Hyperbaric oxygen preconditioning protects skin from UV-A damage

Hyperbaric oxygen therapy (HBOT) is used for a number of applications, including the treatment of diabetic foot ulcers and CO poisoning. However, we and others have shown that HBOT can mobilize cellular antioxidant defenses, suggesting that it may also be useful under circumstances in which tissue protection from oxidative damage is desired. To test the protective properties of hyperbaric oxygen (HBO) on a tissue level, we evaluated the ability of a preconditioning treatment regimen to protect cutaneous tissue from UV-A-induced oxidative damage. Three groups of hairless SKH1-E mice were exposed to UV-A 3 days per week for 22 weeks, with two of these groups receiving an HBO pretreatment either two or four times per week. UV-A exposure increased apoptosis and proliferation of the skin tissue, indicating elevated levels of epithelial damage and repair. Pretreatment with HBO significantly reduced UV-A-induced apoptosis and proliferation. A morphometric analysis of microscopic tissue folds also showed a significant increase in skin creasing following UV-A exposure, which was prevented by HBO pretreatment. Likewise, skin elasticity was found to be greatest in the group treated with HBO four times per week. The effects of HBO were also apparent systemically as reductions in caspase-3 activity and expression were observed in the liver. Our findings support a protective function of HBO pretreatment from a direct oxidative challenge of UV-A to skin tissue. Similar protection of other tissues may likewise be achievable.     

Uncertainty and Variability of Energy and Material Use by Fused Deposition Modeling Printers in Makerspaces

Desktop‐grade fused deposition modeling (FDM) printers are popular because of compact sizes and affordable prices. If we are moving toward a future where desktop FDM printers are in every school and office, like conventional printers, then these machines will consume a large amount of energy and material. However, it is very difficult to evaluate the environmental impacts of FDM printers since there are so many different brands and types of printers using different raw materials under different scenarios. This study uses data from two different printing sites to evaluate the scenario and parameter uncertainty and variability in energy and material balances for FDM printers. Data from the two makerspaces provide insight into the material and energy consumption data using polylactic acid and acrylonitrile butadiene styrene (ABS) with four types of printers. The use of actual performance data allowed for the additional study of scrap ratio. Regressions provide insight into predictive factors for energy and material consumption. Monte Carlo simulations show the range of energy life cycle inventory values for the desktop‐grade FDM printers. From the regressions, Type A Pro was the most energy‐intensive machine. For material waste, an open‐access makerspace using ABS was associated with higher scrap ratio. Regression analysis indicates that the rate of material usage is not a strong predictor of waste rates. The amount of waste generated across both sites indicates that more ubiquitous access to FDM printing may create a significant addition to the waste stream.     

Engineered genetic selection links in vivo protein folding and stability with asparagine-linked glycosylation

Predicting the structural consequences of site-specific glycosylation remains a major challenge due in part to the lack of convenient experimental tools for rapidly determining how glycosylation influences protein folding. To address this shortcoming, we developed a genetic selection that directly links the in vivo folding of asparagine-linked (N-linked) glycoproteins with antibiotic resistance. Using this assay, we identified three known or putative glycoproteins from Campylobacter jejuni (Peb3, CjaA, and Cj0610c) whose folding was significantly affected by N-glycosylation. We also used the genetic selection to isolate a glycoengineered variant of the Escherichia coli colicin E7 immunity protein (Im7) whose intracellular folding and stability were enhanced as a result of N-glycosylation. In addition to monitoring the effect of glycan attachment on protein folding in living cells, this strategy could easily be extended for optimizing protein folding in vivo and engineering glycosylation enzymes, pathways, and hosts for optimal performance. See accompanying commentary by Danielle Tullman-Ercek DOI: 10.1002/biot.201300319     

Prostaglandins temporally regulate cytoplasmic actin bundle formation during Drosophila oogenesis

Prostaglandins (PGs)—lipid signals produced downstream of cyclooxygenase (COX) enzymes—regulate actin dynamics in cell culture and platelets, but their roles during development are largely unknown. Here we define a new role for Pxt, the Drosophila COX-like enzyme, in regulating the actin cytoskeleton—temporal restriction of actin remodeling during oogenesis. PGs are required for actin filament bundle formation during stage 10B (S10B). In addition, loss of Pxt results in extensive early actin remodeling, including actin filaments and aggregates, within the posterior nurse cells of S9 follicles; wild-type follicles exhibit similar structures at a low frequency. Hu li tai shao (Hts-RC) and Villin (Quail), an actin bundler, localize to all early actin structures, whereas Enabled (Ena), an actin elongation factor, preferentially localizes to those in pxt mutants. Reduced Ena levels strongly suppress early actin remodeling in pxt mutants. Furthermore, loss of Pxt results in reduced Ena localization to the sites of bundle formation during S10B. Together these data lead to a model in which PGs temporally regulate actin remodeling during Drosophila oogenesis by controlling Ena localization/activity, such that in S9, PG signaling inhibits, whereas at S10B, it promotes Ena-dependent actin remodeling.     

