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Patterns of speciation in the Yucca moths: parallel species radiations within the Tegeticula yuccasella species complex.

The interaction between yuccas and yucca moths has been central to understanding the origin and loss of obligate mutualism and mutualism reversal. Previous systematic research using mtDNA sequence data and characters associated with genitalic morphology revealed that a widespread pollinator species in the genus Tegeticula was in fact a complex of pollinator species that differed in host use and the placement of eggs into yucca flowers. Within this mutualistic clade two nonpollinating "cheater" species evolved. Cheaters feed on yucca seeds but lack the tentacular mouthparts necessary for yucca pollination. Previous work suggested that the species complex formed via a rapid radiation within the last several million years. In this study, we use an expanded mtDNA sequence data set and AFLP markers to examine the phylogenetic relationships among this rapidly diverging clade of moths and compare these relationships to patterns in genitalic morphology. Topologies obtained from analyses of the mtDNA and AFLP data differed significantly. Both data sets, however, corroborated the hypothesis of a rapid species radiation and suggested that there were likely two independent species radiations. Morphological analyses based on oviposition habit produced species groupings more similar to the AFLP topology than the mtDNA topology and suggested the two radiations coincided with differences in oviposition habit. The evolution of cheating was reaffirmed to have evolved twice and the closest pollinating relative for one cheater species was identified by both mtDNA and AFLP markers. For the other cheater species, however, the closest pollinating relative remains ambiguous, and mtDNA, AFLP, and morphological data suggest this cheater species may be diverged based on host use. Much of the divergence in the species complex can be explained by geographic isolation associated with the evolution of two oviposition habits.

Animals↗

Anti-inflammatory and anti-arthritic effects of Yucca schidigera: a review.

Yucca schidigera is a medicinal plant native to Mexico. According to folk medicine, yucca extracts have anti-arthritic and anti-inflammatory effects. The plant contains several physiologically active phytochemicals. It is a rich source of steroidal saponins, and is used commercially as a saponin source. Saponins have diverse biological effects, including anti-protozoal activity. It has been postulated that saponins may have anti-arthritic properties by suppressing intestinal protozoa which may have a role in joint inflammation. Yucca is also a rich source of polyphenolics, including resveratrol and a number of other stilbenes (yuccaols A, B, C, D and E). These phenolics have anti-inflammatory activity. They are inhibitors of the nuclear transcription factor NFkappaB. NFkB stimulates synthesis of inducible nitric oxide synthase (iNOS), which causes formation of the inflammatory agent nitric oxide. Yucca phenolics are also anti-oxidants and free-radical scavengers, which may aid in suppressing reactive oxygen species that stimulate inflammatory responses. Based on these findings, further studies on the anti-arthritic effects of Yucca schidigera are warranted.

Journal Article↗

Patterns of genetic structure among populations of an oligophagous pollinating yucca moth (Tegeticula yuccasella).

Plant-insect associations have served as models for investigations of coevolution and the influence of biotic interactions on diversification. The pollination association between yuccas and yucca moths is a classic example of an obligate mutualism often suggested to have been affected by coevolution. Recent work has shown high host specificity in pollinating yucca moths, and here we use Tegeticula yuccasella, the species with the widest diet breadth, to ask how host specificity and isolation by distance contribute to specialization. Isolation by distance at a regional scale was observed in nucleotide variation within the mitochondrial gene cytochrome oxidase I (COI) (r =.294; P =.003). Host-related genetic structure (F(ct) = 0.08) was found to be slightly lower than the level of structure observed between eastern and western moth populations (F(ct) = 0.096). However, 56% of the COI haplotypes sampled from moths on Yucca filamentosa mapped to a host-specific clade in the haplotype network. Taken together, these results suggest that differentiation among T. yuccasella populations on alternative hosts is slight, but gene flow is influenced by both host association and geographic distance.

Animals↗

Tritium and 36Cl as constraints on fast fracture flow and percolation flux in the unsaturated zone at Yucca Mountain.

