Melanoma epidemic across the millennium: time trends of cutaneous melanoma in Emilia-Romagna (Italy) from 1997 to 2004.

BACKGROUND: After a rapid increase in malignant melanoma (MM) incidence in the last decades, trends of the melanoma epidemic in the recent years seemed not homogeneous. OBJECTIVE: This study aimed at the monitoring of some epidemiological data referring to melanoma in a region of the Northern Italy during the past 8-year period. METHODS: All cases of melanoma, including also in situ lesions, diagnosed in Emilia-Romagna and San Marino State, with the exclusion of Cesena province, from 1997 to 2004 were recorded and the incidence of melanoma, adjusted for the European standard population by the direct method, was calculated. RESULTS: Mean standardized incidence was 9.7 for invasive MMs and 11.9, considering also in situ ones, showing an ascending trend with an increment of 3.3 new incident cases in 2004 compared with 1997. No differences in age distribution, gender and site were reported. Concerning tumour thickness, although a general ascending trend in all subtypes, only thin melanoma incidence significantly increased over the study period. CONCLUSIONS: Contrary to data from Northern European countries, melanoma incidence still showed an ascending trend in the Italian population of Emilia Romagna.

Study on the toxicity of phenolic and phenoxy herbicides using the submitochondrial particle assay.

A simple and rapid in vitro toxicological assay, utilizing submitochondrial particles (SMP), has been used to evaluate the toxic effects of fifteen herbicides belonging to the phenol and phenoxyalkanoic acid chemical classes. The SMP assay allows the quantitative evaluation of the toxicity of compounds with different mechanisms of action: uncouplers, inhibitors of the enzyme complexes involved in reverse electron transfer and in oxidative phosphorylation and chemicals that alter the membrane structure. The two groups of herbicides showed different levels of toxicity. For phenol derivatives, EC50 values ranged from 0.16 microM (ioxynil) to 6.7 microM (2,4-dinitrophenol), whereas for phenoxy herbicides EC50 values ranged from 21 microM (2,4,5-trichlorophenoxyacetic acid, 2,4,5-T) to 110 microM (4-chloro-2-methylphenoxyacetic acid, MCPA). On the average, the toxicity of phenolic compounds is greater than that of phenoxyalkanoic acids by two orders of magnitude. Quantitative structure-activity relationships (QSAR) were developed between EC50 values and various molecular descriptors. The results suggest the existence of different mechanisms of action for the two classes of compounds. The findings obtained for phenolic herbicides are consistent with a protonophoric uncoupling mechanism, whereas for phenoxy herbicides a non-specific mode of action at membrane level can be hypothesized.

Sensitivity of embryotoxicity test with Mytilus galloprovincialis (Lmk) towards some compounds of environmental interest (copper and pesticides).

Some compounds of environmental interest were investigated using the toxicity test with early life stage of bivalve molluscs. Three pesticides (Carbofuran, Atrazine and Malathion), for which literature data are available only for some species of oysters, were tested with Mytilus galloprovincialis (Lmk). The toxicity of copper was investigated both with M. galloprovincialis and with Crassostrea gigas (Thunberg). Bioassays highlighted a good sensitivity towards the compounds, with EC50 values in the order of mg l(-1) for the pesticides and of microg l(-1) for copper. Comparison with other toxicity tests for aquatic environments showed that embryotoxicity test with bivalve is slightly more sensitive than embryotoxicity test with the sea urchin Paracentrotus lividus (Lmk), remarkably more sensitive than Microtox test but less sensitive than tests with the copepod Tigriopus brevicornis.

Temporal and spatial changes of macroalgae and phytoplankton in a Mediterranean coastal area: the Venice lagoon as a case study.

