bluuebunny/crossref_metadata_2025_split huggingface dataset:
Safety and Efficacy Study of N‐SORB® a Proprietary KD120 MEC Metabolically‐ Activated Enzyme Formulation: A Randomized, Double‐blind, placebo Controlled Study
Authors: Nair Sreejayan , Guo Rui , Smith Derek , Bisha Bledar , Nair Rama , Downs Berrnard W , Kushner Steve W , Bagchi Manashi 
Published: June 2021

BackgroundEnzymes are crucial for all aspects of metabolic function. Digestive enzymes from natural sources have been credited with beneficial effects in the digestion and absorption of food. N‐SORB is a novel KD120 multienzyme complex (MEC) of metabolically activated enzymes composed of proteases, amylases, lipases, alpha galactosidase, and glucoamylase from natural sources. These enzymes are encapsulated in an SK713 SLP (non‐GMO soy lecithin phospholipid) absorption technology (‘Prodosome®’).ObjectiveThis randomized double‐blind placebo‐controlled investigation assessed the safety and efficacy of Prodosomed N‐Sorb KD120 MEC on various parameters of the blood, immunity, body composition, physical health, and quality of life in young healthy male and female subjects over a 90‐day period.MethodsInstitutional Review Board Approval was obtained from University of Wyoming School of Pharmacy, Laramie, WY. Forty‐six young male and female (mean age: 25.8 ± 12.1 years) healthy volunteers, were randomly assigned to receive either N‐SORB (1 mL, twice daily) or placebo over a period of 90 consecutive days. Complete blood count as well as blood glucose, liver enzymes, and lipid profile were assessed pre‐ and post‐intervention. Serum cytokine levels were determined by using a Bio‐Plex Pro Human Cytokine 8‐plex Assay. Whole body composition analysis was performed by dual‐energy X‐Ray absorptivity (DEXA) to determine body fat mass, lean mass and android and gynoid fat. Body, weight, blood pressure, and physical health were assessed. WHOQOL‐BREF and a Pittsburgh Sleep Quality Index (PSQI) questionnaires were used for quality of life and sleep quality assessments respectively. Adverse events were monitored before, during and after the completion of the study.ResultsOf the 46 subjects enrolled, 41 subjects successfully completed the trial. Compared to placebo, changes in blood cell counts including hematocrit, hemoglobin, mean corpuscular volume, platelets and lymphocytes provide evidence of consistent improvement, which were reconfirmed by Quality of Life (QOL) parameters that significantly improved in the N‐SORB group. No significant changes in BUN, serum creatinine, ALP, AST, ALT, and lipid profile were observed between the placebo and treatment groups. No adverse reports were reported.ConclusionThis randomized, double blind, placebo‐controlled clinical study demonstrates that short‐term intervention with N‐SORB improves the QOL in young healthy volunteers and didn't alter cardiometabolic parameters, lipid profile or body composition. Subjects receiving N‐SORB reported an overall improvement in quality of life.Support or Funding InformationThe Study was Partially Supported by VNI Inc, Lederach, FLThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Optimization of Cell Free Protein Synthesis: Development of a Fusion Protein Fitness Strategy
Authors: Travieso Tatianna , Willis Caroline , Rojas Maria , Eckdahl Anthony , Roland Kyla , Edman Elise , Tovar Sofia Aguilera , Hanlan Nicklaus , Ong Shi Qing , Wagner Jacob , Poet Jeffrey L. , Eckdahl Todd T. , Heyer Laurie J. , Campbell A. Malcolm 
Published: June 2021

