Structures and redox characteristics of electron-deficient vanadyl phthalocyanines.

The first single-crystal X-ray structures of substituted vanadyl phthalocyanine materials reveal the high-valence vanadium ions (denoted as V(IV)), whose coordination by a highly electron-deficient ligand is facilitated by an axial oxo group. The metal center of the hydrophilic V═O core, encapsulated in F-rich hydrophobic pockets, reaches a coordination number of 6 by binding an additional H(2)O that, in turn, hydrogen-bonds with ketones, resulting in solvent-induced variable solid-state architectures. Fluoroalkyl (R(f)) ligand substituents hinder π-π stacking interactions and favor ordered long-range packing, as well as the facile formation of film materials that exhibit high thermal stability and oxidation resistance. Reversible redox chemistry and spectroscopic studies in both solution and the solid-state indicate single-site isolation in both phases and an R(f)-induced propensity for electron uptake and inhibition of electron loss. Repeated redox cycles reorganize the thin films to accommodate Li(+) ions and facilitate their migration. The facile reduction, combined with high stability and ease of sublimation imparted by the R(f) scaffold that suppresses oxidations, recommends the new materials for sensors, color displays, electronic materials, and redox catalysts, as well as other applications.

Broadening the reactivity spectrum of a phthalocyanine catalyst while suppressing its nucleophilic, electrophilic and radical degradation pathways.

A robust molecule that resists degradation via nucleophilic, electrophilic and radical attacks is described. Coordinated O(2) is reduced catalytically, producing efficiently thyil radicals in spite of the extreme electronic deficiency of the catalyst.

Effects of water chemistry on the chronic toxicity of lead to the cladoceran, Ceriodaphnia dubia.

As the first step toward parameterization of a chronic lead (Pb) biotic ligand model (BLM) for Ceriodaphnia dubia, 7-d toxicity tests were performed in waters modified to evaluate the influences of hardness, DOM (as Suwannee River NOM and Aldrich humic acid (HA)), pH (buffered with 4 mM MOPS) and alkalinity on the chronic toxicity of Pb. Calculated EC(20)s for the control base water test and each of the most extreme modified test waters were as follows in µg L(-1) Pb (95% confidence interval): base water control=45 (14-53), 5 mM CaSO(4)=22 (12-30), 32 mg L(-1) DOM=523 (388-573), 2.5 mM NaHCO(3)=73 (21-120) and pH 6.4 buffered with MOPS=3.9 µg L(-1) Pb (1-5). Results indicate that hardness does not protect against chronic toxicity of Pb to C. dubia, whereas HA does protect at the highest concentration tested (597 µM). Additionally, our findings suggest that low pH increases the chronic toxicity of Pb whereas increased alkalinity is protective. The findings reported herein support the need for a chronic Pb BLM as an alternative approach to hardness-based regulations.

Thiocyanate, calcium and sulfate as causes of toxicity to Ceriodaphnia dubia in a hard rock mining effluent.

A series of Toxicity Identification Evaluations (TIEs) to identify the cause(s) of observed toxicity to Ceriodaphnia dubia have been conducted on a hard rock mining effluent. Characteristic of hard rock mining discharges, the effluent has elevated (∼3000 mg l(-1)) total dissolved solids (TDS) composed primarily of Ca(2+) and SO(4)(2-). The effluent typically exhibits 6-12 toxic units (TUs) when tested with C. dubia. Phase I and II toxicity identification evaluations (TIEs) indicated Ca(2+) and SO(4)(2-) contributed only ∼4 TUs of toxicity, but this was likely an underestimate due to problems with simulating the supersaturated CaSO(4) concentrations in the effluent. Treatment of the effluent with BaCO(3) to precipitate Ca(2+) and SO(4)(2-) revealed that these ions contribute ∼6 TUs of the observed toxicity, but the remaining source(s) of toxicity (up to 6 TUs) remained unidentified. Subsequent investigations identified thiocyanate (SCN(-)) in the effluent at 100-150 µM. Toxicity tests reveal that C. dubia are sensitive to SCN(-) with an estimated IC25 of 8.3 µΜ for reproduction in moderately hard water suggesting between 12 and 18 TUs of toxicity in the effluent. Additional experiments demonstrated that SCN(-) toxicity is reduced in the high TDS matrix of the mining effluent. Testing of a mock effluent simulating the major ion and SCN(-) concentrations resulted in 10.4 TUs, suggesting that Ca(2+), SO(4)(2-) and SCN(-) are the three toxicants present in this effluent. This research suggests SCN(-) may be a more common cause of toxicity in mining effluents than is generally recognized.

