease makes PKA activity increased by enhancing PKA RIα holoenzyme sensitivity toward cAMP. Acrodysostosis (ACRDYS) patients show hormone resistant and lower PKA activity.Here, we have shown RIα holoenzyme crystal structure which can reveal its allosteric regulation and mechanism. The holoenzyme structure can reflect its physiological state in vivo and show how RIα being regulated by ATP and affected by oxidative potential. The isoform‐specific interface can also explain how CNC disease mutations cause PKA dysfunction. All these indicate PKA RIα playing as a ‘master regulator’ of cAMP‐dependent signaling pathway in vivo.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Student vs Faculty Assessment on a Cadaver Handoff and its Correlation to Practical Grades
Authors: Elzie Carrie , Glen Dirk , Krimsier Jonathan , Van Winkle Katherine 
Published: June 2021

Assessment has become an integral part of development in medical education. Previous reports have shown that higher performing students are generally more critical of themselves, while lower performing students are the opposite. We decided to test this hypothesis in a first year medical school gross anatomy course using a “cadaver handoff” that mimics the patient handoff presentation in clinical settings. Specifically, we were interested in student's ability to self‐evaluate their own performances on the previous day's dissection presented to their peers. Our specific aims were to 1) determine if there are any recognizable differences between student self‐evaluations and faculty evaluations with regard to handoff presentations, 2) determine the impact of cadaver handoffs on overall student performance on practical exams, and 3) determine if feedback could improve student performance overtime.The EVMS gross anatomy course has 152 students of which half attend lab on Tuesdays and the other half attend lab on Thursdays. To improve continuity and communication skills, the “handoff” was created between the two groups. Prior to the presentation, presenting students completed a self‐evaluation on preparedness, confidence, dissection, anatomical knowledge, presentation skills and dissection journal. At the conclusion of the presentation, faculty filled out a similar rubric. Student versus faculty rubrics were compared to one another in each of those categories to address whether students were overconfident, realistic or more critical of their performance compared to faculty.Preliminary data collected from four labs shows no statistical difference between student evaluations and faculty evaluations; however, 10% of students had inflated performances, while another 10% had under‐rated performances. Further analyses of the remaining labs will be presented, as well as, stratification of data by overall student performance in the course and on practical exams. The vast majority (98%) of students thought performing the handoff better prepared them for the exam. Further data will be reported to support or negate this by comparing presentation scores to performance of relative practical exam questions.Overall, implementing a cadaver handoff in the medical gross anatomy course provides an opportunity for early exposure of students to presentation skills, self‐evaluations, feedback and reflection. These could have overall positive effects on exam performance.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MxiN Differentially Regulates Monomeric and Oligomeric Species of the Shigella Type Three Secretion System ATPase Spa47
Authors: Case Heather , Dickenson Nick 
Published: June 2021

Shigella rely entirely on the action of a single type three secretion system (T3SS) to support cellular invasion of colonic epithelial cells and to circumvent host immune responses. The ATPase Spa47 resides at the base of the Shigella needle‐like type three secretion apparatus (T3SA), supporting protein secretion through the apparatus and providing a likely means for native virulence regulation by Shigella and a much needed target for non‐antibiotic therapeutics to treat Shigella infections. Here, we show that MxiN is a differential regulator of Spa47 and that its regulatory impact is determined by the oligomeric state of the Spa47 ATPase, with which it interacts. Both in vitro and in vivo characterization find that MxiN interaction with Spa47 requires the six N‐terminal residues of Spa47 that are also necessary for stable Spa47 oligomer formation and activation. This interaction with MxiN negatively influences the activity of Spa47 oligomers while upregulating the ATPase activity of monomeric Spa47. Detailed kinetic analyses of monomeric and oligomeric Spa47 in the presence and absence of MxiN uncover additional mechanistic insights into the regulation of Spa47 by MxiN, suggesting that the MxiN/Spa47 species resulting from interaction with monomeric and oligomeric Spa47 are functionally distinct and that both could be involved in Shigella T3SS regulation. Uncovering Spa47 regulation by MxiN addresses an important gap in the current understanding of how Shigella controls T3SA activity and provides the first description of differential T3SS ATPase regulation by a native T3SS protein.Support or Funding InformationNational Institute of Health (1R15AI124108‐01A1)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Computational Histopathological Analysis of Nanoparticle Distribution in Breast Cancer Models
Authors: Henderson Elizabeth , Simons Brian , Korangath Preethi , Stewart Jacqueline , Ivkov Robert 
Published: June 2021

