Nonequilibrium of organic compounds in sediment-water systems. Consequences for risk assessment and remediation measures.

In many cases, sediment risk assessment, and remediation rely on the assumption of equilibrium between chemical concentrations in sediment pore water and overlying surface water and thus rely on pore water concentrations only and do not additionally include assessment of the overlying water concentration. Traditionally, the validity of this assumption was insufficiently documented due to a lack of data. Recent studies using passive samplers, however, provided sufficient data for the first systematic evaluation of the extent of disequilibrium between sediment pore water and overlying surface water. Recent bioaccumulation studies reveal uncertainty as to which of these concentrations govern bioaccumulation by benthic organisms. Here, we provide the first review of studies measuring disequilibrium identifying general patterns and implications for the aforementioned areas of application. In most studies on water/sediment (dis)equilibrium, sediment pore water and overlying surface water are close to equilibrium. For lower molecular weight PAHs, overlying water concentrations tended to be relative low, which is tentatively ascribed to biodegradation in the water column. Substantial nonequilibrium was observed at some hot-spot locations such as in semistagnant harbors. In such cases, efficacy of sediment remediation measures to improve overlying water quality can be questioned because differences between overlying water concentrations at the hot-spots and those at reference locations typically are small. For nonequilibrium situations and some benthic taxa, exposure may be determined best by pore water concentrations. Improving our understanding in this area may further improve risk assessment of contaminated sediments.

Characterization of the inhibitory roles of RFRP3, the mammalian ortholog of GnIH, in the control of gonadotropin secretion in the rat: in vivo and in vitro studies.

RF-amide related peptides (RFRP), as putative mammalian orthologs of the avian gonadotropin-inhibitory hormone (GnIH), have been proposed as key regulators of gonadotropin secretion in higher vertebrates. Yet considerable debate has arisen recently on their physiological relevance and potential mechanisms and sites of action. Present studies were undertaken to further characterize the effects of RFRP on LH and FSH secretion by a combination of in vivo and in vitro approaches in male and female rats. Initial screening via intracerebroventricular (icv) administration of different analogs of RFRP1 (RFRP1-12 and RFRP1-20) and RFRP3 (RFRP3-8 and RFRP3-17), as well as the related neuropeptide FF (NPFF8), to gonadectomized (GNX) female rats evidenced significant, albeit modest, inhibitory effects on LH secretion only for RFRP3-8 and RFRP3-17, which were detectable at the high dose rage (1 nmol for RFRP3-8, 5 nmol for RFRP3-17). This moderate inhibitory action was also documented after icv administration of RFRP3-8 to intact and GNX male rats. In addition, systemic (intravenous) administration of RFRP3-8 decreased the circulating levels of both gonadotropins in GNX male rats. Likewise, RFRP3-8 inhibited basal and GnRH-stimulated LH secretion by pituitaries from GNX males in vitro. This inhibitory effect was blocked by the antagonist of RFRP receptors, RF9. In summary, our results support a putative inhibitory role of RFRP3 as ortholog of GnIH in the regulation of gonadotropin secretion in mammals, which appears to involve direct pituitary actions as well as potential central (hypothalamic) effects.

Characterization of the potent gonadotropin-releasing activity of RF9, a selective antagonist of RF-amide-related peptides and neuropeptide FF receptors: physiological and pharmacological implications.

