Quantitative methods for scoring cell migration and invasion in filter-based assays

The ability of cells to traverse pores in a biocompatible filter provides means for examining cell chemoattraction. Filter-based assays also permit rapid, quantitative assessment of the in vitro migratory and invasive potential of tumor cells. Scoring migration has relied on visual counting of stained cells which appear on the underside of the filter and determining a true percentage score involves arduous counting of cells on both filter surfaces. Visual counting of random fields may be unreliable, and counting all fields is laborious. In the present study we developed and compared two alternative methods for scoring cell numbers in filter-based assays, a colorimetric assay of toluidine blue binding, and a radioassay of cells prelabeled with [3H]thymidine. Each method was evaluated for sensitivity, variability, ease of use and efficiency, and suitability for use in assays of cell migration and invasion. The radiolabeling method proved to be sensitive and reliable and was the most efficient technique. Although less sensitive and specific, the colorimetric dye method offered a rapid and reliable, nonradioactive alternative with the distinct advantage of preserving intact cultures for follow-up visual assessments. We conclude that colorimetric and radiolabel scoring of filter-based assays are reliable and efficient semiautomated methods which provide means to obtain more complete assessments of cell migration and invasion.     

Medium and substitution pattern effects on the action of hydroxyflavones as photoprotectors against singlet molecular oxygen-mediated photooxidation of fats

The ability of mono-, di-, tri-and tetra-hydroxyflavones (7-and 3-hy-droxyflavones, 7, 8-dihydroxyflavone, baicalein and fisetin) to act as photoprotectors against singlet molecular oxygen [O²(Δ_g)]-initiated photooxidation of fats has been established by a kinetic study. The overall quenching rate constants for a series of five hydroxyflavones perfectly parallel their respective behaviour as inhibitors of the sensitized photooxidation of linoleic acid. The best antioxidative effect was exerted by 7-hydroxyflavone which does not chemically react with O²(¹Δ_g). Nevertheless for the remaining flavonoids of the series, the physical deactivation of O²(¹Δ_g) largely prevails over the chemical process. As for the cases of phenols and related hydroxy-aromatic derivatives, the ionization of the -OH group in the flavones, dramatically accelerates the rate of photooxidation. Under these conditions, the 7-hydroxyflavone also reacts effectively with O²(¹Δ_g). Given that flavonoids are natural oil components, this medium effect should be taken into account during the oil-refining process, in order to avoid the flavonoid photodestruction.     

Molecular dynamics simulations of polyphosphazenes: poly[bis(chloro)phosphazene][NPCl2] n

Suitable parameter sets for the CHARMm force field were derived for the structural units in polychlorophosphazene [P=N, P N, P Cl] using the Dinur Hagler energy second derivative procedure based on quantum mechanical SCI calculations using the 6–31G^* basis set. To validate the reliability of the parameter set, structural results obtained with CHARMm for the adopted model compounds (OP₂NCl₅ and OP₃N₂Cl₅) were compared with those derived fromab initio quantum mechanics using the 6–31G^* basis set. Application of molecular dynamics (MD) simulations in combinatioin with the available X-ray diffraction data provided structural and conformational information on the polymer. The calculation made using the periodic boundary conditions (PBC) agree well with the polychlorophosphazene ordered in a monoclinic unit cell (a=5.98,b=12.99,c=4.92 A; β=111.7). This model was stabilized mainly by the image atoms contribution to the electrostatic energy term and had aquasi-planar conformation of the backbone chain (glide symmetry). The MD calculations also provided evidence that the difference between single and double PN bonds is less marked than that measured experimentally. This result is, however, in agreement with more recent and accurate X-ray studies on poly(methylphosphazene). Validation of the polymer model provided a complete picture, otherwise experimentally inaccessible, of the internal fluctuations of the polymeric chains.     

Enhancement mode InP MISFET's with sulfide passivation andphoto-CVD grown P3N5 gate insulators

High performance enhancement mode InP MISFET's have been successfully fabricated by using the sulfide passivation for lower interface states and with photo-CVD grown P₃N₅ film used as gate insulator. The MISFET's thus fabricated exhibited exhibited pinch-off behavior with essentially no hysteresis. Furthermore the device showed a superior stability of drain current. Specifically under the gate bias of 2 V for 10⁴ seconds the room temperature drain current was shown to reduce from the initial value merely by 2.9% at the drain voltage of 4 V. The effective electron mobility and extrinsic transconductance are found to be about 2300 cm ²/V·s and 2.7 mS/mm, respectively. The capacitance-voltage characteristics of the sulfide passivated InP MIS diodes show little hysteresis and the minimum density of interface trap states as low as 2.6×10¹⁴/cm² eV has been attained     

