Evaluation of a dual-color flow cytometry immunophenotyping panel in a multicenter quality assurance program

A basic immunophenotyping panel that employed dual-color combinations of fluorescein isothiocyanate (FITC) and phycoerythrin (PE) conjugated monoclonal antibodies (mAb; FITC-CD45/PE-CD14, FITC-IgG1/PE-IgG2, FITC-CD3/PE-CD8, FITC-CD3/PE-CD4, FITC-CD3/PE-CD16 + PE-CD56, and PE-CD19) was utilized in a quality assurance program to determine whether the 4 laboratories participating in a multicenter AIDS study obtained similar lymphocyte subset percentage values for T cells, B cells, NK cells, and CD4+ and CD8+ T cells. Over a 1 1/2 year period, 78 shared peripheral blood specimens were prepared and analyzed in each laboratory. The CD45bright CD14- percentage for each specimen was used to correct that individual's lymphocyte subset values. Interlaboratory coefficients of variation (CV) for the human immunodeficiency virus type I (HIV) seronegative (n = 38) and HIV-seropositive (n = 40) specimens using this panel were < 3% for total T cells; < 5% for CD4+ T cells and CD8+ T cells; < or = 17% for B and NK cells; and < 8% for CD4T/CD8T ratios. The 6-tube basic immunophenotyping panel has several notable features: a) for clinical studies, it permits comprehensive evaluation of an individual's major lymphocyte subsets, i.e., T, B, NK, and CD4+ and CD8+ T cells; b) for interlaboratory proficiency testing programs, it allows the detection of differences among laboratories in measurements of several functionally distinct cell populations; and c) for within-sample quality assurance, it provides several quality control checks, including the lymphosum, i.e., the sum of an individual's corrected T+B+NK values, a sum that was generally 100 +/- 5% on the HIV-seronegative specimens analyzed in this study.     

Modulation of the allergic immune response in BALB/c mice by subcutaneous injection of high doses of the dominant T cell epitope from the major birch pollen allergen Bet v 1

Several in vitro and in vivo studies indicate that application of high doses of dominant T cell epitopes can induce a state of antigen-specific non-responsiveness (anergy). In the present study, we developed a murine model of an allergic immune response to Bet v 1, the major birch pollen allergen. Mice were sensitized by injection of rBet v 1 and the allergic state was proven by the presence of allergen-specific IgE and positive immediate-type skin tests to Bet v 1. In epitope mapping experiments, an immunodominant T cell epitope of Bet v 1 in BALB/c mice was identified by the use of overlapping peptides. This peptide (BV 139) was subsequently employed for treatment. Two tolerization protocols were used: in one approach, the peptide was administered to naive mice before immunization (group BV139-S), in the second, already sensitized mice were treated (S-BV139). The results demonstrated that administering high doses of the dominant T cell epitope of Bet v 1 profoundly diminished T cell proliferation to the peptide in the BV139-S group, and to the peptide as well as to the whole protein in the S-BV139 group. Skin test reactivity to Bet v 1 was reduced in the BV139-S group. However, no differences in terms of specific antibody production between treated and untreated mice could be observed. This study provides evidence that administration of dominant T cell epitopes can down-regulate the allergen-specific T cell response. Proceeding on the assumption that the T lymphocyte response to allergens is crucial for the induction and maintenance of the allergic disease, a modulation of the immune response to allergens by treatment with T cell epitope peptides could represent a promising concept for immunotherapy in the future.     

Formal program construction by transformations-computer-aided,intuition-guided programming

Formal program construction by transformations is a method of software development in which a program is derived from a formal problem specification by manageable, controlled transformation steps which guarantee that the final product meets the initial specification. This methodology has been investigated in the Munich project CIP (computer-aided intuition-guided programming). The research includes the design of a wide-spectrum language specifically tailored to the needs of transformational programming, the construction of a transformation system to support the methodology, and the study of transformation rules and other methodological issues. Particular emphasis has been laid on developing a sound theoretical basis for the overall approach     

Activation of action rules in action observation.

Visually perceiving an action may activate corresponding motor programs. This automatic motor activation can occur both for higher level (i.e., the goal of an action) and for lower level (i.e., the specific effector with which it is executed) aspects of an action. The authors used a tool-use action paradigm to experimentally dissociate priming effects for observing the target, the movement, or the target-to-movement mapping of a tool-use action. In 3 experiments, participants took turns in acting, observing the tool-use action of another person in trial n-1, and executing an action in trial n. Trial transitions from n-1 to n were manipulated in 4 conditions with (a) mapping repeated and movement and target changed, (b) target repeated and movement and mapping changed, (c) movement repeated and target and mapping changed, or (d) all components repeated. Results indicate priming effects for repeating the target-to-movement mapping (i.e., the action rule) of a tool-use action and suggest that a rather abstract action schema is activated during action observation. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

IR spectroscopic study of Pt/Kl zeolites using adsorption of CO as a molecular probe

FTIR spectra of CO adsorbed on Pt/KL catalysts show that the relative band intensities and the total dispersion markedly depend on the catalyst preparation method (ion exchange, incipient wetness impregnation or co-impregnation with KCl). The CO stretching frequency of the dominant band for linear CO is shifted to higher wavenumbers, parallel with the proton concentration in the reduced catalyst, which is derived independently from the intensity of the IR bands of the OH groups. The results are in accordance with the model that electron-deficient platinum particles are platinum-proton adducts.On leave from: N.D. Zelinsky Institute of Organic Chemistry, Academy of Sciences, Moscow, USSR.     

