Nutraceutical value and safety of tomato fruits produced by mycorrhizal plants.

Tomato fruit has assumed the status of 'functional food' due to the association between its consumption and a reduced likelihood of certain types of cancers and CVD. The nutraceutical value of tomatoes can be affected by the cultivation conditions, e.g. the phytochemical content of the fruits may increase with the establishment of beneficial mycorrhizal symbioses in the plants. A multidisciplinary study was carried out to gain knowledge on the antioxidant, oestrogenic/anti-oestrogenic and genotoxic activity of tomato fruits produced by mycorrhizal plants. The present results showed that the symbiosis positively affected the growth and mineral nutrient content of tomato plants and enhanced the nutritional and nutraceutical value of tomato fruits through modifications of plant secondary metabolism, which led to increased levels of lycopene in fruits obtained from mycorrhizal plants, compared with controls. Moreover, such changes did not result in the production of mutagenic compounds, since tomato extracts induced no in vitro genotoxic effects. Fruit extracts, both hydrophilic and the lipophilic fractions, originating from mycorrhizal plants strongly inhibited 17-ß-oestradiol-human oestrogen receptor binding, showing significantly higher anti-oestrogenic power compared with controls. The present study shows that beneficial plant symbionts, such as mycorrhizal fungi, can lead to the production of safe and high-quality food, which is an important societal issue strongly demanded by both consumers and producers.

National pathfinder survey on children's oral health in Italy: pattern and severity of caries disease in 4-year-olds.

This paper describes the dental health status of Italian 4-year-olds in 2004/2005 and analyzes the association between caries in preschool children and some background characteristics in children and parents. Caries was recorded according to WHO criteria. 5,538 subjects were examined. Information on the children's and their parents' social, behavioral, ethnic and demographic status was obtained through a series of closed questions. Bivariate analysis was performed to investigate the association between caries and background characteristics. The probability of being an extra zero for the dmfs index was estimated via the zero-inflated negative binomial regression model (ZINB). 78.4% (95% CI = 77.2-79.6) of the children were caries-free. The national mean dmfs index was 1.36 (95% CI = 1.15-1.57), ranging from 1.22 (95% CI = 1.03-1.42) in the Italian North-East to 1.73 (95% CI = 0.83-2.63) in the South section. Significant bivariate associations between caries experience and risk factors were observed for parents' nationality (not Italian vs. Italian: p < 0.001), parents' educational levels (low vs. high: p < 0.001), preterm birth (yes vs. no: p = 0.011), prolonged breastfeeding (13 months: p = 0.038) and early tooth eruption (<6 months as reference: p = 0.004). Multivariable analysis (ZINB) showed that children with a low caries risk level had a higher probability of being an extra zero; in particular, children from fathers with a high educational level showed a probability of being an extra zero of 0.22. The results suggest a need to plan preventive programs to reduce oral health disparities among Italian preschool children, based on educational intervention with parents and children concerning oral health and caries prevention.

Estradiol-antagonistic activity of phenolic compounds from leguminous plants.

Natural flavonoids are currently receiving much attention because of their estrogenic and antiestrogenic properties. Six isoflavones (isoprunetin, isoprunetin 7-O-beta-D-glucopyranoside, isoprunetin 4',7-di-O-beta-D-glucopyranoside, genistein, genistein 7-O-beta-D-glucopyranoside, daidzein), four flavones (luteolin, luteolin 7-O-beta-D-glucopyranoside, luteolin 4'-O-beta-D-glucopyranoside, licoflavone C), isolated from Genista morisii and G. ephedroides (two Leguminosae plants of the Mediterranean area) together with two structurally related pterocarpans, bitucarpin A and erybraedyn C, isolated from Bituminaria bituminosa (Leguminosae), were tested for the antagonist activity by a yeast based estrogen receptor assay (Saccharomyces cerevisiae RMY326 ER-ERE). Most compounds inhibited the estradiol-induced transcriptional activity in a concentration dependent manner. In particular, for the flavone luteolin 77% inhibition of the induced beta-galactosidase activity was observed. Interestingly, licoflavone C exhibited a dose-dependent antagonistic activity at concentrations up to 10(-4) M, but stimulated beta-galactosidase expression at higher concentrations resulting in a U-shaped-like dose-response curve.

