bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ganoderma Lucidum polysaccharide inhibits lipid accumulation in vivo and in vitro by activating HIF‐1a through ERK1/2 signaling pathway
Authors: Wang Ying , Guo Chenjie , Pan Haitao , Sang Tingting , Wang Yujie , Guo Dandan , Wang Xingya 
Published: June 2021

Ganoderma lucidum polysaccharide (GLP) has been proposed to have beneficial effects against obesity. However, how GLP affects lipid accumulation has not been examined. In this study, we used 3T3‐L1 preadipocytes and a high‐fat diet‐induced obese mice model to investigate the effects and mechanism of GLP on lipid accumulation. Our results revealed that GLP significantly inhibited adipogenic differentiation in a dose‐dependent manner in 3T3‐L1 preadipocytes determined by Oil Red O staining and Triglyceride measurement. In order to explore the mechanism, we conducted a dynamic transcriptome study during 3T3‐L1 preadipocytes differentiation (0 h, 9 h, 18 h, 48 h, 120 h) with or without GLP treatment. Transcriptome analyses showed that GLP significantly caused cellular hypoxia by blocking respiration chain, and inducing the expression of HIF‐1a which is a major regulator of cell cycle arrest during hypoxia by activating the ERK1/2 signaling pathway. And then delayed the mitotic clonal expansion(MCE) which is required for differentiation of 3T3‐L1 cells, eventually inhibiting a cascade of adipogenesis pathway. These has been verified by qRT‐PCR or Western blot. Moreover, flow cytometry data revealed that GLP does arrest cell cycle at G0/G1 phase at 9 h of differentiation. Furthermore, knocking‐down of HIF‐1a by siRNA can abolish the GLP‐induced inhibition of adipogenesis in 3T3‐L1 cells, indicating that HIF‐1a was directly required for GLP‐induced inhibition of aidopogenesis. In addition, we demonstrated that GLP induces the phosphorylation of ERK1/2. Interestingly, pretreatment with ERK inhibitor, significantly rescued GLP‐induced inhibition of adipogenesis. We further demonstrated that HIF‐1a is the downstream target of ERK1/2. Furthermore, GLP treatment significantly suppressed hepatic lipid accumulation by activating ERK1/2/HIF‐1a signaling pathway to improve non‐alcoholic fatty liver disease(NAFLD) and insulin resistance in high‐fat diet‐induced obese mice. Taken together, GLP inhibits lipid accumulation by activating HIF‐1a through ERK1/2 signaling pathway in vivo and in vitro. These findings also indicate that GLP possesses protective effects in NAFLD and IR‐related metabolic disorders.Support or Funding InformationNational 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.
Distinct Missense Mutations of the Parkinson's Disease‐Related Ubiquitin Kinase PINK1 Alter Auto‐ or Substrate‐Phosphorylation
Authors: Springer Wolfdieter , Truban Dominika , Caulfield Thomas R , Hudec Roman , Hou Xu , Lewis‐Tuffin Laura J , McCarty Arthur M , Soto Alexandra I , Walton Ronald L , Puschmann Andreas , Wszolek Zbigniew K , Ross Owen A , Fiesel Fabienne C 
Published: June 2021

