Apoptosis in interface membranes of aseptically loose total hip arthroplasty

The terminal events leading to periprosthetic osteolysis are multifactorial in nature and modulation of this process after the stage of osteolytic mediator release has been futile. Recently, the demonstration of the ability of bisphosphonates to inhibit bone resorption that is mediated by particle-stimulated macrophages and their induction of osteoclast apoptosis suggests a potent area for modulation of osteolysis at the prosthesis-bone interface. The purpose of this study was to determine the mode of cell death that occurs at the osteolytic interface of failed total hip arthroplasty (THA). TUNEL staining, DNA laddering, and immunodetection of poly(ADP-ribose)polymerase (PARP) protein were used to identify the presence of apoptosis in interface membranes from 25 patients aged 28–88 years old (mean, 58 years) harvested at the time of hip revision surgery. Our results demonstrated positive TUNEL stain in 100% of specimens with an average 37% of cells (range 12–60%) positively stained for TUNEL whereas less than 8% of control tissue cells showed positive staining. DNA laddering, a characteristic feature of apoptotic cells, was observed in 82% (28/34) of specimens studied at both the acetabular and femoral side of aseptically loose THAs. No laddering was observed in control tissues. Finally, using Western blot analysis, we observed the appearance of the 89 kDa PARP fragment associated with apoptosis in 92% of specimens (30/33). Our results demonstrate the presence of apoptotic cell death in interface membranes of THAs suggesting that apoptosis-related events are indeed associated with periprosthetic osteolysis and could serve as a specific target point for therapeutic modulation. © 2001 Kluwer Academic Publishers     

What shadows reveal about object structure

In a scene observed from a fixed viewpoint, the set of shadow boundaries in an image changes as a point light source (nearby or at infinity) assumes different locations. We show that for any finite set of point light sources illuminating an object viewed under either orthographic or perspective projection, there is an equivalence class of object shapes having the same set of shadows. Members of this equivalence class differ by a four-parameter family of projective transformations, and the shadows of a transformed object are identical when the same transformation is applied to the light source locations. Under orthographic projection, this family is the generalized bas-relief (GBR) transformation, and we show that the GBR transformation is the only family of transformations of an object's shape for which the complete set of imaged shadows is identical. Finally, we show that given multiple images under differing and unknown light source directions, it is possible to reconstruct both an object's surface and the light source locations up to this family of transformations from the shadows alone.     

Desorption-ionization mass spectrometry using deposited nanostructured silicon films

We present a method for desorption ionization on silicon based on novel column/void-network-deposited silicon thin films. A number of different peptides and proteins in the < or = 6000 Daltons range are analyzed by time-of-flight mass spectrometry in this demonstration of our approach. A variety of sample preparation conditions, including the use of chemical additives, surface treatments, and sample purification are used to show the potential of mass analysis using deposited column/void-network silicon films for high throughput proteomic screening.     

Application of the ART-2a algorithm to laser ablation aerosol mass spectrometry of particle standards

Single-particle mass spectrometers are now commonly used to analyze atmospheric particles and generate tens of thousands of spectra from typical measurement campaigns. The ART-2a spectrum algorithm has been used to classify these spectra. In this work, we generate a range of particles that are models of those that are common in the atmosphere. A single-particle mass spectrometer is used to analyze these known particles, and the spectra are classified using ART-2a. The optimum vigilance parameter is approximately 0.5 while the optimum learning rate is approximately 0.05. The classifications elucidate limitations in generation of test particles, their analysis by single-particle techniques, and their classification by ART-2a.     

Cytotoxicity of triorganophosphinegold(i) complexes of thiobenzoate

The preparation and characterization of two triorganophosphinegold(I) complexes containing the anion derived from thiobenzoic acid are described. The cytotoxicity of these complexes has been investigated along with that of triphenylphosphinegold(I) mercaptopurinate, a known anti-tumor compound, against a variety of human cell lines. The complexes showed moderate to high cytotoxicity (ID(50) 250 - 2500 ng/ml).     