An analysis of tritium and 36Cl data collected at Yucca Mountain, Nevada suggests that fracture flow may occur at high velocities through the thick unsaturated zone. The mechanisms and extent of this "fast flow" in fractures at Yucca Mountain are investigated with data analysis, mixing models and several one-dimensional modeling scenarios. The model results and data analysis provide evidence substantiating the weeps model [Gauthier, J.H., Wilson, M.L., Lauffer, F.C., 1992. Proceedings of the Third Annual International High-level Radioactive Waste Management Conference, vol. 1, Las Vegas, NV. American Nuclear Society, La Grange Park, IL, pp. 891-989] and suggest that fast flow in fractures with minimal fracture-matrix interaction may comprise a substantial proportion of the total infiltration through Yucca Mountain. Mixing calculations suggest that bomb-pulse tritium measurements, in general, represent the tail end of travel times for thermonuclear-test-era (bomb-pulse) infiltration. The data analysis shows that bomb-pulse tritium and 36Cl measurements are correlated with discrete features such as horizontal fractures and areas where lateral flow may occur. The results presented here imply that fast flow in fractures may be ubiquitous at Yucca Mountain, occurring when transient infiltration (storms) generates flow in the connected fracture network.

Chlorine↗

A biosphere modeling methodology for dose assessments of the potential Yucca Mountain deep geological high level radioactive waste repository.

Recent developments in performance standards for proposed high level radioactive waste disposal at Yucca Mountain suggest that health risk or dose rate limits will likely be part of future standards. Approaches to the development of biosphere modeling and dose assessments for Yucca Mountain have been relatively lacking in previous performance assessments due to the absence of such a requirement. This paper describes a practical methodology used to develop a biosphere model appropriate for calculating doses from use of well water by hypothetical individuals due to discharges of contaminated groundwater into a deep well. The biosphere model methodology, developed in parallel with the BIOMOVS II international study, allows a transparent recording of the decisions at each step, from the specification of the biosphere assessment context through to model development and analysis of results. A list of features, events, and processes relevant to Yucca Mountain was recorded and an interaction matrix developed to help identify relationships between them. Special consideration was given to critical/potential exposure group issues and approaches. The conceptual model of the biosphere system was then developed, based on the interaction matrix, to show how radionuclides migrate and accumulate in the biosphere media and result in potential exposure pathways. A mathematical dose assessment model was specified using the flexible AMBER software application, which allows users to construct their own compartment models. The starting point for the biosphere calculations was a unit flux of each radionuclide from the groundwater in the geosphere into the drinking water in the well. For each of the 26 radionuclides considered, the most significant exposure pathways for hypothetical individuals were identified. For 14 of the radionuclides, the primary exposure pathways were identified as consumption of various crops and animal products following assumed agricultural use of the contaminated water derived from the deep well. Inhalation of dust (11 radionuclides) and external irradiation (1 radionuclide) were also identified as significant exposure modes. Contribution to the total flux to dose conversion factor from the drinking water pathway for each radionuclide was also assessed and for most radionuclides was found to be less than 10% of the total flux to dose conversion factor summed across all pathways. Some of the uncertainties related to the results were considered. The biosphere modeling results have been applied within an EPRI Total Systems Performance Assessment of Yucca Mountain. Conclusions and recommendations for future performance assessments are provided.

Computer Simulation↗

Administration of charcoal, Yucca schidigera, and zinc acetate to reduce malodorous flatulence in dogs.

OBJECTIVE: To determine whether feeding activated charcoal, Yucca schidigera, and zinc acetate would ameliorate the frequency and odor characteristics of flatulence in dogs. DESIGN: In vitro screening of active agents followed by a randomized controlled trial. ANIMALS: 8 adult dogs. PROCEDURE: A fecal fermentation system was used to assess the effects of activated charcoal, Yucca schidigera, and zinc acetate alone and in combination on total gas production and production of hydrogen sulfide, the primary determinant of flatus malodor in dogs. All 3 agents were subsequently incorporated into edible treats that were fed 30 minutes after the dogs ate their daily rations, and the number, frequency, and odor characteristics of flatulence were measured for 5 hours, using a device that sampled rectal gases and monitored hydrogen sulfide concentrations. RESULT: Total gas production and number and frequency of flatulence episodes were unaffected by any of the agents. Production of hydrogen sulfide in vitro was significantly reduced by charcoal, Yucca schidigera, and zinc acetate by 71, 38, and 58%, respectively, and was reduced by 86% by the combination of the 3 agents. Consumption of the 3 agents was associated with a significant decrease (86%) in the percentage of flatulence episodes with bad or unbearable odor and a proportional increase in the percentage of episodes of no or only slightly noticeable odor. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that activated charcoal, Yucca schidigera, and zinc acetate reduce malodor of flatus in dogs by altering the production or availability of hydrogen sulfide in the large intestine.

