bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Ras G12V/D Mutation on Downstream Pathways Using Multiplex Immunoassays
Authors: Hwang Joseph , Saporita Anthony , Xiao Qiang 
Published: June 2021

Ras proteins are members of small GTPases that function as molecular switches by alternating between inactive GDP‐bound and active GTP‐bound states. Active Ras‐GTP activate downstream pathways, including MAPK pathway by binding to Raf kinase and phosphatidylinositol 3‐kinase (PI3K) to promote cellular proliferation, survival, growth and differentiation. In the 3 decades since the discovery of RAS mutations in cancer, numerous studies have established mutant RAS as a driver of tumor initiation and progression. Oncogenic mutations in Ras are found in approximately 20% of human cancers. Furthermore, 90% of pancreatic cancers harbor K‐Ras mutations with 80% of K‐Ras mutations occurring at codon 12. RAS mutation is associated with poor prognosis due to very limited therapeutic options. To address the need for better treatment options for Ras driven cancers, the RAS Initiative was established by the National Cancer Institution. Renewed efforts in understanding Ras effectors and regulatory partners, as well as advancement in new technologies for Ras assays, are currently underway. To this end, we have developed a novel multiplex immunoassay capable of simultaneously detecting total Ras, Ras G12V, Ras G12D, phospho‐MEK1, phospho‐MEK2, B‐Raf and C‐Raf in a single well. We have tested the feasibility of using this kit on cancer derived cell lines, mouse embryonic fibroblasts, and tumor tissues. This kit was able to confirm K‐Ras G12V mutation found in the COR‐L23 cell line and N‐Ras G12D mutation found in THP‐1 cell line. Furthermore, commercially purchased colon tumor tissues characterized as Ras G12V and Ras G12D positive were confirmed using our kit. Next, we analyzed downstream pathways using commercially available immunoassay kits, such as the Multi‐pathway panel and Apoptosis panel with respect to understanding differential effect of Ras G12D vs G12V mutation. In conclusion, we have developed a novel multiplex immunoassay tool that can further our understanding of Ras mutations and its effect on downstream targets.Support or Funding InformationMilliporeSigma Corp.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of Programmed Death‐Ligand 1 in Cancer
Authors: Link Tina , Dilip Megha , Caruso Olivia , Chang Margaret , Kona Abhinav , Liu Jesse , Nathan Ajay , Singh Anita , Thomas‐Wilkinson Shiloh 
Published: June 2021

Programmed death‐1 (PD‐1) is an immunoregulatory receptor on cytotoxic T cells that suppresses the immune system upon interaction with its two ligands, programmed death ligand‐1 (PD‐L1) and programmed death ligand‐2 (PD‐L2). Programmed death‐ligand 1 (PD‐L1), previously known as “CD279”, is made up of 268 amino acids with a crystal structure. It is part of the immunoglobulin superfamily with a transmembrane domain and a tail which contains two phosphorylation sites. PD‐L1 is located on the cell surface of monocytes, B cells, T cells, and dendritic cells, and works in conjunction with PD‐1 to modulate an immune response. In binding with its ligand PD‐1 on an activated T cell, PD‐L1 suppresses the immune response of the T cell, rendering it inactive. PD‐L1's other ligand, the PD‐L2, also plays a role in deregulating the T cell immune response. Many cancer cells express PD‐L1, attenuating immunity and allowing cancer cell proliferation. This distorted relationship provides a focal point for cancer research. Attempts to inhibit the effects of PD‐L1 are concentrated on blocking its interaction to PD‐1, resurrecting the immune response, and thereby allowing CD8+ cytotoxic cells to attack tumor cells. Potential treatments include the development of an anti‐PD‐L1 antibody therapy to induce tumor remissions for patients with different types and stages of cancer. Supporting promising PD‐L1 research, the FDA has approved the PD‐L1 biomarker to accelerate investigation into a potential cure for cancer. In 2018, James P. Allison and Tasuku Hojo received the Nobel Prize in Medicine for research on PD‐L1 that lead to the discovery of checkpoint inhibitors that release immune responses, thereby reactivating T‐cells. The Walton High School MSOE Center for Biomolecular Modeling SMART Team has designed a 3‐D model of PD‐1 in interaction with its ligand PDL‐1 using JMOL to investigate the relationship between the structure and function of the proteins in relation to its current role in cancer development and its future in potential treatment.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Experimental Modal Analysis of Tumorigenesis and Cancer Metastasis
Authors: Martinez Bridget , Yang Yongchao , Farrar Charles , Mukundan Harshini , Nath Pulak , Mascareñas David 
Published: June 2021

