bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lactobacillus paracasei PS23 modulated the age‐related inflammation in Senescence Accelerated Mouse Prone 8 (SAMP8) mice
Authors: Huang Hui‐Yu , Huang Ying‐Xu , Chen Yi‐Wen , Chou Nien‐Wei , Chen Yi‐Rong , Lee Hui‐Chun , Lee Yi‐Ju 
Published: June 2021

Sarcopenia is a common impairment in the elderly population responsible for poor outcomes later in life, which is enhanced by inflammation. Only a few strategies have been reported to reduce sarcopenia. We investigated whether Lactobacillus paracasei PS23 (LPPS23) decelerates age‐related muscle loss and its inflammatory response; LPPS23 has been reported to delay some age‐related disorders. Female senescence‐accelerated mouse prone 8 (SAMP8) mice were divided into three groups (n = 6 each): non‐aging (16‐week‐old), control (28‐week‐old), and PS23 (28‐week‐old) groups. The control and PS23 groups were given saline and LPPS23, respectively. We evaluated the effects of LPPS23 by analyzing body weight and composition, reactive oxygen species, and IL‐6. The positive anti‐ROS effect of LPPS23 in the aged SAMP8 mice demonstrated that LPPS23 may protect muscle mitochondria against ROS damage, which may be associated with a lower level of sarcopenia in aged SAMP8 mice. Similar to LPPS23, some probiotics has been reported to have anti‐ROS properties; more probiotics may be also proofed to use for modulating sarcopenia in the future.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acrolein modifies protein functions and induces tissue damage at advanced age
Authors: Uemura Takeshi , Igarashi Kazuei 
Published: June 2021

Acrolein (CH2=CHCHO) is a highly reactive unsaturated aldehyde generated mainly from oxidation of spermine, one of polyamines, in vivo. Acrolein causes strong oxidative stresses at sub microM concentration that is quite lower than reactive oxygen species (ROS). The activity of spermine oxidase, the enzyme involved in the generation of acrolein, is increased with age. Thus, acrolein plays important roles in tissue damage at advanced age such as brain stroke, dementia, renal failure and autoimmune diseases. As global society ages, it is required to prevent aging related diseases to maintain the quality of life (QOL) of the elderly. The objective of the presenting study is to identify the novel target of acrolein toxicity which can be a target for the prevention of age related disorders.We have analyzed proteins modified with acrolein in cells and identified GAPDH (glyceraldehyde‐3‐phosphate dehydrogenase), MMP‐9 (matrix metalloprotease 9), tubulin and GSTπ (glutathioneS‐transferase π). GAPDH was inactivated by acrolein modification that caused a decrease in cellular ATP content and induction of apoptosis through nuclear translocation. On the other hand, MMP‐9 was activated by acrolein to induce tissue damage in Sjogren's syndrome patients. Acrolein modification of tubulin inhibited polymerization of microtubules and disrupted cytoskeletal structures, then cell division and development of neuronal axons. GSTπ catalyzes conjugation of acrolein with glutathione in cells to neutralize it. GSTπ was modified and inactivated by acrolein to form polymer of proteins.In conclusion, we have found that through modification of GAPDH, MMP‐9, tubulin and GSTπ proteins, acrolein caused tissue damage in age related disorders. Acrolein scavengers can prevent the modification of proteins and further tissue damage.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Berberine inhibits free fatty acid and LPS‐induced inflammation via modulating ER stress response in macrophages
Authors: Wang Yanyan , Yan Joanna , Chen Weidong , Zhou Huiping 
Published: June 2021