Identification of RF-amide-related peptides (RFRP), as putative mammalian orthologs of the avian gonadotropin-inhibitory hormone, has drawn considerable interest on its potential effects and mechanisms of action in the control of gonadotropin secretion in higher vertebrates. Yet, these analyses have so far relied mostly on indirect approaches, while direct assessment of their physiological roles has been hampered by the lack of suitable antagonists. RF9 was recently reported as a selective and potent antagonist of the receptors for RFRP (RFRPR) and the related neuropeptides, neuropeptide FF (NPFF) and neuropeptide AF (NPFF receptor). We show here that RF9 possesses very strong gonadotropin-releasing activities in vivo. Central administration of RF9 evoked a dose-dependent increase of LH and FSH levels in adult male and female rats. Similarly, male and female mice responded to intracerebroventricular injection of RF9 with robust LH secretory bursts. In rats, administration of RF9 further augmented the gonadotropin-releasing effects of kisspeptin, and its stimulatory effects were detected despite the prevailing suppression of gonadotropin secretion by testosterone or estradiol. In fact, blockade of estrogen receptor-alpha partially attenuated gonadotropin responses to RF9. Finally, systemic administration of RF9 modestly stimulated LH secretion in vivo, although no direct effects in terms of gonadotropin secretion were detected at the pituitary in vitro. Altogether, these data are the first to disclose the potent gonadotropin-releasing activity of RF9, a selective antagonist of RFRP (and NPFF) receptors. Our findings support a putative role of the RFRP/gonadotropin-inhibitory hormone system in the central control of gonadotropin secretion in mammals and have interesting implications concerning the potential therapeutic indications and pharmacological effects of RF9.

Follicle-stimulating hormone responses to kisspeptin in the female rat at the preovulatory period: modulation by estrogen and progesterone receptors.

Ovulation is triggered by the preovulatory surge of gonadotropins that, in rodents, is defined by the concomitant rise in circulating LH and FSH at the afternoon of proestrus (primary surge), followed by persistently elevated FSH levels at early estrus (secondary surge). In recent years, kisspeptins, products of the KiSS-1 gene that act via G protein-coupled receptor 54, have emerged as an essential hypothalamic conduit for the generation of the preovulatory LH surge by conveying positive feedback effects of estradiol onto GnRH neurons, an event that involves not only estradiol-induced transcription of the KiSS-1 gene at the anteroventral periventricular nucleus but also its ability to modulate GnRH/LH responses to kisspeptin. However, little is known about the potential modulation of FSH responsiveness to kisspeptin by sex steroids in the cyclic female. We report herein analyses on the consequences of selective blockade of estrogen receptors (ER)-alpha and -beta, as well as progesterone receptor (PR), on the ovulatory surges of FSH and their modulation by kisspeptin. Antagonism of ERalpha or PR equally blunted the primary and secondary surges of FSH and nullified FSH responses to kisspeptin at the preovulatory period. Conversely, selective blockade of ERbeta failed to induce major changes in terms of endogenous FSH surges, yet it decreased FSH responses to exogenous kisspeptin. In contrast, FSH responses to GnRH were fully conserved after ERbeta blockade and partially preserved after inhibition of ERalpha and PR signaling. Finally, secondary FSH secretion was rescued by kisspeptin in females with selective blockade of ERalpha but not PR. In sum, our results substantiate a concurrent, indispensable role of ERalpha and PR in the generation of FSH surges and the stimulation of FSH responses to kisspeptin at the ovulatory period. In addition, our data suggest that ERbeta might operate as a subtle, positive modulator of the preovulatory FSH responses to kisspeptin, a role that is opposite to its putative inhibitory action on kisspeptin-induced LH secretion and might contribute to the dissociation of gonadotropin secretion at the ovulatory phase in the cyclic female rat.

Opposite roles of estrogen receptor (ER)-alpha and ERbeta in the modulation of luteinizing hormone responses to kisspeptin in the female rat: implications for the generation of the preovulatory surge.

