bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of a Co‐Micronized Composite Containing Palmitoylethanolamide and Polydatin in an Experimental Model of Benign Prostatic Hyperplasia
Authors: Cordaro Marika , Impellizzeri Daniela , Siracusa Rosalba , Gugliandolo Enrico , Fusco Roberta , Esposito Emanuela , Di Paola Rosanna , Cuzzocrea Salvatore 
Published: November 2023

Palmitoylethanolamide (PEA), a fatty acid amide‐signalling molecule has well‐known anti‐inflammatory and neuroprotective effects. Nevertheless, PEA does not possess the ability to prevent free radical formation. Polydatin (PLD), a biological precursor of resveratrol, has antioxidant activity. A combination of PEA and PLD could, conceivably, have beneficial effects on oxidative stress produced by inflammatory processes. In the present study we investigated the effects of a co‐micronized composite containing PEA and PLD (m(PEA/PLD)) in a model of testosterone‐induced benign hyperplasia (BPH).BPH was provoked in rats by daily administration of testosterone propionate (3 mg/kg) for 14 days. This protocol led to alterations in prostate morphology and increased levels of prostaglandin E2 and dihydrotestosterone as well as of 5α‐reductase 1 and 5α‐reductase 2 expression. Moreover, testosterone induced marked inflammation in terms of an increase in nuclear translocation of nuclear factor‐κB p65 and consequently in IκB‐α degradation as well as disregulation of inducible nitric oxide synthase, cyclooxygenase‐2 expression and manganese superoxide dismutase expression and in the apoptosis pathway. Our results show, for the first time, that m(PEA/PLD) is capable of decreasing prostate weight and dihydrotestosterone production in BPH‐induced rats. These effects were most likely correlated to the anti‐inflammatory and apoptotic effects of m(PEA/PLD).Accordingly, these results support the view that m(PEA/PLD) should be further studied as a potent candidate for the management of BPH.
Changes in DNA Damage Response Proteins During Tamoxifen Treatment and Resistance
Authors: Vergne Ralphdy , Ortiz‐Sanchez Carmen , Godoy‐Munoz Lenin Jose , Encarnación Jarline , Matta Jaime 
Published: November 2023

Tamoxifen (TMX) is currently used for the treatment of both early and advanced estrogen receptor positive (ER+) breast cancer in pre‐ and post‐menopausal women. TMX works by blocking the actions of estrogen and preventing proliferation and furthermore, cell death. DNA damage is an alteration of chemical structure of the DNA that activates a network of proteins to start several cellular processes that sense, detect signal and repair DNA lesions. The mechanism of action of several chemotherapeutic drugs is based on damaging the DNA of the cancerous cell. Although TMX's mechanism of action is not through DNA damage, TMX treatment can lead to the production of oxidative stress that in consequence can affect and damage DNA. Our aim is to study the DNA damage induced by TMX resistance in ER+ breast cancer cell line (MCF‐7) cells and assess the expression changes of key proteins involved in this response. Our hypothesis is that there will be more DNA damage in cells treated with TMX in comparison to untreated cells, and the DNA damage contributes to resistance against TMX. To achieve this, we will assess ER expression as an indicator of resistance to TMX, analyze DNA damage during TMX treatment, and study protein expression related to DNA damage using MCF‐7 under TMX. To achieve this, we cultured MCF‐7 cells using complete media without phenol red and 0.5 μM of TMX with 10 μL of insulin. After 1 week, cells were lysed for protein and RNA extraction. Our results shows that ERα expression was similar to untreated cells; therefore, no statistically differences p>0.05 were found. To assess protein expression changes in proteins involved in DNA damage, we assessed PTEN expression by Western blot. Although, no statistically significant differences were found when comparing TMX treated cells with untreated cells (p>0.05) we observed a slight expression change. This preliminary data indicates that PTEN expression might be affected in ER+ breast cancer cells when TMX resistance is developed. Currently, we are developing an MCF‐7 cell line resistant to TMX for additional studies and assessing protein expression changes of other candidates involved in DNA damage response (such as BRCA‐2 and XPF). These results will shed light on the effect of TMX resistance in the DNA damage response of ER+ breast cancer cells. Also, they will allow us to get an inside view and to track the changes in MCF‐7 protein expression during the TMX resistance timeframe.Support or Funding InformationFunded by NIH‐NIGMS #2R25GM096955 (UPRP‐Rise program)Supported by grants from the NCI Center to Reduce Health Disparities and NIH‐NIGMS MBRS Program grant #9SC1CA182846‐04 (J. Matta Laboratory)We also want to acknowledge the support of the Molecular and Genomics Core Laboratory at Ponce Health Sciences University‐Ponce Research Institute (Grant MD00757).
The choline acetyltransferase inhibitor BW813U suppresses the growth of lung adenocarcinoma from smokers
Authors: Akers Austin Tyler , Colclough Kate W , Friedman Jamie R , Bow Eric W , Rimoldi John M , Cutler Stephen J. , Hardmin W. Elaine 
Published: November 2023

