Concurrent beneficial (vitamin D production) and hazardous (cutaneous DNA damage) impact of repeated low-level summer sunlight exposures.

BACKGROUND: The concurrent impact of repeated low-level summer sunlight exposures on vitamin D production and cutaneous DNA damage, potentially leading to mutagenesis and skin cancer, is unknown. OBJECTIVES: This is an experimental study (i) to determine the dual impact of repeated low-level sunlight exposures on vitamin D status and DNA damage/repair (via both skin and urinary biomarkers) in light-skinned adults; and (ii) to compare outcomes following the same exposures in brown-skinned adults. METHODS: Ten white (phototype II) and six South Asian volunteers (phototype V), aged 23-59 years, received 6 weeks' simulated summer sunlight exposures (95% ultraviolet A/5% ultraviolet B, 1·3 standard erythemal doses three times weekly) wearing summer clothing exposing ~35% body surface area. Assessments made were circulating 25-hydroxyvitamin D [25(OH)D], immunohistochemistry for cyclobutane pyrimidine dimer (CPD)-positive nuclei and urinary biomarkers of direct and oxidative (8-oxo-deoxyguanosine) DNA damage. RESULTS: Serum 25(OH)D rose from mean 36·5 ± 13·0 to 54·3 ± 10·5 nmol L-1 (14·6 ± 5·2 to 21·7 ± 4·2 ng mL-1 ) in phototype II vs. 17·2 ± 6·3 to 25·5 ± 9·5 nmol L-1 (6·9 ± 2·5 to 10·2 ± 3·8 ng mL-1 ) in phototype V (P < 0·05). Phototype II skin showed CPD-positive nuclei immediately postcourse, mean 44% (range 27-84) cleared after 24 h, contrasting with minimal DNA damage and full clearance in phototype V (P < 0·001). The findings did not differ from those following single ultraviolet radiation (UVR) exposure. Urinary CPDs remained below the detection threshold in both groups; 8-oxo-deoxyguanosine was higher in phototype II than V (P = 0·002), but was unaffected by UVR. CONCLUSIONS: Low-dose summer sunlight exposures confer vitamin D sufficiency in light-skinned people concurrently with low-level, nonaccumulating DNA damage. The same exposures produce minimal DNA damage but less vitamin D in brown-skinned people. This informs tailoring of sun-exposure policies.

Does nausea and vomiting of pregnancy play a role in the association found between maternal caffeine intake and fetal growth restriction?

The aim of this study was to explore the relationships between nausea and vomiting in pregnancy and (a) fetal growth restriction; and (b) maternal caffeine metabolism and fetal growth restriction. A cohort of 2,643 pregnant women, aged 18-45 years, attending two UK maternity units between 8 and 12 weeks gestation, was recruited. A validated tool assessed caffeine intake at different stages of pregnancy and caffeine metabolism was assessed from a caffeine challenge test. Experience of nausea and vomiting of pregnancy was self-reported for each trimester. Adjustment was made for confounders, including salivary cotinine as a biomarker of current smoking status. There were no significant associations between fetal growth restriction and nausea and vomiting in pregnancy, even after adjustment for smoking and alcohol intake. There were no significant differences in the relationship between caffeine intake and fetal growth restriction between those experiencing symptoms of nausea and vomiting and those who did not, for either the first (p = 0.50) or second trimester (p = 0.61) after adjustment for smoking, alcohol intake and caffeine half-life. There were also no significant differences in the relationship between caffeine half-life and fetal growth restriction between those experiencing symptoms of nausea and vomiting and those who did not, for either the first trimester (p = 0.91) or the second trimester (p = 0.45) after adjusting for smoking, alcohol intake and caffeine intake. The results from this study show no evidence that the relationship between maternal caffeine intake and fetal growth restriction is modified by nausea and vomiting in pregnancy.

First-trimester increase in oxidative stress and risk of small-for-gestational-age fetus.

