bluuebunny/crossref_metadata_2025 huggingface dataset:
EXPRESSION OF IRISIN AND ITS RELATION WITH ANTHROPOMETRIC AND BIOCHEMICAL PARAMETERS IN DIABETES MELLITUS TYPE II PATIENTS
Authors: Mendonça Maria Isabel Sacchi , Mathia Vanessa Lopes , Simões Diogo Pimenta , Perez Matheus Moreira , Bacci Marcelo Rodrigues , Murad Neif , Encinas Jessica Araújo , da Costa Aguiar Alves Beatriz , Fonseca Fernando Luiz Affonso , da Veiga Glaucia Luciano
Published: June 2021

Aim

To evaluate the gene expression of irisin in patients with type 2 diabetes mellitus (DM2) and its possible involvement in the metabolic changes and obesity in this pathophysiological situation.

Methods

Patients anthropometric data were obtained, as well as samples of blood were collected in the clinic of specialties from Faculty of Medicine of ABC. A list of healthy individuals CT (n=14) and DM2 patients (n=16) diagnosed for less than 10 years was obtained. The criteria for the diagnosis of DM2 were the one used by the American Diabetes Association (ADA). Volunteers filled in terms of consent, and all of them were greater than or equal to 21 years old. After collection, it was evaluated complete hemogram and biochemical parameters: glucose and glycated hemoglobin (A1C). The expression of molecular biomarker irisin in peripheral blood leukocytes was performed by RT‐qPCR technique. Results were expressed as mean ± standard deviation (SD) and analyzed by the program GraphPad Prism® 6.0, using the Student's t‐test and Mann‐Whitney test. As for the analyzes of correlation, it was done the Spearman correlation test. The significance level was 5% (descriptive value *p<0.05).

Results

The mean age of the patients studied was 51±15 years for the CT group (n=14) and 64±10 years for the DM2 (n=16). As for the biochemical parameters, the following results were obtained: glucose (CT 92±13, n=14 DM2 versus 143±40 mg/dL, n=16 *p<0.05); A1C (CT 5.6±0.5, n=14 DM2 versus 7.1±1.6%, n=16 *p<0.05); irisin gene expression in peripheral blood was obtained by the formula 2^‐ΔCT and values found in CT group were 8.506e‐006±1.412e‐005, n=6. The ones found in DM2 group were 0.02288± 0.08050 n=14, p=0.06. The correlation test between the expression of irisin versus Body Mass Index (BMI) in DM2 was r=0.5221, p=0.06. There was no correlation between the glucose levels and A1C with irisin expression.

Conclusion

Despite the preliminary findings, the data showed a tendency toward a positive correlation between the expression of irisin and BMI in diabetic patients. These data suggest that irisin is associated to possible changes in adiposity which happen in obesity associated to DM2.

Support or Funding Information

Financial Support: CAPES, CNPq.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Quantifying the Contribution of Cardiomyocyte Metabolic Dysfunction to the Heart Mechanical Function
Authors: Lopez Rachel , Gao Xin , Van den Bergh Françoise , Beard Daniel
Published: June 2021

Hydrolysis of ATP provides the chemical free energy to drive the molecular processes underlying cardiac pumping. Under normal conditions the heart hydrolyzes and resynthesizes the entire ATP pool several times per minute. In the failing heart, cardiomyocyte metabolic dysfunction leads to a reduction in the chemical potential (free energy of ATP hydrolysis) at which ATP is synthesized and is made available to drive cellular processes. We hypothesize that this metabolic/energetic dysfunction causes contractile dysfunction of the myocardium in heart failure, contributing to the whole‐body phenotype (venous congestion, exercise intolerance) of heart failure in humans. To test this hypothesis, we used a rat transverse aortic constriction (TAC) model of cardiac decompensation/heart failure to measure the relationships between functional parameters (cardiac output, ejection fraction, etc.) and myocardial energetic state, determined based on measurements of myocardial metabolite pools and mitochondrial oxidative capacity. Cardiac function was measured by echocardiography and hearts were harvested for mitochondrial isolation and phosphate metabolite measurements at the end‐point, either 15 weeks post‐surgery, or 18 weeks post‐surgery. Cardiac phosphate metabolites levels were measured using NADH dependent enzymatic assays and mitochondrial oxidative capacity was measured using an Oroboros O2k Oxygraph. In both cohorts of rats, there was an overall reduction in fatty acid and carbohydrate oxidative capacity, with an increase in the ratio of maximal carbohydrate to fatty acid oxidative capacity in the 18‐week TAC rats compared to sham controls. In addition, we also observed differences in phosphate metabolite levels in both 15‐week TAC and 18‐week TAC rats compared to controls and correlations between metabolite mechanical functional changes. Furthermore, we used multi‐scale computational models of myocardial metabolism and metabolic state‐dependent contractile dynamics to quantify the contribution of reductions in oxidative capacity and metabolite pools levels to heart mechanical function. Our results suggest that there is a causal link between a reduction in free energy of ATP hydrolysis and kinetic impairment of the myosin ATPase cross‐bridge cycle in decompensated hypertrophy/heart failure.