The clinicopathological profile of lung adenocarcinoma (LAC) in smokers is completely different from those in non‐smokers. One of the unique features of LAC in smokers is that it is relatively resistant to therapies. For example, targeted therapeutic agents like EGFR‐inhibitors (erlotinib and gefitinib) are highly effective in LACs which develop in non‐smokers. However, these EGFR‐inhibitors display much lower anti‐tumor activity in LACs in patients who are active smokers. Similarly, lung cancer patients (who are active smokers) show lower response to chemotherapy than those who are non‐smokers. However, majority of LAC patients are smokers. This underlines the need to identify novel molecular targets relevant for LAC therapy in patients who are exposed to cigarette smoke via active smoking or exposure to secondhand smoke. Several convergent studies show that nicotine (the addictive component of cigarette smoke) accelerates the growth of lung cancers, as well as confers resistance to chemotherapy. One of the mechanisms underlying these biological activity of nicotine is that it promotes the secretion of the neurotransmitter acetylcholine (ACh) from LAC cells. ACh is known to be an autocrine growth factor for LAC cells and is synthesized by the enzyme choline acetyltransferase (ChAT). The present study investigates the feasibility of ChAT as a molecular target for LAC in smokers. We find that ChAT levels are upregulated in human LAC cell lines and tissues in a manner dependent on the smoking history. Finally, the ChAT inhibitor BW813U causes robust apoptosis in human LAC cells. The magnitude of BW813U‐induced apoptosis is similar across LAC cell lines (irrespective of smoking history); however, the concentration of BW813U which causes apoptosis is lower in LAC cell lines belonging to heavy smokers. Our studies validate choline acetyltransferase (CHAT) as a viable drug target for the majority of population of lung cancer patients who are smokers.Support or Funding InformationFunding for our study was supported by the an NIH R15‐AREA Grant (2R15CA161491‐02), WVU‐MU Health Partnership Grant and by ACTSI Grant from Marshall University. Furthermore, this study was supported in part by an Institutional Development Award (IDeA) Grant number P20GM104932 from the National Institute of General Medical Sciences (NIGMS) and the Research Core B of COBRE, a component of the National Institutes of Health (NIH).
Molecular mechanism of the functions and allosteric regulations of the αβ and αγ heterodimers of human NAD‐dependent isocitrate dehydrogenase
Authors: Ding Jianping , Ma Tengfei , Liu Yabing , Peng Yijie , Huang Wei 
Published: November 2023

Human NAD‐dependent isocitrate dehydrogenase, existing as the α2βγ heterotetramer, catalyzes the decarboxylation of isocitrate into α‐ketoglutarate in the Krebs cycle, which can be allosterically regulated by citrate, ADP and ATP. To explore the functional roles of the individual subunits and heterodimers, we systematically characterized the enzymatic properties of the α2βγ heterotetramer and αβ and αγ heterodimers of NAD‐IDH and performed the structural and functional studies of the αβ and αγ heterodimers. The biochemical and mutagenesis data show that αβ and αγ alone have considerable basal activity but the full activity of α2βγ requires the assembly and cooperative function of both heterodimers. α2βγ and αγ can be activated by citrate or/and ADP, whereas αβ cannot. The binding of citrate or/and ADP decreases the S0.5,ICT and thus enhances the catalytic efficiencies of the enzymes, and the two activators can act independently or synergistically. Furthermore, the allosteric activation of α2βγ is through the γ subunit not the β subunit. These results indicate that the γ subunit plays the regulatory role to activate the holoenzyme, and the β subunit the structural role to facilitate the assembly of the holoenzyme.Structural studies of the αγ heterodimer show that CIT, ADP and Mg2+ bind adjacent to each other at the allosteric site in the γ subunit. The CIT binding induces conformational changes at the allosteric site, which are transmitted to the active site through the heterodimer interface, leading to stabilization of the ICT binding at the active site and thus activation of the enzyme. The ADP binding induces no further conformational changes but enhances the CIT binding through Mg2+‐mediated interactions, yielding a synergistic activation effect. Structural studies of the αβ heterodimer show that the β subunit contains a pseudo allosteric site different from the γ subunit, explaining its incapability to bind activators. However, the αβ heterodimer exhibits similar heterodimer and heterotetramer interfaces as the αγ heterodimer. Mutagenesis and kinetic data show that key residues at the allosteric site and the interfaces play important roles in the activation of the holoenzyme but these at the pseudo allosteric site do not. The structural and biochemical data together indicate that there is a structural communication between the allosteric site of the γ subunit and the active sites of both heterodimers via the interfaces, and the binding of activators to the allosteric site can regulate both α subunits. These findings reveal the molecular mechanism of the functional roles and the allosteric regulations of the αγ and αβ heterodimers of human NAD‐IDH.Support or Funding InformationThis work was supported by grants from the National Natural Science Foundation of China (31170690 and 31521061), the Ministry of Science and Technology of China (2013CB910404) and the Chinese Academy of Sciences (XDB08010302).
A Multifunctional Platform for Rapid Detection and Post‐Detection Analysis of Circulating Tumor Cells
Authors: Yang Yu‐Ping , Datar Ram , Cote Richard , Daunert Sylvia 
Published: November 2023

