bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Time Course Analysis of the Effects of Arachidin 1 and Arachidin 3 on a Rotavirus‐Infected Human Intestinal Cell Line
Authors: Witcher Caleb Michael , Lockwood Hannah Nicole , Napier‐Jameson Rebekah , Mattila Macie N , Strange Essence B , Taylor Josephine , Clack Beatrice , Medina‐Bolivar Fabricio , Ball Judith M 
Published: November 2023

Rotavirus (RV) causes severe gastroenteritis in infants and children worldwide. To prevent RV induced diarrhea, research efforts are focusing on new vaccines and novel therapeutic approaches. Previously, our laboratory has demonstrated that arachidin 1 (A1) and arachidin 3 (A3) have significant effects on the amount of infectious RV particles released from a human intestinal cell line, HT29.f8, at 18 hours post infection (hpi). Morphometric analyses have suggested changes in programed cell death pathways. The objective of this study was to observe the effects of A1 and A3 on RV infected HT29.f8 cells at 12, 14, 16, 18 hpi. A time course morphometric analysis was performed to visualize the effects of the arachidins on cellular ultrastructure and RV particles using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Additionally, changes in ratios of nucleus to cytoplasm during the time course were determined with/without RV and arachidin treatments. RV particles (with/without envelope) in infected cells (with and without the arachidins) were measured and statistically compared to determine if the observed virus particles represented different populations. To quantify the infectious virus particles released from the RV infected cells, plaque forming assays were performed. Additionally, tunable resistive pulse sensing technology (TRPS) using the qNano system from Izon was used to quantify total infectious virus particles and determine the size distribution of the released RV particles from HT29.f8 cells with/without the arachidins during the time points of collection. qRT‐PCR was performed to further identify changes in the expression of key apoptotic and autophagic genes, and western blots assays were used to confirm changes in the expressed proteins that are essential for the two cell death pathways. The data suggests that the arachidins have a significant role in influencing HT29.f8 cells to enter apoptotic or autophagic cell death pathways, and inhibit the maturation and production of progeny RV populations. The plaque assays confirmed a change in the amount of infectious virus particles produced with the addition of thee arachidins. This implies that A1 and A3 arrest RV particles at the enveloped stage of development. Thus, these small molecules have the potential to be developed as anti‐RV therapeutic agents.Support or Funding InformationThis work was supported by: 1) The Office of Research and Sponsored Programs at Stephen F. Austin State University (RPS # 107552‐26112‐150); 2) The NSF‐EPSCoR (grant# EPS‐0701890; Center for Plant Powered Production‐P3).
Peptide B7–33, a Single‐Chain Derivative of the Relaxin Hormone Protects Cytotrophoblasts from Marinobufagenin‐Induced Preeclampsia Phenotype
Authors: Pantho Ahmed Faiaz , Afroze Syeda Humayra , Dean DeShaundra M , Hossain Mohammed Akhter , Bathgate Ross , Xia Sijie , Zawieja David C , Kuehl Thomas J , Uddin Mohammad Nasir 
Published: November 2023