While many current nanoparticle studies focus on the macroscopic distribution of nanoparticles (e.g. pharmacokinetics applications), this study investigates the microscopic distribution of iron based nanoparticles within tumors through a combined histological and computational approach. Using breast cancer as a model (due to its wide variety of cell lines and well characterized animal models), we evaluated the distribution of antibody‐conjugated nanoparticles to identify and characterize their distribution patterns in xenografts with varying amounts of target protein expression.Five breast cancer cell lines (MDA‐MB‐231, MCF7/NEO, MCF7/HER, BT474, and HCC1954) with varying expression of HER2, were grown in Nude and NOD Scid Gamma (NSG) mice. When the tumor volume reached 150mm^3, five animals were assigned to one of three groups to receive intravenous injections of PBS, bio‐ionized ferrite (BNF)‐Plain or BNF‐HER nanoparticles (5mg/animal). 24 hours later, tumors were harvested and bisected. One half was fixed in 10% formalin, cross‐sectioned for histological Prussian blue analysis, and digitized. ImageJ was employed to visualize and characterize nanoparticle distribution patterns in the Prussian blue stained tumor cross‐sections from the five xenograft models referenced above. The tumor boundary was defined by a user‐drawn region of interest (ROI) and then divided into bands of equal areas. Each quantile was analyzed for pixel density and other spatial distribution statistics.We observed little evidence of BNF‐Plain uptake, but BNF‐HER was internalized about 3 times higher in all tumors, regardless of their HER2 status. For all animals that received BNF‐HER particles, each xenograft model showed marked differences in nanoparticle uptake and distribution. Between the different xenografts, HCC1954 showed the highest accumulation of nanoparticles, followed by BT474, MCF7/HER, MCF/NEO, and MDA‐MB‐231. Certain xenograft models also displayed differences in the heterogeneity of nanoparticle dispersion: nanoparticles in BT474 were spread across the tumor, while nanoparticles in HCC1954 tumors were tightly clustered at the tumor periphery. Despite the differences in uptake and heterogeneity, the highest concentration of nanoparticles was found closer to the tumor edge than the tumor center across all xenografts.Support or Funding InformationNIH, The Jayne Koskinas Ted Giovanis Foundation for Health and PolicyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the Biosafety of Petroleum Ether oil of Piper guineenses used as grain protectants using Wistar rats
Authors: Adeniyi Paul Ayomitunde , Gboyega Hakeem Ishola 
Published: June 2021

