bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Photobiomodulation Effects in Cardiac Hypertrophy in Experimental Model of Myocardial Infarction
Authors: Costa Leandro Oliveira D'Alcantara , Atum Allan , Serra Andrey , Manchini Martha , Feliciano Regiane S. , Silva José Antônio A. 
Published: June 2021

IntroductionThe process of cardiac remodeling is characterized by changes in geometry, volume, mass and constitution of the heart in response to tissue aggression. Myocardial hypertrophy is an important component of cardiac remodeling, which allows the heart to maintain its basic functions under increasing load conditions. The renin‐angiotensin system plays an important role in the pathophysiology of myocardial infarction and cardiac hypertrophy and contributes to the progression of heart failure.ObjectiveTo evaluate the effect of photobiomodulation in genes participating in the cardiac hypertrophy process, by the analysis of Edn1, Ace, Agtr1a and Ace2 mRNA.Materials and MethodsFifteen female Wistar rats were divided into three groups: Sham (5), IM (5) and IM + Laser (5) (Ethics Committee: 0015–2012). The infarction induction was made by occlusion of the descending left coronary artery, the laser being applied for 60 seconds at the occlusion site immediately after the infarction. Left ventricle samples from the experimental animals were collected 72h after infarction. Modifications of the gene expression of the genes: Edn1, Ace, Agtr1a and Ace2 were analyzed by Real‐Time PCR.ResultsThe IM group determined a significant increase in expression of the genes: Edn1, Ace and Agtr1a when compared to the Sham group 3 days after the infarction. Laser therapy significantly reduced the expression of these genes when compared to the IM group: Edn1, Ace and Agtr1a when compared to the IM group, suggesting a reduction in hypertrophic processes after laser therapy.ConclusionIncreased ACE expression in experimental models of myocardial infarction, as well as increased endothelin, are related to hypertrophic processes after myocardial infarction. Our studies suggest the action of the laser in the infarcted myocardium through the participation of the renin angiotensin system with the reduction of the expression of the genes involved in these processes.Support or Funding InformationFAPESP, CNPq, CAPES, UNINOVEThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of Gαi protein selective μ opioid receptor analgesics to limit side‐effects
Authors: Liu Jianxi , Kenny Paul J. 
Published: June 2021

The clinical utility of opioids as analgesics is limited by their serious side effects, which include respiratory inhibition, gastrointestinal motility disorders, tolerance and addiction. Opioids act by binding to μ opioid receptors (MORs) to trigger G‐protein and β‐arrestin‐mediated signaling cascades. The Gαi class of G‐protein is thought to play an important role in transducing the analgesic actions of opioids. Gαi subunits include Gi1, Gi2, Gi3, Go and Gz. In addition, Gβγ subunits also contribute to the actions of opioids, and the many subtypes of Gβγ subunits adding yet more complexity to MOR‐mediated signaling.Gαi proteins show discrete tissue and brain distributions, each subunit has discrete enzymatic kinetics, and these subunits are differentially regulated by GTPase‐activating proteins (GAPs), Regulator of G protein (RGS) proteins, and other effectors. Moreover, findings from genetically modified mice suggest that disruption of specific Gαi proteins results in discrete patterns of altered responses to opioids. Based on these observations, we hypothesize that compounds can be developed to modulate specific Gi protein coupling to μ receptors, which will result in analgesic actions with less pronounced side‐effects.Currently, novel opioids are developed by structure‐based or Gqi5‐mediated calcium readout. Consequently, Gqi5 biased opioids are likely to be preferred. To identify functionally novel opioids, we constructed MOR/Gqi15, i35, o5 and z5 in HEK 293 cells. We profiled ten reference compounds: DAMGO, β‐endorphin, met‐enkephalin, morphine, hydrocodone, oxycodone, TRV130, PZM21, SR17018 and fentanyl in these cell lines. We found that the majority of opioids signal through all Gi proteins with similar potency and efficacy. However, a minority showed strong Gi protein bias. Finally, because neuronal excitability is mediated by Gβγ subunits acting at N‐type CaV and G protein gated inwardly‐rectifying potassium (GIRK) channels, we generated MOR/GIRK1/2 and MOR/CaV2.2 α1/α2δ1/β1 in HEK 293 cells. These lines will be used to identify molecules that influence Gβγ actions on the activity of these effector channels.These preliminary data demonstrate that opioids are likely signal in manner that can be biased to specific Gi proteins. This offers an opportunity to differentiate/discover compounds with unique pharmacodynamic profiles. Novel opioids that act through discrete Gi protein, or on Gβγ proteins, will facilitate greater understanding about the relationship between selective G‐protein coupling and opioid‐related side‐effects. We aim to leverage this new information to develop novel opioid analgesics with limited side‐effects.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RB and CDKN2A cause Rapamycin resistance in cancer cells
Authors: Chakraborty Sohag , Utter Matthew , Foster David 
Published: June 2021