Ovulation is triggered by the preovulatory rise of gonadotropins, which is in turn elicited by the preceding increase in circulating estrogen. Kisspeptins, ligands of G protein-coupled receptor 54 encoded by the KiSS-1 gene, have emerged as potent stimulators of GnRH/LH secretion, and KiSS-1 neurons at the anteroventral periventricular nucleus have been shown to be involved in the generation of preovulatory LH surge, estrogen being a potent elicitor of KiSS-1 gene expression selectively at the anteroventral periventricular nucleus. Whether, in addition to transcriptional effects, estrogen influences other aspects of kisspeptin-induced GnRH/LH release in the female remains unexplored. We provide herein evidence for the specific roles of estrogen receptor (ER)-alpha and ERbeta in the modulation of LH responses to kisspeptin and the generation of the preovulatory surge. Selective blockade of ERalpha in cyclic females blunted LH responses to kisspeptin, eliminated the endogenous preovulatory rise of LH, and blocked ovulation. In contrast, antagonism of ERbeta failed to cause major changes in terms of LH surge and ovulatory rate but significantly augmented acute LH responses to kisspeptin. Notably, defective LH secretion and ovulation after ERalpha blockade were not observed after GnRH stimulation, which elicited maximal acute (<2 h) LH responses regardless of ERalpha/ERbeta signaling. In addition, net LH secretion in response to kisspeptin was decreased by ovariectomy and increased after selective activation of ERalpha but not ERbeta. Altogether, our data document the prominent positive role of ERalpha in the regulation of GnRH/LH responsiveness to kisspeptin and, thereby, ovulation. In addition, our results disclose the putative function of ERbeta as negative modifier of GnRH/LH response to kisspeptin, a phenomenon that might contribute to partially restraining LH secretion at certain physiological states.

Comparison of key enzymes in the zebra mussel, Dreissena polymorpha, the earthworm Allolobophora chlorotica and Chironomus riparius larvae.

The levels of the enzymes, glutathione S-transferase, catalase, NAD(P)H-cytochrome c reductases, and DT-diaphorase were determined and compared in the tissues of three invertebrates commonly used in monitoring environmental quality: a freshwater mussel, Dreissena polymorpha, the earthworm Allolobophora chlorotica and the fourth instar of Chironomus riparius. It was found that the activities of GST, catalase, and NAD(P)-cytochrome c reductases were comparable in A. chlorotica and C. riparius, whereas comparatively a higher GST and a lower catalase activity was determined in the mussel tissues. DT-diaphorase was not detectable in A. chlorotica and the C. riparius larvae tissues, whereas this enzyme is present in the gills and the rest of soft mussel tissues (soft mussel tissues minus gills). It is suggested that the relatively low catalase activity observed in the tissues of the latter organism might be compensated by the presence of the antixidant role of DT-diaphorase. In addition, the inducibility of DT-diaphorase in D. polymorpha, by butylated hydroxyanisole (BHA) and lead (Pb) was investigated. Despite the bioaccumulation of both BHA (5.2+/-0.14 microgg(-1) wet weight) and Pb (233.7+/-0.95 mgkg(-1) dry weight) in the soft mussel tissues, the mussel DT-diaphorase was not induced. Although the activity of NADPH-cytochrome c (P-450) reductase was also not affected by these reagents, its activity was 2-fold higher in the gills than the rest of soft mussel tissues.

Differential responses of biomarkers in tissues of a freshwater mussel, Dreissena polymorpha, to the exposure of sediment extracts with different levels of contamination.

In this study, zebra mussels, D. polymorpha, were exposed to extracts of sediments obtained from two sites, a contaminated lake (Ketelmeer, Km) and a relatively clean lake (Drontenmeer, Dm). The main objective of this work was to investigate whether six selected biomarkers could discriminate between the two sediments. The selected biomarkers included phase I enzymes such as DT-diaphorase, NADPH-cytochrome c reductase, NADH-cytochrome c reductase, a phase II enzyme (glutathione S-transferase, GST), an antioxidant enzyme, catalase, and the total glutathione, reduced (GSH) and oxidized (GSSG). After a short (24 h) and a long-term (7 days) exposure, the levels of these biomarkers were measured in gills and the rest of soft mussel tissues (soft mussel tissue minus gills) and compared with control values. A decrease of GST level by 20% (P = 0.004) and a 4-fold decrease of total glutathione concentration relative to the control, were observed in the gills of mussels exposed to the more contaminated Km extract. No significant differences in the GST activities were observed in the gills of control and Dm extract-treated mussels (P = 0.23). Although the levels of catalase and NADH-cytochrome c reductase were, in the short-term exposure, unaffected, both activities were, in the long-term exposure, reduced in the gills of the mussels exposed to the contaminated Km extract, compared with control values, by 43% and 20%, respectively. The activities of DT-diaphorase and NADPH-cytochrome c reductase remained unaffected in all exposure conditions. However, the level of NADPH-cytochrome c reductase was found higher in gills than in the rest of soft mussel tissues. This difference in the ratio of the two reductases between the two tissues could account for the observed differential responses of the biomarkers.