Inflammation plays a pivotal role in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Berberine (BBR), an isoquinoline alkaloid isolated from many medicinal herbs, has been used to treat various diseases including liver diseases for hundreds of years. Our previous studies have shown that BBR inhibits high fat‐diet‐induced steatosis and inflammation in rat model. However, the underlying molecular mechanisms remain to be identified. This study was to identify the potential mechanism by which BBR inhibits free fatty acid (FFA) and LPS‐induced inflammatory response in mouse macrophages.MethodsMouse RAW2674.1 macrophages were treated with palmitic acid (PA) or LPS or PA+ LPS with or without different concentrations of BBR (0–10 μM) for different periods (0–24h). The mRNA and protein levels of IL‐1β, IL‐6, TNF‐α, MCP‐1, COX‐2, and ER stress genes (CHOP, ATF4, XBP‐1) were detected by real time RT‐PCR, Western blot and ELISA, respectively.ResultsBBR significantly inhibited PA‐ and LPS‐induced activation of ER stress and expression of proinflammatory cytokines (IL‐1β, IL‐6) and COX‐2. PA/LPS‐mediated activation of ERK1/2 was inhibited by BBR in a dose‐dependent manner.ConclusionBBR inhibits PA/LPS‐induced inflammatory responses through modulating ER stress‐mediated ERK1/2 activation in macrophages.Support or Funding InformationThis work was supported by VA Merit Award I01BX004033 and 1I01BX001390, VA Career Scientist Award IK6BX004477 and National Institutes of Health Grant R01 DK104893 and R01DK‐057543This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigation of the Relative Activity of Monomer and Dimer SecYEG Translocons
Authors: Strauss Alexa , Banerjee Tithi , Oliver Donald 
Published: June 2021

The general secretory (Sec) pathway is the principal protein transport pathway in bacteria. Two key components of this system are the protein complex SecYEG, which forms a protein‐conducting channel within the plasma membrane, and the ATPase motor protein SecA, which drives the protein translocation process. Whether the SecYEG complex functions as a monomer or requires dimerization with a neighboring SecYEG complex for activity remains unclear. To address this important question, we have made use of both photobleaching and total internal reflection fluorescence (TIRF) microscopy to assess the fluorescently labeled SecY oligomeric state, as well as an in vitro protein translocation assay to assess Sec‐dependent protein transport activity. The key component of our system is the SecYEG proteoliposome, which is a lipid vesicle reconstituted solely with SecYEG protein. In this study, we reconstituted proteoliposomes at different lipid to SecYEG protein ratios to produce distinct vesicle populations containing either mostly SecYEG monomers or containing about half SecYEG monomers and half SecYEG dimers. Next, we utilized TIRF microscopy to determine the percent of singly or doubly reconstituted proteoliposomes existing within a given population based on the ratio of single or double photobleaching events. Finally, we optimized and performed an in vitro protein transport assay to assess the overall protein translocation activity of a given population. Initial findings from theses assays suggest that the monomer SecYEG proteoliposome population is more efficient at translocating protein compared to the population with substantial dimer content. Further studies will be required to fully confirm this finding. Ultimately, we hope to elucidate any differences in the protein translocation activity between these distinct populations in order to better understand the functionality of monomer and dimer SecYEG translocons in this universally conserved system.Support or Funding InformationFunding generously provided by the NIH through the National Institute of General Medical Sciences (GM110552)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
GLP‐1 Receptor Is Associated with Genes Involved in Fatty Acids Oxidation and White‐to‐Brown Fat Differentiation in Epicardial Adipose Tissue (EAT)
Authors: Romanelli Massimiliano Marco Corsi , Vianello Elena , Malavazos Alexis Elias , Tacchini Lorenza , Schmitz Gerd , Iacobellis Gianluca , Dozio Elena 
Published: June 2021

