Laser flow measurements of scattering and fluorescence from cell nuclei in the presence of increasing Mg++ concentrations.

The effects of Mg++ on the spatial organization of nuclei from rat hepatocytes are analyzed in the range 0-60 mM, in the presence of suitable concentrations of KCl to reproduce physiological conditions. It is shown that the scatter-signal distribution measured by means of a flow microfluorimeter is greatly affected by this range of Mg concentrations. By coupling this result to phase-contrast-automated image analysis, it is possible to identify a shrinking process induced by Mg++ in the range 0-2.5 mM, which reaches a plateau in the range 5-20 mM and is followed by a swelling process in the range 30-60 mM. The same Mg ranges are shown to affect the intercalation of the fluorochrome acridine orange into chromatin, suggesting that the shrinking-swelling phenomenon has also a molecular correspondence at the genome level. Possible implications in terms of the influence of Mg++ on the organization of chromatin inside intact cells are briefly discussed.     

Amyloid beta(1-42) and its beta(25-35) fragment induce activation and membrane translocation of cytosolic phospholipase A2 in bovine retina capillary pericytes

We investigated changes in cytosolic phospholipase A(2) (cPLA(2)) and calcium-independent PLA(2) (iPLA(2)) activities in bovine retina capillary pericytes after stimulation with 50 microM amyloid-beta (Abeta) (1-42) and its (25-35) fragment, over 24 h (mild, sublethal model of cell damage). In the presence of Abeta peptides, we found that cPLA(2) activity was increased and translocated from the cytosolic fraction to the membrane system, particularly in the nuclear region. Reversed-sequence Abeta(35-25) peptide did not stimulate or induce cPLA(2) translocation. Exposure to both Abeta peptides had no significant effect on cPLA(2) protein content as tested by Western immunoblot analysis. The addition of Abetas to quiescent pericytes was followed by phosphorylation of cPLA(2) and arachidonic acid release. Treatment with inhibitors (AACOCF(3), staurosporine and cycloheximide) resulted in a sharp decrease in basal and stimulated cPLA(2) activity. Inactivating effects of bromoenol lactone (BEL), inhibitor of iPLA(2), demonstrated that the stimulation of total PLA(2) activity by Abetas was mediated by both PLA(2) enzymes. Taken together with our previous observations that both Abeta peptides may induce hydrolysis of phosphatidylcholine, the present results provide evidence that this process is cooperatively mediated by cPLA(2) activation/translocation and iPLA(2) activation. The effect is very likely triggered by a mild prooxidant mechanism which was not able to divert the cell to degeneration. The data confirm the hypothesis that pericytes could be a target of potential vascular damage and reactivity during processes involving amyloid accumulation.     

HFE and transferrin directly compete for transferrin receptor in solution and at the cell surface

Transferrin receptor (TfR) is a dimeric cell surface protein that binds both the serum iron transport protein transferrin (Fe-Tf) and HFE, the protein mutated in patients with the iron overload disorder hereditary hemochromatosis. HFE and Fe-Tf can bind simultaneously to TfR to form a ternary complex, but HFE binding to TfR lowers the apparent affinity of the Fe-Tf/TfR interaction. This apparent affinity reduction could result from direct competition between HFE and Fe-Tf for their overlapping binding sites on each TfR polypeptide chain, from negative cooperativity, or from a combination of both. To explore the mechanism of the affinity reduction, we constructed a heterodimeric TfR that contains mutations such that one TfR chain binds only HFE and the other binds only Fe-Tf. Binding studies using a heterodimeric form of soluble TfR demonstrate that TfR does not exhibit cooperativity in heterotropic ligand binding, suggesting that some or all of the effects of HFE on iron homeostasis result from competition with Fe-Tf for TfR binding. Experiments using transfected cell lines demonstrate a physiological role for this competition in altering HFE trafficking patterns.     

Crk-associated substrate tyrosine phosphorylation sites are critical for invasion and metastasis of SRC-transformed cells

Crk-associated substrate (CAS, p130Cas) is a major tyrosine phosphorylated protein in cells transformed by v-crk and v-src oncogenes. We recently reported that reexpression of CAS in CAS-deficient mouse embryo fibroblasts transformed by oncogenic Src promoted an invasive phenotype associated with enhanced cell migration through Matrigel, organization of actin into large podosome ring and belt structures, activation of matrix metalloproteinase-2, and elevated tyrosine phosphorylation of the focal adhesion proteins FAK and paxillin. We have now extended these studies to examine the mechanism by which CAS achieves these changes and to evaluate the potential role for CAS in promoting in vivo tumor growth and metastasis. Whereas the presence or absence of CAS did not alter the primary growth of subcutaneous-injected Src-transformed mouse embryo fibroblasts, CAS expression was required to promote lung metastasis following removal of the primary tumor. The substrate domain YxxP tyrosines, the major sites of CAS phosphorylation by Src that mediate interactions with Crk, were found to be critical for promoting both invasive and metastatic properties of the cells. The ability of CAS to promote Matrigel invasion, formation of large podosome structures, and tyrosine phosphorylation of Src substrates, including FAK, paxillin, and cortactin, was also strictly dependent on the YxxP tyrosines. In contrast, matrix metalloproteinase-2 activation was most dependent on the CAS SH3 domain, whereas the substrate domain YxxP sites also contributed to this property. Thus multiple CAS-mediated signaling events are implicated in promoting invasive and metastatic properties of Src-transformed cells.     

