Biomarkers of oxidative stress: methods and measures of oxidative DNA damage (COMET assay) and telomere shortening

Oxidative stress is fast becoming the nutritional and medical buzzword for the twenty-first century. The theoretical importance of oxidative stress in diabetes is highlighted by its potential double impact on metabolic dysfunction on one hand and the vascular system on the other hand. The new concept of oxidative stress, being an important trigger in the onset and progression of diabetes and its complications, emphasizes the need for measurement of markers of oxidation to assess the degree of oxidative stress. While we have been routinely measuring biomarkers in our molecular epidemiology projects, here we discuss the utility of two assays, (a) DNA damage assessment by COMET measurement and (b) telomere length measurement. As DNA damage is efficiently repaired by cellular enzymes, its measurement gives a snapshot view of the level of oxidative stress. The protocol allows for measurement of oxidative DNA damage (FPG-sensitive DNA strand breaks). Telomere length measured by Southern blotting technique allows one to estimate the chronic burden of oxidative stress at the molecular level and is now considered as biomarker of biological aging

Artifacts associated with the measurement of oxidized DNA bases.

In this paper we review recent aspects of the measurement of oxidized DNA bases, currently a matter of debate. There has long been an interest in the determination of the level of oxidized bases in cellular DNA under both normal and oxidative stress conditions. In this respect, the situation is confusing because variations that may be as large as two orders of magnitude have been reported for the yield of the formation of 8-oxo-7,8-dihydroguanine (8-oxoGua) in similar DNA samples. However, recent findings clearly show that application of several assays like gas chromatography-mass spectrometry (GC-MS) and -32P--postlabeling may lead to a significant overestimation of the level of oxidized bases in cellular DNA. In particular, the silylation step, which is required to make the samples volatile for the GC-MS analysis, has been shown to induce oxidation of normal bases at the level of about one oxidized base per 10(4) normal bases. This has been found to be a general process that applies in particular to 8-oxoGua, 8-oxo-7, 8-dihydroadenine,5-hydroxycytosine, 5-(hydroxymethyl)uracil, and 5-formyluracil. Interestingly, prepurification of the oxidized bases from DNA hydrolysate prior to the derivatization reaction prevents artefactual oxidation. Under these conditions, the level of oxidized bases measured by GC-MS is similar to that obtained by HPLC associated with electrochemical detection (HPLC-EC). It should be added that the level of 8-oxo-7,8-dihydro-2;-deoxyguanosine in control cellular DNA has been found to be about fivefold lower than in earlier HPLC-EC measurements by using appropriate conditions of extraction and enzymatic digestion of DNA. Similar conclusions were reached by measuring formamidopyrimidine-DNA glycosylase sensitive sites as revealed by the single cell gel electrophoresis (comet) assay

The Advertisement Value of Transformational & Informational Appeal on Company Facebook Pages

The advertisement value of Facebook is an under-developed area of social media research. Transformational and informational advertising appeal has yet to be studied as it relates to social media. This paper utilizes established classification and measurement scales from marketing literature to classify companies and their Facebook posts and measures the advertisement value of these posts. The study uses a sample of 100 companies from the 2015 Fortune 500 list. Results indicate that posts with transformational appeal are more engaging to the consumer than informational appeal; however, posts with informational appeal have greater advertisement value for the company. The results also indicate there is no relationship between type of company and type of appeal used by companies

Effectiveness, cost-effectiveness and cost-benefit of a single annual professional intervention for the prevention of childhood dental caries in a remote rural Indigenous community

Background The aim of the study is to reduce the high prevalence of tooth decay in children in a remote, rural Indigenous community in Australia, by application of a single annual dental preventive intervention. The study seeks to (1) assess the effectiveness of an annual oral health preventive intervention in slowing the incidence of dental caries in children in this community, (2) identify the mediating role of known risk factors for dental caries and (3) assess the cost-effectiveness and cost-benefit of the intervention. Methods/design The intervention is novel in that most dental preventive interventions require regular re-application, which is not possible in resource constrained communities. While tooth decay is preventable, self-care and healthy habits are lacking in these communities, placing more emphasis on health services to deliver an effective dental preventive intervention. Importantly, the study will assess cost-benefit and cost-effectiveness for broader implementation across similar communities in Australia and internationally. Discussion There is an urgent need to reduce the burden of dental decay in these communities, by implementing effective, cost-effective, feasible and sustainable dental prevention programs. Expected outcomes of this study include improved oral and general health of children within the community; an understanding of the costs associated with the intervention provided, and its comparison with the costs of allowing new lesions to develop, with associated treatment costs. Findings should be generalisable to similar communities around the world. The research is registered with the Australian New Zealand Clinical Trials Registry (ANZCTR), registration number ACTRN12615000693527; date of registration: 3rd July 2015

Derivation of consistent hard rock (1000<Vs<3000 m/s) GMPEs from surface and down-hole recordings: Analysis of KiK-net data

