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.     

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     

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.     

Screening of supports for the immobilization of pectinmethylesterase from acerola (Malpighia glabra L)

A partially purified extract of pectinmethylesterase (PME) from acerola fruit was immobilized on various supports: glass, celite, chrysotile, agarose, concanavalin A Sepharose 4B, egg shell, polyacrylamide and gelatin. In addition, reticulation with glutaraldehyde was assessed, as well as the use of gelatin in the presence of celite, glass and silica. The highest immobilization yields were obtained when the pectinmethylesterase was immobilized in concanavalin A Sepharose 4B (81.7%) and in gelatin‐water (78.0%). Copyright © 2004 Society of Chemical Industry     

Effects of temperature, ammonium and glucose concentrations on yeast growth in a model wine system

In enology, alcoholic fermentation is a complex process involving several mechanisms. Slow and incomplete alcoholic fermentation is a chronic problem for the wine industry and factors leading to sluggish and stuck fermentations have been extensively studied and reviewed. The most studied cause of sluggish and stuck fermentation is the nitrogen content limitation. Nevertheless, other factors, such as temperature of fermentation and sugar concentration can affect the growth of yeasts. In this study we modelled the yeast growth‐cycle in wine model system as a function of temperature, sugar and ammonium concentrations; the individual effects and the interaction of these factors were analysed by means of a quadratic response surface methodology. Cell concentrations and weight loss were monitored in the whole wine fermentation process. The results of central composite design show that lower is the availability of nitrogen, higher is the cell growth rate; moreover, initial nitrogen concentration also influences survival time of Saccharomyces cerevisiae.     

Thermo-oxidative degradation of some polymer couples containing HNBR

Summary In this paper thermal behavior of some HNBR blends with CPE, PVC, CR and NBR at three temperatures, 150, 170 and 190°C is presented. Changes in characteristic IR bands, gel fraction, and chlorine content were assessed. Some mechanistic differences between the studied systems, regarding the involvement the depletion of chlorine and modification in gel content, were pointed out. Cyclization of nitrile structure was also observed by means of 2240 cm⁻¹ band ascribed to >C=NH. Kinetic considerations are presented. Received: 3 June 2002/Revised version: 14 November 2002/ Accepted 20 November 2002 Correspondence to Traian Zaharescu     

Effects of different cooking methods on the formation of food mutagens in meat

Hamburgers and chicken fillets were cooked in convection ovens, deep‐fried or contact fried and analysed for mutagenic activity using the Ames test. For the three different convection ovens, the cooking parameters studied included the presence of steam, air velocity, air temperature and holding time. For deep‐frying and contact frying, the cooking parameters included cooking temperature and cooking time. In cooked hamburgers, mutagenic activity was only detected in those that had been deep‐fried. In chicken fillets, mutagenic activity was detected in samples prepared with all cooking methods, being highest in the deep‐fried samples. Factorial analysis indicated that heat transfer was the most important factor affecting mutagenic activity. High temperature and high air velocity in the convection ovens enhanced mutagenic activity. The presence of steam reduced the mutagenic activity, except when high temperature was used in combination with high air velocity. In chicken fillets, high mutagenic activity correlated to high weight loss during cooking. Pan‐fried chicken fillets were analysed using high performance liquid chromatography (HPLC) and the mutagenic/carcinogenic heterocyclic amines 2‐amino‐3,8‐dimethylimidazo[4,5‐f ]‐quinoxaline (MeIQx), 2‐amino‐1‐methyl‐6‐phenylimidazo‐[4,5‐b]‐pyridine (PhIP) and the co‐mutagenic heterocyclic amine Norharman (9H‐pyrido[3,4‐b]‐indole) were identified. The HPLC fractions were tested for mutagenic activity and, apart from the mutagenic fractions corresponding to MeIQx and PhIP, several mutagenic fractions were detected that did not correspond to known heterocyclic amines.