Support or Funding Information

Funded by NIH Grants 5T32GM008322‐28 and U01HL122199

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Control of Cardiac Mitochondrial Fuel Selection by Calcium
Authors: Jones Edith , Van den Bergh Françoise , Kandel Sunil M. , Dasika Santosh K. , Dash Ranjan K. , Beard Daniel A.
Published: June 2021

Calcium ion concentration modulates the function of several mitochondrial enzymes. Specifically, the kinetic operations of the decarboxylating dehydrogenases pyruvate dehydrogenase, isocitrate dehydrogenase, alpha‐ketoglutarate dehydrogenase are all affected by [Ca2+]. Previous studies have shown that, despite its ability to affect the function of specific dehydrogenases, [Ca2+] does not substantially alter mitochondrial ATP synthesis in vitro or in vivo in the heart. We hypothesize that, rather than contributing to respiratory control, [Ca2+] plays a role in contributing to fuel selection. Specifically, cardiac mitochondria are able to use different primary carbon substrates (carbohydrates, fatty acids, and ketones) to synthesize ATP aerobically in the living cells. To determine if and how [Ca2+] affects the relative use of carbohydrates versus fatty acids in vitro we measured oxygen consumption and TCA cycle intermediate concentrations in suspensions of cardiac mitochondria with different combinations of pyruvate and palmitoyl‐L‐carnitine in the media at various [Ca2+] and ADP infusion rates. Stoichiometric analysis of the data reveals that when both fatty acid and carbohydrate substrates are available, fuel selection is sensitive to both the rate of ADP infusion and to the calcium concentration. With increasing both the rate of oxidative ATP synthesis and [Ca2+], the fuel utilization ratio shifts to increased fractional consumption of pyruvate. Thus, our results indicate that the effects of ATP synthesis rate and [Ca2+] are interdependent.

Support or Funding Information

This work supported by NIH grants HL122199 and T32GM008322

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Adaptive effects of prolonged mitochondrial‐targeted catalase on adipocyte differentiation
Authors: Yang Sophia , Esterly Harrison , Tavares Kendall , Christian Brooke
Published: June 2021

One cause of obesity is an increase in the total number adipocytes by hyperplasia. To increase fat cell number, adipocyte precursors undergo differentiation into mature fat cells, a process which requires reactive oxygen species (ROS). ROS are generated in the mitochondria during oxidative phosphorylation, and previous studies have shown that short‐term treatment with mitochondria‐targeted antioxidants can reduce adipogenesis. This suggests that antioxidant therapy might be useful to treat obesity, although the efficacy of antioxidants for long‐term treatment has not been investigated. Mice that have catalase targeted to mitochondria (mCAT mice) have increased levels of the antioxidant enzyme throughout life. We tested whether mCAT mice would display reduced fat pad mass or body weight due to a reduction in mitochondrial hydrogen peroxide. At 12 weeks of age, mCAT mice showed no significant decrease in size or weight of subcutaneous fat depots compared to wild‐type mice, which suggests that mCAT pre‐adipocytes were able to bypass the requirement for ROS and differentiate normally. To test this, we isolated preadipocytes from subcutaneous fat pads of mCAT mice and compared their differentiation profiles to cells transiently transfected with a plasmid encoding mitochondrial catalase. As expected, short‐term antioxidant treatment resulted in little to no differentiation. Although preadipocytes from mCAT mice differentiated more slowly in culture, they were ultimately able to reach full differentiation. Gene expression profiles of regulatory proteins that function during differentiation in cells with short‐term antioxidant treatment were different than those with long‐term exposure. Current work involves understanding how these gene expression changes are directly affected by mitochondrial hydrogen peroxide.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
That's Swell! The Role of Aquaporin in an Inflammatory Response
Authors: Arnholt Mark , Betts Lilly , Hawkins Abigail , Herro Madeleine , Hoefs Alisha , Powers Farren , Sill Brooke , Munzenmaier Diane H
Published: June 2021

Death due to lung inflammation has increased 31 percent over the last 35 years. A protein that influences this mortality rate is aquaporin‐4. This tetramer protein is a water channel that is water permeable. This integral protein, made up of six transmembrane helices and two pore helices, creates an hourglass shape. The structure of aquaporin is vital to its function due to pore helices preventing large molecules from passing through the membrane, only facilitating the movement of water molecules or small solutes. These two pore helices allow one molecule to pass at a time, making the net water movement facilitation easier. All types of aquaporins have NPA (asparagine‐proline‐alanine) motifs which are located in the central channel and are responsible for water permeation and intracellular processing. In aquaporin‐4, there are two NPA motifs at residues 97–99 and 213–215. Scientists discovered that aquaporin's response to bacterial stimuli can alter its pore size. Trauma, toxins or injury to tissue can cause a cell to become hypotonic. Aquaporin‐4 is specifically related to lung injury and inflammation because it is highly expressed in humans. The HUHS MAPS Team (Modeling A Protein Story) has designed a model of the wild form of AQP4 with 3D printing technology to investigate structure‐function relationships. The extensive research on aquaporins in relation to lung inflammation can contribute to the scientific and medical world and eventually target new pharmacological therapies.

Support or Funding Information

The MSOE Center for BioMolecular Modeling would like to acknowledge and thank the National Institutes of Health Clinical and Translational Science Award (NIH_CTSA UL1RR031973) and the Milwaukee School of Engineering for their support in funding the 2018_2019 SMART Team program.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Multifaceted Role of Lipid A 1‐phosphatase in Bacterial Envelope Biogenesis
Authors: Zhao Jinshi , An Jinsu , Hwang Dohyeon , Wu Qinglin , Wang Su , Gillespie Robert A , Yang Eun Gyeong , Chung Hak Suke , Zhou Pei
Published: June 2021

