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JOURNAL ARTICLE
Shun-Qing Liang, Christopher J Walkey, Alexa E Martinez, Qin Su, Mary E Dickinson, Dan Wang, William R Lagor, Jason D Heaney, Guangping Gao, Wen Xue
Genome editing in the lung has the potential to provide long-term expression of therapeutic protein to treat lung genetic diseases. Yet efficient delivery of CRISPR to the lung remains a challenge. The NIH Somatic Cell Genome Editing (SCGE) Consortium is developing safe and effective methods for genome editing in disease tissues. Methods developed by consortium members are independently validated by the SCGE small animal testing center to establish rigor and reproducibility. We have developed and validated a dual adeno-associated virus (AAV) CRISPR platform that supports effective editing of a lox-stop-lox-Tomato reporter in mouse lung airway...
January 5, 2022: Molecular Therapy
#22
JOURNAL ARTICLE
Kang-Sheng Liu, Xiao-Dong Mao, Feng Pan, Rui Fang An
Recent years have seen a rising incidence of male infertility, mostly caused by the decline of sperm quality. The ratio of infertile males to infertile females has escalated from 3:7 in 2013 to current 5:5, which turns male infertility into the research focus of reproductive medicine. This study aimed to clarify the effect of reproductive tract infection by ureaplasma urealyticum (UU) and chlamydia trachomatis (CT) on the DNA integrity and routine semen parameters of infertile males. A retrospective study was performed...
June 28, 2021: Reproductive Biology and Endocrinology: RB&E
#23
REVIEW
Merlin Bittlinger, Johannes Schwietering, Daniel Strech
Somatic cell genome editing (SCGE) is highly promising for therapeutic innovation. This study demonstrates that the majority of 46 preclinical SCGE studies discussed in reviews as particularly promising for clinical translation do not report on key elements for robust and confirmatory research practices: randomization, blinding, sample size calculation, data handling, pre-registration, multi-centric study design, and independent confirmation. We present the here-examined reporting standards and the new National Institutes of Health (NIH) funding criteria for SCGE research as a viable solution to protect this promising field from backlashes...
October 2021: Drug Discovery Today
#24
JOURNAL ARTICLE
Z He, H Xian, M Tang, Y Chen, Z Lian, D Fang, X Peng, D Hu
Methyl tert-butyl ether (MTBE), a widely used gasoline additive and a ubiquitous environmental pollutant in many countries and regions, can cause various kinds of toxic effects on human health. However, the molecular mechanism underlying its toxic effects remains elusive. The present study aimed to explore the cytotoxicity, DNA damage and oxidative damage effects of MTBE on human bronchial epithelial cells (16HBE) and the possible role of DNA polymerase β (pol-β) in this process. RNA interference (RNAi) was used to obtain pol-β gene knocked-down cells (pol-β- )...
December 2021: Human & Experimental Toxicology
#25
JOURNAL ARTICLE
Ambra Dell'Orso, Elena Kuzminsky, Victoria Bermejo-Bermejo, Raquel Ruiz-Checa, Rocío Alonso-Del Amo, Roberta Meschini
Ulmus glabra is a deciduous tree with a wide distribution in the Eurosiberian region. The southernmost populations, in the Mediterranean area, are fragmented in mountain areas which act as a refugium. These small relict populations can act as sentinel of global change, including climate change and impacts of human activities such as air pollution. Besides, tropospheric ozone (O3 ) is an additional stress factor in the Mediterranean region affecting plant physiology and health. Moreover, oxidative stress caused by O3 could increase DNA damage in plants cells...
August 2021: Ecotoxicology
#26
JOURNAL ARTICLE
Nomali Zanele Dlamini, Anou M Somboro, Daniel G Amoako, Isaiah Arhin, Hezekiel M Khumalo, Rene B Khan
The study investigated the toxicogenic effects, molecular mechanisms and proteomic assessment of aflatoxin B1 (AFB1 ) on human renal cells. Hek293 cells were exposed to AFB1 (0-100 μM) for 24 h. The effect on cell viability was assessed using the methylthiazol tetrazolium (MTT) assay, which also produced the half maximal inhibitory concentration (IC50 ) used in subsequent assays. Free radical production was evaluated by quantifying malondialdehyde (MDA) and nitrate concentration, while DNA fragmentation was determined using the single cell gel electrophoresis (SCGE) assay and DNA gel electrophoresis...
