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At least 811 records · Page 45Linked to original sources
PubMed2002

Analysis of biomolecular interactions using a miniaturized surface plasmon resonance sensor.

A commercially available miniaturized surface plasmon resonance sensor has been investigated for its applicability to biological interaction analysis. The sensor was found to exhibit excellent repeatability and linearity for high-refractive index solutions and good reproducibility for the binding of proteins. Its detection limit for the monoclonal antibody M1 was found to be 2.1 fmol, which corresponds to a surface concentration of 21 pg/mm2. Simple surface immobilization procedures relying on biotin/avidin or glycoprotein/lectin chemistry have been explored. Equilibrium dissociation constants for the binding of the FLAG peptide to its monoclonal antibody (M1) and for the binding of concanavalin A to a glycoprotein have been determined. The close agreement of these measurements with values obtained by surface fluorescence microscopy and fluorescence correlation spectroscopy helps to validate the use of this device. Thus, this sensor shows promise as an inexpensive, portable, and accurate tool for bioanalytical applications in laboratory and clinical settings.

Animals↗
PubMed2003

Miniaturized electrochemical flow cells.

Several novel types of miniaturized electrochemical flow cells are described. The flow cells are fabricated in fluorinated ethylene propylene using a novel technique where channels with inner diameters down to 13 microm are integrated with electrodes. The channel is formed by shrinking and simultaneous melting of a heat shrink/melt tubing around a channel template (a tungsten wire) and electrodes followed by removal of the channel template. The technique allows incorporation of different electrode materials of different sizes. The electrode configuration consists of one or two working electrodes inside the channel and a counter electrode located in the channel outlet reservoir. Electrode configurations with different channel and working electrode sizes, different electrode materials including carbon fibers, glassy carbon rods, poly(tetrafluoroethylene)/carbon composite material, and platinum wires, and different arrangements have been assembled. Hydrodynamic voltammograms in dual-electrode (generator-collector) experiments indicate good potential control for cells with 25-microm channels, while there is some iR drop in cells with 13-microm channels. Cells prepared with a cylindrical working electrode tangent and perpendicular to a flow channel show a flow rate dependence consistent with thin-layer cell behavior. Electrode areas can be made in the range of 10(-10)-10(-8) m2.

Ascorbic Acid↗
PubMed2003

Bacillus spore identification via proteolytic peptide mapping with a miniaturized MALDI TOF mass spectrometer.

An approach is tested here as a rapid screening method for Bacillus spore species employing bacterial peptide analysis with a miniaturized MALDI TOF mass spectrometer. A limited set of tryptic peptides was generated in situ following selective solubilization of the small, acid-soluble protein family (SASP) from spore samples on the MALDI sample holder. To facilitate species identification, a compact database was created comprising masses of the tryptic cleavage products generated in silico from all Bacillus and Clostridium SASPs whose sequences are available in public databases. Experimental measurements were matched against the custom-made database, and a published statistical model was then used to evaluate the probability of false identifications.

Bacillus↗
PubMed2004

Reactions and fluidics in miniaturized natural convection systems.

Buoyancy-driven convection offers a novel and greatly simplified mechanism for generating continuous nonpulsatile flow fields and performing thermally activated biochemical reactions. In this paper, we build on our previous work by constructing a multiwell device incorporating an array of 35-microL cylindrical cavities to perform polymerase chain reaction (PCR) amplification of a 191-base pair fragment associated with membrane channel proteins M1 and M2 of the influenza-A virus in as little as 15 min with performance comparable to conventional thermocyclers. We also describe entirely new adaptations of convective flows by conducting a series of coordinated flow visualization and computational studies to explore the design of closed-loop systems to execute tunable thermocycling, pumping, and mixing operations in a format suitable for integration into miniaturized biochemical analysis systems. Using 15-microL convective flow loops, we are able to perform PCR amplification of the same 191-base pair fragment associated with the influenza-A virus, as well as a 295-base pair segment of the human beta-actin gene in a format offering an enhanced degree of control and tunability. These convective flow devices can be further scaled down to nanoliter volumes and are ideally suited as a platform for a new generation of low-power, portable microfluidic DNA analysis systems.