Gram‐negative bacterial cell envelope consists of a phospholipid bilayer inner membrane, a peptidoglycan layer and an asymmetric bilayer outer membrane with lipopolysaccharide (LPS) covering the outer leaflet and phospholipid the inner leaflet. Here we report that the lipid A 1‐phosphatase (LpxE) functionally connects the biogenesis of multiple membrane components by utilizing a variety of lipid substrates. We found that the LpxE from Aquifex aeolicus (LpxEAA) is capable of dephosphorylating lipid A (the hydrophobic anchor of LPS), phosphatidyl glycerol phosphate (PGP, an intermediate in the phosphatidyl glycerol synthetic pathway), and undecaprenyl pyrophosphate (C55‐PP, an essential lipid carrier for syntheses of both peptidoglycan and O‐antigen of LPS). Overexpression of LpxEAA in E. coli complements lethal deletion strains deficient in PGP or C55‐PP synthesis. In Francisella novicida, an animal pathogen, deletion of LpxEFN leads to sensitivity to bacitracin, an antibiotic targeting C55‐PP phosphatase. In a F. novicida C55‐PP phosphatase deficient strain carrying a plasmid harboring LpxEFN, suppression of LpxE expression causes cell deformation, loss of O‐antigen and ultimately cell death. Intriguingly, LpxEAA is an efficient phosphatase to lipid A species without the 3‐Deoxy‐D‐manno‐oct‐2‐ulosonic acid (KDO) moiety. Such a unique property of LpxEAA has enabled us to engineer E. coli strains carrying an LpxEAA‐harboring plasmid to produce novel lipid A species as the major component of LPS, implicating its potential usage for biotechnology applications.

Support or Funding Information

This work was supported by the Intramural Research Program of KIST, by the Pioneer Research Center Program (2014M3C1A3054141) through the National Research Foundation of Korea funded by the Ministry of Science, ICT & Future Planning, by the National Research Foundation of Korea (NRF) grant founded by the Korea government (MSIT) (2018R1A2B2008995) and National Institutes of Health (NIH) grants (GM51310 to C.R.H.R. and P.Z. and GM115355 to P.Z.).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dietary Rice Bran Oil and γ‐Oryzanol Attenuates Toll Like Receptor‐mediated IκBα Phosphorylation, NF‐κB p65 Activation and Matrix Metallo Protease‐9 Expression in Peritoneal Macrophages from Rats Fed Partially Hydrogenated Vegetable Fat and thereby Exhibit Anti‐Inflammatory Activity
Authors: Yalagala Poorna C , Dhavamani Sugasini , Pandugala Pavan Kumar , Lokesh B.R.
Published: June 2021

In spite of warning issued by regulatory agencies all over the world, South Asian countries are continuing to use Partially Hydrogenated Vegetable Fat (PHVF) in many food products. PHVF contains trans fatty acid known as elaidic acid which is pro‐inflammatory. NF‐κB p65 and Matrix Metallo Proteases‐9 (MMP‐9) play an important role in promoting inflammation. Dietary Rice bran oil (RBO) and γ‐ORY reduce the secretion of inflammatory mediators from macrophages as compared to those obtained from rats fed diet containing PHVF and thereby exhibit anti‐inflammatory activity. The present study was focused on specific role of RBO and γ‐ORY on the phosphorylation of IκBα, subsequent activation of NF‐κB p65 and secretion of Matrix Metallo Proteases (MMP‐9) a mechanism by which RBO and γ‐ORY may be exhibiting their anti‐inflammatory activity. Male wistar rats were fed AIN‐93 diet for 60 days. The diet was supplemented with one of the following fat/oil at 10wt%level: PHVF, RBO‐N (unfrationated oil), RBO‐MCR (Minor Constituents Removed oil), RBO‐MCR+Unsaponifiable Matter from Rice Bran Oil added back to RBO‐MCR (UM RBO‐MCR), RBO‐MCR+ γ‐ORY(700mg/kg diet), PHVF+RBO‐N (5.0+5.0wt%), PHVF+ γ‐ORY (700mg/kg diet). The macrophages from rats fed diet containing PHVF showed increased phosphorylation of IκB, translocation of NF‐κB p65, enhanced secretion of MMP‐9 indicating pro‐inflammatory nature of dietary PHVF. Diet containing RBO‐N, RBO‐MCR, UM RBO‐MCR and RBO‐MCR+ γ‐ORY significantly reduced phosphorylation of IκB, translocation of NF‐κB p65 and expression of MMP‐9 in macrophages all of which were enhanced when PHVF diets were fed to rats. Thus RBO and its components like γ‐ORY exhibit anti‐inflammatory activity by modulating expression of NF‐κB p65 which ultimately results in decreased secretion of pro‐inflammatory mediators by macrophages.

Support or Funding Information

Department of Biotechnology, India

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PRECLINIC EVALUATION OF SIRNAS AGAINST VEGFR‐2 FOR THE TREATMENT OF DIABETIC RETINOPATHY
Authors: Juarez Gerardo Vera , Villafaña Santiago
Published: June 2021

The vascular endothelial growth factor (VEGF) is a group of important proteins in the signaling for vasculogenesis and angiogenesis, it has been reported to participate in the development of diabetic retinopathy (RD), which is the second cause of blindness worldwide, within the VEFG pathway, it has been reported that VEGFR‐2 is the principal involved in RD, for this reason, the aim of this work is to evaluate the expression of VEFGR‐2 in the retina of diabetic rats and also design and synthesize a siRNA to the VEFGR‐2. Wistar rats were inyected with streptozotocin (60mg/kg/i.p), after 4, 8 and 12 weeks the neovascularization was evaluated by junctions and lacunarity, the retina was extracted and the expression of VEFGR‐2 was determined by RT‐PCR. For the siRNA's design, we used Wizard v3.1, RNAfold and EMBOSS software programs, the selected sequences were synthesized with a MERMADE 8 and evaluated. The results obtained showed that there is an increase of junctions and in the levels of VEGFR‐2 and a decrease of lacunarity in the retina with 8 and 12 weeks of diabetes evolution.