June 5, 2021: Environmental Toxicology
#27
REVIEW
Krishanu Saha, Erik J Sontheimer, P J Brooks, Melinda R Dwinell, Charles A Gersbach, David R Liu, Stephen A Murray, Shengdar Q Tsai, Ross C Wilson, Daniel G Anderson, Aravind Asokan, Jillian F Banfield, Krystof S Bankiewicz, Gang Bao, Jeff W M Bulte, Nenad Bursac, Jarryd M Campbell, Daniel F Carlson, Elliot L Chaikof, Zheng-Yi Chen, R Holland Cheng, Karl J Clark, David T Curiel, James E Dahlman, Benjamin E Deverman, Mary E Dickinson, Jennifer A Doudna, Stephen C Ekker, Marina E Emborg, Guoping Feng, Benjamin S Freedman, David M Gamm, Guangping Gao, Ionita C Ghiran, Peter M Glazer, Shaoqin Gong, Jason D Heaney, Jon D Hennebold, John T Hinson, Anastasia Khvorova, Samira Kiani, William R Lagor, Kit S Lam, Kam W Leong, Jon E Levine, Jennifer A Lewis, Cathleen M Lutz, Danith H Ly, Samantha Maragh, Paul B McCray, Todd C McDevitt, Oleg Mirochnitchenko, Ryuji Morizane, Niren Murthy, Randall S Prather, John A Ronald, Subhojit Roy, Sushmita Roy, Venkata Sabbisetti, W Mark Saltzman, Philip J Santangelo, David J Segal, Mary Shimoyama, Melissa C Skala, Alice F Tarantal, John C Tilton, George A Truskey, Moriel Vandsburger, Jonathan K Watts, Kevin D Wells, Scot A Wolfe, Qiaobing Xu, Wen Xue, Guohua Yi, Jiangbing Zhou
The move from reading to writing the human genome offers new opportunities to improve human health. The United States National Institutes of Health (NIH) Somatic Cell Genome Editing (SCGE) Consortium aims to accelerate the development of safer and more-effective methods to edit the genomes of disease-relevant somatic cells in patients, even in tissues that are difficult to reach. Here we discuss the consortium's plans to develop and benchmark approaches to induce and measure genome modifications, and to define downstream functional consequences of genome editing within human cells...
April 2021: Nature
#28
JOURNAL ARTICLE
Hongyi Xian, Meilin Tang, Ying Chen, Zhenwei Lian, Yanfeng Li, Xiaowu Peng, Dalin Hu
This study was conducted to assess the genotoxic potentials of water in the Helong Reservoir, which was designated as a strategic drinking water source by Guangdong Provincial Government of China in October 2016. According to the characteristics of water environment in it, 4 kinds of indigenous fish samples (Labeo rohita, Cirrhinus molitorella, Red tilapia and Oreochromis niloticus) were collected at 6 sampling sites during the period from July to November 2020. Fish from the clean drinking water source of the upper reaches of the Liuxihe Reservoir in Guangzhou was collected as control...
March 24, 2021: Environmental Toxicology and Chemistry
#29
JOURNAL ARTICLE
Noelia Nikoloff, Ana C Carranza Martin, Mariana C Fabra, Cecilia C Furnus
The aim of this study was to evaluate the genotoxic and cytotoxic effects of amitraz (AMZ) on the primary culture of bovine cumulus cells (CC) and oocyte nuclear maturation. Cytotoxicity was evaluated by assessing mitochondrial activity with the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Genotoxicity was estimated using the alkaline single cell gel electrophoresis (SCGE) assay. Apoptosis was detected with the Annexin V-affinity assay. The in vitro maturation test was performed in bovine oocytes...