Actins↗
PubMed2004

Miniaturization of frit inlet asymmetrical flow field-flow fractionation.

A miniaturized frit inlet asymmetrical flow field-flow fractionation (mFI-AFlFFF) channel has been constructed and tested for the separation of proteins. By scaling down the geometrical channel dimension of a conventional FI-AFlFFF system, flow rate ranges that can be manipulated were decreased to 20-30 microL/min, which reduces the injection amount of sample materials. The end effect contribution to plate height was evaluated by varying the inner diameter of the connection tubing between the injector and the channel inlet at various injection flow rates, and the results showed that the use of silica capillary tubing of the shortest possible distance is essential in reducing the initial band broadening prior to the sample injection to the microscale channel. The capability of the microFI-AFlFFF system was demonstrated with the separation of protein standards, polystyrenesulfonates, and ssDNA strains and for the characterization of replication protein A-ssDNA binding complex regulated by redox status.

Alcohol Dehydrogenase↗
PubMedDate not supplied

Constant-volume hydrogel osmometer: a new device concept for miniature biosensors.

A new type of biosensor is proposed that combines the recognition properties of "intelligent" hydrogels with the sensitivity and reliability of microfabricated pressure transducers. In the proposed device, analyte-induced changes in the osmotic swelling pressure of an environmentally responsive hydrogel are measured by confining it within a small implantable enclosure between a rigid semipermeable membrane and the diaphragm of a miniature pressure transducer. Proof-of-principle tests of this device were performed in vitro using pH-sensitive hydrogels, with osmotic deswelling data for the same hydrogels used as a benchmark for comparison. The swelling pressure of the hydrogel was accurately determined from osmotic deswelling measurements against reservoirs of known osmotic stress. Values of swelling pressure vs salt concentration measured with a preliminary version of the sensor agree well with osmotic deswelling results. Through modification of the hydrogel with various enzymes or pendant binding moieties, the sensor has the potential to detect a wide range of biological analytes with good specificity.

Biosensing Techniques↗
PubMed2006

High power density from a miniature microbial fuel cell using Shewanella oneidensis DSP10.

A miniature microbial fuel cell (mini-MFC) is described that demonstrates high output power per device cross-section (2.0 cm2) and volume (1.2 cm3). Shewanella oneidensis DSP10 in growth medium with lactate and buffered ferricyanide solutions were used as the anolyte and catholyte, respectively. Maximum power densities of 24 and 10 mW/m2 were measured using the true surface areas of reticulated vitreous carbon (RVC) and graphite felt (GF) electrodes without the addition of exogenous mediators in the anolyte. Current densities at maximum power were measured as 44 and 20 mA/m2 for RVC and GF, while short circuit current densities reached 32 mA/m2 for GF anodes and 100 mA/m2 for RVC. When the power density for GF was calculated using the cross sectional area of the device or the volume of the anode chamber, we found values (3 W/m2, 500 W/m3) similar to the maxima reported in the literature. The addition of electron mediators resulted in current and power increases of 30-100%. These power densities were surprisingly high considering a pure S. oneidensis culture was used. We found that the short diffusion lengths and high surface-area-to-chamber volume ratio utilized in the mini-MFC enhanced power density when compared to output from similar macroscopic MFCs.

Anthraquinones↗
PubMed1999

Lack of variation in alphaGal expression on lymphocytes in miniature swine of different genotypes.

