Metals in plasma of nonagenarians and centenarians living in a key area of longevity.

The concentration of calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn) and selenium (Se) in plasma of 76 nonagenarians (mean age, 89.0±6.3 years), 64 centenarians (mean age, 101±1 years) and 24 middle-aged subjects as controls (mean age 61.2±1.1 years), was determined by sector field inductively coupled plasma mass spectrometry. All the subjects lived in Sardinia, an Italian island, that has the higher prevalence of centenarians than in other European countries. A comparison among the three classes of age showed a significant depletion of Ca, Co, Fe, Mn and Se (all p<0.001) in nonagenarians and centenarians with respect to controls. In particular, the geometric mean (GM) values of Ca, Co, Fe, Mn and Se were: 94.1 µg/ml, 0.46 ng/ml, 1314 ng/ml, 2.47 ng/ml and 111 ng/ml in controls; 87.6 µg/ml, 0.22 ng/ml, 815 ng/ml, 1.07 ng/ml and 88.9 ng/ml in nonagenarians; 87.0 µg/ml, 0.29 ng/ml, 713 ng/ml, 1.27 ng/ml and 81.9 ng/ml in centenarians. The highest inverse relationship with age was observed for Fe (p<0.001; ρ=-0.352) and Se (p<0.001; ρ=-0.417). This trend was also observed when data were sorted by gender. On the other hand, Cu and Mg levels in plasma remained substantially unchanged during aging. As regards Cu, it was significantly higher in females than in males in controls (GM, 1294 ng/ml vs. 1077 ng/ml; p=0.012), in nonagenarians (GM, 1216 ng/ml vs. 1081 ng/ml; p=0.011) as well as in centenarians (GM, 1226 ng/ml vs. 1152 ng/ml; p=0.045) and in hypertensive subjects with respect to healthy people (GM, 1215 ng/ml vs. 1129 ng/ml; p=0.021). These data can be used to enhance knowledge and support the research on: i) metals involved in aging in areas with high rates of human longevity; ii) variables (gender, lifestyle habits and health status) as critical determinants in aging; and iii) mineral intake and supplementation at older age affecting the healthy aging.

Cadmium influences the 5-Fluorouracil cytotoxic effects on breast cancer cells.

The aim of the research was to evaluate a heavy metal, Cadmium (Cd), which was used to produce alterations in human breast cancer cell line MCF-7. Moreover, we analyzed both immunohistochemical and ultrastructural alterations induced by the antineoplastic drug, 5-Fluorouracil (5-FU), after exposure to different concentrations of Cadmium. Also, we compared the effects of these compounds on actin and tubulin cytoskeleton proteins. Under ultramicroscopic observation, control cells looked polymorphous with filopodia. In cells already treated with small concentrations of Cd, after brief times of incubation, we observed an intense metabolic activity with larger, clearer, and elongated mitochondria characterized by thin and numerous dilated cristae. 5-FU-treated cells showed cytotoxicity signs with presence of pore-like alterations in the cell membrane and evident degeneration of cytoplasm and cell nuclei. The addition of 5-FU (1.5 µM) to the cells treated with Cd (5 µM-20 µM) did not induce significant ultrastructural changes in comparison with cells treated only with Cd. In Cd+5FU-treated cells mitochondria with globular aspect and regular cristae indicated the active metabolic state. In cells treated only with Cd we observed alterations in actin distribution, while tubulin branched out throughout the cytoplasm. With the association of Cd+5FU, we observed less morphological alterations in both tubulin and actin cytoskeleton proteins. Although the mechanism remains unknown at present, our findings suggest that Cd prevents the cytotoxic effect of 5-FU on breast cancer cells. These preliminary results could have an important clinical application in patients with breast cancer.

Low cadmium concentration in whole blood from residents of northern Sardinia (Italy) with special reference to smoking habits.

INTRODUCTION: The present study was initiated to investigate the cadmium concentrations in whole blood of Northern Sardinian, non-occupationally exposed adult subjects. Sardinia is a large Italian island which differs genetically and environmentally from other mainland Italian areas. METHODS: Two hundred and forty-three adults (157 females and 86 males) were selected in the study area from subjects who were undergoing blood collection for laboratory analysis during the period January 2005-May 2005. Whole blood was analysed by graphite furnace atomic absorption spectrometer equipped with a Zeeman-effect background corrector (Perkin Elmer ZLS5100) and an auto sampler. The adopted analytical procedure uses the Stabilized Platform Temperature Furnace (STPF) technique. RESULTS: The mean value of Blood Cadmium Concentration (BCdC), expressed as Geometric Mean, was 0.32 pg/l (CI 95%: 0.31-0.34 l) in non-smokers to 034 pg/l (CI 95%: 0.30-0.39 pg/l) in ex-smokers up to 0.47 gg/ll(CI 95%: 0.42-0.53 pg/l) in smokers (p < 0.0001). DISCUSSION: The results show that BCdC levels in Northern Sardinian non-occupationally exposed adults are lower than levels found in many other regions, including those within Italy. Nevertheless, similar values have been detected in other European countries and cities. CONCLUSIONS: In relation to other reports in which data were analysed by strata for smoking habit and age, we found similar BCdC values among non smokers. However, Sardinian smokers seem to show lower levels of blood cadmium.

