Intracellular transport of class I MHC molecules in antigen processing mutant cell lines

Intracellular transport and stability of class I MHC glycoproteins depends on the assembly of H chain, beta 2-microglobulin, and peptide. The Ag processing mutant cell lines T2 and RMA-S have defects in peptide loading of class I, resulting in reduced cell surface expression of class I molecules. Expression of class I molecules in the murine cell line RMA-S can be induced at 26 degrees C, suggesting that they are transported to the cell surface, but are unstable. However, most human class I molecules in T2 are poorly expressed at the cell surface, even at 26 degrees C. To directly compare the transport of human and mouse alleles in RMA-S and T2, the human alleles HLA-A2, A3, and B27 were transfected into RMA-S along with human beta 2-microglobulin, and the mouse alleles H-2Kb and Db were transfected into T2. Surface expression of HLA-A3 and B27 in RMA-S remained less than 10% of wild-type levels at 26 degrees C. H-2Kb and Db in both cell lines, however, were expressed at 20 to 30% wild-type levels at 37 degrees C and could be induced to wild-type levels at 26 degrees C or with peptides. The selective expression of murine class I glycoproteins at the cell surface of T2 is not because of their greater stability when associated with human beta 2m, since H-2Kb and Db H chain/human beta 2m complexes dissociate more rapidly in vitro than HLA-A3 and B27 complexes. These results suggest that the difference in transport between human and mouse class I in T2 reflects a fundamental structural property of the class I glycoproteins.     

Separating the effects of alertness from the effects of encoding in a pitch-imagery task

This article describes a general approach entitled Wizard of Oz hypnotherapy for use with 8- to 13-year-old children who are generally difficult to treat because of their intermittent inattentive, impulsive, and oppositional characteristics. We have utilized this approach as an adjunct to cognitive-behavioral therapy individually and within a small group format. The Wizard of Oz metaphor is integrated into a hypnotherapy approach designed to facilitate personal problem-solving, improve self-regulation skills, and enhance self-esteem/efficacy. As an adjunct to cognitive-behavioral therapy, this procedure improves the therapeutic alliance, heightens young clients' interest in therapy procedures, and improves overall cooperation with treatment. The therapeutic and hypnotic characteristics that may contribute to this technique's effectiveness are discussed.     

Age-related changes in hepatic and splenic insulin receptors and serum insulin and glucose levels in inbred mice

A 62-year-old man and 26-year-old man with re-expansion pulmonary edema (RPE) after thoracic drainage as a treatment for pneumothorax are presented. Blood cell counting and biochemical serum analysis were performed throughout their treatment in both patients, and biochemical sputum analysis was in one patient. The results showed transient marked leukocytosis just after RPE. Total protein and albumin concentrations of sputum approximated to those of serum. The above results suggested that RPE is based on pulmonary microvascular injury, which may introduce leukocytosis.     

Treponema denticola outer membrane inhibits calcium flux in gingival fibroblasts

Treponema denticola is a cultivable oral spirochete which perturbs the cytoskeleton in cultured cells of oral origin, but intracellular signalling pathways by which it affects actin assembly are largely unknown. As the outer membrane (OM) of Treponema denticola disrupts actin-dependent processes that normally require precise control of intracellular calcium, we studied the effects of an OM extract on internal calcium release, ligand-gated and calcium release-activated calcium channels, and related mechanosensitive cation fluxes in human gingival fibroblasts (HGF). Single-cell ratio fluorimetry demonstrated that in resting cells loaded with Fura-2, baseline intracellular Ca2+ concentration ([Ca2+]i) was not affected by treatment with OM extract, but normal spontaneous [Ca2+]i oscillations were dramatically increased in frequency for 20 to 30 min followed by complete blockade. OM extract inhibited ATP-induced and thapsigargin-induced release of calcium from intracellular stores by 40 and 30%, respectively. Addition of Ca2+ to the extracellular pool following depletion of intracellular Ca2+ by thapsigargin and extracellular Ca2+ by EGTA yielded 59% less replenishment of [Ca2+]i in OM extract-treated than in control HGF. In cells loaded with collagen-coated ferric oxide beads to stimulate integrin-dependent calcium release, baseline [Ca2+]i was nearly doubled but was not significantly different in control and OM extract-treated cells. Magnetically generated tensile forces on the beads induced >300% increases of [Ca2+]i above baseline. Cells preincubated with OM extract exhibited dose-dependent and time-dependent reductions in stretch-induced [Ca2+]i transients, which were due to neither loss of beads from the cells nor cell death. The T. denticola OM inhibitory activity was eliminated by heating the OM extract to 60 degrees C and by boiling but not by phenylmethylsulfonyl fluoride treatment. Thus nonlipopolysaccharide, nonchymotrypsin, heat-sensitive protein(s) in T. denticola OM can evidently inhibit both release of calcium from internal stores and uptake of calcium through the plasma membrane, possibly by interference with calcium release-activated channels.     

Agrobacterium tumefaciens transformation of the radiation hypersensitive Arabidopsis thaliana mutants uvh1 and rad5

The Arabidopsis thaliana mutants uvh1 and rad5, originally identified as radiation hypersensitive, were reported to be deficient in T-DNA integration based on the relative efficiencies of stable transformation and T-DNA transfer. We reassessed these mutants for susceptibility to transformation by Agrobacterium tumefaciens. The mutant rad5 showed a significant reduction in the efficiency of transient as well as stable transformation, compared with its wild-type progenitor. These data indicate that rad5 is blocked at a step in the transformation process prior to T-DNA integration. We additionally found, using both an in vitro root inoculation and an in vivo flower bolt inoculation assay, that the mutant uvh1 is as susceptible to A. tumefaciens-mediated transformation as is its wild-type progenitor, C10.     

Optimization of the deposited power distribution inside a layered lossy medium irradiated by a coupled system of concentrically placed waveguide applicators

A method is proposed for controlling the deposited power distribution in a layered cylindrical lossy model, irradiated by a phased-array hyperthermia system consisting of four waveguide applicators. A rigorous electromagnetic model of the heated tissue, which takes into account coupling phenomena between system elements, is used for predicting the electric field at any point inside tissue. The relative amplitudes and relative phases of the array elements are optimized in order to attain desired specific absorption rate (SAR) distributions inside and outside malignant tissues. A constrained nonlinear optimization problem is solved by using the penalty function method and the resulting unconstrained minimization of the penalty function is carried out by the downhill simplex method. Two practical phased-array hyperthermia systems have been studied and numerical results are presented.