Circulating tumor cells (CTCs) has been shown a significant prognostic value as a surrogate marker in various cancers. To employ CTC assay into clinical practice, a method requires rapid CTC detection in blood with high sensitivity and specificity. Here, we report a novel bioluminescence‐based assay, due to high sensitivity and extremely low background signal of bioluminescence, by genetically fusing antibody mimetics with bioluminescence proteins for CTC detection. Moreover, multi‐marker combination strategy is employed to increase detection sensitivity due to CTC heterogeneity. In the model of spiking experiments, our approach shows high specificity and sensitivity in detection of viable tumor cells with no cell toxicity observed which subsequently allows expansion of those cells in culture for CTC biobanks. Given the heterogeneous nature of CTCs, molecular characterization of CTCs at single‐cell level offers a precise investigation of patient‐specific cancer‐omics profiles for improved diagnosis and personalized treatment. To further performing post‐detection analysis of CTCs at the single‐cell level, the combination with fluorescence modality enables accurate identification of the individual spiked cells for downstream single‐cell analysis after bioluminescence detection. In conclusion, our technique represents a promising CTC detection approach for cancer management.Support or Funding InformationNational Institutes of Health (NIH)
Linking Nutrition and Molecular Biology Using Data Mining and Graph Theory
Authors: Linchangco Richard , Jay Jeremy J , Brouwer Cory 
Published: November 2023

Cardiovascular disease, cancer, and diabetes are among the leading causes of death in the United States. Factors of chronic disease include physical activity, genetics, and diet, but only physical activity and diet can be controlled. Traditionally, diet and human health have been studied through epidemiological studies at the phenotypic and observational level. The molecular mechanisms underlying the health benefits of certain foods remain largely unknown, which creates a disconnect between plant science, nutrition, human health, and molecular biology.The need to link the disparate domains of nutrition and molecular biology presents a difficult problem. Existing curated resources provide a strong baseline within the distinct domains, but data connecting them does not yet exist in machine‐readable form. This work applies data mining techniques to large collections of both medical and agricultural publication abstracts to generate links between nutrition and biology. This network of integrated data is placed into a graph database, which is the first application of Neo4j to a knowledge‐oriented nutrigenomics graph. This work enables novel queries to discover new evidence in the support of open nutrigenomics questions.Support or Funding InformationFunding from UNC General Administration and the Plant Pathways Elucidation Project
Downregulation of X‐linked inhibitor of apoptosis protein by ‘7‐Benzylidenenaltrexone maleate’ sensitizes pancreatic cancer cells to TRAIL‐induced apoptosis
Authors: Kim So‐Young , Park Sojung , Yoo Seona , Rho Jin Kyung , Jun Eun Sung , Chang Suhwan , Kim Kyung Kon , Kim Song Cheol , Kim Inki 
Published: November 2023