mTOR‐the mammalian target of Rapamycin acts as the central regulator of multiple cellular processes like cell growth, proliferation and survival by integrating signals via nutrients, growth factors, hormones and energy sensing. In cancer cells, the mTOR pathway is highly dysregulated providing survival signals to the cells for their uncontrolled growth. Hence, mTOR has been a potential therapeutic target for cancer treatment for the past two decades. Application of high micro‐molar doses of Rapamycin has been found to successfully inhibit mTORC1 by blocking the phosphorylation of both downstream substrates ‐ S6K and 4E‐BP1. Our lab previously reported that Rapamycin acts as a cytostatic drug and causes G1 cell cycle arrest in the presence of TGF‐β signal. However, in the absence of TGF‐β signal, the drug induces apoptosis. The rationale behind the apoptotic effect of Rapamycin in absence of TGF‐β is that without TGF‐β signals, cells do not arrest in G1 and progress into S phase where Rapamycin is cytotoxic rather than cytostatic. Of significance, we found that cancer cells with nonfunctional RB and mutated CDKN2A do not undergo apoptosis upon Rapamycin treatment in the absence of TGF‐β signal. Proteins encoded by RB (pRb) and CDKN2A (p16INK4A and p14ARF) are upstream effectors of E2F family of transcription factors that are critical for G1 to S phase progression.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aplysinopsins Inhibit Growth of Crypotococcus neoformans by Interfering with Inositol Metabolism
Authors: Moss Jackson , Clouse Alexandria , McCullough Donna , Wloch‐Salamon Dominika , Johnson Paul , Bialonska Dobrusia 
Published: June 2021

Cryptococcus neoformans is a widespread microscopic fungus (yeast), that humans are frequently exposed to via inhalation. Such exposure is usually harmless, resulting in no adverse reactions. However, for those with a weakened immune system, particularly those with advanced HIV/AIDS, C. neoformans may present as an opportunistic, facultative, intracellular, pathogen causing the disease cryptococcosis, defined by meningitis and/or encephalitis. While the advent of antiretroviral drugs has reduced the risk of cryptococcal meningitis for patients in the US and other developed countries, ~250,000 cases of cryptococcal meningitis are reported each year with a resulting ~200,000 deaths, it remains an acute concern for immunocompromised patients in developing nations with limited access to modern healthcare.In this work, we screened three molecules of the tryptophan derived aplysinopsin family of compounds (aplysinopsin, 5‐bromoaplysinopsin, and 6‐bromoaplysinopsin) for their ability to inhibit the growth of the pathogenic yeast C. neoformans. These compounds were originally isolated from marine invertebrates, but have since been synthesized in the lab for this work. All of the studied compounds exhibited antifungal activity with a potency similar to the current front line treatment for cryptococcosis, Amphotercin B, as the identified IC50 for these compounds was between 9–20 μg/mL. In order to further characterize these compounds activity, we also investigated the underlying mechanisms involved in their antifungal activity via genetic approaches.In order to characterize the underlying mechanisms of this antifungal activity, we used Saccharomyces cerevisiae as a model organism. We identified two mutants lacking genes for metabolism of inositol that showed increased susceptibility to these compounds. These mutants were also more susceptible to aplysinopsins, compared to the wild type, indicating that our compounds may be targeting synthesis of inositol, an important precursor for several secondary messengers in fungi. While the mechanisms of Cryptococcus neurotropism remain poorly understood, recent studies have revealed that C. neoformans has evolved sophisticated inositol acquisition systems and inositol is abundant in the human central nervous system. Thus, inositol utilization in C. neoformans, and its likely contribution to Cryptococcus virulence, may represent one example of a common trait for the emergence of pathogens from environmental reservoirs as well as a target for novel antifungal compounds.Support or Funding InformationUNG FUSE grantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
How Entamoeba histolytica goes vampire: the hunt for new genes in the conserved process of trogocytosis
Authors: Feeney Shea E. , Ralston Katherine S. 
Published: June 2021