The evaluation of siRNA suggests that 343–364 and 2621–2641 regions are susceptible to degrade. Preliminary results showed a decrease in the neovascularization when we administrated the siRNAs. Thus, our conclusions are that early diabetes can produce DR and our siRNA maybe could decrease the DR evolution.

Support or Funding Information

Grants: Instituto Politécnico Nacional (SIP‐20194983), Programa Institucional de Formación de Investigadores del IPN (BEIFI No. 2062), Scholarship by Consejo Nacional de Ciencia y Tecnología (CONACYT No.630882) and Comisión de Operación y Fomento de Actividades del IPN (COFAA).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Post Stroke Depression: the Mechanisms, Prevention and Therapy Through Holistic Treatment Approach
Authors: Santos Maria
Published: June 2021

Background/Objectives

Stroke is the third most common cause of disability and second most common cause of death worldwide. At least one‐third of stroke survivors suffer from depression.

Depression is a common complication of stroke, leading to increased morbidity and mortality, and is defined as depressed mood on a daily basis for a minimum duration of two weeks.

Objective is to investigate the causal associations between post stroke depression and its outcomes. To evaluate the associations between stroke and depression in Nigerians adults living American and in Nigeria

Method

Investigation of post stroke depression in American adults and Nigerian‐American adult living in America and Nigeria.

Holistic clinical approach interventions when treating PSD for better outcome than pharmacological only approach. These includes: Adult day care, dietitian, speech therapy, social services, PT, OT, yoga, psycho‐therapy, primary care clinicians, family and friends.

Results/discussion

Study participants living with other people did better than those living alone P‐value 0.05.

Conclusion

Stroke has been identified as one of the prioritized non‐communicable diseases the hope is that the promotion and endorsement of new, more‐effective approaches for the prevention of stroke and CVD, in combination with prevention strategies, will change current practice worldwide, thereby saving millions of lives. By complementing cost‐effective individual health‐care interventions with population‐wide prevention strategies, to make a substantial impact on the global non‐communicable diseases. There are many factors that have effects on PSD. The severity of stroke is an important factor in the occurrence of PSD. Mental history is a possible predictor of PSD. Prevention of PSD requires social and family participation, and holistic therapy approach.

RECOMMENDATIONS

The recommendations listed below must be considered.

For any successful outcome in treating PSD, nursing, social worker, family members, and power of attorney or guardians should team up for better living arrangement, because this study showed that patients living with others have better outcomes than those living alone.

Social determinant factors cannot be ignored in this patient population. Considering social determinant factors such as level of poverty, less than high school education, median house income, employment, transportation factors all play a role for successful outcome in managing PSD.

Support or Funding Information

Einstein Institute ‐ USAT

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of Actionable Genomic Alterations in Tissue Using Idylla™
Authors: McKillip Kelsey Dillehay , Koch Matthew , Hodges Kurt
Published: June 2021

Quick and reliable identification of actionable genomic alterations is essential to providing high‐quality cancer care. Herein we describe our assessment of Idylla™, a fully automated, cartridge‐based, PCR system for the qualitative detection of actionable genomic alterations in formalin‐fixed, paraffin‐embedded (FFPE) tissue. Three different Idylla™ assays were evaluated to demonstrate the system's ease of use, concordance with established methodology, and overall clinical utility.

Thirty‐six patient samples, previously characterized using a clinically validated method, were assessed for EGFR mutations, BRAF mutations, or microsatellite instability (MSI) using Idylla™. The Idylla™ MSI Mutation Assay, which detects mutations in seven novel, tumor specific MSI loci (ACVR2A, BTBD7, DIDO1, MRE11, RYR3, SEC31A, and SULF2), was used to assess microsatellite stability in twelve cases of colorectal cancer. The Idylla™ EFGR Mutation Assay, which detects 51 mutations in exons 18, 19, 20, and 21 of the EGFR gene, was used to identify mutations in twelve cases of lung cancer. Lastly, the Idylla™ BRAF Mutation Assay, which detects seven mutations in codon 600 of the BRAF oncogene, was used to detect mutations in twelve cases of melanoma.

Briefly, one 10μm FFPE tissue section was loaded into the cartridge per the manufacturer's instructions. No manual macrodissection was performed. A representative H&E stained slide was retrospectively reviewed and scored for neoplastic cell content by a Pathologist (K.H.). Idylla™ assay results were compared to previously reported results. Three cases, one per assay, were excluded from the final analysis due to lack of neoplastic cell content (NT = no tumor) or assay incompatibility.

Overall, the adjusted concordance between the Idylla™ assays and clinically validated methodologies was high. The Idylla™ MSI Mutation Assay was 100% (11/11) concordant with the immunohistochemical assay for mismatch repair (MMR), which included markers MLH1, MSH2, MSH6, and PMS2. The Idylla™ EGFR and Idylla™ BRAF Mutation assays were 91% (10/11) concordant with Sanger sequencing results. Additional studies utilizing a third methodology are needed to further evaluate discordant results.

The Idylla™ assays showed high concordance with previously reported results generated using clinically validated methods. Advantages of the Idylla™ system include minimal hands‐on time and rapid turnaround time. In conclusion, Idylla™ offers fast and reliable analysis for a variety of actionable genomic alterations in FFPE tissue.

