bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of choline acetyltransferase activity abrogates the growth of lung adenocarcinoma patients who are exposed to tobacco smoke
Authors: Akers Austin T. , Brown Kathleen C. , Colclough Kate W. , Nolan Nicholas A. , Friedman Jamie R. , Miles Sarah L. , Bow Eric W. , Rimoldi John M. , Hardman Elaine W. , Dasgupta Piyali 
Published: June 2021

The pathophysiological characteristics of lung adenocarcinoma (LAC) in smokers is divergent from LAC in non‐smokers. One of the distinctive features of LAC in smokers is that the disease is relatively resistant to targeted molecular therapies. For example, targeted therapeutic agents like EGFR‐inhibitors (erlotinib and gefitinib) are highly effective in LACs in non‐smokers. However, these agents display much lower anti‐tumor activity in LACs in patients who are active smokers. Similarly, lung cancer patients (who are active smokers) show a lower response to chemotherapy than those who are non‐smokers. However, the majority of LAC patients are smokers. This underlines the need to identify viable drug targets for LAC therapy in patients who are exposed to cigarette smoke. A survey of literature reveals 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 the mitogenic activity of nicotine is that it promotes the production 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 analogous to their 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. The anti‐tumor activity of BW813U is also observed in H838 cells (belonging to an 80 pack‐year smoker) xenografted on athymic mice. Our studies show that antagonists of ChAT like BW813U may have therapeutic applications in the majority of the population of LAC who are smokers.Support or Funding InformationFunding for our study was supported by a NIH R15‐AREA Grant (2R15CA161491‐02). 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).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Is HPV Vaccination Secure Protection of Women Without or With Cytologic Cancer Screening
Authors: Markovic Olivera , Markovic Nenad 
Published: June 2021

Today, million of girls around the world receive vaccination against HPV hoping vaccines will protect them from the risk of getting cervical cancer if having sex with HPV positive partners.But, is it true? And, if it is true, how long the protection lasts?This dilemma has controversial documented data. Therefore, manufacturer of HPV vaccines and health care providers in charge of cervical cancer prevention, advertised, via guidelines, testing for antibody titer and for cytopathological changes of cervical specimen – similar to Pap test. The open dilemma is about scheduling: from 1‐to 5‐year periods between screening. Many studies have been published and supported with statistical data, but, indeed, this is only a guessing game.The Pap test was introduced as screening for lesions which can be developed into cervical cancer and the suspect lesions were recommended for colposcopy/biopsy/histology – finally to early removal of those lesions. This removal of lesions, not the Pap test alone, has interrupted the natural history of cervical cancer. The definition “cancer is unstoppable growth of malignant cells,” was replaced with “no lesion – no growth of cancer.”HPV vaccine protection is specific for few strains of HPV virus which have been found in developed cervical cancers. Cervical cancer has a much wider palette of risk factors including other HPV strains, and other promoters of malignancy [3]. Because of that, the cytopathologic screening must be kept regularly active. This fact returns us to the old concerns: Pap test staining has inherited 20% false negative readings. The recommended MEDYKO composite biomarker [1–3] consists of metabolic biomarker, “ME” (increasing sensitivity to include BCC category), “DY” dysplastic changes defining the level of clinical diagnosis, and “KO” (koilocytes, HPV disease indicating worse prognosis).All three markers are visible on one slide reducing false negative results of Pap test, and false positives of HPV infection testing. All are amenable for digital communication between POC and remote medical centers. Further, the specimen can be obtained at home with the Self‐sampling Kit what increases the outreach and opens the opportunity for annual testing.The concept has been tested, confirmed, but still, waits for mass applications.ConclusionCytologicl testing is still necessary for HPV vaccinated women, but Pap tests needs improvement as MEDYKO™ described above, annual home specimen sampling, collection of specimens in scattered POC, evaluation in remote specialist centers and early, the same day, intervention for removal of lesions if necessary.Support or Funding InformationNo external supportThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic cadmium exposure reduces the dependence of MCF7 cells on ERa for cell growth
Authors: Bloomfield Mathew , Louie Maggie 
Published: June 2021