June 2021: Environmental Science and Pollution Research International
#30
JOURNAL ARTICLE
Wenjing Dong, Erqun Song, Yang Song
The acute liver injury (ALI) and hepatic fibrosis caused by the co-treatment of lipopolysaccharide (LPS)/D-galactosamine (D-GalN) have been extensively studied. However, whether LPS/D-GalN are genotoxic has been left unknown. In this study, male mice were divided into eight groups with eight animals in each group. For acute challenge of LPS/D-GalN, the mice in each group received a combination of LPS/D-GalN via intraperitoneal injection at the dose of 25 μg/kg/250 mg/kg, 25 μg/kg/500 mg/kg, or 50 μg/kg/500 mg/kg body weight...
January 18, 2021: Scientific Reports
#31
JOURNAL ARTICLE
Evgenii P Khizhnyak, Nikolay P Sirota, Elena A Kuznetsova, Larisa N Khizhnyak, Tatyana V Sirota
Alkaline version of the single-cell gel electrophoresis (Comet assay) is widely used in toxicological, environmental, and monitoring studies to assess the DNA damage levels in individual cells. The change in the temperature of the electrophoretic solution is one of the reasons leading to interlaboratory variation of Comet assay results. In this work, changes of surface temperature of the solution during electrophoresis were studied using technique of real-time thermal imaging. It has been found that the electrophoresis is accompanied by nonuniform temperature rise in different areas of the electrophoretic chamber...
May 2021: Electrophoresis
#32
JOURNAL ARTICLE
Zhisong Cui, Xiao Luan, Shujun Li, Xilong Zhao, Zhongting Lin, Jianjun Li, Wei Gao, Li Zheng, Zhengzhao Ma, Jianghao Xie
An enormous amount of oil-containing drill cuttings have been produced by the marine oil and gas industry. The environmental impacts of discharged drilling waste have been extensively studied. However, there is still an urgent need to develop alternative methods to identify the genotoxicity of untreated and treated drill waste in a timely manner before it is discharged. In this study, we developed a relatively rapid, sensitive, and accurate genotoxicity-detection method using Comet assay and the marine benthic goby Mugilogobius chulae...
January 15, 2021: Ecotoxicology and Environmental Safety
#33
JOURNAL ARTICLE
Erik Bivehed, Björn Hellman
In the present paper, we present a substantially revised protocol of the widely used SCGE assay performed under alkaline conditions. In our updated version of the comet assay, which we call the Flash-comet, LiOH is used instead of NaOH during the unwinding and electrophoresis. This allows a higher voltage during the electrophoresis (5 V/cm instead of 0.7 V/cm), making it possible to reduce the unwinding time from 20 to 40 to 2.5 min, and the electrophoresis time from 10 to 20 to 1 min. Still, the Flash-comet was found to detect DNA strand breaks and alkali-labile sites with a higher degree of sensitivity than the conventional protocol in cells that had been exposed to H2 O2 or ionizing radiation...
2020: MethodsX
#34
JOURNAL ARTICLE
Zhendong Wu, Dedong Wang, Jiguo Wu, Ying Guan, Yunxia Jiang, Yi Zhong, Guoxia Zhang
Many organic pollutants attract public health concern due to their genotoxicity. To investigate the genotoxicity of organic matter in surface water of the Pearl River Delta (PRD). Organic substances of 24 samples (dry and wet season) from North River, West River and East River were extracted from 60 L source water by XAD-2 macroporous resin. DNA damage effect of organic extracts was tested in human derived liver cells (HL-7702), using single cell gel electrophoresis (SCGE) assay. The results showed that 100% organic extracts (24/24) could induce DNA damage in HL-7702 cells when the concentration was above 1...
