Preparation of Cross-Linked Monodisperse Poly(acrylic acid) Particles by Precipitation Polymerization.

Water-soluble polymers such as poly(acrylic acid) (PAA) are widely applied in dispersants, thickeners, flocculants, and superabsorbent polymers. Here, submicrometer-sized monodisperse PAA particles were successfully prepared by precipitation polymerization in acetonitrile at 55 °C with 2,2'-azobis(2,4-dimethylvaleronitrile) as an initiator. In a medium with a high dielectric constant, the particles were stabilized by electrostatic repulsion without a colloidal stabilizer during the polymerization. Moreover, the obtained PAA particles were spontaneously cross-linked in the absence of a cross-linker. The degree of the cross-linking was strongly dependent on the amount of water (500-10,000 ppm) in the polymerization medium. The PAA particles swelled more with acrylic acid with an increase in the amount of water. Thus, in the case of high water content, the particles would become the main polymerization loci, which in turn results in the cross-linking structures owing to the α-hydrogen abstraction of PAA chains during the polymerization.

Major and trace elements in the surface water of Tonle Sap Lake, Mekong River, and other tributary rivers in Cambodia.

To evaluate the seasonal water circulation of Tonle Sap Lake and its tributary rivers in Cambodia, the spatial distribution patterns of major and trace elements in surface water were investigated. Based on the similarity of the dissolved elemental concentrations, the water samples were mainly divided into the three groups: samples with relatively high percentages of Ca, Mo, and Sb (Subcluster B1); samples with high Si, Al, and Fe (B2); and samples with high Na, K, and Mg (B3). During the rainy season, the elemental composition of lake water (B1) appeared to be greatly influenced by the intrusion of water from the Mekong River (B1) through the Tonle Sap River (B1). During the dry season, the type of lake water shifted to B3, suggesting that the lake water stored during the rainy season was replaced by inflow from other tributaries and groundwater in its vicinity. Thus, the seasonal changes in the elemental composition of the lake water were largely controlled by surface water and groundwater circulation. The dissolved As concentration was higher in the lake water and during the dry season than that in the river water and during the rainy season, indicating the discharge of As from the lake's bottom sediment during the dry season. Although the redox cycling of Fe and Mn appeared to be less important due to the shallow water depth in the lake, there are potential risks of As poisoning induced by the formation of an anoxic water mass and increment in the concentration of phosphorus if eutrophication continues to progress.

Preparation of Poly(Ionic Liquid) Particles with Anionic Side Chain by Dispersion Polymerization.

Micrometer-sized poly(ionic liquid) (PIL) particles with an anionic side chain, poly(1-butyl-3-methylimidazolium 3-sulfopropyl methacrylate) (poly([Bmim][SPMT])), are successfully prepared by dispersion polymerization at 60 °C in ethanol/ethyl acetate (2/8, w/w) with poly(vinylpyrrolidone) as a stabilizer. However, the obtained particles do not maintain the particulate state during drying at room temperature due to poly([Bmim][SPMT])'s low glass-transition temperature, below room temperature. In order to prevent coalescence and maintain particle shape, a cross-linking monomer is added after the nucleation stage of dispersion polymerization. Moreover, Fourier-transform infrared spectroscopy (FT-IR) demonstrates cation exchange between the obtained particles and metal cations (Ag+ ).

Geographic variation of Sr and S isotope ratios in bottled waters in Japan and sources of Sr and S.

The geographic distribution of the stable isotope ratio of Sr (87Sr/86Sr) is useful for identifying the provenance of drinking water, organisms, and agricultural products. To explore how environmental factors influence this ratio, we determined the 87Sr/86Sr, sulfur isotope ratio (δ34S), and elemental composition of 208 commercially available bottled waters in Japan, and compared them with the 87Sr/86Sr values of groundwater, rainwater, rocks, and vegetables from similar localities. We classified the bottled waters into seven categories based on the geology of their source aquifers. The 87Sr/86Sr and concentrations of several elements (Ca, K, Si, Rb, and Ba) of the bottled waters differed with the aquifer rock type and were well correlated with the 87Sr/86Sr of reported spring waters, indicating that bottled water, like other groundwater, reflects the lithology and chemistry of its aquifer. The 87Sr/86Sr of bottled water, taken as a proxy for groundwater, showed regional variations consistent with those of rocks and vegetables, demonstrating that an 87Sr/86Sr map of water and agricultural products has value for determining the production areas of these substances. However, the value and range of 87Sr/86Sr differed among these three materials, and the degree of the difference between groundwater and rock depended on the rock type. An analysis of geochemical data from rocks and rainwater suggests that groundwater contains Sr and additional S derived from atmospheric sources such as rainwater and sea salt aerosols. The atmospheric contribution of Sr to groundwater is greatest in areas of siliceous sedimentary rocks with low Sr concentrations and high resistance to weathering. A weak correlation of δ34S of bottled waters with 87Sr/86Sr indicates the usefulness of maps of combined Sr and S isotope ratios for groundwater provenance. This study shows that for accurate provenance determinations of groundwater, organisms, and agricultural products, it is essential to better quantify the contributions of Sr and S from the atmosphere as well as from rocks and human activities.

