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Summary: Vaccination against meningococcal serogroup B is recommended for patients with a complement deficiency; however, although immunogenicity in this patient group has been shown, efficacy has not yet been established. In this study, we collected serum from children with a complement deficiency in the alternative pathway or in late terminal pathway before and after vaccination with multi‐component meningococcal serogroup B (MenB)‐4C. MenB‐4C is a multi‐component, protein‐based vaccine against MenB consisting of factor H‐binding protein, Neisserial heparin‐binding protein, Neisserial adhesion A and outer membrane vesicles containing Porin A. We assessed the vaccine immunogenicity and vaccine‐mediated protection by a whole cell enzyme‐linked immunosorbent assay with Neisseria meningitidis serogroup B strains H44/76, 5/99 and NZ98/254, which shows that vaccination induced antibody titers against meningococcus. We show that the classical serum bactericidal activity assay with exogenous serum indicates the presence of vaccine‐induced antibodies and capacity to activate complement‐mediated pathogen lysis. However, in children with a late terminal pathway deficiency, no complement‐mediated pathogen lysis was observed when autologous serum was applied in the serum bactericidal activity assay, demonstrating a lack of serum bactericidal activity in children with complement deficiencies. However, MenB‐4C vaccination still induced effective complement‐dependent opsonophagocytic killing against N. meningitidis serogroup B in reconstituted whole blood with autologous serum from children with an alternative pathway or late terminal pathway deficiency. These findings support the recommendation to vaccinate all complement‐deficient children against MenB. [ABSTRACT FROM AUTHOR]

We report a polarized fluorescence recovery after photobleaching (pFRAP) method to measure the rotational dynamics of fluorescent colloids over a wide dynamic range. The method is based on the polarization anisotropy in the fluorescence intensity, generated by bleaching of fluorescently labeled particles with an intense pulse of linearly polarized laser light. The rotational mobilities of the fluorescent particles can be extracted from the relaxation kinetics of the postbleach fluorescence polarization anisotropy. Our pFRAP setup has access to correlation times over a range of time scales from tens of microseconds to tens of seconds, and is highly sensitive, so very low concentrations of labeled particles can be probed. We present a detailed description of the theoretical background of pFRAP. The performance of the equipment is demonstrated for fluorescent colloidal silica spheres, dispersed in pure solvents as well as in fd-virus suspensions. © 2004 American Institute of Physics. [ABSTRACT FROM AUTHOR]

A sensitive dielectric spectroscopy setup is built to measure the response of nanoparticles dispersed in a liquid to an alternating electric field over a frequency range from 10-2 to 107 Hz. The measured complex permittivity spectrum records both the rotational dynamics due to a permanent electric dipole moment and the translational dynamics due to net charges. The setup consists of a half-transparent capacitor connected in a bridge circuit, which is balanced on pure solvent only, using a software-controlled compensating voltage. In this way, the measured signal is dominated by the contributions of the nanoparticles rather than by the solvent. We demonstrate the performance of the setup with measurements on a dispersion of colloidal CdSe quantum dots in the apolar liquid decalin. [ABSTRACT FROM AUTHOR]

A sensitive balanced differential transformer was built to measure complex initial parallel magnetic susceptibility spectra in the 0.01–1000 Hz range. The alternating magnetic field can be chosen sufficiently weak that the magnetic structure of the samples is only slightly perturbed and the low frequencies make it possible to study the rotational dynamics of large magnetic colloidal particles or aggregates dispersed in a liquid. The distinguishing features of the setup are the novel multilayered cylindrical coils with a large sample volume and a large number of secondary turns (55 000) to measure induced voltages with a good signal-to-noise ratio, the use of a dual channel function generator to provide an ac current to the primary coils and an amplitude- and phase-adjusted compensation voltage to the dual phase differential lock-in amplifier, and the measurement of several vector quantities at each frequency. We present the electrical impedance characteristics of the coils, and we demonstrate the performance of the setup by measurement on magnetic colloidal dispersions covering a wide range of characteristic relaxation frequencies and magnetic susceptibilities, from χ≈-10-5 for pure water to χ>1 for concentrated ferrofluids. [ABSTRACT FROM AUTHOR]

