Single cell gel electrophoresis, or the comet assay, was devised as a sensitive method for detecting DNA strand breaks, at the level of individual cells. A simple modification, incorporating a ...digestion of DNA with a lesion-specific endonuclease, makes it possible to measure oxidised bases.
With the inclusion of formamidopyrimidine DNA glycosylase to recognise oxidised purines, or Nth (endonuclease III) to detect oxidised pyrimidines, the comet assay has been used extensively in human biomonitoring to monitor oxidative stress, usually in peripheral blood mononuclear cells.
There is evidence to suggest that the enzymic approach is more accurate than chromatographic methods, when applied to low background levels of base oxidation. However, there are potential problems of over-estimation (because the enzymes are not completely specific) or under-estimation (failure to detect lesions that are close together). Attempts have been made to improve the inter-laboratory reproducibility of the comet assay.
In addition to measuring DNA damage, the assay can be used to monitor the cellular or in vitro repair of strand breaks or oxidised bases. It also has applications in assessing the antioxidant status of cells. In its various forms, the comet assay is now an invaluable tool in human biomonitoring and genotoxicity testing. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.
•The comet assay measures DNA breaks at the level of individual cells•Including lesion-specific endonucleases in the comet assay allows detection of DNA oxidation•It is the method of choice when there are low levels of damage as in human cells•As well as DNA damage, antioxidant status and DNA repair can be assessed
A model of murine IgG function is presented in which the co-expression of the IgG subclasses is a central feature, class switching occurs before the commencement of somatic hypermutation, and there ...is little switching between subclasses. It is named the quartet model to emphasize the harmony that comes from the simultaneous presence of the four subclasses. In this model, IgG3 and IgG2b antibodies are particularly important early in the response, when T-cell help may be limiting. IgG3 initiates inflammation through complement fixation, whereas IgG2b provides early FcγR-mediated effector functions. As T-cell help strengthens, IgG2a antibodies increase the power of the response, whereas IgG1 production helps limit the inflammatory drive and limits immunopathology. The model highlights the fact that murine IgG subclasses function quite differently to human IgG subclasses. This allows them to serve the special immunological needs of a species that is vulnerable because of its small size.
The comet assay (single cell gel electrophoresis) is the most common method for measuring DNA damage in eukaryotic cells or disaggregated tissues. The assay depends on the relaxation of supercoiled ...DNA in agarose-embedded nucleoids (the residual bodies remaining after lysis of cells with detergent and high salt), which allows the DNA to be drawn out towards the anode under electrophoresis, forming comet-like images as seen under fluorescence microscopy. The relative amount of DNA in the comet tail indicates DNA break frequency. The assay has been modified to detect various base alterations, by including digestion of nucleoids with a lesion-specific endonuclease. We describe here recent technical developments, theoretical aspects, limitations as well as advantages of the assay, and modifications to measure cellular antioxidant status and different types of DNA repair. We briefly describe the applications of this method in genotoxicity testing, human biomonitoring, and ecogenotoxicology.
Although strategic coalition formation is traditionally modeled using cooperative game theory, behavioral game theorists have repeatedly shown that outcomes predicted by game theory are different ...from those generated by actual human behavior. To further explore these differences, in a cooperative game theory context, we experiment to compare the outcomes resulting from human participants’ behavior to those generated by a cooperative game theory solution mechanism called the core partition. Our experiment uses an interactive simulation of a glove game, a particular type of cooperative game, to collect the participant’s decision choices and their resultant outcomes. Two different glove games are considered, and the outputs from 62 trial games are analyzed. The experiment’s outcomes show that core coalitions, which are coalitions in a core partition, are found in about 42% of games. Though this number may seem low, a trial’s outcome is more complex than whether the human player finds a core coalition or not. Finding the core coalition depends on factors such as the other possible feasible solutions and the payoffs available from these solutions. These factors, and the complexity they generate, are discussed in the paper.
•Unpacking social vulnerability hidden in dynamic context is possible.•The principal component analysis helps identify hidden social vulnerability.•Proposes novel social vulnerability index to floods ...hazards.•Outlines policy implications for disaster risk reduction decision-makers.