Epicardial adipose tissue (EAT) is an important risk factor for cardiovascular diseases (CVD). Recent clinical evidence suggested that the potential beneficial effects of glucagon‐like peptide 1 (GLP‐1) analogs on CVD may deal with the reduction of EAT amount, possibly by targeting EAT GLP‐1 receptor (GLP‐1R), as recently discovered. Nevertheless, the role of EAT GLP‐1R and its interplay with EAT genes involved in adipogenesis and fatty acids metabolism are presently unknown.EAT was collected from 17 patients with coronary artery disease (CAD) undergoing elective coronary artery bypass graft for microarray analysis of GLP‐1R and genes involved in fatty acid metabolism and adipogenesis. GLP‐1 plasma levels were measured by enzyme‐linked immunosorbent assay. EAT thickness was measured by echocardiography. The study has been performed in accordance with the Principles of Declaration of Helsinki.EAT GLP‐1R was directly correlated with genes promoting beta‐oxidation and white‐to‐brown adipocyte differentiation, and inversely with pro‐adipogenic genes. Circulating GLP‐1 levels were higher in CAD than healthy subjects and in patients with greater ultrasound‐measured EAT thickness.GLP‐1 and its analogs may target EAT GLP‐1R and therefore reduce local adipogenesis, improve fat utilization and induce brown fat differentiation. The increase in circulating GLP‐1 levels may be a compensatory mechanism to improve metabolic dysfunctions. As EAT lies in direct contiguity to the myocardium and coronary arteries, the beneficial effects of GLP‐1 activation may extent to the heart.Support or Funding InformationThe study was supported by funding from Fondazione E. A. Fiera Internazionale di Milano to Università degli Studi di Milano and Ricerca Corrente funding from Italian Ministry of Health to IRCCS Policlinico San Donato.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CRISPR/Cas9‐Mediated Rescue of Shiverer Mice (MBPShi) via AAV9 Delivery across the Blood‐Brain Barrier
Authors: Ille Alexander , Lamont Hannah , Ruvio Stacy , Bernosky‐Smith Kimberly 
Published: June 2021

The shiverer mouse is a genetically modified strain of M. musculus which exhibits ataxia, tremor, and seizure, among other symptoms, due to impaired CNS function. The strain is generated through deletion of a large portion of the Myelin Basic Protein (MBP) gene. Due to lack of MBP, the shiverer mouse presents with hypomyelination and the corresponding “shivering” phenotype. To date, shiverer mouse rescue has been accomplished through embryological stem‐cell implant, or gene delivery via zygote microinjection. However, these methods are limited to rescue of the subsequent generation and cannot be applied to a mouse of the current generation. Another obstacle to consider when exploring pathologies affecting the CNS is the ability of a therapeutic candidate to cross the blood‐brain barrier (BBB). Here we propose a novel method for the genetic rescue of current‐generation shiverer mice. This method involves CRISPR‐mediated insertion of a functional MBP transgene via the delivery of an AAV9 delivery mechanism. This involves a simple procedure of non‐specific intravenous injection of the mouse, relying on viral delivery across the BBB and bypassing the need for invasive surgical intervention.Support or Funding InformationD'Youville CollegeThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structure‐Activity Relationships of Selective Pyrimidine Agonists on α7‐nAChRs
Authors: Hernandez Gisela Andrea Camacho , Kaczanowska Katarzyna , Brandao Stefania , Stokes Clare , Papke Roger Lee , Taylor Palmer 
Published: June 2021