ObjectivesThe study was carried outto determine the biosafety of reported insecticidal potency of Piper guineenses petroleum extract (PPPE) used on stored products using wistar rats as a model.MethodologyPiper guineenses seeds were extracted using stardard procedures and the filtrate later concentrated using the rotary evaporator to obtain P.guineenses petroleum ether extract (PPPE). Twenty four rats were divided into four groups labeled I, II, III and IV (six rats per group). Group I served as the control group and were administered orally with Dimethyl Sulphoxide (DMSO) and normal saline, Group II were administered orally with 500mg/kg of PPPE, Group III were administered orally with 1000mg/kg of PPPE while Group IV were administered orally with 2000mg/kg of PPPE all dissolved in DMSO and Normal saline. Animals were weighed, anesthetized and blood samples were collected at the end 5 days of oral administration. Blood samples were collected by cardiac puncture for some biochemical parameters such as Total proteins, bilirubin, cholesterol, uric acid, creatinine, Urea and Uric acid activities as well as serum, AST and ALT were determined.ResultsAt the end of this research work, it was discovered that at doses higher than 500mg/kg there were no enzymatic leakages from the organs (kidney and liver) into the serum, compared to the control group.ConclusionThe study revealed that it is advisable to use doses tested on the animals for the management of insect pest on products meant to be consumed by human and for propagation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regular exercise reduces adipose tissue inflammation and improves glycemic control in Western diet‐fed mice despite hyperendothelinemia
Authors: Jurrissen Thomas J , Grunewald Zachary I , Ball James R , Ramirez‐Perez Francisco I , Woodford Makenzie L , Aroor Annayya R , Ayedun Lolade A , Winn Nathan C , Paradis Pierre , Schiffrin Ernesto L , Martinez‐Lemus Luis A , Padilla Jaume 
Published: June 2021

The upregulation of endothelin‐1 (ET‐1), a peptide primarily expressed and secreted into the circulation from endothelial cells, is a key molecular signature in obesity and type 2 diabetes. Given its potent pro‐oxidant and inflammatory effects, ET‐1 is thought to be implicated in the etiology of metabolic dysfunction. Based on previous evidence that regular exercise decreases ET‐1 expression and signaling, it is conceivable that the well‐established exercise‐induced anti‐inflammatory effects and associated improvement in glycemic control is, in part, mediated by a reduction in ET‐1 actions; however, this remains unknown. Accordingly, we tested the hypothesis that overproduction of ET‐1 from endothelial cells and resulting hyperendothelinemia limits the beneficial metabolic adaptations of exercise. Six‐week‐old male transgenic mice overexpressing human preproET‐1 selectively in the endothelium (eET‐1) and non‐transgenic (i.e., wild‐type) littermates were fed a Western diet (46% Kcal fat, 36% Kcal carbohydrate) to induce obesity, while they either remained sedentary or had free access to a running wheel for 12 weeks, creating four groups (n=7–12/group): wild‐type sedentary, wild‐type exercise, eET‐1 sedentary, eET‐1 exercise. Plasma ET‐1 was significantly elevated in eET‐1 mice (sedentary: 15.12±3.77 pg/ml; exercise 14.90±2.30 pg/ml) versus wild‐type (sedentary: 0.02±0.02 pg/ml; exercise: 0.29±0.14 pg/ml) in both sedentary and exercise‐trained mice. Exercise reduced final body weight, adiposity, fasting plasma insulin, HOMA‐IR, cholesterol and triglycerides compared to sedentary mice, and these effects were independent of genotype. Also independent of genotype, exercise reduced visceral adipocyte hypertrophy, immune cell infiltration, inflammatory gene expression (e.g., IL‐1β and CD11c), as well as enhanced visceral adipose tissue insulin signaling (i.e., increased ex vivo insulin‐stimulated pAkt/Akt). Furthermore, exercise improved glucose tolerance and this improvement was mildly enhanced in eET‐1 mice (i.e., significant genotype × exercise interaction). In summary, contrary to our hypothesis, we find that experimentally‐induced hyperendothelinemia does not impede exercise‐induced improvements in metabolic function, supporting the notion that reduced ET‐1 signaling is not required for the beneficial metabolic adaptations of exercise to manifest.Support or Funding InformationNational Institute of HealthThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PM2.5 Increases Lung Inflammation through the Sirtuin‐SREBF1‐inflammasome Axis
Authors: Tien Chia‐Ping 
Published: June 2021