In vivo exposure of Dreissena polymorpha mussels to the quinones menadione and lawsone: menadione is more toxic to mussels than lawsone.

The principal aim of this study was to assess whether the two quinones, menadione (2-methyl-1,4-naphthoquinone) and lawsone (2-hydroxy-1,4-naphthoquinone), elicit differential toxicity in mussels as has been reported for higher organisms. Therefore, the effects of short-term (48 h) and long-term (20 days) exposure of the two quinones at concentrations of 0.56 and 1 mg l(-1) to zebra mussels, Dreissena polymorpha, under laboratory conditions were studied. After the short-term exposure, the specific activities of the two-electron quinone oxidoreductase (DT-diaphorase) and the one-electron catalysing quinone reductases NADPH-cytochrome c reductase and NADH-cytochrome c reductase were determined in the gills and the rest of the soft tissues (soft mussel tissues minus the gills) of both treated and control mussels. At the higher concentrations of menadione and lawsone used, a significant reduction of the activity of NADPH-cytochrome c reductase in the gills and in the rest of the soft mussel tissues (by 33-34% and 31-43%, respectively) was observed. The activities of DT-diaphorase and NADH-cytochrome c reductase were not significantly affected. Interestingly, DT-diaphorase was observed in the gills, an organ requiring protection against antioxidants. Furthermore, a single-cell electrophoretic assay (comet assay) performed with gill cells to assess DNA damage by the quinones did not show any significant difference between the treated and the control organisms. This indicates that the formation of reactive species by the quinone metabolism in vivo in the mussels was possibly suppressed through the concerted action of DT-diaphorase and antioxidant enzymes. The results of in vitro experiments with gill extracts confirmed the protective role of DT-diaphorase. The rate of the two-electron quinone reduction was found to be five times that of the one-electron quinone reduction. The results of the long-term exposure unambiguously demonstrated that in mussels menadione, unlike in higher organisms, is more toxic than lawsone. The lack of detectability of xanthine oxidase in the mussel tissues could explain the comparatively lower toxicity of lawsone in the invertebtrate, lending support to a previous suggestion that xanthine oxidase might be responsible for the mechanism of toxicity of lawsone in higher organisms in vivo.

Menadione enhances oxyradical formation in earthworm extracts: vulnerability of earthworms to quinone toxicity.

NAD(P)H-cytochrome c reductase activities have been determined in the earthworms, L. rubellus and A. chlorotica, extracts. Menadione (0.35 mM, maximum concentration tested) was found to stimulate the rates of NADPH- and NADH-dependent cytochrome c reduction by three- and twofold, respectively. Superoxide dismutase (SOD) inhibited completely this menadione-mediated stimulation, suggesting that *O2- is involved in the redox cycling of menadione. However, SOD had no effect on the basal activity (activity in the absence of quinone) in the case of NADH-dependent cytochrome c reduction, whereas it partially inhibited the basal activity of NADPH-cytochrome c reduction. This indicates direct electron transfer in the former case and the formation of superoxide anion in the latter. DT-diaphorase, measured as the dicumarol-inhibitable part of menadione reductase activity, was not detectable in the earthworms' extracts. In contrast, it was found that DT-diaphorase represents about 70% of the menadione reductase activities in the freshwater mussel, Dreissena polymorpha. The results of this work suggest that earthworms, compared with mussels, could be more vulnerable to oxidative stress from quinones due to lack, or very low level of DT-diaphorase, an enzyme considered to play a significant role in the detoxification of quinones. On the contrary, mussels have efficient DT-diaphorase, which catalyzes two-electron reduction of menadione directly to hydroquinone, thus circumventing the formation of semiquinone.

Evidence for redox cycling of lawsone (2-hydroxy-1,4-naphthoquinone) in the presence of the hypoxanthine/xanthine oxidase system.