Tumor necrosis factor‐related apoptosis‐inducing ligand (TRAIL) is a potential biological anticancer agent. However, a wide range of human primary cancers, including pancreatic cancer, display resistance to apoptosis induction by TRAIL. Therefore, this resistance needs to be overcome to allow TRAIL to be successfully used in cancer therapy. In this study, we performed a compound screen to isolate TRAIL sensitizers and found that one of the identified compounds, 7‐benzylidenenaltrexone maleate (BNTX), sensitized pancreatic cancer cells to TRAIL‐induced apoptotic cell death. The combination of BNTX with TRAIL promoted the release of cytochrome c from mitochondria into cytosol with caspase activation and a resulting increase in annexin‐V‐stained cells. From a mechanistic perspective, we found that BNTX downregulated X‐linked inhibitor of apoptosis protein (XIAP) expression when used in combination with TRAIL and BNTX and that TRAIL‐induced apoptosis was augmented by the siRNA‐mediated knockdown of XIAP with caspase activation. We further demonstrated that BNTX promoted the ubiquitin/proteasome‐dependent degradation of XIAP protein via protein kinase C (PKC) alpha/AKT pathway inhibition. Moreover, combined treatment by BNTX with TRAIL suppressed tumor growth in vivo. Therefore, we suggest that inhibitor of apoptosis protein‐mediated resistance of pancreatic cancer cells to anticancer therapeutics can be overcome by inhibiting the PKCα/AKT pathway.Support or Funding InformationThis work was supported (40%) by (1) the Basic Science Research Program (NRF‐2016R1D1A1B03932365) funded by the Ministry of Science, Information & Communication Technology (ICT) and Future Planning, and (2) supported (30%) by a grant of the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (Grant Number: HI14C2640), and (3) supported by a Center for Women in Science, Engineering and Technology (WISET) grant (20%) funded by the Ministry of Science, ICT & Future Planning of Korea (MSIP) under the Program for Returners into R&D (KW‐2015‐PPD‐0134), and (4) supported by internal research program of ASAN Institute for Life Sciences (10%) (16‐512 & 16‐586).
Signaling Pathways Implicated in Butyrate‐Arrested Vascular Smooth Muscle Cell Proliferation
Authors: Mathew Omana P , Ranganna Kasturi , Selvam Chelliah , Yousefipour Zivar 
Published: November 2023

Prevalence and relevance of epigenetic components in cardiovascular disease pathways including those associated with atherosclerosis are now being explored to understand their impact on the pathogenesis. Butyrate, an epigenetic histone modifier with histone deacetylase (HDAC) inhibitory activity, inhibits vascular smooth muscle cells (VSMC) proliferation, an important component in the pathogenesis of atherosclerosis and alters cellular size and morphology. Since mTOR signaling pathways are crucial to cell proliferation, cell growth and survival by their multiple downstream pathways, here we investigate some of these pathways to understand the underlying mechanisms associated with butyrate effects on VSMC. Proliferating VSMC were exposed to 5 mM butyrate for different lengths of time and processed for assessing butyrate effects on the status of different signaling proteins by western analysis. Results indicate that butyrate treatment causes inhibition of mTOR expression and accordingly activation of mTOR by phosphorylation of Ser2448 is also inhibited. This inactivation of mTOR is further reflected in the inhibition of activation of its downstream effector p70S6 kinase, and in turn, inhibition of phosphorylation of p70S6 kinase substrate, S6 ribosomal protein, which are involved in the protein synthesis. Furthermore, butyrate treatment causes time‐dependent inhibition of PI3K, Akt/PKB and PDK1 activation by phosphorylation. The results indicate that inhibition of activation of both PI3K/Akt and mTOR pathways appear to be involved in butyrate arrested VSMC proliferation, which suggest that both pathways are interconnected and function as a single pathway in butyrate arrested VSMC proliferation because both pathways are crucial for the cell proliferation, cell growth and survival through the mediation of their downstream effectors.Support or Funding InformationThis study was made possible, in part, by Molecular Biology research infrastructure support from grant number 2G12MD007605 from the NIMHD/NIH”
EIF5A1 Isoform A Can Modulate the Metabolism in HeLa Cells
Authors: Pereira Karina Danielle , Tamborlin Leticia , Meneguello Leticia , Bomfim Lucas Henrique Montes , de Proença Andre Ricardo Gomes , Melo Carlos Fernando Odir Rodrigues , Catharino Rodrigo Ramos , dos Reis Silveira Leonardo , Luchessi Augusto Ducati 
Published: November 2023

