Recovery of DNA from fired and unfired cartridge casings: comparison of two DNA collection methods.

For over 10 years, various studies have attempted to increase the recovery of DNA from ammunition by modifying the DNA collection, extraction, purification, and amplification procedures, with varying levels of success. This study focused on the "soaking" method of Montpetit & O'Donnell [1] and the "rinse-and-swab" method of Bille et al. [2]. First, testing for the presence of exogenous DNA, 210 boxed cartridges (brass, steel, and nickel-plated) from nine manufacturers were swabbed and DNA was extracted, concentrated, and quantified. Extracts that quantified > 0 ng/µL (44 of 210) were amplified and genotyped with GlobalFiler™. Of those, only one extract yielded two alleles indicating that the manufacturing and packaging of ammunition was virtually DNA free. Next, to obtain a baseline comparison of two DNA collection methods on a non-metallic substrate and identify a suitable number of cells to spot on cartridges, different DNA input amounts of primary human adult epidermal keratinocytes (HEKa) were tested. Thereafter, 300 brass and 300 nickel-plated, cartridges were spotted with HEKa cells containing ~5 ng of DNA, fired or unfired, and processed with either method. Finally, five methods representing hybrids of the soaking and rinse-and-swab methods were tested to determine if variations of those methods could be used to increase DNA yield and recovery. The results show that the soaking method consistently yielded more DNA than the rinse-and-swab method from a non-metallic substrate. However, the comparison study demonstrated that both methods performed comparably for cartridges. On average, the soaking method recovered 0.25 ng of DNA (5.1% recovery) and the rinse-and-swab method recovered 0.28 ng (5.8% recovery). However, average recoveries were significantly different among three analysts and considerable variation in yields were observed, possibly due to storage time. Furthermore, consistent with prior reports, the DNA recovered from brass casings was only 16% of that recovered from nickel-plated casings and the average yield of DNA from fired casings was reduced to 67% of unfired casings. Moreover, DNA extracts from brass or nickel-plated casings did not appear to contain amplification inhibitors and only 30/596 appeared severely degraded. Finally, both the published rinse-and-swab and soaking methods yielded more DNA than all modifications of the two methods. Overall, both methods yielded equivalent DNA quantities. Additionally, recovery of DNA from any given cartridge casing may be dependent on storage time as well as the skill, proficiency, and experience of the analyst and may reflect stochastic effects, particularly for casings containing low copy and/or degraded DNA.

Life History and Damage by Systena frontalis F. (Coleoptera: Chrysomelidae) on Vaccinium macrocarpon Ait.

Pest management of emerging pests can be challenging because very little fundamental knowledge is available to inform management strategies. One such pest, the red-headed flea beetle Systena frontalis (Fabricius) (Coleoptera: Chrysomelidae), is increasingly being identified as a pest of concern in cranberries Vaccinium macrocarpon Aiton (Ericales: Ericaceae). To improve our understanding of this pest and to develop more targeted management programs, we conducted field and laboratory studies to characterize the development, seasonal emergence patterns, and density-dependent plant injury. We found that significantly more flea beetle eggs hatched when exposed to sustained cold treatment between 0 and 5°C for 15 wk than at warmer temperatures, and for shorter and longer cold-period durations. The adults emerged sporadically over the summer, were patchily distributed, fed on both fruit and foliage, and preferentially fed on new plant growth. Using soil cores, we found eggs and larvae located relatively deep (>30 cm) in the soil. These patterns indicate that S. frontalis likely overwinters as eggs, and that targeting the larval stage may be the most effective management approach. Despite the cryptic nature of the larvae, continuing to improve our understanding of this life stage will be critical to optimizing control strategies.

Associations between hair manganese levels and cognitive, language, and motor development in preschool children from Montevideo, Uruguay.