Variation of strontium stable isotope ratios and origins of strontium in Japanese vegetables and comparison with Chinese vegetables.

To evaluate the utility of 87Sr/86Sr ratio for determining the geographical provenance of vegetables, we compared 87Sr/86Sr ratios and Sr concentrations in five vegetable species grown in Japan and China, and we also examined the relationships between 87Sr/86Sr ratios in vegetables, the soil-exchangeable pool, irrigation water, and fertilizer from 20 Japanese agricultural areas. The vegetable 87Sr/86Sr ratios in Japan were similar for all species within a given agricultural area, but tended to be low in northeast Japan and high in southwest Japan. The median 87Sr/86Sr ratio in Japanese vegetables was similar to that in fertilizer, suggesting that in addition to rock-derived Sr, vegetables contain Sr derived from fertilizers. In most cases, the 87Sr/86Sr ratios for the Japanese and Chinese vegetables differed by approximately 0.710. Linear discriminant analysis using both 87Sr/86Sr and the Sr concentration allowed more accurate discrimination between vegetables from the two countries.

Tracing the Geographical Origin of Onions by Strontium Isotope Ratio and Strontium Content.

The strontium (Sr) isotope ratio ((87)Sr/(86)Sr) and Sr content were used to trace the geographical origin of onions from Japan and other countries, including China, the United States of America, New Zealand, Australia, and Thailand. The mean (87)Sr/(86)Sr ratio and Sr content (dry weight basis) for onions from Japan were 0.70751 and 4.6 mg kg(-1), respectively, and the values for onions from the other countries were 0.71199 and 12.4 mg kg(-1), respectively. Linear discriminant analysis was performed to classify onions produced in Japan from those produced in the other countries based on the Sr data. The discriminant equation derived from linear discriminant analysis was evaluated by 10-fold cross validation. As a result, the origins of 92% of onions were correctly classified between Japan and the other countries.

Potential uses of stable isotope ratios of Sr, Nd, and Pb in geological materials for environmental studies.

The ratios of stable isotopes of certain elements in rocks and minerals have strong regional characteristics that are reflected in atmospheric components, in water, and in the living organisms that form Earth's surface environment as well as in agricultural and fishery products. Geologically derived stable isotope ratios can be used as a tracer for the source of many kinds of substances, with current geochemical techniques allowing the precise determination of numerous stable isotope ratios in both natural and manmade objects. This review presents examples of the use of stable isotopes as tracers within diverse dynamic ecosystems, focusing on Sr isotopes but also including examples of Nd and Pb isotopic analysis, and reviewing the potential of this technique for a wide range of environmental research, including determining the geographic origin of food and archeological materials.

Carbon isotope ratios of human tooth enamel record the evidence of terrestrial resource consumption during the Jomon period, Japan.

OBJECTIVE: Archaeological remains strongly suggest that the Holocene Japanese hunter-gatherers, the Jomon people, utilized terrestrial plants as their primary food source. However, carbon and nitrogen isotope analysis of bone collagen indicates that they primarily exploited marine resources. We hypothesize that this inconsistency stems from the route of protein synthesis and the different proportions of protein-derived carbon in tooth enamel versus bone collagen. Carbon isotope ratios from bone collagen reflect that of dietary protein and may provide a biased signal of diet, whereas isotope ratios from tooth enamel reflect the integrated diet from all macronutrients (carbohydrates, lipids, and proteins). METHODS: In order to evaluate the differences in inferred diet between the archaeological evidence and bone collagen isotope data, this study investigated carbon isotopes in Jomon tooth enamel from four coastal sites of the Middle to Late-Final Jomon period (5,000-2,300 years BP). RESULTS: Carbon isotope ratios of human teeth are as depleted as coeval terrestrial mammals, suggesting that C3 plants and terrestrial mammals were major dietary resources for the Jomon people. Dietary dependence on marine resources calculated from enamel was significantly lower than that calculated from bone collagen. The discrepancy in isotopic ratios between enamel and collagen and the nitrogen isotope ratio in collagen shows a negative correlation on individual and population levels, suggesting diets with variable proportions of terrestrial and marine resources. CONCLUSION: This study highlights the usefulness of coupling tooth enamel and bone collagen in carbon isotopic studies to reconstruct prehistoric human diet. Am J Phys Anthropol 158:300-311, 2015. © 2015 Wiley Periodicals, Inc.

