ß1-Integrin cytoskeletal signaling regulates sensory neuron response to matrix dimensionality.

Neuronal differentiation, pathfinding and morphology are directed by biochemical cues that in vivo are presented in a complex scaffold of extracellular matrix. This microenvironment is three-dimensional (3D) and heterogeneous. Therefore, it is not surprising that more physiologically-relevant cellular responses are found in 3D culture environments rather than on two-dimensional (2D) flat substrates. One key difference between 2D and 3D environments is the spatial arrangement of cell-matrix interactions. Integrins and other receptor proteins link the various molecules presented in the extracellular environment to intracellular signaling cascades and thus influence a number of neuronal responses including the availability and activation of integrins themselves. We have previously reported that a 3D substrate induces an important morphological transformation of embryonic mouse dorsal root ganglion (DRG) neurons. Here, we investigate the hypothesis that ß1-integrin signaling via focal adhesion kinase (FAK) and the RhoGTPases Rac and Rho influences neuronal morphology in 2D vs 3D environments. We report that ß1-integrin activity and FAK phosphorylation at tyrosine 397 (FAKpY397) are linked to neuronal polarization as well as neurite outgrowth and branching. Rac and Rho expression are decreased in 3D vs 2D culture but not correlated with ß1-integrin function. These results suggest that proper ß1-integrin activity is required for the elaboration of physiologic DRG morphology and that 3D culture provides a more appropriate milieu to the mimic in vivo scenario. We propose that neuronal morphology may be directed during development and regeneration by factors that influence how ß1-integrin, FAK and RhoGTPase molecules integrate substrate signals in the 3D microenvironment.

Inhibition by albumin of merocyanine 540-mediated photosensitization of platelets and viruses.

The effect of the photosensitizer merocyanine 540 (MC 540) on platelets and on three marker viruses was examined to assess its potential in reducing virus transmission by blood products. The results demonstrated several deleterious effects of MC 540 (4-24 micrograms/mL) on platelet morphology and function in both the absence and presence of light (450-600 nm). Treatment of washed platelets with MC 540 in the dark resulted in a significant release of serotonin in the absence of added agonist, as well as a diminished response to thrombin as measured in vitro. In addition, photosensitization caused spontaneous platelet aggregation and release of 92 percent of the releasable serotonin without the addition of an agonist. Because photo-treatment of blood products is likely to be performed in a protein-rich medium, the influence of albumin on the phototoxic effects on platelets was assessed. Albumin added to the suspension medium at concentrations greater than or equal to 1.0 percent protected the platelets against the effects of MC 540 in the dark, whereas 5-percent albumin was required for protection against the phototoxic effects of MC 540 on the platelet response to thrombin. The antiviral activity of MC 540 and light was examined by using the lipid-containing viruses herpes simplex virus (HSV) and bacteriophages phi 6 and PM2. Of the lipid-enveloped viruses, HSV was 25 times more photosensitive to MC 540 than was phi 6 (15 micrograms/mL). PM2, which has an internal lipid layer, was almost 300 times less sensitive to MC 540 and light than was HSV.(ABSTRACT TRUNCATED AT 250 WORDS)

Salmonella typhi vaccine strain in vitro; low infectivity in human cell line U937.

Salmonella typhi strain Ty21a has been used for live oral vaccine. The infectivity of Ty21a, in comparison with S. typhi Ty2, was evaluated using the human monocyte-macrophage cell line U937. Assays were performed by quantitative microscopy and viable count technique. Ty2 infected approximately 100% of the cells, multiplied extensively within these cells and caused cell death. The same dose of Ty21a infected only about 15% of the cells, resulting in a low number of intracellular bacilli and cell survival. The use of gentamicin in the test confirmed intracellular multiplication of Ty2 but not Ty21a. The system described may be suitable as a test system for characterization of the degree of virulence of Ty21a and other live, oral typhoid vaccines.

Influence of high pass filtering on the intelligibility of amplitude-compressed speech.

Monosyllabic triplet word intelligibility scores were obtained from normal-hearing and hearing-impaired, loudness-recruiting subjects under two experimental conditions: (1) high-pass (1200 Hz)-filtered, linear amplification, and (2) high-pass (1200 Hz)-filtered, compression amplification using input-to-output ratios of 5:1 and 20:1. Test materials were administered at increased sensation levels of 0, 10, 20, and 30 dB. In general, speech intelligibility was slightly enhanced for normal and hearing-impaired listeners, but only at lower sensation levels. Moreover, the improvement was observed only under the filtered, compression amplification condition for both groups. No important differences were observed between the two compression ratios used. This compression advantage may or may not be observed in clinical hearing aid evaluations.