Support or Funding Information

Support for this project was provided by the Department of Pathology and Laboratory Medicine.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A New Series of Gs‐ and β‐Arrestin‐biased β2‐Adrenoceptor Agonists
Authors: Woo Anthony Yiu‐Ho , Ge Xinyue , Pan Li , Xing Gang , Mo Yongmei , Xing Ruijuan , Li Xiaoran , Zhang Yuyang , Wainer Irving W. , Cheng Maosheng , Xiao Rui‐Ping
Published: June 2021

β‐Arrestins are implicated in desensitization of activated G protein‐coupled receptors (GPCRs) and transduction of signals in their own right. Biased ligands possessing different efficacies in activating different G protein‐dependent and β‐arrestin‐dependent signals downstream of a single GPCR have been described. Here, we report the characterization of agonist bias for compounds developed along a drug discovery project of β2‐adrenoceptor agonists. Compounds, including derivatives of fenoterol, 2‐amino‐1‐phenylethanol and 2‐amino‐2‐phenylethanol, were obtained or synthesized and their effects on cardiomyocyte contractility, cAMP synthesis and β‐arrestin recruitment were determined in β2‐adrenoceptor‐expressing cells. Gs versus β‐arrestin bias were analyzed by the operational model. Gs versus Gs/Gi bias were studied by means of sensitivity towards pertussis toxin, a Gi disruptor. Three compounds with a core structure of 5‐(1‐amino‐2‐hydroxyethyl)‐8‐hydroxyquinolin‐2(1H)‐one were identified as β‐arrestin‐biased β2‐adrenoceptor agonists. These findings would facilitate the development of novel drugs for the treatment of heart failure and asthma.

Support or Funding Information

This work was supported by the Ministry of Science and Technology of the People's Republic of China under the National Key Research and Development Program of China [2018YFA0507603], the National Science and Technology Major Project [2013ZX09507001‐001002, 2013ZX09301305‐001, 2013ZX09508104, 2018ZX09739009], the National Basic Research Program [2012CB518000], the National Natural Science Foundation of China [81673355, 81872752, 31521062, 81630008, 81790621], Beijing Municipal Science & Technology Commission [Z171100000417006], and the intramural research program of the NIH, USA.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Osteogenic Differentiation of Normotensive Rats Depends on the β‐adrenergic Receptor
Authors: Barreto Ayná Emanuelli Alves , Brito Victor Gustavo Balera , Ballassoni Beatriz Babeto , Frasnelli Sabrina Tfaile , Patrocinio Mariana Sousa , Santos Carlos Ferreira , Lara Vanessa Soares , de Oliveira Sandra Helena Penha
Published: June 2021

Bone remodeling is a complex process that depends on the balance between formation and resorption bone, a process which is regulated by bone cells and systemic factors, like the Sympathetic Nervous system (SNS). The mediators of these system are able to regulate bone metabolism through adrenergic receptors on the surface of the osteoblasts. However, the role of β‐adrenergic receptors is not clear in the osteogenic differentiation process. Thus, in this study we aimed to evaluate the role of B‐adrenergic receptor on osteoblastic differentiation of bone marrow mesenchymal stem cells from normotensive and Spontaneously Hypertensive Rats (SHR).

Methods

70‐days‐old male Wistar and SHR rats were used for bone marrow collection from femurs, which was placed in cell culture flasks and after in to 24‐well plates, where they received osteogenic medium (OM: MEM, plus 50 μg/mL ascorbic acid, 10 mM β glycerophosphate, and 10−8 M dexamethasone) and the treatment with Carvedilol (1μM), non‐selective adrenergic receptor antagonist, or Nebivolol (0,1 μM), β1‐adrenergic receptor antagonist. Cell proliferation (MTT assay) and alkaline phosphatase specific activity (Alp) were analyzed at day 7, 10 and 14. Mineralization was evaluated at day 14, by Alizarin Red S. Gene expression of osteogenic markers and B1 and B2‐adrenergic receptor were evaluated at day 7, by real time‐RT‐PCR. The proteolytic activity of matrix metalloproteinase 2 and 9 (MMP‐2 and MMP‐9) were evaluated at day 7 using gelatin zymography. The protocol was approved by Institutional Animal Care and Use Committees (School of Dentistry of Araçatuba; Process 00686‐2016).

Results

Alp activity increased in OM group from Wistar rats at day 7 and the Nebivolol group reduced those activity at day 7 and at day 10. SHR OM group did not present Alp increase, and the treatment with both drugs did not change it. Nebivolol treatment decreased Runx2, Osterix and β‐catenin in Wistar compared to OM. In SHR, only Osterix was reduced. Alp, Osteocalcin and Bone sialoprotein have significative decrease in cells from Wistar treated with Nebivolol compared to OM. Pro and active MMP‐2 was reduced by Nebivolol treatment just at Wistar cells. Besides that, Nebivolol reduced Adrb1 gene expression at day 7 in Wistar group compared to OM. Mineralization showed that Nebivolol reduced mineral deposition in Wistar compared to OM.

Conclusion

β1 adrenergic receptor seems to be involved in the osteogenic differentiation of cells from Wistar rats.

