bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structure Activity Relationship Study of CAPE and its Derivatives on Liver Carcinoma
Authors: Nguyen Julie , Bhave Vishakha , Hielscher Abigail , Wang Xinyu 
Published: June 2021

Globally, the sixth most common cancer and the second largest contributor to cancer mortality is malignancy of the liver. The most frequent form of liver cancer is hepatocellular carcinoma (HCC), a primary liver cancer of the parenchymal cells. Presently, there are no effective treatment options aside from liver resection and transplantation. For this reason, caffeic acid phenethyl ester (CAPE) has been proposed as a potential therapeutic candidate. CAPE, a phenolic compound found in bee propolis, is reported to have a plethora of bioactive properties including antitumor traits. The objective of this study is to elucidate the relationships between the structure of CAPE and its effect on liver cancer cell growth in addition to exploring the potential mechanism for this. Seven CAPE derivatives were examined including one novel derivative previously synthesized in our lab, fluorinated caffeic acid phenethyl ester (FCAPE). We evaluated the effect of CAPE and its derivatives on the viability and proliferation of a human HCC cell line, HepG2. Our preliminary data show that CAPE and some of its derivatives reduce the viability and inhibit the proliferation of HepG2 cells in a dose‐ and time‐dependent manner. Comparison of CAPE derivatives' structures versus their activity indicates that the catechol ring functional group is required for their inhibitory effect on HepG2 cell growth. To examine whether induction of apoptosis is responsible for the cell death, western blot was conducted to check protein expression of apoptosis‐related factors including caspase‐3 and poly‐(ADP‐ribose) polymerase (PARP). Our data show the breakdown of caspase‐3 and PARP by CAPE, suggesting that CAPE may induce apoptosis of HepG2 cells. In conclusion, our data suggest that CAPE inhibits the growth of HepG2 cells through induction of apoptosis, and this effect of CAPE requires the integrity of a catechol ring functional group in order to carry out its cytotoxic effects.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anticancer Activity of A Silyl Andrographolide Analogue Mediated Through Wnt/β‐Catenin Signaling In Colon Cancer Cells
Authors: Reabroi Somrudee , Saeeng Rungnapha , Chairoungdua Arthit , Piyachaturawat Pawinee 
Published: June 2021

Hyperactivation of Wnt/β‐catenin signaling implicated in oncogenesis of colorectal cancer (CRC) is a potential molecular target for chemotherapy. An andrographolide analogue, 3A.1 (19‐tert‐butyldiphenylsilyl‐8, 17‐epoxy andrographolide) has previously been reported to be potently cytotoxic toward cancer cells by unknown molecular mechanisms. The present study explored the anti‐cancer activity of analogue 3A.1 on Wnt/β‐catenin signaling in colon cancer cells (HT29 cells) which were more sensitive to the others (HCT116 and SW480 cells). Analogue 3A.1 inhibited viability of HT29 cells with IC50 value of 11.1 ± 1.4 μM at 24 h, which was more potent than that of the parent andrographolide. Analogue 3A.1 also suppressed the proliferation of HT29 cells and induced cell apoptosis in a dose‐dependent manner. Its apoptotic activity was accompanied with increased expressions of proteins related to DNA damages; PARP‐1 and γ‐H2AX. In addition, analogue 3A.1 significantly inhibited T‐cell factor and lymphoid enhancer factor (TCF/LEF) promoter activity of Wnt/β‐catenin signaling. Accordingly, the expressions of Wnt target genes and β‐catenin protein were suppressed. Moreover, analogue 3A.1 increased the activity of GSK‐3β kinase, which is a negative regulator responsible for degradation of intracellular β‐catenin. This mode of action was further supported by the absence of the effects after treatment with a GSK‐3β inhibitor, and over‐expression of a mutant β‐catenin (S33Y). Our findings reveal, for the first time, an insight into the molecular mechanism of the anti‐cancer activity of analogue 3A.1 through the inhibition of Wnt/β‐catenin/GSK‐3β pathway and provide a therapeutic potential of the andrographolide analogue 3A.1 in CRC treatment.Support or Funding InformationThis research project was supported by TRF through the RGJ‐Ph.D. Program (PHD/0193/2553 to PP and SR) and the research grant IRN‐58W0004.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A unique method for antibody to possess the catalytic function (3rd report)
Authors: Uda Taizo , Akiyoshi Yuko , Taguchi Hiroaki , Hifumi Emi 
Published: June 2021