The eukaryotic pathogen Entamoeba histolytica (histo‐: tissue; lytic‐: dissolving) causes amoebiasis in humans. The profound tissue damage during infection is likely driven by E. histolytica's ability to kill human cells through trogocytosis (trogo‐: nibble). During trogocytosis, amoebae pinch off and internalize bites of membrane and intracellular contents from human cells, which eventually results in cell death. In contrast, amoebae ingest pre‐killed human cells whole via phagocytosis. Beyond E. histolytica, trogocytosis is part of an underappreciated, emerging theme in eukaryotic biology. While there are some shared features between trogocytosis and phagocytosis, we hypothesize that there are differences. Since E. histolytica is capable of both trogocytosis and phagocytosis, it represents an excellent model to dissect the trogocytosis‐specific mechanism. Both phagocytosis and trogocytosis are under feed‐forward gene regulation in E. histolytica. To identify genes differentially involved in trogocytosis versus phagocytosis, E. histolytica were fed live or pre‐killed human cells, respectively, to induce upregulation of genes involved in these processes. RNA‐seq analysis was used to identify genes that changed substantially in comparison to amoebae that had not been fed human cells. Over 1000 genes were found to be significantly differentially expressed between trogocytosis and phagocytosis and, were involved in various processes including cytoskeletal rearrangement. To begin to explore the roles of select genes that were significantly upregulated in ingestion, we are currently generating RNAi knockdown mutants and evaluating their phenotypes. This work should have broad impact since trogocytosis is directly relevant to amoebiasis and also appears to be a fundamental form of eukaryotic cell‐cell interaction.Support or Funding InformationNIH MCB T32 GM007377 (to SEF); Pew Charitable Trusts (to KSR)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early Alterations in the Expressions of MCTS and HIF‐1α in Experimental Diabetes Mellitus
Authors: Encinas Jéssica Freitas Araujo , Perez Matheus Moreira , Peiró Carlos Henrique Foncesca , Petri Giuliana , Simões Diogo Pimenta , Mathia Vanessa Lopes , Mendonça Maria Isabel Sacchi , da Costa Aguiar Alves Beatriz , Fonseca Fernando Luiz Affonso , da Veiga Glaucia Luciano 
Published: June 2021

