bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2151: Monitoring AMP Concentration,the universal product of diverse enzymatic reactions
Authors: Goueli Said A. , Hsiao Kevin , Mondal Subhanjan 
Published: August 2019

Abstract
Many enzymatic reactions use Adenosine triphosphate (ATP) as a substrate and produce adenosine monophosphate (AMP) as the common product of these reactions. AMP also functions as a modulator of many enzymes and a key cellular metabolite regulating energy homeostasis and signal transduction. For example, by activating AMP-Dependent protein kinase (AMPK), it maintains the energy charge ratio at optimal level during metabolic activities; and its concentration is altered during disregulated disease conditions. Thus, monitoring the activities of these enzymes is a primary goal for developing modulators for these enzymes. Here, we demonstrate the versatility of an enzyme coupled assay that quantifies the amount of AMP produced by any enzymatic reactions regardless of its substrates in a homogenous and high through-put formatted form. We successfully implemented it to enzyme reactions that use ATP as a substrate (aminoacyl tRNA synthetase and DNA ligase) by first depleting the remaining residual ATP substrate and converting AMP produced into ATP, the later can be quantified using luciferase/luciferin and generating light. We also tested this assay to measure direct activities of AMP generating enzymes that do not require ATP as substrate, including phosphodiesterases (cAMP) and E.coli DNA ligases (NAD+). In a further elaboration of the AMP-Glo platform, we coupled it to E.coli DNA ligase, enabling measurement of NAD+ and enzymes that use NAD+ like mono- and poly-ADP-ribosyltransferases. Sulfotransferases use 3'-phosphoadenosine-5'-phosphosulfate (PAPS) as the universal sulfo-group donor and phosphoadenosine-5'-phosphate (PAP) is the universal product. PAP can be quantified by converting PAP to AMP by a Golgi-resident PAP-specific phosphatase, IMPAD1. By coupling IMPAD1 to the AMP-Glo system we can measure the activities of sulfotransferases. In summary, we have developed a very strong platform to monitor AMP concentrations that is applicable to diverse enzymatic reactions.
Citation Format: Said A. Goueli, Kevin Hsiao, Subhanjan Mondal. Monitoring AMP Concentration,the universal product of diverse enzymatic reactions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2151.
Abstract 2159: Label-free impedance analysis as a versatile tool for dynamic screening of cancer molecular therapeutics
Authors: Cerignoli Fabio , Xi Biao , Lamarche Brandon , Zhao Leyna , Abassi Yama 
Published: August 2019

Abstract
In vitro monitoring cancer cells response to treatment often involves laborious sample processing and collects single data points that are against the dynamic nature of cancer cells. Here we present the adaptation of an impedance-based methodology to dynamically monitor cancer cell behavior and therapeutic response. The technology detects cell death, proliferation or migration by measuring changes in conductance of microelectrodes embedded in 96 and 384-wells cell culture plates. It avoids sample labeling and processing and allows continuous monitoring of cell response. Our data shows validation with cancer cell lines and primary cells monitored for cell invasion/migration through transwells or after a scratch has been performed with a dedicated device, anticancer drug response and receptor signaling activation, with similar results over end point assays. Easy experiment set up and minimal sample processing make the technology ideal for applications in large screening campaigns, while label-free technology allows less artifacts and further analysis on the same samples through orthogonal assays.
Citation Format: Fabio Cerignoli, Biao Xi, Brandon Lamarche, Leyna Zhao, Yama Abassi. Label-free impedance analysis as a versatile tool for dynamic screening of cancer molecular therapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2159.
Abstract 216: pro-Pyrrolobenzodiazepine (pro-PBD) bioconjugates, the legacy: Design and synthesis of pro-PBD conjugates containing self-immolative substituted disulfide linkers
Authors: Vlahov Iontcho , Qi Longwu , Santhapuram Hari K. , Parham Garth , Wang Kevin , Vaughn Jeremy F. , Hahn Spencer J. , Vetzel Marilynn , Nelson Melissa , Reddy Joseph A. , Leamon Christopher P. 
Published: August 2019

