bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cryogenic Grinding and 3D Printing Techniques for Establishing “Disperse and Absorb” Brick‐Type Constructs of Food Materials
Authors: Rhee Jin‐Kyu 
Published: June 2021

The potentialities of 3D printing technology are discussed from technical and research‐oriented perspectives for industrial manufacturing of a variety of food products. Currently, 3D printing technology has been advanced to enable us to process or cook innovative foods. However, food‐based materials for 3D printing are still limited in terms of eating qualities, nutritional values and functionality as well as industrial production. Therefore, this uprising issue on alternative food processing techniques especially focused on the exploration of new food materials combined with these 3D printing technologies needs to be re‐spotlighted, and then solved to pave the way to this innovative and sensational area of investigation with more accessibility. In this work, fabrication parameters were characterized and optimized to improve printability, quality and efficiency of manufacturing foods. First, carbohydrates/protein‐based food materials were cryo‐powdered to render an amorphous shaped‐ micronized‐ materials with characterization. Subsequently, modification of structure with this micro‐sized food materials has been re‐crystallized by controlling water contents and heat. Then, bonding materials has been formulated and jetted onto the carbohydrates/protein‐based food powders to be polymerized. Construction of cohesive micro‐sized geometry through combining materials with controllable parameters was optimized by the responsive surface methodology. This technology has a potential to design the exterior of foods and internal microstructure to give texture with controlled body‐absorption.Support or Funding InformationThis research was supported in part by the High Value‐added Food Technology Development Program, the Ministry of Agriculture, Food and Rural Affairs (MAFRA), Republic of Korea (316058‐3, 315063‐3), the Functional Districts of the Science Belt support program, Ministry of Science, ICT and Future Planning, Republic of Korea (2017K000017), and the Ewha Womans University Research Grant of 2017.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Considerations for Western Blot Normalization Techniques
Authors: Oh Kenneth , Dreskin Elizabeth Jordan , Liu Ning , Hammond Matthew , Kollmann Franziska , Posch Anton 
Published: June 2021

Western blotting is a well‐established method to specifically measure the relative quantities of individual proteins in complex biological samples. However, data quality may be affected by technical and biological variation. The application of data normalization strategies is strongly recommended to compensate for these non‐sample related variations in signal intensity. Here, we compare the power of two different data normalization strategies, namely housekeeping protein (HKP) and total protein (TP) normalization based on Stain‐free technology to offer an orientation in selecting the appropriate data normalization tool. In brief, HeLa cell lysates were separated with SDS‐PAGE Stain‐free gels and blotted onto a PVDF membrane. Stain‐free imaging after activation with UV was applied to obtain a total protein blot signal. The blot was then treated to the appropriate immuno‐probing steps and the intensity of the target protein ATG7 was recorded by chemiluminescence. In order to illustrate the differences of the two techniques, the membrane was subsequently immuno‐probed for GAPDH as a typical example for housekeeping protein data normalization. Both data sets were used for data normalization of the target protein ATG7. Our results revealed that total protein staining through Stain‐free technology shows less technical variation than HKP recording as represented by GADPH. Additionally, the use of housekeeping proteins as data normalization tool is restricted in its general applicability and may not be compatible with the strict publishing journal guidelines.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An Aptamer‐Based Approach to Assess the Human Plasma Proteome for Pre‐Analytical Variability
Authors: Daniels Jaclyn R. , Cao Zhijun , Maisha Mackean , Schnackenberg Laura K. , Sun Jinchun , Pence Lisa , Schmitt Thomas C. , Kamlage Beate , Beger Richard D. , Yu Li‐Rong 
Published: June 2021

The quality of clinical samples is of utmost importance when conducting biomedical research with OMICS‐based assays as the variability introduced before sample analysis is a potential confounder in the results. In this study, we used the aptamer‐based SOMAscan® proteomic technology to characterize pre‐analytical variability in human EDTA plasma. The goal of the study was to investigate how variation in the pre‐analytical steps during processing of whole blood to plasma impacts the plasma proteome. Whole blood was obtained from apparently healthy individuals and was collected in a randomized sequence of six EDTA tubes. The pre‐analytical variables included prolongation of the processing start time, storage temperature of whole blood or plasmaprior to and after processing, and variation in applied centrifugal force. The SOMAscan® assay was performed to measure the relative abundance of 1305 proteins. To determine the effects of variations inpre‐analytical processing, the data was statistically analyzed and the results were used to identify significant changes in protein abundance under each study condition relative to the control condition (samples processed without delay and immediately stored at −80°C). Both low centrifugation force(1300 × g compared to 2500 × g) and blood storage at 0°C for 6 hours resulted in the most significant changes in protein abundance (200 and 148 proteins, respectively, fold change ≥ 1.2, P < 0.05, and FDR < 0.05), while storing immediately processed plasma for 24 hours at 4°C yielded the least changes (28 proteins). The results indicate that blood samples should be immediately processed to plasma and frozen in −80°C without delay in order to preserve sample integrity and to minimize any proteome abundance alterations. The results also provide important information on the discovery of protein abundance changes due to pre‐analytical variability that can be useful in performing effective quality control assessments.Support or Funding InformationThis study was supported with funds from the National Center for Toxicological Research, Food & Drug Administration, Jefferson, Arkansas.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bacterial Viruses Organize Subcellular Environments to Mediate Heterogeneous Development
Authors: Trinh Jimmy Tri , Zeng Lanying 
Published: June 2021

