Leaching potential of some phenylureas and their main metabolites through laboratory studies

BACKGROUND, AIMS AND SCOPE: Laboratory studies were conducted with the aim of defining the leaching potential of some phenylureas and their metabolites. A first study was performed for calculating their leaching index (as GUS) on the base of intrinsic properties: persistence (as DT50) and mobility (as Koc) in soil. Another study consisted of aged column leaching experiments whose meaning was to semi-quantify the occurrence of the tested compounds in the leachates, so simulating in field conditions. METHODS: The tested compounds were: diuron, linuron and monolinuron (parents); 3,4-dichloroaniline (DCA), 4-chloroaniline (CLA), 1-(3,4-dichlorophenyl)-3-methylurea (DCPMU), 1-(3,4-dichlorophenyl)urea (DCPU), 1-(4-chlorophenyl)urea (CPU) and monuron, this latter considered both as a metabolite and parent compound. The Koc values of the examined substances were determined by the HPLC screening methods, according to the OECD TG 121. DT50 determinations and aged column leaching experiments were carried out according to SETAC procedures. RESULTS AND DISCUSSION: The examined compounds showed a rather wide range of persistence in soil, with DT50 values less than 2 days for DCA and CLA, close to 8 days for DCPU and CPU and from 16 (diuron) up to 24.8 (DCPMU) days for the others. Their mobility was generally high, based on their Koc values, which ranged from 33 (CPU) to 406 (linuron). The GUS indices indicated that monuron has a clear potential to contaminate groundwater (> 2.8); DCPMU, monolinuron, CPU and diuron are intermediate contaminants (1.8-2.8). Linuron, DCPU, CLA and DCA exhibited a non-leaching behaviour (< 1.8). The aged leaching column experiments showed that parents were found in the leachates at very high percentages respect to the doses applied. The metabolites reached much less percentages, the highest values were observed for monuron from diuron (5.7), CPU (7.2) and DCPMU (8.2%). CONCLUSION: Diuron, Monuron, CPU and DCPMU on the basis of their intrinsic properties, formation from their parents and occurrence in leachates from aged column leaching studies, seem to possess the characteristics of groundwater contaminants. The methodological approach of this study is relatively easy and rapid, hence it can represent a tool for a first screening of compounds such as pesticide metabolites (generally available only in small quantities and for which a field study is not conceivable) or other compounds for which not adequate environmental data are available.     

铜绿假单胞菌降解菲的产物鉴定及代谢过程

研究了1株能快速降解疏水性化合物的铜绿假单胞菌NY3对菲的降解机理。GC-MS分析表明,该菌代谢菲的各个阶段,能够被检测到的主要酸性产物有1-羟基-2-萘甲酸、α-萘酚、邻羟基苯基羰基乙酸、水杨酸、苯甲酸等。铜绿假单胞菌NY3经过一系列氧化代谢将菲转化为1-羟基-2-萘甲酸,进一步反应生成α-萘酚。由菲到1-羟基-2-萘甲酸,未检测到其他产物,说明该阶段产物被继续代谢的速度快,产物含量低不易积累。1-羟基-2-萘甲酸为体系中积累的唯一的主要代谢产物。尽管继续代谢速度慢,但它可以按照α-萘酚、邻羟基苯基羰基乙酸、水杨酸、苯甲酸途径被继续转化。NY3降解菲的代谢产物中未检测到二羟基萘、苯二酚等多羟基化合物,但羟基和羧基处于邻位的化合物,如1-羟基-2-萘甲酸和水杨酸有非常明显积累的趋势,α-萘酚和苯甲酸积累速度次之。     

Cultured amniotic fluid cells for prenatal diagnosis of lysosomal storage disorders: A methodological study

The influence of culture conditions on the ultrastructure and enzyme activities of amniotic fluid cells are reported. Morphological changes were determined as a function of the number of lysosomal-like inclusion bodies per cell, and these results correlated to the activity of Thiexosaminidase, a-mannosidase, β-glucuronidase, arylsulphatase C and 5′ nucleotidase. The parameters examined were pH of the culture media, type of media, increasing cell passage and day of harvest. Our results indicate that enzyme activities are less sensitive to changes in culture conditions as compared to ultrastructural changes. We therefore recommend that in order to obtain reliable ultrastructural results for the diagnosis of storage disorders, cultures should be grown in MEM as the culture medium, the pH of the medium carefully monitored to remain below pH 7·4, examining the cultures no later than the eighth cell passage and no later than the 10th day after subculture.     

Biamnial elevated alpha-fetoprotein and positive acetylcholinesterase in twins,one with anencephaly

Anencephaly in twin B was accompanied by elevated amniotic fluid alpha-fetoprotein (AFP) and a positive acetylcholinesterase (AChE) band on gel electrophoresis in both twin sacs, although twin A was normal. AChE results did not help distinguish the false positive AFP in this set of twins, implying that AChE may diffuse transamniotically as has been previously postulated for AFP. In light of the low concordance rate for neural tube defects in twins, patient counselling in this situation must include the information that AFP and AChE may be falsely elevated in normal twin when the other twin has a neural tube defect.     