Liquid Norbornadiene Photoswitches for Solar Energy Storage

Due to high global energy demands, there is a great need for development of technologies for exploiting and storing solar energy. Closed cycle systems for storage of solar energy have been suggested, based on absorption of photons in photoresponsive molecules, followed by on‐demand release of thermal energy. These materials are called solar thermal fuels (STFs) or molecular solar thermal (MOST) energy storage systems. To achieve high energy densities, ideal MOST systems are required either in solid or liquid forms. In the case of the latter, neat high performing liquid materials have not been demonstrated to date. Here is presented a set of neat liquid norbornadiene derivatives for MOST applications and their characterization in toluene solutions and neat samples. Their synthesis is in most cases based on solvent‐free Diels‐Alder reactions, which easily and efficiently afford a range of compounds. The shear viscosity of the obtained molecules is close to that of colza oil, and they can absorb up to 10% of the solar spectrum with a measured energy storage density of up to 577 kJ/kg corresponding to 152 kJ mol^–1 (calculated 100 kJ mol^–1). These findings pave the way towards implementation of liquid norbornadienes in closed cycle energy storage technologies.     

Age and growth of the nototheniid fishTrematomus bernacchii Boulenger from Terra Nova Bay,Antarctica

Age and growth of the nototheniid fishTrematomus bernacchii Boulenger 1902 were estimated by reading the sagittal otoliths of 457 adult specimens caught off Terra Nova Bay (Ross Sea) in the austral summer 1990–1991. Annuli in ground and polished otoliths were examined using a dissecting microscope under reflected light. The Von Bertalanffy growth equation was L_t=273.5 [1 − e^{−0.109(t+2.10)}] for males (n=122) and L_t=422.2 [1 − e^{−0.055(t+1.92)}] for females (n=211) where L is total length in millimetres. Maximum estimated age was 21 years for females and 16 years for males. This is in agreement with the hypothesis that considers slow growth and old age as a typical feature of Antarctic fishes.     

Age determination and growth of turbot and brill in the Adriatic Sea: reversal of the seasonal pattern of otolith zone formation

The growth of two commercially important flatfish, turbot ( Psetta maxima ) (L.) and brill ( Scophthalmus rhombus ) (L.), was investigated in the Adriatic using whole otoliths (sagittae) and stained otolith sections. At variance with the pattern usually observed in temperate seas, the opaque zone was found to be laid down in autumn and winter, and the translucent zone in spring and summer. Growth rates differed according to sex, with the females attaining greater body lengths. The von Bertalanffy growth parameters were: L_∞=66.2 cm, K=0.31 years^–1, and t₀=–0.14 years for turbot males, L_∞=81.5 cm, K=0.21 years^–1, and t₀=–0.48 years for turbot females; L_∞=40.2 cm, K=0.49 years^–1, and t₀=–1.03 years for brill males; L_∞=50.1 cm, K=0.27 years^–1, and t₀=–1.75 years for brill females. Growth rates and maximum age recorded for turbot were comparable to those reported in the North Sea.     

NCAM1 is the Target of miRNA-572: Validation in the Human Oligodendroglial Cell Line

The neural cell adhesion molecule 1 (NCAM1) is a fundamental protein in cell–cell interaction and in cellular developmental processes, and its dysregulation is involved in a number of diseases including multiple sclerosis. Studies in rats suggest that the modulation of NCAM1 expression is regulated by miRNA-572, but no data are available confirming such interaction in the human system. We analyzed whether this is the case using a human oligodendroglial cell line (MO3.13). MO3.13 cells were transfected with miRNA-572 mimic and inhibitor separately; NCAM1 mRNA and protein expression levels were analyzed at different time points after transfection. Results indicated that NCAM1 expression is increased after transfection with miRNA-572 inhibitor, whereas it is decreased after transfection with the mimic (p < 0.005). The interaction between NCAM1 and miRNA-572 was subsequently confirmed in a Vero cell line that does not express NCAM1, by luciferase assay after transfection with NCAM1. These results confirm that miRNA-572 regulates NCAM1 and for the first time demonstrate that this interaction regulates NCAM1 expression in human cells. Data herein also support the hypothesis that miRNA-572 is involved in diseases associated with NCAM1 deregulation, suggesting its possible use as a biomarker in these diseases.     

Influence of climate,sea-level changes and tectonics on ichnoassemblages distribution in a carbonate-dominated,deep-marine environment (Upper Paleocene,Zumaya section)

A detailed bed-by-bed ichnological analysis of the Paleocene of the Zumaya section (northern Spain) revealed the presence of eight different ichnoassemblages. Four different sets of beds were determined using both ichnological and sedimentological features. Their distribution throughout the section can be compared to the sedimentary trends based on geochemical and sedimentological data, in order to better understand the ecological significance of the ichnoassemblages and to determine the most important parameters that drove the environmental evolution of the area. Significantly, there is a marked relationship between the distribution of the ichnoassemblages and the recorded variations in the rate of sedimentation in both the background (hemipelagic limestones) and the episodic sediments (calcareous and siliciclastic turbidites). However, some ichnoassemblages (e.g. ichnoassemblage 4a: Planolites beverleyensis and ichnoassemblage 2b: ?Palaeophycus tubularis) while clearly lithofacies-related also appear to be controlled by other parameters, such as the amount of benthic food introduced into the basin from shallow-water areas.