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Chang VY, He Y, Grohe S, Brady MR, Chan A, Kadam RS, Fang T, Pang A, Pohl K, Tran E, Li M, Kan J, Zhang Y, Lu JJ, Sasine JP, Himburg HA, Yue P, Chute JP. Epidermal growth factor augments the self-renewal capacity of aged hematopoietic stem cells. iScience 2024; 27:110306. [PMID: 39055915 PMCID: PMC11269946 DOI: 10.1016/j.isci.2024.110306] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/21/2023] [Revised: 02/19/2024] [Accepted: 06/17/2024] [Indexed: 07/28/2024] Open
Abstract
Hematopoietic aging is associated with decreased hematopoietic stem cell (HSC) self-renewal capacity and myeloid skewing. We report that culture of bone marrow (BM) HSCs from aged mice with epidermal growth factor (EGF) suppressed myeloid skewing, increased multipotent colony formation, and increased HSC repopulation in primary and secondary transplantation assays. Mice transplanted with aged, EGF-treated HSCs displayed increased donor cell engraftment within BM HSCs and systemic administration of EGF to aged mice increased HSC self-renewal capacity in primary and secondary transplantation assays. Expression of a dominant negative EGFR in Scl/Tal1+ hematopoietic cells caused increased myeloid skewing and depletion of long term-HSCs in 15-month-old mice. EGF treatment decreased DNA damage in aged HSCs and shifted the transcriptome of aged HSCs from genes regulating cell death to genes involved in HSC self-renewal and DNA repair but had no effect on HSC senescence. These data suggest that EGFR signaling regulates the repopulating capacity of aged HSCs.
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Affiliation(s)
- Vivian Y. Chang
- Division of Hematology-Oncology, Department of Pediatrics, UCLA, Los Angeles, CA, USA
- Children’s Discovery and Innovation Institute, UCLA, Los Angeles, CA, USA
- Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA, USA
| | - Yuwei He
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Samantha Grohe
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Morgan R. Brady
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Aldi Chan
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Rucha S. Kadam
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Tiancheng Fang
- Department of Molecular and Medical Pharmacology, UCLA, Los Angeles, CA, USA
| | - Amara Pang
- Division of Hematology/Oncology, Department of Medicine, University of California, Los Angeles, Los Angeles, CA, USA
| | - Katherine Pohl
- Division of Hematology/Oncology, Department of Medicine, University of California, Los Angeles, Los Angeles, CA, USA
| | - Evelyn Tran
- Division of Hematology/Oncology, Department of Medicine, University of California, Los Angeles, Los Angeles, CA, USA
| | - Michelle Li
- Division of Hematology/Oncology, Department of Medicine, University of California, Los Angeles, Los Angeles, CA, USA
| | - Jenny Kan
- Division of Hematology/Oncology, Department of Medicine, University of California, Los Angeles, Los Angeles, CA, USA
| | - Yurun Zhang
- Molecular Biology Institute, UCLA, Los Angeles, CA 90095, USA
| | - Josie J. Lu
- Applied Genomics, Computation and Translational Core, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Joshua P. Sasine
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - Heather A. Himburg
- Department of Radiation Oncology, Medical College of Wisconsin, Milwaukee, WI, USA
| | - Peibin Yue
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
| | - John P. Chute
- Division of Hematology & Cellular Therapy, Cedars Sinai Medical Center, Los Angeles, CA, USA
- Board of Governors Regenerative Medicine Institute, Cedars Sinai Medical Center, Los Angeles, CA 90095, USA
- Samuel Oschin Cancer Center, Cedars Sinai Medical Center, Los Angeles, CA 90095, USA
- Department of Medicine, Cedars Sinai Medical Center, Los Angeles, CA 91361, USA
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Sharma N, Ajima MNO, Rather MA, Sharma R, Ahmad I. Behavioural changes, DNA damage and histological alterations in Labeo rohita fingerlings in response to organic-coated silver nanoparticles. ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH INTERNATIONAL 2024; 31:47789-47800. [PMID: 39007970 DOI: 10.1007/s11356-024-34360-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/13/2023] [Accepted: 07/08/2024] [Indexed: 07/16/2024]
Abstract
Silver nanoparticles (AgNPs) have garnered significant global attention from researchers due to their unique physicochemical properties and wide-ranging applications in industry and medicine. However, their release into aquatic ecosystems has raised concerns regarding potential ecotoxicological consequences. The present study investigated the effects of polyvinyl pyrrolidone-coated silver nanoparticles on Labeo rohita fingerlings, focusing on behavioural reactions, genotoxic effects, histological changes and bioaccumulation. L. rohita fingerlings were exposed to polyvinyl pyrrolidone-coated silver nanoparticles with sizes ranging from 18 to 29 nm for 7 days at concentrations of 100, 200, 400 and 800 ug/l. The nanoparticle zeta potential was found to be extremely negative, measuring - 55.5 mV for 18 nm and - 31.4 mV for 29 nm. Behavioural abnormalities, including respiratory distress, reduced responsiveness and erratic swimming, were observed in exposed groups compared to controls, with severity increasing with higher nanoparticle concentrations. Genotoxicity assessment revealed significantly higher DNA damage in kidney cells compared to gill cells. Histological examination of gill tissues showed clogging in primary and secondary lamellae, along with distorted anatomy, necrosis and vacuolar atrophy in peripheral tubules of the kidneys. The kidneys exhibited greater nanoparticle accumulation than the gills with prolonged exposure. Moreover, 18 nm AgNPs induced more pronounced DNA damage and histological alterations in the kidney and gill tissues compared to 29 nm nanoparticles. This study elucidates the critical role of monitoring AgNPs in aquatic systems, providing essential data on their behaviour and environmental impacts. The findings highlight the need for improved detection techniques and effective management of AgNP contamination. Future research should focus on developing more sensitive analytical methods, understanding long-term ecological effects and exploring innovative remediation strategies.
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Affiliation(s)
- Niti Sharma
- Central Inland Fisheries Research Institute, Regional Centre, Guwahati, Assam, 781006, India
| | - Malachy N O Ajima
- Department of Fisheries and Aquaculture Technology, Federal University of Technology, P.M.B. 1526, Owerri, Nigeria
| | - Mohd Ashraf Rather
- Division of Fish Genetics and Biotechnology, Faculty of Fisheries Rangil Ganderbal, SKUAST-Kashmir, Srinagar, India.
| | - Rupam Sharma
- Fish Genetics and Biotechnology Division, Central Institute of Fisheries Education, Panch Marg Off Yari Road, Versova, Andheri West, Mumbai, India
| | - Ishtiyaq Ahmad
- Division of Fish Genetics and Biotechnology, Faculty of Fisheries Rangil Ganderbal, SKUAST-Kashmir, Srinagar, India
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Jaber N, Billet S. How to use an in vitro approach to characterize the toxicity of airborne compounds. Toxicol In Vitro 2024; 94:105718. [PMID: 37871865 DOI: 10.1016/j.tiv.2023.105718] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/22/2023] [Revised: 09/13/2023] [Accepted: 09/21/2023] [Indexed: 10/25/2023]
Abstract
As part of the development of new approach methodologies (NAMs), numerous in vitro methods are being developed to characterize the potential toxicity of inhalable xenobiotics (gases, volatile organic compounds, polycyclic aromatic hydrocarbons, particulate matter, nanoparticles). However, the materials and methods employed are extremely diverse, and no single method is currently in use. Method standardization and validation would raise trust in the results and enable them to be compared. This four-part review lists and compares biological models and exposure methodologies before describing measurable biomarkers of exposure or effect. The first section emphasizes the importance of developing alternative methods to reduce, if not replace, animal testing (3R principle). The biological models presented are mostly to cultures of epithelial cells from the respiratory system, as the lungs are the first organ to come into contact with air pollutants. Monocultures or cocultures of primary cells or cell lines, as well as 3D organotypic cultures such as organoids, spheroids and reconstituted tissues, but also the organ(s) model on a chip are examples. The exposure methods for these biological models applicable to airborne compounds are submerged, intermittent, continuous either static or dynamic. Finally, within the restrictions of these models (i.e. relative tiny quantities, adhering cells), the mechanisms of toxicity and the phenotypic markers most commonly examined in models exposed at the air-liquid interface (ALI) are outlined.
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Affiliation(s)
- Nour Jaber
- UR4492, Unité de Chimie Environnementale et Interactions sur le Vivant, Université du Littoral Côte d'Opale, Dunkerque, France
| | - Sylvain Billet
- UR4492, Unité de Chimie Environnementale et Interactions sur le Vivant, Université du Littoral Côte d'Opale, Dunkerque, France.
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Wu B, Zhang Y, You Y, Liang Y. Genotoxicity of chlorinated hydrophobic organic compounds extracted from a source of drinking water. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 2023; 267:115598. [PMID: 39492175 DOI: 10.1016/j.ecoenv.2023.115598] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/07/2023] [Revised: 10/10/2023] [Accepted: 10/12/2023] [Indexed: 11/05/2024]
Abstract
As precursors of disinfection by-products (DBPs), sediment hydrophobic organic compounds (HPOCs) derived from six reservoirs along the Dongjiang River were chlorinated (Cl-WS) and then fractionated into two polar components (hydrophobic organic compounds (Cl-HOCs) and hydrophilic organic compounds (Cl-HICs)) by solid phase extraction. Based on Caco-2 cell exposure study, the genotoxicity of Cl-WS, Cl-HOCs and Cl-HICs was analyzed by reactive oxygen species (ROS) analysis, comet assay and ethoxyresorufin-O-deethylase assay. Protective effects of antioxidants (catalase, vitamin C and epigallocatechin 3-gallate (EGCG)) on genotoxicity of the chlorinated samples were investigated. The results showed that Cl-WS and its two fractions (Cl-HOCs and Cl-HICs) induced ROS, DNA damage and dioxin-like toxicity (TEQbio), and Cl-HICs were a major contributor to oxidative damage and TEQbio compared with Cl-HOCs. Antioxidants significantly reduced Cl-HOCs- and Cl-HICs-induced ROS, but had insignificant effect on DNA oxidative damage and TEQbio. In addition, EGCG showed higher efficacy in reducing DNA damage induced by Cl-HICs than by Cl-HOCs. To our knowledge, this is the first study to investigate the genotoxicity of Cl-WS and its two polar components in Caco-2 cells, as well as the protective effects of antioxidants on Cl-HOCs and Cl-HICs-induced ROS, DNA damage, and TEQbio. This study provides important toxicity information for water treatment industries in differentiating DBPs of different polarity.
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Affiliation(s)
- Binbin Wu
- Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
| | - Yanling Zhang
- Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
| | - Yuehua You
- Department of Stomatology, Longhua People's Hospital Afliated to Southern Medical University, Shenzhen 518109, Guangdong, China; School of Stomatology, Southern Medical University, Guangzhou 510515, Guangdong, China; Key Laboratory of Oral Microbiology and Medical Transformation of Shenzhen Longhua District, China.
| | - Yan Liang
- School of Resources and Environment, University of Electronic Science and Technology of China, Chengdu 611731, China; Shenzhen Institute for Advanced Study, University of Electronic Science and Technology of China, Shenzhen 518000, China.
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Kluska M, Piastowska-Ciesielska AW, Tokarz P. Cell Cycle Status Influences Resistance to Apoptosis Induced by Oxidative Stress in Human Breast Cancer Cells, Which Is Accompanied by Modulation of Autophagy. Curr Issues Mol Biol 2023; 45:6325-6338. [PMID: 37623218 PMCID: PMC10453102 DOI: 10.3390/cimb45080399] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2023] [Revised: 07/21/2023] [Accepted: 07/28/2023] [Indexed: 08/26/2023] Open
Abstract
Cancer cells are characterised by uncontrolled cell proliferation; however, some of them can temporarily arrest their cell cycle at the G0 or G1 phase, which could contribute to tumour heterogeneity and drug resistance. The cell cycle status plays a critical role in chemosensitivity; however, the influence of G0- and G1-arrest has not been elucidated. To study the cell cycle arrest-mediated resistance, we used MCF-7 cells and generated three populations of cells: (1) cells arrested in the G0-like phase, (2) cells that resumed the cell cycle after the G0-like phase and (3) cells arrested in early G1 with a history of G0-like arrest. We observed that both the G0-like- and the G1-arrested cells acquired resistance to apoptosis induced by oxidative stress, accompanied by a decreased intracellular reactive oxygen species and DNA damage. This effect was associated with increased autophagy, likely facilitating their survival at DNA damage insult. The cell cycle reinitiation restored a sensitivity to oxidative stress typical for cells with a non-modulated cell cycle, with a concomitant decrease in autophagy. Our results support the need for further research on the resistance of G0- and G1-arrested cancer cells to DNA-damaging agents and present autophagy as a candidate for targeting in anticancer treatment.
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Affiliation(s)
- Magdalena Kluska
- Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
| | | | - Paulina Tokarz
- Department of Molecular Genetics, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
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Velarde JMC, Bastos NM, Carneiro-Leite L, Borges LP, Vieira EG, Veríssimo-Silveira R, Ninhaus-Silveira A. Dimethyl acetamide and dimethyl sulfoxide associated at glucose and egg yolk for cryopreservation of Pseudoplatystoma corruscans semen. NEOTROPICAL ICHTHYOLOGY 2023. [DOI: 10.1590/1982-0224-2022-0071] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/22/2023]
Abstract
Abstract This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.
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Sykuła A, Nowak A, Garribba E, Dzeikala A, Rowińska-Żyrek M, Czerwińska J, Maniukiewicz W, Łodyga-Chruścińska E. Spectroscopic Characterization and Biological Activity of Hesperetin Schiff Bases and Their Cu(II) Complexes. Int J Mol Sci 2023; 24:ijms24010761. [PMID: 36614204 PMCID: PMC9821237 DOI: 10.3390/ijms24010761] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/29/2022] [Revised: 12/19/2022] [Accepted: 12/29/2022] [Indexed: 01/03/2023] Open
Abstract
The three Schiff base ligands, derivatives of hesperetin, HHSB (N-[2,3-dihydro-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chromen-4-ylidene]isonicotinohydrazide), HIN (N-[2,3-dihydro-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chromen-4-ylidene]benzhydrazide) and HTSC (N-[2,3-dihydro-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chromen-4-ylidene]thiosemicarbazide) and their copper complexes, CuHHSB, CuHIN, and CuHTSC were designed, synthesized and analyzed in terms of their spectral characterization and the genotoxic activity. Their structures were established using several methods: elemental analysis, FT-IR, UV-Vis, EPR, and ESI-MS. Spectral data showed that in the acetate complexes the tested Schiff bases act as neutral tridentate ligand coordinating to the copper ion through two oxygen (or oxygen and sulphur) donor atoms and a nitrogen donor atom. EPR measurements indicate that in solution the complexes keep their structures with the ligands remaining bound to copper(II) in a tridentate fashion with (O-, N, Oket) or (O-, N, S) donor set. The genotoxic activity of the compounds was tested against model tumour (HeLa and Caco-2) and normal (LLC-PK1) cell lines. In HeLa cells the genotoxicity for all tested compounds was noticed, for HHSB and CuHHSB was the highest, for HTSC and CuHTSC-the lowest. Generally, Cu complexes displayed lower genotoxicity to HeLa cells than ligands. In the case of Caco-2 cell line HHSB and HTSC induced the strongest breaks to DNA. On the other side, CuHHSB and CuHTSC induced the highest DNA damage against LLC-PK1.