The mitochondrial ubiquitin (Ub) kinase PINK1 and the cytosolic E3 Ub ligase PRKN (Parkin) together mediate the selective degradation of damaged mitochondria via the autophagy‐lysosome system (mitophagy). Complete loss of either gene function results in failure of this mitochondrial quality control and thus accumulation of damaged organelles. While homozygous or compound heterozygous mutations in PINK1 or PRKN are the most common causes of early‐onset Parkinson's disease (PD), the contribution of heterozygous variants to PD is debated. However, recently we identified a partial dominant‐negative effect of a heterozygous PINK1 missense mutations that is associated with a significant increased risk and an earlier onset compared to late‐onset, sporadic PD. This underscores that not all missense mutations are alike and suggests critical threshold functions of this stress‐activated, neuroprotective pathway.Here, we analyzed the underlying mechanisms in greater detail and found that the pathogenic PINK1 mutant, which can dimerize with wild‐type PINK1 protein, is aberrantly auto‐phosphorylated, impacting its enzymatic activity towards the substrates Ub and PRKN. Phosphorylation of both PINK1 targets however, is critical for PRKN recruitment as well as efficient labeling of mitochondria with phosphorylated poly‐Ub chains that serve as the mitophagy tag. In addition to cells from PD patients, we confirmed the altered phosphorylation pattern of mutant PINK1 under endogenous conditions in isogenic CRISPR/Cas9 genome engineered cells. Excitingly, while studying the pathogenic mechanisms in detail with a structure‐function approach, we identified a novel PINK1 variant that exerts even greater kinase activity than wild type PINK1. Our results further suggest that this hyperactive form of PINK1 likely modulates substrate receptivity thereby resulting in enhanced kinase activity and greater mitophagy rates. Going forward, it will be interesting to study if enhanced PINK1 signaling translates into greater neuroprotection. An in‐depth understanding of the regulation and enzyme functions of PINK1 may provide a rationale for a structure‐function based drug design in the future.Support or Funding InformationThis work was partially supported by funding from the Mayo Clinic Foundation, the National Institutes of Health (NIH)/National Institute of Neurological Disorders and Stroke (NINDS) [R01#NS085070] and the Michael J. Fox Foundation for Parkinson's Research to W.S.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Designing and Synthesis of a siRNA against Angiopoietin‐2 for the Treatment of Neovascularization Associated to Diabetic Retinopathy
Authors: Becerra Sandra Edith Cabrera , Villafaña Santiago 
Published: June 2021

Diabetic retinopathy (RD) is a pathology that causes changes in the retina, for example exudates, microhemorrhages, neovascularizations, etc., which leads to loss of sight and is the third cause of blindness in the world. Among the mechanisms involved in neovascularization are angiogenic factors like angiopoietins (Angpt), especially angiopoietin‐2 (Angpt2), which has been reported that increase the formation of new vessels. For this reason, the aim of this study is to design and synthesize an siRNA against to Angpt2. Male Wistar rats were administered with streptozotocin (STZ) to induce diabetes, after 4 weeks the biochemical parameters, the neovascularization (junctions and lacunarity) and the expression of the Angpt2 were determined in the retina. The siRNA was designed with the siRNA Wizard v3.1 and RNA fold webserver software programs and was synthesized by MERMADE 8 equipment. The results showed that the number of junctions and the levels of Angpt2 of the diabetic rats were higher than the controls significantly, while the lacunarity was significantly lower than controls, on the other hand, the region susceptible to silencing could be from 1416 to 1436. Thus, we can conclude that early diabetes produces an increase in Angpt2 and neovascularization, which could be reduced by the siRNA to Angpt2.Support or Funding InformationGrants: Instituto Politécnico Nacional (SIP‐20194983), Programa Institucional de Formación de Investigadores del IPN (BEIFI No. 2177), Scholarship by Consejo Nacional de Ciencia y Tecnología (CONACYT No.418913) 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.
Oleocanthal as a Promising Natural Therapeutic Agent: the Experimental Leishmaniasis Paradigm
Authors: DOTSIKA ELENI , KARAMPETSOU KALLIOPI , KOUTSONI OLGA S. , ANGELIS APOSTOLOS , CHALAMBALAKI MARIA , SKALTSOUNIS LEANDROS‐ALEXIOS 
Published: June 2021