Abstract
Naturally occurring pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) are a group of highly potent antitumor antibiotics, isolated from various streptomycin species. PBDs covalently bond and cross-link with discrete sequences of base pairs in the minor groove of DNA. A wide range of synthetic PBDs and PBD dimers have been shown to be highly potent cytotoxic agents, and have been used as payloads in antibody drug conjugates (ADCs). However, all PBDs as stand-alone agents or as delivered via small molecule drug conjugates (SMDCs), cause undesired off-target toxicity. Recently, we designed folate conjugated pro-PBDs, masking the imine moiety with its synthetic precursors: namely, 1,3-oxazolidine carbamate and an aromatic amine. The 1,3-oxazolidine carbamate provided a conjugation site for attachment of pro-PBD to a targeting ligand. After delivery to the targeted cell and internalization, reductive cleavage of the linker system results in the generation of a 1,3-oxazolidine. The oxazolidine reacts with water to form an aldehyde which subsequently reacts intramolecularly to ultimately form the PBD diazepine ring. Herein, we report the design and synthesis of folate conjugates of pro-PBDs and self-immolative sterically hindered disulfide linker systems, which pave the way toward novel, highly-specific therapeutics which have the potential to minimize off-target toxicities.
One of the so designed conjugates, EC2629, satisfied all of our criteria for selection for pre-clinical development:
• EC2629 shows high affinity towards FR mediated binding.
• Cross-links DNA only following self-immolative release of free drug.
• Picomolar potency against FR-expressing cells.
• Curative activity with no weight loss in animal models.
• Active against drug resistant tumor xenograft models in-vivo.
• EC2629 is active against patient derived xenograft models in-vivo.
• Based on high cytotoxicity and minimal off target toxicity (in-vitro & in-vivo), EC2629 is selected for pre-clinical development.
• Biological evaluation of EC2629 was presented at World ADC, Berlin, Germany (March, 2018).
Citation Format: Iontcho Vlahov, Longwu Qi, Hari K. Santhapuram, Garth Parham, Kevin Wang, Jeremy F. Vaughn, Spencer J. Hahn, Marilynn Vetzel, Melissa Nelson, Joseph A. Reddy, Christopher P. Leamon. pro-Pyrrolobenzodiazepine (pro-PBD) bioconjugates, the legacy: Design and synthesis of pro-PBD conjugates containing self-immolative substituted disulfide linkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 216.
Abstract 2160: Microfragmented human fat tissue is a natural scaffold for drug delivery: potential application in cancer chemotherapy
Authors: Alessandri Giulio , Coccè Valentina , Pastorino Fabio , Paroni Rita , Deicas Michele , Restelli Francesco , Tremolada Carlo , Berenzi Angiola , Parati Eugenio , Brini Anna Teresa , Bondiolotti Giampietro , Pessina Augusto , Ponzoni Mirco 
Published: August 2019

Abstract
Localization of chemotherapy at the tumor site can improve therapeutic efficacy and reduce systemic toxicity. In previous studies we have shown that mesenchymal stromal cells (MSCs) isolated from bone marrow or fat tissue, can be loaded with the anti-cancer drug Paclitaxel (PTX) and kill cancer cells when localized nearby. We here investigated the capacity of human micro-fragmented adipose tissue (MFAT), used as a natural container of MSCs, to delivery PTX with the idea to improve local drug concentration and to prolong the therapeutic activity. Surprisingly, we found that both fresh but also devitalized MFAT (DMFAT) (by freezing/thawing procedure) were very effective biomaterials able to deliver and release significant amount of PTX, killing several human cancer cell lines in vitro with an impressive long lasting activity. In an orthotopic mice model of Neuroblastoma (NB) transplant, we found that DMFAT loaded with PTX prevents or delay NB relapse when placed in the surgical area of tumor resection, without any collateral toxicity. We concluded that MFAT, but also DMFAT, may work as natural biomaterials that can be used to localize and release anti-cancer molecules at the tumor site. In addition, the procedure to easily prepare DMFAT did not alter its natural scaffold structure. This suggest that such biomaterial may have a prominent role in the near future in many oncological applications to delivery anti-cancer drugs, helping to fight cancer in human.
Citation Format: Giulio Alessandri, Valentina Coccè, Fabio Pastorino, Rita Paroni, Michele Deicas, Francesco Restelli, Carlo Tremolada, Angiola Berenzi, Eugenio Parati, Anna Teresa Brini, Giampietro Bondiolotti, Augusto Pessina, Mirco Ponzoni. Microfragmented human fat tissue is a natural scaffold for drug delivery: potential application in cancer chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2160.
Abstract 2166: A novel etoposide-bound magnetic nanoparticle for remote targeting of cancer cells
Authors: Willis Alexander , Karumudi Bhargava , Liu Bing , Sough Ashley , Hwang Sangyeul , Liu Ying , Engelhard Herbert 
Published: August 2019