Animals↗

Effects of carbadox, copper, or Yucca shidigera extract on growth performance and visceral weight of young pigs.

Two identical trials, each with 128 crossbred weanling barrows (6.8 +/- .2 and 7.6 +/- .2 kg for Trials 1 and 2, respectively), were conducted. In each trial, pigs were allowed to consume ad libitum a 16% CP corn-soybean meal basal diet (B), B + 55 ppm of carbadox, B + 250 ppm of copper (Cu), or B + 125 ppm of Yucca shidigera extract for 56 d (four pens/diet; eight pigs/pen). At d 56 of the test, eight pigs/diet (two pigs/pen) were slaughtered for carcass and viscera measurements. Data of carbadox treatment in Trial 1 are excluded from this paper because of an error in mixing of the diet. In Trial 1, during the test period from d 0 to 28, pigs fed Cu had greater (P < .05) ADG and ADFI than those fed B, whereas pigs fed yucca extract had higher ADFI (P < .05) but similar ADG (P > .05) compared with those fed B. During the period from 29 to 56 d, ADG was similar among pigs fed different diets (P > .05) but ADFI was affected (P < .05) by diet (yucca extract > Cu > B). No differences (P > .05) among diets were detected for gain/feed (G/F) and visceral weights expressed as a percentage of slaughter BW. In Trial 2, during the first 28 d, the ADG, ADFI, and G/F responses of pigs to Cu, yucca extract, and B were similar to those observed in Trial 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Influence of yucca extract on ruminal metabolism in cows.

Two trials were conducted to determine the influence of yucca extract on ruminal digestion, fermentation, and ammonia patterns using ruminally and duodenally cannulated dairy cows. In Trial 1, urea at 0 or 1% of the diet and yucca extract at 0 or 4 g/d formed four dietary treatments in a 2 x 2 factorial arrangement. The experimental design was a 4 x 4 Latin square with 15-d periods. Duodenal digesta were sampled every 6 h during the last 4 d of each period to determine OM and ADF digestibilities and bacterial protein synthesis in the rumen using Cr2O3 and 15N markers. Ruminal digestibilities were (percentage): OM 46.3 vs 43.0%, and ADF 35.9 vs 41.4%, with or without Deodorase. Microbial protein entering the duodenum averaged 2.7 vs 3.1 kg/d for the respective treatments. Ruminal measurements were not affected by treatment (P > .10). In Trial 2, five cows were used in a 5 x 5 Latin square with 7-d periods. Treatments were 0, 2, 4, 6, and 8 g/d of yucca extract administered via ruminal cannulas. Ruminal fluid was sampled 0, 1, 2, 4, 7, 11, 16, and 22 h after feeding during the last 2 d of each period. Average ruminal NH3 N ranged from 31.4 to 35.4 mg/dL, pH 5.99 to 6.18, and total VFA from 120 to 129 mM, and all did not differ among treatments (P > .10). Yucca extract administered at 4 g/d did not significantly affect ruminal digestibilities of OM and ADF, and up to 8 g/d did not affect ruminal NH3, pH, or VFA.

Ammonia↗

Effects of Yucca shidigera extract and soluble protein on performance of cows and concentrations of urea nitrogen in plasma and milk.

Twelve multiparous Holstein cows averaging 122 d postpartum were used in a replicated Latin square design with 21-d periods. Cows were fed diets containing either low or high soluble protein supplemented with 0 or 9 g/d of Yucca shidigera extract per cow. Dry matter intake and yields of milk, fat, 3.5% fat-corrected milk, and total solids were not affected by treatment. High soluble protein increased percentages of fat and total solids in milk but decreased percentages of crude protein and true protein in milk. Yucca shidigera extract did not affect percentages or yields of milk components, ruminal NH3 N, or urea N in milk or plasma. High soluble protein tended to increase both ruminal pH and concentrations of ruminal NH3 N. Yucca shidigera extract did not affect ruminal pH or NH3 N. High soluble protein increased concentrations of urea N in plasma and milk when both were measured by enzymatic assay, but differences in milk urea N measured by mid infrared reflectance spectroscopy were not significant. Neither Yucca shidigera extract nor increased ruminally undegradable protein was beneficial in this trial when fed to cows milking 30 to 35 kg/d. Concentrations of urea N in milk as measured by mid infrared reflectance spectroscopy were higher than those measured by enzymatic assay, but measurements by mid infrared reflectance spectroscopy were not as sensitive to treatment differences as were measurements by enzymatic assay.

