Seventeen-year survivorship analysis of silastic metacarpophalangeal joint replacement .
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We reviewed the records and radiographs of 381 patients with rheumatoid arthritis who had undergone silastic metacarpophalangeal joint replacement during the past 17 years. The number of implants was 1336 in the course of 404 operations. Implant failure was defined as either revision or fracture of the implant as seen on radiography. At 17 years, the survivorship was 63%, although on radiographs two-thirds of the implants were seen to be broken. Factors which improved survival included soft-tissue balancing, crossed intrinsic transfer and realignment of the wrist. Surgery to the thumb and proximal interphalangeal joint had a deleterious effect and the use of grommets did not protect the implant from fracture. (+info)
A picoliter chamber array for cell-free protein synthesis.
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The completion of human genome sequencing has shifted the focus of research from genes to proteins. In this regard, a protein library chip has become a useful tool for cell-free protein synthesis. In this study, we attempted to make a highly-integrated protein chip from a DNA library using in vitro protein synthesis on a microchamber array fabricated by using PDMS (polydimethyl siloxane), a hydrophobic surface, and glass, a hydrophilic bottom substrate. These structural properties prevented cross-contamination among the chambers. The minimum volume capacity of the smallest chamber was about 1 pl. The total number of chambers per chip was 10,000 on one chip (capacity 150 pl) and 250,000 on two others (1 and 5 pl). Next, we attempted in vitro protein synthesis using this microchamber array. The fluorescence of Green Fluorescent Protein (GFP) expressed on the chamber was rapidly detected (within just 1 h). GFP expression was also successful using immobilized DNA molecules on polymer beads. DNA immobilized beads were added as the source to each microchamber. Protein was successfully synthesized from DNA immobilized beads, which allowed easy handling of the DNA molecules. (+info)
Retinal pigment epithelial cell behavior is modulated by alterations in focal cell-substrate contacts.
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PURPOSE: To investigate how the cellular behavior of cultured retinal pigment epithelial (RPE) cells was affected by the manipulation of early focal contact. METHODS: To manipulate early focal contact, a reduced focal cell-substrate contact area on the micropatterned surfaces was implemented by microfabrication with polydimethylsiloxane (PDMS). The micropatterned PDMS surfaces had a circular pillar with a diameter of 5 microm. The human retinal pigment epithelial cell line, ARPE-19, was seeded onto the fibronectin-coated PDMS surfaces. Cell adhesion, growth, cell cycle, morphology, and interleukin-6 (IL-6) expression were observed for 3 weeks. RESULTS: The fluorescent images of localized vinculin and actin stress fibers appeared to be more prominent on smooth PDMS surfaces. Although there was no significant effect on cell adhesion, a statistically significant inhibition of cell cycle progression was observed for micropatterned PDMS surfaces. Similarly, micropatterned surfaces showed significantly less cell growth than that of smooth surfaces. Cultures over a period of 3 weeks showed a distinct cell-cell phenotype discrepancy. Furthermore, IL-6 mRNA and secreted protein induced by IL-1beta in ARPE-19 were downregulated on micropatterned PDMS surfaces. CONCLUSIONS: Disturbed focal contact in ARPE-19 cells grown on micropatterned surfaces altered cell cycle, growth, morphology, and the expression of IL-6 in vitro. (+info)
Controlling nonspecific protein adsorption in a plug-based microfluidic system by controlling interfacial chemistry using fluorous-phase surfactants.
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Control of surface chemistry and protein adsorption is important for using microfluidic devices for biochemical analysis and high-throughput screening assays. This paper describes the control of protein adsorption at the liquid-liquid interface in a plug-based microfluidic system. The microfluidic system uses multiphase flows of immiscible fluorous and aqueous fluids to form plugs, which are aqueous droplets that are completely surrounded by fluorocarbon oil and do not come into direct contact with the hydrophobic surface of the microchannel. Protein adsorption at the aqueous-fluorous interface was controlled by using surfactants that were soluble in fluorocarbon oil but insoluble in aqueous solutions. Three perfluorinated alkane surfactants capped with different functional groups were used: a carboxylic acid, an alcohol, and a triethylene glycol group that was synthesized from commercially available materials. Using complementary methods of analysis, adsorption was characterized for several proteins (bovine serum albumin (BSA) and fibrinogen), including enzymes (ribonuclease A (RNase A) and alkaline phosphatase). These complementary methods involved characterizing adsorption in microliter-sized droplets by drop tensiometry and in nanoliter plugs by fluorescence microscopy and kinetic measurements of enzyme catalysis. The oligoethylene glycol-capped surfactant prevented protein adsorption in all cases. Adsorption of proteins to the carboxylic acid-capped surfactant in nanoliter plugs could be described by using the Langmuir model and tensiometry results for microliter drops. The microfluidic system was fabricated using rapid prototyping in poly(dimethylsiloxane) (PDMS). Black PDMS microfluidic devices, fabricated by curing a suspension of charcoal in PDMS, were used to measure the changes in fluorescence intensity more sensitively. This system will be useful for microfluidic bioassays, enzymatic kinetics, and protein crystallization, because it does not require surface modification during fabrication to control surface chemistry and protein adsorption. (+info)
BACKGROUND: Bone marrow-derived cell populations possess progenitor cell capacities. Emerging evidence also suggests significant plasticity of differentiated mononuclear cell lineages. We therefore assessed the distribution of transplanted peripheral blood mononuclear cells (PBMCs) in granulation tissue formation, and evaluated their possible transdifferentiation into myofibroblasts. METHODS: Silastic tubes were inserted into the peritoneal cavity of rats, followed by injection of PKH26-labelled PBMCs isolated from donor animals. At 3, 14 and 21 days, the distribution of PKH26(+) cells as well as their colocalization with myofibroblast/smooth muscle cell [alpha-smooth muscle (alpha-SM) actin] or macrophage markers (ED1/ED2) were determined. RESULTS: Round-shaped PKH26(+) cells accumulated around the implants at 3 days, while myofibroblasts were rare. Later, peritoneal granulation tissue constituted an inner, multilayered capsule primarily comprising alpha-SM actin(+) cells that was surrounded by more loosely organized inflammatory connective tissue. PKH26-labelled, spindle-shaped cells were abundantly found in tissue capsules. As a key finding, granulation tissue at 14 and 21 days contained cells with both PKH26 and alpha-SM actin labelling. Accordingly, a subpopulation of cells staining positive for macrophage markers showed a spindle-shaped morphology and alpha-SM actin expression. CONCLUSIONS: Transplanted PBMCs contribute to granulation tissue, and acquire myofibroblast characteristics during de novo tissue formation. Mononuclear cells may transdifferentiate into myofibroblast-like cells within an inflammatory environment. (+info)
Androgen control of immunocompetence in the male house finch, Carpodacus mexicanus Muller.
