An in vitro model for toxin-mediated vascular leak syndrome: ricin toxin A chain increases the permeability of human endothelial cell monolayers

Vascular leak syndrome (VLS) is the dose-limiting toxicity observed in clinical trials of immunotoxins containing ricin toxin A chain (RTA). RTA itself is thought to cause VLS by damaging vascular endothelial cells, but the exact mechanism remains unclear. This is partially due to the paucity of appropriate models. To study VLS, we developed an in vitro model in which human umbilical vein-derived endothelial cells were first grown to confluence on microporous supports and then cultured under low pressure in the presence or absence of RTA. Endothelial cell barrier function was assessed by measuring the volume of fluid that passed through each monolayer per unit time. We found that RTA significantly increased monolayer permeability at times and concentrations consistent with the onset of VLS in patients treated with RTA-based immunotoxins. Scanning electron microscopy showed that intercellular gaps formed in endothelial monolayers exposed to RTA. Intercellular gap formation followed endothelial cell death caused by the enzymatic activity of RTA. We conclude that RTA is directly toxic to endothelial cells in vitro and speculate that this contributes to VLS in vivo.     

The interaction of peripheral blood leukocytes with alpha1-acid glycoprotein, its carbohydrate chains and neoglycoconjugates

The interaction of human peripheral blood leukocytes with alpha 1-acid glycoprotein (AGP), its glycoforms as well as neoglyco-conjugates representing carbohydrate chains of AGP or its fragments was studied by flow cytometry. It was shown that the main target cells for AGP as well as for conjugates of its carbohydrate chains with polyacrylamide (PAA) are monocytes and polymorphonuclear leukocytes but not lymphocytes. The interaction of AGP with monocytes and granulocytes are mediated by its carbohydrate chains: the binding of AGP with cells was inhibited by AGP, AGP oligosaccharides as well as conjugates of oligosaccharides and its fragments with PAA. The data obtained show the existence of monocyte (and granulocyte) receptors which interact with complex type sialooligosaccharides of AGP.     

Solution conformation of the Pseudomonas syringae MSU 16H phytotoxic lipodepsipeptide Pseudomycin A determined by computer simulations using distance geometry and molecular dynamics from NMR data

Pseudomycin A is a cyclic lipodepsinonapeptide phytotoxin produced by a strain of the plant pathogenic bacterium Pseudomonas syringae. Like other members of this family of bacterial metabolites, it is characterised by a fatty acylated cyclic peptide with mixed chirality and lactonic closure. Several biological activities of Pseudomycin A are lower than those found for some of its congeners, a difference which might depend on the diverse number and distribution of charged residues in the peptide moiety. Hence, it was of interest to investigate its conformation in solution. After the complete interpretation of the two-dimensional NMR spectra, NOE data were obtained and the structure was determined by computer simulations, applying distance geometry and molecular dynamics procedures. The conformation of the large ring of Pseudomycin A in solution includes three rigid structural regions interrupted by three short flexible regions that act as hinges. The overall three-dimensional structure of the cyclic moiety is similar to that of previously studied bioactive lipodepsinonapeptides produced by other pseudomonads.     

Myxomas of the mandible and maxilla

Two cases of congenital absence of a cervical vertebral pedicle are reported. This condition includes hypoplasia of the ipsilateral posterior arch and may predispose to spinal cord injury. The radiographic and computed tomography (CT) findings are reviewed, and the importance of the ipsilateral oblique radiographic view and of CT in diagnosis is stressed.     

A monomial-based method for solving systems of non-linear algebraic equations

A monomial-based method for solving systems of algebraic non-linear equations is presented. The method uses the arithmetic-geometric mean inequality to construct a system of monomial equations that approximates the system of non-linear equations. A change of variables transforms the monomial system into a system of linear equations, which is readily solved. Special properties of the monomial method are identified and their significance is discussed. Invariance properties of the monomial method produce a built-in, self-adjusting scaling of the variables and equilibration of the equations of the linear system. Other special properties can lead to useful bounds on, and invariances of, the conditioning of the linear system. An invariance to uniform scaling is responsible for extremely rapid convergence to the equation surfaces in the initial iterations. An invariance to multiplication of the algebraic equations by a certain class of functions leads to a useful insensitivity to form of the algebraic system. Insensitivity of the monomial method to solutions with negative components avoids meaningless solutiuons of the algebraic system that appear as undesirable by-products of the formulation. A practical engineering design problem is solved to demonstrate the special properties of the monomial method.     

Jitter reduction through feedback for picosecond pulsed InGaAsPlasers

Following a comparison of picosecond-pulse generation techniques, feedback schemes are reported for the generation of picosecond pulse trains with improved jitter for both multicontact and conventional single-contact InGaAsP-InP lasers. Subpicosecond jitter is achieved for Q-switched laser sources using a novel optoelectronic feedback scheme. The use of resonant electrical feedback is shown to improve the timing jitter of gain-switched pulses by up to six times. Pulse-to-pulse timing jitter as low as 250 fs is demonstrated for a hybrid of optical and electrical feedback schemes. Limits for timing jitter in diode lasers are established for optoelectronic, electrical, and optical feedback schemes, and the key picosecond pulse generation schemes are compared in terms of timing jitter for the first time