BackgroundPreeclampsia (PreE) is a hypertensive pregnancy disorder, which occurs in approximately 10% of all gestations. Recently, a digitalis‐like factor, marinobufagenin (MBG) has been implicated as a causative factor in preE. We demonstrated that MBG inhibits the proliferation, migration, and invasion of cytotrophoblast (CTB) cells. Relaxin is a peptide hormone that allows vasodilation and plays an important role in the process of parturition. The literature suggests potential therapeutic role of H2 relaxin in PreE, however, there is a controversy on hypotensive action of the peptide. Due to the complex insulin‐like structure of relaxin (A‐ and B‐ chains, 53 amino acids, 3 disulfide bonds), a novel H2 relaxin B‐chain‐only peptide variant B7–33 (27 amino acids without any disulfide bonds) has recently been developed. This single‐chain peptide displayed equivalent efficacy to the natural H2 relaxin in several functional assays both in vitro and in vivo. Importantly, B7–33 was shown to have H2 relaxin‐like RXFP1 specific effects, particularly in endogenously expressing RXFP1 cells, thus we hypothesized that B7–33 could be an alternative and cost‐effective treatment option for PreE compared with H2 relaxin.MethodsHuman CTBs were treated with DMSO (vehicle) or 0.1, 1, 10 or 100 nM of MBG for 48 h and were co‐treated with B7–33 (25 nM) with MBG exposure, while some cells were treated with 5, 10, 25 and 50 nM B7–33 alone. Levels of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), soluble fms‐like tyrosine kinase‐1 (sFlt‐1), and soluble endoglin (sEng) were measured in culture media using ELISA kits. Cell lysates were utilized to evaluate the expression of mTOR, pAKT and MMP‐9 by western blotting. Statistical comparisons were performed using analysis of variance with Duncan's post hoc test.ResultsSecretion of sFlt‐1 and sEng were increased while VEGF and PIGF were decreased in CTBs treated with ≥1.0 nM MBG (*p < 0.05 for each). B7–33 co‐treatment significantly rescued CTBs from MBG‐induced anti‐angiogenic profile (p < 0.05 for each). There is no effect of B7–33 on sFLT‐1, sEng and PlGF; however, it increases the expression of VEGF, while CTBs were treated only with B7–33. B7–33 also causes increased expression of mTOR, pAKT and MMP‐9 in CTBs.ConclusionsB7–33 mitigates the MBG‐induced dysfunction of CTBs by attenuating anti‐angiogenic phenotype similar to that seen in PreE. This study supports the importance of continuing research of B7–33 in preE prevention.
Kinetic evaluation of the reaction of glutamate racemase with computationally derived inhibitors provides validation of a successful pharmacophore model for a new line of antibiotic therapy
Authors: Young Michael , Wells Emily , Dinsmore Olivia , Marchiano Joe , Mahfouz Tarek , Stockert Amy 
Published: November 2023

As increasing antibiotic resistance diminishes the effectiveness of once‐valuable drug therapies; research efforts must seek novel targets for new antibiotics compounds. Glutamate racemase, the enzyme responsible for the cofactor‐independent racemization of glutamic acid in bacteria, remains an untargeted step in the peptidoglycan synthesis pathway. Glutamate racemase has been shown to be essential for the viability of several pathogens including Helicobacter pylori, Escherichia coli, and Streptococcus pneumoniae, making it an ideal target for developing a new class of antibiotics. Crystal structure data for S. pneumoniae glutamate racemase is not available, thus comparative modeling was employed to develop a three‐dimensional structure for this enzyme using Prime® software. Several potential glutamate racemase inhibitors were docked into the model's active site to investigate binding modes and develop derivations to enhance for strong binding to the enzyme active site. Synthesis of the MurI gene, the gene encoding glutamate racemase, and placement into the pBluescript plasmid has allowed successful expression of functional glutamate racemase. However cellular components from the lysis of E.coli cells transformed with an inducible plasmid containing MurI have made reliable enzyme assays difficult. Addition of a His tag linker to the MurI containing plasmid allowed for purification prior to kinetic assays, thereby eliminating potential cellular contaminants that interfere with our coupled enzyme assay. Our results validated the development of a structure‐based pharmacophore model for glutamate racemase inhibitors. Previous kinetic assays have demonstrated successful inhibition with our potential drugs, but reliable determination of Ki was not possible due to interference with our coupled assay. Use of the purified enzyme for kinetic assays has removed this interference and provide more reliable and accurate Ki determination.Support or Funding InformationDepartment of Pharmaceutical and Biomedical Sciences, Ohio Northern University
It takes two to tango: The interaction of male and female genital traits mediate sperm and ejaculate transfer during mating
Authors: Friesen Christopher R. 
Published: November 2023