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Affiliation(s)
- Anna Sykuła
- Faculty of Biotechnology and Food Sciences, Institute of Natural Products and Cosmetics, Lodz University of Technology, Stefanowskiego 2/22, 90-537 Lodz, Poland
| | - Adriana Nowak
- Department of Environmental Biotechnology, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, Wólczańska 171/173, 90-530 Lodz, Poland
| | - Eugenio Garribba
- Department of Medicine, Surgery and Pharmacy, University of Sassari, Viale San Pietro, I-07100 Sassari, Italy
| | - Aliaksandr Dzeikala
- Faculty of Biotechnology and Food Sciences, Institute of Natural Products and Cosmetics, Lodz University of Technology, Stefanowskiego 2/22, 90-537 Lodz, Poland
| | | | - Justyna Czerwińska
- Department of Occupational Safety Engineering, Faculty of Process and Environmental Engineering, Lodz University of Technology, Wólczańska 213, 90-924 Lodz, Poland
| | - Waldemar Maniukiewicz
- Faculty of Chemistry, Institute of General and Ecological Chemistry, Lodz University of Technology, Żeromskiego 116, 90-924 Lodz, Poland
| | - Elżbieta Łodyga-Chruścińska
- Faculty of Biotechnology and Food Sciences, Institute of Natural Products and Cosmetics, Lodz University of Technology, Stefanowskiego 2/22, 90-537 Lodz, Poland
- Correspondence:
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Sitarek P, Kowalczyk T, Synowiec E, Merecz-Sadowska A, Bangay G, Princiotto S, Śliwiński T, Rijo P. An Evaluation of the Novel Biological Properties of Diterpenes Isolated from Plectranthus ornatus Codd. In Vitro and In Silico. Cells 2022; 11:cells11203243. [PMID: 36291112 PMCID: PMC9600095 DOI: 10.3390/cells11203243] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/29/2022] [Revised: 10/09/2022] [Accepted: 10/10/2022] [Indexed: 11/16/2022] Open
Abstract
Plectranthus ornatus Codd, the genus Plectranthus of the Lamiaceae family, has been used as traditional medicine in Africa, India and Australia. Pharmacological studies show the use of this plant to treat digestive problems. In turn, leaves were used for their antibiotic properties in some regions of Brazil to treat skin infections. The present study examines the anti-inflammatory, antioxidant and cytotoxic effects of the halimane and labdane diterpenes (11R*,13E)-11-acetoxyhalima-5,13-dien-15-oic acid (HAL) and 1α,6β-diacetoxy-8α,13R*-epoxy-14-labden-11-one (PLEC) and the forskolin-like 1:1 mixture of 1,6-di-O-acetylforskolin and 1,6-di-O-acetyl-9-deoxyforskolin (MRC) isolated from P. ornatus on lung (A549) and leukemia (CCRF-CEM) cancer cell lines, and on normal human retinal pigment epithelial (ARPE-19) cell line in vitro. Additionally, molecular docking and computational approaches were used. ADMET properties were analysed through SwissADME and proTox-II—Prediction. The results indicate that all tested compounds significantly reduced the viability of the cancer cells and demonstrated no cytotoxic effects against the non-neoplastic cell line. The apoptosis indicators showed increased ROS levels for both the tested A549 and CCRF-CEM cancer cell lines after treatment. Furthermore, computational studies found HAL to exhibit moderate antioxidant activity. In addition, selected compounds changed mitochondrial membrane potential (MMP), and increased DNA damage and mitochondrial copy number for the CCRF-CEM cancer cell line; they also demonstrated anti-inflammatory effects on the ARPE-19 normal cell line upon lipopolysaccharide (LPS) treatment, which was associated with the modulation of IL-6, IL-8, TNF-α and GM-CSF genes expression. Docking studies gave indication about the lowest binding energy for 1,6-di-O-acetylforskolin docked into IL-6, TNF-α and GM-CSF, and 1,6-di-O-acetyl-9-deoxyforskolin docked into IL-8. The ADMET studies showed drug-likeness properties for the studied compounds. Thus, halimane and labdane diterpenes isolated from P. ornatus appear to offer biological potential; however, further research is necessary to understand their interactions and beneficial properties.
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Affiliation(s)
- Przemysław Sitarek
- Department of Biology and Pharmaceutical Botany, Medical University of Lodz, ul. Muszyńskiego 1, 90-151 Lodz, Poland
- Correspondence: (P.S.); (P.R.)
| | - Tomasz Kowalczyk
- Department of Molecular Biotechnology and Genetics, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland
| | - Ewelina Synowiec
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
| | - Anna Merecz-Sadowska
- Department of Computer Science in Economics, University of Lodz, 90-214 Lodz, Poland
| | - Gabrielle Bangay
- CBIOS—Lusófona University’s Research Center for Biosciences and Health Technologies, 1749-024 Lisbon, Portugal
| | - Salvatore Princiotto
- CBIOS—Lusófona University’s Research Center for Biosciences and Health Technologies, 1749-024 Lisbon, Portugal
| | - Tomasz Śliwiński
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
| | - Patricia Rijo
- Department of Molecular Biotechnology and Genetics, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland
- CBIOS—Lusófona University’s Research Center for Biosciences and Health Technologies, 1749-024 Lisbon, Portugal
- Instituto de Investigação do Medicamento (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, 1649-003 Lisbon, Portugal
- Correspondence: (P.S.); (P.R.)
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Arpini AP, De Lorenzo A, Moritz A, Pereira JP, Dias GM. Evaluation of DNA damage induced by ionizing radiation from myocardial perfusion imaging: a pilot study. BMC Cardiovasc Disord 2022; 22:394. [PMID: 36057570 PMCID: PMC9441099 DOI: 10.1186/s12872-022-02839-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/12/2022] [Accepted: 08/24/2022] [Indexed: 12/04/2022] Open
Abstract
Background As patient exposure to ionizing radiation raises concern about malignancy risks, this study evaluated the effect of ionizing radiation on patients undergoing myocardial perfusion imaging (MPI) using the comet assay, a method for detection of DNA damage. Methods Patients without cancer, acute or autoimmune diseases, recent surgery or trauma, were studied. Gated single-photon myocardial perfusion imaging was performed with Tc-99m sestamibi. Peripheral blood was collected before radiotracer injection at rest and 60–90 min after injection. Single-cell gel electrophoresis (comet assay) was performed with blood lymphocytes to detect strand breaks, which determine a “comet tail” of variable size, visually scored by 3 observers in a fluorescence microscope after staining (0: no damage, no tail; 1: small damage; 2: large damage; 3: full damage). A damage index was calculated as a weighted average of the cell scores. Results Among the 29 individuals included in the analysis, age was 65.3 ± 9.9 years and 18 (62.1%) were male. The injected radiotracer dose was 880.6 ± 229.4 MBq. Most cells (approximately 70%) remained without DNA fragmentation (class 0) after tracer injection. There were nonsignificant increases of classes 1 and 2 of damage. Class 3 was the least frequent both before and after radiotracer injection, but displayed a significant, 44% increase after injection. Conclusion While lymphocytes mostly remained in class 0, an increase in class 3 DNA damage was detected. This may suggest that, despite a probable lack of biologically relevant DNA damage, there is still a need for tracer dose reductions in MPI.
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Affiliation(s)
- Anna Paula Arpini
- Coordenação de Ensino E Pesquisa, Instituto Nacional de Cardiologia, Rua das Laranjeiras 374, Rio de Janeiro, RJ, Brazil
| | - Andrea De Lorenzo
- Coordenação de Ensino E Pesquisa, Instituto Nacional de Cardiologia, Rua das Laranjeiras 374, Rio de Janeiro, RJ, Brazil.
| | - Aniele Moritz
- Coordenação de Ensino E Pesquisa, Instituto Nacional de Cardiologia, Rua das Laranjeiras 374, Rio de Janeiro, RJ, Brazil
| | - Julia Passarelli Pereira
- Coordenação de Ensino E Pesquisa, Instituto Nacional de Cardiologia, Rua das Laranjeiras 374, Rio de Janeiro, RJ, Brazil
| | - Glauber Monteiro Dias
- Coordenação de Ensino E Pesquisa, Instituto Nacional de Cardiologia, Rua das Laranjeiras 374, Rio de Janeiro, RJ, Brazil.,Laboratório de Biologia Celular E Tecidual, Centro de Biociências E Biotecnologia, Universidade Estadual Do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, RJ, Brazil
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Gopi N, Iswarya A, Vijayakumar S, Jayanthi S, Nor SAM, Velusamy P, Vaseeharan B. Protective effects of dietary supplementation of probiotic Bacillus licheniformis Dahb1 against ammonia induced immunotoxicity and oxidative stress in Oreochromis mossambicus. Comp Biochem Physiol C Toxicol Pharmacol 2022; 259:109379. [PMID: 35609808 DOI: 10.1016/j.cbpc.2022.109379] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/19/2021] [Revised: 04/25/2022] [Accepted: 05/17/2022] [Indexed: 12/23/2022]
Abstract
The goal of this study was to assess the efficacy of probiotics in mitigating ammonia-induced toxicity in fish. Fish were divided into four groups: control, only probiotic, only ammonia, and combined ammonia + probiotic. For 8 weeks, the Oreochromis mossambicus were exposed to waterborne ammonia at 1.0 mg L-1 and/or dietary Bacillus licheniformis Dahb1 at 107 cfu g-1. After the 4th and 8th weeks, the fish were evaluated for growth performance, enzymatic and non-enzymatic antioxidant activities (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) reduced glutathione (GSH), neurotoxicity (acetylcholinesterase - AChE), non-specific immune responses (lysozyme (LYZ), myeloperoxidase (MPO), reactive nitrogen and oxygen species (RNS and ROS) and oxidative stress effects (lipid peroxidation (LPO), DNA damage)). Our results showed that in the absence of waterborne ammonia exposure, B. licheniformis Dahb1 significantly improved growth performance, enzymatic and non-enzymatic antioxidant capacity, AChE activity, non-specific immune response and decreased oxidative stress effect. Ammonia exposure resulted in significantly lower growth performance, reduced enzymatic and non-enzymatic antioxidant ability, decreased AChE activity, decreased non-specific immune response and increased oxidative stress effect. When O. mossambicus were exposed to ammonia, supplementation with B. licheniformis Dahb1 in the diet significantly increased survival, indicating that it may have a significant protective effect against ammonia toxicity by enhancing enzymatic and non-enzymatic antioxidant ability, activity of AChE, non-specific immune response and reduced oxidative stress effect. According to our findings, diet supplementation of B. licheniformis Dahb1 (107 cfu g-1) has the potential to combat ammonia toxicity in O. mossambicus.
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Affiliation(s)
- Narayanan Gopi
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Science Campus 6(th) Floor, Alagappa University, Karaikudi 630004,Tamil Nadu, India
| | - Arokiadhas Iswarya
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Science Campus 6(th) Floor, Alagappa University, Karaikudi 630004,Tamil Nadu, India; Department of Poultry and Aquaculture, Agricultural Research Organization, 7528809 Rishon, Letziyon, Israel
| | - Sekar Vijayakumar
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Science Campus 6(th) Floor, Alagappa University, Karaikudi 630004,Tamil Nadu, India; Marine College, Shandong University, Weihai 264209, PR China
| | - Sangily Jayanthi
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Science Campus 6(th) Floor, Alagappa University, Karaikudi 630004,Tamil Nadu, India
| | - Siti Azizah Mohd Nor
- Institute of Marine Biotechnology, Universiti Malaysia Terengganu, 21030 Kuala Terengganu, Terengganu, Malaysia
| | - Palaniyandi Velusamy
- Research & Development Wing, Sree Balaji Medical College and Hospital (SBMCH)- Bharath Institute of Higher Education and Research (BIHER), Chennai 600 044, Tamil Nadu, India
| | - Baskaralingam Vaseeharan
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Science Campus 6(th) Floor, Alagappa University, Karaikudi 630004,Tamil Nadu, India.
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Valorization of the Photo-Protective Potential of the Phytochemically Standardized Olive ( Olea europaea L.) Leaf Extract in UVA-Irradiated Human Skin Fibroblasts. Molecules 2022; 27:molecules27165144. [PMID: 36014384 PMCID: PMC9415354 DOI: 10.3390/molecules27165144] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/08/2022] [Revised: 08/03/2022] [Accepted: 08/09/2022] [Indexed: 11/17/2022] Open
Abstract
Leaves of Olea europaea are a by-product of the olive oil industry and a dietary supplement with acknowledged antioxidant and anti-inflammatory activity but underestimated photoprotective potential. We investigated the protective effects of the LC-PDA-MS/MS standardized ethanol-water extract of olive leaves (OLE), containing 26.2% total phenols and 22.2% oleuropein, with underlying mechanisms against the UVA-induced oxidative damage in human dermal fibroblasts. Hs68 cells were pre-treated (24 h) with OLE (2.5-25 μg/mL) or the reference antioxidants, quercetin and ascorbic acid (25 μg/mL), followed by irradiation (8 J/cm2). OLE significantly reduced the UVA-induced DNA damage and reactive oxygen species (ROS) overproduction and increased the thioredoxin reductase (TrxR) expression and post-radiation viability of fibroblasts by inhibiting their apoptosis. Both intrinsic and extrinsic apoptotic signaling pathways appeared to be inhibited by OLE, but the activity of caspase 9 was the most reduced. We hypothesized that the TrxR up-regulation by OLE could have prevented the UVA-induced apoptosis of Hs68 cells. In addition, a significant decrease in UVA-induced secretion levels of tumor necrosis factor (TNF-α) and interleukin-2 (IL-2) was shown in human lymphocyte culture in response to OLE treatment. In summary, our results support the beneficial effect of OLE in an in vitro model and indicate its great potential for use in the cosmetic and pharmaceutical industry as a topical photoprotective, antioxidant, and anti-inflammatory agent.