National Pathfinder survey of 12-year-old Children's Oral Health in Italy.

No recent data on the experience of caries among Italian 12-year-olds are available. In 2004, an epidemiological survey called 'National Pathfinder among Children's Oral Health in Italy' was promoted and carried out. This study reports the actual oral health status of Italian 12-year-olds according to gender, residence area and geographical distribution. Clinical examinations were carried out from March 2004 to April 2005, according to WHO criteria, and included dental caries (decay at the dentinal lesion level) and Community Periodontal Index (CPI). 5,342 children (2,670 males, 2,672 females) were examined by 7 ad hoc calibrated raters. Dental caries experience was found in 43.1% (95% CI 41.8-44.4%) of the study population. The mean DMFT score was 1.09 (95% CI 0.98-1.21). Significant differences (p < 0.05) were observed among geographical sections for DT, FT and DMFT. An inverse relationship was observed between mean DMFT and gross national product per capita (p < 0.001). Gingival bleeding was observed in 23.8% of children, while 28.7% had calculus. Significant differences in CPI scores among sections were found throughout the sample in both males and females (p < 0.001). Over the past two decades, mean DMFT fell from over 5 to its present level, halving every decade; consequently, the recorded level of dental caries has become aligned with that in other Western European countries. Nevertheless, differences in DMFT values remain between children from different socioeconomic backgrounds.

Occurrence of estrogen-like substances in the marine environment of the Northern Mediterranean Sea.

Evidence has been presented that a number of environmental agents perturbs the function of the sex hormone signalling pathways in marine animals. In this work the estrogenic/antiestrogenic activity of coastal marine water samples from five sites of the Northern Mediterranean Sea was assayed using a recombinant Saccharomyces cerevisiae yeast strain (RMY326 ER-ERE) transfected with the human estrogen receptor alpha. Extraction of potential EDCs from seawater was performed in columns packed with XAD-2 resins. The estrogenic activity was calculated as percentage of the activity of 17beta-estradiol (10 nM) and it ranged from 4.8% and 59.03%, significantly depending on sampling site (p=0.0013) and season (p<0.05). Antagonistic activity of extracts was also detected and the percentage of inhibition of estradiol-dependent beta-galactosidase induction ranged to 52.8%. These results point the reliability of the yeast assay as a first level screening test to assess the quality of aquatic environments.

Assessment of estrogenic activity of flavonoids from Mediterranean plants using an in vitro short-term test.

Six isoflavones, daidzein (4',7,-dihydroxyisoflavone), genistein (4',5,7-trihydroxyisoflavone), genistin (genistein 7-O-beta-D-glucopyranoside), isoprunetin (4',7-dihydroxy, 5-metoxyisoflavone), isoprunetin 7-O-beta-D-glucopyranoside, isoprunetin 4',7-di-O-beta-D-glucopyranoside and four flavones, luteolin (3',4',5,7-tetrahydroxyflavone), luteolin 7-O-beta-D-glucopyranoside, luteolin 4'-O-beta-D-glucopyranoside, licoflavone C (4',5,7-trihydroxy,8-isoprenylflavone) were purified from Mediterranean plants (Genista morisii and Genista ephedroides) and their estrogenic activity was assessed by a yeast reporter gene assay (Saccharomyces cerevisiae RMY326 ER-ERE). Licoflavone C showed a powerful estrogenic activity at 10(-7) M (0.0338 microg/ml) and it was 47.45% than 10(-8) M 17beta-estradiol (0.00272 microg/ml). The estrogenicity of this flavone was found to be comparable to the activity showed by genistein at 10(-6) M (0.27 microg/ml). This study points out that a glucose substituent in flavones and isoflavones modulates the hormone-like activity in a different way. Isoflavone aglycones showed a more estrogenic activity than the corresponding glucosides. Conversely, the glucosidation made estrogenic the flavone luteolin and the position of substitution differently influenced the estrogenic activity of compounds.