AimTo evaluate alterations due to the establishment of experimental diabetes mellitus (DM) alloxan‐induced in biochemical parameters and the expression of genes MCT1, MCT4, CD147 and HIF‐1α in different tissues and in the blood profile.MethodsWistar rats (120–180g) were conditioned at the FMABC bioterium in conditions of 12 hours light/dark, climate temperature (21 ± 2 °C) and ad libitum supply of water and food. Animals of the DM group were treated with Alloxan monohydrate (120 mg/kg) intraperitoneal (ip); the control group sham (NDS) received injections of sodium chloride 0,9% (ip). The glycemic values were measured weekly during the period of treatment through a commercial glycosimeter by puncturing of the caudal vein of the animal; animals with glycemia above ≥200 mg / dL were determined as diabetic. Evaluated the biochemical parameters: glycemia, blood creatinine and urea, and the expression of molecular biomarkers in the heart, brain, kidney and blood by the RT‐qPCR technique. Data was analyzed by GraphPad Prism® 6.0, statistical differences were observed by the Mann‐Whitney test and Student T test with an established significance level in 5% (descriptive value of p <0.05).ResultsThe biochemical parameters found were: glycemia (DM7 619.1 ± 140.7 * p <0.005, n = 10 vs NDS 192.5 ± 43.64, n = 4), blood creatinine (DM7 0.46 ± 0.05 * p <0.005, n = 10 vs NDS 0.40 ± 0.07, n = 4) and urea (DM7 96.7 ± 26.6 * p <0.005, n = 10 vs. NDS 46.5 ± 7.0, n = 4) these values follow the established standards for the diabetic profile in the present study. In the molecular analyzes an increase of CD147 in the brain was observed (DM7 26.72 ± 27.45, * p <0.005, n = 9 vs NDS 1,489 ± 1,648, n = 4). In the blood profile an enhancement in the expressions of all the genes studied in the animals with DM were found; MCT1 (DM7 3,739 ± 3,057 * p <0.005, n = 10 vs NDS 0.009745 ± 0.007489, n = 4); MCT4 (DM7 7.364 ± 7.3, * p <0.005 n = 10 vs NDS 0.06399 ± 0.04844, n = 3); CD147 (DM7 4,295 ± 4,424 * p <0.005, n = 10 vs NDS 0.2212 ± 0.01653, n = 4) and HIF‐1α (DM7 1,413 ± 1,087 * p <0.005, n = 10 vs NDS 0.003833 ± 0.003741, n = 4). A positive correlation between glucose and MCT1 in the blood profile (r = 0.8061, * p <0.05, n = 10) was also noted, Spearman test.ConclusionsPreliminary data demonstrate that even early DM promotes serious biochemical changes accompanied by modifications in the expression of genes involved with glycolytic metabolism and in the cellular hypoxia mechanism. Apparently, the brain is the first tissue affected by these changes.Support or Funding InformationCAPES and CNPqThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Generating highly potent and efficacious antibodies to the cholecystokinin B (CCK‐B) receptor by ribosome display for the treatment of neuropathic pain
Authors: Kunamneni Adinarayana , Montera Marena , Gott Katherine , Durvasula Ravi , Westlund Karin 
Published: June 2021

Development of effective non‐opioid therapeutics tailored to specific chronic pain syndromes is sorely needed. Our recent microchip gene expression profile identified upregulated cholecystokinin B (CCK‐B) receptor in trigeminal ganglia (TG) as one suitable target among many others. Thus, the CCK‐B receptor is an ideal candidate to impact both nociceptive and limbic components of chronic pain. More importantly, its inhibition can reverse opiate tolerance. Based on the literature and the gene expression changes over time, the CCK‐B receptor is implicated as one of the persisting functional targets suitable for development of therapeutics to inhibit chronic pain‐ and anxiety‐related behaviors. Here we describe the isolation and characterization of a panel of CCK‐B receptor specific antibodies using ribosome display technology. Furthermore, we demonstrate in vivo efficacy of our antibodies in a relevant model of chronic neuropathic pain. This is the first example, to date, of scFv antibodies that can block the CCK‐B receptor directly with high potency and specificity. This highlights the exciting potential therapeutic opportunity of an antibody for blocking this type of receptor.Support or Funding InformationThis work was supported by Startup dollars from Loyola University Medical Center (RD) and University of New Mexico (KW).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Changes in VEGF and HIF 1α gene expression levels as potential diagnostic/prognostic markers in liquid biopsies on breast cancer patients
Authors: Peiró Carlos Henrique Foncesca , Encinas Jéssica Freitas Araújo , Aquino Glauco Sérgio Avelino , Veiga Glaucia Luciano , Perez Matheus Moreira , Fonseca Fernando Luiz Affonso , Alves Beatriz Costa Aguiar 
Published: June 2021