OBJECTIVE: Investigation of increased oxidative stress in early pregnancy and association with an increased risk of small-for-gestational-age (SGA) fetus. DESIGN: Longitudinal case-control study. SETTING: University Hospitals of Leicester NHS Trust, Leicester, UK. POPULATION: Low-risk pregnant women with no current or pre-existing medical illness were recruited at a large teaching hospital from 2004 to 2006. METHODS: Recruitment performed at the time of the dating ultrasound scan (12+/-2 weeks of gestation). Spot urine samples collected at 12+/-2 and 28+/-2 weeks of gestation were analysed for 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) by liquid chromatography with tandem mass spectrometry). SGA was defined as birthweight <10th centile based on customised centile calculator (www.gestation.net). This identified the cases (n=55), whereas controls (n=55) were mothers whose babies were appropriate for gestational age (AGA, birthweight 10th-90th centile). Statistical analysis was performed using GraphPad Prism v.5. The relationship between maternal urinary 8-oxodG at different gestations and customised SGA was investigated by nonparametric tests. MAIN OUTCOME MEASURES: Customised SGA and AGA pregnancies. RESULTS: Urinary 8-oxodG concentrations were significantly increased in pregnancies with subsequent SGA compared with concentrations in normal pregnancies; 12 weeks: 2.8 (interquartile range [IQR] 1.96-3.67) versus 2.2 (IQR 1.26-3.28) pmol 8-oxodG/micromol creatinine (P=0.0007); 28 weeks: 2.21 (IQR 1.67-3.14) versus 1.68 (IQR 1.16-2.82) pmol 8-oxodG/micromol creatinine (P<0.0002). Concentrations decreased significantly between week 12 and 28 (P=0.04 and P=0.02 for controls and cases). CONCLUSIONS: In this study, urinary 8-oxodG at 12 and 28 weeks were elevated in SGA compared with AGA pregnancies. This may reflect early placental changes predating clinical features of SGA.

Oxidative damage to DNA in non-malignant disease: biomarker or biohazard?

Oxidative damage to DNA has been examined in many non-malignant conditions, in most cases for its utility as a marker of oxidative stress. Whilst this may prove useful, attempts to answer the question - why might oxidative damage be important in this disease? - would provide added value to the biomarker data, as well as give clues to pathogenesis and perhaps therapy. In this chapter, data from the scientific literature are considered broadly, where oxidative damage to DNA has been analysed either in tissues or in extracellular matrices, such as urine, in various groups of non-malignant disease. The lesion of primary focus is 8-hydroxy-7,8-dihydro-2'-deoxyguanosine, only because this is the most widely measured lesion. By coupling biomarker information with the characteristics of the disease and a set of general mechanisms whereby DNA oxidation may be pathogenic (retrospectively derived from the literature examined), we can ascribe pathogenic roles for DNA oxidation in various diseases. Based on available experimental evidence, for a wide range of conditions, such mechanisms would include prominent roles for the induction of mitochondrial dysfunction, promotion of cytotoxicity and modulation of inflammatory responses. Our general conclusion is that, dependent on the disease, oxidative DNA damage may be a biomarker, biohazard or both of these.

Comparing the attitudes of urban and rural Iranian women toward menopause.

OBJECTIVES: This study was designed to compare the attitudes of urban and rural Iranian women toward menopause. The socio-cultural context of urban Tehranian women is considerably more westernized than that of rural Semiromian women. Our original hypothesis was that urban Tehranian women would have a more negative outlook on menopause than rural women. This hypothesis was drawn from previous work implicating the youth-oriented culture of the west as responsible for the negative attitude of western women toward menopause. METHODS: A questionnaire with eight questions regarding major psychosocial issues that are of importance to menopausal women was prepared and administered to a group of 70 Tehranian and 49 Semiromian women. RESULTS: In contrast to the initial hypothesis, analysis revealed that rural Iranian women have a more negative attitude towards menopause than urban Iranian women. CONCLUSION: The more negative attitude of the rural women toward menopause in large part reflected the higher priority they placed on fertility than did their urban counterparts. Further, cross-cultural studies will illuminate exactly which social and cultural factors play a role in both the physical and psychological aspects of menopause.

Role of dietary antioxidants in the prevention of in vivo oxidative DNA damage.

Epidemiological evidence consistently shows that diets high in fresh fruit and vegetables significantly lower cancer risk. Given the postulated role of oxidative DNA damage in carcinogenesis, the assumption has been made that it is the antioxidant properties of food constituents, such as vitamin C, E and carotenoids, which confer protection. However, epidemiological studies with specific antioxidants, either singly or in combination, have not, on the whole, supported this hypothesis. In contrast, studies examining the in vitro effect of antioxidants upon oxidative DNA damage have generally been supportive, in terms of preventing damage induction. The same, however, cannot be said for the in vivo intervention studies where overall the results have been equivocal. Nevertheless, recent work has suggested that some dietary antioxidants may confer protective properties through a novel mechanism, unrelated to their conventional free-radical scavenging abilities. Upregulation of antioxidant defence, xenobiotic metabolism, or DNA-repair genes may all limit cellular damage and hence promote maintenance of cell integrity. However, until further work has clarified whether dietary supplementation with antioxidants confers a reduced risk of cancer and the mechanism by which this effect is exerted, the recommendation for a diet rich in fruit and vegetables remains valid empirically.