Oral Astaxanthin Supplementation Prevents Peritoneal Fibrosis in Rats.

BACKGROUND: Preventing peritoneal damage during peritoneal dialysis is critical. Reactive oxygen species (ROS) have an important role in peritoneal damage; however, few studies have investigated this. We aimed to determine the effects of oral astaxanthin (AST) supplementation in a peritoneal fibrosis (PF) rat model. METHODS: Thirty-seven Sprague-Dawley rats were divided into 5 groups: Control 1 (fed a normal diet without stimulation), Control 2 (fed an AST-supplemented diet without stimulation), Group 1 (fed a normal diet with 8% chlorhexidine gluconate [CG] stimulation for 3 weeks), Group 2 (fed a 0.06% AST-supplemented diet with CG stimulation), and Group 3 (fed a 0.06% AST-supplemented diet that was initiated 4 weeks before CG stimulation). Peritoneal fibrosis, vascular proliferation, and fibrosis-related factor expression were examined. RESULTS: Peritoneal thickness was significantly suppressed by AST supplementation. Astaxanthin diminished the number of CD68-, 8-hydroxy-2'-deoxyguanosine (8-OHdG)-, and monocyte chemoattractant protein-1 (MCP-1)-positive cells. Type 3 collagen, tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and MCP-1 mRNA expression was significantly lower in Group 3 than in Group 1. Increased transforming growth factor-ß (TGF-ß) and Snail mRNA expression, vascular density, and the number of α-smooth muscle actin (α-SMA)-positive cells were also decreased in Group 3. CONCLUSION: Astaxanthin suppressed PF development through the inhibition of inflammation and oxidation in PF rats. It appears that the anti-oxidative agent AST may be useful for the prevention of peritoneal damage.

Adipose-derived mesenchymal stem cells transplantation facilitate experimental peritoneal fibrosis repair by suppressing epithelial-mesenchymal transition.

BACKGROUND: Prevention or reversal of peritoneal damage is critical in peritoneal dialysis. Although autologous cell transplantation has beneficial effects on tissue repair in various organs, few studies have investigated the effects of transplantation of adipose-derived mesenchymal stem cells (ASCs) on peritoneal fibrosis (PF). Thus, we examined the mechanism of facilitated peritoneal reconstruction induced by ASC transplantation on chlorhexidine gluconate (CG)-induced PF in rats. METHODS: To induce PF in rats, continuous-infusion pumps containing 8 % CG were placed in the abdominal cavity for 21 days. The pumps were removed on day 22 and ASCs were immediately injected into the peritoneal cavity. Morphological alterations and mRNA expression levels of fibrosis-related factors were examined on days 29 and 35. RESULTS: ASC transplantation significantly facilitated peritoneal repair. mRNA expression of tumor necrosis factor-α, interleukin-1ß, monocyte chemotactic protein-1, and epithelial-mesenchymal transition (EMT) markers such as Snail and α-smooth muscle actin were suppressed, whereas that of vascular endothelial growth factor (VEGF) and platelet-derived growth factor-BB (PDGF-BB) were overexpressed after ASC transplantation. Immunofluorescence indicated that some transplanted ASCs expressed VEGF and PDGF-BB and differentiated into vascular cells. CONCLUSIONS: ASC transplantation facilitates peritoneal repair by suppressing EMT and modulating inflammation and angiogenesis during the early phase of tissue repair in experimental PF.

Comparison between the fixation of peritoneal dialysis catheters to the peritoneal wall and the conventional placement technique: clinical experience and follow-up of a new implant technique for peritoneal dialysis catheters.