This study reports that lawsone (2-hydroxy-1,4-naphthoquinone) undergoes redox cycling in the presence of the hypoxanthine/xanthine oxidase system. The rate of cytochrome c reduction obtained in the presence of 80 microM lawsone was almost three times the rate of cytochrome c reduction measured in its absence. This increase in the rate of cytochrome c reduction was partially inhibited by superoxide dismutase, suggesting the involvement of O(2)(.-) in this process. It is remarkable to note that, even though lawsone is considered to be a non-redox-cycling quinone in vitro, this quinone was shown to be more toxic in vivo in rats than menadione, causing haemolytic anemia of an oxidative nature and renal damage. The view that this quinone is a non-redox-cycling quinone was based on the inability of one-electron-transferring flavoenzymes such as NADPH-cytochrome c reductase to reduce this naphthoquinone. Our finding that lawsone, like menadione, undergoes redox cycling in the presence of the hypoxanthine/xanthine oxidase system could explain the observed oxidative damage of tissues inflicted by this quinone in rats in vivo. Such an observation therefore reconciles the in vivo toxicity results of this naphthoquinone with those of in vitro experiments.

Resistant sorption of in situ chlorobenzenes and a polychlorinated biphenyl in river Rhine suspended matter.

The desorption kinetics of in situ chlorobenzenes (dichlorobenzenes, pentachlorobenzene and hexachlorobenzene) and 2,4,4'-trichlorobiphenyl (PCB-28) were measured with a gas-purge technique for river Rhine suspended matter sampled in Lobith, The Netherlands. This suspended matter is the main source of sediment accumulation in lake Ketelmeer. In lake Ketelmeer sediment earlier observations showed that slow and very slow fractions dominate the desorption profile. For the river Rhine suspended matter, only for PCB-28 a fast desorbing fraction of around 1.6% could be detected. The observed rate constants were on the average 0.2 h(-1) for fast desorption, 0.004 h(-1) for slow desorption, and 0.00022 h(-1) for very slow desorption. These values are in agreement with previous findings for the sediment from lake Ketelmeer and with available literature data on fast, slow, and very slow desorption kinetics. The results from this study show the similarity of desorption profiles between river Rhine suspended matter, and the top layer sediment from lake Ketelmeer. This indicates that slow and very slow fractions are already present in material forming the top layer of lake Ketelmeer, and were not formed after deposition of this material in the lake. The absence of detectable fast fractions for most compounds could be caused by the absence of recent pollution of the suspended matter. But, the observations may also be explained by a rapid disappearance of compounds from the fast fraction due to a combination of a high affinity of very slow sites for these compounds, and their relatively high volatility.

A simple Tenax extraction method to determine the availability of sediment-sorbed organic compounds.

A simple method to determine the availability of sediment-sorbed organic contaminants was developed and validated. For 10 polycyclic aromatic hydrocarbons, 4 polychlorinated biphenyls, and 9 chlorobenzenes in 6 sediments, we measured the fraction extracted by Tenax in 6 and 30 h. These fractions were compared with the rapidly desorbing fractions determined by consecutive Tenax extraction. Extraction by Tenax for 30 h completely removed the rapidly desorbing fraction plus some part of the slowly desorbing fraction. The fraction removed after 30 h was about 1.4 times the rapidly desorbing fraction. The fraction extracted by Tenax after 6 h is about 0.5 times the rapidly desorbing fraction for chlorobenzenes (CBs)/polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). The rapidly desorbing fraction probably represents the fraction of sorbed organic compound that poses actual risks for transport to (ground) water and determines the uptake by organisms and that can be microbially degraded. Extraction by Tenax for 6 h provides an easy way to address these issues more accurately than does the measurement of total concentrations.

Determination and theoretical aspects of the equilibrium between dissolved organic matter and hydrophobic organic micropollutants in water (Kdoc).

