Transient IkappaB kinase activity mediates temporal NF-kappaB dynamics in response to a wide range of tumor necrosis factor-alpha doses.

Dynamic properties of signaling pathways control their behavior and function. We undertook an iterative computational and experimental investigation of the dynamic properties of tumor necrosis factor (TNF)alpha-mediated activation of the transcription factor NF-kappaB. Surprisingly, we found that the temporal profile of the NF-kappaB activity is invariant to the TNFalpha dose.

Mis-specified cells die by an active gene-directed process, and inhibition of this death results in cell fate transformation in Drosophila.

Incorrectly specified or mis-specified cells often undergo cell death or are transformed to adopt a different cell fate during development. The underlying cause for this distinction is largely unknown. In many developmental mutants in Drosophila, large numbers of mis-specified cells die synchronously, providing a convenient model for analysis of this phenomenon.

Dimerization of the major birch pollen allergen Bet v 1 is important for its in vivo IgE-cross-linking potential in mice.

In type I allergy, the cross-linking of membrane IgE on B lymphocytes and of cytophilic IgE on effector cells by their respective allergens are key events. For cross-linking two IgE molecules, allergens need at least two epitopes. On large molecules, these could be different epitopes in a multivalent, or identical epitopes in a symmetrical, fashion.

Assay for the measurement of copeptin, a stable peptide derived from the precursor of vasopressin.

Background: Arginine vasopressin (AVP) is a key regulator of water balance, but its instability makes reliable measurement difficult and precludes routine use. We present a method for quantifying AVP release by use of copeptin, a glycopeptide comprising the C-terminal part of the AVP prohormone.

Methods: We measured copeptin in 50-microL serum and plasma samples from healthy individuals and from critically ill patients with sepsis.

Measurement of midregional proadrenomedullin in plasma with an immunoluminometric assay.

Background: Adrenomedullin (ADM) is a potent vasodilatory peptide, and circulating concentrations have been described for several disease states, including dysfunction of the cardiovascular system and sepsis. Reliable quantification has been hampered by the short half-life, the existence of a binding protein, and physical properties. Here we report the technical evaluation of an assay for midregional pro-ADM (MR-proADM) that does not have these problems.

Role of kinetic effects on the polarization current around a magnetic island.

The polarization current due to a magnetic island rotating in a tokamak plasma is believed to play a central role in the initial evolution of the neoclassical tearing mode. Monte Carlo delta f simulations are performed which also cover a parameter range that is not amenable to analytic treatment but is relevant to experiments. It is shown that the polarization current can change sign when the island rotation frequency is of the order of the ion parallel streaming around the island.

Ternary systems of nonionic surfactant Brij 35, water and various simple alcohols: Structural investigations by small-angle X-ray scattering and dynamic light scattering.

Structural properties of ternary systems composed of nonionic surfactant dodecyl-poly(ethylene oxide-23) ether (C12E23, commercial name: Brij 35), water and various alcohols from ethanol to 1-decanol have been investigated using small-angle X-ray scattering (SAXS) and dynamic light scattering (DLS) techniques. All measurements were performed at the temperature 25 degrees C. SAXS experimental data were put on absolute scale using water as a secondary standard.

Three-dimensional biomechanical properties of the human rectum evaluated with magnetic resonance imaging.

A method to evaluate the three-dimensional (3-D) geometry of the human gastrointestinal wall may be valuable for understanding tissue biomechanics, mechano-sensation and function. In this paper we present a magnetic resonance imaging (MRI) based method to determine rectal geometry and validation of data obtained in three volunteers. A specially designed rectal bag was filled in a stepwise manner while MRI and bag pressure were recorded.

Proteolytic processing pattern of the endothelin-1 precursor in vivo.

Endothelin-1 (ET-1) is a potent vasoconstrictor, which has been implicated in diseases involving dysfunctions of the cardiovascular system. For the biogenesis of ET-1, a larger precursor peptide (proET-1) is cleaved at two sites to give rise to bigET-1, which is subsequently cleaved to generate mature ET-1. In the present study, we investigated, which other peptides are derived from proET-1 in vivo.