Ammonia↗

Spectrophotometric determination of saponin in Yucca extract used as food additive.

A spectrophotometric method was developed for the determination of saponin in Yucca extract or its preparation for food additive use. A saponin fraction of Yucca extract was prepared by column chromatography with porous polymer, and hydrolyzed with a 2 mol/L mixture of hydrochloric acid-ethanol (1 + 1) to generate sapogenin. Sapogenin amounts were determined by measuring absorbance at 430 nm, based on the color reactions with anisaldehyde, sulfuric acid, and ethyl acetate. Recoveries from Yucca extracts were 91.5-95.1%, and the detection limit was 10 microg/kg. Commercial Yucca extracts for food additive use were composed of 5.6-6.4% (w/w) saponin, making it a minor component.

Food Additives↗

Biogenesis and cytochemistry of unspecialized peroxisomes in root cortical cells of Yucca torreyi L.

Microbodies containing bipyramidal crystalline nucleoid inclusions occur within every cortical cell in roots of Yucca torreyi. Reaction product deposition attributable to catalase, glycolate oxidase, and urate oxidase activities are cytochemically localized to Yucca root microbodies and classifies them as unspecialized peroxisomes on the basis of their enzyme complement and tissue origin. Crystalline nucleoids do not stain for glycolate or urate oxidase activities, appearing as negatively-stained inclusions, but are apparently reactive for catalase activity. Development of unspecialized peroxisomes in Yucca roots is consistent with all evidence for glyoxysome and leaf-type peroxisome biogenesis from ER. Dilated ends of ER cisternae accumulate cytochemically detectable glycolate oxidase activity. After considerable dilation, paracrystalline precursors to nucleoids form within the bulge, and the inclusion enlarges to comprise the majority of peroxisomal volume. Peroxisomes that are not attached to ER are observed with high voltage electron microscopy and in serial thin sections, implying that eventually the budding peroxisomes are vesiculated. The functions of these unspecialized peroxisomes are suggested based upon cytochemical detection of their partial enzyme complement and their spatial and developmental timing relationships within developing Yucca root cortical parenchyma cells.

Alcohol Oxidoreductases↗

Inhibition of oxidative stress in blood platelets by different phenolics from Yucca schidigera Roezl. bark.

OBJECTIVE: We investigated the comparative effects of resveratrol (trans-3,4',5-trihydroxystilbene), trans-3,3',5,5'tetrahydroxy-4'-methoxystilbene, and yuccaols A and C isolated from the bark of Yucca schidigera on oxidative stress in resting blood platelets and blood platelets activated by different agonists (thrombin or thrombin receptor activating peptide). METHODS AND RESULTS: Tested phenolics (1-25 microgram/mL) reduced, to different degrees, the level of reactive oxygen species measured by the luminol-dependent chemiluminescence and changed the production of O(2)(-) measured by the reduction of cytochrome c in resting blood platelets. They also inhibited the generation of free radicals in blood platelets activated by thrombin (P < 0.05) or thrombin receptor activating peptide (P < 0.05). Treatment of platelets with resveratrol or yuccaols A and C at the concentration of 25 microgram/mL increased (statistically non-significant) the level of thiobarbituric acid reactive substances in these cells (P > 0.05), whereas trans-3,3',5,5'tetrahydroxy-4'-methoxystilbene and the alcohol yucca extract reduced lipid peroxidation in blood platelets (P < 0.05). CONCLUSIONS: Resveratrol and other phenolic compounds from the bark of Yucca schidigera inhibiting free radical generation in blood platelets may be beneficial in protecting against cardiovascular diseases when hyperactivity of platelets is observed.

Animals↗

Limiting cheaters in mutualism: evidence from hybridization between mutualist and cheater yucca moths.