Spatial organization underpins biological processes in lifeforms. For complex organisms, body parts must develop properly in space, and within the cells making up these organisms, membranes separate organelles with different functions. Simpler organisms, like bacteria, lack intracellular membranes, but possess alternate strategies for organization, where certain proteins are confined to specific cellular locations to consolidate their functions. Viruses, among the simplest lifeforms, have also been reported to display organized development, further underscoring the essentiality of structure in biology.Bacterial viruses, known as bacteriophages or phages, are utilized as model systems for studying advanced cellular functions. Phage lambda infects Escherichia coli and propagates via distinct pathways, either lysis or lysogeny, where this fate‐selection is a paradigm for cellular decision‐making. Recent efforts probing decision‐making at the subcellular level have discovered that phages behave as individual interacting intracellular entities when processing decisions, generating considerable insights into how complex decision‐making may occur. However, little is known about this phage's spatial development within its host and how spatial organization might affect cellular decision‐making.Here, we characterize how phage lambda develops in subcellular space, from infection through lysis, using multiple fluorescence reporters targeting phage DNA replication, essential host resources, phage transcription, and phage assembly. We observe that these processes coalesce into distinct areas of the host cell during infection to coordinate phage development, reminiscent of virus factories, where multiple factories may be present in single cells. Remarkably, different intracellular factories quantitatively diverge at the viral DNA, mRNA and protein levels, suggesting the existence of bona‐fide microenvironments within cells. We propose that individual viruses drive the formation of disparate territories, implying that phages may sense separate environments during decision‐making and development. The conclusions from this work may shed significant light on the detailed molecular mechanisms of cell‐fate bifurcation.Support or Funding InformationThis work was supported by the National Institutes of Health (R01GM107597).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RXRA is a direct target gene of miR‐506‐3p that regulates oncogene MYCN expression and cell differentiation in neuroblastoma
Authors: Shelton Spencer Dewayne , Zhao Zhenze , Du Liqin 
Published: June 2021

Neuroblastoma is an aggressive childhood cancer that arises from neural precursor cells that fail to differentiate. In 20% of high‐risk neuroblastoma cases oncogene MYCN is overexpressed. We previously identified that miR‐506‐3p is a strong repressor of MYCN expression as well as an inducer of neuroblastoma differentiation. However, the 3'UTR of MYCN mRNA lacks the target site of miR‐506‐3p, suggesting that miR‐506‐3p regulates MYCN expression through an indirect pathway. Combining gene expression array data and informatics analyses, we identified 11 candidate genes that are direct targets of miR‐506‐3p and potentially regulate MYCN expression. The candidate genes were silenced and screened for decreased MYCN expression and increased neuroblastoma neurite outgrowth – the morphological differentiation marker of neuroblastoma cells. Interestingly, retinoid X receptor alpha (RXRA) was the only gene to exhibit significant downregulation of MYCN expression and induced the most dramatic neurite outgrowth. We then examined the RXRA protein expression during retinoic acid treatment and demonstrated that RXRA protein level is significantly downregulated in retinoic acid treated cells. Upon investigation of neuroblastoma patient databases, we found that high risk and MYCN amplified patients that exhibited higher RXRA mRNA levels had lower overall and relapse free survival statistics. Altogether, these results suggest that RXRA functions as an oncogene through promoting MYCN expression and repressing cellular differentiation in neuroblastoma. Further investigation is ongoing to elucidate the molecular pathways that mediate the oncogenic function of RXRA in neuroblastoma cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigation of Domain Responsible for Calcineurin B Homologous Protein (CHP) Isoform Specific Function
Authors: Davis Shane , Wallert Mark , Provost Joseph 
Published: June 2021

