bluuebunny/crossref_metadata_2025 huggingface dataset:
Glycemic and Insulinemic Response to Ingestion of a Novel Food Bar Containing Whey Protein and Isomalto‐Oligosaccharides
Authors: Grubic Tyler Joseph , Kreider Richard B , Sowinski Ryan , Dalton Ryan , Favot Christopher , Greenwood Mike , Earnest Conrad P , Reyes Aimee , Rasmussen Chris
Published: June 2021

This study examined the glucose and insulin response of ingesting whey protein bars using a plant fiber (isomalto‐oligosaccharides, IMO) as the carbohydrate source. In a randomized and crossover manner, 10 healthy men and women (4 female, 6 male; BMI <24.9 kg/m2) donated fasting blood samples prior to ingesting two food bars (FB) each containing 20 g of a whey protein blend, 25 g of carbohydrate (13 g IMO, 4 g sugar, 8 g fiber), and 7 g of fat (1.5g saturated) or 50 g of dextrose (PLA). The experiment was repeated 7 to 10 days later while ingesting the alternative treatment. Blood samples were taken at 10, 20, 30, 60, 90, and 120 min post‐ingestion while subjective ratings related to appetite and hypoglycemia were obtained at 0, 60 and 120 min. Data were analyzed by general linear model statistics and are presented as mean [95% CI] changes from baseline. Results revealed that the glycemic response to ingestion of the FB was significantly lower during the first 60 min following ingestion in comparison to the dextrose PLA. Glucose and insulin levels peaked 30 minutes after ingestion; interestingly FB ingestion only marginally increased glucose from baseline for ten minutes. The overall AUC for glucose was significantly lower in FB treatment (530 ± 48, PLA 697 ± 67 mmol‐h/L, p<0.001). No significant differences were observed between treatments in the overall insulin AUC (FB 4,185 ± 1,934, PLA 3,888 ± 707 μIU/mL‐h/L, p=0.65). The glucose integrated AUC (iAUC) change from baseline was significantly lower with FB ingestion (FB 65 [49, 82], PLA 209 [170, 244] mmol‐h/L, p<0.001) while no differences were observed between treatments in insulin iAUC responses. In comparison to the dextrose standard, the FB had an iAUC derived glycemic index (GI) of 34 [CI 23, 46] and a glycemic load (GL) of 8.5 [CI 5.6, 11.6]. Participants also reported significantly less subjective ratings of appetite and hunger and greater satisfaction from food and feeling of fullness in both studies. No significant differences over time or between treatments were observed in ratings of symptoms of hypoglycemia. Results indicate that ingestion of a whey protein bar using IMO as the source of carbohydrate elicited a low glycemic response in comparison to a reference carbohydrate in healthy individuals. Thus, this FB may serve as a low glycemic food option for individuals on a low glycemic diet and/or athletes interested in optimizing nutrient availability around exercise.

Support or Funding Information

Exercise and Sport Nutrition Lab, Human Clinical Research Facility, Department of Health & Kinesiology, Texas A&M University, College Station, Texas, USA

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structural Characterization of Vitiligo Associated T Cell Receptor: Towards the Development of Improved Melanoma Immunotherapy
Authors: Smith Angela R , Eby Jonathan M , Ankney Christopher M , Cosgrove Cormac , Henning Steven W , Nishimura Michael I , Le Poole Caroline , Baker Brian M
Published: June 2021

Melanoma‐associated vitiligo is a notable example of the intersection between tumor immunity and autoimmunity. Normal melanocytes and melanoma cells present many of the same differentiation antigens which may explain why vitiligo can develop as a side effect in melanoma patients with active tumor immune responses. Most interestingly, it has been shown that vitiligo‐derived T cells can kill melanoma cells, suggesting that vitiligo may be a useful source of T cells for adoptive T cell therapy against melanoma.