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Czubak-Prowizor K, Macieja A, Poplawski T, Zbikowska HM. Responses of human colon and breast adenocarcinoma cell lines (LoVo, MCF7) and non-tumorigenic mammary epithelial cells (MCF-10A) to the acellular fraction of packed red blood cells in the presence and absence of cisplatin. PLoS One 2022; 17:e0271193. [PMID: 35802725 PMCID: PMC9269965 DOI: 10.1371/journal.pone.0271193] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/24/2021] [Accepted: 06/24/2022] [Indexed: 11/19/2022] Open
Abstract
Perioperative blood transfusion in colorectal and some other cancer patients has been linked to the increased risk for recurrence, but a causal mechanism remains unclear. During the preparation and storage of packed red blood cells (PRBCs) bio-active substances accumulate in the acellular fraction (supernatant). Viability, proliferation, reactive oxygen species (ROS) levels, and DNA damage of colon (LoVo) and breast (MCF7) adenocarcinoma cells and non-tumorigenic MCF-10A cell line were determined in response to the supernatants of fresh and long-stored (day 42) PRBCs, leukoreduced (LR) or non-leukoreduced (NLR). The effect of supernatants on the cytotoxicity of cisplatin (cisPt) towards the cells was also examined. Supernatants, especially from a day 1 PRBCs, both LR and NLR, reduced the viability and inhibited proliferation of tumor cells (LoVo, MCF7), accompanying by the excessive ROS production, but these were not the case in MCF-10A. Moreover, supernatants had no effect on the cytotoxicity of cisPt against LoVo and MCF7 cells, while caused increased drug resistance in MCF-10A cells. The findings suggest the acellular fraction of PRBCs does not exhibit any pro-proliferative activity in the cancer cell lines studied. However, these are pioneering issues and require further research.
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Affiliation(s)
- Kamila Czubak-Prowizor
- Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
- Department of Cytobiology and Proteomics, Medical University of Lodz, Lodz, Poland
- * E-mail: ,
| | - Anna Macieja
- Department of Molecular Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Tomasz Poplawski
- Department of Chemistry and Clinical Biochemistry, Medical University of Lodz, Lodz, Poland
| | - Halina Malgorzata Zbikowska
- Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
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Czubak-Prowizor K, Macieja A, Poplawski T, Zbikowska HM. Packed Red Blood Cell Supernatants Do Not Promote Growth or Cisplatin Resistance of Myeloid Leukemia K-562 Cells. J Blood Med 2022; 13:121-131. [PMID: 35283654 PMCID: PMC8906863 DOI: 10.2147/jbm.s349965] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2021] [Accepted: 02/09/2022] [Indexed: 11/23/2022] Open
Affiliation(s)
- Kamila Czubak-Prowizor
- Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, 90-236, Poland
- Department of Cytobiology and Proteomics, Medical University of Lodz, Lodz, 92-215, Poland
- Correspondence: Kamila Czubak-Prowizor, Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, Lodz, 90-236, Poland, Tel +48 42 635 44 83, Email ;
| | - Anna Macieja
- Department of Molecular Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, 90-236, Poland
| | - Tomasz Poplawski
- Department of Chemistry and Clinical Biochemistry, Medical University of Lodz, Lodz, 90-136, Poland
| | - Halina Malgorzata Zbikowska
- Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, 90-236, Poland
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Álvarez-Ortiz P, Ascacio-Valdés J, Vera-Reyes I, Esparza-González C, Rodríguez-Herrera R, Salinas-Santander M, del Ángel-Martínez M, Morlett-Chávez A. Purshia plicata Triggers and Regulates Proteins Related to Apoptosis in HeLa Cancer Cells. PLANTS (BASEL, SWITZERLAND) 2021; 10:2559. [PMID: 34961030 PMCID: PMC8707402 DOI: 10.3390/plants10122559] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/11/2021] [Revised: 06/04/2021] [Accepted: 06/09/2021] [Indexed: 05/12/2023]
Abstract
Cervical cancer represents a public health problem, develops resistance to traditional therapies and cost-of-treatment is high. These disadvantages have led to the search for alternative bioactive-compound-based therapies. Said bioactive compounds include phenolic compounds, flavonoids, and tannins. The present study aimed to evaluate the therapeutic effect of a P. plicata extract on the HeLa cell line. Viability and apoptosis assays were run on the two cell lines treated with the extract. The peptides, up- and down-expressed in both cell lines, were identified by PDQuest analysis software and high-performance liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS). Our results show that a 500 mg/L treatment deregulated cell viability, with different apoptotic morphologies observed which are associated with the presence of bio-compounds, which up- and down-regulated the peptides. In conclusion, P. plicata regulates proteins associated with apoptosis in HeLa cancer cells.
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Affiliation(s)
- Patricia Álvarez-Ortiz
- Laboratory of Molecular Biology, Chemistry School, Autonomous University of Coahuila, Saltillo 25280, Mexico;
| | - Juan Ascacio-Valdés
- Bioprocesses and Bioproducts Research Group and Laboratory of Molecular Biology, Food Research Department, Chemistry School, Autonomous University of Coahuila, Saltillo 25280, Mexico; (J.A.-V.); (R.R.-H.)
| | - Ileana Vera-Reyes
- Proteomics Laboratory, Agricultural Plant Science and Biotechnology, Research Center for Applied Chemistry, Blvd. Enrique Reyna 140, Saltillo 25294, Mexico;
| | - Cecilia Esparza-González
- Laboratory of Histology, Dentistry School, Autonomous University of Coahuila, Saltillo 25125, Mexico;
| | - Raúl Rodríguez-Herrera
- Bioprocesses and Bioproducts Research Group and Laboratory of Molecular Biology, Food Research Department, Chemistry School, Autonomous University of Coahuila, Saltillo 25280, Mexico; (J.A.-V.); (R.R.-H.)
| | - Mauricio Salinas-Santander
- Laboratory of Molecular Biology, Health Research Department, Medicine School, Autonomous University of Coahuila, Saltillo 25000, Mexico; (M.S.-S.); (M.d.Á.-M.)
| | - Mayela del Ángel-Martínez
- Laboratory of Molecular Biology, Health Research Department, Medicine School, Autonomous University of Coahuila, Saltillo 25000, Mexico; (M.S.-S.); (M.d.Á.-M.)
| | - Antonio Morlett-Chávez
- Laboratory of Molecular Biology, Chemistry School, Autonomous University of Coahuila, Saltillo 25280, Mexico;
- Laboratory of Molecular Biology, Health Research Department, Medicine School, Autonomous University of Coahuila, Saltillo 25000, Mexico; (M.S.-S.); (M.d.Á.-M.)
- Clinical Laboratory Department, General Hospital No. 2, Mexican Institute of Social Security, Saltillo 25017, Mexico
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15
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Adel RM, Lotfy RA, Darwish AS, Amer AS. Destructive effect of iron overload in brain tissue of albino rats: Ameliorative role of silver immobilized organo-modified casein nanocomposite as co-treating agent with Deferasirox. J Trace Elem Med Biol 2021; 67:126794. [PMID: 34052583 DOI: 10.1016/j.jtemb.2021.126794] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/28/2020] [Revised: 05/03/2021] [Accepted: 05/17/2021] [Indexed: 01/19/2023]
Abstract
BACKGROUND Iron (Fe) is one of the most essential trace elements in the body that play crucial role in organisms' survival, however, excess deposition of it puts patients at higher risk of iron overload and tissue injury through production of reactive oxygen species (ROS), elevation of oxidative stress, development of endocrine disorders among which hypogonadism, and increased incidence of cells damage in vital organs. As deferasirox (DFX) is an approved Fe chelator drug, its inability to cross blood brain barrier (BBB) remains a definite obstacle against its use as Fe chelator in the brain. Lately, attention to nanoparticles usage in researches has been widely grown since their role in improving drug therapeutic effects and scavenging free radicals make them good candidates as chelating and antioxidant agents. AIMS Herein, after induction of iron overload, organo-modified casein immobilized silver nanocomposite (Ag@Tr-CA) was designed and explored as combined therapy with DFX drug to develop its penetrating efficiency toward BBB and its Fe chelating affinity. Moreover, to distinguish the advanced antioxidant character as well as the beneficial impact of it on lowering brain's oxidative stress. Meanwhile, its capability in regulating serum pituitary hormones such as follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and testosterone (T), ameliorating DNA damage, and improving brain's histopathological alterations was also assessed. METHODS The physicochemical characteristics of Ag@Tr-CA was carried out using X-ray powder diffractometry (XRD), Fourier transform infrared (FTIR), dynamic light scattering (DLS), field emission scanning electron microscope (FE-SEM), and high-resolution transmission electron microscope (HR-TEM) analyses. Effect of iron overload and subsequent treatment with DFX + Ag@Tr-CA on brain of adult male albino rats were evaluated using colorimetric methods to determine brain Fe concentration and brain oxidative stress biomarkers. Assessment of serum Fe indices and serum pituitary hormones (FSH, LH, PRL) and T were estimated by ELISA technique. Determination of DNA damage in cerebral cortex cells was accomplished using the alkaline version of comet assay, while detection of brain's histopathological alterations was performed by examination of H&E sections under light microscope. RESULTS The physicochemical characteristics of Ag@Tr-CA showing the proficiency of Ag nanoparticles (∼35 nm) in creating highly-ordered negatively charged micro-sized casein particles (∼450 μm). After induction of iron overload, DFX + Ag@Tr-CA combination efficiently down brain Fe concentration, brain oxidative stress markers, and DNA damage in cerebral cortex cells linked with improvements in brain histopathological alterations. Comparing DFX therapeutic action alone to its combination to whether Ag@Tr-CA or Tr-CA (organo-modified cross-linked casein nanoparticles) as co-treating agents revealed no significant effect on serum Fe indices, FSH, LH, PRL, and T against iron overload disease. CONCLUSION The present results showed that combination of Ag@Tr-CA nanocomposite with DFX makes it a promising co-treating agent against iron overload through improving the physiological, molecular, and histological structure of the brain in iron overloaded rats.
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Affiliation(s)
- Rana M Adel
- Zoology Department, Faculty of Women for Arts, Science and Education, Ain Shams University, 11757, Cairo, Egypt.
| | - Rania A Lotfy
- Zoology Department, Faculty of Women for Arts, Science and Education, Ain Shams University, 11757, Cairo, Egypt.
| | - Atef S Darwish
- Chemistry Department, Faculty of Science, Ain Shams University, 11566, Cairo, Egypt.
| | - Amany S Amer
- Zoology Department, Faculty of Women for Arts, Science and Education, Ain Shams University, 11757, Cairo, Egypt.
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Lyu L, Lin TC, McCarty N. TRIM44 mediated p62 deubiquitination enhances DNA damage repair by increasing nuclear FLNA and 53BP1 expression. Oncogene 2021; 40:5116-5130. [PMID: 34211088 PMCID: PMC9744239 DOI: 10.1038/s41388-021-01890-7] [Citation(s) in RCA: 16] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2020] [Revised: 05/20/2021] [Accepted: 06/04/2021] [Indexed: 12/14/2022]
Abstract
Cancer cells show increases in protein degradation pathways, including autophagy, during progression to meet the increased protein degradation demand and support cell survival. On the other hand, reduced autophagy activity during aging is associated with a reduced DNA damage response and increased genomic instability. Therefore, it is a puzzling how DNA repair can be increased in cancer cells that are resistant to chemotherapies or during progression when autophagy activity is intact or increased. We discovered that tripartite motif containing 44 (TRIM44) is a pivotal element regulating the DNA damage response in cancer cells with intact autophagy. TRIM44 deubiquitinates p62, an autophagy substrate, which leads to its oligomerization. This prevents p62 localization to the nucleus upon irradiation. Increased cytoplasmic retention of p62 by TRIM44 prevents the degradation of FLNA and 53BP1, which increases DNA damage repair. Together, our data support TRIM44 a potential therapeutic target for therapy-resistant tumor cells with intact autophagy.
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Affiliation(s)
- Lin Lyu
- Center for Stem Cell and Regenerative Disease, Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases (IMM), the University of Texas-Health Science Center at Houston, Houston, Texas, 77030, USA
| | - Tsung-Chin Lin
- Center for Stem Cell and Regenerative Disease, Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases (IMM), the University of Texas-Health Science Center at Houston, Houston, Texas, 77030, USA
| | - Nami McCarty
- Center for Stem Cell and Regenerative Disease, Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases (IMM), the University of Texas-Health Science Center at Houston, Houston, Texas, 77030, USA.,Correspondence: Nami McCarty, Ph.D., University of Texas-Health Science Center at Houston, 1825 Pressler St., IMM-630A, Houston, TX 77030, USA, , Tel: 713-500-2495, Fax: 713-500-2424
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17
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Kinetics of DNA Repair in Vicia faba Meristem Regeneration Following Replication Stress. Cells 2021; 10:cells10010088. [PMID: 33430297 PMCID: PMC7825715 DOI: 10.3390/cells10010088] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2020] [Revised: 12/29/2020] [Accepted: 01/04/2021] [Indexed: 12/14/2022] Open
Abstract
The astonishing survival abilities of Vicia faba, one the earliest domesticated plants, are associated, among other things, to the highly effective replication stress response system which ensures smooth cell division and proper preservation of genomic information. The most crucial pathway here seems to be the ataxia telangiectasia-mutated kinase (ATM)/ataxia telangiectasia and Rad3-related kinase (ATR)-dependent replication stress response mechanism, also present in humans. In this article, we attempted to take an in-depth look at the dynamics of regeneration from the effects of replication inhibition and cell cycle checkpoint overriding causing premature chromosome condensation (PCC) in terms of DNA damage repair and changes in replication dynamics. We were able to distinguish a unique behavior of replication factors at the very start of the regeneration process in the PCC-induced cells. We extended the experiment and decided to profile the changes in replication on the level of a single replication cluster of heterochromatin (both alone and with regard to its position in the nucleus), including the mathematical profiling of the size, activity and shape. The results obtained during these experiments led us to the conclusion that even “chaotic” events are dealt with in a proper degree of order.