Abstract
Background: Magnetic nanoparticles (MNPs) have potential for enhancing drug delivery in selected cancer patients. To date, successful clinical use of MNPs has been limited due to the: 1) limited capacity for targeting at human-sized distances, 2) route of delivery, and 3) choice of therapeutic component. Here, we present data related to the creation and in vitro use of a new two-part MNP system consisting of: 1) an AuFe base particle having streptavidin binding sites, and 2) biotinylated etoposide, which was bound to the base particle. Etoposide was chosen in part due to its previous clinical use within cerebrospinal fluid, which could serve as a delivery medium for the new particle.
Methods: Base MNPs were manufactured by IMRA America, Inc., and combined with biotinylated etoposide to create AuFe-etoposide particles (etop-MNPs), which have not been previously described. Etop-MNPs were characterized by electron microscopy, UV-vis spectroscopy, and DLS. Etop-MNPs could be moved at a distance (e.g. 10 cm) by means of a “surface walking” phenomenon, occurring in response to a rotating neodymium-boron-iron permanent magnet, kindly provided by Pulse Therapeutics (St. Louis, MO). Cell lines (including D283, U138, and normal endothelial cells) were maintained using standard tissue culture technique. Viability of the cells in response to the MNP base particles, etoposide alone, and etop-MNPs, was studied by light microscopy and MTT assay.
Results: Etoposide was successfully bound to the IMRA base particle through the streptavidin-biotin interaction. Typically, 100uM etoposide was used. Etop-MNPs could be moved remotely by the rotating magnet, which produced a tangential particle velocity of 0.15 +/- 0.026 cm/sec. With 2 hours of treatment, etop-MNPs caused morphologic changes and decreased cancer cell viability to 0.50 +/- 0.03 percent that of control values.
Conclusions: These findings indicate that our novel etop-MNPs have tumoricidal effect, and can be moved relatively rapidly at physiologic distances, using a rotating magnet. Etop-MNPs, may therefore prove to be useful for magnetic drug targeting in clinical settings, such as intrathecal use within the spinal subarachnoid space.
Citation Format: Alexander Willis, Bhargava Karumudi, Bing Liu, Ashley Sough, Sangyeul Hwang, Ying Liu, Herbert Engelhard. A novel etoposide-bound magnetic nanoparticle for remote targeting of cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2166.
Abstract 2168: Testing the electrical properties of different cell lines using 3DEP reader and compare to TTFields response
Authors: Giladi Moshe , Zeevi Einav , Gotlib Karnit , Wenger Cornelia , Naveh Ariel , Bomzon Zeev , Kirson Eilon D. , Weinberg Uri , Kinzel Adrian , Palti Yoram 
Published: August 2019

Abstract
Background: Tumor Treating Fields (TTFields) are an approved anti-neoplastic treatment modality delivered via application of low intensity, intermediate frequency, alternating electric fields. The electrical properties of cells (such as permittivity and conductivity) determine the optimal frequency of TTFields that incurs the highest reduction in cell counts. Currently, there are no predictive markers for determining TTFields response or the optimal frequency to be applied for individual patient. The goal of this study is to determine the correlation between electrical properties of cells and TTFields’s optimal frequency and sensitivity. The 3DEP reader (LabTech) determines the electrical properties of cells, including permittivity and conductivity, by using Dielectrophoresis (DEP) force. DEP is a physical effect that generates a force on polarizable particles experiencing a non-homogeneous electric field and can therefore be used as a technique to analyse the way cells move within electric fields at different frequencies. Methods: The baseline electrical properties (permittivity and conductivity) of 17 cell lines from different tumor types were determined using the 3DEP reader (LabTech). The curves were analyzed using 2-way ANOVA. The optimal TTFields frequency for each cell line was determined by testing the cytotoxic effect of TTFields at various frequencies using the inovitro system. The electrical properties of cells were compared with the optimal TTFields frequency and sensitivity of each cell line. Results: The results demonstrate significant differences (p&lt;0.001) between the lower frequency range of the 3DEP curves that corresponds to membrane capacitance of cells with TTFields optimal frequency of 150kHz (9 cell lines) and cells with TTFields optimal frequency of 200kHz (8 cell lines). Membrane capacitance was also a good predictor for TTFields sensitivity based on the differences in the curves in the low frequency range. Conclusions: The results presented in this study demonstrate that cell membrane capacitance correlates with TTFields optimal frequency and sensitivity. Based on the above, there is a strong rational to further explore the potential of measuring the electrical properties of cells as a predictive marker to help determine which patient will respond better to TTFields and the optimal TTFields frequency to be applied for each patient.
Citation Format: Moshe Giladi, Einav Zeevi, Karnit Gotlib, Cornelia Wenger, Ariel Naveh, Zeev Bomzon, Eilon D. Kirson, Uri Weinberg, Adrian Kinzel, Yoram Palti. Testing the electrical properties of different cell lines using 3DEP reader and compare to TTFields response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2168.
Abstract 2172: Quantification of ERK activity in cancer cell lysates and tumor extract using differential sensing methods
Authors: Zamora-Olivares Diana , Kaoud Tamer S. , Zeng Lingyu , Telles Mitchell , Anslyn Eric V. , Dalby Kevin N. 
Published: August 2019

