Monitoring PDT by means of superficial reflectance spectroscopy.

Monitoring of relevant parameters during photodynamic therapy (PDT) and correlating these with treatment response is necessary to guarantee optimal and reproducible treatment outcome. In this paper we study the correlation between changes in the local tissue optical properties (absorption and scattering coefficients) during ALA-PDT and changes in PpIX fluorescence. The optical properties are measured extremely superficially by employing a single fiber for the delivery and collection of white light to and from the tissue. The measured reflectance spectrum is modeled in terms of four relevant parameters: blood saturation, relative blood volume fraction, scattering intensity and wavelength dependence of the scattering. All these parameters, except the relative blood volume fraction, are shown to correlate with the rate of photobleaching of PpIX, which in turn has previously been shown to correlate with the response of tissues to PDT. These results yield valuable insight in the behavior of these parameters during PDT and their suitability to predict PDT-response for other photosensitizers for which monitoring through photobleaching is not possible.

Assessment of bioconcentration and secondary poisoning of surfactants.

The relevance of the bioconcentration behaviour of surfactants for the secondary poisoning assessment and for the risk characterisation in the bird and mammalian food chain has been investigated. The approach used is described in the recently revised EU Technical Guidance Document for the Risk Assessment of Substances. The results demonstrate that, based on experimentally derived bioconcentration factors, environmental concentrations and effects in animals, there is a clear level of safety for both linear alkylbenzene sulphonate (LAS) and alcohol ethoxylates (AE), the most important surfactants by volume. To assess other surfactants used in detergents, a bioconcentration factor that would need to be attained for secondary poisoning to be of concern has been estimated from predicted environmental concentrations and known long-term effects data in animals. Based on the known structural similarity of these surfactants to LAS and AE and the ubiquitous nature of the enzymatic systems that are present in biotransformation processes in organisms, it is concluded that bioconcentration of these surfactants to these levels is highly unlikely. Therefore the potential for secondary poisoning effects of these surfactants is extremely low.

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 4. The influence of salt speciation, soil type, and sewage sludge on toxicity using the collembolan Folsomia fimetaria and the earthworm Aporrectodea caliginosa as test organisms.

Sewage sludge applied to agricultural soils often contains linear alkylbenzene sulfonates (LAS) in the range of 1 to 10 g/kg dry weight, and their toxicity to relevant soil organisms should, therefore, be assessed to ensure safe use of sewage sludge as a fertilizer. Studies of LAS toxicity to soil organisms are few, and to our knowledge, factors that may influence the toxicity in the field have not been studied in detail. In this paper, we report on the influence of speciation of LAS in the test solution added to soil (soluble Na-LAS vs poorly soluble Ca-LAS or Mg-LAS), the influence of soil type, and the modifying effects of sludge amendment on the toxicity of LAS. These issues were investigated using reproduction of Collembola and growth of juvenile earthworms as test parameters. Speciation of the LAS added to test soil did not have any influence on toxicity for any of the test species. Likewise, in three different agricultural soils (sand, loam, and clay), we found almost equal toxicities. The LAS added to test soil in a sludge-water suspension was equally toxic as when it was added in an aqueous solution. However, anaerobic incubation for 7 and 14 d of the LAS-sludge suspension (with no decay of LAS) caused the toxicity to increase almost threefold in both collembolan and earthworm. The relationships between soil constituents, bioavailability, and toxicity are also discussed.

X-ray crystal structures and conformational analysis of bicyclo[5.3.1]undec-1(11)-enes: twisting versus pyradidalization in anti-bredt olefins.

The synthesis of several 9,11,11-trihalo[5.3.1]propellanes and their 4-dimethylsila analogues is described. They solvolyze under formation of the corresponding isomeric 7,9,11- trihalobicyclo[5.3.l]undec-1(11)-enes which are "anti-Bredt" olefins with a strained trans double bond in a bridged eight-membered ring; in the presence of nucleophiles such as water or ethanol, the corresponding 7-hydroxy or 7-ethoxy derivatives, respectively, are obtained. On the basis of the X-ray crystal structures of four of these compounds (1a, 9a, 15, 17b), the effect of strain and of the substitution pattern on the degree of twisting and pyramidalization of the double bond is discussed.

Topical 5-aminolevulinic acid-photodynamic therapy of hairless mouse skin using two-fold illumination schemes: PpIX fluorescence kinetics, photobleaching and biological effect.