November 14, 2020: Ecotoxicology
#35
JOURNAL ARTICLE
Ehsan H Abu Zeid, Nabela I El Sharkawy, Gihan G Moustafa, Abeer M Anwer, Ahmed G Al Nady
The present study investigated the alleviating role of camel milk (CM) in the mitigation of fenpropathrin (FNP) type II pyrethroid induced oxidative stress, alterations of hepatic (CYP1A1) mRNA expression pattern, and DNA damage using the alkaline comet assay (SCGE) in male rats. Sixty male Sprague-Dawley rats were separated into six groups (n = 10): 1st control (C), 2nd corn oil (CO), 3rd (CM): gavaged CM 2ml/rat, 4th (FNP): gavaged FNP 7.09 mg/kg body weight (BW), 5th (FNP pro/co-treated): gavaged CM firstly for 15 days, then CM + FNP by the same mentioned doses and route, 6th (FNP + CM co-treated): gavaged FNP firstly followed by CM by the same mentioned doses and route...
September 16, 2020: Ecotoxicology and Environmental Safety
#36
JOURNAL ARTICLE
Shuying Yang, Munir Humayun, Vincent J M Salters
Mid-oceanic ridge basalts (MORBs) are depleted in incompatible elements, but ridge segments far from mantle plumes frequently erupt chemically enriched MORBs (E-MORBs). Two major explanations of E-MORBs are that these basalts are generated by the melting of entrained recycled crust (pyroxenite) beneath ridges or by the melting of refertilized peridotites. These two hypotheses can be discriminated with compatible element abundances from Sc to Ge, here termed the ScGe elements. Here, we demonstrate that E-MORBs have systematically lower Ge/Si and Sc contents and slightly higher Fe/Mn and Nb/Ta ratios than depleted MORBs (D-MORBs) due to the mixing of low-degree pyroxenite melts...
June 2020: Science Advances
#37
REVIEW
Renuka Maurya, Alok Kumar Pandey
Tetrahymena is a single-cell eukaryotic organism present in all aquatic environments and can easily be maintained in laboratory conditions in a cost-effective manner. This review gives a brief description of the physiology of Tetrahymena, culture handling, and maintenance of Tetrahymena species. The review article focuses on various toxicological bioassays at different biological organizational (biochemical, individual, population, and community) levels. Furthermore, some techniques such as single cell gel electrophoresis (SCGE) and microcalorimetry assay are also available to investigate the effect of xenobiotics on the integrity of DNA and metabolic state of Tetrahymena species respectively...
June 8, 2020: Science of the Total Environment
#38
JOURNAL ARTICLE
Ying Qiu, Hua Yang, Chunyuan Li, Changlong Xu
With the increasing incidence of male infertility, routine detection of semen is insufficient to accurately assess male fertility. Infertile men, who have lower odds of conceiving naturally, exhibit high levels of sperm DNA fragmentation (SDF). The mechanisms driving SDF include abnormal spermatogenesis, oxidative stress damage, and abnormal sperm apoptosis. As these factors can induce SDF and subsequent radical changes leading to male infertility, detection of the extent of SDF has become an efficient routine method for semen analysis...
April 22, 2020: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
#39
JOURNAL ARTICLE
Yu Zhang, Jin-Long Lai, Xiao-Hui Ji, Xue-Gang Luo
The aim of this study was to evaluate the ecotoxic effect of high concentration cesium (Cs) exposure on plant root growth and its toxicological mechanism. The radicle of broad bean (Vicia faba) was selected as experimental material. The cytotoxic and genotoxic effects of plants exposed to different Cs levels (0.19-1.5 mM) for 48 h were evaluated using scanning electron microscopy (SEM), X-ray fluorescence (XRF) analysis, single-cell gel electrophoresis (SCGE) and random amplified polymorphic DNA (RAPD) assays...
April 17, 2020: Ecotoxicology and Environmental Safety
#40
JOURNAL ARTICLE
Lonnie P Swift, Lianne Castle, Peter J McHugh
DNA interstrand cross-links (ICLs) are an extremely toxic form of DNA damage that cells experience upon exposure to natural metabolites. Moreover, ICLs are cytotoxic lesions produced by a range of clinically important anticancer agents. Therefore, improving our understanding of ICL induction and processing has important implications in biology and medicine. The sensitive detection of ICLs in mammalian cells is challenging but has been aided by the development of a modified form of the single-cell gel electrophoresis (SCGE) assay, also known as the "comet assay...
2020: Methods in Molecular Biology
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