Monoclonal antibody to single-stranded DNA: a potential tool for DNA repair studies.

Growing evidence suggests that DNA repair capacity is an important factor in cancer risk and is therefore essential to assess. Immunochemical assays are amenable to the detection of repair products in complex matrices, such as urine, facilitating noninvasive measurements, although diet and extra-DNA sources of lesion can confound interpretation. The production of single-stranded, lesion-containing DNA oligomers characterises nucleotide excision repair (NER) and hence defines the repair pathway from which a lesion may be derived. Herein we describe the characterisation of a monoclonal antibody which recognises guanine moieties in single-stranded DNA. Application of this antibody in ELISA, demonstrated such oligomers in supernatants from repair-proficient cells post-insult. Testing of urine samples from volunteers demonstrated a relationship between oligomer levels and two urinary DNA damage products, thymine dimers and 8-oxo-2'-deoxyguanosine, supporting our hypothesis that NER gives rise to lesion-containing oligomers which are specific targets for the investigation of DNA repair.

Induction and excretion of ultraviolet-induced 8-oxo-2'-deoxyguanosine and thymine dimers in vivo: implications for PUVA.

Molecular epidemiology has linked ultraviolet-induced DNA damage with mutagenesis and skin carcinogenesis. Ultraviolet radiation may damage DNA in one of two ways: either directly, leading to lesions such as cyclobutane thymine dimers (T<>T), or indirectly, via photosensitizers that generate free radical species that may ultimately produce such oxidative lesions as 8-oxo-2'-deoxyguanosine. We report the results of a pilot, case control study in which seven, healthy, human volunteers (skin type II; aged 23-56 y; three male, four female) received a suberythemal dose of whole body irradiation from ultraviolet-A-emitting fluorescent tubes used in psoralen plus ultraviolet A therapy. First void, mid-stream urine samples were collected pre-exposure and daily postexposure, for up to 13 d. Analysis of urinary 8-oxo-2'-deoxyguanosine and cyclobutane thymine dimers was by competitive enzyme-linked immunosorbent assay (interassay coefficient of variation < or = 10%) and compared with a matched, control group of unirradiated individuals. A maximal increase in levels of urinary 8-oxo-2'-deoxyguanosine was seen 4 d post-ultraviolet exposure. A subsequent reduction was noted, before finally returning to baseline. Similarly, cyclobutane thymine dimer levels peaked 3 d postexposure, before returning to baseline. In contrast to the 8-oxo-2'-deoxyguanosine analysis, however, a second peak was noted at days 9-11, before again returning to baseline. This is the first report examining urinary 8-oxo-2'-deoxyguanosine and cyclobutane thymine dimers following ultraviolet exposure of healthy human subjects. This work illustrates the induction and time course for excretion of ultraviolet-induced lesions, perhaps alluding to repair and ultimately offering the potential to define psoralen plus ultraviolet A dosage regimes in terms of minimizing DNA damage and hence cancer risk.

Laboratory evaluation of mouthguard material.

The aim of this study was to evaluate, in vitro, the EVA material (polyvinylacetate-polyethylene copolymer) most commonly used in the fabrication of mouthguards and thus to help understand mouthguard performance and to improve mouthguard design. The material was tested for tensile strength, elongation, hardness and water absorption using specimens and tests described by the American Society for Testing and Materials Standards. Standard EVA sheets of 1, 2, 3 and 5 mm thickness were prepared. With increase in the thickness of the material, both the ultimate tensile strength and the elongation were decreased. The hardness and water absorption were unaffected. It was concluded that. overall, the thicker 5 mm EVA material was recommended for mouthguards as it displayed the least deformation to load and performed equally as well in the tests as other thicknesses.

The role of the mouthguard in the prevention of sports-related dental injuries: a review.

OBJECTIVES: This paper examines the literature dealing with oral-facial injuries received during participation in sport and the possibilities open to athletes for their prevention. In particular, the paper examines five different aspects of this topic: the risk of dental injury while playing sports, the role of the mouthguard in preventing injury, types of athletic mouthguard, implications for patients undergoing orthodontic treatment and behavioural aspects of mouthguard wear. RESULTS: It is clear from this review that participation in a number of sports does carry a considerable risk of sustaining dental injury, not only in the so-called contact sports such as rugby and hockey, but also in less obviously dangerous sports such as basketball. Although some evidence exists to the contrary, the majority of studies have found the mouthguard to be the most effective way of preventing such injuries. It is also clear that the custom-fabricated mouthguard, in particular the pressure-laminated variety, is seen to afford most protection. Athletes undergoing orthodontic treatment present a particular problem as they are potentially at greater risk of injury because of increased tooth mobility and the presence of orthodontic appliances. The fabrication of mouthguards for these patients is also problematic and the literature covering this is reviewed. As with other preventive measures, mouthguard usage is often less than the dental profession would like; the reasons for this are explored in a small number of studies. CONCLUSION: While much progress has been made in this area, the profession could do much more to promote the greater use of mouthguards.