Contractile regulatory proteins tropomyosin and troponin-T as indicators of the modulatory role of retinoic acid.

Retinoic acid (RA), the active metabolite of vitamin A, plays a significant role in regulating cardiac form and function throughout the life of the organism. Both cardiac morphogenesis and myocardial differentiation are affected by alterations in RA homeostasis. In order to test the effect of all-trans RA and 13-cis RA on cardiomyocyte differentiation, we studied the level and the subcellular compartmentalization of alpha-tropomyosin and troponin-T proteins in cultures of chick embryo cardiomyocytes obtained from Hamburger and Hamilton's (HH) stage 22, 32 and 40 embryos. The retinoids increased the levels of alpha-tropomyosin and troponin-T in the cytoplasmic and cytoskeletal fractions of cells at all three stages of development. The greatest increases in alpha-tropomyosin occurred in the cytoplasmic fraction in HH22 cells cultured for 24 h with all-trans RA or 13-cis RA, whereas the greatest increases in troponin-T were found in the cytoplasmic fraction of HH32 cells exposed to retinoids for 24 h. In cultures treated for 48 h with retinoids, the levels of alpha-tropomyosin and troponin-T showed significant increases in the cytoplasmic compartment of cells treated in HH32-with respect to the control values. These findings are further evidence that RA plays a modulating role in the formation and reorganization of sarcomeric proteins during the process of cardiomyocyte maturation.

Modulation of HLA class I expression in multidrug-resistant human rhabdomyosarcoma cells.

An abnormal HLA expression has been detected in some tumors including rhabdomyosarcoma (RMS). Classical cytotoxic treatment of these tumors, the most common childhood soft tissue malignancy, may induce multidrug resistance (MDR) associated with the expression of a 170-kDa membrane-associated glycoprotein (P-glycoprotein). In order to analyse the connection between modulation of HLA expression and the development of the MDR phenotype mediated by P-glycoprotein in RMS, we used three resistant RMS cell lines; two of these resistant cell lines (TE.32.7.DAC and RD-DAC) were established by in vitro exposure to actinomycin D, a drug of choice in the treatment of RMS; the resistant RMS- GR cell line was established from an embryonal RMS tumor after polychemotherapy. Our results showed that all the resistant cell lines showed a significant increase in the expression of HLA class I surface antigens in comparison to drug-sensitive cells. Blockade of P-glycoprotein with verapamil led to a decrease in HLA class I expression in RMS resistant cell lines. However, no modulation of HLA class II expression was observed in any of the three analyzed cell lines. These findings support the hypothesis that the development of resistance mediated by mdr 1/P-glycoprotein, directly influences the expression of HLA class I in RMS cells, inducing to upregulation. This effect may be relevant to the application in RMS of immunotherapy against tumor-associated antigens presented by HLA class I molecules.

Peopling of three Mediterranean islands (Corsica, Sardinia, and Sicily) inferred by Y-chromosome biallelic variability.

An informative set of biallelic polymorphisms was used to study the structure of Y-chromosome variability in a sample from the Mediterranean islands of Corsica and Sicily, and compared with data on Sardinia to gain insights into the ethnogenesis of these island populations. The results were interpreted in a broader Mediterranean context by including in the analysis neighboring populations previously studied with the same methodology. All samples studied were enclosed in the comparable spectrum of European Y-chromosome variability. Pronounced differences were observed between the islands as well as in the percentages of haplotypes previously shown to have distinctive patterns of continental phylogeography. Approximately 60% of the Sicilian haplotypes are also prevalent in Southern Italy and Greece. Conversely, the Corsican sample had elevated levels of alternative haplotypes common in Northern Italy. Sardinia showed a haplotype ratio similar to that observed in Corsica, but with a remarkable difference in the presence of a lineage defined by marker M26, which approaches 35% in Sardinia but seems absent in Corsica. Although geographically adjacent, the data suggest different colonization histories and a minimal amount of recent gene flow between them. Our results identify possible ancestral continental sources of the various island populations and underscore the influence of founder effect and genetic drift. The Y-chromosome data are consistent with comparable mtDNA data at the RFLP haplogroup level of resolution, as well as linguistic and historic knowledge.

Overexpression and activation of hepatocyte growth factor/scatter factor in human non-small-cell lung carcinomas.

