Effects of LHRH-analogues on mitogenic signal transduction in cancer cells

The expression of luteinizing hormone-releasing hormone (LHRH) and its receptors has been demonstrated in a number of human malignant tumors, including cancers of the breast, ovary, endometrium and prostate. These findings suggest the presence of an autocrine regulatory system based on LHRH. Recent studies in our laboratory have demonstrated that the function of LHRH produced by ovarian cancer cells is the inhibition of their proliferation. Dose-dependent antiproliferative effects of LHRH-agonists have been observed by several laboratories in cell lines derived from the above cancers. Interestingly, also LHRH-antagonists have marked antiproliferative activity in most of the ovarian, breast and endometrial cancer cell lines tested so far, indicating that the dichotomy of LHRH-agonists/LHRH-antagonists is not valid for the LHRH-system in cancer cells. In addition, our data suggest that the classical LHRH receptor signal transduction mechanisms known from the pituitary (phospholipase-C, protein kinase C, adenylyl cyclase) are not involved in the mediation of LHRH effects in cancer cells. Data obtained by several groups, including ours, rather suggest that LHRH analogs interfere with the signal transduction of growth-factor receptors and related oncogene products associated with tyrosine-kinase activity. The mechanism of action is probably an LHRH-induced activation of a phosphotyrosine phosphatase, counteracting the effects of receptor associated tyrosine kinase. In our hands, LHRH analogs virtually blocked the EGF-induced MAP-kinase activity of ovarian and endometrial cancer cells. The pharmacological exploitation of this mechanism might provide promising new therapies for these cancers.     

On the consistency of individual classification using short scales.

Short tests containing at most 15 items are used in clinical and health psychology, medicine, and psychiatry for making decisions about patients. Because short tests have large measurement error, the authors ask whether they are reliable enough for classifying patients into a treatment and a nontreatment group. For a given certainty level, proportions of correct classifications were computed for varying test length, cut-scores, item scoring, and choices of item parameters. Short tests were found to classify at most 50% of a group consistently. Results were much better for tests containing 20 or 40 items. Small differences were found between dichotomous and polytomous (5 ordered scores) items. It is recommended that short tests for high-stakes decision making be used in combination with other information so as to increase reliability and classification consistency. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Culturing immobilized plant cells for the TUBUL space experiments on the DELTA and 12S Missions

For the TUBUL experiments during the DELTA mission in April 2004 and 12S mission in March/April 2006 on board the Soyuz capsule and the International Space Station we developed a method to culture and chemically fix plant suspension culture cells. The aim of the ten day experiment was to investigate the effect of microgravity on single plant cells. Fully automated experiment cassettes (Plunger Box Units) were developed by Centre for Concepts in Mechatronics (Nuenen, the Netherlands). Tobacco BY- 2 cells were immobilized in a semi- solid agarose matrix that was reinforced by a nylon mesh. This assembly allowed liquid medium refreshment, oxygen supply and chemical fixation, including a post- fixative wash. The method was optimized for post- flight analysis of cell structure,shape and size, cell division, and the microtubule cytoskeleton. The viability of cells in the agarose matrix was similar to cells grown in liquid medium under laboratory conditions, only the stationary growth phase was reached six days later.     

Density and electrical conductivity of molten NaCl-KCl-AlCl3 mixtures

The density and specific electrical conductivity were investigated for molten ternary NaCl-KCl-AlCl₃ mixtures of interest to new industrial processes for electrolytic aluminium deposition. A method for the calculation of density and electrical conductivity of molten ternary mixtures on the basis of experimentally determined values for the binary systems NaCl-AlCl₃, NaCl-KCl and KCl-AlCl₃ has been proposed. The difference between the experimental and calculated density and electrical conductivity values of the ternary system was less than ± 1.5% for the density and ± 5% for the electrical conductivity, respectively.     

Sulfonated poly(aryl ether ketone)s

Several kinds of commercially available polymers of the type poly(oxy-1,4-phenylenecarbonyl-1,4-phenylene) were sulfonated by reaction with mixtures of sulfuric acid and oleum. The ether/carbonyl group ratio of the polymer chain varied from 0.67 to 2.0. With decreasing amounts of ether links in the polymer backbone the sulfonation is hindered and the reaction conditions have to be stronger. To yield the same degree of sulfonation, the sulfuric trioxide concentration of the reaction mixture has to be increased. The achievable degree of substitution is limited, in general, to one sulfonate group per substitutable unit (oxy-phenylene-oxy- or oxy-phenylene-carbonyl unit). In dependence on the polymer structure, polymers with contents of sulfonate groups between 1.2 and 2.0 meq·g^–1 polymer are well soluble in dimethylformamide or N-methylpyrrolidone and it is possible to produce membranes with permselectivities >96% and electrical resistances < 2 Ω cm^–2. The sulfonated polymers were characterized by viscosimetry, sedimentation analysis and ¹³C NMR spectroscopy.     