[Mycobacteria in swimming pool water and the meaning of microbiological conventional indicators]. / Presenza di micobatteri in acque di balneazione e predittività degli indicatori microbiologici convenzionali di qualità igienico sanitaria.

Monitoring program of hygienic quality water in twelve public swimming pools was performed. Legally required microbiological indicator parameters of safety for gastrointestinal illness were measured besides the analyses of Pseudomonas spp. and Staphylococcus spp. prevalence, frequency of recovery and number of nontuberculous mycobacteria. We detected positive samples for coliforms at lower rate (29.3%) than Pseudomonas (75.5%), Staphylococcus spp. (46%) and Mycobacteria (59.4%). We pointed out statistically significant correlation (r=0.67 p=0.0001) between Mycobacteria and Pseudomonas so we think that the latter might be a good predictive marker. As 82% of samples had free chlorine residual within the limits stated by Italian Laws, the efficacy of chlorination to prevent risk of infectious diseases transmission by route other than gastroenteric was discussed. A revision of both the sanitary significance of conventional microbial parameters and the related regulations appears necessary.

A recombinant yeast strain as a short term bioassay to assess estrogen-like activity of xenobiotics.

The study investigated the responsiveness of a recombinant Saccharomyces cerevisiae yeast strain to screen estrogen-like chemicals in order to use it as a first level short-term bioassay for environmental preventive medicine purposes. We used the yeast strain RMY326 ER-ERE, containing the human estrogen receptor alpha and a reporter gene lacZ encoding the enzyme b-galactosidase. We tested the natural steroid hormone 17beta-estradiol, diethylstilbestrol, colchicine, some phytoestrogens and pesticides, and organochlorine insecticides. The relative inductive efficiency (RIE) and EC50 values confirmed the sensitivity and specificity of the yeast strain. A statistically significant reproducibility of results in repeated experiments was assessed.

Characterization of 'faecal streptococci' as indicators of faecal pollution and distribution in the environment.

The recent revision of the taxonomy of 'faecal streptococci' prompted us to verify the importance of identifying the species of this group of cocci. During a study carried out to assess the hygienic quality of environmental samples from a variety of sources, we isolated 198 strains named faecal streptococci on the basis of conventional international tests (EVA broth multiple tube test) used for Public Health purposes. The predominant species were Enterococcus faecalis (39%) and Ent. faecium (29%), followed by Ent. durans/hirae, Ent. casseliflavus/gallinarum, Ent. raffinosus, with a different prevalence of the species depending on the source. Eighty-four per cent of isolates were true faecal species. Only one isolate was identified as belonging to the Streptococcus genus. The authors stress the opportunity to identify the species. This may help to clarify the ecological and epidemiological characteristics of intestinal enterococci and streptococci in the environment, in drinking and recreational waters and their meaning as indicators of faecal pollution. All isolates were tested for their susceptibility to some antimicrobial agents widely used in medical therapy and the pattern was compared with the pattern of isolates from clinical specimens.

Prevalence of Listeria monocytogenes and other listerias in Italian-made soft cheeses.

The frequency of L. monocytogenes and other listerias was determined in different types of Italian-made soft cheeses purchased from retail outlets (shops and supermarkets) located in different areas and different towns of central Italy. Of the 164 examined samples, eight proved to be positive for L. monocytogenes (4.9%), seven strains belonged to serotype 1, and one strain to serotype 4. Thirty-six samples were positive for the presence of other listeria species (22%); of these, L. innocua was prevalent (72% of positive samples). The cheeses bought in supermarkets displayed a higher and statistically significant (Fisher's exact test: 0.0016) positivity for Listeria spp. than those sold by the shops, independently of the type of cheese. One particular type of cheese proved to be frequently contaminated (Fis. ex. test: 0.0013). Technical, analytical and epidemiological aspects are discussed.

Antibiotic susceptibility and serotyping of Pseudomonas aeruginosa strains isolated from surface waters, thermomineral waters and clinical specimens.

