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1.
Med Phys ; 50(12): 8057-8062, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37655886

ABSTRACT

PURPOSE: Transmission-target x-ray tubes generate more x-rays than reflection thick-target x-ray tubes. A transmission x-ray tube combined with radiosensitizers has a better radiation enhancement effect. This study investigated the feasibility of using a transmission x-ray tube with radiosensitizers in clinical radiotherapy and its effect on radiation dose enhancement. METHODS: This study used MCNP6.2 to simulate the model of a transmission x-ray tube and Co-60 beam.   The radiation enhancement effect of radiosensitizers was examined with iodine-127 (I-127), radioiodinated iododeoxyuridine (IUdR), and gold nanoparticles (GNPs). RESULTS: The study results showed that the dose enhancement factor (DEF) of the transmission x-ray tube with GNPs was 10.27, which was higher than that of I-127 (6.46) and IUdR (3.08). The DEF of the Co-60 beam with GNPs, I-127, and IUdR was 1.23, 1.19, and 1.2, respectively. The Auger electron flux of the transmission x-ray tube with GNPs was 1.19E+05 particles/cm2 . CONCLUSIONS: This study found that a transmission x-ray tube with appropriate radiosensitizers could produce a high rate of Auger electrons to fulfill the radiation enhancement effect, and this procedure has the potential to become a radiotherapy modality.


Subject(s)
Metal Nanoparticles , Radiation-Sensitizing Agents , Idoxuridine , X-Rays , Monte Carlo Method , Gold , Metal Nanoparticles/therapeutic use
2.
Chemistry ; 29(30): e202300017, 2023 May 26.
Article in English | MEDLINE | ID: mdl-36880483

ABSTRACT

The development of a universal approach for precisely tuning the electrochemical characteristics of conducting carbon nanotubes for tracking harmful agents in the human body with high selectivity and sensitivity remains a challenge. Herein, we describe a simplistic, versatile, and general approach to the construction of functionalized electrochemical material. The design of electrochemical material consists of (i) modification of multiwalled carbon nanotubes (MWCNT) with dipodal naphthyl-based dipodal urea (KR-1) through non-covalent functionalization (KR-1@MWCNT) which enhances the dispersibility of MWCNT and hence conductivity, (ii) complexation of KR-1@MWCNT with Hg2+ accelerate the electron transfer in the material which amplify the detection response of functionalized material (i. e., Hg/KR-1@MWCNT) towards various thymidine analogues. Further, the application of functionalized electrochemical material (Hg/KR-1@MWCNT) achieves real-time electrochemical monitoring of harmful antiviral drug 5-iodo-2'-iododeoxyuridine (IUdR) levels in human serum for the first time.


Subject(s)
Nanotubes, Carbon , Humans , Nanotubes, Carbon/chemistry , Antiviral Agents , Electrochemical Techniques , Idoxuridine
3.
Curr Protoc ; 2(7): e502, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35895086

ABSTRACT

Modification of nucleosides via cross-coupling processes has been carried out extensively on unprotected halonucleosides to produce functionalized nucleosides that are often developed for incorporation into oligonucleotides or used as fluorescent probes. This approach requires protection of the 5'-OH with the 4,4'-dimethoxytrityl (DMTr) group, which is complicated and a common cause of reaction failure. Here we report a method for direct functionalization of 5'-O-DMTr-5-iodo-2'-deoxyuridine via Suzuki-Miyaura cross-coupling, Heck alkenylation, and carboamidation. This approach facilitates rapid synthesis of a variety of C5-substituted 5'-O-DMTr-2'-deoxyuridine derivatives. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Synthesis of the SerrKap palladacycle complex Basic Protocol 2: Suzuki-Miyaura coupling of 5'-O-DMTr-5-iodo-2'-deoxyuridine using SerrKap palladacycle Basic Protocol 3: Heck coupling of 5'-O-DMTr-5-iodo-2'-deoxyuridine using SerrKap palladacycle Basic Protocol 4: Heck coupling of 5'-O-DMTr-5-iodo-2'-deoxyuridine with Ruth linker using Pd(OAc)2 /PTABS Basic Protocol 5: Carbonylative amidation of 5'-O-DMTr-5-iodo-2'-deoxyuridine using Pd(OAc)2 /PTABS.