T cell receptors (TCRs) are responsible for the antigen specificity of T cells and are the major component of new cell‐based immunotherapies. TCRs are restricted to the recognition of antigen peptides presented by major histocompatibility complex (MHC) proteins. The SILv44 TCR was isolated from a vitiligo patient's skin infiltrating T cells and has shown superior inhibition of tumor growth in a mouse model. A structural and biophysical characterization of SILv44 compared to two melanoma‐derived TCRs (R6C12 and T4H2) that recognize the same peptide‐MHC (gp100209‐2M/HLA‐A2) will allow us to assess the TCR properties best associated with anti‐melanoma immunity. Interestingly, all three TCR transduced CD8 T cells elicited different cytokine expression profiles. Despite possessing the strongest anti‐tumor immunity, SILv44 has the weakest affinity for its antigen compared to R6C12 and T4H2. When expressed in primary CD8 T cells and exposed to antigen‐presenting cells, all receptors elicited interferon‐gamma (IFN‐γ) responses which correlated with TCR affinity and inversely correlated with tumor growth suppression. These results suggest that TCR affinity and IFN‐γ secretion may not be the key factors in productive T cell recognition of shared tumor antigens. The SILv44 crystal structure indicates that the global positioning of SILv44 over this peptide‐MHC (pMHC) is similar to other HLA‐A2 bound TCR complexes. SILv44 predominantly uses its β chain to engage this pMHC. Crystal structure determinations of R6C12 and T4H2 complexes are in progress and will be compared to the SILv44 complex.

Support or Funding Information

Walther Cancer Foundation Interdisciplinary Interface Training Project

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of HDAC8 Ameliorates Pulmonary Fibrosis
Authors: Saito Shigeki , Zhuang Yan , Lasky Joseph
Published: June 2021

Idiopathic pulmonary fibrosis (IPF) is a fibroproliferative lung disease with high mortality, and fibroblast‐myofibroblast differentiation (FMD) is thought to be a key event in the pathogenesis of IPF. Histone deacetylase 8 (HDAC8) has been shown to associate with α‐smooth muscle actin (α‐SMA; a marker of FMD) and regulates cell contractility in vascular smooth muscle cells. However, the role of HDAC8 in FMD or pulmonary fibrosis has never been investigated. This study investigated the role of HDAC8 in pulmonary fibrosis, with a focus on FMD. We observed that HDAC8 expression was increased in IPF lung tissue as well as TGF‐β1‐treated normal human lung fibroblasts (NHLFs). Our immunoprecipitation experiments revealed that HDAC8 associated with α‐SMA in TGF‐β1‐treated NHLFs. HDAC8 inhibition with NCC170 (HDAC8‐ selective inhibitor) repressed TGF‐β1‐induced fibroblast contraction and α‐SMA protein expression in NHLFs cultured in collagen gels. HDAC8 inhibition with HDAC8 siRNA also repressed TGF‐β1‐induced expression of CTGF and PAI‐1, and increased PPAR‐γ expression in NHLFs. Chromatin immunoprecipitation quantitative PCR (ChIP‐qPCR) using an antibody against H3K27ac (histone H3 acetylated at lysine 27; a known HDAC8 substrate and a marker for active enhancers) suggested that HDAC8 inhibition with NCC170 ameliorated TGF‐β1‐ induced loss of H3K27ac at the PPAR‐γ gene enhancer. Furthermore, NCC170 treatment significantly decreased fibrosis measured by Ashcroft score as well as type‐1 collagen expression in bleomycin‐treated mouse lungs. In conclusion, these data suggest that HDAC8 contributes to pulmonary fibrosis and that there is a therapeutic potential for HDAC8 inhibitors to treat IPF as well as other fibrotic lung diseases.