Understanding the complexity of vulnerability to disasters, including those triggered by floods, droughts and epidemics is at the heart of disaster risk reduction. Despite its importance in disaster risk reduction, there remains a paucity of approaches that contribute to our understanding of social vulnerability that is hidden in dynamic contextual conditions. The study demonstrates an accessible means to assessing the spatial variation of social vulnerability to flood hazards and related for the context of Muzarabani district in northeast Zimbabwe. The study facilitated local identification with residents of variables contributing to social vulnerability and used the principal component analysis (PCA) technique to develop a social vulnerability index (SoVI). Using ArcMap10.2 geographic information systems (GIS) tool, the study mapped composite SoVI at the ward level. The results showed that Muzarabani district is socially vulnerable to hazards. The social vulnerability is influenced by a variety of economic, social and institutional factors that vary across the wards. Quantifying and visualising social vulnerability in Muzarabani provides useful information for decision makers to support disaster preparedness and mitigation programmes. The approach shows how spatially distributed multivariate vulnerability, as grounded in interpretations at local level, can be quantitatively derived for contexts such as those of Muzarabani. The study findings can inform disaster risk reduction communities and cognate disciplines on quantitative assessments for managing hazard vulnerability where these have hitherto not been developed.
Chronic pain in children is a relatively prevalent cause of functional disability. Contributing factors to this pain are best viewed through the biopsychosocial model. Although evidence is lacking ...for individual aspects of treatment, interdisciplinary care is considered the best treatment approach for children with chronic pain. Interdisciplinary care can include medication management with daily and as-needed medications, physical and occupational therapy focusing on function and movement, and psychological treatment with cognitive-behavioral therapy and acceptance focused treatment. In children with severe pain and disability, intensive interdisciplinary pain treatment may be needed to improve pain and function.
Polyphenols are a very broad group of chemicals, widely distributed in plant foods, and endowed with antioxidant activity by virtue of their numerous phenol groups. They are widely studied as ...putative cancer-protective agents, potentially contributing to the cancer preventive properties of fruits and vegetables. We review recent publications relating to human trials, animal experiments and cell culture, grouping them according to whether polyphenols are investigated in whole foods and drinks, in plant extracts, or as individual compounds. A variety of assays are in use to study genetic damage endpoints. Human trials, of which there are rather few, tend to show decreases in endogenous DNA damage and protection against DNA damage induced ex vivo in blood cells. Most animal experiments have investigated the effects of polyphenols (often at high doses) in combination with known DNA-damaging agents, and generally they show protection. High concentrations can themselves induce DNA damage, as demonstrated in numerous cell culture experiments; low concentrations, on the other hand, tend to decrease DNA damage.
Abstract
The comet assay is widely used in studies on genotoxicity testing, human biomonitoring and clinical studies. The simple version of the assay detects a mixture of DNA strand breaks and ...alkali-labile sites; these lesions are typically described as DNA strand breaks to distinguish them from oxidatively damaged DNA that are measured with the enzyme-modified comet assay. This review assesses the association between high-prevalence diseases in high-income countries and DNA damage measured with the comet assay in humans. The majority of case–control studies have assessed genotoxicity in white blood cells. Patients with coronary artery disease, diabetes, kidney disease, chronic obstructive pulmonary disease and Alzheimer’s disease have on average 2-fold higher levels of DNA strand breaks compared with healthy controls. Patients with coronary artery disease, diabetes, kidney disease and chronic obstructive pulmonary disease also have 2- to 3-fold higher levels of oxidatively damaged DNA in white blood cells than controls, although there is not a clear difference in DNA damage levels between the different diseases. Case–control studies have shown elevated levels of DNA strand breaks in patients with breast cancer, whereas there are only few studies on colorectal and lung cancers. At present, it is not possible to assess if these neoplastic diseases are associated with a different level of DNA damage compared with non-neoplastic diseases.
According to a recent evolutionary life history model of development proposed by Ellis, Figueredo, Brumbach, and Schlomer (2009), growing up in harsh versus unpredictable environments should have ...unique effects on life history strategies in adulthood. Using data from the Minnesota Longitudinal Study of Risk and Adaptation, we tested how harshness and unpredictability experienced in early childhood (age 0-5) versus in later childhood (age 6-16) uniquely predicted sexual and risky behavior at age 23. Findings showed that the strongest predictor of both sexual and risky behavior was an unpredictable environment between ages 0 and 5. Individuals exposed to more unpredictable, rapidly changing environments during the first 5 years of life displayed a faster life history strategy at age 23 by having more sexual partners, engaging in more aggressive and delinquent behaviors, and being more likely to be associated with criminal activities. In contrast, exposure to either harsh environments or experiencing unpredictability in later childhood (age 6-16) was, for the most part, not significantly related to these outcomes at age 23. Viewed together, these findings show that unpredictable rather than merely harsh childhood environments exert unique effects on risky behavior later in life consistent with a faster life history strategy. The findings also suggest that there is a developmentally sensitive period for assessing environmental unpredictability during the first 5 years of life. (Contains 8 tables and 5 footnotes.)
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