Subject(s)
Idoxuridine , Palladium , Catalysis , Nucleosides , Oligonucleotides
4.
Mol Divers ; 26(5): 2631-2645, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35001230

ABSTRACT

Coronavirus disease 2019 (COVID-19) is caused by novel severe acute respiratory syndrome coronavirus (SARS-CoV-2). Its main protease, 3C-like protease (3CLpro), is an attractive target for drug design, due to its importance in virus replication. The analysis of the radial distribution function of 159 3CLpro structures reveals a high similarity index. A study of the catalytic pocket of 3CLpro with bound inhibitors reveals that the influence of the inhibitors is local, perturbing dominantly only residues in the active pocket. A machine learning based model with high predictive ability against SARS-CoV-2 3CLpro is designed and validated. The model is used to perform a drug-repurposing study, with the main aim to identify existing drugs with the highest 3CLpro inhibition power. Among antiviral agents, lopinavir, idoxuridine, paritaprevir, and favipiravir showed the highest inhibition potential. Enzyme - ligand interactions as a key ingredient for successful drug design.


Subject(s)
COVID-19 Drug Treatment , SARS-CoV-2 , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Catalytic Domain , Coronavirus 3C Proteases , Drug Repositioning , Humans , Idoxuridine , Ligands , Lopinavir , Molecular Docking Simulation , Peptide Hydrolases/metabolism , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology
5.
J Vis Exp ; (176)2021 10 22.
Article in English | MEDLINE | ID: mdl-34747407

ABSTRACT

The regulation of cell cycle phase is an important aspect of cellular proliferation and homeostasis. Disruption of the regulatory mechanisms governing the cell cycle is a feature of a number of diseases, including cancer. Study of the cell cycle necessitates the ability to define the number of cells in each portion of cell cycle progression as well as to clearly delineate between each cell cycle phase. The advent of mass cytometry (MCM) provides tremendous potential for high throughput single cell analysis through direct measurements of elemental isotopes, and the development of a method to measure the cell cycle state by MCM further extends the utility of MCM. Here we describe a method that directly measures 5-iodo-2'-deoxyuridine (IdU), similar to 5-bromo-2´-deoxyuridine (BrdU), in an MCM system. Use of this IdU-based MCM provides several advantages. First, IdU is rapidly incorporated into DNA during its synthesis, allowing reliable measurement of cells in the S-phase with incubations as short as 10-15 minutes. Second, IdU is measured without the need for secondary antibodies or the need for DNA degradation. Third, IdU staining can be easily combined with measurement of cyclin B1, phosphorylated retinoblastoma protein (pRb), and phosphorylated histone H3 (pHH3), which collectively provides clear delineation of the five cell cycle phases. Combination of these cell cycle markers with the high number of parameters possible with MCM allow combination with numerous other metrics.


Subject(s)
Idoxuridine , Bromodeoxyuridine/metabolism , Cell Cycle , Flow Cytometry/methods , Idoxuridine/metabolism , Staining and Labeling
6.
Science ; 373(6557)2021 08 20.
Article in English | MEDLINE | ID: mdl-34301855

ABSTRACT

Stochastic fluctuations in gene expression ("noise") are often considered detrimental, but fluctuations can also be exploited for benefit (e.g., dither). We show here that DNA base excision repair amplifies transcriptional noise to facilitate cellular reprogramming. Specifically, the DNA repair protein Apex1, which recognizes both naturally occurring and unnatural base modifications, amplifies expression noise while homeostatically maintaining mean expression levels. This amplified expression noise originates from shorter-duration, higher-intensity transcriptional bursts generated by Apex1-mediated DNA supercoiling. The remodeling of DNA topology first impedes and then accelerates transcription to maintain mean levels. This mechanism, which we refer to as "discordant transcription through repair" ("DiThR," which is pronounced "dither"), potentiates cellular reprogramming and differentiation. Our study reveals a potential functional role for transcriptional fluctuations mediated by DNA base modifications in embryonic development and disease.