A key component in seismic hazard assessment is the estimation of ground motion for hard rock sites, either for applications to installations built on this site category, or as an input motion for site response computation. Empirical ground motion prediction equations (GMPEs) are the traditional basis for estimating ground motion while VS30 is the basis to account for site conditions. As current GMPEs are poorly constrained for VS30 larger than 1000 m/s, the presently used approach for estimating hazard on hard rock sites consists of “host-to-target” adjustment techniques based on VS30 and κ0 values. The present study investigates alternative methods on the basis of a KiK-net dataset corresponding to stiff and rocky sites with 500 < VS30 < 1350 m/s. The existence of sensor pairs (one at the surface and one in depth) and the availability of P- and S-wave velocity profiles allow deriving two “virtual” datasets associated to outcropping hard rock sites with VS in the range [1000, 3000] m/s with two independent corrections: 1/down-hole recordings modified from within motion to outcropping motion with a depth correction factor, 2/surface recordings deconvolved from their specific site response derived through 1D simulation. GMPEs with simple functional forms are then developed, including a VS30 site term. They lead to consistent and robust hard-rock motion estimates, which prove to be significantly lower than host-to-target adjustment predictions. The difference can reach a factor up to 3–4 beyond 5 Hz for very hard-rock, but decreases for decreasing frequency until vanishing below 2 Hz

MV-QUAL: A tool for understanding decisions to purchase virtual products in the Metaverse

This paper develops the MV-QUAL (Metaverse Quality) scale and explores the various factors that may impact consumers’ perceptions of quality and value when purchasing virtual products in a Metaverse world. Utilizing Flow Theory and SERVQUAL as theoretical lenses to conduct our research, a framework was developed and tested, demonstrating how various factors are interconnected and impact consumers’ shopping experiences in the Metaverse. Data was collected from a national sample of 322 Metaverse users in the United States and analyzed using partial least squares structural equation modeling in SmartPLS, complemented by a neural network causal analysis in Neusrel. The results demonstrated that several factors impact Metaverse store quality and perceived virtual product value, including control, vividness, avatar design, rarity, and resale value. Subsequently, Metaverse store quality and perceived virtual product value positively impact the intention to purchase virtual products in Metaverse. This research contributes to the advancement of understanding consumer behavior in the Metaverse and adds to the emerging marketing literature on the Metaverse. Furthermore, the MV-QUAL scale developed serves as a practical tool that both marketing researchers and practitioners can use to enhance understanding and discover new ways to improve the Metaverse consumer experience

Palaeoceanographic implications of new and revised bio-chronostratigraphic constraints from the Profitis Ilias Unit (Rhodes, Greece)

Middle Oxfordian-early Kimmeridgian radiolaria, extracted from the top of radiolarites of Profitis Ilias unit (Rhodes island), suggest that the latter are essentially Middle Jurassic in age and the overlying siliceous shales Late Jurassic. The previously identified Calpionellid horizon at the top of Profitis Ilias siliceous marls is now regarded as early Valanginian in age. The above chronostratigraphic constraints allow tentative correlations to be made between Profitis Ilias and Pindos-Olonos sedimentary units. Finally, the palaeoceanographic significance of the studied series in Rhodes and potentially similar pelagic sequences in the Marmaris area of Turkey are discussed

Palaeoceanographic implications of new and revised bio-chronostratigraphic constraints from the Profitis Ilias Unit (Rhodes, Greece)

Middle Oxfordian-early Kimmeridgian radiolaria, extracted from the top of radiolarites of Profitis Ilias unit (Rhodes island), suggest that the latter are essentially Middle Jurassic in age and the overlying siliceous shales Late Jurassic. The previously identified Calpionellid horizon at the top of Profitis Ilias siliceous marls is now regarded as early Valanginian in age. The above chronostratigraphic constraints allow tentative correlations to be made between Profitis Ilias and Pindos-Olonos sedimentary units. Finally, the palaeoceanographic significance of the studied series in Rhodes and potentially similar pelagic sequences in the Marmaris area of Turkey are discussed

Methamphetamine Induces TET1- and TET3-Dependent DNA Hydroxymethylation of Crh and Avp Genes in the Rat Nucleus Accumbens

Methamphetamine (METH) addiction is a biopsychosocial disorder that is accompanied by multiple relapses even after prolonged abstinence, suggesting the possibilities of long-lasting maladaptive epigenetic changes in the brain. Here, we show that METH administration produced time-dependent increases in the expression of corticotropin-releasing hormone (Crh/Crf), arginine vasopressin (Avp), and cocaine- and amphetamine-regulated transcript prepropeptide (Cartpt) mRNAs in the rat nucleus accumbens (NAc). Chromatin immunoprecipitation (ChIP) assays revealed that METH increased the abundance of phosphorylated CREB (pCREB) at the promoter of Cartpt but not at Avp or Crh DNA sequences. In contrast, METH produced DNA hypomethylation at sites near the Crh transcription start site (TSS) and at intragenic Avp sequences. METH also increased DNA hydroxymethylation at the Crh TSS and at intragenic Avp sites. In addition, METH increased the protein expression of ten-eleven-translocation enzymes that catalyze DNA hydroxymethylation. Importantly, METH increased TET1 binding at the Crh promoter and increased TET3 binding at Avp intragenic regions. We further tested the role of TET enzymes in METH-induced changes in gene expression by using the TET inhibitor, 1,5-isoquinolinediol (IQD), and found that IQD blocked METH-induced increases in Crh and Avp mRNA expression. Together, these results indicate that METH produced changes in neuropeptide transcription by both activation of the cAMP/CREB pathway and stimulation of TET-dependent DNA hydroxymethylation. These results provide molecular evidence for epigenetic controls of METH-induced changes in the expression of neuropeptides.Fil: Jayanthi, Subramaniam. National Institutes of Health; Estados UnidosFil: Gonzalez, Betina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; ArgentinaFil: McCoy, Michael T.. National Institutes of Health; Estados UnidosFil: Ladenheim, Bruce. National Institutes of Health; Estados UnidosFil: Bisagno, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; ArgentinaFil: Cadet, Jean Lud. National Institutes of Health; Estados Unido