Peritoneal dialysis (PD) catheters often become severely dislocated, which may lead to malfunction. With the aim of preventing this complication, we have developed a simple method of fixing the catheter downwards in the peritoneal cavity (fixation technique), a technique that does not require a laparoscope. Sixteen patients were implanted using the conventional placement technique and 25 patients were implanted using the fixation technique. The location of the catheter tip was classified from grade 1 (downward, normal) to 5 (dislocated). The frequency of dislocation (defined as the extended time and/or decrease in volume when draining the PD solution) was measured for both the fixation technique and conventional placement technique. There was a significant difference in grade between the fixation technique (2.72 ± 1.01) and conventional technique (3.92 ± 1.31). The time until first dislocation was significantly different between the fixation technique (59.3 ± 48.1 days) and conventional technique (8.8 ± 14.6 days). The time until any dislocation was significantly different between the fixation technique (69.2 ± 41.9 days) and conventional technique (12.9 ± 13.7 days). Complications were not significantly different between the fixation technique and conventional technique. The fixation technique appears to be simple, safe, and useful for preventing severe dislocation and for lengthening the time until dislocation in PD patients.

Paracrine effects of transplanted mesothelial cells isolated from temperature-sensitive SV40 large T-antigen gene transgenic rats during peritoneal repair.

BACKGROUND: The prevention and restoration of peritoneal damage is a critical mission in peritoneal dialysis (PD). Transplantation of mesothelial cells has been suggested to suppress peritoneal injury during PD. Few studies have examined the efficacy and safety of cell transplantation. We evaluated the paracrine effects of mesothelial transplantation during peritoneal repair using immortalized temperature-sensitive mesothelial cells (TSMCs) in chlorhexidine gluconate (CG)-induced peritoneal fibrosis rats. METHODS: Continuous-infusion pumps containing 8% CG were placed into the abdominal cavity for 21 days. After the removal of the pumps, the TSMCs were injected into the peritoneal cavity at Day 22 (Tx-1 group) or 29 (Tx-2 group). Morphological findings and mRNA expressions of regeneration-related factors were examined at Days 22, 29 and 35. RESULTS: Peritoneal thickness was aggravated in the Tx-1 group. Levels of transforming growth factor (TGF)-ß, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 mRNA in the Tx-1 group at Day 35 were comparable with those at Day 22. The levels of Snail, B-Raf and ERK-1, markers of epithelial to mesenchymal transition and of the RAS/MAPK pathway in the Tx-1 group, were significantly higher than those in the Tx-2 group. TGF-ß and VEGF were produced from the transplanted mesothelial cells and the surrounding cells in the Tx-1 group. CONCLUSION: It appears that the paracrine effect of transplanted mesothelial cells during peritoneal repair is associated with its surrounding condition. It is important to determine the most appropriate time for developing peritoneal repair through mesothelial transplantation.

Pentraxin 3 as a new biomarker of peritoneal injury in peritoneal dialysis patients.

It is well known that bioincompatible peritoneal dialysate plays a central role in the development of peritoneal fibrosis. Peritoneal inflammation continues even after the cessation of peritoneal dialysate stimulation. It is important to establish the definition of persistent inflammation in the peritoneal cavity at the cessation of peritoneal dialysis (PD). The objective of the present study was to determine whether pentraxin 3 (PTX3) in peritoneal effluent (PE) may be a new biomarker in PD patients. Serum, PE, and peritoneal specimens were obtained from 50 patients with end-stage kidney disease at Juntendo University Hospital. Samples of 19 patients were obtained at the initiation of PD and those of 31 patients at the cessation of PD. PTX3, high-sensitivity CRP, and MMP-2 and IL-6 were analyzed. An immunohistological examination using an anti-PTX3 antibody was performed. Expressions of PTX3 were observed in endothelial cells, fibroblasts, and mesothelial cells in the peritoneum. The PTX3 level in PE at the cessation of PD was significantly higher than that at the initiation of PD. Effluent PTX3 levels in patients with a history of peritonitis or a PD duration of more than 8 years were significantly higher than those in patients without peritonitis or patients with a PD duration of <8 years. The PTX3 level was significantly correlated with MMP-2 and IL-6 levels in PE, as well as the thickness of the submesothelial compact zone and the vasculopathy. It appears that PTX3 may be a new biomarker of peritoneal inflammation and progressive fibrosis.

Time series analysis for the estimation of tidal fluctuation effect on different aquifers in a small coastal area of Saijo plain, Ehime prefecture, Japan.