Subject(s)
Cell Differentiation , Cellular Reprogramming , DNA Repair , DNA-(Apurinic or Apyrimidinic Site) Lyase/metabolism , DNA/chemistry , Gene Expression , Transcription, Genetic , Animals , Cells, Cultured , Computer Simulation , DNA/genetics , DNA/metabolism , Embryonic Stem Cells , Gene Expression/drug effects , Idoxuridine/metabolism , Idoxuridine/pharmacology , Mice , Models, Genetic , Nanog Homeobox Protein/genetics , Nucleic Acid Conformation , RNA, Messenger/genetics , RNA, Messenger/metabolism , Single-Cell Analysis , Stochastic Processes , Thymidine Kinase/genetics , Thymidine Kinase/metabolism , Transcription, Genetic/drug effects
7.
J Photochem Photobiol B ; 205: 111827, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32120183

ABSTRACT

5-iodo-2-deoxyuridine (IUdR) has been demonstrated to induce an appreciable radiosensitizing effect on glioblastoma patients, but due to the short circulation half-life times and failure to pass through the blood-brain barrier (BBB), its clinical use is limited. Accordingly, in this study, we used magnetic graphene oxide (NGO/SPIONs) nanoparticles coated with PLGA polymer as a dynamic nanocarrier for IUdR and, evaluated its sensitizing enhancement ratio in combination with a single dose X-ray at clinically megavoltage energies for treatment of C6 glioma rats. Nanoparticles were characterized using Zetasizer and TEM microscopy, and in vitro biocompatibility of nanoparticles was assessed with MTT assay. IUdR/MNPs were intravenously administered under a magnetic field (1.3 T) on day 13 after the implantation of C6 cells. After a day following the injection, rats exposed with radiation (8 Gy). ICP-OES analysis data indicated an effective magnetic targeting, leading to remarkably improved penetration through the BBB. In vivo release analysis with HPLC indicated sustained release of IUdR and, prolonged the lifespan in plasma (P < .01). In addition, our findings revealed a synergistic effect for IUdR/MNPs coupled with radiation, which significantly inhibited the tumor expansion (>100%), prolonged the survival time (>100%) and suppressed the anti-apoptotic response of glioma rats by increasing Bax/Bcl-2 ratio (2.13-fold) in compared with the radiation-only. In conclusion, besides high accumulation in targeted tumor sites, the newly developed IUdR/MNPs, also exhibited the ability of IUdR/MNPs to significantly enhance radiosensitizing effect, improve therapeutic efficacy and increase toxicity for glioma-bearing rats.


Subject(s)
Brain Neoplasms/drug therapy , Glioma/drug therapy , Graphite/administration & dosage , Idoxuridine/administration & dosage , Nanoparticles/administration & dosage , Polylactic Acid-Polyglycolic Acid Copolymer/administration & dosage , Radiation-Sensitizing Agents/administration & dosage , Animals , Brain/metabolism , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Cell Line, Tumor , Drug Liberation , Glioma/metabolism , Glioma/pathology , Graphite/chemistry , Graphite/pharmacokinetics , Hydrogen-Ion Concentration , Idoxuridine/pharmacokinetics , Magnetic Phenomena , Male , Nanoparticles/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer/pharmacokinetics , Rabbits , Radiation-Sensitizing Agents/pharmacokinetics , Rats, Wistar , Tumor Burden/drug effects
9.
Clin Cancer Res ; 25(20): 6035-6043, 2019 10 15.
Article in English | MEDLINE | ID: mdl-31337643