Support or Funding Information

Wetmore Foundation: Shigeki Saito M1: 555007G1; Louisiana Clinical and Translational Science (LA CaTS) Center: Shigeki Saito 1 U54 GM104940

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Long‐term endurance exercise coactivates autophagy/mitophagy and anabolic signaling
Authors: Quintin Stephan , Jang Yongchul , Kwon Insu , Song Wankeun , Cosio‐Lima Ludmila M. , Lee Youngil
Published: June 2021

Growing evidence has identified that autophagy plays an important role in sustaining normal cellular function. Endurance exercise (EE) is a potent inducer of autophagy in multiple tissues. However, the exact underlying signaling pathways responsible for exercise‐induced autophagy in brain has not been fully understood yet. In this study, we investigated potential signaling pathways involved in EE‐induced autophagy and mitochondrial dynamics in the cortex of the brain. C57BL/6 male mice were randomly assigned to a control group (CON, n=12) and an endurance exercise group (EXE, n=12). Our data demonstrated that EE‐induced autophagy evidenced by upregulation of autophagy‐related proteins (e.g., LC3 II, BECLIN1, ATG7, LAMP2, CATHEPSIN L and TFEB) coincides with increased anabolic signaling state (p‐AKT, p‐mTOR and p‐p70S6K). Furthermore, we revealed that EE promoted mitophagy evidenced by increases in LC3 II, p62, BNIP3, PINK, DRP1 in isolated mitochondrial fraction). We attempted to examine if low degree of oxidative stress would be a potential factor involved in EE‐induced autophagy. As opposed to our postulation, oxidative stress was not the case since EE did not increase protein oxidation, but rather enhanced antioxidative capacity (CATALASE and PRX III). Taken together, our results indicate that EE‐induced autophagy/mitophagy concomitant with anabolic signaling activation may be a novel protective signaling that mediates facilitation of healthy brain.

Support or Funding Information

This project was supported by a research grant from the University of West Florida though Office of Research and Sponsored Programs (R0062) and UWF Florida Research Fellowship to YL (CF6672 and CR0070).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Electromyostimulation with Blood Flow Restriction Enhances Activation of mTOR and MAPK Signalling Pathways in Rat Gastrocnemius Muscles
Authors: Natsume Toshiharu , Yoshihara Toshinori , Naito Hisashi
Published: June 2021

Aim

Neuromuscular electrical stimulation (NMES) combined with blood flow restriction (BFR) can induce muscle hypertrophy. However, cellular mechanisms underlying the muscle hypertrophy induced by NMES combined with BFR remain unclear. Thus, the purpose of this study was to elucidate the signalling pathways involved in NMES with BFR induced muscle hypertrophy.

Methods

Age‐matched male Wistar rats (6 months old, n=7 per group) were assigned randomly into control, BFR alone (BFR), NMES alone (NMES) and NMES combined with BFR (NMES/BFR) groups. NMES induced 25 isometric contractions lasting 8 s with a 4 s resting period between each contraction in the gastrocnemius muscle. Four sets in total were performed, with 1‐min intervals between each set. Latex cuff was placed on the proximal portion of hind limb and BFR was conducted in four sets (each set 5 min) with 1‐min rest intervals in between at 200 mmHg. Venous blood was collected from the lateral tail vein to determine pH, H+, lactic acid concentration before and immediately after the treatments. Expression levels of proteins related to muscle hypertrophy were determined by western blot analysis.

Results

The application of NMES combined with BFR encouraged muscle fatigue more than NMES alone. NMES/BFR induced greater changes in accumulation of metabolites and increase in the weight of gastrocnemius muscle. The phosphorylation of mTOR and MAPK signalling related proteins also enhanced following NMES combined with BFR, compared with other conditions.

Conclusions

We elucidated that NMES enhanced the activation of mTOR and MAPK signalling pathways when combined with BFR.