Our undergraduate researchers work in the field of metabolic engineering to harness metabolic pathways for the production of useful metabolites with applications in the biofuel, pharmaceutical, or industrial chemical industries. We published a metabolic engineering approach called programmed evolution that introduces variation in regulatory elements for the expression of enzymes that control orthogonal metabolic pathways. Programmed evolution employs a riboswitch‐based fitness module to transduce the production of a desired metabolite into antibiotic resistance. We validated programmed evolution by optimizing the conversion of caffeine to theophylline by caffeine demethylase, but we recognized several limitations to the widespread use of our approach, including crosstalk between orthogonal and native bacterial metabolism, potential toxicity of metabolites, and poor representation of large variation spaces. To avoid these limitations, we are developing an in vitro approach to programmed evolution. For cell‐free protein synthesis (CFPS), we found supercoiled plasmid was a much better template than linear PCR amplicon. We designed, constructed, and have begun testing a novel fitness module that responds to the production of theophylline produced by the action of caffeine demethylase produced by CFPS. The fitness module encodes a fusion protein composed of three functional units. The amino portion contains a Gal4 DNA binding domain that will target binding sites on a library of DNA regulatory elements for the expression of caffeine demethylase. The carboxyl terminus includes streptavidin which will bind to avidin beads for physical separation of selected DNA regulatory elements. Sandwiched in between these two domains is GFP, which serves as a biosensor. Using this in vitro selection strategy, we plan to pursue the optimization of CFPS for the discovery of efficient metabolic pathways of interest.Support or Funding InformationNSF RUI grant MCB‐1613203 to Davidson College and MCB‐1613281 to Missouri Western State University, Davidson College Martin Genomics ProgramThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antimicrobial Activity of 3, 4‐Annelated Coumarin‐Benzodiazepine‐Systems
Authors: Govori Sevdije , Ibrahimi Hamide , Daci Majlinda , Haziri Arben 
Published: June 2021

This study presents the synthesis of 7‐iminobenzo[b]chromeno[4,3‐e][1,4]diazepin‐6(7H)‐ones (3–4) by simple reaction of 4‐chloro‐2‐oxo‐2H‐chromene‐3‐carbonitrile with o‐phenylidenediamine. The compounds (3–4) were characterized by elemental analysis data, IR, 1H NMR and mass spectral data. The novel heterocyclic compounds were evaluated for antimicrobial activity against Escherichia coli and Staphylococus aureus using the Kirby‐Bayer method. The novel compounds demonstrated moderate antimicrobial activity.Support or Funding InformationFunding: Support for this study was provided by the Ministry of Education, Science and Technology of the Republic of Kosovo. Grant number:2‐5521‐2This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Alcohol + PLD = Phosphatidylethanol, a Long‐Term Alcohol Biomarker
Authors: Neilsen Paul O. , Johnson L. Jeff , Rzepecki Piotr , Davies Sean 
Published: June 2021

Heavy alcohol consumption—both chronic and binge, increases the risk of digestive tract tumors, coronary artery disease, and other health problems. Better biomarkers for consumption and ethanol‐induced organ damage are needed. Direct biomarkers including phosphatidylethanol (PEth), ethyl glucuronide (EtG) and ethylsulfate (EtS) are gaining increased use and acceptance. We first prepared bioanalytical tools and assays then recruited 9 alcohol‐consuming volunteers (2 Heavy, 4 Moderate, 3 Light) in a pilot study. We measured four abundant PEth species by mass spectrometry and anti‐PEth IgG antibodies by ELISA. We found a high correlation between total PEth measured and expected PEth levels depending on amount and timing of ethanol consumption. Interestingly, we observed differences in PEth molecular species based on the amount of alcohol consumed, and even differences between the two heaviest drinkers—one of which drank in a more regular pattern while the other had a number of binge episodes. These observations led to a clinical study where alcohol was administered in both binge and consistent patterns with abstinence periods monitored by continuous alcohol monitoring (CAM) ankle bracelets. We sampled urine for EtG/EtS; serum/plasma for both carbohydrate deficient transferrin (%CDT) and anti‐PEth IgG; and whole blood for PEth. We found the direct biomarkers (PEth, EtG/EtS) were superior to indirect biomarkers (%CDT and anti‐PEth IgG) for detecting alcohol consumption for both drinking patterns at 1, 7 and 21 days post‐consumption. Interestingly, we found that plasma anti‐PEth IgG levels followed blood alcohol concentration in binge‐drinking volunteers indicating preformed, bound IgG is displaced by high blood alcohol levels. Our results indicate that PEth is a valuable biomarker for detecting both acute and consistent alcohol consumption; and that this unique lipid merits further study to better understand its formation, elimination, and impact on the human immune system and alcohol‐related disorders.Support or Funding InformationSupported by National Institutes of Health, NIAAA, Contract, HHSN267200800020C to PON.The content of this communication is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute on Alcohol Abuse and Alcoholism nor the National Institutes of Health.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fluorogenic Atom Transfer Radical Polymerization as a Strategy for Biomolecular Detection
Authors: Tahseen Danyal , Cooley Christina B 
Published: June 2021