ABSTRACT

PURPOSE: Iododeoxyuridine (IUdR) is a potent radiosensitizer; however, its clinical utility is limited by dose-limiting systemic toxicities and the need for prolonged continuous infusion. 5-Iodo-2-pyrimidinone-2'-deoxyribose (IPdR) is an oral prodrug of IUdR that, compared with IUdR, is easier to administer and less toxic, with a more favorable therapeutic index in preclinical studies. Here, we report the clinical and pharmacologic results of a first-in-human phase I dose escalation study of IPdR + concurrent radiation therapy (RT) in patients with advanced metastatic gastrointestinal (GI) cancers. PATIENTS AND METHODS: Adult patients with metastatic GI cancers referred for palliative RT to the chest, abdomen, or pelvis were eligible for study. Patients received IPdR orally once every day × 28 days beginning 7 days before the initiation of RT (37.5 Gy in 2.5 Gy × 15 fractions). A 2-part dose escalation scheme was used, pharmacokinetic studies were performed at multiple time points, and all patients were assessed for toxicity and response to Day 56. RESULTS: Nineteen patients were entered on study. Dose-limiting toxicity was encountered at 1,800 mg every day, and the recommended phase II dose is 1,200 mg every day. Pharmacokinetic analyses demonstrated achievable and sustainable levels of plasma IUdR ≥1 µmol/L (levels previously shown to mediate radiosensitization). Two complete, 3 partial, and 9 stable responses were achieved in target lesions. CONCLUSIONS: Administration of IPdR orally every day × 28 days with RT is feasible and tolerable at doses that produce plasma IUdR levels ≥1 µmol/L. These results support the investigation of IPdR + RT in phase II studies.


Subject(s)
Chemoradiotherapy/methods , Gastrointestinal Neoplasms/therapy , Idoxuridine/pharmacokinetics , Pyrimidine Nucleosides/administration & dosage , Radiation-Sensitizing Agents/administration & dosage , Administration, Oral , Adult , Aged , Aged, 80 and over , Dose Fractionation, Radiation , Feasibility Studies , Female , Gastrointestinal Neoplasms/pathology , Humans , Idoxuridine/administration & dosage , Idoxuridine/toxicity , Male , Maximum Tolerated Dose , Middle Aged , Neoplasm Staging , Prodrugs/administration & dosage , Prodrugs/pharmacokinetics , Prodrugs/toxicity , Pyrimidine Nucleosides/pharmacokinetics , Pyrimidine Nucleosides/toxicity , Radiation-Sensitizing Agents/pharmacokinetics , Radiation-Sensitizing Agents/toxicity , Treatment Outcome
10.
Cytometry A ; 95(10): 1075-1084, 2019 10.
Article in English | MEDLINE | ID: mdl-31150166

ABSTRACT

We present a new method to directly quantify the dynamics of differentiation of multiple cellular subsets in unperturbed mice. We combine a pulse-chase protocol of 5-iodo-2'-deoxyuridine (IdU) injections with subsequent analysis by mass cytometry (CyTOF) and mathematical modeling of the IdU dynamics. Measurements by CyTOF allow for a wide range of cells to be analyzed at once, due to the availability of a large staining panel without the complication of fluorescence spillover. These are also compatible with direct detection of integrated iodine signal, with minimal impact on immunophenotyping based on the surface markers. Mathematical modeling beyond a binary classification of surface marker abundance allows for a continuum of cellular states as the cells transition from one state to another. Thus, we present a complete and robust method for directly quantifying differentiation at the systemic level, allowing for system-wide comparisons between different mouse strains and/or experimental conditions. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.


Subject(s)
Flow Cytometry/methods , Hematopoiesis , Idoxuridine/metabolism , Models, Theoretical , Animals , B-Lymphocytes/cytology , Cell Differentiation , Female , Mice, Inbred C57BL , Neutrophils/cytology , Phenotype , Time Factors
11.
Methods Mol Biol ; 1989: 193-215, 2019.
Article in English | MEDLINE | ID: mdl-31077107

ABSTRACT

The regulated proliferation of cells is a critical factor in tumor progression, antineoplastic therapies, immune system regulation, and the cellular developmental of multicellular organisms. While measurement of cell cycle state by fluorescent flow cytometry is well established, mass cytometry allows the cell cycle to be measured along with large numbers of other antigens enabling characterization of the complex interactions between the cell cycle and wide variety of cellular processes. This method describes the use of mass cytometry for the analysis of cell cycle state for cells from three different sources: in vitro cultured cell lines, ex vivo human blood or bone marrow, and in vivo labeling of murine tissues. The method utilizes incorporation of 5-Iodo-2'-deoxyuridine (IdU), combined with measurement of phosphorylated retinoblastoma protein (pRb), Cyclin B1, and phosphorylated Histone H3 (pHH3). These measurements can be integrated into a gating strategy that enables clear separation of all five phases of the cell cycle.