Since the late 1980s the lagoon of Venice, a shallow Mediterranean coastal area, has experienced strong environmental changes. Macroalgae, which were the predominant primary producers of the lagoon, reduced markedly, but neither phytoplankton nor seagrasses replaced them. Temporal and spatial changes in macroalgal standing crop (SC) and phytoplankton concentration were investigated between 1987 and 1998. Maps of macroalgal SC show a marked declining trend. Biomass in fresh weight decreased from: 558 ktonnes in 1987, to 85 ktonnes in 1993 and to 8.7 ktonnes in 1998. As a whole, the biomass in 1998 was only 1.6% of the biomass recorded in 1987. Similarly the macroalgal net (NPP) and gross (GPP) primary production decreased from ca. 1502 and 9721 ktonnes year(-1) to ca. 44 and 229 ktonnes year(-1), respectively. In the early 1990s the clam Tapes philippinarum Adams & Reeve and seagrasses, especially Zostera marina Linnaeus, colonised the bottoms free of macroalgae, but the development of intense clam-fishing activities prevented both phytoplankton blooms and seagrass spreading. Maps of chlorophyll a drawn according to data collected in parallel to macroalgal standing crop show unchanged concentrations. Macroalgae changes are enhanced by comparing annual trends in four areas of the central lagoon during 1989-1992 and 1998-1999. In those areas phytoplankton also decreased significantly. Marked changes of some environmental variables strongly associated with the primary production were recorded both during the lagoon mapping and in the areas studied on a yearly basis.

Benthic macrofauna changes in areas of Venice lagoon populated by seagrasses or seaweeds.

Two areas of the Venice lagoon populated by seagrasses (three stations covered by Cymodocea nodosa (Ucria) Asherson, Zostera marina Linnaeus, Zostera noltii Hornemann) or seaweeds (two stations: one covered by Ulva rigida C. Agardh and another at present without seaweed biomass) were monitored by means of six surveys over a year in order to study macrofaunal composition and seasonal changes. The seagrass stations showed a mean species richness (28-30 S m(-2)), individual abundance (1854-4018 N m(-2)) and biomass (22.3-37.7 g m(-2) ash-free-dry-weight, AFDW) ca. 3-8 times higher than those populated by seaweeds (10-15 S m(-2), 494-1395 N m(-2) and 5.6-13.7 g m(-2) AFDW). Differences among seagrass or seaweed stations were much lower. The Ulva-dominated station showed a macrofauna completely different both from the other stations and the communities recorded ca. 30 years ago, before the prolific growth of Ulva. In this station, frequent biomass decompositions and anoxic crises created critical conditions for life favouring organisms with reduced life cycles, younger individuals and the epifaunal species instead of the infaunal ones. In particular, Ulva grazers and scrapers such as Gammarus aequicauda Stock and Gibbula adriatica Philippi were found to be by far the most abundant species, whereas the taxa characteristic of the associations found in the past, in the presence of seagrasses or seaweeds and typical of low eutrophicated environments, appear strongly reduced. Marked differences in the macrophyte dominance and in the bio-physico-chemical variables which characterise the main environmental conditions of the Venice lagoon support the different distribution and composition of macrofaunal communities. Seaweed stations appear mainly governed by the seasonal cycles of these un-rooted macrophytes which, by alternating periods of production and decomposition, are responsible for the drastic reduction of macrofauna biodiversity and biomass. Conversely, seagrass stations exhibit a better oxidisation of the environment and show conditions more favourable for macrofauna colonisation, especially in the presence of macrophytes which are characterised by very well developed below-ground systems such as Cymodocea nodosa.

Sperm cell toxicity test using sea urchin Paracentrotus lividus lamarck (Echinodermata: Echinoidea): sensitivity and discriminatory ability toward anionic and nonionic surfactants.

A reliable sperm cell toxicity test procedure has been developed for the Mediterranean sea urchin Paracentrotus lividus. The sensitivity and discriminatory ability of the test were investigated with regard to surfactants and their biotransformation products. Aromatic and aliphatic surfactants of anionic (linear alkylbenzene sulfonates [LAS]) and nonionic (alcohol polyethoxylates [AE] and nonylphenol polyethoxylates [NPE]) types and their aerobic biodegradation products, i.e., sulfophenylcarboxylates (SPC), polyethylene glycols (PEG), carboxylated polyethylene glycols (PEGC), carboxylated AE (AEC), and nonylphenol (NP), were examined in order to elucidate the influence of their molecular structure on toxicity. Experimental results reveal that the sperm cell test showed good discriminatory ability among all tested compounds, median effective concentration (EC50) values differing by about four orders of magnitude. The toxicity of anionic surfactants depends on the length of the alkyl chain and that of nonionic surfactants is due to their length and branching. Much lower toxicity was shown by aerobic biodegradation products in comparison with that of their parent compounds, with the exception of NP. The obtained EC50s were comparable with available literature data and constitute new toxicity data regarding surfactants for sea urchins.

Solid-state investigation of fluocinolone acetonide.