Abstract
The activity of protein kinases in biological samples is typically estimated by determining the phosphorylation status of either the kinase itself, or a known cellular substrate. This is typically evaluated by multi-step western blotting, or proteomics procedures. Such procedures typically provide qualitative estimates of the modifications in question. While immunoblotting and proteomics procedures can be used in a quantitative manner, their complicated workflow diminishes reproducibility. Futhermore, modifications do not necessarily correlate closely with a protein kinase’s activity. The motivation for this work was to assess the potential of a peptide array to quantify ERK activity in cancer cell lines and tumor samples, without the need to suppress related kinase activities. This work shows that a library of cross-reactive peptide-based biosensors, along with chemometric analysis can be used to profile a kinase activity in cancer cell lines. Significantly, the array is suitable for quantifying unknown levels of ERK activity in unfractionated cancer cell lysates and tumor samples using a multivariate regression model. The predicted values provided by our model were found to be comparable to those obtained by immune complex kinase assays.
Note: This abstract was not presented at the meeting.
Citation Format: Diana Zamora-Olivares, Tamer S. Kaoud, Lingyu Zeng, Mitchell Telles, Eric V. Anslyn, Kevin N. Dalby. Quantification of ERK activity in cancer cell lysates and tumor extract using differential sensing methods [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2172.
Abstract 2174: A solution for simultaneous DNA & RNA extraction from single tissue without lysate splitting
Authors: Wei Han 
Published: August 2019

Abstract
Extraction of intact and high quality gDNA or RNA is one of the most common procedures in molecular biology labs. With the advance of NGS, scientists can analyze both genomic and transcriptomic information from the same sample. Techniques that offer the ability to isolate both DNA and RNA from a single biological sample are crucial for this application. There is still great need for improved protocols in this area especially for precious and low-yield samples. Currently, most researchers use TRIzol® to isolate RNA and DNA from the same tube by extracting DNA from the organic phase and RNA from the aqueous phase. Although this method can extract high-quality RNA, the DNA yield is often low. With many biological samples in short supply, it is crucial to maximize yields. We report here using Beckman Coulter reagents, a protocol to isolate both DNA and RNA from the same fresh/frozen sample. The method utilizes size selection to separate DNA and RNA. We tested this method on 4 different frozen mouse tissues and report consistent extraction performance across all 4 tissue types. There are two advantages for this method: (1) There is no need to split a lysate for separate preparation of DNA/RNA; thus, researchers will not lose any portion of DNA or RNA. (2) Due to the different size distributions between DNA and RNA, this method offers quick and efficient extraction.
Citation Format: Han Wei. A solution for simultaneous DNA & RNA extraction from single tissue without lysate splitting [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2174.
Abstract 2176: Topical delivery of carvedilol-loaded transfersomes for photoprotection and skin cancer prevention
Authors: Chen Mengbing , Chen Mengbing 
Published: August 2019

Abstract
Ultraviolet (UV)-induced skin cancer is the most common cancer diagnosed in the United States and remains one of major public health concerns. Thus, there is an urgent need for the development of more effective strategies to prevent and treat skin cancer. We have previously demonstrated that carvedilol, an FDA approved β-blocker for cardiovascular diseases, could prevent UV-induced skin cancer both in vitro and in vivo. However, the β-adrenergic blockade effects of carvedilol might be a barrier for its repurposing as a cancer chemopreventive agent due to its cardiovascular side effects. In this work, we aimed to design a novel topical formulation, namely carvedilol-loaded transfersomes, which could maintain anti-cancer activities but avoid systemic cardiovascular effects. Carvedilol-loaded transfersomes were prepared using different phospholipids and surfactants at various ratios by a thin-film hydration method. Following optimization of preparation, the transfersomes were characterized in terms of particle size, zeta potential, and encapsulation efficiency. Skin permeation studies were performed using Franz vertical diffusion cells and skin dissected from SKH-1 hairless mice to examine the permeability of carvedilol. An optimal formulation was identified to compose carvedilol, Soy PC, and Tween-80 at a ratio of 1:3:0.5. The particle size, zeta potential and encapsulation efficiency for this formulation were determined to be 162.9±9.6 nm, 17.1±0.3 mV and 47.9±3.5%, respectively. Skin permeation studies showed that the specific carvedilol-loaded transfersomes could not penetrate through the skin layers in comparison with free carvedilol dissolved in acetone. Taken together, these results suggest that the transfersomal formulations of carvedilol may have desirable physical and biological characteristics for skin retention and local action. Ongoing studies are focused on further improvement of encapsulation efficiency and determination of its effectiveness against skin cancer.
Citation Format: Mengbing Chen, Mengbing Chen. Topical delivery of carvedilol-loaded transfersomes for photoprotection and skin cancer prevention [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2176.