The effects of total dissolved solids on egg fertilization and water hardening in two salmonids--Arctic Grayling (Thymallus arcticus) and Dolly Varden (Salvelinus malma).

Previous studies have indicated that salmonid fertilization success may be very sensitive to elevated concentrations of total dissolved solids (TDS) with effects at concentrations as low as 250mgl(-1) being reported. However, interpretation of these studies is complicated by poor control performance and variable concentration response relationships. To address this, a series of experiments were performed to evaluate TDS effects on Arctic Grayling (Thymallus arcticus) and Dolly Varden (Salvelinus malma) fertilization success and identify possible mechanisms for previously observed test variability and any observed effects of TDS. Results indicate that some of the experiments reported here were likely confounded by extended milt holding times prior to experiment initiation. Milt holding times >6h were shown to significantly reduce control fertilization and corresponding concentration response relationships were variable. When milt holding time was minimized during fertilization experiments, consistent control performance with >90% control fertilization was achieved and consistent concentration response relationships were observed for both species examined. Experiments performed under these conditions indicate that Arctic Grayling and Dolly Varden fertilization success is not sensitive to elevated TDS with EC20s (concentration causing 20% effect) of >2782 and >1817mgl(-1) (the highest concentrations tested), respectively. However, TDS was shown to significantly affect embryo water absorption during the water hardening phase immediately following fertilization. The lowest observable effect concentrations (LOECs) for this endpoint were 1402 and 964mgl(-1) for Arctic Grayling and Dolly Varden, respectively. The effect of reduced embryo turgidity, due to impaired water absorption, on resistance to mechanical damage under real world conditions needs further investigation in order to understand the implications of this observed effect.

Rational design of a reactive yet stable organic-based photocatalyst.

Zinc perfluoro-fluoroalkyl-phthalocyanine, synthesized in high yield, does not exhibit electron loss, does not aggregate in solution, is photostable and produces (1)O(2) in very high quantum yields. Aerobic photo-oxygenation of an external substrate occurs without catalyst self-oxidation. The encapsulation of a metal center in a refractory organic environment could guide the design of other viable catalysts for oxygenation of substrates either for synthesis or for oxidative destruction of organic or biological molecules, under reaction conditions that include the use of only air and light.

Life history and aging of captive-reared California sea hares (Aplysia californica).

Although the California sea hare, Aplysia californica, is well known from neurobiological studies and is raised in the laboratory for this purpose, various aspects of its life history in the laboratory, such as aging dynamics, are unknown. Therefore we collected life history data on 4 cohorts of eggs from hatchery-reared animals and performed an actuarial analysis of mortality data. Temperature was controlled at 13 to 15 degrees C, the photoperiod was a 14:10-h light:dark cycle, and the seawater O2 concentration, pH, and salinity were held at optimized levels. The feeding protocol for 3 cohorts was unrestricted access to the red macroalga Gracilaria ferox, whereas the remaining cohort was fed standard hatchery rations of G. ferox 4 times per week. Growth was sigmoidal in each cohort and resulted in linear growth rates of 1.25 to 3.62 g/d during the exponential phase; these rates were not influenced by feeding level. Sexual maturity occurred at approximately 160 g, at ages ranging from 144 to 241 d. Egg production was highly variable in the different cohorts. Mean lifespan of cohorts fed ad libitum was approximately 228 d. In contrast, the cohort fed standard rations lived an average of 375 d and showed a lower initial mortality rate, suggesting that calorie restriction on a single-species diet prolongs lifespan in California sea hares.

Chronic toxicity of lead to three freshwater invertebrates--Brachionus calyciflorus, Chironomus tentans, and Lymnaea stagnalis.