Comparison of protein quantification and extraction methods suitable for E. coli cultures

Many different extraction and analysis methods exist to determine the protein fraction of microbial cells. For metabolic engineering purposes it is important to have precise and accurate measurements. Therefore six different protein extraction protocols and seven protein quantification methods were tested and compared. Comparison was based on the reliability of the methods and boxplots of the normalized residuals. Some extraction techniques (SDS/chloroform and toluene) should never be used: the measurements are neither precise nor accurate. Bugbuster extraction combined with UV280 quantification gives the best results, followed by the combinations Sonication-UV280 and EasyLyse-UV280. However, if one does not want to use the quantification method UV280, one can opt to use Bugbuster, EasyLyse or sonication extraction combined with any quantification method with exception of the EasyLyse-BCA_P and Sonication-BCA_P combinations.     

Tadpole fingerprinting: Using tail venation patterns to photo-identify tadpole individuals of a threatened frog

Traditional methods for identifying individual amphibians in capture–mark–recapture (CMR) studies have been primarily confined to post-metamorphic stages, using artificial markers that come with a variety of limitations. An alternative that may open CMR studies to earlier life stages involves the use of a species' natural external markers in photo-based identification. In this study, we investigated whether it was possible to distinguish tadpoles of the threatened green and golden bell frog (Litoria aurea) at the individual level based on tail venation patterns. We collected photographs of the tails of captive-raised tadpoles using a smartphone over a 4-week period. This photo-library was used to create an electronic survey where participants were asked to detect matches for query tadpoles from small image pools. We found that most participants agreed on a match for each query, with perfect consensus achieved for most queries (83%). We detected a 14% decline in perfect consensus when participants were asked to match images of tadpoles separated by longer time intervals, suggesting that it is more difficult to visually identify recapture events of L. aurea tadpoles over extended periods due to changes to tail appearance. However, consensus was obtained by participants for all queries, with all matches verified as being correct by the primary researcher. The strength of agreement among participants with no prior experience in matching tadpole tails suggests that there is sufficient inter-individual variation in this feature for individuals to be manually identified. We thus propose that photo-identification is likely to be a valid, non-invasive technique that can be used for short-term studies on tadpole populations that display tail venation. This offers an alternative to artificial markers that may not allow for individual identification, while also opening up tadpole monitoring programmes to citizen scientists who can be recruited online to process image data from home.     

Prairie Revegetation of a Strip Mine in Illinois: Fifteen Years after Establishment

We examined the long-term success of prairie planting on a former strip mine in northeastern Illinois. The site was reclaimed and planted with prairie species in the 1970s. Total biomass increased over time, largely as a result of an increase in biomass of non-prairie species. Biomass of prairie species remained unchanged because of an increase in Panicum virgatum (switchgrass) offsetting decreases in Sorghastrum nutans (Indian grass). Total biomass was less than values published for other restored prairies (78 ± 4 g/m²to 298 ± 72 g/m² for our site, as opposed to 302-489 g/m² for the Trelease Prairie). Mycorrhizal inoculum potential (MIP) was variable across the site. There were also relatively few species of mycorrhizal fungi present as spores. Gigaspora sp., Scutellospora sp., Glomus sp., Glomus geosporum, and Glomus cf. fasciculatum were identified from spores. On a transect dominated by warm-season (C4) prairie grasses, MIP of rhizosphere soil collected under these species was lower than the MIP of rhizosphere soil collected under exotic cool-season (C3) grasses on a transect dominated by C3 species. On a transect with mixed warm-and cool-season vegetation, however, MIP did not differ under the two vegetation types. These results suggest that within-site patchiness rather than cover type is influencing MIP. Values of MIP were lower than those reported for native Illinois prairie.     

Patterns in intraspecific interaction strengths and the stability of food webs

A common approach to analyse stability of biological communities is to calculate the interaction strength matrix. Problematic in this approach is defining intraspecific interaction strengths, represented by diagonal elements in the matrix, due to a lack of empirical data for these strengths. Theoretical studies have shown that an overall increase in these strengths enhances stability. However, the way in which the pattern in intraspecific interaction strengths, i.e. the variation in these strengths between species, influences stability has received little attention. We constructed interaction strength matrices for 11 real soil food webs in which four patterns for intraspecific interaction strengths were chosen, based on the ecological literature. These patterns included strengths that were (1) similar for all species, (2) trophic level dependent, (3) biomass dependent, or (4) death rate dependent. These four patterns were analysed for their influence on (1) ranking food webs by their stability and (2) the response in stability to variation of single interspecific interaction strengths. The first analysis showed that ranking the 11 food webs by their stability was not strongly influenced by the choice of diagonal pattern. In contrast, the second analysis showed that the response of food web stability to variation in single interspecific interaction strengths was sensitive to the choice of diagonal pattern. Notably, stability could increase using one pattern and decrease using another. This result asks for deliberate approaches to choose diagonal element values in order to make predictions on how particular species, interactions, or other food web parameters affect food web stability.