Considering the current poor understanding of the seawater-freshwater (SW-FW) interaction pattern at dynamic hydro-geological boundary of coastal aquifers, this work strives to study tidal effect on groundwater quality using chemical tracers combined with environmental isotopes. In situ measurement data of electrical conductivity and groundwater level along with laboratory measurement data of hydro-chemical species were compared with tidal level data measured by Hydrographic and Oceanographic Department, Saijo City, Japan for time series analysis. Result shows that diurnal tides have significant effect on groundwater level as well as its chemical characteristics; however, the magnitude of effect is different in case of different aquifers. Various scatter diagrams were plotted in order to infer mechanisms responsible for water quality change with tidal phase, and results show that cations exchange, selective movement and local SW-FW mixing were likely to be the main processes responsible for water quality changes. It was also found that geological structure of the aquifers is the most important factor affecting the intensity of tidal effect on water quality.

Evaluation of the Impacts of Marine Salts and Asian Dust on the Forested Yakushima Island Ecosystem, a World Natural Heritage Site in Japan.

To elucidate the influence of airborne materials on the ecosystem of Japan's Yakushima Island, we determined the elemental compositions and Sr and Nd isotope ratios in streamwater, soils, vegetation, and rocks. Streamwater had high Na and Cl contents, low Ca and HCO(3) contents, and Na/Cl and Mg/Cl ratios close to those of seawater, but it had low pH (5.4 to 7.1), a higher Ca/Cl ratio than seawater, and distinct (87)Sr/(86)Sr ratios that depended on the bedrock type. The proportions of rain-derived cations in streamwater, estimated by assuming that Cl was derived from sea salt aerosols, averaged 81 % for Na, 83 % for Mg, 36 % for K, 32 % for Ca, and 33 % for Sr. The Sr value was comparable to the 28 % estimated by comparing Sr isotope ratios between rain and granite bedrock. The soils are depleted in Ca, Na, P, and Sr compared with the parent materials. At Yotsuse in the northwestern side, plants and the soil pool have (87)Sr/(86)Sr ratios similar to that of rainwater with a high sea salt component. In contrast, the Sr and Nd isotope ratios of soil minerals in the A and B horizons approach those of silicate minerals in northern China's loess soils. The soil Ca and P depletion results largely from chemical weathering of plagioclase and of small amounts of apatite and calcite in granitic rocks. This suggests that Yakushima's ecosystem is affected by large amounts of acidic precipitation with a high sea salt component, which leaches Ca and its proxy (Sr) from bedrock into streams, and by Asian dust-derived apatite, which is an important source of P in base cation-depleted soils.

Evolution model of δ³4S and δ¹8O in dissolved sulfate in volcanic fan aquifers from recharge to coastal zone and through the Jakarta urban area, Indonesia.

The sources of sulfate in an aquifer system, and its formation/degradation via biogeochemical reactions, were investigated by determining sulfate isotope ratios (δ³4S(SO4) and δ¹8O(SO4) in dissolved sulfate in groundwater from the Jakarta Basin. The groundwater flow paths, water ages, and geochemical features are well known from previous studies, providing a framework for the groundwater chemical and isotopic data, which is supplemented with data for spring water, river water, hot spring water, seawater, detergents, and fertilizers within the basin. The sulfate isotope composition of groundwater samples varied widely from -2.9‰ to +33.4‰ for δ³4S(SO4) and +4.9‰ to +17.8‰ for δ¹8O(SO4) and changed systematically along its flow direction from the mountains north to the coastal area. The groundwater samples were classified into three groups showing (1) relatively low and narrow δ(34)S(SO4) (+2.3‰ to +7.6‰) with low and varied δ¹8O(SO4) (+4.9‰ to +12.9‰) compositions, (2) high and varied δ³4S(SO4) (+10.2‰ to +33.4‰) with high δ¹8O(SO4) (+12.4‰ to +17.3‰) compositions, and (3) low δ³4S(SO4) (< +6.1‰) with high δ¹8O(SO4) (up to +17.8‰) compositions. These three types of groundwater were observed in the terrestrial unconfined aquifer, the coastal unconfined and confined aquifers, and the terrestrial confined aquifer, respectively. A combination of field measurements, concentrations, and previously determined δ¹5N(NO3) data, showed that the observed isotopic heterogeneity was mainly the result of contributions of pollutants from domestic sewage in the rural area, mixing of seawater sulfate that had experienced previous bacterial sulfate reduction in the coastal area, and isotopic fractionation during the formation of sulfate through bacterial disproportionation of elemental sulfur. Our results clearly support the hypothesis that human impacts are important factors in understanding the sulfur cycle in present-day subsurface environments. A general model of sulfate isotopic evolution along with groundwater flow has rarely been proposed, due to the complicated hydrogeological research setting that causes varied isotope ratios, although its understanding has recently received great attention. This pioneer study on a simple volcanic fan aquifer system with a well-understood groundwater flow mechanism provides a useful model for future studies.