Mammalian cells ubiquitously express the Sodium‐Hydrogen Exchanger (NHE1) a membrane transporter responsible for intracellular pHi, motility and proliferation. Calcineurin B Homologous Protein (CHP) regulates activation and kinetics of the NHE1. CHP has two isoforms, CHP1 and CHP2, whose physiological function remains unclear but both have a unique role in nascent tumor survival. There is a signature domain in the sequence of an otherwise homologous structure between CHP1 and CHP2 that we believe is responsible for CHP isoform specific function, including regulation of NHE1. The purpose of this study is to understand how the these domains of CHP1 and CHP2 confer to the properties of the CHP, particularly cell proliferation and migration. To identify possible differences of structural conformation that could be related to function of each CHP isoform we exchanged the (aa‐aa ofr CHP1 and aa‐aa of CHP2 using gibson cloning. Gibson cloning allows us to manipulate fragments created by PCR to make mutants of our wild‐types. These constructs were then transiently transformed into cells and changes in proliferation and migration analyzed compared to wild‐type expressing cells. We hypothesize that there is an interaction between CHP1 and CHP2 that allow for their individual absence or presence to affect the proliferation and motility function in lung carcinoma.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of hydrolytically activated, oxygen‐generating biomaterials to enhance drug efficacy on cultured ovarian cancer cells
Authors: Clark Matthew Paul , Kraus Aileen , Higgins Jillian , Gallant Benjamin , Arruda Dominic , McTigue Christie , Li Xiaofei , Guo Hao , Chen Hongduo , Cui Yali , Di Wen , Wan Yinsheng 
Published: June 2021

Ovarian cancer remains the most prevalent type in gynecological cancer, with drug resistance still being a challenge. Our previous studies have demonstrated that ovarian cancer cells are resilient due to their active EGFR and AKT/mTOR pathways. Also, abundant data suggests that similar to other cancer types, ovarian cancer cells grow and proliferate remarkably well under hypoxia situation. We hypothesize that the application of oxygen to ovarian cancer cells may render unfavorable conditions, thus leading to the enhanced cell death, when combined with chemotherapy drugs. Recent studies have shown that polydimethylsiloxane (PDMS)‐encapsulated calcium peroxide, or PDMS‐CaO2, potently deliver oxygen and eliminate hypoxia‐induced cell dysfunction and cell death in normal pancreatic cells and other types of normal cells. In this study, we followed the published protocol, produced PDMS‐CaO2 disks and tested in vitro the effects of released oxygen on cultured ovarian cancer cells (CaOV3 cells). The results showed that our manufactured disks sustainably release O2 in a time dependent manner. MTT assay results indicated that PDMS‐CaO2 disks induce cell death in an oxygen dependent manner, compared to PDMS disks only. Combination of inhibitors of EGFR, MEK/ERK and AKT/mTOR with PDMS‐CaO2 disks increases cell death. Interestingly, PDMS disks significantly enhance the effects of cisplatin, oxaliplatin and doxorubicin, but not that of paclitaxel or taxol. PDMS‐CaO2 disks inhibit exosome inhibitor GW4869 induced cellular proliferation. Also, those disks significantly augment ionophore monensin A (for Na+/H+ antiporter), and calcium ionophore A23187‐induced cell death. Confocal microscope data showed that PDMS‐CaO2 disks alter mitochondria activities. Collectively, our data suggests that PDMS‐CaO2 disks releasing O2 and inducing cell death may affect cell membrane ion channels and mitochondria activities. The combination of cancer drugs or ionophores or EGFR/MEK/AKT/mTOR inhibitors with PDMS‐O2 disks may provide novel approaches for ovarian cancer clinical management.Support or Funding InformationNIHThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Impacts of Hypoxia on Na+/H+ Exchanger Isoform 1 and Calcineurin B Homologous Protein Isoform 2 Expression in Non‐small Cell Lung Cancer
Authors: Wallert Clarice H. , Provost Joseph J. , Wallert Mark 
Published: June 2021