Light fractionation with dark periods of the order of hours has been shown to considerably increase the efficacy of 5-aminolevulinic acid-photodynamic therapy (ALA-PDT). Recent investigations have suggested that this increase may be due to the resynthesis of protoporphyrin IX (PpIX) during the dark period following the first illumination that is then utilized in the second light fraction. We have investigated the kinetics of PpIX fluorescence and PDT-induced damage during PDT in the normal skin of the SKH1 HR hairless mouse. A single illumination (514 nm), with light fluences of 5, 10 and 50 J cm-2 was performed 4 h after the application of 20% ALA, to determine the effect of PDT on the synthesis of PpIX. Results show that the kinetics of PpIX fluorescence after illumination are dependent on the fluence delivered; the resynthesis of PpIX is progressively inhibited following fluences above 10 J cm-2. In order to determine the influence of the PpIX fluorescence intensity at the time of the second illumination on the visual skin damage, 5 + 95 and 50 + 50 J cm-2 (when significantly less PpIX fluorescence is present before the second illumination), were delivered with a dark interval of 2 h between light fractions. Each scheme was compared to illumination with 100 J cm-2 in a single fraction delivered 4 or 6 h after the application of ALA. As we have shown previously greater skin damage results when an equal light fluence is delivered in two fractions. However, significantly more damage results when 5 J cm-2 is delivered in the first light fraction. Also, delivering 5 J cm-2 at 5 mW cm-2 + 95 J cm-2 at 50 mW cm-2 results in a reduction in visual skin damage from that obtained with 5 + 95 J cm-2 at 50 mW cm-2. A similar reduction in damage is observed if 5 + 45 J cm-2 are delivered at 50 mW cm-2. PpIX photoproducts are formed during illumination and subsequently photobleached. PpIX photoproducts do not dissipate in the 2 h dark interval between illuminations.

Terrestrial risk assessment for linear alkyl benzene sulfonate (LAS) in sludge-amended soils.

A comparison of the estimated environmental concentration and the effect concentrations (in the laboratory or field) in the receiving compartment form the basis of environmental risk assessments. This paper reviews processes that critically influence the fate of LAS in the terrestrial environment. Concentrations of LAS in sludge are quite high due to sorption to primary sludge, precipitation of Ca and Mg-salts of LAS, and lack of biodegradation under anaerobic digestion. This implies that when sludge is applied to soil e.g. as a fertilizer, considerable amounts of this important surfactant may enter the terrestrial environment. Influence of aerobic situations on LAS concentrations during sludge storage needs further research to allow incorporation into the risk assessment. Aerobic biodegradation in soil is considered the most important removal mechanism of LAS loading to the terrestrial environment through sludge-amendment. Sorption plays a role in determining the residence time of a chemical in the soil, hereby enabling more time for biodegradation to occur. In addition, sorption may affect the expression of effects of surfactants towards benthic and soil dwelling organisms and plants. Another factor that needs further attention is the form of LAS in the environment, which is not similar to the commercial material applied in detergents. The differential sorption and biodegradation of the LAS components lead to a shift in the alkyl chain length (homologue), and phenylisomer distribution towards increased hydrophobicity. Also, occurrence of Ca/Mg-salts in the environment versus the Na-salt for the commercial material critically impacts the extrapolation of effects data obtained in lab studies (mostly performed with the commercial material) to the field. The literature data were used in combination with strategies and methods provided by the European Union Technical Guidance Document in support of risk assessment of new and notified substances (1996) for the prediction of environmental concentrations of LAS entering the soil system through sludge applications. Soil biodegradation is an essential, necessary element for the PEC-calculations of LAS. The initial realistic worst case assessment presented indicates no human health risks exists with indirect exposure to LAS through either food or drinking water. Also, current LAS use does not pose a risk to terrestrial organisms such as plants and invertebrates.

A novel method to determine uptake and elimination kinetics of volatile chemicals in fish.

The development of an exposure system suitable for studying the uptake and elimination kinetics in fish of volatile chemicals is discussed. Static exposure of the fish is in a closed system containing water and air. Automated sampling and analysis of the air provides a concentration-time profile that is then fit to differential equations using numerical integration methods. Assumptions for the mathematical description of the system are a) instantaneous distribution of chemical between water and air and b) a first order one-compartment model describes the kinetics of chemical in fish. Uptake and elimination rate constants in fathead minnows (Pimephales promelas) were determined for a mixture of benzene, toluene, monochlorobenzene, monobromobenzene, and 1,3-dichlorobenzene. No significant biotransformation was observed for any of the compounds. Uptake rate constants increased with increasing octanol-water partition coefficient (Kow), whereas the elimination rate constants were inversely related to Kow.

Biotransformation of monochloroanilines in guppy, Poecilia reticulata.

