Hypervelocity impact testing of pressure vessels to simulate spacecraft failure

A series of hypervelocity impact tests are conducted against thin-walled aluminum pressure vessels to investigate failure mechanisms. The vessels, 0.05 and 0.08 inches thick, are constructed to replicate the material properties of the International Space Station (ISS). The vessels are pressurized to simulate the conditions experienced by the habitable modules of the ISS. A test matrix incorporating shielded thin plates, shielded vessels under no pressure and shielded vessels under internal pressure is developed to take advantage of knowledge gained in earlier tests. Given the design parameters of the ISS coupled with the capabilities of light gas gun testing, it is shown that a catastrophic failure due to unzipping is unlikely.     

Directed assembly of functional light harvesting antenna complexes onto chemically patterned surfaces

We report the directed assembly of the photosynthetic membrane proteins LH1 and LH2 isolated from the purple bacterium Rhodobacter sphaeroides onto chemically patterned substrates. Nanoimprint lithography was used to pattern discrete regions of amino-?and fluoro-terminated or poly(ethylene glycol) self-assembled monolayers onto a glass substrate. Densely packed layers of assembled protein complexes were observed with atomic force microscopy. The protein complexes attached selectively to the amino-terminated regions by electrostatic interactions. Spectral images generated with a hybrid scanning probe and fluorescence microscope confirmed that the patterned proteins retained their native optical signatures.     

Development of soluble glasses for biomedical use Part II: The biological response of human osteoblast cell lines to phosphate-based soluble glasses

Soluble glasses are considered to be of potential clinical value in orthopaedic and dental surgery. However, the biological response to these materials is not well understood. To determine the effects of these glasses, two human osteoblast cell lines, MG63 and HOS (TE85), were incubated in vitro in the presence of increasing concentrations of extracts of the glasses. The effects of the extracts on cell growth was measured using the MTT assay and an ELISA assay was used to measure the expression of bone sialoprotein (BSP), osteonectin (ON) and fibronectin (FN), antigens which play a fundamental part in the integrity and function of hard connective tissue. The results showed that the proliferation of the cells was adversely affected only by the more soluble glasses, which also down-regulated the expression of the bone-associated proteins. In contrast, the extract of the glass with the lowest dissolution rate, which contains relatively elevated levels of Ca²⁺, was found to enhance bone cell growth and antigen expression. These findings suggest that the compositions of these glasses at least partly determine the response of cells and thus, that the glasses could be modified to elicit a more optimal biological response and clinical efficacy.     

Tumor efficacy and bone marrow-sparing properties of TER286, a cytotoxin activated by glutathione S-transferase

TER286 is a latent drug activated by human glutathione S-transferase (GST) isoforms P1-1 and A1-1 to produce a nitrogen mustard alkylating agent. M7609 human colon carcinoma, selected for resistance to doxorubicin, and MCF-7 human breast carcinoma, selected for resistance to cyclophosphamide, both showed increased sensitivity to TER286 over their parental lines in parallel with increased expression of GST P1-1. In primary human tumor clonogenic assays, the spectrum of cytotoxic activity observed for TER286 was both broad and unusual when compared to a variety of current drugs. In murine xenografts of M7609 engineered to have high, medium, or low GST P1-1, responses to TER286 were positively correlated with the level of P1-1. Cytotoxicity was also observed in several other cell culture and xenograft models. In xenografts of the MX-1 human breast carcinoma, tumor growth inhibition or regression was observed in nearly all of the animals treated with an aggressive regimen of five daily doses. This schedule resulted in a 24-h posttreatment decline in bone marrow progenitors to 60% of control and was no worse than for a single dose of TER286. These studies have motivated election of TER286 as a clinical candidate.     

Alterations in GABAA receptor alpha 1 and alpha 4 subunit mRNA levels in thalamic relay nuclei following absence-like seizures in rats

