Histological changes and micronucleus induction in the zebra mussel Dreissena polymorpha after paraquat exposure.

The herbicide paraquat (PQ), still widely used in developing countries, represents a serious risk factor for human and environmental health. To test the sublethal effects of PQ on the freshwater bivalve Dreissena polymorpha, mussels were exposed to 0.125, 0.250, 0.500 mg/L for 7 and 14 days and histologically screened. PQ's genotoxic potential was also determined in haemocytes by the micronucleus, MN, assay. At concentrations > or = 0.250 mg/L, severe lesions, such as cellular vacuolation, lysis and thinness of the germinative epithelia were observed in the digestive gland and testis. A positive trend between the number of granulocytes and all PQ concentrations was observed in both gonads and digestive glands, addressing the inflammatory capacity of this herbicide on these tissues. Mussels exposed to PQ also exhibited a significant MN induction. The spontaneous MN frequencies ranged from 2.75 to 0.425 per cent, while PQ-induced MN rates in treated mussels were between 3.50 and 1.250 per cent. The histopathological effects on the digestive and reproductive systems, as well as the MN induction in the haemocytes, confirmed the cytotoxic and genotoxic effects of PQ also in D. polymorpha.

Histopathological effects induced by paraquat during Xenopus laevis primary myogenesis.

The oxidative agent paraquat induced tail abnormalities during Xenopus laevis development. Specimens exposed from blastula to the tadpole stage revealed pear-shaped myocytes and irregular intersomitic boundaries. The histological feature of the axial musculature was evaluated in embryos sampled at significant stages of the primary myogenesis. During the somitogenesis PQ-treated embryos showed normal appearing myotomes, but reduced PAS activity in the post-rotating myotomal cells, and myoblasts with slight vacuolations. Once etched from the vitelline envelope, embryos showed severely altered myoblasts with irregular cellular apexes, heavy sarcoplasmic vacuolations, pyknotic nuclei and disorganizing intersomitic boundaries. Myotomes with many necrotic myocytes containing disorganized contractile material and heavily malformed intersomitic boundaries characterized the late myogenic stages. Our results evidence the heaviest PQ histopathological effects to affect myogenesis of post-etched embryos, suggesting a possible linkage between the swimming activity and the oxidative damage to muscle tissue.

OOCYTE DEGENERATION AND ALTERED OVIPOSITORY ACTIVITY INDUCED BY PARAQUAT IN THE FRESHWATER SNAIL PHYSA FONTINALIS (GASTROPODA: PULMONATA).

The freshwater snail Physa fontinalis was used as a bioindicator to study the effects of the herbicide Paraquat (PQ) in laboratory assays. The test solutions used, 0.125, 0.25, and 0.5 mg/l PQ, were in the range of the concentrations recommended for aquatic weed control. The study was carried out in two stages to determine the influence of PQ on the ovipository activity of Physa fontinalis, and the histological effects on these snails. Specimens exposed to PQ continued to be reproductively active, but the number of egg masses and eggs laid decreased significantly. Mortality was almost the same in all the experimental lots, but was significantly related to the production of egg masses only in the controls. The histological analysis showed a clear trend among PQ concentrations and degenerating oocytes, but no visible effects on the male sex-line were observed. By interfering with fertility, PQ has an action that may go well beyond its lethal effect on individuals, suggesting that this herbicide should be strictly regulated in weed control programmes. Moreover, since PQ was observed to interfere with the reproductive process, its endocrine disrupting action must not be excluded.

DDT contamination in Lake Maggiore (N. Italy) and effects on zebra mussel spawning.

Zebra mussel (Dreissena polymorpha) is commonly used as contaminant bioindicator in Europe and North America. We used the zebra mussel to follow DDT pollution trends from 1996 to 1997 in Pallanza bay, Lake Maggiore, near the inlet of the River Toce, after a DDT-manufacturing plant discharging residues into a tributary of the River Toce had closed down. DDT contamination fell off sharply outside the bay, but tissue concentrations of the parent compound and residues remained high in molluscs sampled within the bay a year later. Molluscs collected in Pallanza bay in June 1997 released gametes earlier than those sampled at a nearby reference station. Histological studies showed that a significant percentage of these specimens showed marked oocyte degeneration suggesting that DDTs have endocrine-disrupting effects in this species.