The sensitive detection of analytes is valuable in various chemical and biomedical contexts. This sensitivity can be conferred by signal amplification, where the readout magnifies the signal of an otherwise low level of analyte. The Cooley Lab's approach to signal amplification involves using fluorogenic radical polymerizations, where the polymers' fluorescence is a quantifiable proxy for presence of an analyte. This approach also enables the qualitative appreciation of analyte presence since the fluorescence is visible to the naked eye.In order to use this technology to detect water‐soluble analytes such as biomolecules, we are conjugating the polymerization initiator to a model protein. This involves developing the fluorogenic polymerization in biologically relevant conditions such as an aqueous medium, so that polymers can be grafted directly from the biomolecule. The choice of an atom transfer radical polymerization (ATRP) facilitates the controllable formation of fluorogenic polymers in aqueous media as a function of both analyte concentration and reaction time. The development of the polymerization towards a biomolecular detection assay will be discussed, with emphasis on optimization of the reaction conditions, synthesis and characterization of biomolecule‐initiator conjugates, and fluorescence detection.Support or Funding InformationSan Antonio Area Foundation, American Chemical Society Petroleum Research Fund, Trinity UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A New Synthetic Route to Cycloheptatrienylidene Fluorophores
Authors: David Navindra , Hyatt Ivan Fabe Dempsey 
Published: June 2021

Luminescent molecules can lose their fluorescent properties through aggregation caused quenching (ACQ). ACQ is caused when these molecules, in high concentration, undergo π‐π stacking. Previous methods have been developed that utilize Aggregate Induced Emission (AIE) mechanisms to avoid the π‐π stacking. Analyzing how to turn on this AIE mechanism through aggregation with a metal can stop this stacking phenomena and allow the molecule to exhibit fluorescence in a concentrated solution. One known fluorophore that exhibits AIE properties are cycloheptatrienylidenes (CHT). The project described herein uses hypervalent iodine in a new methodology to synthesize CHTs. A faster and cheaper way to synthesize asymmetric CHTs is being investigated for the generation of AIE materials. Currently, methodologies of producing symmetrical cycloheptatrienylidene fluorophores are expensive, time consuming, and involve multiple steps. Ongoing reactions involve synthesizing the CHT using a modified Catellani reaction. The Catellani reaction has eliminated a number of steps in creating the precursor to the CHT. Production of AIE fluorophores that will “turn‐on” fluorescence after coming into contact with metals could be used to activate endogenous metals in the body which could help to target malignancies such as cancer.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 Computational Evaluation of Biological Immunofluorescence Data
Authors: Hietpas Taylor 
Published: June 2021

Optical spectroscopy and fluorescence have become important techniques for elucidating molecular biological function. Two‐photon and confocal microscopy have particularly enabled increased potential for data acquisition and cellular resolution. With this increased potential has developed a need for efficient and reliable methods for evaluating these large data sets. Historically, researchers have relied upon fundamental image processing techniques and manually implemented image modification to obtain interpretations of their data. While these methods have proven successful, increased computation power and well‐established statistical segmentation protocols have enabled a renaissance of sorts in biological image analysis. In our lab, lymphatic and macrophage immunofluorescence data or endogenous fluorescence from transgenic reporters is most commonly acquired via either confocal or two‐photon microscopy of whole mount corneal and skin tissues. Manual analysis of these large and complex data sets is time consuming and technically challenging. This project has allowed us to explore more sophisticated methods of image processing and analysis. First, K‐means clustering algorithms were implemented to evaluate efficacy of unsupervised partitioning and segmentation of these image data sets. While K‐Means has proven effective at segmentation in many disciplines, the task of segmenting noise and autofluorescence has in these types of data has proven subtler than expected. As such, further methodologies are being investigated including Nonnegative Matrix Factorization, Spectral Phasor Analysis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activating transcription factor 4‐dependent B cell translocation gene‐1 induction promotes ferroptosis
Authors: Cho Il Je , Kim Jae Kwang , Kim Eun Ok , Park Sang Mi , Ki Sung Hwan , Kim Sang Chan 
Published: June 2021