Literature on the equilibrium constant for distribution between dissolved organic carbon (DOC) (Kdoc) data of strongly hydrophobic organic contaminants were collected and critically analyzed. About 900 Kdoc entries for experimental values were retrieved and tabulated, including those factors that can influence them. In addition, quantitative structure-activity relationship (QSAR) prediction equations were retrieved and tabulated. Whether a partition or association process between the contaminant and DOC takes place could not be fully established, but indications toward an association process are strong in several cases. Equilibrium between a contaminant and DOC in solution was shown to be achieved within a minute. When the equilibrium shifts in time, this was caused by either a physical or chemical change of the DOC, affecting the lighter fractions most. Adsorption isotherms turned out to be linear in the contaminant concentration for the relevant DOC concentration up to 100 mg of C/L. Eighteen experimental methods have been developed for the determination of the pertinent distribution constant. Experimental Kdoc values revealed the expected high correlation with partition coefficients over n-octanol and water (Kow) for all experimental methods, except for the HPLC and apparent solubility (AS) method. Only fluorescence quenching (FQ) and solid-phase microextraction (SPME) methods could quantify fast equilibration. Only 21% of the experimental values had a 95% confidence interval, which was statistically significantly different from zero. Variation in Kdoc values was shown to be high, caused mainly by the large variation of DOC in water samples. Even DOC from one sample gave different equilibrium constants for different DOC fractions. Measured Kdoc values should, therefore, be regarded as average values. Kdoc was shown to increase on increasing molecular mass, indicating that the molecular mass distribution is a proper normalization function for the average Kdoc at the current state of knowledge. The weakly bound fraction could easily be desorbed when other adsorbing media, such as a SepPak column or living organism, are present. The amount that moves from the DOC to the other medium will depend, among other reasons, on the size of the labile DOC fraction and the equilibrium constant of the other medium. Variation of Kdoc with temperature turned out to be small, probably caused by a small enthalpy of transfer from water to DOC. Ionic strength turned out to be more important, leading to changes of a factor of 2-5. The direction of this effect depends on the type of ion. With respect to QSAR relationships between Kdoc and macroscopic or molecular descriptors, it was concluded that only a small number of equations are available in the literature, for apolar compounds only, and with poor statistics and predictive power. Therefore, a first requirement is the improvement of the availability and quality of experimental data. Along with this, theoretical (mechanistic) models for the relationship between DOC plus contaminant descriptors on the one side and Kdoc on the other should be further developed. Correlations between Kdoc and Kow and those between the soil-water partition constant (Koc) and Kow were significantly different only in the case of natural aquatic DOC, pointing at substantial differences between these two types of organic material and at a high correspondence for other types of commercial and natural DOC.

Field research for the authorisation of pesticides.

On request of the Dutch government a committee of the Health Council of the Netherlands has reviewed the role that results of field research in its broadest sense (i.e., including multi-species toxicity tests in the laboratory, research on model ecosystems et cetera) can play in ecotoxicological risk assessment for the authorisation of pesticides. The Committee believes that field research can provide valuable additional data about the exposure of non-target organisms and the resultant effects at population, community and ecosystem level. However, it frequently is unclear how these data might be used in reaching a decision about authorisation. To solve this problem, it is necessary to specify what is understood by "unacceptable damage". Both more clearly formulated protection goals of the government and a better understanding of the ecological significance of effects are needed to clarify this. Furthermore, the Committee points out that the statistical power of field trials must be sufficient to allow for the detection of changes that might be regarded as ecologically relevant. Finally, it recommends keeping a finger on the pulse in relation to authorised pesticides by monitoring their presence in environmental compartments and by investigating their role in suddenly occurring mortality among conspicuous animal species, such as birds, fish and honeybees. This kind of research forms a safety net for substances that have been wrongly authorised.

Slow desorption of PCBs and chlorobenzenes from soils and sediments: relations with sorbent and sorbate characteristics.