Mutualisms are balanced antagonistic interactions where both species gain a net benefit. Because mutualisms generate resources, they can be exploited by individuals that reap the benefits of the interaction without paying any cost. The presence of such 'cheaters' may have important consequences, yet we are only beginning to understand how cheaters evolve from mutualists and how their evolution may be curtailed within mutualistic lineages. The yucca-yucca moth pollination mutualism is an excellent model in this context as there have been two origins of cheating from within the yucca moth lineage. We used nuclear and mitochondrial DNA markers to examine genetic structure in a moth population where a cheater species is parapatric with a resident pollinator. The results revealed extensive hybridization between pollinators and cheaters. Hybrids were genetically intermediate to parental populations, even though all individuals in this population had a pollinator phenotype. The results suggest that mutualisms can be stable in the face of introgression of cheater genes and that the ability of cheaters to invade a given mutualism may be more limited than previously appreciated.

Adaptation, Biological↗

Preliminary 3-D site-scale studies of radioactive colloid transport in the unsaturated zone at Yucca Mountain, Nevada.

The U.S. Department of Energy (DOE) is actively investigating the technical feasibility of permanent disposal of high-level nuclear waste in a repository to be situated in the unsaturated zone (UZ) at Yucca Mountain (YM), Nevada. In this study we investigate, by means of numerical simulation, the transport of radioactive colloids under ambient conditions from the potential repository horizon to the water table. The site hydrology and the effects of the spatial distribution of hydraulic and transport properties in the Yucca Mountain subsurface are considered. The study of migration and retardation of colloids accounts for the complex processes in the unsaturated zone of Yucca Mountain, and includes advection, diffusion, hydrodynamic dispersion, kinetic colloid filtration, colloid straining, and radioactive decay. The results of the study indicate that the most important factors affecting colloid transport are the subsurface geology and site hydrology, i.e., the presence of faults (they dominate and control transport), fractures (the main migration pathways), and the relative distribution of zeolitic and vitric tuffs. The transport of colloids is strongly influenced by their size (as it affects diffusion into the matrix, straining at hydrogeologic unit interfaces, and transport velocity) and by the parameters of the kinetic-filtration model used for the simulations. Arrival times at the water table decrease with an increasing colloid size because of smaller diffusion, increased straining, and higher transport velocities. The importance of diffusion as a retardation mechanism increases with a decreasing colloid size, but appears to be minimal in large colloids.

Colloids↗

The saturated zone at Yucca Mountain: an overview of the characterization and assessment of the saturated zone as a barrier to potential radionuclide migration.

The US Department of Energy is pursuing Yucca Mountain, Nevada, for the development of a geologic repository for the disposal of spent nuclear fuel and high-level radioactive waste, if the repository is able to meet applicable radiation protection standards established by the US Nuclear Regulatory Commission and the US Environmental Protection Agency (EPA). Effective performance of such a repository would rely on a number of natural and engineered barriers to isolate radioactive waste from the accessible environment. Groundwater beneath Yucca Mountain is the primary medium through which most radionuclides might move away from the potential repository. The saturated zone (SZ) system is expected to act as a natural barrier to this possible movement of radionuclides both by delaying their transport and by reducing their concentration before they reach the accessible environment. Information obtained from Yucca Mountain Site Characterization Project activities is used to estimate groundwater flow rates through the site-scale SZ flow and transport model area and to constrain general conceptual models of groundwater flow in the site-scale area. The site-scale conceptual model is a synthesis of what is known about flow and transport processes at the scale required for total system performance assessment of the site. This knowledge builds on and is consistent with knowledge that has accumulated at the regional scale but is more detailed because more data are available at the site-scale level. The mathematical basis of the site-scale model and the associated numerical approaches are designed to assist in quantifying the uncertainty in the permeability of rocks in the geologic framework model and to represent accurately the flow and transport processes included in the site-scale conceptual model. Confidence in the results of the mathematical model was obtained by comparing calculated to observed hydraulic heads, estimated to measured permeabilities, and lateral flow rates calculated by the site-scale model to those calculated by the regional-scale flow model. In addition, it was confirmed that the flow paths leaving the region of the potential repository are consistent with those inferred from gradients of measured head and those independently inferred from water-chemistry data. The general approach of the site-scale SZ flow and transport model analysis is to calculate unit breakthrough curves for radionuclides at the interface between the SZ and the biosphere using the three-dimensional site-scale SZ flow and transport model. Uncertainties are explicitly incorporated into the site-scale SZ flow and transport abstractions through key parameters and conceptual models.