Genitalia are the most differentiated morphological traits in sexually reproducing organisms. Natural selection, sexual selection and sexually antagonistic coevolution contribute to genital diversification, but the relative importance of each of these evolutionary processes is unknown in most taxa, especially vertebrates. In addition, female genital morphology and behavior has been neglected in most taxa but are clearly crucial to our knowledge of genital coevolution. A critical first step toward understanding how these processes shape the evolution of specific genital traits is to determine how female and male genitalia interact during copulation. Female‐male interactions during copulation may affect sperm and ejaculate transfer in ways that influence male fitness via cryptic female choice (CFC: females biasing paternity toward or away from particular males when they mate with more than one male in a reproductive cycle). Here I present data from experimental manipulation of genitalia aimed to understand female x male interactions during copulation and how these interactions affect the fitness of both sexes. We use garter snakes as a model for genital evolution because males display great variation in form and accessory features (e.g., spines). I will also present some comparative work on garter snake genitalia, copulatory behavior, seminal fluids and future directions for that research.Support or Funding InformationNational Science Foundation Grants [IOS IOS‐0920344, IOS‐0620125, IOS‐1011727]
Resveratrol Shortens the Chronological Life Span of Saccharomyces cerevisiae under Dietary Glucose Restriction
Authors: Madrigal‐Perez Luis Alberto , Olivares‐Marin Ivanna Karina , Canizal‐Garcia Melina , Gonzalez‐Hernandez Juan Carlos , Nava Gerardo , Ramos‐Gomez Minerva 
Published: November 2023

The resveratrol (3,4′,5‐trihydroxystilbene, RSV) supplementation in mammalian systems has been related with a decrease of symptoms of diseases related with aging and the increase of longevity. However, notwithstanding the growing number of studies performed with RSV in biomedical area, the molecular mechanism by which RSV acts still remains unknown. Lately, it has been proposed that inhibition of the oxidative phosphorylation activity is the molecular mechanism of RSV action. This mechanism suggests that RSV might induce mitochondrial dysfunction resulting in oxidative damage to cells with a concomitant decrease of cell viability and cellular life span. Additionally, RSV might also have a negative impact on the ATP production via mitochondrial respiration; consequently under energy restriction the RSV effect in cellular viability could be more severe. To prove this hypothesis we measured oxygen consumption, mitochondrial membrane potential and hydrogen peroxide (H2O2) release to determine the extent of mitochondrial dysfunction, as well as the chronological life span (CLS) of Saccharomyces cerevisiae, which has been accepted as an important model of oxidative damage and aging. All the experiments were performed in a dietary glucose restriction (0.5% glucose) and with five levels of RSV (0, 10, 30, 50 and 100 μM). As expected, we found that RSV supplementation inhibits the basal respiration in all the concentrations assayed. However, we did not found any change in the mitochondrial membrane potential, except in the high‐level of RSV (100 μM) that showed an increase in this parameter. In the H2O2 release experiment, we found an inverted U‐shape dose‐response behavior, with the highest value in the 10 μM RSV and the lowest in 100 μM RSV. Altogether this data indicate that 100 μM RSV might produce a mitochondrial dysfunction, since inhibits the oxygen consumption and increases the mitochondrial membrane potential. Finally, as anticipated, we found that supplementation of S. cerevisiae cultures with 100 μM RSV decreased CLS. Collectively, this data supports the hypothesis that RSV supplementation decreases CLS in a dietary glucose restriction condition as a result of mitochondrial dysfunction.Support or Funding InformationLuis Alberto Madrigal‐Perez (PRODEP scholarship ITESCH‐002).
Teniposide reduces vascular calcification by inhibiting BMP2 expression
Authors: Liu Lipei , Zhang Xiaomeng , Chen Yuanli , Duan Yajun , Han Jihong 
Published: November 2023