The kinetics of slow desorption were studied for four soils and four sediments with widely varying characteristics [organic carbon (OC) content 0.5-50%, organic matter (OM) aromatic content (7-37%)] for three chlorobenzenes and five polychlorinated biphenyls (PCBs). Slowly and very slowly desorbing fractions ranged from 1 to 50% (slow) and 3 to 40% (very slow) of the total amount sorbed, and were observed for all compounds and all soils and sediments. In spite of the wide variations in sorbate K(OW) (factor 1000) and sorbent characteristics, the rate constants of slow (k(slow), around 10(-3) h(-1)) and very slow (k(very slow), 10(-5)-10(-4) h(-1)) desorption appeared to be rather constant among the sorbates and sorbents (both within a factor of 5). There was a good correlation (r(2) above 0.9) between the distribution over the slow, very slow and rapid sediment fractions and log K(OC), indicating that sorbate hydrophobicity may be important for this distribution. No correlation could be found between sorbent characteristics [OC, N, and O in the organic matter, polarity index C/(N+O), OC aromaticity as determined by CP-MAS (13)C-NMR] and slow desorption parameters (slowly/very slowly desorbing fractions+corresponding rate constants). The absence of (1) a correlation between k(slow) and k(very slow), respectively, and OC content, and (2) the narrow range of k(slow) and k(very slow) values, indicates that intra-OM diffusion is not the mechanism of slow or very slow desorption, because on the basis of this mechanism it would be expected that increasing OC content would lead to longer diffusion pathlengths and, consequently, to smaller rate constants. In addition, it was tested whether differential scanning calorimetry would reveal a glass transition in the soils/sediments. In spite of the sensitivity of the equipment used (changes in heat flow in the micro-Watt range were measurable), a glass transition was not observed. This means that activation enthalpies of slow desorption can be calculated from desorption measurements at various temperatures. In the present study these values ranged from 60 to 100 kJ/mol among the various soils and sediments studied.

Redox effects on the excited-state lifetime in chlorosomes and bacteriochlorophyll c oligomers.

Oligomers of [E,E] BChl CF (8, 12-diethyl bacteriochlorophyll c esterified with farnesol (F)) and [Pr,E] BChl CF (analogously, M methyl, Pr propyl) in hexane and aqueous detergent or lipid micelles were studied by means of steady-state absorption, time-resolved fluorescence, and electron spin resonance spectroscopy. The maximum absorption wavelength, excited-state dynamics, and electron spin resonance (EPR) linewidths are similar to those of native and reconstituted chlorosomes of Chlorobium tepidum. The maximum absorption wavelength of oligomers of [E,E] BChl CF was consistently blue-shifted as compared to that of [Pr,E] BChl CF oligomers, which is ascribed to the formation of smaller oligomers with [E,E] BChl CF than [Pr,E] BChl CF. Time-resolved fluorescence measurements show an excited-state lifetime of 10 ps or less in nonreduced samples of native and reconstituted chlorosomes of Chlorobium tepidum. Under reduced conditions the excited-state lifetime increased to tens of picoseconds, and energy transfer to BChl a or long-wavelength absorbing BChl c was observed. Oligomers of [E,E] BChl CF and [Pr,E] BChl CF in aqueous detergent or lipid micelles show a similar short excited-state lifetime under nonreduced conditions and an increase up to several tens of picoseconds upon reduction. These results indicate rapid quenching of excitation energy in nonreduced samples of chlorosomes and aqueous BChl c oligomers. EPR spectroscopy shows that traces of oxidized BChl c radicals are present in nonreduced and absent in reduced samples of chlorosomes and BChl c oligomers. This suggests that the observed short excited-state lifetimes in nonreduced samples of chlorosomes and BChl c oligomers may be ascribed to excited-state quenching by BChl c radicals. The narrow EPR linewidth suggests that the BChl c are arranged in clusters of 16 and 6 molecules in chlorosomes of Chlorobium tepidum and Chloroflexus aurantiacus, respectively.

N-acetylcysteine pretreatment of cardiac surgery patients influences plasma neutrophil elastase and neutrophil influx in bronchoalveolar lavage fluid.