The Na+‐ H+ exchanger isoform 1 (NHE1) is a key regulator of cell proliferation, migration and invasion in cells from a variety of solid tumors. The calcineurin B homologous proteins (CHP1 and CHP2) appear to be essential cofactors to support NHE1 function. CHP1 appears to be expressed ubiquitously in healthy tissue, while CHP2 is predominantly expressed in tumor cells. While both CHP isoforms are highly homologous and bind in nearly identical regions on NHE1, each has been identified to have a distinct impact on NHE1 function. Our research has demonstrated an elevation in CHP2 expression in tumor samples from patients with either adenocarcinoma or squamous cell carcinoma of the lung as well as in a broad range of NSCLC cell lines. To evaluate the role of CHP2 in NSCLC cells we used NCI‐H1299, a carcinoma cell line, as the parent cell line and developed two new cell lines. One of these cell lines lacked CHP2 expression (H1299 CHP2KD) and the other lacked NHE1 expression (H1299 NHE1KD). We hypothesized that CHP2 expression enhances cell survival and the tumorigenic potential of these cells. We evaluated the impact of a low serum and hypoxic microenvironment on CHP2 expression and determined CHP2 levels increased when H1299 cells are grown in 0.5% serum and 1% O2 compared to cells grown in 10% serum and 21% O2 (normal conditions) after 24 and 48 hours; however, at approximately 72 and 96 hours, CHP2 expression in the same conditions decreased while NHE1 expression increased. CHP2 may be involved in initial survival mechanisms during the time NHE1 expression is increasing to further allow for cancer progression. To evaluate the role of CHP2 and NHE1 biologically, proliferation and migration were assessed in all conditions. Proliferation and migration rates in normal conditions for H1299 CHP2KD cells were approximately 50% of H1299 cells. When these experiments were performed in low serum and hypoxic conditions, the proliferation and migration rates of the H1299 cells was reduced to about 50% of that seen in normal conditions, while the rates of proliferation and migration in H1299 CHP2KD was reduced to a rate equivalent to the H1299 NHE1KD cells. These data suggest a major role for CHP2 expression in the development and progression of non‐small cell lung cancer.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ascites tumor microenvironment and chemoresistance in ovarian cancer
Authors: Kim Soochi 
Published: June 2021

Ovarian cancer is inherently silent; most patients are diagnosed in late stage of the disease progression. The progression and resistance to chemotherapy in ovarian cancer patients is closely associated with aberrant accumulation of fluid in the peritoneal cavity, called ascites formation. Malignant ascites constitutes a unique tumor microenvironment providing a physical structure for the accumulation of both pro‐tumorigenic and anti‐tumorigenic factors, each of which cumulatively favor cancer cell proliferation, metastasis and chemoresistance. Almost all patients with recurrent ovarian cancer are presented with ascites and have acquired resistance to chemotherapy. However, underlying mechanism of ascites induced acquired resistance to chemotherapy have not been fully understood yet. We used patient derived ascites to delineate the role of ascites in acquired chemoresistance in ovarian cancer. Of those many soluble factors, we focused on the levels of cholesterol in malignant ascites. We found that cholesterol levels are highly elevated in malignant ascites to that of benign peritoneal fluid from ovarian cyst patients. Treatment of cholesterol significantly induced resistance to cisplatin and paclitaxel via increased expression of drug resistance proteins, ABCG2 and MDR1, together with upregulation of LXRα/β. Moreover, LXRα/β expression was positively correlated with MDR1 but not with ABCG2 expression in ovarian cancer patient‐derived ascites cells. The importance of cholesterol induced LXRα/β in acquired resistance were further confirmed by silencing LXRα/β. In addition, elevated cholesterol in malignant ascites was associated with shorter recurrence free survival in ovarian cancer patients. Collectively, these results suggest that cholesterol in ovarian cancer patient derived ascites causes resistance to cisplatin through upregulation of MDR1 via activation of LXRα/β in ovarian cancer. Targeting the pathological interaction between cancer cell and ascites tumor microenvironment may be an efficient strategy to overcome acquired resistance in ovarian cancer patients.Support or Funding InformationThis research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HA17C0037, HI16C2037) and grants from Seoul National University College of Medicine (800‐20150304) and the National R&D Program for Cancer Control, Ministry of Health & Welfare, Republic of Korea (1720110).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enhancement Natural Killer Cell Activity of Fucoidan in Lung Metastasis in vivo Model
Authors: Kim Jinhye , Kim Da Hyun , Chun Su‐Hyun , Park Ho‐Young , Lee Kwang‐Won 
Published: June 2021

Many people are interested in well‐being in their lives. Chronic diseases like cancer or hypertension cause low quality of life in those having those diseases. As bioactive substances, fucoidan is helpful to people. Also fucoidan is a polysaccharide with sulfate group and investigated in this study. Fucoidan is well known to be effective in improving the immune system, anti‐virus effect, cardiovascular system, and anti‐cancer effect. The object of this study is to investigate the anti‐tumor effects of fucoidan on lung metastasis in BALB/c mice. Experimental animals were randomly divided into three groups: (1) control group, (2) cancer control group, and (3) group fed fucoidan (60 mg/kg B.W.). Each group was fed orally once per day for 6 weeks. There were no differences of body weights, chow and water intake among three groups. Natural killer (NK) cell activity tested by LDH assay was significantly different (p<0.05) between cancer control group and group fed fucoidan. But, the number of tumor colony and the concentration of NO in serum were not significantantly different among three groups (a control group, a cancer control group, and a group fed fucoidan). We confirmed that fucoidan can improve NK cell activity in lung metastatis in vivo model.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.