Three crystalline modifications of fluocinolone acetonide, A, B and C, were characterized by means of FTIR, DSC, TG-FTIR, MICRO-FTIR and X-ray diffractometry. They were easily differentiated by their IR absorption bands in the 3600-3400 cm-1 range. The thermal behavior was also elucidated using combined techniques; thermomicroscopy and thermogravimetry coupled with Fourier transform infrared spectroscopy were found to be very useful tools for a better understanding of thermal events. On heating, form A and C were fully converted into form B. Polymorph A was found to be enantiotropically related to B, while form C was monotropically related to B. The existence of three polymorphs was confirmed by means of their different X-ray diffraction patterns. Detailed methods of preparation of the three modifications are also described.

Chromosomal fragility in the cancer-prone disease xeroderma pigmentosum preferentially involves bands relevant for cutaneous carcinogenesis.

Spontaneous and folate-induced chromosomal fragility was analyzed in peripheral blood lymphocytes from 6 patients affected by the cancer-prone disease xeroderma pigmentosum (XP), from the parents of 4 of the patients, and from 12 normal subjects. All XP patients were defective in nucleotide-excision repair; 4 belonged to group C and 1 each to groups A and D. A tendency toward increased spontaneous chromosomal fragility was observed in the XP family members, and lesions indicating substantial chromosomal damage, which were not observed in any healthy donors, were frequently found. The spontaneous lesion sites in lymphocytes from homozygous and heterozygous carriers of XP defects appeared to be significantly associated with those observed in normal skin fibroblasts from the same subjects. These XP spontaneous fragility sites showed a statistically significant association with the rearrangement breakpoints reported in skin pre-tumoral and tumoral lesions from normal and unrelated XP donors. Under conditions of folate deprivation, the chromosomal fragility level, the pattern and the frequency of expression of fragile sites in XP patients and in their parents were similar to normal. However, XP patients generally showed a higher susceptibility to breakage at sites described as mutagen and carcinogen targets.

Lack of major mitochondrial bioenergetic changes in cultured skin fibroblasts from aged individuals.

Human skin fibroblasts from young and old donors were cultured in vitro and compared in their mitochondrial morphology and function. A decreased numerical density of mitochondria in the fibroblasts from old individuals was balanced by the increased size of individual mitochondria. The mitochondrial membrane potential, estimated in the intact cells by the difference between the total accumulation ratio of the lipophilic cation tetraphenylphosphonium and the accumulation ratio in presence of uncoupler, was unchanged, as were some mitochondrial enzymatic activities tested in the homogenates. The results point out that the decreased proliferating capacity observed in the fibroblasts from the old subjects was accompanied by a likely decrease of mitochondrial duplication; the decreased energy utilization for cell division balances a possible energetic decline in such way that the steady-state energy status is unchanged.

Dyschromatosis symmetrica hereditaria associated with idiopathic torsion dystonia. A case report.

The authors report a case of a family in which a diagnosis of dyschromatosis symmetrica hereditaria was established by a clinical pattern of cutaneous lesions and by assessment of cellular DNA repair synthesis. The skin lesions were characterized by a mixture of hyperpigmented and hypopigmented macules and were localized on the back of the feet of three patients (two male brothers and one sister). All the patients also had small freckle-like pigmented macules on their face. The father presented large symmetrical hypopigmented vitiligo-like macules. In this patient, mottled pigmentation and depigmentation of the extremities had been present since childhood. In the four patients no cellular abnormalities in DNA repair ability were found. These data exclude a mild form of xeroderma pigmentosum. The daughter, a 9-year-old girl, had since the age of 7 also shown a neurological disorder diagnosed as idiopathic torsion dystonia. The authors emphasize the association of dyschromatosis symmetrica hereditaria, a rare cutaneous disease, with idiopathic torsion dystonia, a rare idiopathic neurological disease, in this young girl.

Immune defects in families and patients with xeroderma pigmentosum and trichothiodystrophy.