Since a natural catalytic antibody found at 1989, many mouse‐ and human‐type of catalytic antibodies cleaving the targeting antigens such as peptides, nucleotides, viruses and bacterial proteins, molecules such as factor VIII, CCR5, TNFα etc. have been produced so far (1, 2). In contrast, thousands of monoclonal antibodies (mAbs) have been produced over past 40 years. However, not all monoclonal antibodies have catalytic activity. It is considered that the proportion possessing a catalytic function is about several percentages or lower. Thus, the authors have continued to explore the technology how to put the catalytic function to those normal (non‐catalytic) mAbs. Recently, we have made a significant progress in part with respect to the technology.After the genes of human antibody light chain were inserted into pET20b (+), E. coli was transformed and each gene was expressed by inducing with IPTG. After recovering the soluble fraction of the culture supernatant, it was purified by Ni‐NTA‐, cation‐, and/or size exclusion‐chromatography. The enzyme activity was measured using Arg(R)‐pNA as a synthetic substrate and/or the antigens such as amyloid‐beta (Aβ) peptide etc. SinceWe found interesting two light chains, #7TR and #7GY. The former could degrade both FRET‐Aβ substrate and Aβ1–40 full peptide. In contrast, #7GY, whose sequence is identical to that of #7TR except for the amino acids of the positions at 29th and 30th, did not degrade the FRET‐Aβ substrate and Aβ1–40 full peptide at all. The chemical features of the two light chains were investigated in detail by using a synthetic substrate, Arg‐pNA. #7TR showed higher hydrolytic activity for Arg‐pNA substrate than that of #7GY by a factor of one and half. In addition, the presence of Zn (II) ion hugely enhanced the catalytic activity of #7TR light chain but not #7GY. Fluorescence spectroscopy clearly exhibited the difference of the two antibody light chains. Through these facts and discussion, we will propose a clue how to we can put the catalytic function to a normal (non‐catalytic) antibody.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis of the food components and physiological activities of Brassica juncea from Jeongseon
Authors: Sim Wan‐Sup , Choi Sun‐Il , Jung Tae‐Dong , Cho Bong‐Yeon , Choi Seung‐Hyun , Han Xionggao , Kim Young‐Cheul , Lee Ok‐Hwan 
Published: June 2021

This study was conducted to provide basic data of Brassica juncea from Jeongseon (BJJ) which is the food components and physiological activities. The moisture, crude protein, crude fat, crude ash and carbohydrate content of BJJ were 91.58±0.08%, 2.29±0.26%, 0.07±0.00%, 1.17±0.00% and 4.89±0.03%, respectively. The sugar content was 0.29±0.02 g/100 g of glucose and 0.10±0.00 g/100 g of fructose. Major fatty acids were palmitic acid(C16:0), octadecenoic acid(C18:1n9c) and stearic acid(C18:0), especially the composition of the palmitic acid was the highest at 31.22%. Major minerals were K, Ca, P, Na, Mg, especially the K content was highest at 464.31±6.33 mg/100 g. Vitamin was mainly composed of Vitamin B1, B2, B6, C, E. The primary organic acid was 305.26±0.90 mg/100 g of acetic acid. Total amino acid was mainly composed of L‐Proline at 774.30±119.06 mg/100 g. DPPH radical scavenging activity and FRAP value increased depending upon the concentrations of BJJ extract (0.1, 1.0, 5.0 mg/mL). Total phenolic contents and total flavonoid contents were 7.88±0.06 mg GAE/g, 37.31±0.39 mg RE/g. In addition, we assessed the lipid accumulation and the antioxidant effect of BJJ extract on 3T3‐L1 and Human Dermal Fibroblasts (HDFs), respectively. After we confirmed no significant difference of cell viability between control group and BJJ extract of 25, 50, 100 μg/mL through XTT assay, we applied this concentration of BJJ extract on the continuous experiments. Oil Red O (ORO) staining showed that the lipid accumulation was inhibited by 25% at a concentration of 100 μg/mL compared to control. Through NBT assay, the reactive oxygen species (ROS) production decreased in all experiment groups treated by BJJ extract, especially the ROS production decreased about 23% at a concentration of 100 μg/mL compared to control. However, at low concentrations (25, 50 μg/mL), the protective effect of BJJ extract was no significant effect but 100 μg/mL. These results suggested BJJ has a potential as a functional food material.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Coupling Fluorescence‐Activated Cell Sorting and Targeted LC‐MS/MS for Epi‐Proteomic Analysis of Normal Leukocytes
Authors: Camarillo Jeannie , Swaminathan Suchitra , Abshiru Nebiyu , Sikora Jacek , Morris Juliette , Kelleher Neil , Thomas Paul 
Published: June 2021