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18
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Santos NCND, Soares BM, Pessoa CDÓ, Freitas LMAD, Cruz JPP, Ramos MESP, Pithon MM, Cerqueira EMDM. Evaluation of the Genotoxicity of Endodontic Materials for Deciduous Teeth Using the Comet Assay. PESQUISA BRASILEIRA EM ODONTOPEDIATRIA E CLÍNICA INTEGRADA 2021. [DOI: 10.1590/pboci.2021.062] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
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Light Stability, Pro-Apoptotic and Genotoxic Properties of Silver (I) Complexes of Metronidazole and 4-Hydroxymethylpyridine against Pancreatic Cancer Cells In Vitro. Cancers (Basel) 2020; 12:cancers12123848. [PMID: 33419296 PMCID: PMC7767315 DOI: 10.3390/cancers12123848] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2020] [Revised: 12/12/2020] [Accepted: 12/18/2020] [Indexed: 12/04/2022] Open
Abstract
Simple Summary Antimicrobial properties of silver (I) ion and its complexes with metronidazole and 4-hydroxymethylpyridine are well recognized. However, little is known about its anticancer activity toward human pancreatic cancer cells. Our in vitro study revealed that silver (I) ion and its complexes with metronidazole and 4-hydroxymethylpyridine induced pancreatic cancer cells death associated with genotoxic and proapoptotic properties. In turn, the stability of active substances is of crucial importance because it determines the efficacy and applicability in clinical use. Therefore, we also evaluated photostability of silver (I) nitrate and its complexes with metronidazole and 4- hydroxymethylpyridine. Our results showed that studied complexes are more photochemically stable than silver salts, which makes them better candidates for clinical therapy. Abstract Antimicrobial properties of silver (I) ion and its complexes are well recognized. However, recent studies suggest that both silver (I) ion and its complexes possess anticancer activity associated with oxidative stress-induced apoptosis of various cancer cells. In this study, we aimed to investigate whether silver nitrate and its complexes with metronidazole and 4-hydroxymethylpyridine exert anticancer action against human pancreatic cancer cell lines (PANC-1 and 1.2B4). In the study, we compared decomposition speed for silver complexes under the influence of daylight and UV-A (ultraviolet-A) rays. We employed the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazonium bromide) assay to evaluate the cytotoxicity and the alkaline comet assay to determine genotoxicity of silver nitrate and its complexes. Flow cytometry and the Annexin V-FITC/PI apoptosis detection kit were used to detect the apoptosis of human pancreatic cancer cells. We found a dose dependent decrease of both pancreatic cancer cell line viability after exposure to silver nitrate and its complexes. The flow cytometry analysis confirmed that cell death occurred mainly via apoptosis. We also documented that the studied compounds induced DNA damage. Metronidazole and 4-hydroxymethylpyridine alone did not significantly affect viability and level of DNA damage of pancreatic cancer cell lines. Complex compounds showed better stability than AgNO3, which decomposed slower than when exposed to light. UV-A significantly influences the speed of silver salt decomposition reaction. To conclude, obtained data demonstrated that silver nitrate and its complexes exerted anticancer action against human pancreatic cancer cells.
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20
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Marcelino JM, Villas Boas GR, Cunha M, Deus Júnior R, Castro LH, Araújo FH, Traesel GK, Dos Santos AC, Souza RI, Paes M, Gubert P, Guterres ZDR, de Lima FF, Silva T, Silva RC, Cardoso CAL, Argandoña EJ, Macorini LF, Oesterreich SA. Determination of preclinical safety of oil obtained from Pachira aquatica Aublet (Malvaceae) seeds: histopathological, biochemical, hematological, and genetic toxicity studies in rats. Drug Chem Toxicol 2020; 45:1504-1521. [PMID: 33252270 DOI: 10.1080/01480545.2020.1845713] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/03/2023]
Abstract
Pachira aquatica is a species used for medicinal and food purposes and has numerous phytochemicals that may have systemic toxic effects and damage to genetic material. This study aimed to evaluate acute and short-term oral toxicity, as well as genotoxic and clastogenic effects of oil extracted from P. aquatica (PASO) seeds in rats and Drosophila melanogaster. The results obtained with biochemical and hematological analyses did not show significant changes in any evaluated parameters when compared with reference values for the species used in the study. Data from the histopathological analysis corroborated results found in this study. These findings indicate low acute and short-term toxicity following oral PASO exposure in rats under the experimental conditions tested. Tests performed in rats showed that PASO did not present significant genotoxic or clastogenic effects on the cells analyzed with the three doses tested. Treatment with PASO in the offspring of HB crossing, which showed high cytochrome P450 levels, did not exhibit genotoxic activity, as demonstrated by the SMART test. These results suggest that products from the hepatic oil metabolism did not show genotoxicity under the conditions tested. Together, the results indicate that, under the experimental conditions tested, PASO is safe for repeated intake. As PASO exhibited low potential to cause harmful effects on living organisms, our study encourages further research aimed at assessing its pharmacological activity, since it is a widely consumed plant.
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Affiliation(s)
| | - Gustavo Roberto Villas Boas
- Research Group on Development of Pharmaceutical Products (P&DProFar), Center for Biological and Health Sciences, Federal University of Western Bahia, Bahia, Brazil
| | - Mila Cunha
- Research Group on Development of Pharmaceutical Products (P&DProFar), Center for Biological and Health Sciences, Federal University of Western Bahia, Bahia, Brazil
| | - Ramão Deus Júnior
- Faculty of Health Sciences, Federal University of Grande Dourados, Dourados, Brazil
| | - Luis Henrique Castro
- Faculty of Health Sciences, Federal University of Grande Dourados, Dourados, Brazil
| | | | | | | | | | - Marina Paes
- Research Group on Development of Pharmaceutical Products (P&DProFar), Center for Biological and Health Sciences, Federal University of Western Bahia, Bahia, Brazil
| | - Priscila Gubert
- Research Group on Development of Pharmaceutical Products (P&DProFar), Center for Biological and Health Sciences, Federal University of Western Bahia, Bahia, Brazil.,Laboratory of Imunopathology KeizoAsami, Department of Biochemistry, Federal University of Pernambuco, Recife, Brazil
| | - Zaira da Rosa Guterres
- Laboratory of Phytogenesis and Mutagenesis, State University of Mato Grosso do Sul, Mato Grosso do Sul, Brazil
| | | | - Tania Silva
- Postgraduate Program in Environmental Science and Technology, Federal University of Grande Dourados, Mato Grosso do Sul, Brazil
| | - Rogério César Silva
- Chemistry Course, State University of Mato Grosso do Sul, Mato Grosso do Sul, Brazil
| | | | - Eliana Janet Argandoña
- Faculty of Engineering - FAEN/UFGD, Federal University of Grande Dourados, Mato Grosso do Sul, Brazil
| | - Luis Fernando Macorini
- Faculty of Biological Sciences and Health, University Center of Grande Dourados, Mato Grosso do Sul, Brazil
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Perini V, Schacke M, Liddle P, Vilchez-Larrea S, Keszenman DJ, Lafon-Hughes L. PARP Inhibitor Olaparib Causes No Potentiation of the Bleomycin Effect in VERO Cells, Even in the Presence of Pooled ATM, DNA-PK, and LigIV Inhibitors. Int J Mol Sci 2020; 21:E8288. [PMID: 33167404 PMCID: PMC7663819 DOI: 10.3390/ijms21218288] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2020] [Revised: 10/14/2020] [Accepted: 10/19/2020] [Indexed: 11/25/2022] Open
Abstract
Poly(ADP-ribosyl)polymerase (PARP) synthesizes poly(ADP-ribose) (PAR), which is anchored to proteins. PAR facilitates multiprotein complexes' assembly. Nuclear PAR affects chromatin's structure and functions, including transcriptional regulation. In response to stress, particularly genotoxic stress, PARP activation facilitates DNA damage repair. The PARP inhibitor Olaparib (OLA) displays synthetic lethality with mutated homologous recombination proteins (BRCA-1/2), base excision repair proteins (XRCC1, Polβ), and canonical nonhomologous end joining (LigIV). However, the limits of synthetic lethality are not clear. On one hand, it is unknown whether any limiting factor of homologous recombination can be a synthetic PARP lethality partner. On the other hand, some BRCA-mutated patients are not responsive to OLA for still unknown reasons. In an effort to help delineate the boundaries of synthetic lethality, we have induced DNA damage in VERO cells with the radiomimetic chemotherapeutic agent bleomycin (BLEO). A VERO subpopulation was resistant to BLEO, BLEO + OLA, and BLEO + OLA + ATM inhibitor KU55933 + DNA-PK inhibitor KU-0060648 + LigIV inhibitor SCR7 pyrazine. Regarding the mechanism(s) behind the resistance and lack of synthetic lethality, some hypotheses have been discarded and alternative hypotheses are suggested.
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Affiliation(s)
- Valentina Perini
- Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Departamento de Genética, Montevideo 11.600, Uruguay; (V.P.); (M.S.); (P.L.)
| | - Michelle Schacke
- Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Departamento de Genética, Montevideo 11.600, Uruguay; (V.P.); (M.S.); (P.L.)
| | - Pablo Liddle
- Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Departamento de Genética, Montevideo 11.600, Uruguay; (V.P.); (M.S.); (P.L.)
| | - Salomé Vilchez-Larrea
- Instituto de Investigaciones en Ingeniería Genética y Biología Molecular “Dr. Héctor N. Torres”, Consejo Nacional de Investigaciones Científicas y Técnicas, Ciudad Autónoma de Buenos Aires 1428, Argentina;
| | - Deborah J. Keszenman
- Laboratorio de Radiobiología Médica y Ambiental, Grupo de Biofisicoquímica, Centro Universitario Regional Litoral Norte, Universidad de la República (UdelaR), Salto 50.000, Uruguay
| | - Laura Lafon-Hughes
- Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Departamento de Genética, Montevideo 11.600, Uruguay; (V.P.); (M.S.); (P.L.)
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22
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Szejk-Arendt M, Czubak-Prowizor K, Macieja A, Poplawski T, Olejnik AK, Pawlaczyk-Graja I, Gancarz R, Zbikowska HM. Polyphenolic-polysaccharide conjugates from medicinal plants of Rosaceae/Asteraceae family protect human lymphocytes but not myeloid leukemia K562 cells against radiation-induced death. Int J Biol Macromol 2020; 156:1445-1454. [DOI: 10.1016/j.ijbiomac.2019.11.186] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/22/2019] [Revised: 11/14/2019] [Accepted: 11/21/2019] [Indexed: 01/21/2023]
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Fang T, Zhang Y, Chang VY, Roos M, Termini CM, Signaevskaia L, Quarmyne M, Lin PK, Pang A, Kan J, Yan X, Javier A, Pohl K, Zhao L, Scott P, Himburg HA, Chute JP. Epidermal growth factor receptor-dependent DNA repair promotes murine and human hematopoietic regeneration. Blood 2020; 136:441-454. [PMID: 32369572 PMCID: PMC7378456 DOI: 10.1182/blood.2020005895] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2020] [Accepted: 04/13/2020] [Indexed: 02/07/2023] Open
Abstract
Chemotherapy and irradiation cause DNA damage to hematopoietic stem cells (HSCs), leading to HSC depletion and dysfunction and the risk of malignant transformation over time. Extrinsic regulation of HSC DNA repair is not well understood, and therapies to augment HSC DNA repair following myelosuppression remain undeveloped. We report that epidermal growth factor receptor (EGFR) regulates DNA repair in HSCs following irradiation via activation of the DNA-dependent protein kinase-catalytic subunit (DNA-PKcs) and nonhomologous end joining (NHEJ). We show that hematopoietic regeneration in vivo following total body irradiation is dependent upon EGFR-mediated repair of DNA damage via activation of DNA-PKcs. Conditional deletion of EGFR in hematopoietic stem and progenitor cells (HSPCs) significantly decreased DNA-PKcs activity following irradiation, causing increased HSC DNA damage and depressed HSC recovery over time. Systemic administration of epidermal growth factor (EGF) promoted HSC DNA repair and rapid hematologic recovery in chemotherapy-treated mice and had no effect on acute myeloid leukemia growth in vivo. Further, EGF treatment drove the recovery of human HSCs capable of multilineage in vivo repopulation following radiation injury. Whole-genome sequencing analysis revealed no increase in coding region mutations in HSPCs from EGF-treated mice, but increased intergenic copy number variant mutations were detected. These studies demonstrate that EGF promotes HSC DNA repair and hematopoietic regeneration in vivo via augmentation of NHEJ. EGF has therapeutic potential to promote human hematopoietic regeneration, and further studies are warranted to assess long-term hematopoietic effects.
