Vitamin C homeostasis in skeletal muscle cells

In skeletal muscle, vitamin C not only enhances carnitine biosynthesis but also protects cells against ROS generation induced by physical exercise. The ability to take up both ascorbic and dehydroascorbic acid from the extracellular environment, together with the ability to recycle the intracellular vitamin, maintains high cellular stores of ascorbate. In this study, we examined vitamin C transport and recycling, by using the mouse C2C12 and rat L6C5 muscle cell lines, which exhibit different sensitivity to oxidative stress and GSH metabolism. We found that: (1) both cell lines express SVCT2, whereas SVCT1 is expressed at very low levels only in proliferating L6C5 cells; furthermore L6C5 myoblasts are more efficient in ascorbic acid transport than C2C12 myoblasts; (2) C2C12 cells are more efficient in dehydroascorbic acid transport and ascorbyl free radical/dehydroascorbic acid reduction; (3) differentiation is paralleled by decreased ascorbic acid and dehydroascorbic acid transport and reduction and increased ascorbyl free radical reduction; (4) differentiated cells are more responsive to oxidative stress induced by glutathione depletion; indeed, myotubes showed increased SVCT2 expression and thioredoxin reductase-mediated dehydroascorbic acid reduction. From our data, SVCT2 and NADPH-thioredoxin-dependent DHA reduction appears to belong to an inducible system activated in response to oxidative stress.     

JNK/p53 mediated cell death response in K562 exposed to etoposide-ionizing radiation combined treatment

The study of the ability of chemotherapeutic agents and/or ionizing radiation (IR) to induce cell death in tumor cells is essential for setting up new and more efficient therapies against human cancer. Since drug and ionizing radiation resistance is an impediment to successful chemotherapy against cancer, we wanted to check if etoposide/ionizing radiation combined treatment could have a synergic effect to improve cell death in K562, a well-known human erythroleukemia ionizing radiation resistant cell line. In this study, we examined the role played by JNK/SAPK, p53, and mitochondrial pathways in cell death response of K562 cells to etoposide and IR treatment. Our results let us suppose that the induction of cell death, already evident in 15 Gy exposed cells, mainly in 15 Gy plus etoposide, may be mediated by JNK/SAPK pathway. Moreover, p53 is a potential substrate for JNK and may act as a JNK target for etoposide and ionizing radiation. Thus further investigation on these and other molecular mechanisms underlying the cell death response following etoposide and ionizing radiation exposure could be useful to overcome resistance mechanisms in tumor cells.     

A novel VNTR enhancer within the SIRT3 gene, a human homologue of SIR2, is associated with survival at oldest ages

SIR2 genes control life span in model organisms, playing a central role in evolutionarily conserved pathways of aging and longevity. We wanted to verify whether similar effects may act in humans too. First, we searched for variability in the human sirtuin 3 gene (SIRT3) and discovered a VNTR polymorphism (72-bp repeat core) in intron 5. The alleles differed both for the number of repeats and for presence/absence of potential regulatory sites. Second, by transient transfection experiments, we demonstrated that the VNTR region has an allele-specific enhancer activity. Third, by analyzing allele frequencies as a function of age in a sample of 945 individuals (20-106 years), we found that the allele completely lacking enhancer activity is virtually absent in males older than 90 years. Thus the underexpression of a human sirtuin gene seems to be detrimental for longevity as it occurs in model organisms.     

Microbial characteristics of marine sediments in bathing area along Pesaro-Gabicce coast (Italy): a preliminary study

AIMS: This paper evaluates the presence of human pathogen micro-organisms in marine sediments in a coastal area suitable for bathing. In addition, the grain size analysis was performed in order to correlate the characteristics of the sediments and the microbial content. METHODS AND RESULTS: The samples were collected in two small bays along the central Adriatic coast, where breakwaters had been built for the purpose of halting marine erosion. Faecal contamination indicators, Salmonella and Vibrio species, enteric viruses were investigated using standard techniques for isolation and identification. The grain size was determined using calibrated sieves and 'Sedigraph' device. In some samples, the faecal contamination indices overstepped legislative limits. Salmonella was never found. Vibrio and enteric viruses were isolated: the micro-organisms were preferentially abundant in fine sediments. CONCLUSIONs: Marine sediments can represent an important reservoir of allochthonous and marine micro-organisms and the microbial charge correlates with the characteristics of the sediments. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that it is important to analyse marine sediments before defining the quality of coastal areas.     

Chemical composition of European propolis: expected and unexpected results

Ten propolis samples from Bulgaria, Italy and Switzerland were analyzed by GC-MS. As expected, most samples displayed the typical chemical pattern of "poplar" propolis: they contained pinocembrin, pinobanksin and its 3-O-acetate, chrysin, galangin, prenyl esters of caffeic and ferulic acids. Two samples differed significantly: one from the Graubünden Alpine region, Switzerland, rich in phenolic glycerides, and one from Sicily which contained only a limited number of phenolics and was rich in diterpenic acids.     

Effect of Fusarium,Paecilomyces and Trichoderma formulations against aphid Schizaphis graminum

Fungal strains belonging to the genera Fusarium Paecilomyces and Trichoderma were tested in vitro in order to study their effects against Schizaphis graminum one of the major pests of cereal crops around the world. Biological assays were performed using a solid formulation that was obtained from fungal cultures grown on rice and then finely ground (0.2 mm). The occurrence of toxic secondary metabolites (fumonisin B₁ and beauvericin) produced by these fungi was also investigated. In each experiment, three groups of aphids: 15-hour old larvae, 5-day old nymphs with wing buds and wingless morphs were treated with a suspension of a fungal formulation. Some strains belonging to the genera Fusarium and Trichoderma significantly controlled the specimens of the three groups of S. graminum. The F. proliferatum strain ITEM 1407, producing a high level of fumonisin B₁ in the culture (1250 g/g), and F. larvarum strain ITEM 2139 had high insecticidal activity (>60%) within 10 minutes after application. As F. larvarum ITEM 2139 did not produce metabolites toxic to mammals, it might be a good candidate as a biocontrol agent of S. graminum in the field.This revised version was published online in October 2005 with corrections to the Cover Date.     

YKL-40 (cartilage gp-39) induces proliferative events in cultured chondrocytes and synoviocytes and increases glycosaminoglycan synthesis in chondrocytes

YKL-40 (cartilage gp-39), is a mammalian glycoprotein related in sequence to chitinases. Its function is unknown, but it is thought to be involved in tissue remodeling. Immunocytochemical staining of YKL-40 in guinea pig chondrocytes (GPC), rabbit chondrocytes (RC), and rabbit synoviocytes (RS) was higher in dividing cells than in confluent cells, suggesting a participation of YKL-40 in cell cycle events. As assessed by the MTT assay, YKL-40 at 1.9-7.6 nM had dose-dependent mitogenic activity toward the three cell types. At 7.6 nM, YKL-40 increased the number of cells of 42% in GPC, 75% in RC, and 86% in RS after 72 h. YKL-40 also stimulated total proteoglycan synthesis by chondrocytes in a dose-dependent manner as assessed by Na[35SO4] incorporation and cetylpyridinium chloride precipitation. At 9.4 nM, YKL-40 increased proteoglycan synthesis of 42% in GPC and 58% in RC after 24 h. The growth factor properties of YKL-40 may explain the increased tissue remodeling associated with high levels of YKL-40 in joint diseases, and possibly, in malignant pathologies such as breast cancer or colorectal cancer.