Human impacts on groundwater flow and contamination deduced by multiple isotopes in Seoul City, South Korea.

The influence of human activities on the flow system and contamination of groundwater were investigated in Seoul City, South Korea, one of the largest Asian cities, using a combination of isotopes (deltaD, T, delta15N, delta18O, delta34S, and 87Sr/86Sr). Eighteen representative groundwater and river water samples, which were collected over a wide area of the city, were compared with previously reported data. The distribution of stable isotopes (deltaD and delta18O) with groundwater potential data shows that recharged groundwater from either the surrounding mountainous area as well as the Han River and other surface streams discharged towards the northern-central part of the city, where a subway tunnel pumping station is located. It is suggested from T values (3.3 to 5.8 T.U.) that groundwater was recharged in the last 30 to 40 years. The delta34S and delta15N of SO4(2-) and NO3- data were efficiently used as indicators of contamination by human activities. These isotopes clarified that the contribution of anthropogenic contaminants i.e., industrial and household effluents, waste landfills, and fertilizers, are responsible for the enrichment by SO4(2-) (>30 ppm as SO4(2-)) and NO3- (>20 ppm as NO3-) of groundwater. The 87Sr/86Sr values of groundwater vary (0.71326 to 0.75058) in accordance with the host rocks of different origins. Mineral elements such as Ca are also suggested to be derived naturally from rocks. The groundwater under Seoul City is greatly affected by transportation of pollutants along the groundwater flow controlled by subway tunnel pumping, contributing to the degradation of water quality in urbanized areas.

Use of stable nitrogen isotope signatures of riparian macrophytes as an indicator of anthropogenic N inputs to river ecosystems.

Deterioration of aquatic ecosystems resulting from enhanced anthropogenic N loading has become an issue of increasing concern worldwide, and methods are needed to trace sources of N in rivers. Because nitrate from sewage is enriched in 15N relative to nitrate from natural soils, delta(15)N values of stream nitrate (delta(15)Nnitrate) should be an appropriate index of anthropogenic N loading to rivers, as should the delta(15)N values of riparian plants (delta(15)Nplant) because they are consumers of nitrate. We determined the delta(15)N values of stream nitrate and six species of riparian macrophytes in 31 rivers in the Lake Biwa Basin in Japan. We then tested the correlation between these values and various land-use parameters, including the percentage of land used for residential and agricultural purposes as well as for natural areas. These delta(15)N values were significantly positively correlated with land use (%) that had a high N load (i.e., residential or agricultural use) and significantly negatively correlated with forest (%). These findings indicate that delta(15)N values of stream nitrate and riparian plants might be good indicators of anthropogenic inputs of nitrogen.

Reverse evolution of armor plates in the threespine stickleback.

Faced with sudden environmental changes, animals must either adapt to novel environments or go extinct. Thus, study of the mechanisms underlying rapid adaptation is crucial not only for the understanding of natural evolutionary processes but also for the understanding of human-induced evolutionary change, which is an increasingly important problem [1-8]. In the present study, we demonstrate that the frequency of completely plated threespine stickleback fish (Gasterosteus aculeatus) has increased in an urban freshwater lake (Lake Washington, Seattle, Washington) within the last 40 years. This is a dramatic example of "reverse evolution,"[9] because the general evolutionary trajectory is toward armor-plate reduction in freshwater sticklebacks [10]. On the basis of our genetic studies and simulations, we propose that the most likely cause of reverse evolution is increased selection for the completely plated morph, which we suggest could result from higher levels of trout predation after a sudden increase in water transparency during the early 1970s. Rapid evolution was facilitated by the existence of standing allelic variation in Ectodysplasin (Eda), the gene that underlies the major plate-morph locus [11]. The Lake Washington stickleback thus provides a novel example of reverse evolution, which is probably caused by a change in allele frequency at the major plate locus in response to a changing predation regime.