The synthetic purine reversine selectively induces cell death of cancer cells

The synthetic purine reversine has been shown to possess a dual activity as it promotes the de‐differentiation of adult cells, including fibroblasts, into stem‐cell‐like progenitors, but it also induces cell growth arrest and ultimately cell death of cancer cells, suggesting its possible application as an anti‐cancer agent. Aim of this study was to investigate the mechanism underneath reversine selectivity in inducing cell death of cancer cells by a comparative analysis of its effects on several tumor cells and normal dermal fibroblasts. We found that reversine is lethal for all cancer cells studied as it induces cell endoreplication, a process that malignant cells cannot effectively oppose due to aberrations in cell cycle checkpoints. On the other hand, normal cells, like dermal fibroblasts, can control reversine activity by blocking the cell cycle, entering a reversible quiescent state. However, they can be induced to become sensitive to the molecule when key cell cycle proteins, e.g., p53, are silenced. J. Cell. Biochem. 113: 3207–3217, 2012. © 2012 Wiley Periodicals, Inc.     

Leptin increases axonal growth cone size in developing mouse cortical neurons by convergent signals inactivating glycogen synthase kinase-3beta

We examined the effects of the adipose hormone leptin on the development of mouse cortical neurons. Treatment of neonatal and adult mice with intraperitoneal leptin (5 mg/kg) induced extracellular signal-regulated kinase (ERK) 1/2 phosphorylation in pyriform and entorhinal cortex neurons. Stimulation of cultured embryonic cortical neurons with leptin evoked Janus kinase 2 and ERK1/2 phosphorylation and activated the downstream effector 90-kDa ribosomal protein S6 kinase. Moreover, leptin elicited the phosphorylation of the phosphatidylinositol 3-kinase effector Akt and evoked Ser-9 phosphorylation of glycogen synthase kinase-3beta (GSK3beta), an event inactivating this kinase. Leptin-mediated GSK3beta phosphorylation was prevented by the MEK/ERK inhibitor PD98059, the phosphatidylinositol 3-kinase inhibitor LY294002, or the protein kinase C inhibitor GF109203X. Exposure of cortical neurons to leptin also induced Ser-41 phosphorylation of the neuronal growth-associated protein GAP-43, an effect prevented by LY294002 and GF109203X but not by PD98059. Ser-41-GAP-43 phosphorylation is usually high in expanding axonal growth cones. Neurons exposed to 100 ng/ml leptin for 72 h displayed reduced rate of growth cone collapse, a shift of growth cone size distribution toward higher values, and a 4-fold increase in mean growth cone surface area compared with control cultures. The leptin-induced growth cone spreading was hampered in cortical neurons from Lepr(db/db) mice lacking functional leptin receptors; it was associated with localized Ser-9-GSK3beta phosphorylation and mimicked by the GSK3beta inhibitor SB216763. At concentrations preventing GSK3beta phosphorylation, PD98059, LY294002, or GF109203X reversed the leptin-induced growth cone surface enlargement. We concluded that the leptin-mediated regulation of growth cone morphogenesis in cortical neurons relies on upstream regulators of GSK3beta activity.     

A fluorescence-based assay for the reductase activity of protein disulfide isomerase

We report on a new spectrofluorimetric assay for the measurement of reductase activity of proteins belonging to the superfamily of thioredoxins such as protein disulfide isomerase (PDI). The assay relies on the preparation of a fluorescence-quenched substrate easily accessible in two steps through functional group transformations of the peptide Gly-Cys-Asp. In the first step fluorescein isothiocyanate is linked to the Gly-NH(2) terminus and in the second step the Cys-SH groups are converted into a disulfide bond. Both intermediate and final substrate have been fully characterized by mass spectrometric and nuclear magnetic resonance measurements. Dimethyl sulfoxide is here reported to be a mild oxidizing agent allowing us to obtain in good overall yield the assay substrate in a single synthetic step. A reliable estimation of PDI reductase activity is obtained via the detection of a strong fluorescence enhancement after enzymatic reduction. Moreover, our assay provides further support for the key role played by thioredoxin reductase in enabling disulfide reductase activity of PDI.     

Kaolin-based particle film technology affects tomato physiology,yield and quality

Environmental stress can affect development and yield of tomato plants. This study was undertaken to investigate the underlying mechanism asserted by kaolin on tomato physiology by evaluating its effect on leaf, canopy and inner fruit temperatures, gas exchange at the leaf and canopy scales, above ground biomass, yield and fruit quality.The study was carried out under field conditions in Southern Italy. Treatments were plants treated with kaolin-based particle film (Surround^® WP) suspension and untreated plants (control).Kaolin application slightly increased leaf and canopy scale temperatures by 1.0 and 0.4 °C, respectively, transpiration rate decreased at both scales. On calm days (wind speed <0.5 m s^?1) with a prevalently clear sky at midday, inner fruit temperature (t_f) of kaolin-treated plants was 4.4 °C lower than the t_f of control plants, while in days with clear sky-windy, and cloudy-calm, the t_f did not differ.At leaf scale, net assimilation was reduced by 26% in kaolin-coated treatments. Stomatal conductance decreased by 53%, resulting in reductions of 34 and 15% in transpiration and internal CO₂ concentration, respectively. Gas exchange parameters measured at canopy scale were similarly affected. In kaolin-treated plants, assimilation and evapotranspiration rates were reduced by 17 and 20%, respectively, while dark respiration was not affected. Above ground dry biomass decreased by 6.4%.Marketable yield in kaolin-treated plants was 21% higher than those measured in control plants; this is possibly related to the 96 and 79% reduction in sunburned fruit and those damaged by insects, respectively, and to the 9% increase in mean fruit weight. Kaolin treatment increased lycopene fruit content by 16%, but did not affect total soluble solids content, fruit dry matter, juice pH, titratable acidity or tomato fruit firmness. The use of kaolin-based particle film technology would be an effective tool to alleviate heat stress and to reduce water stress in tomato production under arid and semi-arid conditions.     