Support or Funding Information

This work was supported by Japan Society for Promotion of Science (JSPS) KAKENHI Grant No. 16K16564 (T. Natsume) and Grant No. 17K01765 (T. Yoshihara). Grants from the MEXT‐Supported Program for Strategic Research Foundation at Private Universities also supported this research.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inter‐hemispheric analysis of compensatory neural activity in deep brain stimulation patients
Authors: Uy Daniel R , Thompson John R , Abosch Aviva
Published: June 2021

Parkinson's disease is a progressive neurodegenerative disorder that is chiefly marked by the progressive loss of dopaminergic neurons, resulting in motor and some non‐motor (e.g., olfactory dysfunction) symptoms (Calabresi et al., 2015). The motor related symptoms are a result of the reduction of dopamine caused by the decline in the population of dopamine producing neurons in the substantia nigra of the basal ganglia. Deep brain stimulation (DBS) is a symptom management treatment for pharmaceutically unmanageable Parkinson's disease because of its ability to improve motor functions and thus quality of life (Martinez‐Ramirez et al., 2015). Individuals will typically have two procedures, one hemisphere per surgery, and thus have electrodes implanted in both hemispheres of the brain. Given the necessity for electrophysiological evaluation of implant target, the DBS procedure provides one of the few opportunities to examine changes in neural networks in the human brain. This study sought to examine whether there were compensatory changes in neural activity within basal ganglia structures as a function of surgical disruption. The current results suggest that there is a compensatory increase in contralateral neural activity within basal ganglia structures following surgery.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
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 Information

National 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 Information

NIH, The Jayne Koskinas Ted Giovanis Foundation for Health and Policy

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating 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

Objectives

The 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.

Methodology

Piper 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.

Results

At 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.

Conclusion

The 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 Information

National Institute of Health

This 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 Information

This 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

Objective

To determine the basal expression of the Dopamine D4 receptor (D4R) in Wistar rat astrocytes and investigate if Angiotensin (Ang) II can regulate D4R expression.

Background

The 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.

Methods

Brainstem 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.

Results

Wistar 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.

Conclusion

We 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 Information

This 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 Information

This 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.
Lidocaine blocks the proliferation, Migration, and Epithelial‐Mesenchymal Transition of Human Retinal Epithelial cells
Authors: Kim Yeon A , Kwon Yoon Hyung , Woo Min Seok
Published: June 2021

Introduction

Proliferative vitroretinopathy (PVR) is a major cause of failure in surgery of rhegmatogenous retinal detachment (RRD). The incidence of PVR approximately attains up to 5~10% of RDD and 75% of redetachment after surgery, however, the strategy for precautions or resolving the PVR is very limited. Lidocaine is well known and widely used local anesthetics. We evaluated the effects of lidocaine on epithelial‐mesenchymal transition (EMT) and migration in retinal pigment epithelial cells, which is major cell type involved in the development of PVR, and its mechanism.

Methods

Adult retinal pigmented epithelial 19 (ARPE 19) cell line was used and MTT (3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide) assay was performed with various concentrations of lidocaine. We induced ARPE to EMT by TGFβ‐1 and it was demonstrated with western blot of α‐SMA (smooth muscle actin), vimentin, E‐cadherin, and collagen I. The inhibitory effects of lidocaine in TGFβ‐1 signaling were also evaluated with western blot of ERK (extracellular signal regulated kinase), PI3K (Phosphoinositide 3‐kinase), Akt (Protein kinase B), and smad. The change of motile activity by TGFβ‐1 and lidocaine was demonstrated with migration assay.

Results

Lidocaine was inhibited the proliferation of ARPE cells in concentration dependent manner after treatment of lidocaine, 24 hours and 48 hours, respectively. Migration activity of ARPE cells were inhibited by lidocaine and the migration of ARPE cells undergoing EMT by TGFβ‐1 was also inhibited significantly. Lidocaine prevented EMT by decreasing the activations of ERK, PI3K, Akt and smad, result in decreasing the expression of α‐SMA, vimentin, and collagen I synthesis and increasing the expression of E‐cadherin.