Serum neopterin for early assessment of severity of severe acute respiratory syndrome.

Neopterin and C-reactive protein (CRP) concentrations were determined in serum samples from 129 severe acute respiratory syndrome (SARS) patients and 156 healthy blood donors. In the patients with confirmed SARS, an early neopterin elevation was detected already at the day of onset of symptoms and rose to a maximum level of 45.0 nmol/L 3 days after the onset.

Copeptin, a stable peptide derived from the vasopressin precursor, is elevated in serum of sepsis patients.

Vasopressin is one of the key regulators of the body's water and solute balance. When this balance is pathologically disturbed, determination of serum vasopressin concentrations might be a helpful tool for guiding therapy. However, due to its instability and considerable association to platelets, reliable measurement of circulating vasopressin is difficult to achieve, if at all.

Release of the mitochondrial enzyme carbamoyl phosphate synthase under septic conditions.

To identify sepsis-related dysregulations of protein expression in the liver, we used a baboon model of acute endotoxemia and performed comparative proteome analysis. Treatment with lipopolysaccharide (LPS) was followed by an early but long-lasting (5-48 h) generation of N-terminal fragments of carbamoyl phosphate synthase-1 (CPS-1), an abundant enzyme of the hepatic urea cycle, which is normally located in the mitochondrial matrix. In addition, we developed a new sandwich immunoassay to determine circulating CPS-1 in human and baboons.

Decreased serum amyloid beta(1-42) autoantibody levels in Alzheimer's disease, determined by a newly developed immuno-precipitation assay with radiolabeled amyloid beta(1-42) peptide.

Background: Autoantibodies against amyloid beta (A beta) peptide found in patients with Alzheimer's disease (AD) also occur naturally in the general population independently of the cognitive status.

Methods: We compared serum A beta(1-42) autoantibody levels (A beta(1-42)-AL) of 96 AD patients and 30 healthy elderly control subjects (HC), assessing their diagnostic value for AD with a newly developed immunoprecipitation assay with radiolabeled A beta(1-42) peptide.

Results: We found a highly significant decrease of A beta(1-42)-AL in AD patients (p = .

The CARD-carrying caspase Dronc is essential for most, but not all, developmental cell death in Drosophila.

The initiator caspase Dronc is the only Drosophila caspase that contains a caspase activation and recruitment domain (CARD). Although Dronc has been implicated as an important effector of apoptosis, the genetic function of dronc in normal development is unclear because dronc mutants have not been available. In an EMS mutagenesis screen, we isolated four point mutations in dronc that recessively suppress the eye ablation phenotype caused by eye-specific overexpression of hid.

Genetic control of programmed cell death in Drosophila melanogaster.

Apoptosis is a genetically controlled form of cell death that is an important feature of animal development and homeostasis. The genes involved in the control and execution of apoptosis are conserved throughout evolution. However, the actual molecular mechanisms used by these genes vary from species to species.

The decrease in gamma-tocopherol in plasma and lipoprotein fractions levels off within two days of vitamin E supplementation.

The effects of vitamin E supplementation on alpha- and gamma-tocopherol concentrations were studied in plasma and lipoprotein fractions of five healthy volunteers taking 1000 IU/day of RRR alpha-tocopherol for 4 days. Although plasma alpha-tocopherol increased, gamma-tocopherol decreased. Compared with baseline, gamma-/alpha-tocopherol ratios decreased from 48 h onward (P < 0.

Pro-atrial natriuretic peptide is a prognostic marker in sepsis, similar to the APACHE II score: an observational study.

Introduction: Additional biomarkers in sepsis are needed to tackle the challenges of determining prognosis and optimizing selection of high-risk patients for application of therapy. In the present study, conducted in a cohort of medical intensive care unit patients, our aim was to compare the prognostic value of mid-regional pro-atrial natriuretic peptide (ANP) levels with those of other biomarkers and physiological scores.

Methods: Blood samples obtained in a prospective observational study conducted in 101 consecutive critically ill patients admitted to the intensive care unit were analyzed.