The global prevalence of breast cancer in women illustrates the importance of identifying factors that contribute to disease onset and progression. Endogenous and environmental agents that interact with estrogen receptor alpha (ERa) have been shown to play a role in breast cancer etiology. Evidence from epidemiological studies and animal models have suggested that cadmium, a heavy metal that can activate ERa, contributes to the development and progression of breast cancer. Additionally, our lab showed that chronic cadmium exposure altered the expression of several ERa‐responsive genes and increased the malignancy of MCF7 breast cancer cells. Although these studies support cadmium's function as a hormone disrupter, the role of ERa in cadmium‐induced breast cancer progression remains unclear. Therefore, we modulated the expression of ERa in MCF7 cells after chronic cadmium exposure (Cd7 and Cd12) in order to understand its role in cadmium‐induced cancer cell growth and gene expression. The doubling times of MCF7, Cd7, and Cd12 cells were increased as a result of ERa loss, but this effect was significantly less in the cadmium‐adapted cells suggesting chronic cadmium exposure reduces the dependence of MCF7 cells on ERa for cell growth. Furthermore, preliminary analysis showed the transcript levels of classical and non‐classical ERa‐regulated genes were reduced in MCF7 cells after transient and permanent modification of ERa expression, while the non‐classical targets were not as affected in Cd7 and Cd12 cells after ERa knockout indicating cadmium exposure may have altered the regulation of these genes. These results demonstrate that while ERa still contributes to the growth of MCF7 cells chronically exposed to cadmium, their dependency on ERa appears to have diminished. The relevant processes affected by chronic cadmium exposure remain unclear, but these may enable MCF7 cells to adapt to ERa loss and potentially provide a path to ERa independence as the cancer progresses.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Study of the Chemo‐sensitizing Effects of Thymoquinone, the Active Constituent of Negilla Sativa (Black Seed), to Nitrogen Mustards, Chlorambucil and Cyclophosphamide, in Human Melanoma Cell Lines
Authors: Alsulaimany Marwa , Kerr Stephen G. 
Published: June 2021

Black seed is often used as a spice in food and as a preservative in many products. The main active constituent of black seed oil is thymoquinone (TQ) which constitutes 30–57% of the oil. Thymoquinone has shown varying therapeutic benefits in various disease states, including cancer. The aim of this study was to investigate the cytotoxic and chemo‐sensitizing potential of TQ and compare it to ethacrynic acid (EA), a known cytotoxic agent in sensitizing melanoma cells to the effects of the N‐mustards, chlorambucil (CHL) and cyclophosphamide (CP). Both TQ and EA have an α, β unsaturated carbonyl system which could act as potent Michael Acceptors to mediate the cytotoxicity. Studies also assessed the cytotoxic potential of cyclophosphamide(CP) and chlorambucil (CHL) alone and in combination with TQ. Approximately 1×104 cells/100μL/well of two melanoma cell lines, lightly pigmented SK‐MEL19 and pigmented SK‐MEL23 were seeded in 96‐well plates and incubated for 24h at 37C° and 5% CO2. The cells were then treated with TQ (25–500 μM), EA (25–200 μM), CHL (25–500 μM) and CP (25–500 μM), for 24h and 48h. Combination treatment included pretreatment of the cells with TQ (100 μM) for 24 h followed by further exposure to CHL for an additional 24 h. Similarly, cells were pretreated with EA (100 μM) for 24h, followed by further exposure to CHL for 24h. Colorimetric MTS (Abcam) cytotoxicity assays, per manufacturer's instructions, was then carried out to determine cell viability. The results indicated that TQ exerts its toxic effect in a concentration‐dependent manner in SKMEL19 cell lines at concentrations ≥75μM, while in SK‐MEL23 at concentrations ≥ 100 μM, relative to the control. In addition, EA reduced cells viability significantly in both cell lines at concentration of 100 μM and above. However, CP did not show any cytotoxic effect in both cell lines at 24h or 48h up to concentrations of 500 μM. In contrast, CHL reduced cell viability in both cell lines at high concentrations (250 and 500μM) within 24h. When the incubation time was increased to 48h, the cytotoxic effect of CHL in SK‐MEL19 was observed at concentrations ≥ 100 μM, while in SK‐MEL23 at concentrations ≥ 150 μM. Pre‐treatment with TQ (100 μM), as well as EA (100 μM) greatly enhanced the cytotoxic effect of a non‐toxic dose of CHL (100 μM) in both cell lines after 24h as compared to the effect of CHL alone. These studies demonstrate the potentiating effect of TQ on CHL cytotoxicity possibly by sensitizing the human melanoma cell lines to the chemotherapeutic agent.Support or Funding InformationMA acknowledges SACM for financial supportThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determination of 13 ginsenosides in diverse ginseng product by LC‐MS/MS
Authors: Cho Pil Joung , Gao Yan , Jo Jung Jae , Lee Sangkyu 
Published: June 2021