Nicotinic acetylcholine receptors (nAChRs) are prototypic pentameric ligand‐gated ion channels (pLGIC), activated by the endogenous neurotransmitter acetylcholine. nAChRs are present in the peripheral and central nervous systems, where they modulate central, peripheral autonomic and neuromuscular functions. One of the most abundant neuronal nAChRs is the homopentameric α7. This subtype comprises of five identical α7 subunits with five binding sites, located between the subunit interfaces. The α7‐nAChR has a relatively low affinity for nicotine and acetylcholine, and a high permeability for Ca+2. Activation occurs on sub‐millisecond time scale; desensitization occurs upon prolonged application of agonist. nAChRs are found on presynaptic terminals in brain mediating the release of other neurotransmitters.We previously synthesized a family of 4,6 substituted 2‐aminopyrimidines; our lead candidate, AC‐171A2, activates selectively the α7‐nAChRswith an EC50 of 70nM in the presence of PNU‐120596, diverging from classical agonist on binding interaction as well as chemical components. This compound has an EC50 of 360nM when fast activation was evaluated in a Xenopus oocyte system without PNU‐120596. When recovery of the receptor was assessed, AC‐171A2 allowed full recovery at 3mM concentration after 12 min. One of the goals of our current study is to better understand the molecular determinants of these ligands in these different time frames. We have developed an expanded library of compounds, based on previous leads, by combining the pyrimidine system with the di‐picolyl substitution at the 4‐position. To distinguish receptor interactions on nAChRs and related pentameric ligand‐gated ion channels, we employ HEK cells transfected with cDNA's encoding one of three requisite receptor subtypes: α7‐nAChR, α4β2‐nAChR and 5HT3AR, along with a fluorescent reporter, TN‐XXL. To assess fast activation of lead candidates, we use a Xenopus oocyte system expressing α7‐nAChR. This approach has enabled us to define the molecular determinants of the selective activation of α7‐nAChR's and offer more selective and potent lead candidates with EC50's extending to 30nM in the presence of PNU‐120596.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 TMEM16A Cl− Channel by Fungus‐Derived Penicillide in Airway Epithelial Cells
Authors: Yimnual Chantapol , Satitsri Saravut , Rukachaisirikul Vatcharin , Muanprasat Chatchai 
Published: June 2021

Asthma represents one of the most common chronic diseases affecting around 300 million people with 250,000 annual deaths worldwide. It is characterized by airway inflammation, airway constriction, and mucus overproduction. The current treatments of asthma focus only on the first 2 characteristics without targeting on mucus overproduction. Transmembrane protein 16A (TMEM16A) is a Ca2+‐activated Cl− channel that, when activated, causes mucus production and secretion. Inhibition of TMEM16A activity effectively reduces mucus production along with its secretion. However, available TMEM16A inhibitors so far are synthetic compounds, or derived from plants which have limitation of large‐scale production. Therefore, this study aimed to identify novel TMEM16A inhibitor from fungal resources. A screening of >400 fungus‐derived natural compounds showed that N22, derivative of penicillide, potently inhibited ATP‐induced TMEM16A Cl− current with IC50 ~ 2 μM as analyzed by short‐circuit current measurement in human airway epithelial (Calu‐3) cell monolayers. Moreover, this inhibition was observed when TMEM16A is activated by Ionomycin (Ca2+ ionophore), or Eact (specific TMEM16A activator), suggesting that this inhibition was downstream to elevation of intracellular Ca2+ and might directly on TMEM16A molecule. As analyzed by MTT assays, inhibitory effect of N22 on TMEM16A‐mediated Cl− transport was not associated with cytotoxicity in Calu‐3 cells. Collectively, penicillides represent the first class of TMEM16A inhibitor from the fungi which may be beneficial for treatment of asthma resulting from mucus overproduction and secretion.Support or Funding InformationThis work was supported by the NSTDA Chair Professor grant (the Fourth Grant) of the Crown Property Bureau Foundation, Faculty of Science Mahidol University, Faculty of Graduate Studies Mahidol University and Medical Scholars Program, Mahidol 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.
Developing Inhibitors that Exploit ABCG2 and Cancer Dependencies to Improve Therapeutic Outcome
Authors: Fukuda Yu , Allcock Alice , Wijaya Juwina , Gose Tomoka , Lin Wenwei , Chen Taosheng , Schuetz John D. 
Published: June 2021