Support or Funding Information

FAPESP (Grant: #2015/03965‐2) and CAPES.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mast Cell Supernatant Inhibit Bone Formation Markers Gene Expression
Authors: Frasnelli Sabrina Cruz Tfaile , Jordão José VR , Linjardi Maria Carolina , Balassoni Beatriz B , Brito Victor Gustavo B , Barreto Ayna E A , Lara Vanessa S , Santos Carlos F , Oliveira Sandra HP
Published: June 2021

Spontaneously hypertensive rats (SHR) present genetic alterations in bone metabolism and in adaptive immunity. Mast cells (MCs) play a fundamental role in inflammatory response, tissue repair and defense of the organism and may also be involved in the bone metabolism. Thus, in this study we aimed to evaluate the influence of mast cell mediators in the acquisition of the osteogenic phenotype in vitro and in the gene expression of bone markers from calvariae cells of SHR.

Methods

Primary culture from neonatal calvariae obtained by explant technique were expanded and plated in 24‐well culture plates where they received osteogenic medium (OM, ascorbic acid, β‐glycerophosphate and dexamethasone). The RBL‐2H3 MC lines were adapted to osteogenic medium, subsequently stimulated via FcɛRI, and culture supernatant was collected and was added directly into the calvariae culture in concentration of 15%, 10% and 5%. Cell viability was accessed by MTT assay, and gene expression of osteogenic markers was evaluated at day 7 by qRT‐PCR. The protocol was approved by Institutional Animal Care and Use Committees (School of Dentistry of Araçatuba; Process:00490‐2018).

Results

MTT assay did not show a cytotoxic potential from MC supernatant on calvariae culture. MC supernatant, in all concentration, significantly decreased gene expression of the transcription factors Runx2, Osterix, but not β‐catenin. Moreover, it decreased osteocalcin and bone alkaline phosphatase expression, in a dose‐dependent manner. Bone sialoprotein expression was decreased in response to the higher MC supernatant concentration, but the opposite was observed in response to the lower concentration. Osteopontin and type 1 collagen expression were decreased, in a similar manner by all MC supernatant concentrations.

Conclusion

Our current data allow us to suggest that MC‐derived factors can directly inhibit bone formation‐related markers expression. Further studies are being performed to better understand the mechanisms related to mast cell – bone cell crosstalk.

Support or Funding Information

Financial support and acknowledgments: FAPESP (Grant: #2015/03965‐2 and Grant: #2017/05873‐3)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Autophagy in Cardiomyocytes by Calmodulin Antagonists
Authors: Lopez Vanessa , Hayden Matt , Giles Jennifer , Keomanivong Faithe , Wauson Eric M , Tran Quang‐Kim
Published: June 2021

Macroautophagy (hereafter referred to as autophagy) is a catabolic process that regulates important aspects of myocardial metabolism. While numerous studies have indicated involvement of signaling via Ca2+ in autophagic flux, the precise mechanisms remain unknown. Calmodulin (CaM) is a ubiquitous transducer of intracellular Ca2+ signals by interacting with an estimated network of 300 cellular proteins. We have begun to examine the role of CaM in autophagic flux in cardiomyocytes. A basal level of autophagy is required for normal myocardial function. Chloroquine dose‐dependently reduces basal autophagic flux in H9c2 cardiomyocytes, as indicated by LC3 immunoblotting, p62 immunofluorescence, and autophagic flux assays. Competitive binding assays using purified CaM and FRET‐based CaM biosensors demonstrate direct Ca2+‐dependent CaM binding to chloroquine. The structurally distinct CaM antagonists W‐7, trifluoperazine, and CGS 9343B also dose‐dependently inhibit basal autophagic flux by reducing autophagosome degradation. Most CaM‐dependent processes depend at the front end on the amplitude and dynamics of intracellular Ca2+ signals. Interestingly, chloroquine, W‐7, trifluoperazine, and CGS 9343b all mobilize intracellular Ca2+. The effects of these pharmacological agents to inhibit autophagic flux are thus associated with their inhibition of CaM and not of initial Ca2+ signals produced in cells. These data indicate a role for CaM in the control of autophagic flux in cardiomyocytes.

Support or Funding Information

American Heart Association (15SDG25090279) and Iowa Osteopathic Education and Research Funds (Eric Wauson); NIH HL112184 and DMU IOER‐090717 (Quang‐Kim Tran)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Novel Dopamine D4 Receptor‐Selective Compounds Reveal Structure‐Activity Relationships that Engender Agonist Efficacy
Authors: Keck Thomas M , Free R. Benjamin , Day Marilyn M , Fountain Griffin , Cooper Charles , Bonifazi Alessandro , Newman Amy Hauck , Sibley David R , Wu Chun , Boateng Comfort
Published: June 2021

The dopamine D4 receptor (D4R) plays important roles in cognition, attention, and decision making. Novel D4R‐selective ligands have promise in medication development for neuropsychiatric conditions, including Alzheimer's disease and substance use disorders. To identify new D4R‐selective ligands, and to understand the molecular determinants of agonist efficacy at D4R, we report a series of eighteen novel ligands based on the classical D4R agonist A‐412997 (2‐(4‐(pyridin‐2‐yl)piperidin‐1‐yl)‐N‐(m‐tolyl)acetamide). We report a series of eighteen novel D4R ligands with varied substitutions on the phenylpiperidinyl ring, linker region, and/or the arylamide moieties. Compounds were profiled using radioligand binding displacement assays, β‐arrestin recruitment assays, cAMP inhibition assays, and molecular dynamics computational modeling. We identified several novel high‐affinity (Ki ≤ 4.3 nM) and D4R‐selective (>100‐fold vs other D2‐like receptors) compounds with diverse partial agonist and antagonist profiles, falling into three structural groups. Group 1 modifications resulted in a gain of efficacy at D2Rs and D3Rs. Group 2 modifications resulted in lower partial agonist efficacy at D4R. Group 3 modifications resulted in high‐potency full antagonists at D4R. These compounds inform receptor‐ligand interactions that control efficacy at D2‐like receptors and may provide insights to targeted drug discovery leading to new compound development and new tools for a better understanding of the role of D4Rs in neuropsychiatric disorders.