OBJECTIVE: Study of leukocyte activation and release of toxic mediators during extracorporeal circulation (ECC). ECC can be used to study the potential protective effect of a pharmacon against neutrophil-mediated lung injury. Clinical studies have indicated that N-acetylcysteine (NAC) may improve systemic oxygenation and reduce the need for ventilatory support when given to patients with acute lung injury. DESIGN: Cardiac surgery patients were pretreated with high-dose NAC in order to assess the potential role of NAC to interfere with neutrophil-mediated inflammation and lung injury. PATIENTS: 18 patients who underwent ECC: group 1 (n = 8) no premedication (only placebo); group 2 (n = 10) NAC (72 mg/kg i.v. as a bolus, later 72 mg/kg over 12 h). MEASUREMENTS AND RESULTS: In group 2, the partial pressure of oxygen in arterial blood/fractional inspired oxygen 4 h after surgery was significantly higher than in group 1 (213 +/- 31 vs 123 +/- 22; p = 0.044). NAC pretreatment prevented an increase in plasma neutrophil elastase activity (18.9 +/- 6.9 vs 49.9 +/- 5.6 ng/ml in group 1 at the end of ECC; p = 0.027). Release of myeloperoxidase (MPO) was not affected (group 1:1105 +/- 225 ng/ml vs group 2:1127 +/- 81 at the end of ECC; p = 0.63). At the end of ECC, total antigenic human neutrophil elastase (group 1:671 +/- 72 ng/ml vs group 2:579 +/- 134; p = 0.37) and complex formation between elastase and alpha 1-proteinase inhibitor were no different in the two groups. There were no significant difference in cellular composition and mediators in the lavage fluid, although values for total number of neutrophils, elastase, MPO and interleukin-8 were lower in group 2. CONCLUSION: Pretreatment with NAC may prevent lung injury by diminishing elastase activity. Since the release of mediators, especially MPO, is not affected, this diminished activity of elastase may be achieved by enhanced inactivation by antiproteases after initial treatment.

Self quenching of chlorosome chlorophylls in water and hexanol-saturated water.

The optical properties of a methyl ester homolog of bacteriochlorophylld (BChld M ) and bacteriochlorophyllc (BChlc) in H2O, hexanol-saturated H2O and methanol were studied by absorption, fluorescence emission, and circular dichroism (CD). In H2O, BChld M spontaneously forms an aggregate similar to that formed in hexane, with absorption maximum at 730 nm and fluorescence emission at 748 nm. For the pigment sample in hexanol-saturated H2O, while the absorption peaks at 661 nm, only slightly red-shifted compared to the monomer, the fluorescence emission is highly quenched. When diluted 2-3 fold with H2O, the absorption returns to around 720 nm, characteristic of an aggregate. The CD spectrum of the H2O aggregate exhibits a derivative-shaped feature with positive and negative peaks, while the amplitude is lower than that of chlorosomes. The Fourier transform infrared spectra of BChld M aggregates in H2O and hexane were measured. A 1644 cm(-1) band, indicative of a bonded 13(1)-keto group, is detected for both samples. A marker band for 5-coordinated Mg was observed at 1611 cm(-1) for the two samples as well. To study the kinetic behavior of the samples, both single-photon counting (SPC) fluorescence and transient absorption difference spectroscopic measurements were performed. For BChld M in hexanol-saturated H2O, a fast decay component with a lifetime of 10 to 14 ps was detected using the two different techniques. The fast decay could be explained by the concentration quenching phenomenon due to a high local pigment concentration. For the pigment sample in H2O, SPC gave a 16 ps component, whereas global analysis of transient absorption data generated two fast components: 3.5 and 26 ps. The difference may arise from the different excitation intensities. With a much higher excitation in the latter measurements, other quenching processes, e.g. annihilation, might be introduced, giving the 3.5 ps component. Finally, atomic force microscopy was used to examine the ultrastructure of BChld M in H2O and hexanol-saturated H2O. Pigment clusters with diameters ranging from 15 to 45 nm were observed in both samples.

Femtosecond probe of structural analogies between chlorosomes and bacteriochlorophyll c aggregates.

Bacteriochlorophyll c pigments extracted from light harvesting chlorosomes in green photosynthetic bacteria are known to self-assemble into aggregates whose electronic spectroscopy resembles that of intact chlorosomes. Femtosecond optical experiments reveal that the chlorosomes and their reconstituted aggregates exhibit closely analogous internal energy transfer kinetics and exciton state evolution. These comparisons furnish compelling new evidence that proteins do not exert a major local role in the BChl c antenna pigment organization of intact chlorosomes.