Multidrug efflux transporters such as ABCG2 limit drug penetration into cancer cells and limit therapeutic effectiveness of many chemotherapeutic drugs. We have shown previously that inhibition of ABCG2, when combined with a conventional chemotherapy, improved treatment efficacy in murine models of acute myeloid leukemia and medulloblastoma (1,2). In this study, we sought to identify ABCG2 inhibitors that are also capable of targeting a cancer cell‐specific pathway. By combining these ABCG2 inhibitors with conventional chemotherapeutic drugs that are ABCG2 substrates, we anticipate synergistic cytotoxic effects that will produce superior treatment outcomes. A high‐throughput functional ABCG2 inhibition screen was performed using a library containing ~12,000 bioactive compounds including FDA‐approved drugs. Hit compounds were validated in a dose dependent manner, and compounds that were cytotoxic and had a low IC50 (< 1 μM) for ABCG2 inhibition were selected for further investigation. We identified kinase inhibitors (KIs) that have not been previously reported to be ABCG2 inhibitors and confirmed the direct interaction of one KI to ABCG2 using a membrane protein cellular thermal shift assay (CETSA). We investigated if the combination of a conventional cancer chemotherapeutic drug, mitoxantrone, and the KI produced an increase in efficacy against primary leukemic cells. In vitro testing of this combination on murine myeloid leukemia cells produced synergistic cell killing with an IC50 value ~10‐fold lower for each compound compared to the IC50 value for normal hematopoietic progenitors. Murine leukemia cells were then treated either singly or with the mitoxantrone‐KI combination ex vivo, followed by transplantation into lethally irradiated congenic recipient mice. The mice that received cells treated with mitoxantrone and the KI had a significant increase in their overall survival. We are extending our findings to other cancers that overexpress ABCG2 as this may be a viable therapeutic approach that improves treatment response in cancers that specifically express ABCG2 and are treated with ABCG2 substrates.Support or Funding InformationThis work was supported by NIH and by the ALSAC.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Contribution of the Renal Lymphatic Circulation to the Development of Salt‐Sensitive Hypertension
Authors: Garner Brittney R , Stolarz Amanda J , Rusch Nancy J , Mu Shengyu 
Published: June 2021

More than one billion people worldwide suffer from hypertension, which is a leading cause of stroke and cardiovascular disease. According to the National Heart, Lung, and Blood Institute (NHLBI) Working Group, an estimated 40% to 85% of hypertensive patients exhibit salt‐sensitivity, which increases the risk for cardiovascular events and mortality. Salt‐sensitive hypertension is known to be caused by an imbalance between sodium retention and excretion in the kidney. However, the mechanisms that enable the kidney to deliver excessive re‐absorbed salt and volume back to the systemic circulation remain obscure. Interestingly, recent studies indicate that the peripheral lymphatics act as a “salt sink” to store excessive salt and volume, thereby maintaining sodium homeostasis to regulate blood pressure. This finding led us to consider whether renal lymphatics contribute to excessive sodium retention in the kidney during the development of salt‐sensitive hypertension. To test this hypothesis, we initially evaluated lymph‐angiogenesis in the kidneys of rats with or without salt‐sensitive hypertension. Real‐time PCR, immunohistochemistry and Western blots revealed increased expression of the lymphatic‐specific marker VEGFR3 in the kidneys of Dahl salt‐sensitive (DSS) rats fed a high‐salt diet compared to either DSS rats fed regular chow or Dahl salt‐resistant (DSR) rats with or without high‐salt intake. To explore the impact of renal lymph‐angiogenesis on blood pressure, we surgically ligated the renal collecting lymph vessel (LV) in DSS rats. Ligation of the renal LV in DSS rats attenuated the increase in systolic blood pressure caused by a high‐salt diet. Notably, a growing body of evidence has implicated macrophages in lymph‐angiogenesis. Indeed, we also observed that a high‐salt diet promoted renal infiltration of macrophages, which stimulated renal lymph‐angiogenesis via upregulating VEGFR3 and its ligand VEGFC. Collectively, our data suggest that in response to a high‐salt diet, kidney‐infiltrated macrophages stimulate renal lymph‐angiogenesis, which appears to support the development of salt‐sensitive hypertension.Support or Funding InformationSupported by NIH grants T32 GM106999 (BRG) and R21 CA187325‐01A1 (NJR), and a Medical Research Endowment Award (SM) from the Univ. of Arkansas for Medical Sciences.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.