BACKGROUND: Gal(alpha)1-3Gal epitopes (alphaGal) have been demonstrated to be present on tissues of all pig breeds tested to-date and are the major target for human anti-(alpha)galactosyl (alphaGal) antibodies. We investigated members of an MHC-inbred miniature swine herd to assess whether there was an association between genotype and expression of alphaGal. Identification of a low expressor genotype would potentially enable selective breeding of pigs that might prove beneficial as donors in clinical xenotransplantation. METHODS: we measured alphaGal expression on various pig cells by use of fluorescent-activated cell sorter (FACS) using (i) purified human anti-alphaGal antibody and (ii) the isolectin GS-I-B4. Initial studies were on porcine peripheral blood mononuclear cells (PBMCs) and subsequent studies on lymphocytes, platelets, and T cell subsets (CD4+ and CD8+ cells). RESULTS: there was considerable day-to-day variation in alphaGal expression on PBMCs from the same pig. When only lymphocytes were examined, there was a high degree of reproducibility, and no significant difference in alphaGal expression was detected between representative pairs of animlas of three different genotypes. Purified anti-alphaGal antibody bound to different sites on the alphaGal epitope than did Griffonia (Bandeiraea) simplicifolia I-B4 (GS-I-B4). Lectin binding was significantly reduced in the absence of divalent cations. When CD4+ and CD8+ T cells were examined for alphaGal expression, two distinct populations of each type of cell were observed, with larger cells expressing a higher level of alphaGal. CONCLUSIONS: although the number of pigs of different genotypes studied was small, on the basis of this limited study, pigs of a low alphaGal expressor genotype that could be selectively bred for use in clinical xenotransplantation were not identified.

Animals↗
PubMed1998

Evaluation of filling materials in membrane--protected bone defects. A comparative histomorphometric study in the mandible of miniature pigs.

In recent years, bone grafts and bone substitutes have been increasingly utilized underneath barrier membranes to optimize the treatment outcome of bone reconstructive therapy for defects in the alveolar process. In the present study, 4 different filling materials were evaluated in bone defects of similar dimensions in the mandible of miniature pigs. Blood clots and autografts were used as controls. The defects were covered with barrier membranes and allowed to heal for 4, 12 or 24 weeks. Histologic examination demonstrated that bone repair progressed through a programmed sequence of maturation steps closely resembling the pattern of bone development and growth regardless of whether bone grafts or substitutes were present or not. Histomorphometric analysis showed that autologous bone grafts (autografts) had the best osteoconductive properties during the initial healing period, with 39% of newly formed bone inside the membrane-covered defects at 4 weeks of healing. In addition, 87% of the graft surfaces were already covered by bone at this time. Both values were significantly higher for autografts than for the 4 alternative bone fillers (P < or = 0.05). At 12 weeks, these differences were no longer apparent, with all 5 filling materials showing similar values. Among the tested bone substitutes, tricalcium phosphate (TCP) showed a significantly higher percentage of bone fill at 24 weeks of healing. It can be concluded that sites filled with autografts clearly demonstrated the best results underneath barrier membranes in the early phase of healing. As far as degradation and substitution are concerned, TCP showed the most promising results. This filler, however, needs to be tested further in a more demanding animal model. Less favorable results were obtained for coral-derived hydroxyapatite granules and for demineralized freeze-dried bone allografts.

Alveolar Bone Loss↗
PubMed1999

Sinus floor augmentation with simultaneous placement of dental implants using a combination of deproteinized bone xenografts and recombinant human osteogenic protein-1. A histometric study in miniature pigs.