Abstract: The regulatory role of viruses on population dynamics of the prymnesiophyte Phaeocystis globosa was studied during a mesocosm experiment in relation to growth and loss by microzooplankton grazing and cell lysis. The mesocosms were conducted under varying light conditions (20 and 150μmol photons m−2 s−1) and nutrient regime (inorganic nitrogen to phosphorus ratios of 4, 16 and 44). Overall, viruses infecting P. globosa (PgV) were found to be an important cause of cell lysis (30–100% of total lysis) and a significant loss factor (7–67% of total loss). We demonstrate that the morphology of P. globosa cells (solitary versus colonial) differently regulated viral control of P. globosa bloom formation. Reduced irradiance (20μmol photons m−2 s−1) was provided for 11 days to select for the solitary cell morphotype. Viruses were able to restrict P. globosa bloom formation even after irradiance became saturating again (150μmol photons m−2 s−1). Saturating light conditions from the start of the experiment allowed colony formation and because the colony-morphotype acted as a mechanism reducing viral infection bloom formation succeeded. Nutrient depletion, however, affected specifically the colonies that disintegrated while releasing single cells. Virus infection of these solitary cells resulted in the termination of the bloom. The nature of phytoplankton growth-limiting nutrient (nitrate and/or orthophosphate) did not seem to noticeably affect the level of viral control. [Copyright &y& Elsevier]

Neisseria meningitidis expresses a range of lipooligosaccharide (LOS) structures, comprising of at least 13 immunotypes (ITs). Meningococcal LOS is subject to phase variation of its terminal structures allowing switching between ITs, which is proposed to have functional significance in disease. The objectives of this study were to investigate the repertoire of structures that can be expressed in clinical isolates, and to examine the role of phase-variable expression of LOS genes during invasive disease. Southern blotting was used to detect the presence of LOS biosynthetic genes in two collections of meningococci, a global set of strains previously assigned to lineages of greater or lesser virulence, and a collection of local clinical isolates which included paired throat and blood isolates from individual patients. Where the phase-variable genes lgtA, lgtC or lgtG were identified, they were amplified by PCR and the homopolymeric tracts, responsible for their phase-variable expression, were sequenced. The results revealed great potential for variation between alternate LOS structures in the isolates studied, with most strains capable of expressing several alternative terminal structures. The structures predicted to be currently expressed by the genotype of the strains agreed well with conventional immunotyping. No correlation was observed between the structural repertoire and virulence of the isolate. Based on the potential for LOS phase variation in the clinical collection and observations with the paired patient isolates, our data suggest that phase variation of LOS structures is not required for translocation between distinct compartments in the host. [Copyright &y& Elsevier]

Atomic force microscopy (AFM) is used to study the size, shape, and polydispersity of a variety of magnetic and nonmagnetic model colloids, previously imaged by transmission electron microscopy (TEM) only. Both height and phase images are analyzed and special attention is given to 3D morphology and softness of particles, as well as structures and presence of secondary components in the colloid, difficult to investigate with TEM. Several methods of tip characterization followed by deconvolution were applied in order to improve the accuracy of lateral diameter determination. In the case of magnetite particles dispersed in conventional ferrofluids, we explore both experimentally and theoretically the possibility of using magnetic force microscopy (MFM). We propose and discuss several models which allow to estimate the magnetic moment of a single domain superparamagnetic sphere using MFM, which cannot be done with other techniques; alternatively the tip magnetization can be determined. [Copyright &y& Elsevier]

This paper presents a technical description and some preliminary results of a relatively large experimental tidal mud-flat ecosystem, constructed in parallel set-ups in the experimental aquarium of the Nederlands Instituut voor Onderzoek der Zee (NIOZ). The system was build in 1975, filled with sea-water and natural sediment from a typical Arenicola marina habitat. After introducing micro-phytobenthos and juvenile A. marina, the development in both systems, without correcting interventions, were followed for about two years. Studies on nutrient concentrations and organic matter, primary productivity, fluctuations in biomass, density of secondary producers and activity of micro-organisms reveal the systems to be self-pertaining and fairly stable. The study of assemblages of hardy inhabitants of the stressful intertidal environment seems to be a promising starting point for further micro-system research. [ABSTRACT FROM AUTHOR]

To generate hypotheses about how physicians make difficult clinical decisions, we analyzed transcripts of the "thinking aloud" behavior of expert clinicians making a testing or treatment decision with an uncertain diagnosis. We compared the clinicians' reasoning with a decision analysis of the same problem. The experts did not formulate a global outline of their decision, but chained together a sequence of decisions based on available and incomplete information. Despite effective and efficient problem solving, the clinicians used numeric terms only as symbolic representations of likelihood, used limited information in choosing among alternatives, and dismissed the possibility that a less conventional strategy, empiric therapy, might yield equivalent outcome. We describe cognitive problem-solving strategies and knowledge representations that permit persons to make successful decisions despite limited processing resources. The same cognitive procedures probably contribute to observed errors in decision-making under uncertainty. [ABSTRACT FROM AUTHOR]

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The sensitivity of an imperfectly balanced impedance bridge is limited by the remaining offset voltage. Here, we present a procedure for offset reduction in impedance measurements using a lock-in amplifier, by applying a complex compensating voltage external to the bridge. This procedure takes into account instrumental damping and phase shifting, which generally occur at the high end of the operational frequency range. Measurements demonstrate that the output of the circuit rapidly converges to the instrumentally limited noise at any frequency. [ABSTRACT FROM AUTHOR]