Modification of GABAA receptor mRNA levels by seizure activity can regulate general neuronal excitability. The possibility of absence seizure-induced alteration in GABAA receptor alpha 1, alpha 4, beta 2, and gamma 2 subunit gene expression in thalamic relay nuclei was studied in a rat model of absence seizures induced by gamma-hydroxybutyric acid (GHB). We observed a marked increase in alpha 1 mRNA and a corresponding decrease in alpha 4 mRNA in thalamic relay nuclei 2-4 h after the onset of GHB-induced absence seizures (when the seizures were terminating). These changes were selective to these alpha isoforms as neither beta 2 nor gamma 2 mRNA changed following seizures and occurred only in thalamic relay nuclei but not in hippocampus, a structure from which absence seizures do not evolve. The alterations in alpha 1 and alpha 4 mRNA persisted until about 12 h, and by 24 h after the seizure-onset the mRNA levels normalized. Blocking GHB-seizures produced no change in the levels of alpha 1 and alpha 4 mRNA in thalamic relay nuclei, suggesting that seizures themselves were responsible for mRNA alterations. In order to determine if absence seizure-induced changes in alpha 1 and alpha 4 mRNA had any physiological significance, GHB was readministered in rats 6 and 24 h after the onset of seizures. The total duration of GHB-seizures was found to be significantly decreased when GHB was readministered at 6 h but not 24 h after the seizure-onset. These results suggest that absence seizures regulate GABAA receptor alpha 1 and alpha 4 gene expression in thalamic relay nuclei as a compensatory mechanism by which absence seizures are terminated.     

23S rRNA positions essential for tRNA binding in ribosomal functional sites

rRNA plays an important role in function of peptidyl transferase, the catalytic center of the ribosome responsible for the peptide bond formation. Proper placement of the peptidyl transferase substrates, peptidyl-tRNA and aminoacyl-tRNA, is essential for catalysis of the transpeptidation reaction and protein synthesis. In this report, we define a small set of rRNA nucleotides that are most likely directly involved in binding of tRNA in the functional sites of the large ribosomal subunit. By binding biotinylated tRNA substrates to randomly modified large ribosomal subunits from Escherichia coli and capturing resulting complexes on the avidin resin, we identified four nucleotides in the large ribosomal subunit rRNA (positions G2252, A2451, U2506, and U2585) whose modifications prevent binding of a peptidyl-tRNA analog in the P site and one residue (U2555) whose modification interferes with transfer of peptidyl moiety to puromycin. These nucleotides represent a subset of positions protected by tRNA analogs from chemical modification and significantly narrow the number of 23S rRNA nucleotides that may be directly involved in tRNA binding in the ribosomal functional sites.     

Cementless implant composition and femoral stress. A finite element analysis

Proximal atrophy and thigh pain are recognized problems with some cementless femoral stems in total hip arthroplasty. It is thought that reduced femoral stress from alterations in load transfer caused by an intramedullary stem contributes to proximal femoral atrophy. An increase in flexural rigidity and bone stress near the stem tip is thought to contribute to thigh pain. A three-dimensional finite element analysis study was performed to calculate stresses in the proximal femur and bone near the stem tip before and after implantation of a collared, proximally coated, cementless femoral prosthesis. The influence of prosthetic material was examined by changing implant composition from cobalt chrome to titanium alloy and leaving all other parameters constant. Femoral stress was increased twofold immediately below the collar with the titanium implant compared with the cobalt chrome. However, the proximal femoral stress in the titanium implanted model was still 1/10 that in the corresponding region of the unimplanted femur model. At the stem tip, as much as a 30% reduction in femoral stress was seen with the titanium stem compared with the cobalt chrome. These findings suggest biomechanical evidence of an advantage for titanium as an implant material compared with cobalt chrome for cementless femoral stems.     

Glutathione S-transferase of goat lens: evidence for expression of only class mu isoenzymes

Misbisection of lines is thought to represent an attentional bias. When radial lines (intersection of the midsagittal and transverse planes) are presented below eye level, normal subjects are biased toward far peripersonal space in the visual modality and to near peripersonal space in the tactile modality. These errors may be related to a body centered, a retinotopic, or an object centered attentional bias. The purpose of this study was to contrast the body centered and retinotopic-objective centered hypotheses by having 12 normal subjects perform visual and tactile bisections of radial lines that are above and below eye level. The top of the page, which may be defined by retinotopic or object centered coordinates, contains the portion of the line that is most distant from our bodies when the page is below eye level. However, above eye level, the top of a radial line would be the portion of the page that is most proximal to our bodies. We observed that when stimuli are presented below eye level, normal subjects have a visual bias toward far peripersonal space or the top of the page or both, and have a tactile bias in the opposite direction. In the above eye position we found no overall bias in either modality. Because above eye level the body centered bias should have remained the same but the retinotopic or object centered bias should have reversed, our results suggest that the body and object centered or retinotopic biases, which are oriented in opposite directions, nullified each other.