Immunochemical quantitation of UV-induced oxidative and dimeric DNA damage to human keratinocytes.

There is growing evidence to suggest that solar radiation-induced, oxidative DNA damage may play an important role in skin carcinogenesis. Numerous methods have been developed to sensitively quantitate 8-oxo-2'deoxyguanosine (8-oxodG), a recognised biomarker of oxidative DNA damage. Immunoassays may represent a means by which the limitations of many techniques, principally derived from DNA extraction and sample workup, may be overcome. We report the evaluation of probes to thymine dimers and oxidative damage in UV-irradiated cells and the DNA derived therefrom. Thymine dimers were most readily recognised, irrespective of whether in situ in cells or in extracted DNA. However, using antibody-based detection the more subtle oxidative modifications required extraction and, in the case of 8-oxodG, denaturation of the DNA prior to successful recognition. In contrast, a recently described novel probe for 8-oxodG detection showed strong recognition in cells, although appearing unsuitable for use with extracted DNA. The probes were subsequently applied to examine the relative induction of lesions in cells following UV irradiation. Guanine-glyoxal lesions predominated over thymine dimers subsequent to UVB irradiation, whereas whilst oxidative lesions increased significantly following UVA irradiation, no induction of thymine dimers was seen. These data support the emerging importance of oxidative DNA damage in UV-induced carcinogenesis.

Aberrant processing of oxidative DNA damage in systemic lupus erythematosus.

Defective DNA damage processing has been reported in systemic lupus erythematosus (SLE). Vitamin C may modulate formation/removal of the oxidative DNA lesion 8-oxo-2'-deoxyguanosine (8-oxodG). Baseline levels of 8-oxodG measured in SLE serum, urine and PBMC DNA did not differ significantly from healthy subjects. In contrast to healthy subjects, no significant decrease in PBMC 8-oxodG or increase in urinary 8-oxodG was noted in vitamin C supplemented SLE patients. A significant, although attenuated, increase in serum 8-oxodG was detected in SLE patients, compared to healthy subjects. These data support putative abnormalities in the repair/processing of 8-oxodG in SLE.

Urinary 8-oxo-2'-deoxyguanosine--source, significance and supplements.

Oxidative damage to cellular biomolecules, in particular DNA, has been proposed to play an important role in a number of pathological conditions, including carcinogenesis. A much studied consequence of oxygen-centred radical damage to DNA is 8-oxo-2'-deoxyguanosine (8-oxodG). Using numerous techniques, this lesion has been quantified in various biological matrices, most notably DNA and urine. Until recently, it was understood that urinary 8-oxodG derives solely from DNA repair, although the processes which may yield the modified deoxynucleoside have never been thoroughly discussed. This review suggests that nucleotide excision repair and the action of a specific endonuclease may, in addition to the nucleotide pool, contribute significantly to levels of 8-oxodG in the urine. On this basis, urinary 8-oxodG represents an important biomarker of generalised, cellular oxidative stress. Current data from antioxidant supplementation trials are examined and the potential for such compounds to modulate DNA repair is considered. It is stressed that further work is required to link DNA, serum and urinary levels of 8-oxodG such that the kinetics of formation and clearance may be elucidated, facilitating greater understanding of the role played by oxidative stress in disease.

Urinary thymine dimers and 8-oxo-2'-deoxyguanosine in psoriasis.

Psoralen in conjunction with UVA (PUVA) is perhaps the most effective treatment for psoriasis. It is, however, a risk factor for skin cancer in these patients and there is a need to develop non-invasive assays reflective of treatment-induced DNA damage. We report here the assessment of two important lesions, thymine dimer (T<>T) and 8-oxo-2'-deoxyguanosine (8-OHdG), in the urine of psoriasis patients. It was found that, once corrected for urine concentration, the psoriatic group had significantly higher (P<0. 0001) urinary levels of thymine dimers compared to the control group. No significant differences in urinary 8-OHdG levels were noted between the psoriatic, atopic dermatitis and control groups. Therefore biomonitoring of therapy from the very start with this simple and non-invasive assay could perhaps be an effective measure of the risk involved with the treatment allowing optimization for minimal-risk therapy.

Fifteen-year reproducibility of natural head posture: A longitudinal study.