AimThis study aimed to evaluate the potential diagnostic and/or prognostic value of VEGF and HIF 1α gene expressions in peripheral blood of patients with breast cancer under chemotherapy treatment.MethodsPeripheral blood samples and tumor biopsies were collected from 125 patients with breast cancer and from 25 healthy women (control). Blood samples were collected at diagnosis and after 3 and 6 months of the beginning of chemotherapy. Total RNA isolation was performed using TRIZol reagent. RNA was then converted to cDNA and the gene expression analysis was made by RT‐qPCR. Results were expressed as mean ± standard deviation and analysed by GraphPad Prism® 6.0 using Mann‐Withney, Friedman and Spearman tests, with a significancy level established in 5% (value of p< 0,05).ResultsGene expression analysis showed an increase of HIF 1α gene expression in patients at diagnosis when compared to that of healthy women (Patients: 0.3362 ± 0.7729, n=34 vs Donors: 0.00212 ± 0.001509, n=25, *p<0.0001). Analysis showed that in patients HIF 1α and VEGF expression is higher in peripheral blood than tumor samples (blood: 0.1927 ± 0.4754, n=108 vs Tumor: 0.09498 ± 0.2141, n=91, *p<0.0001 for HIF 1α and blood: 0.001436 ± 0.002469, n=117, Tumor: 0.002364 ± 0.007736, n=91 and *p=0.0041 for VEGF). HIF 1α also showed a differential expression along treatment (at diagnosis: 0.1432 ± 0.2161, n=102; 3 months after chemotherapy: 0.2010 ± 0.3456, n=102 and 6 months after chemotherapy: 0.1657 ± 0.2150, n=102, *p=0.0021). This differential expression along treatment was also observed for VEGF expression (at diagnosis: 0.001314 ± 0.001780, n=112; 3 months after chemotherapy: 0.001891 ± 0.002446, n=112 and 6 months after chemotherapy: 0.001674 ± 0.002260, n=112, *p<0.0001). It was found a correlation between the gene expression levels of HIF 1α and VEGF in peripheral blood (r=05881, *p<0.0001) as well as in the tumor samples (r=0.2804, *p=0.0123).ConclusionsData suggest that HIF 1α could be a diagnosis marker since there's a significant increase in its expression in blood samples from breast cancer patients when compared to healthy women. It's also observed that there's a positive correlation between VEGF and HIF 1α gene expression in peripheral blood samples.Support or Funding InformationFinancing: CNPq (process number: 133505/2018‐9) and FAPESP (Process number: 2017/03558‐3). Associated author: Carlos Henrique Foncesca Peiró Association: ASBMB (Number: 64413).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ate1 Controls Cellular Warburg Effects by Modifying Hif1a with Arginylation
Authors: Moorthy Balaji T. , Jiang Chuanhua , Patel Devang M. , Kumar Akhilesh , Birnbaum Micheal D. , Alfonso Belkis , Huang Jingyu , Barrientos Antoni , Lampidis Theordore J. , Fontanesi Flavia , Zhang Fangliang 
Published: June 2021

The hypoxia‐inducible factor 1a (HIF1a) is a key regulator of stress response and metabolism. Its expression level is known to be controlled by oxygen sensing with two critical proline residues followed by ubiquitination via the pVHL pathway. Here, we report that the degradation of HIF1a is also controlled by N‐terminal arginylation mediated by Arginyltransferase 1 (Ate1) according to the N‐end rule pathway. We found that this mechanism is separate from the classical pVHL‐pathway and it affects cellular Warburg effect, which is the preferred usage of glycolysis that are commonly seen in cancer cells. Furthermore, we found that Ate1, the only arginylation enzyme in animals, appears to have originated from mitochondrial gene transfer. At least a subset of Ate1 is located inside mitochondria and appears to be essential for the composition and functions of respiration complex II. Our study therefore suggest Ate1 as a master regulator of metabolism in cancer cells through arginylation modification of HIF1a.Support or Funding InformationThis study is supported by NIGMS R01 GM 107333, and a start‐up funding from the University of Miami.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.