Traditionally, performing an experimental modal analysis of a building/structure required instrumenting the structure with a spatially distributed array of accelerometers or strain gages. Alternatively, a laser doppler vibrometer would have to be scanned across the structure of interest in a sequential manner to measure structural response. Recently, researchers at LANL developed a technology that combines the theory of structural dynamics with computer vision that provides the capability to characterize structural dynamics at very high spatial density using only an imager. With this newfound success at the macro‐scale, we have exploited this novel technology to a whole new scale‐to studying the basic structure of life itself, the human cell. We hypothesize that this new technology and novel application will provide a significantly better understanding of how stiffness and mass distribution changes in a cell as it undergoes epithelial‐mesenchymal transition, and in identifying its associated EMC biochemical cues, highlight potential therapeutic targets. For the first time it should be possible to measure the high‐resolution mode shapes of cells; given that all cells undergoing cancer metastasis experience a breakdown in the cytoskeleton, this work will enable groundbreaking advances. It is imperative to highlight, that we are only beginning to understand the relationship between biophysical properties of cells and their potential to regulate tumorigenesis and motility. This knowledge could be used to provide verification and validation of finite element models of cellular structure. This work will represent the first time that expertise in experimental structural dynamics will be brought to bear on the problem of characterizing the structural dynamics of cells at high spatial resolution, which is novel and unique on its own. When successful, this new technology could be used to couple the biophysical cues associated with other detrimental human pathologies.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protease‐activated receptor 2 (PAR‐2) biased agonism in ovarian cancer progression
Authors: Pawar Nisha R. , Duru Nadire , Conway Gregory D. , Antalis Toni M. 
Published: June 2021

Ovarian cancer is one of the most fatal gynecological malignancies due to its delayed clinical presentation, indicating a need to investigate molecular mechanisms of advanced disease. Dysregulated proteolysis contributes to crucial steps of ovarian cancer metastasis, including shedding and dissemination of cells to the peritoneal cavity. Several studies have identified membrane‐anchored serine proteases, namely testisin and matriptase, that are aberrantly overexpressed in ovarian cancer. However, detailed mechanisms of activation, regulation, and downstream signaling of these proteases in this context are unknown. They have been shown to cleave and activate protease‐activated receptor‐2 (PAR‐2), a seven‐transmembrane G‐protein‐coupled receptor (GPCR). PAR‐2 has roles in hemostasis, inflammatory responses, and angiogenesis, and its overexpression has been correlated with advanced‐stage ovarian cancer. PAR‐2 has been shown to preferentially activate signaling pathways with different protease agonists, known as “biased agonism”, which can trigger different biological processes. Our overall goal is to understand and compare necessary components of testisin and matriptase activation of PAR‐2 and how resulting signaling regulates ovarian cancer progression. We hypothesize that testisin and matriptase differentially modulate PAR‐2 on the cell surface to promote cell signaling bias leading to alternative regulation of ovarian tumor progression. We are currently characterizing human ovarian cancer cell lines with stable overexpression of testisin or matriptase to compare the effects on migration, invasion, and other processes of tumor progression including metastasis using a mouse model of ovarian cancer. Interfering with these pathways might provide unique opportunities for the development of new mechanism‐based strategies for the treatment of advanced ovarian cancers.Support or Funding InformationNIH R01 CA196988, Cancer Biology T32 Training Grant (University of Maryland Baltimore School of Medicine)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biochemical Basis of Steroid Hormones Inhibition of Human Melanoma Cell Growth In‐Vitro & Its Implication on Melanoma Treatment
Authors: Ramaraj Pandurangan 
Published: June 2021

Skin, as an endocrine organ produced various hypothalamic, pituitary, adrenal and sex steroids. Steroids were not only important for healthy skin, but were also shown to influence locally and systemically. This raised the question whether melanoma was a hormone‐dependent cancer. But to the contrary, clinical study showed that menstruating females were better protected in melanoma than post‐menopausal women and men of any age. Literature showed that progesterone (P) level varied in menstruating females between 1000 to 1500 ng/dL. Whereas in post‐menopausal women, progesterone ranged between 20 to 100 ng/dL and in males between 27 – 90 ng/dL. In addition, our in‐vitro research showed that progesterone significantly inhibited human melanoma (BLM) cell growth. So, it was hypothesized that progesterone could be protecting menstruating females in melanoma. Further research showed that androgens (androstenedione, AD and testosterone, T) also inhibited human melanoma cell growth in‐vitro, but only at high concentrations (100 μM and 200 μM). Co‐incubation of AD and T separately with P showed an additive effect on BLM cell growth inhibition, suggesting the protective function of P. In order to find out the biochemical basis of steroid hormones action, Elisarray was carried out on the supernatants. There was a specific suppression of IL‐8 cytokine in the supernatants of AD, P and AD+P co‐incubated cells compared to untreated control BLM cells. Same experiments were repeated on another human melanoma (1205Lu) cell line, which also showed a specific suppression of IL‐8 cytokine. The decrease in IL‐8 was quantitated by Elisa, which correlated with the decrease in melanoma cell growth. To further confirm, experiments with T, P and T+P were carried out on BLM and 1205Lu cell lines, which also showed a specific suppression of IL‐8 by Elisarray. In addition, Kanda and Watanabe already showed that dihydrotestosterone and estrogen also suppressed IL‐8 in melanoma cells.ConclusionSuppression of IL‐8 resulting in the suppression of melanoma cell growth, implied the important role of IL‐8 in regulating melanoma growth. But suppression of IL‐8 in turn by various steroids, indicated the local regulation by the steroids. Local regulation in turn indicated the role of steroids in regulating melanoma growth. Hence, balancing or restoring local steroid hormones along with targeted or immuno therapy could be an ideal treatment for melanoma.Support or Funding Information1) Start‐up grant to PR2) Biomedical Sciences research grant3) Warner/Fermaturo research grantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Potential Block of Direct and Secondhand Smoke‐Induced Increase in Endothelin‐1 Signaling of Breast Cancer Cell Lines by Hinute‐AM
Authors: Finch Jordon , Mukherjee Shyamali , Hussein Nimo , Maebuchi Motohiro , Das Salil K 
Published: June 2021