Support or Funding Information

Support for this research was provided by High Point University, Fred Wilson School of Pharmacy. Additional support was provided by the National Institute of Neurological Disorders and Stroke‐Intramural Research Program, National Institute on Drug Abuse‐Intramural Research Program (Z1A DA000609), and Rowan University.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sporoderm‐broken spores of Ganoderma lucidum water extract (BSGLWE) inhibits high‐fat diet induced obesity through regulating gut microbiota in mice
Authors: Guo Chenjie , Sang Tingting , Guo Dandan , Li Mingyan , Li Zhenhao , Wu Kaikai , Xu Jing , Na Kun , Wang Xingya
Published: June 2021

Obesity is a worldwide epidemic disease associated with low‐grade chronic inflammation. Recent evidence shows that intestinal dysbiosis contributes to obesity and its comorbidities. Ganoderma lucidum (G. lucidum) is a well‐known medicinal mushroom in traditional Chinese medicine with antidiabetic effect. In this study, we examined whether sporoderm‐broken spores of Ganoderma lucidum water extract (BSGLWE), mainly contains G. lucidum polysaccharides (GLPs), could regulate gut microbiota and inhibit obesity and insulin resistance. BSGLWE at different concentrations (100 mg/kg and 300 mg/kg) were individually administered to HFD‐fed mice and LFD‐fed mice for 12 weeks. We found that BSGLWE inhibit high‐fat diet (HFD) induced weight gain, visceral fat accumulation, inflammation in HFD‐fed mice. BSGLWE decreased the HFD‐induced high level of blood lipid, especially free fatty acids (FFAs) and low‐density lipoprotein (LDL) in HFD‐fed mice. In addition, BSGLWE inhibited TLR4/Myd88 signal pathway in white adipose tissues. 16S rDNA sequencing demonstrated that BSGLWE treatment significantly changed the composition of the gut microbiota in mice fed HFD. Gene expression analysis showed down‐regulation of pro‐inflammatory cytokines (TNF‐α, IL‐6, IL‐1β) in tissues and up‐regulation of tight junction proteins (occludin and ZO‐1), Muc2 and Reg3g in small intestine after BSGLWE treatment. Moreover, BSGLWE also reversed HFD‐induced gut dysbiosis and improved the intestinal barrier functions in the HFD‐fed mice. In conclusion, our date show that the BSGLWE might be used as prebiotic agents to prevent obesity‐related metabolic disorders.

Support or Funding Information

Supported by Science and Technology Planning Project of Zhejiang Province (Grant No. 2019C02100) and National Natural Science Foundation of China (Grant No. 81473397).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Physiology Understanding (PhUn) Week at Texas A&M University‐Kingsville: a Retrospective Study
Authors: Lopez Hector , Velez‐Hernandez Maria E
Published: June 2021

Texas A&M University‐Kingsville (TAMUK) is located in Kingsville, Texas and serves a highly diverse student body of around 9,000 students. PhUn week is a K‐12 community outreach activity that in 2014 had 86 events nationwide (www.aps.org). Out of the 86 events, only three of them were in Texas, and none of them south of San Antonio. During the Fall semesters of 2016, 2017 and 2018 the Biological and Health Sciences (BHS) Department at TAMUK offered the course Understanding Human Physiology (BIOL 4355) centered on service learning. As part of this course, students developed ways to teach newly acquired physiology concepts to elementary and middle school students. Interactive workshops were presented during Physiology Understanding Week as a way to showcase what students had learned in the areas of cardiovascular, immune, respiratory, renal and nervous system. Support from the American Physiological Society, Service Learning initiatives at TAMUK, and the TAMUK BHS Department were critical for the success of this program. Here we will discuss how in 3 years, a total of 41 TAMUK undergraduate presenters and 13 volunteers impacted five schools and more than 500 elementary and middle school students. PhUn week activities allowed the university to build local partnerships between science teachers and student scientists. In addition, this kind of engaging activities served as a platform for students to reach out to our communities, and to motivate the next generation of scientists.

Support or Funding Information

This project was funded by The American Physiological Society, Texas A&M Service Learning Initiative and the TAMUK Biological and Health Sciences Department.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Susceptibility of erythrocytes from different sources to xenobiotics‐induced lysis
Authors: crupi rosalia , Morabito Rossana , Remigante Alessia , Gugliandolo Enrico , Britti Domenico , Cuzzocrea Salvatore , Marino Angela
Published: June 2021

As erythrocytes are continuously exposed to plenty of metabolites and toxicants, the aim of the present work is to show whether erythrocytes from different sources may exhibit different susceptibility to hemolysis induced by two classes of xenobiotics, Cnidaria venoms, acting via pore forming mechanism on cell membrane, and Cd2+, inducing cell damage mainly via lipid peroxidation. To this end, the hemolytic power of crude venom from stinging cells of the anthozoan Aiptasia mutabilis and from the scyphozoan Pelagia noctiluca has been tested, along with Cd2+ (5 to 20 mM), on erythrocytes (0.05% v/v) obtained from either rabbit, or dog, or chicken, or human source. Our results show a more significant susceptibility of rabbit erythrocytes to hemolysis induced by both crude venom and Cd2+ than erythrocytes from other sources. This difference seems to rely neither on the different specimens used to extract crude venom, nor on the different mechanism of toxicants. In this light, the present study may contribute: i) to confirm hemolytic test as a suitable biological assay to verify erythrocytes resistance to toxicants; ii) to show variability in hemolytic response to xenobiotics; iii) to propose rabbit erythrocytes as more sensitive to the lytic action of xenobiotics, adding more knowledge about clinical practice and drug development in these animals.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Examining the effects of housing temperature on mitochondrial adaptations to exercise and the browning of white adipose tissue
Authors: McKie Greg Lawrence , Medak Kyle D. , Knuth Carly M. , Townsend Logan K. , Shamshoum Hesham , Peppler Willem T. , Wright David C.
Published: June 2021