Subject(s)
Cell Cycle , Cyclin B1/analysis , Flow Cytometry/methods , Histones/analysis , Mass Spectrometry/methods , Retinoblastoma Protein/analysis , Staining and Labeling/methods , Animals , Bone Marrow Cells/metabolism , Humans , Idoxuridine/analogs & derivatives , Idoxuridine/metabolism , Mice , Phosphorylation
12.
Methods Mol Biol ; 1999: 307-318, 2019.
Article in English | MEDLINE | ID: mdl-31127587

ABSTRACT

Understanding the molecular dynamics of DNA replication in vivo has been a formidable challenge requiring the development of advanced technologies. Over the past 50 years or so, studies involving DNA autoradiography in bacterial cells have led to sophisticated DNA tract analyses in human cells to characterize replication dynamics at the single-molecule level. Our own lab has used DNA fiber analysis to characterize replication in helicase-deficient human cells. This work led us to propose a model in which the human DNA helicase RECQ1 acts as a governor of the single-stranded DNA binding protein RPA and regulates its bioavailability for DNA synthesis. We have also used the DNA fiber approach to investigate the interactive role of DDX11 helicase with a replication fork protection protein (Timeless) in human cells when they are under pharmacologically induced stress. In this methods chapter, we present a step-by-step protocol for the single-molecule DNA fiber assay. We describe experimental designs to study replication stress and staining patterns from pulse-chase labeling experiments to address the dynamics of replication forks in stressed cells.


Subject(s)
DNA Damage/genetics , DNA Replication/genetics , Single Molecule Imaging/methods , Cell Cycle Proteins/metabolism , DEAD-box RNA Helicases/metabolism , DNA Damage/drug effects , DNA Helicases/metabolism , DNA Replication/drug effects , DNA, Single-Stranded/metabolism , Deoxyuridine/analogs & derivatives , Deoxyuridine/toxicity , HeLa Cells , Humans , Idoxuridine/analogs & derivatives , Idoxuridine/toxicity , Intracellular Signaling Peptides and Proteins/metabolism , RecQ Helicases/metabolism , Replication Protein A/metabolism
13.
Methods Mol Biol ; 1999: 319-325, 2019.
Article in English | MEDLINE | ID: mdl-31127588

ABSTRACT

The ability to analyze individual DNA fibers undergoing active DNA synthesis has emerged as a powerful technique in the field of DNA replication. Much of the initial analysis has focused on replication throughout the genome. However, more recent advancements in this technique have allowed for the visualization of replication patterns at distinct loci or regions within the genome. This type of locus-specific resolution will greatly enhance our understanding of the dynamics of DNA replication in regions that provide a challenge to the replication machinery. Here, we describe a protocol that will allow for the visualization of DNA replication through one of the most structurally complex regions in the human genome, the telomeric DNA.


Subject(s)
DNA Replication , DNA/genetics , In Situ Hybridization, Fluorescence/methods , Molecular Imaging/methods , Telomere/metabolism , Cell Line , DNA/chemistry , Deoxyuridine/analogs & derivatives , Deoxyuridine/chemistry , Fluorescent Antibody Technique, Direct/methods , Genetic Loci , Humans , Idoxuridine/chemistry , Microscopy, Fluorescence/instrumentation , Microscopy, Fluorescence/methods , Molecular Imaging/instrumentation , Molecular Probes/chemistry , Staining and Labeling/methods
14.
Sci Rep ; 9(1): 5447, 2019 04 01.
Article in English | MEDLINE | ID: mdl-30931986