Xeroderma pigmentosum (XP) is a rare autosomal recessive disease characterized by photosensitivity, a high incidence of cancer in sun-exposed portions of the skin and a reduced capacity to repair the u.v.-induced DNA damage. One of the XP mutations (XP-D) has also been identified in patients affected by trichothiodystrophy (TTD), a rare autosomal recessive disease characterized by brittle hair, mental and physical retardation, peculiar face and ichthyosis. However, in these patients there is no evidence of increased skin tumour incidence. Since an impairment of cell-mediated immunity has been proposed as a co-factor in the cancer proneness of XP patients, we investigated the involvement of immune defect(s) in five XP patients, five TTD patients, their parents, and 24 TTD relatives. We evaluated the phenotype of circulating lymphocytes, natural killer (NK) cell lytic activity, target cell binding of NK cells at single cell level and the effect of interferons (IFN) alpha and beta on NK cell activity. The relative proportion of CD3+ and CD4+ circulating lymphocytes was reduced in XP but not in TTD patients. NK cell lytic activity was decreased in XP patients and their mothers, but their fathers showed normal lytic activity. NK activity varied among TTD families: four out of five patients and their relatives presented low NK cell activity, and one family was normal. In TTD family members, NK activity increased after incubation with IFN-alpha or IFN-beta, but never reached normal values. In contrast, in XP patients and their mothers, the defect was almost completely corrected after in vitro incubation with IFN-alpha or IFN-beta. Our study indicates impaired NK lytic activity in the majority of TTD and XP patients and that this defect is present also in members of their families. In addition, XP patients present a low number of circulating T cells. These multiple abnormalities, together with DNA repair defects, could be related to the increased cancer risk in XP patients.

Trisomy 21 mosaicism in two subjects from two generations.

In the course of a chromosome fragility investigation on the cancer prone hereditary disorder xeroderma pigmentosum, a low proportion of cells with a 47,XY,+21 karyotype was found in lymphocyte cultures of a patient not showing any Down syndrome symptom. The presence of trisomy 21 mosaicism was demonstrated also in peripheral blood of the healthy father and confirmed by "chromosome painting" that allowed a rapid detection of chromosomes 21 on metaphase cells and interphase nuclei. The trisomic cell line was not detected in fibroblast cultures. The analysis of chromosome 21 heteromorphism indicated that in both subjects the mosaic could result from either a diploid or an aneuploid zygote. Since in the trisomic cell line of the father and the son the extra chromosome 21 seems to be the same, a predisposition toward mitotic errors (non-disjunction or anaphase lagging) may be postulated, leading to the recurrent gain or loss of a specific chromosome 21. In order to test the hypothesis of an abnormal mitotic behaviour of the chromosome 21, we investigated the centromere separation index and the DNA restriction pattern in Southern blots probed with satellite DNA sequences specific for chromosome 21 centromere. Both the approaches did not reveal any peculiar feature that may account for the genetically determined proneness to mitotic error observed in the family.

Chromosome rearrangements in normal fibroblasts from xeroderma pigmentosum homozygotes and heterozygotes.

Chromosome analysis was carried out in cultured fibroblasts from unaffected skin of five unrelated xeroderma pigmentosum (XP) patients and nine family members. Structural chromosome changes were observed in cultures from all examined individuals. Furthermore, in one XPD patient and in one XPC patient and his parents, cytogenetically abnormal clones were detected. Some of these clones were present starting from the primary explant. This cytogenetic pattern is similar to that observed in an XPC patient previously studied by us. The analysis of breakpoint distribution from clonal and non-clonal chromosome rearrangements showed that some breakpoints were more frequent and common to different families or to different family members although definite evidence of preferential involvement of chromosome bands was not obtained. This investigation indicates that there is a consistent tendency toward chromosome instability in XP mutation carriers. The instability could be related to the multiple chromosome anomalies characterizing skin tumors in XP subjects.

[Thymopentin in the treatment of recurrent condylomata acuminata]. / La timopentina nel trattamento dei condilomi acuminati recidivanti.

Twelve patients who had been suffering from genital and/or perianal recurrent condyloma acuminatum for a minimum of 5 to a maximum of 24 months, in spite of treatment, were studied from the immunological viewpoint and treated with 50 mg s.c. Thymopentin three times a week for 4 or 6 weeks. Six of the patients were cured at the end of treatment, five after 5 months, and one was not cured. Analysis of the clinico-laboratory data shows a significant agreement between the course of clinical signs and the immunological picture. The various cure stages are probably attributable to the basic immune arrangement which was more impaired in the 5 patients who were cured more slowly and in the non-cured case. In the latter too, however, Thymopentin permitted correcting the balance of the relationship between the various lymphocyte subpopulations.