Oleocanthal (OC) is a phenolic compound of Extra Virgin Olive Oil (EVOO), the main source of fatty acids in the Mediterranean diet that is associated with beneficial health effects such as reduced incidence of cardiovascular mortality and cancer. OC is responsible for the localized irritation in the oropharangeal region, is homologous with the non‐steroidal anti‐inflammatory drug ibuprofen and several studies evidenced relevant pharmacological properties of OC in different pathogenic processes. Our group has previously elucidated the anti‐leishmanial effect of Total Phenolic Fraction derived from EVOO and of the phenolic compound Oleuropein. Current study aimed to evaluate the effect of OC in murine experimental models of cutaneous and visceral leishmaniasis since there is no effective vaccine and the chemotherapeutic treatments have serious drawbacks such as toxicity, parasite resistance and high cost. BALB/c mice were infected either subcutaneously in the footpad with 106 L. major promastigotes (Cutaneous Model ‐ CM) or intravenously in the tail vein with 107 L. infantum promastigotes (Visceral Model ‐ VM), according to EC regulation conformed by FASEB Statement of Principles for the use of Experimental Animals. At 7 days post infection, mice were intraperitonealy treated with 5 and 10 mg/kg body weight of OC, 3 days per week for 5 weeks. Mice of the positive control group received orally 15 mg/kg b.w of the reference drug miltefosine (HePC), for 25 days. In the CM, results demonstrate significant reduction of the footpad swelling in OC‐treated mice along with the reduction of parasite load in the popliteal lymph nodes, 1 month post‐treatment termination. Correspondingly, in the VM, OC promoted a severe reduction of parasite burden in target organs, liver and spleen at 10 days post‐treatment termination. Moreover, in order to validate the profile of cytokines induced by OC treatment, intracellular cytokine production in splenocytes of all experimental groups was determined for IL‐4 (Th2‐type cytokine) and IL‐12 (Th1‐type cytokine) at 48h post Soluble Leishmania Antigen in vitro re‐stimulation and revealed that OC was able to stimulate important reduction in IL‐4 production by CD4+ T cells in both CM and VM, while the IL‐12 production was equal in all experimental groups. Furthermore, specific antibodies of both IgG1 and IgG2α isotypes, were detected in the plasma of OC‐treated mice. These data indicate that OC treatment resulted in the induction of a mixed Th1/Th2 immune response moderately polarized towards Th1 type that effectively mediated the elimination of parasitic spread in both disease models. Therefore, OC appears as a potential therapeutic agent into the leishmaniasis drug discovery pipeline.Support or Funding InformationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Airway Caliber is Linked to Wall Thickness in Mice: Implications for Morphometry
Authors: Meyerholz David K , Beck Amanda P 
Published: June 2021

Morphometry of lung tissues often involves evaluation for changes in airway wall components such as collagen and/or smooth muscle. Alterations of these tissue parameters may be markers of diseases like asthma, chronic obstructive pulmonary disease or cystic fibrosis. The mouse lung is composed of airways that exhibit monopodial branching during development and these intrapulmonary airways are exclusively bronchioles (i.e. no intra‐pulmonary bronchi). As such, many airways can look alike for analysis by inexperienced observers. We tested if the airway caliber in wild type B6 mice could influence the detection and scoring of airway collagen or smooth muscle composition. We examined 15 lungs from archival blocks that came from studies with appropriate institutional approvals. Airway lumen diameter as well as wall thickness (sum of epithelium, smooth muscle and collagen) measurements of the 2 largest and 2 smallest airways were taken. These were then cumulatively analyzed for wall thickness (y axis) and airway lumen diameter (x axis). There was a significant positive slope (P=0.0017). We then grouped the data into large (lumen >130 microns) and small (lumen <130 microns) airways and compared the wall thickness, which was significantly thicker in the larger airways (P<0.0001). These data indicate that morphometry of murine airways requires analysis of similar sized airways for accurate assessment of changes and we speculate this finding will be relevant across other species as well.Support or Funding InformationNAThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of Acute Cadmium Exposure on the Vascular Endothelial Barrier Antigen and Astrocyte Morphology in the Developing Rat Central Nervous System
Authors: Ibiwoye Michael O , Matthews Qiana , McElroy Pamela , Foster James D 
Published: June 2021