Several epidemiological studies suggest that cigarette smoking as well as exposure to second‐hand smoke increase the incidence of breast cancer. Diet also plays an important role in breast cancer as well as other diseases. However, few studies have examined the molecular mechanisms by which cigarette smoke and diet alter breast cancer incidence. The neuropeptide hormone endothelin‐1 (ET‐1) has been identified as a multifunctional molecule that promotes cancer growth. High levels of ET‐1 expression are also associated with reduced survival of breast cancer patients. The goal of the present study was to assess whether direct or passive exposure to cigarette smoke alters the expression of ET‐1 and one of its receptors, ETAR, in non‐tumorigenic breast cells and breast cancer cell lines. To determine whether dietary factors alter this increase in ET‐1 and ETAR, cells were also exposed to Hinute‐AM, a commercially marketed soy protein hydrolysate. Proliferation assays were used to measure the effect of cigarette smoke on non‐tumorigenic breast cells. Both direct and second hand smoke exposure increased proliferation of MCF‐10, MCF‐12A and MCF‐12F non‐tumorigenic breast cell lines. We next performed Western blot analysis to test the effect of cigarette smoke of ET‐1 and ETAR expression. Exposure to direct and second‐hand cigarette smoke increased ET‐1 and ETAR protein within non‐tumorigenic MCF‐10A cells as well as BT549 and MCF‐7 breast cancer cells. Low concentrations (0.02–0.20 mg/mL) of Hinute – AM reduced proliferation of breast cancer cells and protein expression of ET‐1. In conclusion, our data suggest that cigarette smoke may increase the proliferation and aggressiveness of breast cancer by increasing expression of ET‐1 and its receptors. Furthermore, it may be possible to counteract the effects of cigarette smoke with dietary supplements such as Hinute‐AM.Support or Funding Information[Supported by Grants from Army (DAMD 17‐03‐1‐0352), FAMRI (Clinical Innovator Award 062415), Fuji Oil Company, Osaka, Japan, NCI (R25CA214220) and NIH (MeTRC5U5AMD007593).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluating the impact of bead media diameter and material composition on bacterial cell lysis and genomic DNA extraction
Authors: Proctor Caleb , Soldat Shelby M , Easparro Brandon , Nash Rodney , Atwood James 
Published: June 2021

Mechanical lysis of bacteria is a common alternative to enzymatic, detergent, or tonicity‐based methods. Mechanical lysis is needed in areas of research where introducing chemicals can compromise either the assay or the analyte and is achieved in a variety of ways, one of which is bead beating. Bead beating utilizes small beads of various sizes and composition to contact the sample with high energy to facilitate lysis. This type of lysis can be achieved using many different types of equipment from laboratory vortexers to specialized bead mills. Maximizing the amount of lysis is important for both consistency of analyte extractions and increasing yield when sample size is limited.Herein, we evaluate two bead media compositions (borosilicate glass and yttria‐stabilized zirconium oxide) and two sizes of each of those compositions (0.5 mm and 0.1 mm) to analyze what factors are most important to achieve the maximum amount of lysis using cultures of Escherichia coli, Staphylococcus epidermidis, and Nocardia brasiliensis. Speed, time, number of beads, and mass were all controlled to identify the ideal size, composition, force and duration to maximize cell lysis while maintaining nucleic acid integrity. In order to evaluate lysis efficiency cells were disrupted then plated to enable colony counting. After each processing step, genomic DNA was purified, quantified, and analyzed by gel electrophoresis to evaluate genomic DNA integrity.In all cases, it was determined that bead diameter played a greater role in lysis efficiency when compared to bead material even though kinetic energy of the zirconium oxide beads is 2X greater than the glass at a given tube velocity. E. coli was lysed at greater than 90% total cell lysis in three minutes at a tube velocity of 4.2 m/s (Figure 1). No DNA degradation was observed after homogenization for 1 minute. Increasing amounts of DNA shearing was observed as processing was increased (Figure 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.
Symbiosis and Host Immunity in Reef Ecosystems
Authors: Lazar Stephen , Palmer Andrew G 
Published: June 2021