Vascular calcification, characterized by ectopic calcium deposition, is clinically associated with the development of chronic kidney disease, atherosclerosis and diabetes mellitus. Bone morphogenetic proteins 2 (BMP2) regulates orthotopic bone formation physiologically. However, high expression of BMP2 in atherosclerotic lesions suggests that BMP2 may play an important role in vascular calcification under pathological conditions. Teniposide, a well‐defined DNA topoisomerase II (Topo II) inhibitor, is widely prescribed to patients with various types of cancers. In addition, we previously reported that teniposide is able to activate macrophage ABCA1 expression and free cholesterol efflux, and hepatic CETP expression indicating that Top II inhibitors may demonstrate multiple anti‐atherogenic properties. Herein, we investigated if teniposide can influence calcification in aortic root, and BMP2 is involved in this process. In vitro, we employed the β‐glycerophosphoric acid method to induce calcification in human aortic smooth muscle cells (HASMCs). We determined that treatment of HASMCs with teniposide attenuated the phosphate‐induced calcium deposition. Mechanistically, we found that teniposide suppressed HASMC calcification by decreasing the BMP2 and Smad1/5/8 signaling cascade in a p53‐dependent manner. To determine the effect of teniposide on vascular calcification, apoE deficient (apoE−/−) mice were randomly divided into 3 groups and received the following treatment: Group 1 (control), mice were fed high‐fat diet (HFD) for 4 months; Group 2 (prevention group), mice were fed HFD containing teniposide [0.5 mg/kg bodyweight/day, (mpk)]; and Group 3 (therapy group) mice were pre‐fed with HFD alone for 3 months followed by one month of HFD containing teniposide (0.5 mpk). At the end of experiment, we determined the development of lesions and calcification. We observed that teniposide reduced atherosclerotic lesions and vascular calcification in lesion areas, in both prevention and therapy groups. The immunofluorescent staining demonstrates that BMP2 expression in the calcified vascular areas was also inhibited by teniposide. In contrast, the results of microCT analysis on the distal femur show no effect of teniposide on bone structure indicating the tissue‐type specific function of teniposide. Taken together, our study demonstrated that teniposide treatment ameliorates vascular calcification both in vitro and in vivo which was completed by regulating p53‐BMP2 pathway. Our study also demonstrates another biological function of TopII inhibitors.Support or Funding InformationThe National Science Foundation of China Grants 81473204 to J Han and 81573427 to Y Duan
Hepatic Metabolic Reprogramming by Caloric Restriction in Rhesus Monkeys
Authors: Rhoads Timothy W. , Burhans Maggie S. , McIlwain Sean J. , Hutchins Paul D. , Chen Vincent B. , Eghbalnia Hamid R. , Ong Irene M. , Markley John L. , Coon Joshua J. , Anderson Rozalyn M. 
Published: November 2023

Caloric Restriction (CR) without malnutrition delays aging and the incidence of age‐related diseases. Adult‐onset CR enhances survival and reduces age‐related mortality in rhesus monkey (Macaca mulatta) compared to ad libitum fed controls, suggesting that the longevity benefits of CR can apply to primates, including humans; however, the mechanisms responsible for the longevity benefits of CR are to date unknown. We used five high‐throughput, high‐resolution techniques to generate hepatic molecular profiles for monkeys at baseline and 2 years after introduction of the CR diet. Approximately 20,000 biological molecules were detected and quantified across two time points, enabling us to determine the trajectory of response to CR. We find that CR upregulated many metabolic pathways, including fatty acid metabolism, the citric acid cycle, and branched chain amino acid degradation, and downregulated immune and inflammation pathways at the transcriptional and proteomic levels. Our data are suggestive of a novel role for the processing of mRNA in the mechanisms of CR. We report significant remodeling of the non‐histone acetylome, especially on proteins localized to the mitochondria. Hepatic lipid composition analysis showed increased levels of highly unsaturated trigylcerides and increased prevalence of odd‐chain phospholipids as a result of CR, while metabolomics analysis revealed increases in aspartate and 3‐hydroxybutyrate, the first ketone body to be synthesized under fasting conditions. These data reveal that CR activates a distinct hepatic metabolic program, suggesting that the metabolic adaptation to CR sets the stage for downstream longevity benefits.Support or Funding InformationThis work was supported by NIH grants R01AG040178, R56AG047358, and R01AG037000.
Development and standardization of methodology for diagnosis and detection of response to chemotherapy in patients with colorectal cancer
Authors: Assis Jéssica Vieira , Coutinho Lucélia , Oliveira Matheus , Cruz Renata , Grenfell Rafaella 
Published: November 2023