Conclusions

Lidocaine blocks the proliferation, migration, and EMT of ARPE cells by inhibition of TGFβ‐1 signaling pathway.

Support or Funding Information

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean Government (NRF‐2017R1C1B5017925).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of Curcumin on P‐glycoprotein Function
Authors: PENG CHI‐WEN , Teng Yu‐Ning , Hung Chin‐Chuan
Published: June 2021

Multidrug resistance (MDR) is one of the leading cause of cancer treatment failure. P‐glycoprotein (P‐gp) has been reported as a major mechanism of MDR that increased the efflux of drugs. Three generations of P‐gp inhibitors have been developed, however, systemic toxicities limited the clinical application of these compounds. Therefore, identification of the potent and non‐toxic P‐gp inhibitors from natural products may be an alternative choice. The objective of this study was to investigate the inhibitory effect and molecular mechanism of curcumin on human P‐gp efflux function.

HEK293 cell were stably transfected with human P‐gp. The MDR1 shift assay was applied to evaluate whether curcumin is a substrate of P‐gp. The effect of curcumin on P‐gp function was evaluated by rhodamine 123 accumulation assay and calcein AM uptake assay. Rhodamine 123 efflux assay was performed to analyze the inhibitory mechanism.

Results from cytotoxicity assay and MDR1 shift assay indicated that curcumin is a safe natural product without cytotoxicity and not a P‐gp substrate. The intracellular fluorescence of rhodamine 123 was higher in curcumin treated P‐gp expressed cells than no‐treatment control, suggesting curcumin may inhibit P‐gp efflux function. Results of calcein‐AM uptake assay and rhodamine 123 efflux assay demonstrated that curcumin significantly inhibited P‐gp efflux function in dose dependent manner. Furthermore, curcumin uncompetitively inhibited rhodamine 123 efflux by P‐gp.

This study demonstrated that curcumin, sourced from natural plants, as a potential P‐gp function modulator. Although the exploring of novel inhibitors from natural products is still in the early stages, the promising beginning and detailed mechanism investigation provided by this study will be helpful in the future in vivo studies and clinical use.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AKBA exerts anti‐tumor effects in glioblastoma cells by arresting cell cycle at G2/M phase
Authors: WANG JINHUA , Li Wan , Lyu Yang , Du Guanhua
Published: June 2021

Glioblastoma (GBM), accounting for about 50 % of all gliomas, is the most common, malignant, and lethal primary brain tumor in adults. Up to now, there is no effective drug for GBM. 3‐O‐Acetyl‐11‐keto‐β‐boswellic acid (AKBA) is one of main active ingredients of the gum resin of Boswellia serrata and Boswellia carterri Birdw. AKBA was reported to inhibit the growth of many cancer cells; however, the precise molecular mechanisms of its anti‐cancer activities in GBM remain unclear. Here, we found that treatment of AKBA in U251 and U87‐MG glioma cells inhibited the cell proliferation, released the cellular LDH, decreased the DNA synthesis, and inhibited the migration, invasion, and colony formation of cells. AKBA also increased the apoptotic rate of cells, decreased the mitochondrial membrane potential, inhibited the activity of caspase 3/7, and decrased the protein expreesion of cleaved‐caspase 3, cleaved PARP in U251 and U87‐MG cells. RNA‐Sequencing analyses showed that AKBA suppressed the expression of pRB, FOXM1, Aurora A, PLK1, CCD25C, p‐CDK1, cyclin‐B1, Aurora B, and TOP2A while increased the expression of p21 and GADD45A, which was further validated by qRT‐PCR and Western blot analyses. These results demonstrated that AKBA induced mitochondrial dependent apoptosis in glioblastoma cells which was associated with arresting the cell cycle at G2/M phase by regulating the p21/ FOXM1/cyclin B1 pathway and inhibiting the mitosis of glioma cells by downregulating the Aurora B/ TOP2A pathway. Taken together, our findings suggest that AKBA might be a promising chemotherapy drug in the treatment of GBM.