Many marine reef‐building corals live in nutrient poor conditions, forming symbiotic relationships with dinoflagellates of the genus Symbiodinium to supplement or replace active feeding. In order to maintain this symbiotic state, and avoid being consumed or expelled, these symbionts suppress the immune system of the host. Yet these corals do not show high levels of disease under standard conditions. The simplest hypothesis is that Symbiodinium are able to compensate for this reduction in immunity. Many common reef diseases are associated with bacteria which regulate the transition from being benign to virulent through the population detecting process of quorum sensing (QS). Other algae have proven capable of metabolizing the N‐acyl‐L‐homoserine lactone (AHL) class of QS signals, potentially disrupting these events. In this study we have investigated the ability of members of the genus Symbiodinium to regulate bacterial quorum sensing as a strategy for supplementing the host immune system.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synthesis and Biological Evaluation of a Photoswitchable Quorum Sensing Molecule
Authors: Karns Taylor , Stajdl Shelby , Zimmer Kassidy , Rohlman Christopher , Streu Craig 
Published: June 2021

Quorum sensing is the ability of bacteria to communicate with one another via chemical signals. This ignites a response within the collective group of bacteria to produce a single response that ultimately benefits the bacteria, an example being biofilm formation. If this natural process could be controlled or coopted, the biotecnical applications arising from this natural ability of bacteria could be limitless. Photoswitching molecules are able to respond to a light stimiulus. Azostilbenes in particular isomerize from trans to the cis conformation in response to the appropriate wavelengths of light. If one of the available conformations is bioactive, while the other is not, this photoisomerization has the effect of activating or inactivating the molecule in response to light. The power of light responsive chemical signaling systems comes from their ability to achieve exquisite spatial and temporal control, which is of tremendous utility for numerous bioindustrial applications. To this end, we have designed a series of photoswitching molecules based upon N‐acyl homoserine lactones (AHLs), which are known to be prominent chemical messengers in numerous bacterial species, by integrating the azo‐stilbenoid motif into the parent molecule scaffold. These molecules are made by Mills coupling between the appropriate aniline and aryl‐nitroso species. Their photokinetic properties are measured by UV/Vis spectroscopy and their bioactivity measured in the presence and absence of isomerizing light. We herein describe the synthesis, photokinetic studies, and bioactivity of this class of photoswitchable molecule. We also describe potential industrial applications of this technology.Support or Funding InformationAlbion College Foundation for Undergraduate Research, Scholarship, and Creative ActivityThis 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 Prostate Cancer Cell Proliferation and Survival by Rosemary Extract
Authors: Jaglanian Alina , Tsiani Evangelia 
Published: June 2021

Prostate cancer is the most commonly diagnosed cancer in men. Despite many advances in diagnosis and treatment, prostate cancer is the second‐leading cause of cancer related death amongst men in North America and therefore there is an urgent need to find novel chemicals that can be used effectively towards the treatment of prostate cancer patients. Prostate cancers are typically characterized by the expression of androgen receptor (AR) and prostate‐specific antigen (PSA). Prostate cancers that are AR positive can be treated with hormonal therapy, however those which are AR negative are more aggressive and do not respond to hormone therapy, thus this subtype of prostate cancer requires targeted treatments. Many chemotherapeutic drugs have been discovered by screening natural products from plants. Polyphenols derived from the plant rosemary, have been found to have antioxidant and anti‐inflammatory effects and are proposed to have anticancer activity. In previous studies, rosemary extract (RE) has been found to reduce cell proliferation, viability, survival, and increase apoptosis in various cancer cell lines. However, there is very limited evidence examining RE's effects in androgen receptor negative prostate cancer cells. In the present study, we examine the effects of rosemary extract on PC‐3 prostate cancer cell proliferation, survival, migration and apoptosis. RE significantly inhibited PC‐3 cell proliferation and survival in a dose dependent manner. Furthermore, RE reduced cell migration and enhanced the cleavage of PARP, a marker of apoptosis. Our findings indicate that RE may have potent anti‐cancer properties against AR negative prostate cancer and supports the need for further investigation into its role in modulating key signaling pathways involved in cell proliferation, survival, and migration.Support or Funding InformationSupported by the Prostate Cancer Fight Foundation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.