ABSTRACT

Heterotopic ossification (HO) is a pathological condition characterized by the deposition of mineralized tissue in ectopic locations such as the skeletal muscle. The precise cellular origin and molecular mechanisms underlying HO are still debated. In our study we focus on the differentiation of mesoangioblasts (MABs), a population of multipotent skeletal muscle precursors. High-content screening for small molecules that perturb MAB differentiation decisions identified Idoxuridine (IdU), an antiviral and radiotherapy adjuvant, as a molecule that promotes MAB osteogenic differentiation while inhibiting myogenesis. IdU-dependent osteogenesis does not rely on the canonical BMP-2/SMADs osteogenic pathway. At pro-osteogenic conditions IdU induces a mild DNA Damage Response (DDR) that activates ATM and p38 eventually promoting the phosphorylation of the osteogenesis master regulator RUNX2. By interfering with this pathway IdU-induced osteogenesis is severely impaired. Overall, our study suggests that induction of the DDR promotes osteogenesis in muscle resident MABs thereby offering a new mechanism that may be involved in the ectopic deposition of mineralized tissue in the muscle.


Subject(s)
Cell Differentiation , DNA Damage , Multipotent Stem Cells/pathology , Muscle, Skeletal/pathology , Osteogenesis , Animals , Bone Morphogenetic Protein 2/metabolism , Cell Differentiation/drug effects , Core Binding Factor Alpha 1 Subunit/metabolism , Idoxuridine/pharmacology , Multipotent Stem Cells/drug effects , Muscle, Skeletal/drug effects , Ossification, Heterotopic , Smad Proteins/metabolism
15.
Chemistry ; 25(7): 1773-1780, 2019 Feb 01.
Article in English | MEDLINE | ID: mdl-30398293

ABSTRACT

Halogenated nucleosides, such as 5-iodo-2'-deoxyuridine and 5-iodo-2'-deoxycytidine, are incorporated into the DNA of replicating cells to facilitate DNA single-strand breaks and intra- or interstrand crosslinks upon UV irradiation. In this work, it is shown that the naphthyl-based organoselenium compounds can mediate the dehalogenation of halogenated pyrimidine-based nucleosides, such as 5-X-2'-deoxyuridine and 5-X-2'-deoxycytidine (X=Br or I). The rate of deiodination was found to be significantly higher than that of the debromination for both nucleosides. Furthermore, the deiodination of iodo-cytidines was found to be faster than that of iodo-uridines. The initial rates of the deiodinations of 5-iodocytosine and 5-iodouracil indicated that the nature of the sugar moiety influences the kinetics of the deiodination. For both the nucleobases and nucleosides, the deiodination and debromination reactions follow a halogen-bond-mediated and addition/elimination pathway, respectively.


Subject(s)
Nucleosides/chemistry , Organoselenium Compounds/chemistry , Crystallography, X-Ray , Halogenation , Idoxuridine/analogs & derivatives , Idoxuridine/chemistry , Magnetic Resonance Spectroscopy , Molecular Conformation
16.
Int J Radiat Biol ; 94(11): 1027-1037, 2018 11.
Article in English | MEDLINE | ID: mdl-29985733

ABSTRACT

PURPOSE: Glioblastoma multiform (GBM) is the most prevalent and aggressive type of primary brain tumor. None of the current conventional treatment methods has improved treatment considerably. Therefore, in this study the effect of magnetic nanoparticles coated with poly (caprolactone)-poly (ethylene glycol) (PCL-PEG) as a 5-iodo 2'deoxyuridine (IUdR) carrier in the presence of hyperthermia and 6 MV (megavoltage) X-ray radiation, were investigated in a spheroid model of U87MG glioblastoma cell line using colony formation assay. MATERIALS AND METHODS: First, the human glioblastoma cell line U87MG was cultured as a spheroid using the liquid overlay technique. Spheroids on day 10 with 100 mm diameters were treated with 1 µM IUdR or nanoparticles as IUdR carriers for one volume doubling time (VDT) of spheroids (67 h) and hyperthermia at 43 °C for 1 h, and then irradiated with 2 Gy of 6 MV X-ray in different groups. Finally, the effect of treatment on colony-forming ability was obtained by colony formation and alkaline assay. RESULTS: Our results revealed that hyperthermia in combination with radiation could significantly reduce the colony number of glioblastoma spheroid cells treated with IUdR or nanoparticles as IUdR carriers. However, the extent of reduction in colony number following treatment with IUdR-loaded nanoparticles in combination with hyperthermia and then X-ray radiation was significantly more than free IUdR. CONCLUSION: According to this study, PCL-PEG-coated magnetic nanoparticles are effective delivery vehicles for IUdR into cells. Moreover, they can act as a radiosensitizer and thermosensitizer in the treatment of the glioblastoma cell line.