Fluorescence‐activated cell sorting (FACS) is widely used to isolate subtypes of various immune cells from human blood. Most recently, these subpopulations have been subjected to next generation sequencing to elucidate changes in gene expression associated with differentiation. Histone modifications are essential for regulating chromatin and maintaining gene expression, and few studies have investigated global alterations in histone modifications required for differentiation. Here, we describe an approach for integrating FACS and targeted mass spectrometry to define a global “epiproteomic” signature for white blood cells from normal individuals. Using our multiple‐reaction monitoring LC‐MS/MS method, which can measure 90+ histone modifications on all core histones, we assessed changes in histone modifications from different collection methods and cell types. First, collection buffer conditions were optimized to allow for sorting directly into histone extraction buffer, thereby reducing the likelihood of changes in histone modifications that could occur during long sorts. Next, we sorted neutrophils, monocytes, B cells, and T cells and identified differences in histones modifications across these broad cell types. Finally, FACS was used to sort naïve, TH1, and TH2 T cells from human peripheral blood for histone analysis, revealing little differences in histone marks in these closely‐related subtypes. This method allows for interrogation of global changes in histone modifications in distinct cell types, defining an “epi‐proteomic” signature that can be used to more fully understand differentiation in hematopoiesis and disease development.Support or Funding InformationThis work was carried out with financial support from the National Resource for Translational and Developmental Proteomics (P41GM108569).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Revisiting the Pharyngeal Apparatus: Arches, Pouches, & Grooves—Oh My!
Authors: Dennis Jennifer 
Published: June 2021

The vertebrate face is the product of growth and differentiation of structures arising from the pharyngeal apparatus. The arches, pouches, grooves, and membranes that comprise the pharyngeal apparatus undergo morphogenetic movements allowing for a typically seamless integration of tissues and cell types that orchestrate craniofacial development. The differentiation of these structures is critical for facial and skull bones, muscle, connective tissue, skin, and the central and peripheral nervous systems of the head, face, and neck to develop correctly. Hence, it is not surprising that without the proper integration of these tissues during development that craniofacial defects arise, accounting for one‐third of all congenital malformations within human populations. Of central importance, is a thorough understanding of the specific tissue interactions, movements, and fusions that occur between the craniofacial primordia in regulating craniofacial morphogenesis. In this master class, we will discuss the embryology of the head and neck, taking a holistic approach to the components of the pharyngeal apparatus, to review the various elements that come together to form the vertebrate face.Support or Funding InformationNone.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hepatocyte specific ablation of β‐ and γ‐catenin contributes to loss of hepatocyte polarity and EMT induction by activation of the TGFβ pathway
Authors: Pradhan‐Sundd Tirthadipa , Singh Sucha , Molina Laura , Podder Minakshi , Bell Aaron , Monga Satdarshan Pal 
Published: June 2021