Ginseng (Panax ginseng Meyer) has been used as traditional herbal drug in Asian countries. Ginsenosides, the ginseng saponins, are major components, have pharmacological and biological efficacy, including anti‐inflammatory, anti‐diabetic and anti‐tumor effects. To control the quality of the components in diverse ginseng products, we developed a new quantitative method using LC‐MS/MS for 13 ginsenosides: Rb1, Rb2, Rc, Rd, Re, Rf, 20(S)‐Rh1, 20(S)‐Rh2, Rg1, 20(S)‐Rg3, F1, F2, and compound K. The method was successfully validated and applied in four representative ginseng products. This quantification method was applied to several processed ginseng products including fresh ginseng powder (FGP), white ginseng powder (WGP), red ginseng powder (WGP), and red ginseng concentrates (RGC). The amounts of the 13 ginsenosides in the various type of ginseng samples could be analyzed simultaneously. This method was expected to be suitable for quality control of ginseng productsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MICROINJECTION OF UROTENSIN II INTO THE PEDUNCULOPONTINE TEGMENTNUM LEADS TO AN INCREASED PERCEPTION OF SENSORY STIMULI
Authors: Ettaro Robert , Daniels Derek , MacLaren Duncan , Markovic Tamara , Clark Stewart 
Published: June 2021

The pedunculopontine tegmentum (PPTg) plays a role in processing multiple sensory inputs and innervates brain regions associated with reward‐related behaviors. The urotensin II receptor, activated by the urotensin II peptide (UII), is selectively expressed by the cholinergic neurons of the PPTg. Although the exact function of cholinergic neurons of the PPTg are unknown, they are thought to modify the perception of reward magnitude or salience detection. We hypothesized that the activation of PPTg cholinergic neurons would alter sensory processing across multiple modalities (ex. taste and hearing). Here we had three aims: first, determine if cholinergic activation is involved in consumption behavior. Second, if so, distinguish the impact of the caloric value by using saccharin, a zero calorie sweetener. Lastly, we tested the perception of acoustic stimuli by measuring the acoustic startle reflex (ASR).Male Sprague‐Dawley rats were bilaterally cannulated into the PPTg, then housed in a contact lickometer and placed under food restriction lasting the entire consumption experiment (water ad lib.). The lickometer recorded each individual lick to give insight into the microstructure of the consumption behavior. Treatment consisted of a microinjection of either 1 μL of aCSF or 1 μL of 10 μM UII into the PPTg, and the rats were immediately given access to either sucrose or saccharin. For the remaining five days rats were allowed one hour access per day to the same sweet solution without any further treatments. ASR testing consisted of a baseline (no treatment), treatment day, and two additional days (no treatment). Immediately following the microinjection of UII, consumption of both saccharin and sucrose increased compared to controls. The significant difference in sucrose consumption lasted two days post‐treatment while saccharin consumption was significantly different for three days after treatment. The ASR was significantly increased the day after treatment in the rats treated with UII.Activating cholinergic PPTg neurons may lead to a miscalculation of the salience of external stimuli, implicating the importance of cholinergic input in modulating a variety of behaviors. This potential plastic change can give rise to further studies in the role of sensory processing on reward related‐behaviors at the level of the PPTg cholinergic neurons.Support or Funding InformationThe Howard T. Blane Director's Award in the Addictions (BDAA) from Research Institute on AddictionThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Adult Neural Stem Cells Show Regional and Sex‐Dependent Responses to Chronic Poly‐Drug Administration
Authors: Schlagal Caitlin , McGrath Erica , Gao Junling , Dunn Tiffany , Fox Robert , Stutz Sonja , Dineley Kelly , Kaphalia Bhupendra , Cunningham Kathryn , Wu Ping 
Published: June 2021