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Affiliation(s)
| | | | - Vivian Y Chang
- Pediatric Hematology/Oncology
- Jonsson Comprehensive Cancer Center
| | - Martina Roos
- Jonsson Comprehensive Cancer Center
- Division of Hematology/Oncology, Department of Medicine
- Broad Stem Cell Research Center, and
| | | | | | | | - Paulina K Lin
- Division of Hematology/Oncology, Department of Medicine
| | - Amara Pang
- Division of Hematology/Oncology, Department of Medicine
| | - Jenny Kan
- Division of Hematology/Oncology, Department of Medicine
| | - Xiao Yan
- Department of Molecular and Medical Pharmacology
| | - Anna Javier
- Division of Hematology/Oncology, Department of Medicine
| | | | - Liman Zhao
- Division of Hematology/Oncology, Department of Medicine
| | - Peter Scott
- Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA
| | | | - John P Chute
- Jonsson Comprehensive Cancer Center
- Division of Hematology/Oncology, Department of Medicine
- Broad Stem Cell Research Center, and
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24
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Hanna DH, Saad GR. Nanocurcumin: preparation, characterization and cytotoxic effects towards human laryngeal cancer cells. RSC Adv 2020; 10:20724-20737. [PMID: 35517737 PMCID: PMC9054308 DOI: 10.1039/d0ra03719b] [Citation(s) in RCA: 40] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/25/2020] [Accepted: 05/11/2020] [Indexed: 11/21/2022] Open
Abstract
The aim of the present study was to prepare curcumin nanoparticles (nanocurcumin) by a sol-oil method to improve curcumin absorption and bioavailability, and to investigate the therapeutic effects of the prepared nanoparticles on the inhibition mechanisms towards human Hep-2 cancer cells. The nanoparticles were characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, X-ray diffraction, and zeta potential analysis. The prepared curcumin nanoparticles possessed a narrow particle size distribution with an average diameter of 28 nm. The inhibition effects on the growth of human Hep-2 cells were investigated using neutral red uptake and lactate dehydrogenase assays. The results indicated that the nanocurcumin has a selective effect in inhibiting Hep-2 cell growth in a dose- and time-dependent mode with the most effective IC50 value (17 ± 0.31 μg ml−1) obtained after 48 h of incubation without any cytotoxic effects on normal cells. This IC50 value of nanocurcumin revealed a significant increase of early and late apoptotic cells with an intense comet nucleus of Hep-2 cells as a marker of DNA damage. Flow cytometry analysis of the progression of apoptosis in nanocurcumin Hep-2 treated cells showed that arresting in the cell cycle in the G2/M phase with increasing apoptotic cells in the sub-G1 phase. At the same time, real-time PCR analysis indicated that the treatment of Hep-2 cells with nanocurcumin resulted in upregulation of P53, Bax, and Caspase-3, whereas there was downregulation of Bcl-XL. These findings gave insights into understanding that the inhibition mechanisms of nanocurcumin on the proliferation of Hep-2 cancer cells was through the G2/M cell cycle arrest and the induction of apoptosis was dependent on Caspase-3 and p53 activation. However, in vivo studies with an animal model are essential to validate these results. The aim of this study was to prepare curcumin nanoparticles using a sol–oil method to improve curcumin absorption and bioavailability, and to investigate the therapeutic effect of the prepared nanoparticles on the inhibition mechanisms toward human Hep-2 cancer cells.![]()
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Affiliation(s)
- Demiana H. Hanna
- Department of Chemistry
- Faculty of Science
- Cairo University
- Giza 12613
- Egypt
| | - Gamal R. Saad
- Department of Chemistry
- Faculty of Science
- Cairo University
- Giza 12613
- Egypt
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25
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Carneiro-Leite L, Bashiyo-Silva C, Oliveira YAA, Borges LP, Sanchez MP, Silva LGD, Lobato SIR, Rodrigues-Lisoni FC, Veríssimo-Silveira R, Ninhaus-Silveira A. Seminal characteristics and sensitivity of Astyanax lacustris (Characiformes: Characidae) sperm to cryoprotective solutions based on dimethylsufoxide and methylglicol. NEOTROPICAL ICHTHYOLOGY 2020. [DOI: 10.1590/1982-0224-2020-0039] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Abstract This study aimed to determine the semen characteristics of Astyanax lacustris after hormonal induction and to evaluate the sensitivity of the species sperm to cryoprotective solutions based on the cryoprotectants dimethyl sulfoxide and methyl glycol. Volume, color, sperm concentration, total motility and aspects of sperm movement were analyzed using “Integrated Semen Analysis System”. Three different extenders were tested: A) glucose 5%+egg yolk 10%, B) BTS®5% and C) glucose 5% and two permeable cryoprotectants: dimethyl sulfoxide (Me2SO) and methyl glycol (MTG). Fresh A. lacustris semen presented total motility of 76.6±11.2%, motility duration of 33.0±2.2s, sperm concentration of 7.22±3.2×109sptz/mL and seminal osmolality of 219±0.03mOsm/kg-1. The toxicity test showed the highest total motility values at the MTG15%+A, Me2SO15%+B and Me2SO10%+C dilutions, and the Me2SO10%+C and Me2SO15%+C dilutions presented the highest values for curvilinear velocity, linear velocity and average velocity. The tested protocol was not effective at maintaining the viability of A. lacustris semen after freezing because no motility was observed in any of the dilutions. However, the Comet Assay demonstrated that cryoprotectant solutions were effective in protecting the genetic material of cells, as DNA damage levels were low, with no difference between control and Me2SO10% + A, dilutions MTG10%+C, Me2SO10%+B and Me2SO15%+B.
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26
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Wu B, Zhang Y, Hong H, Hu M, Liu H, Chen X, Liang Y. Hydrophobic organic compounds in drinking water reservoirs: Toxic effects of chlorination and protective effects of dietary antioxidants against disinfection by-products. WATER RESEARCH 2019; 166:115041. [PMID: 31536888 DOI: 10.1016/j.watres.2019.115041] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/30/2019] [Revised: 08/28/2019] [Accepted: 09/02/2019] [Indexed: 06/10/2023]
Abstract
This study investigated formation and genotoxicities of disinfection by-products (DBPs) during chlorination of hydrophobic organic compounds (HOCs) extracted from six drinking water reservoirs around the Pearl River Delta region, China. Chemical analyses firstly showed that after chlorination aromatic HOCs (including polycyclic aromatic hydrocarbons, PAHs) decreased but DBPs (including chlorinated PAHs) increased, while genotoxicity assays revealed that the chlorination increased DNA damage in human Caco-2 cells. Although the link between DBPs and the genotoxicity was weak (insignificant correlations, p ≥ 0.05), we observed that chlorination of HOCs from more contaminated reservoirs in general resulted in higher genotoxicity. Secondly, remedial effects of catalase and dietary antioxidants (i.e. vitamin C and epigallocatechin gallate) in protecting cells against DBPs genotoxicity were detected. After 1 h treatment by the antioxidants, the DNA damage in Caco-2 cells (due to previous exposure to DBPs) significantly decreased (p < 0.05) in 7 out of a total of 18 treatments (38.9%). This is the first study demonstrating that catalase, vitamin C and epigallocatechin gallate protected human cells in vitro against DNA damage upon exposure to chlorinated genotoxic DBPs.
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Affiliation(s)
- Binbin Wu
- Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China; University of Chinese Academy of Sciences, Beijing, 100049, China
| | - Yanling Zhang
- Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China
| | - Huachang Hong
- College of Geography and Environmental Science, Zhejiang Normal University, Jinhua, 321004, China.
| | - Mei Hu
- Shandong Institute for Food and Drug Control, Shandong, 250101, China
| | - Hailong Liu
- School of Resources and Environment, University of Electronic Science and Technology of China, Chengdu, 611731, China
| | - Xi Chen
- Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China; University of Chinese Academy of Sciences, Beijing, 100049, China; School of Resources and Environment, University of Electronic Science and Technology of China, Chengdu, 611731, China
| | - Yan Liang
- College of Geography and Environmental Science, Zhejiang Normal University, Jinhua, 321004, China; School of Resources and Environment, University of Electronic Science and Technology of China, Chengdu, 611731, China.
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27
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Silencing Lysine-Specific Histone Demethylase 1 (LSD1) Causes Increased HP1-Positive Chromatin, Stimulation of DNA Repair Processes, and Dysregulation of Proliferation by Chk1 Phosphorylation in Human Endothelial Cells. Cells 2019; 8:cells8101212. [PMID: 31591366 PMCID: PMC6829479 DOI: 10.3390/cells8101212] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2019] [Revised: 09/21/2019] [Accepted: 09/30/2019] [Indexed: 12/13/2022] Open
Abstract
The methylation of histone lysine residues modifies chromatin conformation and regulates the expression of genes implicated in cell metabolism. Lysine-specific demethylase 1 (LSD1) is a flavin-dependent monoamine oxidase that can demethylate mono- and dimethylated histone lysines 4 and 9 (H3K4 and H3K9). The removal of methyl groups from the lysine residues of histone and non-histone proteins was found to be an important regulatory factor of cell proliferation. However, its role has not been fully elucidated. In this study, we assessed LSD1-mediated cell cycle progression using a human endothelial cell model. The short hairpin RNA knockdown of LSD1 inhibits the G2/M phase of cell cycle progression by checkpoint kinase 1 (Chk1) phosphorylation (S137). We observed elevated DNA damage, which was consistent with the increased detection of double-strand breaks as well as purines and pyrimidines oxidation, which accompanied the activation of ATR/ATRIP signaling by H2AXS139 phosphorylation. The irreversible pharmacological inhibition of LSD1 by 2-phenylcyclopropylamine (2-PCPA) inactivated its enzymatic activity, causing significant changes in heterochromatin and euchromatin conformation assessed by chromatin assembly factor 1 subunit A (CAF1A) and heterochromatin protein 1 isoform α and γ (HP1α/γ) immunofluorescence analysis. We conclude that the knockdown of LSD1 in endothelial cells leads to increased HP1-positive chromatin, the stimulation of DNA repair processes, and the dysregulation of proliferation machinery.
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28
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Vitamin D Supplementation Reduces Both Oxidative DNA Damage and Insulin Resistance in the Elderly with Metabolic Disorders. Int J Mol Sci 2019; 20:ijms20122891. [PMID: 31200560 PMCID: PMC6628266 DOI: 10.3390/ijms20122891] [Citation(s) in RCA: 51] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2019] [Revised: 05/31/2019] [Accepted: 06/10/2019] [Indexed: 02/07/2023] Open
Abstract
Background: Research evidence indicates that vitamin D deficiency is involved in the pathogenesis of insulin resistance (IR) and associated metabolic disorders including hyperglycemia and dyslipidemia. It also suggested that vitamin D deficiency is associated with elevated levels of oxidative stress and its complications. Therefore, the aim of our study was to determine the effect of vitamin D supplementation on DNA damage and metabolic parameters in vitamin D deficient individuals aged >45 with metabolic disorders. Material and Methods: Of 98 initially screened participants, 92 subjects deficient in vitamin D were included in the study. They were randomly assigned to the following group: with vitamin D supplementation (intervention group, n = 48) and without supplementation (comparative group, n = 44). The patients from both groups were divided into two subgroups according to the presence or absence of type 2 diabetes (T2DM). The intervention group was treated with 2000 International Unit (IU) cholecalciferol/day between October and March for three months. At baseline and after three-month supplementation vitamin D concentration (25-OH)D3 and endogenous and oxidative DNA damage were determined. In addition, fast plasma glucose (FPG), fasting insulin, HbA1c and lipid fraction (total cholesterol (TC), low-density lipoprotein cholesterol (LDL), high-density lipoprotein cholesterol (HDL), triglyceride (TG)), as well as anthropometric measurements (weight, height) were gathered. The following IR-related parameters were calculated Homeostatic Model Assesment – Insulin Resistance (HOMA-IR) and TG/HDL ratio. Results: Three-month vitamin D supplementation increased the mean vitamin D concentration to generally accepted physiological level independently of T2DM presence. Importantly, vitamin D exposure decreased the level of oxidative DNA damage in lymphocytes of patients of intervention group. Among studied metabolic parameters, vitamin D markedly increased HDL level, decreased HOMA-IR, TG/HDL ratio. Furthermore, we found that HbA1c percentage diminished about 0.5% in T2DM patients supplemented with vitamin D. Conclusion: The current study demonstrated that daily 2000I U intake of vitamin D for three months decreased the level of oxidative DNA damage, a marker of oxidative stress, independently on T2DM presence. Furthermore, vitamin D reduced metabolic parameters connected with IR and improved glucose and lipid metabolism. Therefore, our results support the assertion that vitamin D, by reducing oxidative stress and improving of metabolic profile, may decrease IR and related diseases.
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29
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Paul T, Shukla SP, Kumar K, Poojary N, Kumar S. Effect of temperature on triclosan toxicity in Pangasianodon hypophthalmus (Sauvage, 1878): Hematology, biochemistry and genotoxicity evaluation. THE SCIENCE OF THE TOTAL ENVIRONMENT 2019; 668:104-114. [PMID: 30852190 DOI: 10.1016/j.scitotenv.2019.02.443] [Citation(s) in RCA: 32] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/26/2018] [Revised: 02/24/2019] [Accepted: 02/27/2019] [Indexed: 06/09/2023]
Abstract
The rising level of triclosan (TCS) in aquatic environment is raising concerns and in this context, evaluation of toxicity towards aquatic organisms under varying environmental conditions, especially temperature, is a pre-requisite for a better understanding of the toxic effects on specific metabolic processes. In this report, the mechanistic physiological responses of fish towards varying concentration of TCS at graded temperature were evaluated. The static renewal acute test was performed, and 96 h median lethal concentration (LC50) of TCS for Pangasianodon hypophthalmus was estimated and the values were 848.33, 1181.94 and 1356.96 μg L-1 at 25, 30 and 35 °C respectively. The chronic study was performed for 30 days at 1/5th and 1/10th concentration of the estimated LC50 of TCS at 25, 30 and 35 °C respectively. The chronic effects resulted in significant decrease in total erythrocyte count (TEC), hemoglobin (Hb), packed cell volume (PCV), mean corpuscular hemoglobin (MCH) and mean cell volume (MCV), while a significant increase in total leukocyte count (TLC), mean corpuscular hemoglobin concentration (MCHC) and red cell distribution width (RDW) was observed in TCS exposed groups at 25-35 °C. Further, a significant increase in activity of transaminase enzymes, lactate dehydrogenase (LDH) and antioxidant enzymes (superoxide dismutase) (SOD) and catalase (CAT) except glutathione-S-transferase (GST) in liver and acetylcholinesterase (AChE) in brain of the TCS exposed fish was recorded in all the above temperature range. Severe damage of DNA in nucleus of blood and liver cells, and high micronuclei frequency (MNi) was noticed in TCS exposed groups at 25 °C. The report provides convincing evidence for the effect of temperature on TCS toxicity. The findings will help in gaining a better insight into the change in toxicity of TCS in a natural environment where diurnal variations in temperature may be crucial in determining the overall extent of toxicity.
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Affiliation(s)
- Tapas Paul
- Aquatic Environment and Health Management Division, ICAR-CIFE, Mumbai 400061, India
| | - S P Shukla
- Aquatic Environment and Health Management Division, ICAR-CIFE, Mumbai 400061, India
| | - Kundan Kumar
- Aquatic Environment and Health Management Division, ICAR-CIFE, Mumbai 400061, India
| | - Nalini Poojary
- Aquatic Environment and Health Management Division, ICAR-CIFE, Mumbai 400061, India
| | - Saurav Kumar
- Aquatic Environment and Health Management Division, ICAR-CIFE, Mumbai 400061, India.