Photosynthetic capacity of field-grown durum wheat under different N availabilities: A comparative study from leaf to canopy

The effect of N availability on photosynthetic capacity, growth parameters and yield was studied in field-grown durum-wheat plants at both the leaf and canopy levels. Two contrasting nitrogen levels (120 and 0 kg ha^?1) were assayed in a randomised block design with nine replicates each. Total biomass was measured at anthesis and yield and its agronomical components at maturity. Photosynthetic measurements were performed 2 weeks after anthesis in two plots of each N treatment. Flag leaves were measured, using a LI-COR 6400 combined with the chlorophyll fluorescence meter, and the whole canopy by measuring CO₂ and H₂O fluxes in an innovative canopy-chamber system. We showed a clear increase in photosynthetic gas exchange and chlorophyll contents with N fertilisation at both canopy and leaf levels. As a consequence the increase in yield as response to N fertilisation seems the result of a larger green leaf area combined with a higher photosynthetic capacity of the leaves attributable to an increase in the maximum carboxylation velocity of Rubisco. Moreover gas-exchange measurements of the flag leaf during grain filling seem to provide a realistic characterisation, not just of the photosynthetic performance of the crop, but also about the impact of N availability on yield. Thus, measurements performed on the flag leaf matched those at the canopy level, with proportional increases in terms of gas exchange and chlorophyll content, providing a fast, cheap and reliable estimation of canopy photosynthesis and the grain yield attained by the crop.     

Prenatal exposure to non-ionizing radiation: effects of WiFi signals on pregnancy outcome, peripheral B-cell compartment and antibody production

During embryogenesis, the development of tissues, organs and systems, including the immune system, is particularly susceptible to the effects of noxious agents. We examined the effects of prenatal (in utero) exposure to WiFi signals on pregnancy outcome and the immune B-cell compartment, including antibody production. Sixteen mated (plug-positive) female mice were assigned to each of the following groups: cage control, sham-exposed and microwave-exposed (WiFi signals at 2.45 GHz, whole body, SAR 4 W/kg, 2 h/day, 14 consecutive days starting 5 days after mating). No effects due to exposure to WiFi signals during pregnancy on mating success, number of newborns/mother and body weight at birth were found. Newborn mice were left to grow until 5 or 26 weeks of age, when immunological analyses were performed. No differences due to exposure were found in spleen cell number, B-cell frequency or antibody serum levels. When challenged in vitro with LPS, B cells from all groups produced comparable amounts of IgM and IgG, and proliferated at a similar level. All these findings were consistently observed in the female and male offspring at both juvenile (5 weeks) and adult (26 weeks) ages. Stress-associated effects as well as age- and/or sex-related differences were observed for several parameters. In conclusion, our results do not show any effect on pregnancy outcome or any early or late effects on B-cell differentiation and function due to prenatal exposure to WiFi signals.     

Effects of Extremely Low-Frequency Electromagnetic Fields on Helicobacter pylori Biofilm

The aim of this work was to investigate the effects of exposure to extremely low-frequency electromagnetic fields (ELF-EMF) both on biofilm formation and on mature biofilm of Helicobacter pylori. Bacterial cultures and 2-day-old biofilm of H. pylori ATCC 43629 were exposed to ELF-EMF (50 Hz frequency–1 mT intensity) for 2 days to assess their effect on the cell adhesion and on the mature biofilm detachment, respectively. All the exposed cultures and the respective sham exposed controls were studied for: the cell viability status, the cell morphological analysis, the biofilm mass measurement, the genotypic profile, and the luxS and amiA gene expression. The ELF-EMF acted on the bacterial population during the biofilm formation displaying significant differences in cell viability, as well as, in morphotypes measured by the prevalence of spiral forms (58.41%) in respect to the controls (33.14%), whereas, on mature biofilm, no significant differences were found when compared to the controls. The measurement of biofilm cell mass was significantly reduced in exposed cultures in both examined experimental conditions. No changes in DNA patterns were recorded, whereas a modulation in amiA gene expression was detected. An exposure to ELF-EMF of H. pylori biofilm induces phenotypic changes on adhering bacteria and decreases the cell adhesion unbalancing the bacterial population therefore reducing the H. pylori capability to protect itself.