Paraquat embryotoxicity in the Xenopus laevis cleavage phase.

The high Paraquat (PQ, 1-1'-dimethyl-4,4'bipyridylium dichloride) embryotoxicity in Xenopus laevis has been shown to be due to its rapid reduction and instantaneous re-oxidation which produces a reactive oxygen species, ROS. Nevertheless, PQ did not show any effects before hatching, stage 32, which showed a resistance, in early X. laevis development, to oxidative damage. Moreover, in view of its genotoxic properties in several experimental models, we studied PQ in the X. laevis cleavage phase that, characterized by a series of rapid mitotic divisions, might be damaged by genotoxic compounds. Embryos were exposed to 20, 40, 60, and 80 mg/l PQ concentrations from stage 2 to stage 9, and then left to develop in control FETAX solution until stage 47. The 80 mg/l PQ concentration gave 19% embryo mortality at the end of the exposure time, and 16.7% larvae mortality at the end of the test; both values were statistically different from the control, 5 and 6.8% respectively. These results confirmed the high resistance in early X. laevis development to PQ oxidative damage. The malformed larva percentages in the PQ exposed groups were higher as regards the control value but did not show any concentration-response; the most frequent malformed larvae found were affected by abnormal tail flexure coupled with abnormal gut coiling. A further experiment was carried out using the same methodology, but exposing embryos only to the 80 mg/l PQ concentration. The surviving blastulae were embedded in Paraplast, then the slides were stained with 4',6-diamidino-2-phenylindole (DAPI) and the nuclei were examined with a confocal microscope. This new preliminary procedure did not reveal any significant presence of micronucleated micromeres in PQ exposed blastulae with respect to the control. Nevertheless, the mechanism by which PQ induced abnormal tail flexure after cleavage exposure remained unknown. PQ seemed to pass through the jelly coats and vitelline membrane, but it expressed teratogenicity between the 2nd and 3rd day. PQ might be accumulated in the embryos during the exposure, and might express teratogenicity later, but it did not seem to induce genotoxicity during the cleavage phase of X. laevis even at very high concentrations.

Reduction in paraquat embryotoxicity by ascorbic acid in Xenopus laevis.

The toxicity of herbicide Paraquat (PQ, 1-1'-dimethyl-4,4'bipyridylium dichloride) in animal cells is related to its rapid reduction and instantaneous reoxidation to produce the reactive oxygen species. Recently, the PQ evaluation with the Frog Embryo Teratogenesis Assay-Xenopus (FETAX) showed its high embryotoxicity. Supposing that the embryos' death was due to PQ-related oxidative damage, we used ascorbic acid (AA), a well known antioxidant, to reduce the PQ embryotoxicity in Xenopus laevis. Embryos were exposed from stage 8 to 47 to 0.1 mg/l PQ alone, and to PQ with AA concentrations ranging from 20 to 200 mg/l, using the FETAX procedure. PQ caused 72.2% mortality, while 17.1% of surviving larvae were affected by abnormal tail flexure. The PQ mortality percentages were reduced in a clear concentration-response by up to 15.2% in the group exposed to PQ with 200 mg/l AA. The histopathologic diagnoses revealed abnormal notochord flexure coupled with vesiculated, pear-shaped myocytes only in the PQ group. After embryo exposure to PQ with 200 mg/l AA, restoration of normal axial tail structures was evident. In conclusion, PQ embryotoxicity in X. laevis was most likely due to oxidative damage that was drastically reduced by AA.

Paraquat induced embryotoxicity on Xenopus laevis development.

Paraquat (PQ, 1-1'-dimethyl-4,4'-bipyridylium dichloride) is an effective herbicide widely used in agriculture with a rate of application for aquatic weed control ranging from 0.1 to 2 parts per million. Considering its wide-spread presence in Italian wetlands, we studied its embryotoxic effects with the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). The percentage of mortality as well as the percentage of malformed larvae was investigated by probit analysis. The results showed that PQ was highly embryolethal. From a LC(50) of 0.138 mg/l and TC(50) of 0.267 mg/l, a TI(50) of 0.52 was derived; indicating that PQ is to be considered a non-teratogenic compound. Remarkable was the presence of a specific malformation, classified as ventral tail flexure, in the 29% of living larvae exposed to 0.125 mg/l PQ concentration. Their histological examination showed several zones of abnormal somites containing severely affected myocytes. This confirmed the molecular mechanism of PQ toxicity in cell microfilaments. Even at the lowest concentration of 0.0625 mg/l, the difference between the mean head-tail length of control and exposed larvae was statistically significant, a sign of growth retardation. All our data emphasize that PQ must be consider highly embryotoxic on amphibian development, and suggest that this herbicide should be strictly regulated in weed control programs.