Development and validation of an HPLC method with fluorescence detection for the determination of fluorescent whitening agents migrating from plastic beverage cups

An HPLC method with fluorescence detection has been developed and validated for the quantification of six fluorescent whitening agents (FWA) in plastic beverage cups after extraction and in food simulants after migration at 70°C for 2 h. The sensitivity of the method was high with LODs ranging from 0.053 to 0.251 μg kg^?1 and LOQs from 0.107 to 0.504 μg kg^?1. Accuracy and precision were highly acceptable, with recoveries greater than 82% and RSDs (%) below 16%. The expanded combined uncertainty was found to be less than 23% for the measurements of all studied FWAs. In extracting the analytes from food contact materials (FCM), accelerated solvent extraction (ASE) and Soxhlet extraction were applied using ethanol as the extraction solvent. The results obtained for FWA in 10 different food plastic cups, made from different polymers, were compared. The ASE technique proved to be faster, more effective and efficient than Soxhlet extraction. Migration tests with official food simulants from Regulation (EU) No 10/2011 showed that the substances studied could potentially migrate using the selected migration conditions. The most pronounced effect was observed in case of simulant D1 (50% w/v ethanol in water). The analytical method proved to be a simple, fast, sensitive and reliable tool for the simultaneous quantification of six of the most used FWAs in both FCM extracts and food simulants after migration experiments.     

Global, Local, and Graphical Person-Fit Analysis Using Person-Response Functions.

Person-fit statistics test whether the likelihood of a respondent's complete vector of item scores on a test is low given the hypothesized item response theory model. This binary information may be insufficient for diagnosing the cause of a misfitting item-score vector. The authors propose a comprehensive methodology for person-fit analysis in the context of nonparametric item response theory. The methodology (a) includes H. Van der Flier's (1982) global person-fit statistic U3 to make the binary decision about fit or misfit of a person's item-score vector, (b) uses kernel smoothing (J. O. Ramsay, 1991) to estimate the person-response function for the misfitting item-score vectors, and (c) evaluates unexpected trends in the person-response function using a new local person-fit statistic (W. H. M. Emons, 2003). An empirical data example shows how to use the methodology for practical person-fit analysis. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Feasibility study for producing a carrot/potato matrix reference material for 11 selected pesticides at EU MRL level: Material processing, homogeneity and stability assessment

The feasibility for producing a matrix reference material for selected pesticides in a carrot/potato matrix was investigated. A commercially available baby food (carrot/potato-based mash) was spiked with 11 pesticides at the respective EU maximum residue limits (MRLs), and further processed by either freezing or freeze-drying. Batches of some 150 units were produced per material type. First, the materials were assessed for the relative amount of pesticide recovered after processing (ratio of pesticide concentration in the processed material to the initially spiked pesticide concentration). In addition, the materials’ homogeneity (bottle-to-bottle variation), and the short-term (1 month) and mid-term (5 months) stability at different temperatures were assessed. For this, an in-house validated GC-EI-MS method operated in the SIM mode with a sample preparation procedure based on the QuEChERS (“quick, easy, cheap, effective, rugged, and safe”) principle was applied. Measurements on the frozen material provided the most promising results (smallest analyte losses during production), and also freeze-drying proved to be a suitable alternative processing technique for most of the investigated pesticides. Both the frozen and the freeze-dried material showed to be sufficiently homogeneous for the intended use, and storage at −20 °C for 5 months did not reveal any detectable material degradation. The results constitute an important step towards the development of a pesticide matrix reference material.     

Rates of exocytosis and endocytosis in Arabidopsis root hairs and pollen tubes

Exocytosis and endocytosis are pivotal in many biological processes, but remain difficult to quantify. Here we combine a new algorithm for estimating vesicle size with a detailed morphological analysis of tip-growing cells, in which exocytosis is highly localized and therefore more readily quantified. Cell preservation was rendered as life-like as possible by rapid freezing. This allowed us to produce the first estimates of exocytosis rates in the root hairs and pollen tubes of the model plant Arabidopsis. To quantify exocytosis and endocytosis rates during cell growth, we measured the diameter of vesicles located in the tips of Arabidopsis root hairs and pollen tubes and the widths of cell walls and the cell lumen in longitudinal thin transmission electron microscopic sections. In addition, we measured growth velocities of Arabidopsis root hairs and pollen tubes, using video microscopy. The number of exocytotic vesicles required for cell wall expansion, and the amount of excess membrane inserted into the plasma membrane to be internalized, were estimated from the values that were obtained. The amount of excess membrane that is inserted into the plasma membrane during cell growth was estimated as 86.7% in root hairs and 79% in pollen tubes. This membrane has to be recycled by endocytosis. From counting of the total number of vesicles that is present in thin EM sections through the pollen tube tip, we estimated the average number of vesicles that is present in the tip of pollen tubes. By calculating the total amount of membrane and cell wall material that is required for continued cell growth, assuming that all vesicles are exocytotic, we estimated that pollen tubes continue to grow for 33 s when delivery of vesicles to the tip is inhibited. We arrested vesicle delivery to the tip by application of cytochalasin D. After cytochalasin D application, pollen tubes continued to grow for 30-40 s, which is in the same range as the estimated value of 33 s and shows that in this time frame, the availability of exocytotic vesicles is not a limiting factor.