Cadmium is a non‐biodegradable heavy metal and a major worldwide environmental pollutant. Clinical and experimental studies have documented the neurotoxic and behavioral effects of cadmium. However, the exact pathophysiologic mechanism(s) of the toxicity of cadmium for the human central nervous system is not completely understood. A literature review indicates that disruption of the blood‐brain barrier (BBB) structures is a key event in the pathogenesis of central nervous system disorders including cadmium neurotoxicity and infectious diseases. Several BBB‐associated proteins, including occludins, junctional adhesion molecules and claudins, among others, have been identified and used for evaluating BBB disruption under pathological conditions. A developmentally regulated rat blood‐brain barrier (BBB)‐specific capillary endothelial marker, the endothelial barrier antigen (EBA), has been previously isolated and classified by Sternberger and others. In previous studies, we have demonstrated that anti‐EBA exclusively recognizes barrier‐competent microvessels in the rat central and peripheral nervous system (CNS and PNS) and is negative in endothelial cells of peripheral tissues such as the heart and kidney. The antigen is also absent in brain microvessels possessing fenestrated endothelia such as the median eminence, organum vasculosum of lamina terminalis (ovalt) and pineal gland. In our current study, we applied a double, sequential indirect immunofluorescent protocol with anti‐EBA and an antibody directed against glial fibrillary acidic protein (GFAP), to examine the immunoreactivity patterns and morphological alterations in BBB microvessels and astrocytes, following acute exposure to a single, high dose of cadmium in young rats. We detected a moderate reduction in immunoreactivity and number of microvessels labelled by the anti‐EBA in the forebrain, cerebellum and midbrain in cadmium‐exposed rats compared to normal controls. Within the white and gray matter of cadmium‐exposed brains, we observed weakly GFAP‐reactive astrocytes displaying cell bodies with ill‐defined borders and blurry cytoplasm. The astrocyte nuclei appeared markedly enlarged, intensely hyperchromatic and exhibited marked chromatin condensation with varying degree of nuclear fragmentation. The weakly GFAP‐positive cell processes were hypertrophic and showed less branching compared to the strongly GFAP‐positive, highly branched astrocytes. Thus, indicating that EBA is involved in the pathogenesis of cadmium neurotoxicity in the rat model system.Support or Funding InformationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐inflammatory activity of Sageretia thea in LPS‐stimulated RAW264.7 cells
Authors: Park Gwang Hun , Eo Hyun Ji , Jeong Jin Boo 
Published: June 2021

Sageretia thea (S. thea) has been reported to exert various pharmacological activities such as anti‐oxidation, anti‐cancer and anti‐human immunodeficiency virus activity. However, there is little study on the anti‐inflammatory activity of S. thea. Thus, we evaluated the anti‐inflammatory effect of extracts of leaves (ST‐L) and branches (ST‐B) from S. thea in LPS‐stimulated RAW264.7 cells. ST‐L and ST‐B significantly inhibited the production of the pro‐inflammatory mediators such as NO, iNOS, COX‐2, IL‐1β and IL‐6 in LPS‐stimulated RAW264.7 cells. ST‐L and ST‐B blocked LPS‐induced degradation of IκB‐α and nuclear accumulation of p65, which resulted to the inhibition of NF‐κB activation in RAW264.7 cells. ST‐L and ST‐B also attenuated the phosphorylation of ERK1/2, p38 and JNK in LPS‐stimulated RAW264.7 cells. In addition, ST‐L and ST‐B increased HO‐1 expression in RAW264.7 cells, and the inhibition of HO‐1 by ZnPP reduced the inhibitory effect of ST‐L and ST‐B against LPS‐induced NO production in RAW264.7 cells. Inhibition of p38 activation and ROS elimination attenuated HO‐1 expression by ST‐L and ST‐B, and ROS elimination inhibited p38 activation induced by ST‐L and ST‐B. ST‐L and ST‐B dramatically induced nuclear accumulation of Nrf2, but this was significantly reversed by the inhibition of p38 activation and ROS elimination. Collectively, our results suggest that ST‐L and ST‐B exerts potential anti‐inflammatory activity by suppressing NF‐κB and MAPK signaling activation, and activating HO‐1 expression through the nuclear accumulation of Nrf2 via ROS‐dependent p38 activation. These findings suggest that ST‐L and ST‐B may have great potential for the development of anti‐inflammatory drug to treat acute and chronic inflammatory disorders.Support or Funding InformationThis work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF‐2016R1D1A3B03931713 and NRF‐2018R1A6A1A03024862), and by a grant from National Institute of Forest Science in 2019.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction of Activator of G‐protein Signaling 3 (AGS3) and the signaling protein Dishevelled: Implications for the regulated distribution AGS3 within the cell and its functional diversity
Authors: VURAL ALI , LANIER STEPHEN 
Published: June 2021