Subject(s)
Drug Carriers/chemistry , Glioblastoma/pathology , Hyperthermia, Induced , Idoxuridine/analogs & derivatives , Magnetite Nanoparticles/chemistry , Polyesters/chemistry , Polyethylene Glycols/chemistry , Biological Transport , Cell Line, Tumor , Cell Proliferation/radiation effects , Combined Modality Therapy , Drug Carriers/metabolism , Glioblastoma/radiotherapy , Humans , Idoxuridine/chemistry , Particle Size , Spheroids, Cellular/pathology , Spheroids, Cellular/radiation effects
17.
Chem Commun (Camb) ; 54(57): 8003-8006, 2018 Jul 12.
Article in English | MEDLINE | ID: mdl-29967912

ABSTRACT

The on-column functionalization of oligodeoxynucleotides via base-free Suzuki cross-coupling reactions is reported herein. These cross-coupling reactions were carried out with various boronic acids and either full-length modified oligonucleotides containing one or more 2'-deoxy-5-iodouridine (5IdU) monomer(s) or on oligonucleotide fragments immediately after incorporation of 5IdU. Five different functionalities were coupled to oligonucleotides containing one or three attachment points.


Subject(s)
Oligodeoxyribonucleotides/chemistry , Boronic Acids/chemistry , Catalysis , Idoxuridine/analogs & derivatives , Idoxuridine/chemistry , Palladium/chemistry
18.
J Radiat Res ; 59(4): 411-429, 2018 Jul 01.
Article in English | MEDLINE | ID: mdl-29800458

ABSTRACT

Gene expression analysis was carried out in Jurkat cells in order to identify candidate genes showing significant gene expression alterations allowing robust discrimination of the Auger emitter 123I, incorporated into the DNA as 123I-iododeoxyuridine (123IUdR), from α- and γ-radiation. The γ-H2AX foci assay was used to determine equi-effect doses or activity, and gene expression analysis was carried out at similar levels of foci induction. Comparative gene expression analysis was performed employing whole human genome DNA microarrays. Candidate genes had to show significant expression changes and no altered gene regulation or opposite regulation after exposure to the radiation quality to be compared. The gene expression of all candidate genes was validated by quantitative real-time PCR. The functional categorization of significantly deregulated genes revealed that chromatin organization and apoptosis were generally affected. After exposure to 123IUdR, α-particles and γ-rays, at equi-effect doses/activity, 155, 316 and 982 genes were exclusively regulated, respectively. Applying the stringent requirements for candidate genes, four (PPP1R14C, TNFAIP8L1, DNAJC1 and PRTFDC1), one (KLF10) and one (TNFAIP8L1) gene(s) were identified, respectively allowing reliable discrimination between γ- and 123IUdR exposure, γ- and α-radiation, and α- and 123IUdR exposure, respectively. The Auger emitter 123I induced specific gene expression patterns in Jurkat cells when compared with γ- and α-irradiation, suggesting a unique cellular response after 123IUdR exposure. Gene expression analysis might be an effective tool for identifying biomarkers for discriminating different radiation qualities and, furthermore, might help to explain the varying biological effectiveness at the mechanistic level.