A hallmark of epithelial cells is polarization. Like any other epithelial cell, hepatocytes in the liver show a unique polarity by forming several apical and basolateral poles within a cell. Polarization of hepatocytes is necessary for many functions and requires carefully orchestrated cooperation between cell adhesion molecules, extracellular matrix, cell junctions, cytoskeleton, and intracellular trafficking machinery. Loss of hepatocyte polarity is associated with various liver diseases including cholestasis. Previously, we discovered that simultaneous deletion of adherens junction protein β and γ catenin in the mice liver led to cholestasis in the double knockouts (DKO). However, the molecular mechanisms that led to cholestasis in DKO were only partially elucidated. Here, we characterize the phenotype further. Electron microscopy revealed dilation of bile canaliculi with significant reduction in microvilli in the DKO livers. IHC, protein and RT‐PCR analysis revealed loss of several hepatocyte polarity markers including CD10, ZO‐2, BSEP, and MRP2, increased fibrosis and epithelial‐mesenchymal transition in the liver. Remarkably, RNA‐seq analysis revealed over activation of TGFβ pathway in double knock out liver and blocking TGFβ pathway led to partial rescue of tight‐junctional proteins and cholestasis. Taken together, our findings suggest that β; γ‐catenin are critical to the maintenance of hepatocyte polarity and cholestasis development by regulating TGFβ signaling in hepatocytes. Profound understanding of the regulation of hepatocyte polarity by both β ‐and γ‐catenin will be useful not only in understanding cholestasis but also in the context of liver development and pathophysiology.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Quality Control During 40S Ribosome Assembly
Authors: Karbstein Katrin 
Published: June 2021

While much is known about how ribosomes faithfully translate mRNAs into protein, how ribosomes are assembled within cells remains poorly understood. With ~ 2,000 ribosomes synthesized every minute ribosome assembly is the major metabolic activity of a growing cell. Given its profound energy requirements, ribosome assembly is tightly regulated in response to extracellular stimuli. In addition to providing sufficient numbers of ribosomes, cells must also ensure that the produced ribosomes are correctly assembled. This is important to safeguard their catalytic activity and extraordinary fidelity. We are exploring cellular mechanisms to ensure proper assembly of nascent 40S subunits, as well as the consequences of defects in assembly.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Proteoform Dynamics in Steady‐State
Authors: Zecha Jana , Meng Chen , Kuster Bernhard 
Published: June 2021

Utilizing a classical dynamic SILAC (stable isotope labeling of amino acids in cell culture) approach, mass spectrometry‐based technologies nowadays allow for the determination of native protein turnover on a proteome‐wide scale. Obtaining proteoform resolution, however, is often still challenging due to incomplete quantitative information of peptides across pulse time‐points. To address this shortcoming, we evaluated the merits of (a) combining dynamic SILAC with TMT (tandem mass tag)‐labeling of ten pulse time‐points to compute absolute peptide turnover rates, and (b) utilizing solely single time‐point pulses to obtain relative differences in peptide half‐lives.In total, four cell culture replicates of each of the two different methodologies provided turnover information for 80,000 endogenously modified peptides assigned to up to 7,700 protein groups. Protein degradation rates were comparable to those reported with the standard pulsed SILAC approach. Moreover, replicate analyses established that the same reproducibility of turnover determination can be obtained for peptides and proteins eventually facilitating the investigation of proteoform resolved regulation of protein stability. Our comprehensive dataset showed that splice variants can exhibit vastly different cellular protein stabilities. Moreover, it revealed changes in turnover upon post‐translational modifications such as protein phosphorylation, ubiquitination and acetylation on a global scale. Several examples of differentially turned over peptides and proteins were investigated further. For example, differential peptide turnover indicated a previously unknown mechanism of activity regulation via post‐translational destabilization of only the light chain of cathepsin D as well as a phosphorylated version of DNA helicase BLM. Further, a parallel reaction monitoring assay applied to rotenone treated cells suggested that the high protein turnover of members of the respiratory chain complex I is affected by oxidative stress.Our data indicates that the original pulsed SILAC concept can be either expanded by employing TMT labeling of pulse time‐points to estimate absolute peptide turnover rates or simplified by using single time‐point pulses to evaluate relative differences in peptide half‐lives. Both strategies facilitated proteoform resolved turnover measurements. Collectively, they revealed a widely disregarded level of post‐transcriptionally and post‐translationally regulated protein stability which can have important implications for the study of disease mechanisms as well as the assessment of pharmacological interventions.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evidence that Tip60 Induces the DDR & Cardiomyocyte Replicative Senescence in the Neonatal Heart
Authors: Lough John , Lupton Carri , Wang Xinrui , Wan Tina , Auchampach John 
Published: June 2021