Paraquat induces actin assembly in depolymerizing conditions.

The molecular mechanism (or mechanisms) at the basis of paraquat (PQ) (a widely used herbicide) toxicity is far from being fully understood. Until now, two main points of view have emerged: 1) PQ-related cell injuries could be mediated by toxic oxygen free radicals coming from the metabolism of the herbicide by the microsomal enzyme system, and/or 2) PQ, by inducing mitochondrial swelling and breakage, could cause troubles in cell energy charge, then driving the cell to death. Recently, some of cytoskeletal structures (microtubules and microfilaments) have been proposed as further PQ cell targets. The microfilament system in particular seems to be markedly affected by the herbicide, but so far no direct evidence associates PQ to actin damage. In this study, experimental data are presented concerning the direct effect of PQ on actin dynamics in solution. We demonstrate that actin selectively binds PQ; moreover, PQ induces the formation of actin sopramolecular structures in depolymerizing medium (G-buffer). Furthermore, by the interactions with F-actin cross-linking proteins (alpha-actinin and filamin), FITC-phalloidin, and myosin subfragment 1 (S1), it is demonstrated that PQ-induced actin aggregates are undoubtedly built up by F-actin. Electron micrographs showed that PQ-induced actin polymers are very short and tend to aggregate one to another. This mutual cohesion leads to the steric blockage of polymer growing ends as suggested by nucleated actin polymerization assays. Sonication, by releasing F-actin fragments from short polymer aggregates, allows actin polymer ends to regain their growing ability.

Central nervous system involvement secondary to metastatic mixed müllerian tumor of the uterus.

The central nervous system is traditionally considered an uncommon site for metastatic disease from female genital tract tumors. We report the case of a 48-year-old woman with malignant mixed müllerian tumor of the uterus, who developed spinal cord compression by epidural metastasis a few days after the diagnosis of the uterine malignancy. Emergency decompressive laminectomy was performed and a good recovery of the neurological function was achieved. In the following days, while submitted to extensive staging for the uterine malignancy, the patient complained of headache, confusion and visual disturbance. CT scan revealed multiple brain metastases. No other site of metastatic disease could be detected. The patient refused any further treatment and died 1 month later from progressive cerebral disease. Attention should be paid to the possibility of unusual distant metastases associated to uterine sarcoma in order to treat these patients promptly.

Choroid plexus compression of glossopharyngeal nerve in patients with glossopharyngeal neuralgia.

Compression of the glossopharyngeal nerve by the lateral choroid plexus of the 4th ventricle protruding excessively through the foramen of Luschka may be at the basis of some forms of glossopharyngeal neuralgia. This condition was observed in four patients. In one case, an anterior compression of the 9th nerve by a megadolic vertebral artery was also present. The condition was resolved in all patients by separating the nerve from the plexus and transecting it at the level of the foramen of Luschka.

Inhibiting action human glioblastoma extract on haemolytic complement of human and xenogeneic sera.

The action of some extracts of human glioblastoma of the haemolytic complement of human and xenogeneic sera has been examined. The study suggests that the antigenic products of some human glioblastomas may induce an inhibiting effect on the haemolytic complement of human, as well as xenogeneic (rat and rabbit), sera. The inhibiting action on the xenogeneic sera is indicative of the presence of species-specific antigens or tumor cells, comparable to that of normal tissues.

Prediction of the interaction effects Ar+ laser-brain tumours by study of the photoacoustic spectroscopy of cerebral oncotypes.