Activator of G‐protein Signaling 3 (AGS3), a receptor independent activator of G‐protein signaling, consists of 7 tetratricopeptide repeats (TPR) upstream from 4 G‐protein regulatory motifs (GPR), each of which interact with GαGDP. AGS3 moves among multiple cellular compartments including cytosol, the cell cortex, the Golgi apparatus, the aggresomes and cellular puncta, with suggested oscillation between hydrophilic and hydrophobic domains. Such a regulated subcellular distribution is intimately related to the functional diversity of the protein, which remains mechanistically elusive. We recently reported that a mutation of a single threonine residue in the AGS3‐GPR domain alters the apparent phosphorylation status of the protein and results in the localization of AGS3‐T602A to punctate subcellular structures in contrast to the more diffuse, cytosolic distribution observed with WT AGS3. Moreover, these puncta are distinct from defined intracellular organelles or vesicles and their localization is regulated by interaction with AGS3 binding partners and potentially by the phosphorylation status of AGS3. In a search for proteins that exhibited a similar punctate distribution, we determined that the Dishevelled protein 2 (Dvl‐2) exhibits a punctate cellular distribution that is actually a “mirror” image of that observed with the AGS3‐T602A. However, co‐expression of AGS3‐T602A with Dvl‐2 indicated that the two proteins were localized to distinct punctate populations in the cell. In contrast AGS3‐WT, which typically has a non‐homogeneous, diffuse distribution in the cytosol, was actually redistributed to Dvl‐2 puncta upon co‐expression of the two proteins. The Dvl‐2 induced localization of WT AGS3 to puncta was reversed by coexpression of Giα3, which likely reflects the interaction of Giα3 with the AGS3 GPR domains. This series of observations suggest that the positioning of AGS3 as part of the Dvl‐2 puncta may be phosphorylation‐dependent and regulated by G‐proteins. In addition, these observations taken together, suggests a potential role for AGS3 in canonical and noncanonical WNT signaling pathways involving Dvl‐2.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aging influences cerebrovascular function via BK channel in a sex‐specific manner
Authors: Pabbidi Mallikarjuna , Pareek Tanya , Reed Joseph , Sontakke Sumit 
Published: June 2021

The myogenic response of middle cerebral arteries (MCA) protects the brain by altering the diameter in response to lumen pressure to maintain constant cerebral blood flow. Large conductance potassium (BK) channels are known to regulate the myogenic response. However, it is not clear how aging will influence the myogenic response via altering BK channel function in males and females. Here, using vascular reactivity, patch‐clamp and biochemical methods, we found that age‐associated changes in BK channel subunit expression and function alter myogenic response in a sex‐specific manner. Aging exaggerated the myogenic response of MCA of females but did not change the myogenic response in males. Aging abolished differences in myogenic tone of MCA (MT as determined from active and passive diameters) in males and females. Aging, while decreased wall to lumen ratio in females, it increased in males revealing a sex‐specific difference in MCA remodeling. Aging while decreased total K channel and BK channel currents in vascular smooth muscle cells (VSMCs) isolated from female vessels, it did not affect males. Spontaneous transient outward currents (STOC) that represent BK function are also decreased with aging in both males and females. However, the magnitude of the decrease in STOC amplitude and frequency is much greater in aged females than aged males. While aging increased BKα subunit, it decreased β1 subunit protein in cerebral vessels of both males and females. Age‐associated increase in BKα subunit is similar between males and females. However, there is a greater and significant reduction of BKβ1 subunit protein in aged females than aged males. Activation of BKα and β1 subunits either individually or both failed to decrease MT of MCA isolated from aged males and females at physiological lumen pressures. Besides, aging diminished acute estradiol (17βE)‐mediated decrease in MT either alone or combined with BKβ1 agonist in females. However, this combination significantly decreased MT in ovariectomy females that are chronically supplemented with 17βE via increasing BKβ1 protein content. Overall our results suggest that age‐associated changes in cerebral vascular structure and function and that of BK channel subunit expression and function are sex‐specific. Chronic supplementation of 17βE increases BKβ1 protein content, and its activation by agonist combined with acute 17βE attenuates MT in aged females.Support or Funding InformationSupported by AHA SDG (13SDG14000006) and IRSP from UMMC to Mallikarjuna R. Pabbidi.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.