Colorectal cancer (CRC) is a heterogeneous disease. Subtyping in gene expression is recognized as a relevant criterion for classification of the disease, but its clinical usefulness is hampered by the lack of standard and methodology to be used between the clinical studies and analyze the responsiveness of each patient to the chemotherapeutic treatment. CXCR4 (CD184) when highly expressed generates proliferation and cell survival, angiogenesis and metastatic tumor migration. When the CD26 gene is less expressed, there is a decrease in local chemokine levels, thereby controlling the tumor in situ. Patients responsive to 5‐FU, FOLFOX and FOLFIRI present a significant reduction in CXCR4 expression and progressive increase of CD26, where the inverse ratio is seen in nonresponsive patients. Thus, in order to standardize high sensitivity methodology for diagnosis and prognosis, we developed the use of technology of detection of circulating antigens through the use of nanotechnology associated with antibodies. Thus, following in vitro methodology, anti‐CXCR4 (CD184) and anti‐CD26 pAbs were due to producing clones of IgM or IgG subtypes for each marker, since the difference between the expressions of such genes suggests an interesting imbalance, generating a possible response. After standardization of tumor induction in nude mice, HT‐29 cells (mutation at codon 273 of the p53 gene) were performed, four inoculations of these proteins were carried out and throughout the production process, immunoenzymatic assays were performed for identification and selection of clones Positive. The purified antibodies were tested to SDS‐PAGE and Western Blotting for confirmation of specificity. Thereafter, the HT‐29 cells removed from mice will be labeled to verify the expression of CXCR4 and CD26 on their surfaces from the inoculated pAb and thereafter, tissues from patients at different stages of the disease will also be labeled. We thus sought, in clinical cases, to assist the medical team in the decision of the treatment leading the patient to the surgical intervention and/or the conventional chemotherapies.Support or Funding InformationFinancial support: Fiocruz, PDTIS, FINEP (Brazil).
MED28 Mediates Glucose Metabolism in Human Colorectal Cancer Cells
Authors: Hsieh Nien‐Tsu , Huang Chun‐Yin , Weng Yu‐Ting , Lin Yi‐Chang , Lee Ming‐Fen 
Published: November 2023

Most normal cells mainly rely on mitochondrial oxidative phosphorylation to generate energy, whereas cancer cells preferentially rely on aerobic glycolysis or so‐called “Warburg effect” even in the presence of sufficient oxygen. Albeit inefficient energy production, cancer cells compensate for this disadvantage by accelerating the uptake of glucose and increasing the rate of glycolysis. To increase the rate of glycolysis also provides the glycolytic intermediates for the synthesis of macromolecules, DNA, protein, and lipid, to keep up with the rapid proliferation mode. Recent studies have shown that Mediator Complex is involved in human diseases, including cancer, cardiovascular disease, metabolic disorder, and neurological disease. MED28, a Mediator subunit for transcriptional activation, is also an interactor with multiple proteins including Grb2, Src kinases, actin cytoskeleton, and merlin. MED28 was found highly expressed in colorectal cancer. In addition, our laboratory has previously reported that MED28 mediates Wnt/β‐catenin signaling in human colorectal cancer cells. This study is to investigate if MED28 plays a role in cell growth in the aspect of glucose metabolism in colorectal cancer. Expression profiling upon MED28 knockdown in SW480 colorectal cancer cells showed that MED28 suppression down‐regulated genes involved in glucose metabolism including glucose transporters and glycolytic enzymes, and the effect on several proteins and enzymes have been confirmed at both transcriptional and translational levels by RT‐PCR and Western blotting, respectively. The underlying mechanism is currently under investigation. In summary, MED28 regulates cell growth, at least in part, by mediating Warburg effect in colorectal cancer.Support or Funding InformationThis work was supported by the grants NSC102‐2320‐B‐309‐001‐MY3 and MOST105‐2320‐B‐309‐001 to M‐F Lee.