Chemoenzymatic synthesis of biotinylated nucleotide sugars as substrates for glycosyltransferases

The enzymatic oxidation of uridine 5'-diphospho-alpha-D-galactose (UDP-Gal) and uridine 5'-diphospho-N-acetyl-alpha-D-galactosamine (UDP-GalNAc) with galactose oxidase was combined with a chemical biotinylation step involving biotin-epsilon-amidocaproylhydrazide in a one-pot synthesis. The novel nucleotide sugar derivatives uridine 5'-diphospho-6-biotin-epsilon-amidocaproylhydrazino-alpha-D-galactose (UDP-6-biotinyl-Gal) and uridine 5'-diphospho-6-biotin-epsilon-amidocaproylhydrazino-N-acetyl-alpha-D-galactosamine (UDP-6-biotinyl-GalNAc) were synthesized on a 100-mg scale and characterized by mass spectrometry (fast atom bombardment and matrix-assisted laser desorption/ionization time of flight) and one/two dimensional NMR spectroscopy. It could be demonstrated for the first time, by use of UDP-6-biotinyl-Gal as a donor substrate, that the human recombinant galactosyltransferases beta3Gal-T5, beta4Gal-T1, and beta4Gal-T4 mediate biotinylation of the neoglycoconjugate bovine serum albumin-p-aminophenyl N-acetyl-beta-D-glucosaminide (BSA-(GlcNAc)17) and ovalbumin. The detection of the biotin tag transferred by beta3Gal-T5 onto BSA-(GlcNAc)17 with streptavidin-enzyme conjugates gave detection limits of 150 pmol of tagged GlcNAc in a Western blot analysis and 1 pmol of tagged GlcNAc in a microtiter plate assay. The degree of Gal-biotin tag transfer onto agalactosylated hybrid N-glycans present at the single glycosylation site of ovalbumin was dependent on the Gal-T used (either beta3Gal-T5, beta4Gal-T4, or beta4Gal-T1), which indicates that the acceptor specificity may direct the transfer of the Gal-biotin tag. The potential of this biotinylated UDP-Gal as a novel donor substrate for human galactosyltransferases lies in the targeting of distinct acceptor structures, for example, under-galactosylated glycoconjugates, which are related to diseases, or in the quality control of glycosylation of recombinant and native glycoproteins.     

Intuitive robust stability metric for PID control of self-regulating processes

Published methods establish how plant-model mismatch in the process gain and dead time impacts closed-loop stability. However, these methods assume no plant-model mismatch in the process time constant. The work presented here proposes the robust stability factor metric, RSF, to examine the effect of plant-model mismatch in the process gain, dead time, and time constant. The RSF is presented in two forms: an equation form and a visual form displayed on robustness plots derived from the Bode and Nyquist stability criteria. This understanding of robust stability is reinforced through visual examples of how closed-loop performance changes with various levels of plant-model mismatch. One example shows how plant-model mismatch in the time constant can impact closed-loop stability as much as plant-model mismatch in the gain and/or dead time. Theoretical discussion shows that the impact is greater for small dead time to time constant ratios. As the closed-loop time constant used in Internal Model Control (IMC) tuning decreases, the impact becomes significant for a larger range of dead time to time constant ratios. To complete the presentation, the RSF is used to compare the robust stability of IMC-PI tuning to other PI, PID, and PID with Filter tuning correlations.     

Shall we dance? How a multicopper oxidase chooses its electron transfer partner

Multicopper oxidases (MCOs) are encoded in the genomes of Eukarya, Bacteria, and Archea. These proteins are unique in that they contain at least four Cu atom prosthetic groups organized into one each of the three spectral classifications of copper sites in biology: type 1 (T1), type 2 (T2), and binuclear type 3 (T3), where the T2 and T3 sites form a trinuclear Cu cluster. With these four redox-active copper sites, the multicopper oxidases catalyze the four-electron (4e(-)) reduction of dioxygen to 2H2O, an activity that they alone share with the terminal heme-containing oxidases. Most MCOs exhibit broad specificity towards organic reductants, while a relatively small number of family members exhibit equally robust activity towards metal ions like Fe(II), Cu(I), and Mn(II) and, thus, are considered metallo-oxidases. This Account analyzes the structure-activity features of multicopper oxidases that determine their relative substrate specificity. Since the substrate oxidation step involves an outer-sphere electron transfer from the reductant to the T1Cu site in the protein, the concepts of Marcus theory are applied to unravel the origin of the substrate specificity of the multicopper ferroxidases.