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30
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Gajski G, Žegura B, Ladeira C, Novak M, Sramkova M, Pourrut B, Del Bo' C, Milić M, Gutzkow KB, Costa S, Dusinska M, Brunborg G, Collins A. The comet assay in animal models: From bugs to whales - (Part 2 Vertebrates). MUTATION RESEARCH-REVIEWS IN MUTATION RESEARCH 2019; 781:130-164. [PMID: 31416573 DOI: 10.1016/j.mrrev.2019.04.002] [Citation(s) in RCA: 49] [Impact Index Per Article: 8.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/19/2018] [Revised: 03/26/2019] [Accepted: 04/10/2019] [Indexed: 12/20/2022]
Abstract
The comet assay has become one of the methods of choice for the evaluation and measurement of DNA damage. It is sensitive, quick to perform and relatively affordable for the evaluation of DNA damage and repair at the level of individual cells. The comet assay can be applied to virtually any cell type derived from different organs and tissues. Even though the comet assay is predominantly used on human cells, the application of the assay for the evaluation of DNA damage in yeast, plant and animal cells is also quite high, especially in terms of biomonitoring. The present extensive overview on the usage of the comet assay in animal models will cover both terrestrial and water environments. The first part of the review was focused on studies describing the comet assay applied in invertebrates. The second part of the review, (Part 2) will discuss the application of the comet assay in vertebrates covering cyclostomata, fishes, amphibians, reptiles, birds and mammals, in addition to chordates that are regarded as a transitional form towards vertebrates. Besides numerous vertebrate species, the assay is also performed on a range of cells, which includes blood, liver, kidney, brain, gill, bone marrow and sperm cells. These cells are readily used for the evaluation of a wide spectrum of genotoxic agents both in vitro and in vivo. Moreover, the use of vertebrate models and their role in environmental biomonitoring will also be discussed as well as the comparison of the use of the comet assay in vertebrate and human models in line with ethical principles. Although the comet assay in vertebrates is most commonly used in laboratory animals such as mice, rats and lately zebrafish, this paper will only briefly review its use regarding laboratory animal models and rather give special emphasis to the increasing usage of the assay in domestic and wildlife animals as well as in various ecotoxicological studies.
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Affiliation(s)
- Goran Gajski
- Mutagenesis Unit, Institute for Medical Research and Occupational Health, Zagreb, Croatia.
| | - Bojana Žegura
- Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, Ljubljana, Slovenia
| | - Carina Ladeira
- H&TRC - Health & Technology Research Center, Escola Superior de Tecnologia da Saúde, Instituto Politécnico de Lisboa, Lisbon, Portugal; Centro de Investigação e Estudos em Saúde de Publica, Universidade Nova de Lisboa, Lisbon, Portugal
| | - Matjaž Novak
- Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, Ljubljana, Slovenia
| | - Monika Sramkova
- Biomedical Research Center, Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovakia
| | - Bertrand Pourrut
- EcoLab, Université de Toulouse, CNRS, INPT, UPS, Toulouse, France
| | - Cristian Del Bo'
- DeFENS-Division of Human Nutrition, University of Milan, Milan, Italy
| | - Mirta Milić
- Mutagenesis Unit, Institute for Medical Research and Occupational Health, Zagreb, Croatia
| | | | - Solange Costa
- Environmental Health Department, National Health Institute Dr. Ricardo Jorge, Porto, Portugal; EPIUnit - Instituto de Saúde Pública, Universidade do Porto, Porto, Portugal
| | - Maria Dusinska
- Health Effects Laboratory, Department of Environmental Chemistry-MILK, NILU - Norwegian Institute for Air Research, Kjeller, Norway
| | - Gunnar Brunborg
- Department of Molecular Biology, Norwegian Institute of Public Health, Oslo, Norway
| | - Andrew Collins
- Department of Nutrition, University of Oslo, Oslo, Norway
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31
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Abstract
Human biomonitoring studies aim to identify potential exposures to environmental, occupational, or lifestyle toxicants in human populations and are commonly used by public health decision makers to predict disease risk. The Comet assay measures changes in genomic stability and is one of the most reliable biomarkers to indicate early biological effects and therefore accepted by various governmental regulatory agencies. The appeal of the Comet assay lies in its relative simplicity, rapidity, sensitivity, and economic efficiency. Furthermore, the assay is known for its broad versatility, as it can be applied to virtually any human cell and easily adapted in order to detect particular biomarkers of interest, such as DNA repair capacity or single and double-strand breaks. In a standard experiment, isolated single cells are first embedded in agarose, and then lysed in high-salt solutions in order to remove all cellular contents except the DNA attached to a nuclear scaffold. Subsequent electrophoresis results in accumulation of undamaged DNA sequences at the proximity of the nuclear scaffold, while damaged sequences migrate toward the anode. When visualized with fluorochromes, these migrated DNA fragments resemble a Comet tail and can be quantified for their intensity and shape according to internationally drafted guidelines.
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Affiliation(s)
- Diana Anderson
- Faculty of Life Sciences, University of Bradford, Bradford, UK.
| | - Alok Dhawan
- Nanomaterial Toxicology Group, CSIR-Indian Institute of Toxicology Research, Lucknow, Uttar Pradesh, India
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32
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Klingelfus T, Disner GR, Voigt CL, Alle LF, Cestari MM, Leme DM. Nanomaterials induce DNA-protein crosslink and DNA oxidation: A mechanistic study with RTG-2 fish cell line and Comet assay modifications. CHEMOSPHERE 2019; 215:703-709. [PMID: 30347365 DOI: 10.1016/j.chemosphere.2018.10.118] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/20/2018] [Revised: 10/13/2018] [Accepted: 10/16/2018] [Indexed: 06/08/2023]
Abstract
Genotoxic effects of nanomaterials (NMs) have been controversially reported in literature, and the mode of action (MoA) via DNA oxidation is cited as the main damage caused by them. Evidence of nano-silver as a crosslinker has been previously reported by the present research team in an in vivo fish genotoxicity study. Thus, aiming to confirm the evidence about NMs as crosslinker agent, the present investigation elucidated the genotoxic potential of NMs and their genotoxic MoA through in vitro assay with RTG-2 cells line (rainbow trout gonadal) by exposure to nano-silver (PVP-coated) and nano-titanium. The types and levels of DNA damage were assessed by the Comet assay (standard alkaline, hOGG1-modified alkaline, and two crosslink-modified alkaline versions). It was demonstrated that the use of the standard alkaline Comet assay alone may inaccurately predict the genotoxicity of NMs since oxidative and crosslink DNA damages were also verified in RTG-2 cells when assessed by the modified versions of the alkaline protocol. More importantly, it was confirmed that both nano-silver and nano-titanium acted as DNA-protein crosslinkers through the Comet assay version with proteinase K. As both nano-silver and nano-titanium present a great risk to aquatic life, these findings reinforce the need of genotoxicity testing strategies that encompass the assessment of different types of DNA damage, in order to ensure an accurate prediction of the genotoxic potential of NMs.
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Affiliation(s)
- T Klingelfus
- Genetics Department, Federal University of Paraná, Curitiba, Paraná State, Brazil.
| | - G R Disner
- Genetics Department, Federal University of Paraná, Curitiba, Paraná State, Brazil.
| | - C L Voigt
- Chemistry Department, State University of Ponta Grossa, Ponta Grossa, Paraná State, Brazil.
| | - L F Alle
- Genetics Department, Federal University of Paraná, Curitiba, Paraná State, Brazil.
| | - M M Cestari
- Genetics Department, Federal University of Paraná, Curitiba, Paraná State, Brazil.
| | - D M Leme
- Genetics Department, Federal University of Paraná, Curitiba, Paraná State, Brazil.
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Simon L, Emery B, Carrell DT. Sperm DNA Fragmentation: Consequences for Reproduction. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2019; 1166:87-105. [DOI: 10.1007/978-3-030-21664-1_6] [Citation(s) in RCA: 25] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/04/2022]
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Gobi N, Vaseeharan B, Rekha R, Vijayakumar S, Faggio C. Bioaccumulation, cytotoxicity and oxidative stress of the acute exposure selenium in Oreochromis mossambicus. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 2018; 162:147-159. [PMID: 29990726 DOI: 10.1016/j.ecoenv.2018.06.070] [Citation(s) in RCA: 132] [Impact Index Per Article: 18.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/06/2018] [Revised: 06/19/2018] [Accepted: 06/24/2018] [Indexed: 05/12/2023]
Abstract
Selenium (Se) is an essential trace-element that becomes toxic when present at high concentrations for aquatic organisms. The knowledge about the mechanism of Se toxicity in freshwater ecosystem is still poorly studied. Thus the aim of the present study was to assess the impact of environmentally relevant concentrations of Se toxicity: 5, 10, 25, 50 and 100 µg/L or water only (control) for periods of 96 hour (h) to test for Se accumulation (gill, liver and brain), its effects on enzymatic and non-enzymatic antioxidant defenses (gill and liver), oxidative stress effects on lipid, protein (gill and liver), DNA (liver) and inhibition of AchE (brain) activity were measured in Mozambique tilapia, Oreochromis mossambicus. Our result showed that Se accumulation was observed in the gill, liver and brain tissues of fish exposed to different concentrations and accumulation varied upon different tissues. Enzymatic (SOD, CAT, GPx and GST) and non-enzymatic (GSH and MT) antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-s-transferase (GST) were significantly increased after 96 h exposure of higher concentrations Se in the gill and liver tissue with the exception of GST activity was significantly inhibited in liver after 96 h exposure of higher concentrations of Se. In contrast, catalase (CAT) activities were inhibited for both tissues of Se exposure at 96 h. Reduced glutathione (GSH) and Metallothionein (MT) levels were increased in the gill and liver tissues after exposure to Se for 96 h. We also observed that Se affected antioxidant defense, increasing oxidative stress indicator of lipid peroxidation (LPO) and protein carbonyl (PCO) in gill and liver tissues of fish exposed to Se for 96 h at the concentration dependent manner. Increased DNA damage scores observed in liver tissue of fish exposed to Se for concentrations dependent manner, indicating potential of Se on fish. We also observed inhibition of acetylcholine esterase (AchE) activity in brain tissue of fish exposed to Se for higher concentrations. The changes in these parameters can be used as suitable biomarkers for monitoring the toxicity of Se in the aquatic environment.
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Affiliation(s)
- Narayanan Gobi
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Alagappa University, Science Campus 6(th) Floor, Karaikudi 630004, Tamil Nadu, India
| | - Baskaralingam Vaseeharan
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Alagappa University, Science Campus 6(th) Floor, Karaikudi 630004, Tamil Nadu, India.
| | - Ravichandran Rekha
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Alagappa University, Science Campus 6(th) Floor, Karaikudi 630004, Tamil Nadu, India
| | - Sekar Vijayakumar
- Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Alagappa University, Science Campus 6(th) Floor, Karaikudi 630004, Tamil Nadu, India
| | - Caterina Faggio
- Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina-Viale Ferdinando Stagno d'Alcontres, 31 98166 S.Agata-Messina, Italy
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Kim GY. What should be done for men with sperm DNA fragmentation? Clin Exp Reprod Med 2018; 45:101-109. [PMID: 30202739 PMCID: PMC6125150 DOI: 10.5653/cerm.2018.45.3.101] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2018] [Accepted: 08/24/2018] [Indexed: 12/25/2022] Open
Abstract
In an age when a small quantity of sperm can lead to pregnancy through in vitro fertilization or intracytoplasmic sperm injection, selecting healthy sperm is important. Sperm DNA fragmentation (SDF) is known to be higher in infertile men. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and the alkaline comet test are SDF tests that directly measure DNA damage and have shown closer correlations with assisted reproduction results than indirect tools such as the sperm chromatin structure assay or the sperm chromatic dispersion test. It is difficult; however, to endorse a single test as the best test overall; instead, it is best to select a testing method based on each patient's clinical condition and goals. In a couple struggling with infertility, if the male partner has a high level of SDF, he should aim to decrease SDF through lifestyle modifications, antioxidant treatment, and ensuring an appropriate duration of abstinence, and physicians need to treat the underlying diseases of such patients. If sperm DNA damage continues despite the patient's and physician's efforts, other methods, such as micromanipulation-based sperm selection or testicular sperm extraction, should be used to select healthy sperm with nuclear DNA integrity.
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Affiliation(s)
- Gi Young Kim
- Department of Urology, Mizmedi Hospital, Seoul, Korea
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Rodriguez A, Duez P, Dedeken L, Cotton F, Ferster A. Hydroxyurea (hydroxycarbamide) genotoxicity in pediatric patients with sickle cell disease. Pediatr Blood Cancer 2018. [PMID: 29512872 DOI: 10.1002/pbc.27022] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
BACKGROUND Hydroxyurea (HU) reduces the severity of sickle cell disease (SCD) in children; nevertheless, its long-term safety is an important concern. This paper evaluates HU genotoxicity at dose ≤ 30 mg/kg/day after over 2 years of treatment. PROCEDURE The study included 76 children: 32 SCD patients treated with HU, 27 SCD patients not treated with HU, and 17 unaffected children. HU patients were classified as good or poor responders according to their clinical response. Comet assay allows the comparison of DNA damage between both groups of patients and unaffected children. Maximal concentration (Cmax ) of HU in plasma was determined after drug administration. RESULTS Mean values of DNA in the comet tail were 5.13 ± 6.84 for unaffected children, 5.80 ± 7.78 for patients with SCD treated with HU, and 5.61 ± 6.91 for patients with SCD not treated with HU. Significant differences were observed between unaffected children and children with SCD. No difference was evident between comets from SCD patients treated and not treated with HU. In the case of HU, mean DNA in the comet tail was significantly lower in good responders than in poor responders: 5.54 ± 7.77 and 6.69 ± 8.43, respectively. Mean Cmax value on plasma was 39.08 ± 15.65 mg/l; N = 31. CONCLUSIONS SCD increases, slightly but significantly, DNA damage in lymphocytes from patients with SCD. Patients with SCD treated with HU do not present more nucleoid damage than patients with SCD not treated with HU. Good responders to the HU treatment have significantly less nucleoid damage than poor responders. HU treatment at ≤30 mg/kg/day does not expose patients to a genotoxic plasma concentration.