1. Guppies (Poecilia reticulata) were exposed for 96 h to 4-chloroaniline (4CA), 3-chloroaniline (3CA) and 2-chloroaniline (2CA) in a static exposure system. 4-Chloroacetanilide (N4CA), 3-chloroacetanilide (N3CA), and 2-chloroacetanilide (N2CA) were identified in test water as the respective biotransformation products. 2. Dependence of biotransformation on aqueous concentration was investigated with 4CA. The measured biotransformation product in water (N4CA) increased with concentration increase of parent compound, 4CA, and with exposure time. 3. Quantitative differences in net acetylation of the three monochlorinated anilines were examined. The amount of acetanilide found in water increased in the following order: N2CA < N3CA < N4CA. Mass balance calculations indicated the highest loss of aniline occurred for 2CA. This compound, which is highest in toxicity, had the smallest amount of acetylated product formed. 4. This study proves, for the first time, that the acetylation reaction in fish is reversible. Quantitative differences in the net deacetylation of monochloroacetanilides were observed. The amount of deacetylated product found in water increased in the order 4CA < 3CA < 2CA.

N-acetyltransferase activity in rainbow trout liver and in vitro biotransformation of chlorinated anilines and benzenes in fish.

1. N-acetyl transferase activity in liver homogenate of rainbow trout (Oncorhynchus mykiss) was studied. Enzyme activity depends on the concentration of cofactor, has a broad pH and temperature optimum, is not linear with protein concentration within the whole range tested, and does not decrease upon storage at -70 degrees C. 2. In vitro biotransformation of several chlorinated anilines and benzenes was studied in liver homogenates of rainbow trout and swordtail (Xiphophorus helleri). Several phase I and II products were detected in the simple in vitro biotransformation assays using different cofactors NADPH-regenerating system and acetyl-CoA, respectively. Acetylation of di-ortho substituted anilines was not observed. 3. Apparent Vmax and Km values for the acetylation of trichloroanilines have been determined using rainbow trout liver homogenate. The rate or extent of N-acetylation is related to the structure and properties of the chlorinated anilines. 4. Comparison of the data for the two species showed that there are no apparent qualitative differences in the in vitro fate of the chlorinated anilines and benzenes studied. It is concluded that results obtained for these chemicals in the in vitro biotransformation assay can be extrapolated between the taxonomic families of Salmonidae and Poeciliidae. 5. The in vitro and in vivo N-acetylation of the chlorinated anilines turned out to be strikingly similar. Therefore, simple in vitro systems may be of use in assessing the potential of chemicals to bioconcentrate.

Toxicokinetics of aromatic amines in guppy, Poecilia reticulata.

In this study the elimination kinetics of several chlorinated anilines and a tetrachlorobenzene in guppy, Poecilia reticulata, have been determined. The elimination of all the chlorinated anilines in fish showed a bi-phasic pattern, whereas the elimination characteristics of the tetrachlorobenzene are in agreement with a one-compartment model. Elimination of the chlorinated anilines is faster than elimination of the tetrachlorobenzene. The small differences in octanol-water partition coefficient of these compounds can only in part describe the differences in elimination rate. The observed differences might be attributed to differing biotransformation rates.

Influence of survival time on the lethal body burden of 2,3,4,5-tetrachloroaniline in the guppy, Poecilia reticulata.

The lethal body burden (LBB) of 2,3,4,5-tetrachloroaniline in guppy (Poecilia reticulata), exposed for up to 14 days, has been determined at different aqueous concentrations of the test compound. It was found that the LBB is not constant during these 14 days, declining within the first 48 h to an approximately constant value. It is concluded that LBBs of organic compounds may not always be constant and may depend on survival time.

Bioconcentration of chlorinated anilines and benzene in fish: preliminary results.

1. Seven chlorinated anilines and one chlorinated benzene were tested for their ability to bioconcentrate in guppies (Poecilia reticulata) under different experimental conditions. 2. Interactions between compounds in a mixture influence the bioconcentration of some chlorinated anilines. These interactions result in either an increase or a decrease of bioconcentration, depending on the compound studied. 3. Exposure concentration can have an effect on the extent of bioconcentration of some chlorinated anilines.

Conservative management of ureteral tumor.

The traditional approach to transitional-cell carcinoma of the ureter has been nephroureterectomy with excision of a cuff of bladder. Evidence indicates that noninvasive transitional-cell tumor of the ureter is amenable to local resection. Five cases are presented in which transitional-cell carcinoma of the ureter was resected locally. None of the patients died from recurrence of their original tumor. It is suggested that the approach to ureteral tumor includes excisional biopsy with frozen section and reconstruction of ureteral continuity in the case of low-grade noninvasive transitional-cell tumor.