Rapid mutation detection in complex genes by heteroduplex analysis with capillary array electrophoresis

Mutational analysis of large multiexon genes without prevalent mutations is a laborious undertaking that requires the use of a high-throughput scanning technique. The Human Genome Project has enabled the development of powerful techniques for mutation detection in large multiexon genes. We have transferred heteroduplex analysis (HA) by conformation-sensitive gel electrophoresis of the two major breast cancer (BC) predisposing genes, BRCA1 and BRCA2, to a multicapillary DNA sequencer in order to increase the throughput of this technique. This new method that we have called heteroduplex analysis by capillary array electrophoresis (HA-CAE) is based on the use of multiplex-polymerase chain reaction (PCR), different fluorescent labels and HA in a 16-capillary DNA sequencer. To date, a total of 114 different DNA sequence variants (19 insertions/deletions and 95 single-nucleotide substitutions - SNS) of BRCA1 and BRCA2 from 431 unrelated BC families have been successfully detected by HA-CAE. In addition, we have optimized the multiplex-PCR conditions for the colorectal cancer genes MLH1 and MSH2 in order to analyze them by HA-CAE. Both genes have been amplified in 13 multiplex groups, which contain the 35 exons, and their corresponding flanking intronic sequences. MLH1 and MSH2 have been analyzed in nine hereditary nonpolyposis colorectal cancer patients, and we have found six different DNA changes: one complex deletion/insertion mutation in MLH1 exon 19 and another five SNS. Only the complex mutation and one SNS may be classified as cancer-prone mutations. Our experience has revealed that HA-CAE is a simple, fast, reproducible and sensitive method to scan the sequences of complex genes.     

Silica-Zirconia Coatings Produced by Dipping and EPD from Colloidal Sol–Gel Suspensions

SiO₂-ZrO₂ sols have been prepared via acid catalysis using a commercial colloidal suspension of zirconia and two silica alkoxides; tetraethoxysilane (TEOS) and methyltriethoxysilane (MTES). Suspensions with 10, 15 and 25-mol% of ZrO₂ were prepared. The stability of the suspensions was followed by rheological measurements showing that the amount of water incorporated with the colloidal suspension is the factor that limits the maximum ZrO₂ content. Coatings have been prepared by dipping using the suspensions up to 25-mol% ZrO₂ onto glass-slides at different withdrawal rates. EPD process has been used to prepare coatings onto stainless steel AISI 304 using the suspension with 25-mol% ZrO₂ at different pHs. The parameters associated with the EPD process (current density, electric field, potential and deposition time) have been evaluated. The critical thickness for a ZrO₂ addition of 25-mol% was 0.8 μm and it increased for diminishing ZrO₂ content.     

Assembly of a Thin-Falling-Film Exchanger for Laboratory Demonstrations: Calculation of the Individual Heat-Transfer Coefficient

Thin-film exchangers have a large heat-transfer area per unit liquid volume that make them very efficient for industrial use. Unfortunately, they are usually less applied for education purposes. In this work, a simple experimental device with a fluid flowing as a falling film has been assembled. In addition, an experimental guide allows students to determine the individual heat-transfer coefficient in the exchanger.The experimental setup consists of two pipes bundled as a shell-and-tube heat exchanger. The cold liquid flows in the form of a thin film that runs down inside the inner tube; the hot liquid flows down concurrently in the external one.In order to test the operation of the device, experimental results are given for a simple test with water. Heat-transfer coefficients are calculated and a theoretical correlation obtained from the literature is compared with the experimental results. In addition, a fit of the data allows for the formulation of a new empirical equation for the transient regime.     

Does the D(-)/D(+) production asymmetry decrease at large x(F)?

We have applied the meson cloud model to calculate the asymmetries in D and D(s) meson production in high-energy Sigma(-)-nucleus and pi(-)-nucleus collisions. We find a good agreement with recent data. Our results suggest that the asymmetries may decrease at large x(F).     

String effects on Fermi–Dirac correlation measurements

We investigate some recent measurements of Fermi–Dirac correlations by the LEP collaborations indicating surprisingly small source radii for the production of baryons in e⁺e^- annihilation at the Z⁰ peak. In hadronization models there is besides the Fermi–Dirac correlation effect also a strong dynamical (anti-) correlation. We demonstrate that the extraction of the pure FD effect is highly dependent on a realistic Monte Carlo event generator, both for separation of those dynamical correlations that are not related to Fermi–Dirac statistics, and for corrections of the data and background subtractions. Although the model can be tuned to well reproduce single particle distributions, there are large model uncertainties when it comes to correlations between identical baryons. We therefore, unfortunately, have to conclude that it is at present not possible to draw any firm conclusion about the source radii relevant for baryon production at LEP. PACS 13.60.Rj; 13.66.Bc     

Biological Activities of Violacein, a New Antitumoral Indole Derivative, in an Inclusion Complex with β-Cyclodextrin

Violacein is a poorly water-soluble antitumoraland antibacterial drug. The solubility can be enhanced by complexation with-cyclodextrin. The inclusion complex was prepared by the co-precipitation method in molarratios of 1 : 1 and 1 : 2 of violacein/-cyclodextrin, respectively. The acutetoxicity (E. coli strain) of violacein did not changeup to 400 M, either in thepresence or absence of cyclodextrin. Cytotoxicity (V-79 cellculture) through DNA and MTT assays was significantlydecreased in the presence of the 1 : 2 molar ratio complex.Studies on erythrocyte lipid peroxidation by the thiobarbituricacid (TBA) methodshowed that violacein and violacein/-CD (1 : 2) at100 M cause 50% and 80% inhibition, respectively. At 500 M theviolacein/-CD complexinhibited lipid peroxidation completely; however, withfree violacein only 65% inhibition was reached at that concentration.     

A transformation from Hausdorff to Stieltjes moment sequences

We introduce a non-linear injective transformation τ from the set of non-vanishing normalized Hausdorff moment sequences to the set of normalized Stieltjes moment sequences by the formulaT[(a _n ) _n=1 ^∞ ] _n = 1/a₁ ...a _n . Special cases of this transformation have appeared in various papers on exponential functionals of Lévy processes, partly motivated by mathematical finance. We give several examples of moment sequences arising from the transformation and provide the corresponding measures, some of which are related toq-series.     

Synthesis of enantiopure 3-azabicyclo[3.3.0]octen-7-one derivatives via intramolecular Pauson-Khand cycloaddition reaction using tri-O-acetyl-D-glucal as starting material

Some N-substituted-3-azabicyclo[3.3.0]octen-7-one derivatives have been synthesized in an enantiomerically pure form starting from tri-O-acetyl-D-glucal via intramolecular Pauson-Khand (IPK) cycloaddition.