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Affiliation(s)
- Anar Rodriguez
- Laboratory of Biological and Medical Chemistry, Faculté de Phamacie, Université Libre de Bruxelles (ULB), Brussels, Belgium.,Service of Clinical Chemistry, Laboratoire Hospitalier Universitaire de Bruxelles (LHUB-ULB), Brussels, Belgium
| | - Pierre Duez
- Unit of Therapeutic Chemistry and Pharmacognosy, Faculté de Medicine et Pharmacie, Université de Mons (UMONS), Brussels, Belgium
| | - Laurence Dedeken
- Department of Hematology/Oncology, Hôpital Universitaire des Enfants "Reine Fabiola", Brussels, Belgium
| | - Frédéric Cotton
- Laboratory of Biological and Medical Chemistry, Faculté de Phamacie, Université Libre de Bruxelles (ULB), Brussels, Belgium.,Service of Clinical Chemistry, Laboratoire Hospitalier Universitaire de Bruxelles (LHUB-ULB), Brussels, Belgium
| | - Alina Ferster
- Department of Hematology/Oncology, Hôpital Universitaire des Enfants "Reine Fabiola", Brussels, Belgium
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Mokra K, Woźniak K, Bukowska B, Sicińska P, Michałowicz J. Low-concentration exposure to BPA, BPF and BPAF induces oxidative DNA bases lesions in human peripheral blood mononuclear cells. CHEMOSPHERE 2018; 201:119-126. [PMID: 29518729 DOI: 10.1016/j.chemosphere.2018.02.166] [Citation(s) in RCA: 69] [Impact Index Per Article: 9.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/05/2018] [Revised: 02/16/2018] [Accepted: 02/26/2018] [Indexed: 05/25/2023]
Abstract
Because bisphenol A (BPA) and some of its analogs have been supposed to influence development of cancer, we have assessed the effect of BPA, bisphenol S (BPS), bisphenol F (BPF) and bisphenol AF (BPAF) on DNA bases oxidation, which is a key process in cancer initiation. The analysis was conducted on human peripheral blood mononuclear cells (PBMCs), which are very useful model to assess genotoxic potential of various toxicants in different cell types. In order to determine oxidative damage to DNA pyrimidines and purines, alkaline version of the comet assay with DNA glycosylases, i.e. endonuclease III (Nth) and human 8-oxoguanine DNA glycosylase (hOGG1) was used. PBMCs were exposed to BPA or its analogs in the concentrations of 0.01, 0.1 and 1 μg/mL for 4 h and 0.001, 0.01 and 0.1 μg/mL for 48 h. We have observed that BPA, BPS, BPF and particularly BPAF caused oxidative damage to DNA pyrimidines and more strongly to purines in human PBMCs. The results have also shown that BPS, which is the most commonly used as a substitute for BPA in the manufacture induced definitely the smallest oxidative DNA bases lesions in PBMCs. Moreover, we have noticed that BPA, BPF and BPAF caused DNA damage at very low concentration of 1 ng/mL.
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Affiliation(s)
- Katarzyna Mokra
- Department of Biophysics of Environmental Pollution, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143 St., 90-001 Łódź, Poland
| | - Katarzyna Woźniak
- Department of Molecular Genetics, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143 St., 90-001 Łódź, Poland
| | - Bożena Bukowska
- Department of Biophysics of Environmental Pollution, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143 St., 90-001 Łódź, Poland
| | - Paulina Sicińska
- Department of Biophysics of Environmental Pollution, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143 St., 90-001 Łódź, Poland
| | - Jaromir Michałowicz
- Department of Biophysics of Environmental Pollution, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143 St., 90-001 Łódź, Poland.
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Parhiz H, Khoshnejad M, Myerson JW, Hood E, Patel PN, Brenner JS, Muzykantov VR. Unintended effects of drug carriers: Big issues of small particles. Adv Drug Deliv Rev 2018; 130:90-112. [PMID: 30149885 PMCID: PMC6588191 DOI: 10.1016/j.addr.2018.06.023] [Citation(s) in RCA: 49] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2018] [Revised: 06/11/2018] [Accepted: 06/26/2018] [Indexed: 02/06/2023]
Abstract
Humoral and cellular host defense mechanisms including diverse phagocytes, leukocytes, and immune cells have evolved over millions of years to protect the body from microbes and other external and internal threats. These policing forces recognize engineered sub-micron drug delivery systems (DDS) as such a threat, and react accordingly. This leads to impediment of the therapeutic action, extensively studied and discussed in the literature. Here, we focus on side effects of DDS interactions with host defenses. We argue that for nanomedicine to reach its clinical potential, the field must redouble its efforts in understanding the interaction between drug delivery systems and the host defenses, so that we can engineer safer interventions with the greatest potential for clinical success.
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Affiliation(s)
- Hamideh Parhiz
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
| | - Makan Khoshnejad
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
| | - Jacob W Myerson
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
| | - Elizabeth Hood
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
| | - Priyal N Patel
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
| | - Jacob S Brenner
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
| | - Vladimir R Muzykantov
- Department of Pharmacology, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Center for Targeted Therapeutics and Translational Nanomedicine (CT3N), University of Pennsylvania, Philadelphia, PA, USA.
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Physical principles and new applications of comet assay. Biophys Chem 2018; 238:1-7. [PMID: 29704770 DOI: 10.1016/j.bpc.2018.04.003] [Citation(s) in RCA: 30] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2018] [Revised: 04/14/2018] [Accepted: 04/15/2018] [Indexed: 11/21/2022]
Abstract
The comet assay is a sensitive method to assess DNA damages in single cells. The approach consists of an analysis of electrophoretic migration of DNA from nucleoids obtained after cell lysis in a thin layer of agarose. Although the method is widely used the physical mechanisms of DNA track formation remained to be rather elusive for a long time. This review is devoted to our recent results pertaining to this subject, using an original approach based on the kinetic measurements of the comet formation. We argue that linear DNA fragments give an essential contribution into the tail formation in the alkaline conditions and, at neutral pH, when the level of DNA damages is very high. On the other hand, in the neutral comet assay at low levels of DNA damages (and also in the case of undamaged cells) the tail is formed by extended DNA loops. These loops are about the same as chromatin loops in the cell nuclei. Kinetic measurements in the comet assay give an opportunity to investigate the topology of the loops and large-scale features of the loop domain organization (and re-organization) in nucleoids obtained from different cell types.
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40
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Activation of PXR inhibits LPS-induced NF-κB activation by increasing IκBα expression in HepG2 cells. Mol Cell Toxicol 2018. [DOI: 10.1007/s13273-018-0012-6] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
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Ajima MNO, Pandey PK, Kumar K, Poojary N. Alteration in DNA structure, molecular responses and Na + -K + -ATPase activities in the gill of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) in response to sub-lethal verapamil. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY 2018; 147:809-816. [PMID: 28954371 DOI: 10.1016/j.ecoenv.2017.09.050] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/17/2017] [Revised: 09/16/2017] [Accepted: 09/17/2017] [Indexed: 06/07/2023]
Abstract
The ecotoxicological consequences of residues from pharmaceutical drugs on aquatic biota have necessitated the development of sensitive and reliable techniques to assess the impact of these xenobiotics on aquatic organisms. This study investigated the alteration in DNA structure, molecular responses and the activities of Na+ -K+ -ATPase and antioxidant enzymes in the gill of Nile tilapia, Oreochromis niloticus, exposed to long-term effects at the concentrations (0.14, 0.28 and 0.57mgL-1) of verapamil in static renewal system for 15, 30, 45 and 60 days. Evaluation of DNA structure, using single cell gel electrophoresis, revealed certain degree of DNA damages in the gill in a time and concentration-dependent relationship. Transcription of mRNA of superoxide dismutase (sod), catalase (cat) and heat shock protein (hsp70) genes in the gill of the fish showed the genes were up-regulated. Na+-K+-ATPase activity was inhibited in a concentration and time dependent manner. The indices of oxidative stress biomarkers (lipid peroxidation and carbonyl protein) as well as superoxide dismutase, glutathione peroxidase, glutathione-S-transferase were elevated in the treated fish in comparison to the control. Further, the level of reduced glutathione and catalase activity were inhibited at 0.28mgL-1 after day 30. Long-term exposure to sub-lethal concentration of verapamil can cause DNA damages, molecular effects and oxidative stress in O. niloticus. The biomarkers analysed can be used as early warning signals in environmental biomonitoring and assessment of drug contamination in aquatic ecosystem.
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Affiliation(s)
- Malachy N O Ajima
- Department of Fisheries and Aquaculture Technology, Federal University of Technology, Owerri, Nigeria.
| | - Pramod K Pandey
- College of Fisheries, Central Agriculture University, Agartala, Tripura, India
| | - Kundan Kumar
- Aquatic Environment and Health Management Division, ICAR, Central Institute of Fisheries Education, Mumbai, India
| | - Nalini Poojary
- Aquatic Environment and Health Management Division, ICAR, Central Institute of Fisheries Education, Mumbai, India
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Wang H, Zhang Y, Tian Z, Ma J, Kang M, Ding C, Ming D. Preparation of β-CD-Ellagic Acid Microspheres and Their Effects on HepG2 Cell Proliferation. Molecules 2017; 22:molecules22122175. [PMID: 29292740 PMCID: PMC6149914 DOI: 10.3390/molecules22122175] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2017] [Revised: 11/29/2017] [Accepted: 12/06/2017] [Indexed: 01/17/2023] Open
Abstract
OBJECTIVE In this study, β-cyclodextrin (β-CD) was chosen as the coating for ellagic acid to prepare ellagic acid microspheres, and the effect of microspheres on the growth of HepG2 cells was observed. METHODS Scanning electron microscopy, infrared spectroscopy, and release rate analysis were used to identify the formation of ellagic acid microspheres. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the effect of different concentrations of ellagic acid microspheres on tumor cell proliferation at 6, 12, 24 and 36 h, and cell morphology and quantity were observed using hematoxylin-eosin (HE) staining. Single-cell gel electrophoresis was used to observe the effect of ellagic acid microspheres on the DNA damage of HepG2 cells, and the Olive tail moment and the mRNA expression of tumor suppressor protein gene p53 was measured. RESULTS β-CD could be used as wrapping material of ellagic acid to prepare ellagic acid microspheres. HepG2 cell proliferation could be inhibited by 0.1, 0.3 and 0.5 g/L of ellagic acid microspheres in a dose- and time-dependent manner, and the mechanism of proliferation inhibition was related to DNA damage and cell apoptosis. CONCLUSION Preparing ellagic acid microspheres with β-CD is feasible, and ellagic acid microspheres have potential therapeutic value (anticancer).
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Affiliation(s)
- Hongkai Wang
- College of Life Science, Zaozhuang University, Zaozhuang 277160, China.
| | - Yingxia Zhang
- College of Life Science, Zaozhuang University, Zaozhuang 277160, China.
| | - Zhongjing Tian
- College of Life Science, Zaozhuang University, Zaozhuang 277160, China.
| | - Jing Ma
- College of Medical Science, Zaozhuang Vocational College, Zaozhuang 277800, China.
| | - Meiling Kang
- College of Life Science, Zaozhuang University, Zaozhuang 277160, China.
| | - Chengshi Ding
- College of Life Science, Zaozhuang University, Zaozhuang 277160, China.
| | - Dongfeng Ming
- College of Life Science, Zaozhuang University, Zaozhuang 277160, China.
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Cheng N, Wang Y, Cao W. The Protective Effect of Whole Honey and Phenolic Extract on Oxidative DNA Damage in Mice Lymphocytes Using Comet Assay. PLANT FOODS FOR HUMAN NUTRITION (DORDRECHT, NETHERLANDS) 2017; 72:388-395. [PMID: 28929426 DOI: 10.1007/s11130-017-0634-1] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/07/2023]
Abstract
In this study, the antioxidant activity and the protective effect against hydrogen peroxide-induced DNA damage were assessed for five honeys of different botanical origin. Seven phenolic acids were detected in the honey samples. Ferulic acid was the most abundant phenolic acid detected in longan honey, jujube honey and buckwheat honey. Ellagic acid, p-hydroxybenzoic acid and protocatechuic acid were the main phenolic acids detected in vitex honey. Of all honey samples tested, the highest total phenolic content and antioxidant activity were found in buckwheat honey, whereas the lowest total phenolic content and antioxidant activity were found in locust honey. Treatment with hydrogen peroxide induced a 62% increase in tail DNA in mice lymphocytes, and all studied honeys significantly inhibited this effect (P < 0.05). The buckwheat honey with higher antioxidant capability also exhibited super protective effect than others. Phenolic extracts of honey displayed greater protective effects than whole honey in comet assay. The hydrogen peroxide-generated increase in 8-hydroxy-2-deoxyguanosine (8-OHdG) was effectively inhibited by the honeys studied (P < 0.05). Moreover, a dose-effect relationship between honey concentration and its protective effect was clearly observed in this study. It can be deduced that phenolic acids of honey can penetrate into lymphocytes and protect DNA from oxidative damage by scavenging hydrogen peroxide and/or chelating ferrous ions.
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Affiliation(s)
- Ni Cheng
- Department of Food Science and Engineering, School of Chemical Engineering, Northwest University, 229 North TaiBai Road, Xi'an City, 710069, China
| | - Yuan Wang
- Department of Food Science and Engineering, School of Chemical Engineering, Northwest University, 229 North TaiBai Road, Xi'an City, 710069, China
| | - Wei Cao
- Department of Food Science and Engineering, School of Chemical Engineering, Northwest University, 229 North TaiBai Road, Xi'an City, 710069, China.
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Effect of bovine sperm chromatin integrity evaluated using three different methods on in vitro fertility. Theriogenology 2017; 107:142-148. [PMID: 29154161 DOI: 10.1016/j.theriogenology.2017.11.006] [Citation(s) in RCA: 23] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2017] [Revised: 11/03/2017] [Accepted: 11/03/2017] [Indexed: 01/17/2023]
Abstract
In vitro fertility potential of individual bulls is still relatively uncharacterized. Classical sperm analysis does not include the evaluation of all sperm characteristics and thus, some cell compartments could be neglected. In humans, sperm DNA integrity has already proven to have major influence in embryo development and assisted reproduction techniques successfully. In bovine, some studies already correlated chromatin integrity with field fertility. However, none of those have attempted to relate DNA assessment approaches such as chromatin deficiency (CMA3), chromatin stability (SCSA; AO+) and DNA fragmentation (COMET assay) to predict in vitro bull fertility. To this purpose, we selected bulls with high and low in vitro fertility (n = 6/group), based on embryo development rate (blastocyst/cleavage rate). We then performed CMA3, SCSA test and COMET assay to verify if the difference of in vitro fertility may be related to DNA alterations evaluated by these assays. For the three tests performed, our results showed only differences in the percentage of cells with chromatin deficiency (CMA3+; high: 0.19 ± 0.03 vs low: 0.04 ± 0.04; p = 0.03). No difference for chromatin stability and any of COMET assay categories (grade I to grade IV) was observed between high and low in vitro fertility bulls. A positive correlation between AO + cells and grade IV cells was found. Despite the difference between groups in CMA3 analysis, our results suggest that protamine deficiency in bovine spermatozoa may not have a strong biological impact to explain the difference of in vitro fertility between the bulls used in this study.