Maxillary sinus floor augmentation with autogenous bone has become a widely accepted procedure in implant dentistry. The use of osteoconductive bone substitutes in this indication is controversial, since their use can lead to a prolonged healing time, inhomogenous ossification, foreign body reaction, migration of particles and low bone-implant contact (BIC). The purpose of this study was to examine whether the combination of an osteoinductive protein (recombinant human osteogenic protein-1 (rhOP-1 = bone morphogenetic protein-7) with natural bovine bone mineral (BioOss) would improve ossification and the bone-implant contact (BIC) in a sinus floor augmentation with simultaneous placement of implants. In this study, the maxillary sinus floors in 5 miniature pigs were augmented with 3 ml BioOss containing 420 micrograms rhOP-1 on the test side and 3 ml BioOss alone on the control side. At the time of augmentation a titanium implant (ITI) was inserted from a laterocaudal direction. After 6 months of healing the sites of augmentation were removed and examined in non-decalcified sections by microradiography, fluorescence microscopy of sequentially labelled specimens and by histometry. On both sides, significant amounts of newly-formed bone were observed. However, on the test sites, the percentage of BIC in the augmented area was 80.0% versus 38.6% on control sites. It can be concluded that the application of bone morphogenetic proteins caused a more rapid and enhanced osseointegration of simultaneously placed implants when compared to the bone substitute alone. Therefore recombinant human osteogenic protein-1 delivered by natural bone mineral has the potential to become a clinical alternative for autogenous bone grafts in sinus floor augmentation.

Animals↗
PubMed2005

Miniaturization in functional genomics and proteomics.

Proteins are the key components of the cellular machinery responsible for processing changes that are ordered by genomic information. Analysis of most human proteins and nucleic acids is important in order to decode the complex networks that are likely to underlie many common diseases. Significant improvements in current technology are also required to dissect the regulatory processes in high-throughtput and with low cost. Miniaturization of biological assays is an important prerequisite to achieve these goals in the near future.

Biological Assay↗
PubMed1998

Sialographic characterization of the normal parotid gland of the miniature pig.

OBJECTIVE: To characterize the structure of the parotid gland of the miniature pigs (minipig). METHODS: Sialographic, anatomical, histological and ultrastructural studies of the parotid gland were performed on 11 minipigs. RESULTS: Sialograms showed a long main duct and a triangular shaped gland. All branching ducts extended from the inferior-posterior margin of the main duct. No accessory glands were found. Typical serous acini were found microscopically and histochemically. CONCLUSION: This study provides basic structural information on the parotid gland of the minipig.

Animals↗
PubMed2001

Miniaturized flow-through PCR with different template types in a silicon chip thermocycler.

Flow-through chip thermocyclers can be used in miniaturized rapid polymerase chain reaction (PCR) despite their high surface to volume ratio of samples. We demonstrated that a thermocycler made of silicon and glass chips and containing thin film transducers for heating and temperature control can be adapted to the amplification of various DNA templates of different sources and properties. Therefore, the concept of serial flow in a liquid/liquid two-phase system was combined with a surface management of inner side walls of the microchannel and an adaptation of PCR mixture composition. In addition, the process temperatures and the flow rates were optimized. Thus, a synthetic template originating from investigations on nucleic acid evolution with 106 base pairs [cooperative amplification of templates by cross hybridization (CATCH)], a house keeping gene with 379 base pairs [glutaraldehyde 3-phosphate dehydrogenase (GAPDH)] and a zinc finger protein relevant in human pathogenesis with 700 base pairs [Myc-interacting zinc finger protein-1, knock-out (Miz1-KO)] were amplified successfully. In all three cases the selectivity of priming and amplification could be shown by gel electrophoresis. The typical amplification time was 1 min per temperature cycle. So, the typical residence time of a sample volume inside the 25 cycle device amounts to less then half an hour. The energy consumption of the PCR chip for a 35 min PCR process amounts to less than 0.012 kW h.

Base Sequence↗
PubMed1999

Influence of platelet activating factor on gastrointestinal electrical activity and some haematological and clinical parameters in the conscious miniature pig.

The effects of intravenous (i.v.) infusion of platelet-activating factor (PAF), 100 ng/kg/min for 10 min, with and without pretreatment with a selective PAF-antagonist on gastrointestinal electrical activity, arterial pressure and clinical and haematological parameters were studied. Conscious miniature pigs with electrodes implanted in the wall of the antrum pylori and small and large intestine were used. Platelet-activating factor induced restlessness or depression, shivering, tachypnoea and coughing, retching and vomiting, hypotension and a delayed and sustained increase in leucocyte count with an increase in percentage of segmented neutrophils. The PAF-antagonist, SAH 63-675, administered at 10 mg/kg intravenously, inhibited these effects. Platelet-activating factor resulted in a decrease in electrical activity in the antrum and large intestine, whereas small intestinal activity was not significantly influenced. Pretreatment with the antagonist suppressed these inhibitory effects.