Hepatocyte growth factor/scatter factor (HGF/SF) stimulates the invasive growth of epithelial cells via the c-MET oncogene-encoded receptor. In normal lung, both the receptor and the ligand are detected, and the latter is known to be a mitogenic and a motogenic factor for both cultured bronchial epithelial cells and non-small-cell carcinoma lines. Here, ligand and receptor expression was examined in 42 samples of primary human non-small-cell lung carcinoma of different histotype. Each carcinoma sample was compared with adjacent normal lung tissue. The Met/HGF receptor was found to be 2 to 10-fold increased in 25% of carcinoma samples (P = 0.0113). The ligand, HGF/SF, was found to be 10 to 100-fold overexpressed in carcinoma samples (P < 0.0001). Notably, while HGF/SF was occasionally detectable and found exclusively as a single-chain inactive precursor in normal tissues, it was constantly in the biologically-active heterodimeric form in carcinomas. Immunohistochemical staining showed homogeneous expression of both the receptor and the ligand in carcinoma samples, whereas staining was barely detectable in their normal counterparts. These data show that HGF/SF is overexpressed and consistently activated in non-small-cell lung carcinomas and may contribute to the invasive growth of lung cancer.

[Incidence and duration of breast feeding in the city of Sassari from 1976 to 1987]. / Incidenza e modalità dell'allattamento al seno nella città di Sassari dal 1976 al 1987.

The authors have studied the incidence and duration of breast feeding (BF) among children from Sassari born in the period 1976 to 1987 (3608 full-term, normal infants). The data have shown a progressive decrease in the incidence and duration of B.F. from 1976 to 1983 followed by a progressive increase in the last 4 years of the study. The authors point out the need for structured incentive program aimed at a better education of the mothers concerning the advantages of breast feeding.

[Height and weight increase in Sassarian children in the last 20 years]. / Incremento staturo-ponderale del bambino sassarese negli ultimi 20 anni.

Height and weight were obtained for 4349 children (2330 males and 2019 females), aged 3-11 years and attending the primary school in Sassari, Sardinian, in 1986. Findings were compared to those reported by De Toni and Aicardi for the same population in 1966 in order to find out possible changes occurred in this lapse of time. The results of our study show that in the last 20 years children have become taller and heavier, but at the same time they seem to be rather "fatter" than in the past.

[Anthropometric study of the Sassari population 6 to 11 years of age]. / Studio antropometrico della popolazione sassarese da 6 a 11 anni.

The average values [+/- SD) for height and weight of 1518 males and 1333 females between 6 and 11 years of age from the city of Sassari (Sardinia) have been determined. Numerical values and smooth curves from the 3rd to the 97th percentile for height and weight have also been calculated. The results were compared with those of the literature and particularly with those published by De Toni et Al. 20 years ago. The data show a significant increase in height and weight among Sardinian children during the past 20 years. The availability of these new curves will permit a more correct evaluation of growth in Sardinian children.

[An anthropometric study of a Sassari population 3 to 6 years of age]. / Studio antropometrico della popolazione sassarese da 3 a 6 anni.

The average values (+/- SD) for height and weight of 812 males and 686 females between 3 and 6 years of age from the city of Sassari (Sardinia) have been determined. Numerical values and smooth curves from the 3rd to the 97th percentile for height and weight have also been calculated. The results were compared with those of the literature and particularly with those published by De Toni et Al. 20 years ago. The data show a significant increase in height and weight among Sardinian children during the past 20 years. The availability of these new curves will permit a more correct evaluation of growth in Sardinian children.

[Immunohistochemical observations on tubulin and alpha actinin during mitosis of cultured fibroblasts]. / Osservazioni immunoistochimiche della tubulina e dell'alpha-actinina durante la mitosi di fibroblasti in cultura.

In this work we studied alpha-actinin and tubulin sites in rabbit fibroblasts in culture. Antibodies anti-alpha-actinin were used for indirect PAP-reaction while antibodies anti-tubulin were used for indirect immunofluorescence method. The observations were carried out by light microscopy, phase-contrast and interference-contrast microscopy, with regard to actinin, and by fluorescence microscopy, with regard to tubulin. During the early mitotic phase, alpha-actinin is localized all over the cell membrane of the fibroblasts, forming a sort strong protective cap, while during diacinesis it forms only rings, localizing below the cell membrane and the philopodia. Thus tubulin forms the bundle fibres during mitotic phases.

[Dynamic aspects of alpha-actinin in cultured rabbit fibroblasts studied with immunofluorescence]. / Aspetti dinamici dell' alpha-actinina in fibroblasti di coniglio in cultura mediante l'immunofluorescenza.

In this work we study alpha-actinin sites in rabbit fibroblasts in culture. For this purpose we extracted alpha-actinin from rabbit striated skeletal muscle and produced the relative antibodies in sheep. Electrophoresis was performed, on PAA slab-gel and the immunodiffusion test on agarose slab-gel. Antibodies were used for direct and indirect immunofluorescence techniques by TRIC and FITC. We noticed that in young fibroblasts alpha-actinin is concentrated in perinuclear regions, while in adult fibroblasts it is scattered, more or less evenly in the cytoplasm and above all on the plates which link the cell membrane to the substractum. Both the direct immunofluorescence method with antibodies anti-alpha-actinin, marked by TRITC, and the indirect one with sheep IgG, marked by FITC, gave identical results.