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Bielecka-Kowalska A, Czarny P, Wigner P, Synowiec E, Kowalski B, Szwed M, Krupa R, Toma M, Drzewiecka M, Majsterek I, Szemraj J, Sliwinski T, Kowalski M. Ethylene glycol dimethacrylate and diethylene glycol dimethacrylate exhibits cytotoxic and genotoxic effect on human gingival fibroblasts via induction of reactive oxygen species. Toxicol In Vitro 2017; 47:8-17. [PMID: 29107684 DOI: 10.1016/j.tiv.2017.10.028] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/11/2017] [Revised: 10/18/2017] [Accepted: 10/27/2017] [Indexed: 12/25/2022]
Abstract
Although methacrylic acid derivatives in their polymeric form are considered to be safe, insufficient polymerization and the release of monomers due to either mechanical or enzymatical factors can lead to their reaching millimolar concentrations in local tissue. The present study evaluates the effect of two methacrylate monomers - ethylene glycol dimethacrylate (EGDMA) and diethylene glycol dimethacrylate (DEGDMA) - on human gingival fibroblasts (HGFs). Both monomers were found to reduce cells viability in MTT assay, increase apoptosis and cause cell cycle arrest in G1/G0 phase. They also increased intracellular reactive oxygen species (ROS) production as measured by DCFH-DA and DHE probes and increased expression of GPx4 and SOD2. Both monomers increased DNA damage in comet assay. Moreover, HGFs were not able to repair those lesions within 120min of repair incubation. However, the monomers were not found to have any effect on the integrity of isolated plasmids. We postulate that EGDMA and DEGDMA exhibit their cytotoxic and genotoxic properties via increased production of ROS, which cause DNA damage, affect apoptosis, viability and cell cycle. Further studies are needed to better understand the properties of methacrylic acid monomers and to evaluate the risk that they cause for patients, dentists and dental technicians.
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Affiliation(s)
| | - Piotr Czarny
- Department of Medical Biochemistry, Medical University of Lodz, Lodz, Poland
| | - Paulina Wigner
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Ewelina Synowiec
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Bartosz Kowalski
- Department of Maxillofacial Surgery, Medical University of Lodz, Lodz, Poland
| | - Marzena Szwed
- Department of Medical Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Renata Krupa
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Monika Toma
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Malgorzata Drzewiecka
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland
| | - Ireneusz Majsterek
- Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, Lodz, Poland
| | - Janusz Szemraj
- Department of Medical Biochemistry, Medical University of Lodz, Lodz, Poland
| | - Tomasz Sliwinski
- Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
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Simon L, Emery BR, Carrell DT. Review: Diagnosis and impact of sperm DNA alterations in assisted reproduction. Best Pract Res Clin Obstet Gynaecol 2017; 44:38-56. [DOI: 10.1016/j.bpobgyn.2017.07.003] [Citation(s) in RCA: 82] [Impact Index Per Article: 10.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2017] [Revised: 07/11/2017] [Accepted: 07/24/2017] [Indexed: 01/31/2023]
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Frenzilli G, Ryskalin L, Ferrucci M, Cantafora E, Chelazzi S, Giorgi FS, Lenzi P, Scarcelli V, Frati A, Biagioni F, Gambardella S, Falleni A, Fornai F. Loud Noise Exposure Produces DNA, Neurotransmitter and Morphological Damage within Specific Brain Areas. Front Neuroanat 2017; 11:49. [PMID: 28694773 PMCID: PMC5483448 DOI: 10.3389/fnana.2017.00049] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/23/2017] [Accepted: 06/06/2017] [Indexed: 12/11/2022] Open
Abstract
Exposure to loud noise is a major environmental threat to public health. Loud noise exposure, apart from affecting the inner ear, is deleterious for cardiovascular, endocrine and nervous systems and it is associated with neuropsychiatric disorders. In this study we investigated DNA, neurotransmitters and immune-histochemical alterations induced by exposure to loud noise in three major brain areas (cerebellum, hippocampus, striatum) of Wistar rats. Rats were exposed to loud noise (100 dBA) for 12 h. The effects of noise on DNA integrity in all three brain areas were evaluated by using Comet assay. In parallel studies, brain monoamine levels and morphology of nigrostriatal pathways, hippocampus and cerebellum were analyzed at different time intervals (24 h and 7 days) after noise exposure. Loud noise produced a sudden increase in DNA damage in all the brain areas under investigation. Monoamine levels detected at 7 days following exposure were differently affected depending on the specific brain area. Namely, striatal but not hippocampal dopamine (DA) significantly decreased, whereas hippocampal and cerebellar noradrenaline (NA) was significantly reduced. This is in line with pathological findings within striatum and hippocampus consisting of a decrease in striatal tyrosine hydroxylase (TH) combined with increased Bax and glial fibrillary acidic protein (GFAP). Loud noise exposure lasting 12 h causes immediate DNA, and long-lasting neurotransmitter and immune-histochemical alterations within specific brain areas of the rat. These alterations may suggest an anatomical and functional link to explain the neurobiology of diseases which prevail in human subjects exposed to environmental noise.
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Affiliation(s)
- Giada Frenzilli
- Department of Clinical and Experimental Medicine, University of PisaPisa, Italy
| | - Larisa Ryskalin
- Department of Translational Research and New Technologies in Medicine and Surgery, University of PisaPisa, Italy
| | - Michela Ferrucci
- Department of Translational Research and New Technologies in Medicine and Surgery, University of PisaPisa, Italy
| | - Emanuela Cantafora
- Department of Clinical and Experimental Medicine, University of PisaPisa, Italy
| | - Silvia Chelazzi
- Department of Clinical and Experimental Medicine, University of PisaPisa, Italy
| | - Filippo S Giorgi
- Department of Clinical and Experimental Medicine, University of PisaPisa, Italy
| | - Paola Lenzi
- Department of Translational Research and New Technologies in Medicine and Surgery, University of PisaPisa, Italy
| | - Vittoria Scarcelli
- Department of Clinical and Experimental Medicine, University of PisaPisa, Italy
| | - Alessandro Frati
- Istituto di Ricovero e Cura a Carattere Scientifico IRCCS NeuromedIsernia, Italy
| | - Francesca Biagioni
- Istituto di Ricovero e Cura a Carattere Scientifico IRCCS NeuromedIsernia, Italy
| | - Stefano Gambardella
- Istituto di Ricovero e Cura a Carattere Scientifico IRCCS NeuromedIsernia, Italy
| | - Alessandra Falleni
- Department of Clinical and Experimental Medicine, University of PisaPisa, Italy
| | - Francesco Fornai
- Department of Translational Research and New Technologies in Medicine and Surgery, University of PisaPisa, Italy.,Istituto di Ricovero e Cura a Carattere Scientifico IRCCS NeuromedIsernia, Italy
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Dynamic In Vivo Profiling of DNA Damage and Repair after Radiotherapy Using Canine Patients as a Model. Int J Mol Sci 2017; 18:ijms18061176. [PMID: 28587165 PMCID: PMC5485999 DOI: 10.3390/ijms18061176] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/05/2017] [Revised: 05/23/2017] [Accepted: 05/27/2017] [Indexed: 01/22/2023] Open
Abstract
Time resolved data of DNA damage and repair after radiotherapy elucidates the relation between damage, repair, and cell survival. While well characterized in vitro, little is known about the time-course of DNA damage response in tumors sampled from individual patients. Kinetics of DNA damage after radiotherapy was assessed in eight dogs using repeated in vivo samples of tumor and co-irradiated normal tissue analyzed with comet assay and phosphorylated H2AX (γH2AX) immunohistochemistry. In vivo results were then compared (in silico) with a dynamic mathematical model for DNA damage formation and repair. Maximum %DNA in tail was observed at 15–60 min after irradiation, with a rapid decrease. Time-courses of γH2AX-foci paralleled these findings with a small time delay and were not influenced by covariates. The evolutionary parameter search based on %DNA in tail revealed a good fit of the DNA repair model to in vivo data for pooled sarcoma time-courses, but fits for individual sarcoma time-courses suffer from the heterogeneous nature of the in vivo data. It was possible to follow dynamics of comet tail intensity and γH2AX-foci during a course of radiation using a minimally invasive approach. DNA repair can be quantitatively investigated as time-courses of individual patients by integrating this resulting data into a dynamic mathematical model.
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Pandey PK, Ajima MNO, Kumar K, Poojary N, Kumar S. Evaluation of DNA damage and physiological responses in Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) exposed to sub-lethal diclofenac (DCF). AQUATIC TOXICOLOGY (AMSTERDAM, NETHERLANDS) 2017; 186:205-214. [PMID: 28324828 DOI: 10.1016/j.aquatox.2017.03.007] [Citation(s) in RCA: 32] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/23/2016] [Revised: 03/03/2017] [Accepted: 03/04/2017] [Indexed: 06/06/2023]
Abstract
The frequent bioaccumulation of pharmaceuticals in the aquatic ecosystem has raised a concern about their possible ecotoxicological consequences. DNA damage, haematological changes and activities of oxidative stress enzymes in Nile tilapia, Oreochromis niloticus in response to diclofenac (DCF) exposure were investigated for up to 60 days at the concentrations of 0.17, 0.34 and 0.68mgL-1 in the fish liver. Evaluation of genotoxic effects of the drug in the liver, using single-cell gel electrophoresis, showed DNA damage on exposure at the concentrations of 0.34 and 0.68mgL-1 after day 30. Compared with the control, there was a reduction in haemoglobin and red blood cell counts with a significant increase (p<0.05) in white blood cell counts, mean corpuscular volume and mean corpuscular haemoglobin level after day 30 at 0.34 and 0.68mgL-1. The levels of pack cell volume, red cell distribution width and mean corpuscular haemoglobin concentration were not significant (p>0.05) between the exposed group and the control. The indices of hepatic oxidative stress biomarkers, including lipid peroxidation and carbonyl protein, showed elevated level, depicting a positive correlation with both time and concentration. More so, activity of catalase was inhibited while reduced glutathione level decreased in the liver tissue. There was increase in the activities of superoxide dismutase, glutathione peroxidase and glutathione-S-transferase after 30 days at 0.34mgL-1. Further, activity of Na+-K+-ATPase in the tissue was significantly inhibited (p<0.05) at the end of 60 days. Prolonged exposure to diclofenac at sub-lethal concentration can cause both DNA and oxidative damages in O. niloticus, suggesting the use of oxidative stress biomarkers as early warning signals in environmental monitoring of residual pharmaceutical and assessment.
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Affiliation(s)
- Pramod K Pandey
- College of Fisheries, Central Agriculture University, Agartala, Tripura, India
| | - Malachy N O Ajima
- Department of Fisheries and Aquaculture Technology, Federal University of Technology, Owerri, Nigeria; Aquatic Environment and Health Management Division, ICAR-Central Institute of Fisheries Education, Mumbai, India.
| | - Kundan Kumar
- Aquatic Environment and Health Management Division, ICAR-Central Institute of Fisheries Education, Mumbai, India
| | - Nalini Poojary
- Aquatic Environment and Health Management Division, ICAR-Central Institute of Fisheries Education, Mumbai, India
| | - Saurav Kumar
- Aquatic Environment and Health Management Division, ICAR-Central Institute of Fisheries Education, Mumbai, India
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Szejk M, Poplawski T, Czubatka-Bienkowska A, Olejnik AK, Pawlaczyk-Graja I, Gancarz R, Zbikowska HM. A comparative study on the radioprotective potential of the polyphenolic glycoconjugates from medicinal plants of Rosaceae and Asteraceae families versus their aglycones. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY 2017; 171:50-57. [PMID: 28475935 DOI: 10.1016/j.jphotobiol.2017.04.027] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/25/2016] [Revised: 04/11/2017] [Accepted: 04/24/2017] [Indexed: 01/30/2023]
Abstract
Radioprotective potential of the polyphenolic glycoconjugates, isolated from flowers of Sanguisorba officinalis L. (So) and Erigeron canadensis L. (Ec), and from leaves of Fragaria vesca L. (Fv) and Rubus plicatus Whe. Et N. E. (Rp) as well as their aglycones (SoA, EcA, FvA and RpA, respectively), against γ-radiation-induced lipid peroxidation in human plasma and DNA damage in lymphocytes, were investigated in vitro. These properties were assessed by measuring the concentration of thiobarbituric acid reactive substances (TBARS) and using the alkaline comet assay, and were compared to the protective effects of rutin (R) and quercetin (Q). Cytotoxicity of the glycoconjugates/aglycones towards L929 mouse fibroblasts and human lymphocytes were also measured. Plant products from S. officinalis, similar to Q, were able to reduce the most radiation-induced lipid peroxidation as well as DNA damage and extent of oxidative damage to the DNA basis. Contrary to the pure flavonoids, where Q was shown to be significantly more effective than its glycoside R, the results did not show more benefit with application of SoA/EcA over So/Ec in terms of lipid peroxidation inhibition. Moreover, glycoconjugates Ec and So showed much higher capacity in protecting lymphocytes against radiation-induced genotoxicity which may suggest that between the polyphenolic and polysaccharide parts exist some synergistic effects. There were no significant differences between Fv versus FvA or Rp versus RpA in terms of the provided radioprotection. Summarizing, plant glycoconjugates isolated by the multi-step method offered sufficient radioprotection. In addition, they possess many advantages, compared to the synthetic polyphenolic compounds or the plant extracts, such as water-solubility and minor toxicity.
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Affiliation(s)
- Magdalena Szejk
- Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
| | - Tomasz Poplawski
- Department of Molecular Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
| | - Anna Czubatka-Bienkowska
- Department of Molecular Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland
| | - Alicja Klaudia Olejnik
- Chemistry Department, Institute of Applied Radiation Chemistry, Lodz University of Technology, Wroblewskiego 15, 93-590 Lodz, Poland
| | - Izabela Pawlaczyk-Graja
- Division of Organic and Pharmaceutical Technology, Faculty of Chemistry, Wroclaw University of Science and Technology, Wybrzeze Wyspianskiego 29, 50-370 Wroclaw, Poland
| | - Roman Gancarz
- Division of Organic and Pharmaceutical Technology, Faculty of Chemistry, Wroclaw University of Science and Technology, Wybrzeze Wyspianskiego 29, 50-370 Wroclaw, Poland
| | - Halina Malgorzata Zbikowska
- Department of General Biochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland.
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