Support or Funding Information

National Natural Science Foundation of China (N0. 81573454 for JINHUA WANG, N0. 81703536 for WAN LI and 81703565 for WEIQI FU), CAMS Innovation Fund for Medical Sciences (CIFMS)(2016‐I2M‐3‐007) and Natural Science Foundation of Beijing (7172142).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Isolation, Characterization, and Synthesis of Cuticular Hydrocarbon Natural Products from Drosophila athabasca, A Species of Evolutionary Interest
Authors: Shuler John F , Lin Nar , Gay Brittany L. , Yukilevich Roman , Kehlbeck Joanne D
Published: June 2021

Drosophila athabasca, a species complex native to North America, provides a unique opportunity to study nascent speciation arising from sexual isolation. Some have suggested that cuticular hydrocarbons (CHCs) influence the sexual selection process. Extensive analyses have resulted in complex understandings of CHC involvement in insect mate preference, with minimal work focusing on D. athabasca. The goal of this study is to identify, synthesize and measure biologically relevant quantities of CHCs present in this species complex. Compilations of synthetic material will allow for biological evaluation and a greater understanding of CHC function in D. athabasca. Gas chromatography ‐ mass spectrometry (GC‐MS) from several isofemale lines of D. athabasca reveal a wide array of functionalities, including: saturated, unsaturated, and branched alkanes, fatty acids, acetates, and esters. Dimethyl disulfide (DMDS) derivatization led to unambiguous determination of positions of unsaturation. CHC identities were confirmed through comparison to synthetic standards. To date, over 100 CHCs have been characterized in D. athabasca using previously described analytical and organic methods. An overview of the analyses and syntheses, along with future directions, will be presented.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synthesized‐Chlorotoxin‐Conjugated Cytochrome C as a Potential Drug for Targeting Glioma
Authors: Delinois Louis Jean , Ramirez‐Paz Josell , González‐Feliciano Jose A. , Maldonado‐Hernández Rafael , Barcelo‐Bovea Vanessa , Joaquin Ovalle Freisa M. , Fulvio Pasquale F. , Griebenow Kai
Published: June 2021

Amongst primary brain tumors, gliomas are considered the most lethal malignant tumors due to their resistance to radiation and chemotherapy and the difficulty to accurately localize them within the tissue. Chlorotoxin (Ctx), a small neurotoxin of 36 amino acids isolated from the venom of the Israeli scorpion Leiurus quinquestriatus, binds specifically and with high affinity a 72 kDa chloride channel membrane receptor in human gliomas and was proposed to cross the Blood Brain Barrier. These channels are reported to be absent or in low abundance in healthy tissues or in tumors of non‐glial origin. As a component of the scorpion venom, Ctx induces paralysis in small insects or other invertebrates however, no apparent signs of toxicity have been observed when injected in vertebrates. Cytochrome c (CytC), a small membrane impermeable mitochondrial protein (MW= 12 KDa) that facilitates electron transport in cellular respiration, is also known to mediate apoptosis, a normal process of controlled cell death, through the activation of a caspase cascade. In this work, a chemically synthesized‐Ctx is covalently conjugated to CytC through reversible disulfide bridges to accomplish targeted delivery to a glioma cell line 9L/lacZ from Rattus norvegicus and NIH 3T3 from Mus Musculus. Size exclusion chromatography and mass spectrometry characterization of the Ctx‐CytC conjugate show that the Ctx was successfully attached to CytC. Preliminary results from cell viability essay reveal that the Ctx‐Cytc was 40% more toxic than Ctx alone and more toxic to 9L/lacZ than the normal cell line NIH/3T3. These results suggest that the Ctx‐CytC conjugate can be further developed in order to be considered as an alternative potential targeted drug for treatment of gliomas.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.