Ferroptosis is a special type of programmed cell death that is characterized by iron‐dependent lipid peroxidation. The present study investigated the role of B cell translocation gene‐1 (BTG‐1) on sulfur amino acid deficiency (SAAD)‐mediated cell death. SAAD in HepG2 cells depleted the reduced glutathione, accelerated lipid peroxidation, increased cyclooxygenase 2 expression, and promoted cell death. SAAD‐mediated cell death was specifically inhibited by pretreatment with ferrostain or defroxamine, chemical inhibitors of ferroptosis. In parallel with cell death, SAAD increased expression of microtubule‐associated protein light chain 3‐II and facilitated acridine orange positive autophagic flux. Inhibition of autophagy accelerated the SAAD‐mediated cell death. Moreover, SAAD induced BTG‐1 expression in HepG2 cells. By using BTG‐1 promoter haboring reporter gene and siRNA, activating transcription factor 4 (ATF4) was identified as an essential transcription factor for SAAD‐mediated BTG‐1 induction. Knock out of BTG‐1 gene by CRISPR/Cas9 system decreased the cytotoxicity by SAAD. Furthermore, BTG‐1 deficiency increased microtubule‐associated protein light chain 3‐II and autophagic flux. Thus, present study demonstrates that SAAD induces ferroptosis, and the ATF4‐dependent BTG‐1 induction contributes to accelerate ferroptosis by inhibiting autophagy.Support or Funding InformationThis study was supported by the National Research Foundation of Korea (NRF) funded by Korea government (MSIP)(Grant No. 2018R1A5A2025272 and 2018R1A2B6007514).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Chloroform Fraction of Adenopus Breviflorus Benth Fruit on Opening of Rat Liver Mitochondrial Permeability Transition Pore, Mitochondrial Atpase and Cytochrome C Release
Authors: Oyedeji Tolulope A , Akobi Chibuzor I , Onireti Daniel O , Olorunsogo Olufunso O 
Published: June 2021

Bioactive agents from medicinal plants that can induce the intrinsic pathway of apoptosis in tumours by opening of the mitochondrial permeability transition (MPT) pore maybe developed into drugs useful in chemotherapy. The fruit of Adenopus breviflorus (A. breviflorus), a tropical medicinal plant found in the Western part of Nigeria, West Africa, is widely used in folklore medicine against the outbreak of measles, and in the treatment of gonorrhoea in men. However, its use in the treatment of tumours has not been well elucidated. Therefore, this study aim at assessing the in vitro effect of varying concentration of the chloroform fraction of A. breviflorus (CFAB) on the opening of rat liver MPT pore, mitochondrial ATPase (mATPase) and cytochrome C (Cyt C) release. Rat liver mitochondria were isolated by differential centrifugation. Mitochondrial Permeability Transition, mATPase activity and Cyt C release were determined spectrophotometrically. The results obtained from this study show that the varying concentration of CFAB, significantly induced opening of MPT pore in rat liver mitochondria in a concentration‐dependent manner. Specifically, the pore was opened by 15.0, 18.8. 18.9 and 19.2 folds at 50, 150, 250 and 350 μg/mL, respectively. Also, the CFAB also stimulated mATPase activity by 4.4, 2.5, 3.3 and 22.4% at 50, 150, 250 and 350 μg/mL, respectively. Furthermore, the varying concentration of CFAB significantly (p < 0.05) stimulated Cyt C release in concentration‐dependent manner. This study therefore suggests that the chloroform fraction of A. breviflorus may possess potent bioactive agents that could induce the intrinsic pathway of apoptosis through the induction of the mitochondrial permeability transition pore, stimulation of mitochondrial ATPase and the release of cytochrome c.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Epigenetic Modifiers on PD‐L1 and HLA Class I Expression on Tumor Cells
Authors: Maini Trisha , Sternberg Lauren , O'Donnell Robert W. 
Published: June 2021