A preliminary definition of light absorption spectra of brain neoplasm represents a correct approach to the laser therapy of cerebral tumours. The goal should be the identification of those oncotypes more sensive to the laser energy, allowing consequently a dosage of the energy proportional to the light absorption coefficient, reducing any side effects. With this purpose the coefficients of light-absorption of 63 different brain tumours have been analyzed and defined by means of photocoustic spectrometry. The study has been focused at wavelength of the Ar+ laser (4,600-6,000 A). The results show a close relationship between histology, degree of vascularization and/or pigmentation and light absorption: the more immature and/or pigmented oncotypes show the highest coefficients of optical absorption. In the laser therapy of these neoplasms it will be sufficient to use low powers and short exposure time.

Respiratory distress caused by migration of ventriculoperitoneal shunt catheter into the chest cavity: report of a case and review of the literature.

A case is reported in which the peritoneal catheter of a ventriculoperitoneal shunt migrated into the chest cavity causing an hydrothorax. The three similar cases drawn from the literature are reported.

Dose-dependence of doxorubicin effect on actin assembly in vitro.

Doxorubicin (Adriamycin), one of the most potent antibiotics used in tumor chemotherapy, shows many undesirable side effects. We studied the effect of different drug concentrations on the biochemistry of cell motility and, in particular, on potassium-induced actin polymerization. It is well known, in fact, that the actin aggregational status could dramatically influence many cell motility manifestations. Our results clearly show that stoichiometric and substoichiometric amounts of doxorubicin negatively influence actin polymerization by inhibiting both the filament growth and the polymer amount at steady-state; the balance between the two different effects seems to be in relation to the drug concentration. The obtained results could explain some of the doxorubicin effects previously observed in vivo.

Forth ventricle hematoma in a eight-year-old child.

We report the case of a 4th ventricle hematoma due to rupture of an angioma localized in the cerebellar vermis, in an eight-year-old child. Only seven cases of a vascular malformation in such a localization, angiographically and/or pathologically verified, have been previously reported. The girl died, in spite of prompt surgical treatment. Also the clinical presentation of this kind of case is very rare.

Antigenic properties of fresh and lyophylised human dura mater.

The action of human fresh and lyophylised human dura mater on rat, rabbit and human serum complement has been analyzed with the aim of confirming and comparing the species-specific antigenicity of both substances. The results demonstrate that both fresh and lyophylised human dura mater induce a complement consumption on rat and rabbit serum. The phenomenon is related to the species-specific antigenic activity of the tissue, and it is only partially decreased by the lyophylisation procedure. The residual antigenic activity, however, doesn't limit the use of this material as a graft for dural defects repairs in humans.

Evaluation of the Ar+ laser thermal effect in rabbit brain tissue by means of optical absorption coefficients. Photoacoustic measurements.

In predicting Ar+ laser-neural tissue interaction a determination of the optical absorption properties of brain parenchyma appears indispensable. In this study a determination of absorption spectra of different areas of rabbit brain at the wavelength of an Ar+ laser beam is made. The areas considered in the study are frontal and occipital gray and white substance, thalamus, basal ganglia, cerebellar cortex and choroid plexus. Specimens of these areas measuring 2 X 3 X 3 mm were obtained from coronal sections 1 cm thick of 20 rabbit brains. The analysis of optical properties was made by means of a photoacoustic spectrometer which appears the only suitable technique for testing the optical absorption properties of biological materials. The absorption coefficient results were similar in magnitude and constant in all the areas examined, measuring approximately 50 cm-1. The prediction of lack of any selective effect for the different areas is confirmed by the sizes of lesions made in cerebral specimens of the same areas of the contralateral hemisphere by means of Ar+ laser: the lesions increase in size in proportion to the increase in output power and exposure time, independently of the areas considered. Also the histological examination of the areas shows the same morphology to be independent of the variations of both output power and exposure time. The only differences were the presence for short exposure time and low output power of a carbonized layer inner to the layer of coagulative necrosis.

Malignant chemodectoma of the carotid body causing spinal cord compression.

We present a case of a 19-year-old woman with a malignant chemodectoma of the carotid body invading the cervical spine. The interest in the case is due to the rarity both of the lesion and of the spine invasion causing a medullary compression.

Malignant cerebellar astrocytoma. Report of five cases.

Five cases of malignant cerebellar astrocytoma observed during a seven-year period are reported. The analysis of the cases allows us to conclude that malignant cerebellar astrocytoma represents a true tumoral entity quite distinct from cerebellar glioblastoma as well as from medulloblastoma. A perfect correspondence between the histological malignancy and the duration of survival has been noted in all the cases.