Natural head posture continues to be widely used as the logical reference position for the evaluation of craniofacial morphology. The basic underlying premise is that the long-term clinical reproducibility (variability) of natural head posture is significantly less than the variability of conventional reference planes with respect to the vertical. This study reports the 15-year longitudinal reproducibility of natural head posture. Twenty Chinese adults in Hong Kong, who had initial natural head posture radiographs at age 12 years, were followed up and had repeated cephalograms after 15 years. The method error (reproducibility) after 15 years was 2.2 degrees, which compared favorably with the 5-year reproducibility (method error = 3.0 degrees ) and the 5 to 10 minutes reproducibility (method error = 1.9 degrees ). The individual variability of natural head posture reproducibility increased slightly over time. After 15 years the variance of natural head posture (4.8 degrees [= 2.2(2)]) remains significantly less than the variance of intracranial reference planes to the vertical (25 degrees to 36 degrees ). Cephalometric analyses based on natural head posture therefore remain valid over time.

Discrepancies in the measurement of UVC-induced 8-oxo-2'-deoxyguanosine: implications for the analysis of oxidative DNA damage.

Ultraviolet (UV) light-induced indirect, oxidative damage to DNA has received increasing attention with respect to the mutagenic and carcinogenic effects of solar radiation. An oxidative lesion that has raised particular interest because of its qualitative and quantitative importance is 8-oxo-2'-deoxyguanosine. This deoxynucleoside lesion is most frequently measured by high performance liquid chromatography with electrochemical detection (HPLC-EC) following enzymatic hydrolysis of DNA or as the base equivalent, 8-oxoguanine, by gas chromatography-mass spectrometry (GC-MS) following acid hydrolysis of DNA. We have noted a discrepancy in the literature whereby the levels of 8-oxo-2'-deoxyguanosine measured by HPLC-EC in UVC-irradiated DNA are significantly higher than when 8-oxoguanine is measured by GC-MS. By making use of the availability of both HPLC-EC and stable-isotope dilution GC-MS methodologies in our laboratory we have confirmed the discrepancy noted in the literature by parallel analysis of the same UVC-irradiated calf thymus DNA samples. Furthermore, analysis of the UVC-induced product by UV-visible spectrophotometry, voltammetry and its detection by a monoclonal antibody which recognises 8-oxo-2'-deoxyguanosine strongly suggests that the product is indeed 8-oxo-2'-deoxyguanosine. Partial explanation for this discrepancy could be an inordinate resistance of UVC-irradiated DNA to formic acid hydrolysis. However, we cannot completely exclude the possibility that there is a formic acid-labile species which co-elutes with 8-oxo-2'-deoxyguanosine in enzymatically digested UVC-irradiated DNA. Whether this phenomenon is unique to UV-irradiation damage or occurs with other systems that cause oxidative damage to DNA awaits further investigation. Irrespective of the exact mechanism, there will be significant implications for the analysis of oxidative DNA damage.

Comparison of cephalometric analysis using a non-radiographic sonic digitizer (DigiGraph Workstation) with conventional radiography.

Cephalometric analysis conventionally requires radiographic exposure which may not be compatible with the growing concern over radiation hazards. Recently, the Dolphin Workstation Imaging System introduced to the dental profession a non-radiographic system, called the DigiGraph Workstation which may be an alternative to cephalometric radiography. The aims of this study were to compare the validity and reproducibility of cephalometric measurements obtained from the DigiGraph Workstation with conventional cephalometric radiographs. The sample consisted of 30 human dry skulls. Two replicated sets of lateral cephalograms were obtained with steel ball markers placed at the majority of the cephalometric landmarks. Duplicate tracings prepared from each radiograph were digitized to obtain cephalometric measurements using the computer software, Dentofacial Planner. For the DigiGraph Workstation, double sonic digitizations were repeated twice for each skull, on two occasions. Fifteen angular and one linear measurements were obtained from both methods and these findings compared using ANOVA, paired t-tests and F-tests. All, except one, cephalometric measurement showed significant differences between the two methods (P < 0.0001). The DigiGraph Workstation consistently produced higher values in 11 measurements (mean differences +0.5 to +15.7 degrees or mm) and lower values in four measurements (mean differences -0.2 to -3.5 degrees). The standard deviations of the differences between readings of both methods were large (0.4-5.8 degrees or mm). The reproducibility of the DigiGraph Workstation measurements was lower than that of the radiographic measurements. The method error of the DigiGraph Workstation ranged from 7 to 70 per cent, while that of radiographic tracings was less than 2 per cent. It was concluded that measurements obtained with the DigiGraph Workstation should be interpreted with caution.