Epidemiological studies have shown that particulate matter (PM2.5) is strongly associated with inflammatory lung disease, including bronchitis, asthma, chronic obstructive pulmonary disease (COPD), and lung cancer. However, the mechanisms remain unclear. To identify the molecular targets of PM2.5, we analyzed transcriptomics data by microarray chips in human pulmonary fibroblasts (HPF) and human bronchial epithelial cells (HBEpic) after exposure to PM2.5. Through Genespring software to normalization, we have selected candidate targets for prediction potential upstream regulator and pathway by Ingenuity Pathway Analysis (IPA) that may be mediated the induction of lung inflammation. Our results showed sterol regulatory element‐binding transcription factor 1 (SREBF1) was the most significant upstream transcription regular in both PM2.5 added HPF and HBEpic cells. Through RT‐PCR and western blotting analysis, we validated SREBF1 and its downstream inflammasome genes, including interleukin 6 (IL‐6), C‐X‐C motif chemokine ligand 8 (CXCL8), BCL2 related protein A1 (BCL2A1), and heme oxygenase 1 (HMOX1) were activated after exposure to PM2.5 in vitro. In addition, our results also indicated Sirtuin pathway was significantly suppressed after PM2.5 treatment. Sirtuin1 was found to directly deacetylate SREBF1 to inhibit its function in inflammasome activity, which can be blocked by the addition of PM2.5. Furthermore, complementary experiments were performed, including the addition of Sirtuin1 recombinant protein or shRNAs of SREBF1 to PM2.5 exposed normal lung epithelial cells and fibroblasts. The results revealed that the activation of inflammasome genes expression could be significantly reversed. In conclusions, our results identify the Sirtuin‐SREBF1‐inflammasome axis may play a significant role in PM2.5‐induced pulmonary diseases and may provide novel therapeutic targets for patients.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Macrophage digesting dysfunction by Angiotensin II through lysosomal cathepsin B‐mediated Nlrp3 inflammasome activation
Authors: Lian Dawei , Lai Jieqing , Wu Yanjiao , Wang Lei , Yao Lin , Zhang Yang , Huang Yi , Chen Yang 
Published: June 2021

Angiotensin II (Ang II) is an octapeptide hormone that plays a significant role in mediating hypertension. Hypertension is considered a chronic inflammatory disease; however, the molecular basis underlying the sterile inflammatory response involved in the process of hypertension remains unclear. The present study aims to investigate the role of macrophage Nlrp3 inflammasomes in engulfing and digesting microbes, which are associated with macrophage function, an early onset of hypertension‐associated macrophage injury. Our findings demonstrate that AngII stimulation decreased the NO release and macrophage digestion in cultured RAW264.7 cell. Accompanied with these changes, AngII markedly increased the Nlrp3 inflammasome formation and activation. As AngII‐induced up‐regulation of Nlrp3 inflammasome in macrophage was attributed to lysosome dysfunction induced cathepsin B release. In addition, Losartan, a nonpeptide Ang II receptor antagonist, decreased AngII‐induced Nlrp3 inflammasome activation, lysosomal membrane permeability, lysosomal cathepsin B release, and macrophage digestion dysfunction in macrophage. Similarly, AngII induced macrophage digestion microbes and NO production, which were blocked by Nlrp3 gene silencing and ATI gene silencing. It is concluded that Angiotensin II enhanced lysosomal membrane permeabilization and consequent release of lysosomal cathepsin B, resulting in activation of the macrophage Nlrp3 inflammasome, which may contribute to NO mediated the development of digesting microbes dysfunction.Support or Funding InformationThis study was supported by the National Natural Science Foundation of China (81603587 and 81603668) and by the Guangdong Science and Technology Department (2017A020211016).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction between the Dopamine D4 Receptor and the Angiotensin II Type 1 Receptor in Rat Astrocytes
Authors: O'Connor Ann Tenneil , Clark Michelle A. , Trivedi Malav 
Published: June 2021