Different kind of mineral waters used for drinking purposes or hydromineral therapy and surface environmental waters for recreational use were tested for the presence of Pseudomonas aeruginosa. A comparison of antibiotic resistance patterns and serotyping between water strains and P. aeruginosa isolates from different infection sites of hospitalized humans (Pisa General Hospital) was performed. Altogether one hundred and fifty-six strains were characterized. All isolates were most resistant to furans. A very high degree of resistance to amoxicillin, clavulanic acid, ampicillin, chloramphenicol, tetracycline and trimethoprim was pointed out. Resistance to amikacin, cefotaxime and norfloxacin was observed only in strains from clinical specimens. Amikacin was the antibiotic to which 80% of multiple resistant strains showed sensitivity. The distribution of O-antigen serotypes varied by samples sources; 01, 06, 011 serotypes were the most frequently recovered. No isolates were found to be 08, 013, 014, 015 immunotype. The data indicate that serotyping can be a useful addition to antibiotic susceptibility testing for epidemiological purposes.

Preliminary investigation of suitable bacteria strains for the detection of bacteriophages in environmental polluted surface waters.

Different host-specific bacteriophages were monitored in several water systems together with the usual faecal contamination indicator bacteria (thermotolerant coliforms). The host bacteria used for the plaque assay were E. coli K12, E. coli K12 Hfr, E. coli CHfr and S. typhimurium LT2F+. S. typhimurium LT2F+ showed significantly lower phage counts than the three E. coli strains. The phages specific for E. coli K12 (somatic phages) were in high correlation (r = 0.86, r = 0.91; P less than 0.001) with other coliphages, but not with phages specific for S. typ. LT2F+ (r = 0.33; P less than 0.05). There was an high degree of correlation among phages of E. coli K12 Hfr, E. coli C Hfr and S. typhimurium LT2F+ (pilus F-carrying bacteria) (r = 0.96, r = 0.73, r = 0.75; P less than 0.001). Good correlation was seen between faecal coliforms and male-specific phages but not among faecal coliforms and E. coli K12-specific phages (r = 0.30; P less than 0.05). The relative suitability of host strains when using phages in the evaluation of faecal and viral pollution of aquatic environments is discussed.

Serum antibodies to polioviruses in a Tuscan population, Italy.

In order to study the serological status of a population against poliomyelitis, neutralizing antibody for the three types of polioviruses were examined in 742 serum specimens from individuals aged from 1 to 70 years and over were collected during 1985. Sixty-two percent of the sera had antibodies (titre greater than or equal to 1:8) to all three polioviruses, while 5% had no poliovirus antibody at all. However the rates of seropositivity did vary with age: the 10-19 age group had the lowest frequency of antibody (38%) to the three viruses and the difference between the percentage of this group and the previous (1-10 years) and following ones (20-70 years and over) was highly significant. The results of this seroprevalence survey indicate a gap in immunity to poliomyelitis in the population examined.

Human rotavirus detection in stool specimens using enzyme-linked immunosorbent assays and latex agglutination test.

A total of 194 stool samples from sick and healthy children were tested by three commercial tests: two Rotavirus enzyme-linked immunosorbent assays (ELISA) and one latex agglutination test (LTX). The concordance, sensibility and specificity of these tests were calculated. The results suggest to consider as positive the samples reactive to two enzyme immunoassays at least. The latex test, although less sensitive than Enzygnost-Rotavirus and Rotazyme II, was found to be especially useful for the rapid diagnosis of rotavirus gastroenteritis when used to test the first available stool sample from acutely ill patients.

The diagnostic cytology of nonspecific vaginitis.

Diagnostic cytology has not kept pace with the general growing interest in nonspecific vaginitis (NSV). This study on 114 patients compares Papanicolaou vaginal smears with bacteriologic cultures, whiff test, gram-stained vaginal smears, and vaginal pH in the evaluation of nonspecific vaginitis. Cytology is a valid practical test for diagnosis for nonspecific vaginitis; however, it must be emphasized that the coccoid morphotype is the same as the Gardnerella morphotype. Perhaps the Gardnerella morphotype is the most typical of all the inflammatory typologies found in vaginal smears, and it was indeed one of the first to be described in the history of diagnostic cytology--albeit with a slight misnomer. The question is of considerable importance since NSV is the most frequent and persistent of all collapses of the vaginal ecosystem. Cases of NSV must be screened to prevent more serious diseases of the female genital tract.