Environmental manganese (Mn) exposure in childhood has been associated with developmental deficits. This study investigated associations between low-level Mn exposure and developmental scores in children aged 14-45 months. Children's hair samples (n = 60) were analyzed for Mn and lead (Pb) using inductively coupled plasma mass spectrometry (ICP-MS). Children were assessed with the Bayley Scales of Infant Development III (BSID). Mean hair Mn and Pb concentrations were 0.98 ± 0.74 and 10.1 ± 10.5 µg/g, respectively. Unadjusted inverse associations were seen between Mn and BSID subscales, with girls having lower scores related to Mn than boys. No significant associations remained in covariate-adjusted models, although when stratified by sex boys had a positive association between hair Mn levels and language scores. This study does not show clear evidence of an association between hair Mn concentrations and children's developmental scores in a region of low-level Mn exposure.

Self-report of fruit and vegetable intake that meets the 5 a day recommendation is associated with reduced levels of oxidative stress biomarkers and increased levels of antioxidant defense in premenopausal women.

BACKGROUND: Oxidative stress has been associated with a variety of chronic diseases and reproductive disorders. Fruits and vegetables (F/V) may contribute to antioxidant vitamin and micronutrient levels and reduce oxidative stress. OBJECTIVE: To investigate the effect of meeting the 5 A Day For Better Health Program recommendation for F/V consumption on biomarkers of oxidative damage and antioxidant defense. DESIGN: In this longitudinal study, healthy premenopausal women (n=258) were followed for ≤2 menstrual cycles with ≤16 oxidative stress measures timed to cycle phase. MAIN OUTCOME MEASURES: Plasma concentrations of F2-isoprostane, 9-hydroxyoctadecadieneoic acid, 13-hydroxyoctadecadieneoic acid, erythrocyte activity of superoxide dismutase, glutathione reductase, and glutathione peroxidase, as well as blood micronutrient concentrations were measured. Dietary intake was assessed by food frequency questionnaires (FFQs) (1 per cycle), and 24-hour recalls (≤4 per cycle). STATISTICAL ANALYSES PERFORMED: Fruit and vegetable servings were dichotomized based on the recommendation to consume five servings of F/V each day. Linear mixed models with repeated measures were used to analyze lipid peroxidation markers, antioxidant vitamins, and antioxidant enzymes by cycle phase and in association with usual F/V intake. RESULTS: For both 24-hour recall (timed to cycle phase) and cycle-specific FFQ, meeting the recommendation to consume five servings of F/V each day was associated with decreased F2-isoprostanes (24-hour recall ß=-.10 [95% CI, -0.12 to -0.07]; FFQ ß= -.14 [95% CI, -0.18 to -0.11]). Glutathione reductase was lower in association with typical consumption of five or more servings of F/V by FFQ but not in the phase-specific analysis. Higher levels of ascorbic acid, lutein, beta carotene, and beta cryptoxanthin were observed with both intake measures. CONCLUSIONS: Meeting the 5 A Day For Better Health Program recommendation was associated with lower oxidative stress and improved antioxidant status in analyses of typical diet (via FFQ) and in menstrual cycle phase-specific analyses using 24-hour recalls. Green salads were commonly eaten and increasing intake of salads may be a useful strategy to influence oxidation in reproductive aged women.

pH effect on the susceptibility to parasitoid infection in the marine diatom Coscinodiscus spp. (Bacillariophyceae).

The pH on the frustule of individual cells of the marine centric diatoms Coscinodiscus granii and Coscinodiscus wailesii (Bacillariophyceae) was measured with pH microsensors in culture media with increasing pH values of 8.04, 8.14, and 8.22, respectively. In 85-96% of the C.granii cells the pH on the frustule was up to 0.4 units higher than that of the medium, reaching a maximum pH 8.95. Only in 2-3% the surface pH exceeded that of the medium by up to 0.7 pH units. These results strongly suggest that diatoms in batch cultures differ, at least temporarily, in their individual photosynthetic activities. Infection experiments with the parasitoid nanoflagellate Pirsonia diadema (Stramenopile) showed that flagellates failed to infect when the culture pH was 8.8 and above. pH measurements on freshly infected C. granii showed that the prevalence of infection was higher in tendency on diatoms with low surface pH. Application of these results to parasitoid-diatom interactions in natural waters suggests that within phytoplankton populations a strong photosynthetic activity might prevent diatom cells temporarily from infection by pH-sensitive parasitoids.