Forecasting↗

The site-scale saturated zone flow model for Yucca Mountain: calibration of different conceptual models and their impact on flow paths.

This paper presents several different conceptual models of the Large Hydraulic Gradient (LHG) region north of Yucca Mountain and describes the impact of those models on groundwater flow near the potential high-level repository site. The results are based on a numerical model of site-scale saturated zone beneath Yucca Mountain. This model is used for performance assessment predictions of radionuclide transport and to guide future data collection and modeling activities. The numerical model is calibrated by matching available water level measurements using parameter estimation techniques, along with more informal comparisons of the model to hydrologic and geochemical information. The model software (hydrologic simulation code FEHM and parameter estimation software PEST) and model setup allows for efficient calibration of multiple conceptual models. Until now, the Large Hydraulic Gradient has been simulated using a low-permeability, east-west oriented feature, even though direct evidence for this feature is lacking. In addition to this model, we investigate and calibrate three additional conceptual models of the Large Hydraulic Gradient, all of which are based on a presumed zone of hydrothermal chemical alteration north of Yucca Mountain. After examining the heads and permeabilities obtained from the calibrated models, we present particle pathways from the potential repository that record differences in the predicted groundwater flow regime. The results show that Large Hydraulic Gradient can be represented with the alternate conceptual models that include the hydrothermally altered zone. The predicted pathways are mildly sensitive to the choice of the conceptual model and more sensitive to the quality of calibration in the vicinity on the repository. These differences are most likely due to different degrees of fit of model to data, and do not represent important differences in hydrologic conditions for the different conceptual models.

Calibration↗

Colloid transport and deposition in water-saturated Yucca Mountain tuff as determined by ionic strength.

Colloid mobility and deposition were determined in model systems consisting of quartz sand or crushed Yucca Mountain tuff, latex microspheres (colloidal particles), and simulated groundwater. Ionic strength (I) was manipulated as a first step in defining limiting conditions for colloid transport in a system modeled after geochemical conditions at the Yucca Mountain site. Solutions of deionized water (DI), 0.1x, 1x, and 10x (the ionic strength of simulated groundwater) (I = 0.0116 M) were used in saturated columns under steady-state flow conditions. Separate experiments with conservative tracers indicated stable hydrodynamic conditions that were independent of I. Colloids were completely mobile (no deposition) in the DI and 0.1x solutions; deposition increased to 11-13% for 1x and to 89-97% for 10x treatments with similar results for sand and tuff. Deposition was described as a pseudo-first-order process; however, a decreasing rate of deposition was apparent for colloid transport at the 10x condition through the tuff. A linear dependence of colloid removal (extent and deposition rate coefficient) on I is illustrated for the model Yucca Mountain system and for a glass-KCl system reported in the literature. This simple relationship for saturated systems may be useful for predicting deposition efficiencies under conditions of varying ionic strength.

Colloids↗

Plutonium oxidation and subsequent reduction by Mn(IV) minerals in Yucca Mountain tuff.

Plutonium oxidation state distribution on Yucca Mountain tuff and synthetic pyrolusite (beta-MnO2) suspensions was measured using synchrotron X-ray micro-spectroscopy and microimaging techniques as well as ultrafiltration/solventextraction techniques. Plutonium sorbed to the tuff was preferentially associated with manganese oxides. For both Yucca Mountain tuff and synthetic pyrolusite, Pu(IV) or Pu(V) was initially oxidized to more mobile Pu(V/VI), but over time, the less mobile Pu(IV) became the predominant oxidation state of the sorbed Pu. The observed stability of Pu(IV) on oxidizing surfaces (e.g., pyrolusite), is proposed to be due to the formation of a stable hydrolyzed Pu(IV) surface species. These findings have important implications in estimating the risk associated with the geological burial of radiological waste in areas containing Mn-bearing minerals, such as at the Yucca Mountain or the Hanford Sites, because plutonium will be predominantly in a much less mobile oxidation state (i.e., Pu(IV)) than previously suggested (i.e., Pu(V/VI).

Hydrogen-Ion Concentration↗