The synaptic membrane D2-protein in amniotic fluid from pregnancies with fetal neural tube defects

The D2-protein is a neuronal membrane protein which has an immunochemically detectable soluble derivative in cerebrospinal fluid. We used rocket-on-line immunoelectrophoresis with an antiserum against rat synaptosomal membranes for demonstration of the human D2-protein in fetal cerebrospinal fluid and in amniotic fluids from pregnancies with fetal neural tube defects. D2 was neither found in normal amniotic fluids nor in cases of amniotic fluids from pregnancies with other malformations. Analysis of D2 in amniotic fluids may thus have a future role in the antenatal diagnosis of fetal neural tube defects.     

Separation of amniotic fluid cell types in primary culture by percoll density gradient centrifugation

Amniotic fluid cells obtained by transabdominal amniocentesis at 15–17 weeks of gestation and cultured for 15 and 21 days were separated into three fractions by density gradient centrifugation in Percoll. Each fraction (or peak) corresponded to the following densities: peak A, 1·02–1·03 g/ml; peak B, 1·04–1·05; peak C, 1·05–1·06. Peak A was composed of both non-viable and viable cells; the latter adopted the morphology of epithelial cells in culture. Peak B contained a mixture of fibroblasts and epithelioid cells and peak C had only epithelioid cells. The variability of N-acetyl-β-hexosaminidase and alkaline phosphatase activities was reduced in peaks B and C as compared to that of peak A and of unseparated cells suggesting that more defined and homogeneous cell types for enzymatic determinations can be obtained by centrifugation in Percoll density gradient. In prenatal diagnosis of biochemical defects, the separation of cells would permit a more precise diagnosis by eliminating enzyme variability due to the presence of different cell types or non-viable cells.     

S-100 protein in amniotic fluid of anencephalic fetuses

S-100 protein, which is found essentially in the astrocytes of the nervous system, was assayed in amniotic fluids by Particle Counting Immuno Assay. It was present in 19 cases of anencephaly out of 26, in 1 case of open spina bifida out of 5 and in each of the 4 cases of fetal death, whereas it was not detected in the 48 control amniotic fluids collected between the 16th and the 35th week of gestation. Thirty-one amniotic fluids from fetuses with other congenital malformations were devoid of detectable S-100. The presence of S-100 in amniotic fluid of anencephalic fetuses can presumably be considered as a biological sign of necrosis of the exencephalic brain and seems specific to damage of the central nervous system accompanied by neural tube defect.     

Prenatal diagnosis of non-ketotic hyperglycinemia

We describe successful prenatal diagnosis in four pregnancies at risk for non-ketotic hyperglycinemia, two affected and two unaffected, using the glycine level and the glycine/ serine ratio in amniotic fluid obtained at 16 weeks gestational age. Although this method of prenatal diagnosis for non-ketotic hyperglycinemia has been effective in our hands the narrow differences between affected and unaffected pregnancies indicate the need for caution concerning its reliability.     

Prenatal diagnosis of atypical tay-sachs disease by chorionic villi sampling

We studied a family at risk for atypical TSD in which the index case showed, clinically, a late onset and a gradual psychomotor deterioration and biochemically, a residual hex. A activity in leucocytes. Two prenatal diagnoses of affected fetuses were made in this family, The first one on amniotic cells, the second one on trophoblast biopsy samples. Both of them were confirmed after abortion on cultured cells. Prenatal diagnosis of TSD, even of some atypical forms is possible using trophoblast biopsy, but formal confirmation should be obtained on cultured trophoblasts.     

Chemical studies on Indopacific Ceratosoma nudibranchs illuminate the protective role of their dorsal horn

Summary. Two Ceratosoma species, C. trilobatum and C. gracillimum (Mollusca: Gastropoda: Opistobranchia), collected along the South China Sea coast, have been studied in order to investigate their defensive strategies. Both species contain four typical sponge furanosesquiterpenoids, probably with a dietary origin. The tissue distribution of these compounds has been studied by ¹H-NMR analysis and preliminary bioassays have been carried out in order to evaluate the ecological role of the isolated metabolites. The main sesquiterpene component in both species was (–)-furodysinin (2), which showed significant feeding-deterrent and ichthyotoxic properties in bioassays with freshwater organisms. The work has been performed studying a very limited number of molluscs that prevented the realization of ecologically relevant experiments. However, the almost exclusive presence of compound 2 in the crude extract of the mantle glands concentrated in the dorsal horn of C. gracillimum strongly supports the defensive role of that dorsal protuberance, which probably acts as a defensive lure attracting potential predators to the part of the animal that contains the distasteful chemicals.