The eukaryotic translation initiation factor 5A (eIF5A) is widespread in Archaea and Eukarya and contains the unusual amino acid residue hypusine. Recent studies have shown its involvement in the elongation step of translation. More specifically, acting on the synthesis of proteins containing polyproline stretches. The eIF5A has been involved in different physiological and pathological processes, such as inflammation, cancer, diabetes and virus infection. In humans, different transcript variants related to eIF5A1 are currently registered in databases. Recently, we observed that the transcript variant A is characterized by an additional upstream start codon in‐frame with the canonical initiation codon. Therefore, the product of this transcript, eIF5A1 isoform A, has a N‐terminal extension of 30 amino acid residues, when compared to the canonical isoform B. In addition it was evidenced that the isoform A connects to mitochondria. Thus, the objective of the present study is to evaluate the functionality of eIF5A1 isoform A. HeLa cells were transfected with siRNA‐VA1/2 or siRNA Control and analyzed after 72 and 96 hours. As result, it was observed that the depletion of transcript variant A correlates with reduction in cellular viability and cell cycle arrest. The metabolic profile analysis of these cells revealed that the alternative eIF5A1 isoform has some role in metabolic homeostasis. Indeed, causing negative effects on energy metabolism, as revealed by reduction in lactate production and in the content of transcripts related to mitochondria activity and biogenesis. Thus, it is proposed that eIF5A isoform A protein may modulates the cellular metabolism by acting on oxidative process controlled by mitochondria.Support or Funding InformationSão Paulo Research Foundation (FAPESP)
Metabolic Differences in Nonessential Amino Acid Plasma Concentrations in High‐ and Low‐Active Mice
Authors: Granados Jorge Z. , Ten Have Gabriella A. M. , Letsinger Ayland C. , Deutz Nicolaas E. P. , Lightfoot J. Timothy 
Published: November 2023

BACKGROUNDThere continues to be a lack of information regarding physiological mechanistic interactions between physical activity and metabolism. This is important in relation to obesity and Type II Diabetes.PURPOSETo conduct basic metabolic analysis on mice either high‐ active (HA) (C57L/J) or low‐active (LA) (C3H/HeJ), and determine basic metabolic differences between non‐essential amino acids pathways (NEAA).METHODSAll procedures were approved by the TAMU IACUC. At 12wks of age, 20 HA male mice (body weight: 27.5±1.2g; lean mass: 22.5±1.3g; fat mass: 2.5±0.7g; food intake: 3.4±0.8g) and 23 LA male mice (body weight: 25.8 ±1.2g; lean mass: 21.0±1.1g; fat mass: 2.5±0.5g; food intake: 3.0 ±1.0g) (Jackson Labs) under anesthesia had a catheter placed in the right jugular vein for blood sampling. Plasma NEAA concentrations were determined by LC‐MS/MS. Data were analyzed using unpaired t‐tests for each NEAA.RESULTSLA mice had significantly higher NEAA for GLU, ALA, CIT, ORN and for SER. Remarkable no change in GLN, ARG and GLY (Table).CONCLUSIONIn LA strain mice, specific changes in the GLU/GLN and ARG/CIT/ORN and GLY/SER pathway are observed suggesting modified pathways, linked to their lower activity levels.Support or Funding InformationStudy funded by research development grant from the Vice‐President of Research, Texas A&M University

Amino Acid
Low Active (n=20)
High Active (n=23)
Difference
P

Glutamic Acid (GLU)
36.8 ± 3.0
17.5 ± 2.0
−19.3 ± 3.6
<0.0001

Glutamine (GLN)
641.8 ± 23.9
633.7 ± 20.7
−8.0 ± 31.6
0.8011

Citrulline (CIT)
57.3 ± 2.3
46.4 ±1.5
−10.9 ± 2.8
0.0003

Serine (SER)
185.2 ± 7.1
152.3 ±5.4
−32.9 ± 8.8
0.0006

Glycine (GLY)
401.0 ± 15.4
399.0 ± 12.0
−2.1 ± 19.5
0.9167

Arginine (ARG)
104.6 ± 6.7
106.7 ± 4.2
2.1 ± 7.9
0.7895

Alanine (ALA)
671.5 ± 39.3
453.6 ± 18.7
−217.9 ±43.6
<0.0001

Ornithine (ORN)
55.4 ± 7.3
36.9 ± 2.6
−18.6 ± 7.8
0.0241

SUM NEAA
2153.6 ±105.0
1846.1 ±67.1
−307.6 ± 125.6
0.7632

Plasma concentrations in μM. Data are mean ± SE.