ObjectiveTo determine the basal expression of the Dopamine D4 receptor (D4R) in Wistar rat astrocytes and investigate if Angiotensin (Ang) II can regulate D4R expression.BackgroundThe interaction between the renin angiotensin system (RAS) and the dopaminergic system in the regulation of peripheral blood pressure is well established; however, such interaction in the brain is not studied. Our goal is to investigate whether there is an interaction between these two systems in a rat model brain system.MethodsBrainstem and cerebellum astrocytes were prepared from 2–3 days Wistar rat pups. These brain regions were selected because they express RAS components. Quiescent cells were treated with 100nM Ang II for 1 to 48 hours. We determined D4R and AT1R, protein levels by western blots and mRNA levels by quantitative real time polymerase chain reaction.ResultsWistar cerebellum astrocytes had significantly higher basal D4R protein levels compared to Wistar brainstem. Basal D4R mRNA levels in cerebellar astrocytes were higher than the basal D4R mRNA levels in brainstem astrocytes. Basal AT1R protein expression was significantly higher in brainstem astrocytes compared to cerebellar astrocytes. Ang II reduced the D4R protein levels relative to untreated controls in Wistar brainstem at most of the time points examined. There were no significant differences in D4R protein levels in cerebellum astrocytes, relative to controls after Ang II treatment.ConclusionWe demonstrated the ability of Ang II to reduce the D4R protein levels in Wistar rat brainstem samples. Our results showed that under physiological conditions, D4Rs in brainstem astrocytes were more sensitive to Ang II treatments compared to those in cerebellum. This could be attributed to the role of the brainstem in blood pressure regulation. Indeed, brainstem astrocytes had higher AT1R protein levels compared to cerebellum astrocytes. This interaction between the AT1R and the D4R is a potential therapeutic target for pathophysiological conditions mediated by these two systems. Further studies will investigate this interaction in astrocytes and neurons isolated from spontaneously hypertensive rats, a rat model of Hypertension and Attention Deficit Hyperactivity Disorder.Support or Funding InformationThis study was funded through a President's Faculty Research and Development Grant from Nova Southeastern 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.
Anti‐inflammatory effect of wood‐cultivated ginseng in LPS‐stimulated RAW264.7 cells
Authors: JEONG JIN BOO 
Published: June 2021

Ginseng (Panax ginseng) has been reported to exert an anti‐inflammatory activity in a variety of inflammatory. However, inflammation‐regulatory activity of wood‐cultivated ginseng has not been thoroughly evaluated. In this study, we evaluated the anti‐inflammatory effect of wood‐cultivated ginseng and elucidated the potential mechanisms in LPS‐stimulated RAW264.7 cells. Inhibitory effects of the old wood‐cultivated ginseng (WCG‐O), young wood‐cultivated ginseng (WCG‐Y) and ginseng (G) on NO and PGE2 production were examined using the Griess assay and ELISA kit. Suppressive effects of WCG‐O on inflammatory gene expression, transcriptional activation, and inflammation signaling events were investigated using Western blot analysis, RT‐PCR analysis and luciferase activity reporter gene assay. WCG‐O dose‐dependently suppressed nitric oxide (NO) and Prostaglandin E2 (PGE2) production in LPS‐stimulated RAW264.7 cells. In addition, WCG‐O attenuated LPS‐mediated overexpression of iNOS and COX‐2. In addition, WCG‐O blocked the expression of TNF‐α and IL‐1β in LPS‐stimulated RAW264.7 cells. In elucidation of the potential mechanisms for anti‐inflammatory effect, WCG‐O inhibited the activation of IκK‐α/β, the phosphorylation of IκB‐α, and degradation of IκB‐α, which results in the inhibition of p65 nuclear accumulation and NF‐κB activation. In addition, WCG‐O suppressed the activation of ERK1/2, p38 and JNK, which results in the inhibition of ATF2 nuclear accumulation. These results indicate that WCG‐O may exert anti‐inflammatory activity through the inhibiting NF‐κB and MAPK signaling. From these findings, WCG‐O has potential to be a candidate for the development of chemoprevention or therapeutic agents for the inflammatory diseases.Support or Funding InformationThis research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF‐2016R1D1A3B03931713).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.