Tip60 (Tat interactive protein, 60 kD) is a tumor suppressor protein that acetylates histone and non‐histone proteins. Among the latter, ATM (Ataxia Telangiectasia Mutated), a kinase that induces the DNA damage response (DDR), is activated in cancer cells by Tip60‐mediated acetylation. We are investigating the function of Tip60 in the heart, using global and conditional knockout mouse models to disrupt the Kat5 gene encoding Tip60. Because global knockout (i.e. Tip60−/−) causes early embryonic lethality, we used Tip60+/− heterozygotes to show that, when under cardiac stress, modest Tip60 depletion correlates with re‐activation of the cardiomyocyte (CM) cell‐cycle in the adult heart. More recently, using a knockout model wherein Tip60 is constitutively and specifically depleted in CMs, we observed CM dropout and lethality in three‐month‐old mice, wherein hearts exhibited features of mitotic catastrophe. Because the latter phenotype was preceded by increased CM density, it was suggested that Tip60‐depletion may extend the duration of CM proliferation during neonatal stages of heart development. Moreover, considered in the context of recent reports that ATM is activated at birth to induce the DDR, which in turn causes replicative senescence of CMs by mid‐neonatal stages, these findings suggest that Tip60 induces these events. To examine these possibilities we are conditionally depleting Tip60 in CMs of Tip60F/F;Cre neonatal mice by injecting tamoxifen at postnatal day 0 (P0). To date, immunostaining of sections from neonatal hearts with antibodies that recognize 5′‐BrdU, phospho‐histone‐H3, and phospho‐ATM (pATM) has revealed that Tip60 depletion significantly increases CM proliferation as early as P7, by which time CM proliferation is normally diminished, and that this is accompanied by significantly decreased pATM levels, indicative of a diminished DDR. These findings provide further support for the notion that DDR activation at the time of birth induces replicative senescence of neonatal CMs, implicating Tip60 as the inducer of this process.Support or Funding InformationNIH HL131788 (JL & JA), and a grant from the MCW Cardiovascular CenterThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elucidating the cell signaling pathways for epithelial restitution of a novel probiotic
Authors: Owens Joshua , Saeedi Bejan , Naudin Crystal , Darby Trevor , Jones Rheinallt 
Published: June 2021

Probiotics are a group of beneficial bacteria that when consumed in adequate amounts confer a health benefit. Recently, our group showed that a novel probiotic, Lactococcus lactis sp cremoris (LLC) can elicit strong intestinal cytoprotection in murine experimental models of intestinal damage, and potently enhance restitution of a damaged gut epithelium. The objective of this study was to elucidate the mechanism whereby LLC promotes epithelial restitution. Our goal was to first identify which host pattern recognition receptor that could recognize LLC. We gavaged LLC or vehicle control to mice and assessed transcript enrichment in the colon after 4 hours by RNA‐seq. We detected the specific upregulation of TLR2. We next investigated whether a soluble factor from LLC was able to be detected by TLR2. To test this, we cultured epithelial monolayers with supernatant from an LLC overnight culture and detected elevated TLR2 expression. Furthermore, we detected potent activation of the NF‐kB pathway, and the Nrf2 signaling pathway in epithelial cells following contact by LLC supernatant. Importantly, no negative physiological influences, or cytotoxicity were detected in these cells. Functionally, we found that LLC supernatant accelerated the closure of scratch wound inflicted on an epithelial monolayer in a TLR2‐dpendant manner. These and other data were evidence for us to conclude that LLC mediates epithelial restitution through a TLR2‐dependent pathway. We also propose that cross‐talk between the NF‐kB and Nrf2 cell signaling pathways function in the beneficial effect of LLC on intestinal health and restitution following disease or injury.Support or Funding InformationThis work was supported by the NIDDK‐ R01DK098391This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.