Chronic lead (Pb) toxicity tests with Brachionus calyciflorus, Chironomus tentans, and Lymnaea stagnalis were performed in artificial freshwaters. The no-observable-effect concentration (NOEC), lowest-observable-effect concentration (LOEC), and calculated 20% effect concentration (EC20) for the rotifer B. calyciflorus were 194, 284, and 125 microg dissolved Pb/L, respectively. The midge C. tentans was less sensitive, with NOEC and LOEC of 109 and 497 microg dissolved Pb/L, respectively, and the snail L. stagnalis exhibited extreme sensitivity, evident by NOEC, LOEC, and EC20 of 12, 16, and < 4 microg dissolved Pb/L, respectively. Our findings are presented in the context of other reports on chronic Pb toxicity in freshwater organisms. The L. stagnalis results are in agreement with a previous report on pulmonate snails and should be viewed in the context of current U.S. Environmental Protection Agency (U.S. EPA) hardness adjusted water quality criteria of 8 microg Pb/L. The present findings and earlier reports indicate that freshwater pulmonate snails may not be protected by current regulatory standards. Measurements of whole-snail Na+ and Ca2+ concentrations following chronic Pb exposure revealed that Na+ homeostasis is disturbed by Pb exposure in juvenile snails in a complicated pattern, suggesting two physiological modes of action depending on the Pb exposure concentration. Substantially reduced growth in the snails that exhibit very high Ca2+ requirements may be related to reduced Ca2+ uptake and thereby reduced shell formation.

Temperature effects on growth, maturation, and lifespan of the california sea hare (Aplysia californica).

We conducted a hatchery growth study to describe the variability in growth rates, spawning, and mortality of Aplysia californica in regard to rearing temperature. Animals were housed at a standard hatchery density of five animals per cage, at temperatures of 13, 15, 18, and 21 degrees Celsius. Animals reared at 13 or 15 degrees C grew as much as four times as large, lived twice as long, matured later, and spawned longer than did animals reared at 18 or 21 degrees C. At age 170 to 205 days the fastest growth rates occurred at 18 and 21 degrees C, and the slowest at 13 degrees C. As animals at 18 and 21 degrees C reached sexual maturity at ages 190 to 197 days, or approximately 60% through their lifespans, their growth rates slowed such that by age 260 days, the fastest growth rate was at 13 degrees C, and the slowest was at 21 degrees C. Animals reared at 13 and 15 degrees C reached sexual maturity at 242 and 208 days, respectively, or at approximately 40% of their life spans. Lifespan and maximum average animal weight were significantly inversely correlated with temperature (P

Identification of S-glutathionylated cellular proteins during oxidative stress and constitutive metabolism by affinity purification and proteomic analysis.

Redox modification of proteins is proposed to play a central role in regulating cellular function. However, high-throughput techniques for the analysis of the redox status of individual proteins in complex mixtures are lacking. The aim was thus to develop a suitable technique to rapidly identify proteins undergoing oxidation of critical thiols by S-glutathionylation. The method is based on the specific reduction of mixed disulfides by glutaredoxin, their reaction with N-ethylmaleimide-biotin, affinity purification of tagged proteins, and identification by proteomic analysis. The method unequivocally identified 43 mostly novel cellular protein substrates for S-glutathionylation. These include protein chaperones, cytoskeletal proteins, cell cycle regulators, and enzymes of intermediate metabolism. Comparisons of the patterns of S-glutathionylated proteins extracted from cells undergoing diamide-induced oxidative stress and during constitutive metabolism reveal both common protein substrates and substrates failing to undergo enhanced S-glutathionylation during oxidative stress. The ability to chemically tag, select, and identify S-glutathionylated proteins, particularly during constitutive metabolism, will greatly enhance efforts to establish posttranslational redox modification of cellular proteins as an important biochemical control mechanism in coordinating cellular function.

S-glutathionylation of glyceraldehyde-3-phosphate dehydrogenase: role of thiol oxidation and catalysis by glutaredoxin.

The findings in this article illustrate the complexity residing in the regulation of reversible S-glutathionylation of proteins, such as GAPDH. This is clearly reflected in the design of suitable experimental approaches designed to cope with the interaction of several redox-dependent factors. Clear interactions are demonstrated between oxidative modification of GAPDH and its subsequent S-glutathionylation. Similarly, a redox interaction between GSSG and GAPDH with Grx as the catalyst is shown, suggesting that the Grx molecule may participate in catalytic S-glutathionylation in intact cells. Furthermore, Grx itself can readily undergo S-glutathionylation, indicating the potential for regulation of this catalyst of the reversible S-glutathionylation of other proteins. The methodologies detailed in this work may provide a good reference point for other attempts to elucidate the mechanism of reversible S-glutathionylation of purified proteins in a manner that more closely resembles the situation arising in intact cells during the generation of oxidative stress.