Subject(s)
Alpha Particles , Biomarkers/metabolism , Gamma Rays , Gene Expression Profiling , Idoxuridine/pharmacology , DNA/metabolism , DNA Damage , Down-Regulation/drug effects , Down-Regulation/genetics , Down-Regulation/radiation effects , Genetic Association Studies , Histones/metabolism , Humans , Jurkat Cells , Signal Transduction/genetics , Up-Regulation/drug effects , Up-Regulation/genetics , Up-Regulation/radiation effects
19.
Nucleic Acids Res ; 46(11): e65, 2018 06 20.
Article in English | MEDLINE | ID: mdl-29546376

ABSTRACT

Pd-catalyzed C-C bond formation, an important vertebra in the spine of synthetic chemistry, is emerging as a valuable chemoselective transformation for post-synthetic functionalization of biomacromolecules. While methods are available for labeling protein and DNA, development of an analogous procedure to label RNA by cross-coupling reactions remains a major challenge. Herein, we describe a new Pd-mediated RNA oligonucleotide (ON) labeling method that involves post-transcriptional functionalization of iodouridine-labeled RNA transcripts by using Suzuki-Miyaura cross-coupling reaction. 5-Iodouridine triphosphate (IUTP) is efficiently incorporated into RNA ONs at one or more sites by T7 RNA polymerase. Further, using a catalytic system made of Pd(OAc)2 and 2-aminopyrimidine-4,6-diol (ADHP) or dimethylamino-substituted ADHP (DMADHP), we established a modular method to functionalize iodouridine-labeled RNA ONs in the presence of various boronic acid and ester substrates under very mild conditions (37°C and pH 8.5). This method is highly chemoselective, and offers direct access to RNA ONs labeled with commonly used fluorescent and affinity tags and new fluorogenic environment-sensitive nucleoside probes in a ligand-controlled stereoselective fashion. Taken together, this simple approach of generating functional RNA ON probes by Suzuki-Miyaura coupling will be a very important addition to the resources and tools available for analyzing RNA motifs.


Subject(s)
Oligonucleotides/chemistry , RNA Probes/chemistry , RNA/chemistry , Staining and Labeling/methods , Boronic Acids/chemistry , Catalysis , DNA-Directed RNA Polymerases/metabolism , Idoxuridine/analogs & derivatives , Idoxuridine/chemistry , Molecular Structure , Palladium/chemistry , Pyrimidines/chemistry , Viral Proteins/metabolism
20.
J Eur Acad Dermatol Venereol ; 32(4): 537-541, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29125649

ABSTRACT

Milker's nodule virus, also called paravaccinia virus, is a DNA virus of the parapoxvirus genus transmitted from infected cows to humans. It results from contact with cattle, cattle by-products or fomites. Classified as an occupational disorder, those at risk of exposure include farmers, butchers and agricultural tourists. The viral infection begins 5-15 days after inoculation as an erythematous-purple, round nodule with a clear depressed centre and a surrounding erythematous ring. While familiar to those in farming communities, the presence of the nodule may be concerning to others, particularly the immunosuppressed. Milker's nodules are self-limited in immunocompetent individuals and heal without scarring within 8 weeks. Another member of the Parapoxvirus genus, the orf virus, is also transmitted from animals to humans by direct contact. While complications are rare, haematopoietic stem cell transplant recipients are at risk of graft-versus-host disease, as the parapoxvirus may trigger these complications in immunocompromised individuals. In addition, paravaccinia may serve as the antigen source for the development of erythema multiforme. The unique structure and replication process of viruses in the Poxvirus family, while includes the Parapoxvirus genus, have been a focus for treatment of infections and cancer. Manipulation of these viruses has demonstrated promising therapeutic possibilities as vectors for vaccines and oncologic therapy.


Subject(s)
Immunocompromised Host , Occupational Diseases/pathology , Poxviridae Infections/transmission , Aminoquinolines/therapeutic use , Animals , Antiviral Agents/therapeutic use , Diagnosis, Differential , Humans , Idoxuridine/therapeutic use , Imiquimod , Immunocompetence , Occupational Diseases/diagnosis , Occupational Diseases/drug therapy , Poxviridae Infections/diagnosis , Poxviridae Infections/drug therapy , Poxviridae Infections/pathology , Zoonoses
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