Bulk modification of styrene–butadiene–styrene triblock copolymer with maleic anhydride

The bulk modification of SBS rubber with maleic anhydride in a mixing chamber of a Haake rheomixer was studied. The effect of temperature, maleic anhydride, and benzoyl peroxide concentrations on the grafting efficiency was evaluated. High grafting efficiency was achieved when the ratio of peroxide and maleic anhydride concentration was high. On the other hand, on this condition high insoluble fraction was generated. The addition of a diamine, 4,4′‐diaminediphenylmethane to the reaction mixture minimizes the amount of insoluble polymer. However, the grafted MAH content also decreases. The graft copolymer was characterized by infrared spectroscopy and the grafting extension was determined by titration. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 2953–2960, 2002; DOI 10.1002/app.10355     

Pressure effects on flow and fracture of Be-Al alloys

The flow and fracture behavior of Be-Al alloys were determined in tension with different levels of superimposed pressure. The Be-Al alloys were prepared by Brush Wellman, Inc. (Cleveland, OH) from prealloyed powders processed to either a hot isostatically pressed (“hipped”) or cold isostatically pressed and extruded condition. Significant effects of pressure on both the flow and ductility have been observed at room temperature, with implications on the formability of these materials. The effects of changes in processing conditions and stress state on the flow and fracture behavior are summarized in addition to both optical and scanning electron microscopy (SEM) examination of the fracture surfaces. Separate other studies on the alloy constituents (e.g., Al and Be) are also reported. The results are also compared to previous works on monolithic materials and composites tested with high pressure.     

Spatial learning deficits in mice with a targeted glucocorticoid receptor gene disruption

Previous studies in rats using the Morris water maze suggested that the processing of spatial information is modulated by corticosteroid hormones through mineralocorticoid and glucocorticoid receptors in the hippocampus. Mineralocorticoid receptors appear to be involved in the modulation of explorative behaviour, while additional activation of glucocorticoid receptors facilitates the storage of information. In the present study we used the water maze task to examine spatial learning and memory in mice homozygous and heterozygous for a targeted disruption of the glucocorticoid receptor gene. Compared with wild-type controls, homozygous and heterozygous mice were impaired in the processing of spatial but not visual information. Homozygous mutants performed variably during training, without specific platform-directed search strategies. The spatial learning disability was partly compensated for by increased motor activity. The deficits were indicative of a dysfunction of glucocorticoid receptors as well as of mineralocorticoid receptors. Although the heterozygous mice performed similarly to wild-type mice with respect to latency to find the platform, their strategy was more similar to that of the homozygous mice. Glucocorticoid receptor-related long-term spatial memory was impaired. The increased behavioural reactivity of the heterozygous mice in the open field points to a more prominent mineralocorticoid receptor-mediated function. The findings indicate that (i) the glucocorticoid receptor is of critical importance for the control of spatial behavioural functions, and (ii) mineralocorticoid receptor-mediated effects on this behaviour require interaction with functional glucocorticoid receptors. Until the development of site-specific, inducible glucocorticoid receptor mutants, glucocorticoid receptor-knockout mice present the only animal model for the study of corticosteroid-mediated effects in the complete absence of a functional receptor.     

Ionomer composites based on sepiolite/hydrogenated poly(styrene butadiene) block copolymer systems

Ionomeric composites based on sepiolite and hydrogenated poly(styrene butadiene) block copolymer were obtained and characterized from a microstructural and electrical point of view. Before blending, because of the high silanol group concentration in the sepiolite, the latter could be organophilized with suitable coupling agents. The resulting materials were easily processed into thin films or membranes 0.2–0.4 mm thick, their conductivity in some cases approaching 10^?1 S/cm. Their suitability for film formation and good electrical properties indicate potential applications as electrolytes in polymer fuel cells. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 3512–3519, 2002     

Primary lymphoma of the breast. Three cases of non-Hodgkin lymphoma

Primary and sole breast lymphoma is a very rare disease. With the review of our series of 616 cases operated on for breast cancer, only 3 cases (0.48%) of primary breast non-Hodgkin lymphoma (LNH) have been observed. The authors outline the problems concerning diagnosis and therapy of this rare disease: pathological and immunohistochemical aspects are discussed. They stress the importance of staging in order to plan a correct multidisciplinary approach.