Animals↗
PubMed2000

Involvement of platelet activating factor in the endotoxin-induced effects on gastrointestinal electrical activity and some haematological parameters in the conscious miniature pig.

In conscious miniature pigs the influence of intravenous dose of lipopolysaccharide (LPS), 10 microg/kg over 10 min, with and without pretreatment with a platelet activating factor (PAF) antagonist, SAH 63-675 10 mg/kg, on gastrointestinal electrical activity, arterial pressure and clinical and haematological parameters was studied. Dose of LPS provoked mild clinical signs and hypotension, which were prevented by PAF antagonism. The LPS induced leukocytosis and increase in mature neutrophils, however, were PAF independent. Pretreatment with the PAF antagonist attenuated the LPS-provoked inhibition of electrical activity in the antrum, jejunum, ileum and caecum. These results suggest a beneficial effect of PAF antagonism in porcine endotoxaemia.

Animals↗
PubMedDate not supplied

Dermal regeneration in full-thickness wounds in Yucatan miniature pigs using a biodegradable copolymer.

The aim of this study was to assess the performance of a biodegradable dermal substrate in deep dermal skin defects. The substrate consisted of a synthetic biodegradable matrix called Polyactive, which is an elastomeric poly (ether)/ poly (ester) block copolymer. This matrix was manufactured either as a porous substrate, with gradually changing pore size (BISKIN-M), or as a bilayer consisting of a porous underlayer with a fully dense surface layer (BISKIN). Cell-free matrices and matrices seeded with autologous or allogeneic porcine fibroblasts were applied to full-thickness skin wounds in Yucatan miniature pigs. Biopsies were taken at different time intervals up to 24-months post-transplantation. Although all BISKIN substrates showed little or no adherence to the wound bed, the adherence of the BISKIN-M substrates to the underlying wound was achieved within minutes after application. Therefore, only BISKIN-M Polyactive substrates were further evaluated. Wound contraction was inhibited by both cell-free and fibroblast-populated substrates. All substrates showed extensive neovascular and fibrous tissue ingrowth within 2-weeks post-transplantation. Furthermore, during this time period, matrix degradation was observed, starting with the fragmentation of the polymers into particles, which were phagocytized by macrophages. These processes occurred actively up to 3 months and ceased thereafter. Cell-free substrates degraded faster, and also, the collagen deposition was lower as compared with cell-seeded substrates. The tissue surrounding the remnants of the Polyactive substrates after 24-months post-transplantation consisted of a mature connective tissue. The newly formed collagen had the same distribution pattern as observed in normal native dermis. We conclude therefore that treatment of full-thickness skin defects with fibroblast-populated BISKIN-M Polyactive substrates leads to satisfactory dermal regeneration.

Absorbable Implants↗
PubMed2000

Impeller design for a miniaturized centrifugal blood pump.

The impeller design for a miniature centrifugal blood pump is an important consideration since the small diameter impeller requires higher rotational speed, which may cause more blood trauma compared to the larger diameter impeller. Three different impeller vanes (straight vanes with a height of 4 mm and 8 mm, and 8 mm curved vanes) of which the diameter was 35 mm were subjected to hydraulic performance and hemolysis tests in the same pump housing. Both straight vane impellers attained left ventricular assist condition (5 L/min against 100 mm Hg) at 2,900 rpm while the curved vane required 3,280 rpm. There was no significant hemolysis difference between the tall and short vanes. The curved impeller vanes did not exhibit sufficient hydraulic performance when compared to the straight vanes. The straight vane impellers, even with different heights, were incorporated into the same pump housings, and the vane heights did not drastically change the hydraulic performance or hemolysis.

Equipment Design↗