Cocaine use disorder (CUD) and alcohol use disorder (AUD) are highly prevalent substance use disorders in the US. Cocaine and alcohol co‐abuse is the third most lethal substance combination and affects 12 million Americans annually. Interestingly, it has been reported that females experience greater euphoria when cocaine and alcohol are combined. Individual use of cocaine and alcohol cause neurodegeneration; however little is known about the combined effects of these substances on the neural stem cell (NSC) population. NSCs are a population of cells in the brain maintained throughout life that are characterized by their ability to self‐renew and give rise to new neurons and astrocytes. Efforts in drug addiction research primarily focus on preventing or stopping abuse, however little work is being done to reverse brain damage incurred by chronic drug abuse. Therefore, the NSC population represents a potential therapeutic target due to their regenerative capabilities. The aim of our study is to evaluate regional and sex differences of endogenous adult neural stem cells to chronic treatment with alcohol and/or cocaine. We sought to elucidate the response of adult endogenous NSCs to chronic alcohol and cocaine treatment using an inducible lineage tracing mouse model. Adult mice were randomly divided into 1 of 4 groups: control, cocaine, ethanol, or combination treatment. Daily i.p. injections of cocaine were administered and ethanol was provided in a complete nutrient liquid diet to appropriate groups. Brain tissue was analyzed for markers of NSC survival and differentiation in two distinct brain regions, the subventricular zone of lateral ventricle (SVZ), and subgranular zone of dentate gyrus (SGZ). The SGZ was also evaluated for changes in metabolic enzymes associated with ethanol and cocaine metabolism. We found that NSCs have a unique response to drug depending on the regional location and sex of the animal. Females had more robust decreases in neural stem cell (NSC) survival in the SVZ compared to males. SGZ neurogenesis was reduced drastically in the combination group. There were significant increases in metabolic enzyme expression in response to drug treatment, with notable differences between sexes. Overall, we show drastic changes to the NSC population following exposure to cocaine and ethanol, as well as alterations in metabolic enzyme expression.Support or Funding InformationJohn S. Dunn Foundation NIDA T32 Award (Grant# 5T32DA007287‐17 and 3T32DA007287‐18S1)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Desensitization of PGE2‐stimulated cAMP Signaling Due to Upregulated Phosphodiesterase Activity in Human Lung Fibroblasts
Authors: Nuñez Francisco , Corpuz Maia , Kazarian Austin , Johnstone Timothy , Ostrom Rennolds 
Published: June 2021

Pulmonary fibroblasts play an essential role in maintaining the structure and function of the lung. In idiopathic pulmonary fibrosis, fibroblast cells persist in an activated form and produce excessive fibrous material, leading to loss of alveolar structure. PGE2, which specifically activates EP2 and EP4 receptor subtypes, is a cAMP elevating agent and thus has anti‐fibrotic potential, since cAMP has been shown to have anti‐fibrotic properties. Although PGE2 levels are increased in the bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis (and in animal models of pulmonary fibrosis), it does not prevent pulmonary fibrosis. One hypothesis is that PGE2 effects are self‐limiting because of desensitization caused by prolonged exposure. To test this hypothesis, we pretreated HFL‐1 fibroblasts with PGE2 and other cAMP‐elevating agents then measured PGE2‐stimulated cAMP levels using the downward cAMP difference detector in situ (cADDis) cAMP sensor (Montana Molecular, Bozeman MT). After 24‐hour pretreatment with 100 nM PGE2, we changed the media and measured cAMP kinetics in response to various concentrations of PGE2. Pretreatment with PGE2 shifted the concentration‐response curve of PGE2 rightward approximately 50‐fold. Neither 100 nM isoproterenol or 1μM forskolin pretreatment for 24 hours altered PGE2 response, implying that other cAMP elevating agents do not induce desensitization. We sought to uncover the underlying mechanism responsible for PGE2 desensitization. Desensitization could result from either receptor internalization through the GRK/β‐arrestin mediated pathway or via increase PDE activity. To determine if PGE2 pretreatment increases PDE activity, we used the PDE3 inhibitor, cilostazol, and the PDE4 inhibitor, rolipram after PGE2 pretreatment. Rolipram, increased both the EC50 and Emax of PGE2 in vehicle pretreated HFL‐1 fibroblasts. In cells pretreated with PGE2, rolipram partially reversed the desensitization, indicating that PDE4 may be induced by long term PGE2 exposure. Cilostazol had minimal effects on either control or desensitized PGE2 responses, implying that PDE3 is not involved. Examination of PDE isoform mRNA levels via quantitative RT‐PCR, show that PGE2 treatment upregulates PDE3A, PDE4C and PDE4D, while pretreatment with isoproterenol or forskolin only upregulated PDE4D. Taken together, these results show that long‐term exposure to PGE2 causes desensitization of EP receptor‐stimulated cAMP signaling, partly through the increased expression of a PDE4 isoform. Defining the precise PDE isoform involved in the desensitization of PGE2 responses may lead to new therapeutic strategies for treating pulmonary fibrosis.Support or Funding InformationThis work was supported by NIH grant GM107094 (RO)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.