Expression of PD‐L1 on various tumor cell lines leads to the inhibition of T cell cytotoxicity. When this takes place, the T cells are unable to combat the tumor cells and cancer persists. In the same regard, increased expression of human leukocyte antigen (HLA) allows tumor cells to be more easily detected by these T cells. In previous research, epigenetic modifiers including the DNA methylation inhibitor 5‐Azacytidine (5‐AzaC) and the histone deacetylase inhibitor vorinostat, were shown to increase HLA expression. However, epigenetic modifiers have also been shown by others to upregulate PD‐L1 expression. We hope to elucidate whether or not PD‐L1 expression is upregulated along with HLA expression upon exposure to these epigenetic modifiers. If this were to occur, then the effects of both PD‐L1 and HLA would essentially negate each other. Two cell lines that are known to express PD‐L1 are MCF‐7 and MDA‐MB‐231. Our initial results confirm that PD‐L1 is expressed on the MDA‐MB‐231 cell lines, as demonstrated by flow cytometry. Our results have also shown that two other cell lines, MDA‐MB‐435 and HL‐60, do not express PD‐L1 and that exposure with the epigenetic modifier 5‐AzaC did not cause either cell line to express PD‐L1. Nonetheless, there was still an up regulation of MHC Class I antigens following a 48 hour exposure to a non‐cytotoxic dose of 5‐AzaC. Future experiments are planned to optimize the doses of 5‐AzaC and vorinostat to maximize HLA expression while not increasing or inducing the expression of PD‐L1.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of Sodium Glucose Transporter 2 Inhibitors on Proliferation and Growth Factor Signaling Pathways in Triple Negative Breast Cancer
Authors: Ware Kierra , Smith Tunde , Brown Del‐Vecie , Hill Dorian , Stewart LaMonica 
Published: June 2021

Triple negative breast cancer (TNBC) is an aggressive form of breast cancer that does not express the estrogen receptor, progesterone receptor, and the HER2 receptor. Since it does not express these receptors, TNBC does not respond to many of the standard therapies for breast cancer. To decrease the number of deaths associated with breast cancer, we must identify therapeutic strategies that effectively prevent the growth and progression of TNBC. Recent studies from our laboratory and others show that the sodium glucose transporter 2 (SGLT2) inhibitor canagliflozin reduces proliferation of human prostate cancer cell lines. To determine whether SGLT2 inhibitors can also suppress TNBC growth, we tested the effect of canagliflozin and other SGLT2 inhibitors on proliferation and protein expression within the BT‐549 human breast cancer cells. Presto Blue assays revealed that two SGLT2 inhibitors, canagliflozin and ipragliflozin, significantly inhibited BT‐549 cell proliferation. However, the SGLT2 inhibitor empagliflozin produced little to no change in cell proliferation. We next examined the effect of SGLT2 inhibitors on Akt and Erk ½ MAP kinase, two proteins that promote TNBC growth. Western blot analysis revealed that canagliflozin reduced phosphorylation of Akt and Erk ½ within BT‐549 cells. Therefore, the ability of canagliflozin to block BT‐549 proliferation may be due in part to canagliflozin‐mediated decreases in Akt and Erk activity. The epidermal growth factor receptor (EGFR) activates both Akt and Erk ½ signaling in BT‐549 cells. To determine whether inhibition of EGFR signaling would alter the response to canagliflozin, we tested the combined effect of canagliflozin and EGFR inhibitors on BT‐549 cell proliferation. Canagliflozin alone was more effective at reducing BT‐549 cell proliferation than the EGFR inhibitors gefitinib and lapatinib. However, the combined effect of canagliflozin and gefitinib as well as the combined effect of canagliflozin and lapatinib was greater than that produced by either compound alone. These data suggest that combination treatments involving canagliflozin and EGFR inhibitors may serve as an effective therapy for patients with TNBC.Support or Funding InformationNIH grant 5T34GM007663This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.