Acute exacerbation of systemic scleroderma in Borrelia burgdorferi infection.

In recent years a possible aetiological connection between skin sclerosis and an infection with Borrelia burgdorferi has been discussed, but this association has not yet been reported for systemic scleroderma. Several treatment modalities are suggested for systemic scleroderma, but no treatment has yet been found to alter the overall course of the disease. This report describes a 61-year-old woman with Raynaud's phenomenon, nail-fold changes and circulating anticentromere antibodies, who showed an abrupt onset of erythemas and doughy swellings involving the face and upper trunk, followed by thickening and induration of the skin mimicking diffuse systemic scleroderma.

TraA and its N-terminal relaxase domain of the Gram-positive plasmid pIP501 show specific oriT binding and behave as dimers in solution.

TraA is the DNA relaxase encoded by the broad-host-range Grampositive plasmid pIP501. It is the second relaxase to be characterized from plasmids originating from Gram-positive organisms. Full-length TraA (654 amino acids) and the N-terminal domain (246 amino acids), termed TraAN246, were expressed as 6xHis-tagged fusions and purified.

Running throughout pregnancy: effect on placental villous vascular volume and cell proliferation.

Many studies have documented that placental development is altered by a variety of environmental factors which alter placental bed blood flow and/or oxygen delivery. One of these is sustained weight-bearing exercise. The purpose of this investigation was to examine the effects of running throughout pregnancy on villous vascular development and cell proliferation by testing the null hypothesis that continuing a regular running regimen throughout pregnancy has no effect on villous vascular volume or cell proliferation at term.

Identification of an Adrenomedullin precursor fragment in plasma of sepsis patients.

Adrenomedullin and PAMP are potent vasodilatory peptides derived from a common larger precursor peptide. Elevation of circulating levels of both peptides has been described for diseases involving dysfunction of the cardiovascular system. However, the reliable quantification has been hampered by their short half-life times and - as known for Adrenomedullin -- the existence of a binding protein.

Interaction of PSD-95 with potassium channels visualized by fluorescence lifetime-based resonance energy transfer imaging.

Resonance energy transfer (RET) has been extensively used to estimate the distance between two different fluorophores. This study demonstrates how protein-protein interactions can be visualized and quantified in living cells by time-correlated single-photon counting (TCSPC) imaging techniques that exploit the RET between appropriate fluorescent labels. We used this method to investigate the association of the potassium inward rectifier channel Kir2.

Multi-dimensional time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to detect FRET in cells.

We present a novel, multi-dimensional, time-correlated single photon counting (TCSPC) technique to perform fluorescence lifetime imaging with a laser-scanning microscope operated at a pixel dwell-time in the microsecond range. The unsurpassed temporal accuracy of this approach combined with a high detection efficiency was applied to measure the fluorescent lifetimes of enhanced cyan fluorescent protein (ECFP) in isolation and in tandem with EYFP (enhanced yellow fluorescent protein). This technique enables multi-exponential decay analysis in a scanning microscope with high intrinsic time resolution, accuracy and counting efficiency, particularly at the low excitation levels required to maintain cell viability and avoid photobleaching.

Lineage-specific imprinting and evolution of the zinc-finger gene ZIM2.

We have carried out an in-depth comparative analysis of a 100-kb genomic interval containing two imprinted genes, PEG3 and ZIM2, using sequences derived from human, mouse, and cow. In all three mammals, ZIM2 is located at a similar genomic distance and in the same orientation relative to PEG3, indicating the basic structural conservation of this imprinted locus. However, several lineage-specific changes have occurred that affect the exon structure and imprinting status of ZIM2.

Does 3-D sonography bring any advantage to noninvasive breast diagnostics?

The aim of this prospective trial was to evaluate if 3-D ultrasound (US) brings any advantage to breast diagnostics. A total of 65 women with breast lesions (42 malignant, 23 benign) were examined preoperatively with 2-D and 3-D US. The impact of the use of the 3-D coronal plane for the visualization of the infiltrative zone in comparison to 2-D US was evaluated.