6-Phospho-{beta}-galactosidases of Gram-positive and 6-phospho-{beta}-glucosidase B of Gram-negative bacteria: comparison of structure and function by kinetic and immunological methods and mutagenesis of the lacG gene of staphylococcus aureus

The 6-phospho-ß-galactosidase of Staphylococcus aureus,Lactococcus lactis and Lactobacillus casei and 6-phospho-ßglucosidaseB of Escherichia coli build a subfamily inside a greater enzymefamily, named the glycosal hydrolase family 1, which, hi addition,contains nine ß-glycosidases of different origins.Kinetic and immunological evidence is provided in this reportwhich strengthens the relationship of the four 6-phospho-ß-glycosidases.It is shown that the 6-phospho-ß-galactosidases and6-phospho-ß-glucosidase B are able to split aromaticß-galactoside phosphates and ß-glucosidephosphates. The turnover numbers of hydrolysis of substrateswith different epimerization at C-4 of the glycon vary up to15-fold only. Two polydonal antisera, one derived against thenative 6-phospho-ß-galactosidase from S.aureus andthe other derived against the 6-phospho-ß-glucosidaseB, cross-reacted with both enzymes. Peptides of the proteinswere separated by reverse phase HPLC. The cross-reacting peptideswere sequenced and shown to be localized at almost the sameposition in the aligned primary structures of both enzymes.An insertion of nine amino adds near these antigenic domainsis unique for the 6-phospho-ß-glycosidases and missingwithin the sequences of the ß-glycoside-specific membersof the family. The lacG gene of a 6-phospho-ß-galactosidasenegative S.aureus mutant was doned into E.coli and sequenced.In the totally inactive mutant protein only the glycine at position332 was changed to an arginine. This amino acid is part of thesequence insertion near the antigenic domain reacting with bothantisera. These data support the assumption that the regionis of great importance for the function of the enzymes and thatit is possible it determines the specificity of the phosphorylatedform of the substrates. In addition, the 6-phospho-ß-galactosidaseof S.aureus was modified by sitedirected mutagenesis of thecorresponding lacG gene hi order to replace residues Glul60and Glu375, which were suspected of being involved hi the generalacid catalysis of substrate hydrolysis, with glutamine residues.The mutant protein 160EQ retained some catalytic activity whilethe protein 375EQ was totally inactive. Glu375 is the activesite nudeophile of the 6-phospho-ß-galactosidase ofS.aureus. It is located in the sequence motif ENG where Glu358was identified as the catalytkally active nudeophile hi theß-glucosidase of Agrobacterium.     

Artificial added mass instabilities in sequential staggered coupling of nonlinear structures and incompressible viscous flows

Within this paper the so-called artificial added mass effect is investigated which is responsible for devastating instabilities within sequentially staggered Fluid-structure Interaction (FSI) simulations where incompressible fluids are considered.A discrete representation of the added mass operator M_A is given and ‘instability conditions’ are evaluated for different temporal discretisation schemes. It is proven that for every sequentially staggered scheme and given spatial discretisation of a problem, a mass ratio between fluid and structural mass density can be found at which the coupled system becomes unstable. The analysis is quite general and does not depend upon the particular spatial discretisation schemes used. However here special attention is given to stabilised finite elements employed on the fluid partition. Numerical investigations further highlight the results.     

Induced free liquid surface oscillations in a visco-elastic liquid column due to angular temperature fluctuations

The response due to time-fluctuating temperature distribution applied at the free surface of a viscoelastic cylindrical liquid column with no axial dependency has been determined analytically. The amplitude of the radial- and angular velocity as well as the free liquid surface elevation has been presented as a function of the “reduced” forcing frequency of the oscillatory temperature and has numerically been evaluated for an angular temperature field proportional to sin 2φ. It was found, that visco-elasticity as described by the Maxwell model has quite some influence upon the liquid behavior. It shows for larger relaxation times besides the enlarged resonance peak additional peaks below and above resonance. This is particularly evident inside the liquid column, indicating a more pronounced elastic behavior of the liquid.     

Improvement of the engineering properties of asphalt pavements using a ‘soft’ compaction technique (AMIR-Compaction)

In most countries of the world asphalt pavement of road networks represents the biggest single investment in the transportation system. In Canada, asphalt pavement built over the past decade is worth more than 70 billion dollars (in 1984 dollars). In order to maintain the current road network at the present level of service and prevent it from further deterioration, over 6 billion dollars is needed anually.

Traditionally, asphalt overlays are widely used to rehabilitate existing deteriorated pavements. Unfortunately the new overlays have been observed to fail in a relatively short time. Thus the investment in overlays is lost.

Recent research based on the concept of relative rigidity has indicated that the rapid deterioration of new asphalt overlays is directly related to current construction equipment. It has been shown analytically and experimentally that steel rollers used in compacting the asphalt layer will result in surface cracks during construction. Traffic and environmental influences will accelerate the failure of the new layer.

This paper presents the results of a testing programme evaluating the engineering properties of asphalt slabs compacted by a new method using a compactor termed AMIR. The results showed that the AMIR compactor will prevent the formation of constitution cracks resulting in an increase of indirect tensile strength of 10% and an increase of direct tensile strength of up to 60% when compared to steel roller compacted asphalt slabs.