The thermoneutral zone for small rodents is ~29–31°C and with decreases in ambient temperature below this there are consequent increases in metabolism to promote thermogenesis. Despite this, many studies report housing mice at room temperature for the convenience of the researchers studying them. As such, the conflicting reports between human and rodent models of exercise‐induced mitochondrial biogenesis in adipose tissue and the browning of white adipose tissue may be explained by the often‐overlooked variable, housing temperature. Male 8‐week‐old C57BL/6 mice were housed in cages with (VWR) or without (SED) running wheels (n = 12/group/condition) for 5–6 weeks at room temperature (RT; 22°C) or at thermoneutrality (TN; ~29°C). Mice housed at TN ran a greater distance. VWR protected against weight gain independent of housing temperature however food intake was greater in both VWR and SED mice housed at RT compared to TN. Inguinal white adipose tissue (iWAT) and epididymal white adipose tissue (eWAT) mass were reduced following VWR independent of housing temperature and there were main effects of both exercise and temperature to reduce brown adipose tissue (BAT) mass. There was no effect of housing temperature on circulating glucose or glycerol concentrations however NEFA, insulin, and the HOMA‐IR were increased at TN. In eWAT there was a main effect of exercise to increase cytochrome c oxidoreductase core I subunit (CORE1), cytochrome c oxidase, and peroxisome proliferator‐activated receptor‐γ coactivator‐1α (PGC‐1α) protein content with a main effect of housing temperature for PGC‐1α as well. There was a strong trend for an exercise‐induced increase in pyruvate dehydrogenase (PDH). In iWAT, there was a main effect of exercise and temperature on mitochondrial proteins with VWR leading to the expected increases in PDH, CORE1, COX IV, and PGC‐1α, but reduced markers of mitochondrial content at TN. Most notably protein content of the thermogenic uncoupling protein‐1 (UCP‐1) was undetectable in iWAT at TN. Qualitative histological analysis revealed patches of multilocular adipocytes in iWAT with VWR at RT but not at TN. Together our findings provide novel evidence that while housing mice at TN reduces markers of mitochondrial content in adipose tissue under sedentary conditions, the ability of exercise to increase mitochondrial protein content remains largely intact.

Support or Funding Information

Ontario Graduate Scholarship Canada Research Chairs Program

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Muscle contractile activity‐mediated regulation of antioxidant enzymes in oxidative muscle requires p62/SQSTM1 phosphorylation‐induced Nrf2 activation
Authors: Yamada Mami , Iwata Masahiro , Warabi Eiji , Oishi Hisashi , Lira Vitor A , Okutsu Mitsuharu
Published: June 2021

Enhanced muscle contractile activity, as observed with regular exercise, prevents oxidative stress‐induced muscle wasting, at least partially by improving the antioxidant defense system. p62/sequestosome1 (p62/SQSTM1) competitively binds to the kelch‐like ECH‐associated protein 1, activating nuclear factor erythroid 2‐related factor 2 (Nrf2), which stimulates transcription of genes containing antioxidant/electrophile responsive elements. However, it remains to be determined if this process is activated by regular exercise in skeletal muscle. Here we demonstrate that increased muscle contractile activity enhances antioxidant enzyme expression, Nrf2 translocation into nuclei, and Nrf2 DNA‐binding activity in mouse skeletal muscle. Basal skeletal muscle expression of the Nrf2 target antioxidant gene NAD(P)H‐quinone oxidoreductase 1 (NQO1) is markedly reduced in muscle‐specific Nrf2 knockout (Nrf2 mKO) mice and muscle‐specific p62 knockout (p62 mKO) mice when compared to wild type littermates (WT). Using voluntary running, we demonstrate that regular endurance exercise increases levels of total p62 and Ser 351 phosphorylation of p62, an event that enhances p62 affinity to Keap1, in the oxidative soleus muscle. Regular exercise fails to increase the expression of the antioxidant proteins CuZnSOD (i.e., SOD1) and EcSOD (i.e., SOD3) in the soleus of Nrf2 mKO and p62 mKO. However, the exercise‐mediated elevation of these proteins is preserved in glycolytic skeletal muscle of Nrf2 mKO and p62 mKO mice. Collectively, these findings indicate that p62 and Nrf2 cooperatively regulate the exercise‐mediated increase of antioxidant enzyme expression in oxidative muscle, and that an alternate molecular regulation of these genes predominates in glycolytic muscle fibers in response to regular exercise.

Support or Funding Information

This study was supported by Grant‐in‐Aid for Research Activity start‐up (25882036), Grant‐in‐Aid for Scientific Research (B) (15H03080), Grant‐in‐Aid for Scientific Research (B)(18H03153), Suzuken Memorial Foundation, Toyoaki Scholarship Foundation, The Nakatomi Foundation, and The Uehara Memorial Foundation to Mitsuharu Okutsu.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.