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The immunocompetence handicap (ICH) hypothesis predicts that elevated levels of the gonadal androgen testosterone (T) entail obligatory costs, such as immunosuppression, but evidence supporting this immunosuppressive influence is equivocal. To investigate this question, adult males house finches, Carpodacus mexicanus, were exposed to short days and chronically treated with T-filled (T males; N=10) or empty (C males; N=10) Silastic capsules. Testosterone administration increased plasma T levels and the size of the cloacal protuberance, an androgen-dependent secondary sexual characteristic. To study humoral immunity, finches received injections of sheep red blood cells (SRBC) and we measured circulating concentrations of antibodies to these cells with a hemagglutination test. All males produced antibodies following four SRBC injections at weekly intervals. Antibody titers in T and C males did not differ 5 days after the fourth injection, but were 59% lower in T than C males 2 weeks later. To study cell-mediated immunity, we measured the local inflammatory response to an injection of phytohemaglutinin (PHA). This response in T and C males was similar 1 day after PHA injection, but was 58% less in T than C males 2 days following the injection. Thus, T and C males mounted similar humoral and cell-mediated immune responses, but T treatment compromised maintenance of these responses. The results, demonstrating immunosuppressive effects of elevated T, are consistent with the ICH hypothesis. (+info)
In situ oligonucleotide synthesis on poly(dimethylsiloxane): a flexible substrate for microarray fabrication.
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In this paper, we demonstrate in situ synthesis of oligonucleotide probes on poly(dimethylsiloxane) (PDMS) microchannels through use of conventional phosphoramidite chemistry. PDMS polymer was moulded into a series of microchannels using standard soft lithography (micro-moulding), with dimensions <100 microm. The surface of the PDMS was derivatized by exposure to ultraviolet/ozone followed by vapour phase deposition of glycidoxypropyltrimethoxysilane and reaction with poly(ethylene glycol) spacer, resulting in a reactive surface for oligonucleotide coupling. High, reproducible yields were achieved for both 6mer and 21mer probes as assessed by hybridization to fluorescent oligonucleotides. Oligonucleotide surface density was comparable with that obtained on glass substrates. These results suggest PDMS as a stable and flexible alternative to glass as a suitable substrate in the fabrication and synthesis of DNA microarrays. (+info)
Treatment of head louse infestation with 4% dimeticone lotion: randomised controlled equivalence trial.
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OBJECTIVE: To evaluate the efficacy and safety of 4% dimeticone lotion for treatment of head louse infestation. DESIGN: Randomised controlled equivalence trial. SETTING: Community, with home visits. PARTICIPANTS: 214 young people aged 4 to 18 years and 39 adults with active head louse infestation. INTERVENTIONS: Two applications seven days apart of either 4.0% dimeticone lotion, applied for eight hours or overnight, or 0.5% phenothrin liquid, applied for 12 hours or overnight. OUTCOME MEASURES: Cure of infestation (no evidence of head lice after second treatment) or reinfestation after cure. RESULTS: Cure or reinfestation after cure occurred in 89 of 127 (70%) participants treated with dimeticone and 94 of 125 (75%) treated with phenothrin (difference -5%, 95% confidence interval -16% to 6%). Per protocol analysis showed that 84 of 121 (69%) participants were cured with dimeticone and 90 of 116 (78%) were cured with phenothrin. Irritant reactions occurred significantly less with dimeticone (3/127, 2%) than with phenothrin (11/125, 9%; difference -6%, -12% to -1%). Per protocol this was 3 of 121 (3%) participants treated with dimeticone and 10 of 116 (9%) treated with phenothrin (difference -6%, -12% to -0.3%). CONCLUSION: Dimeticone lotion cures head louse infestation. Dimeticone seems less irritant than existing treatments and has a physical action on lice that should not be affected by resistance to neurotoxic insecticides. (+info)