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PubMed1975

Model system for studying colonization and growth of bacteria on a hydroxyapatite surface.

A model system for the study of bacterial colonization and growth on a hydroxyapatite (HT) surface is described. Hydroxyapatite was crystallized over the surface of porous glass beads. Chemical analysis of the product showed that the ratio of Ca2+/P042- was indistinguishable from that of commercial HT powder. X-ray diffraction analysis supported the conclusion that the product was HT. A system employing [14C]polyethylene glycol, which selectively adsorbs to the glass surface of the beads, was developed to determine the amount of glass surface covered by HT. Over 90% of the glass surface could be covered by our method. The product, HT beads, consisted of approximately 20% (dry weight) HT. The HT beads possess several properties which make them potentially useful for studying microbial adherence, growth, and interactions. These include: (i) chemical similarity to the tooth surface, (ii) large surface area, and (iii) high density. We also describe a method for direct measurement of the microbial mass of cells growing on beads. The method entails immobilizing a sample on a membrane filter (Millipore), staining it with amido black dye, and eluting the dye for spectrophotometric measurement. Streptococcus mutans served as the test organism. For free-growing bacteria the values measured with the filter assay were directly proportional to cell number, with a value of 1 mug of "protein" corresponding to about 1.5 X 10(6) colony-forming units, determined by viable count. For bacteria colonizing the beads, 1 mug of protein corresponded to about 2 X 10(7) colony-forming units on the beads during logarithmic growth. As the culture approached stationary phase, the efficiency of the assay decreased. These data indicate that multiple random samples, taken at a given time, are representative of the entire culture.

Bacteriological Techniques↗
PubMed2001

Yeast as a model system for anticancer drug discovery.

Saccharomyces cerevisiae has been used extensively as a model for higher eukaryotes in the study of basic cellular processes. The high degree of conservation in terms of sequence similarity and function has made this organism useful in elucidating biological pathways, both yeast and human. Among these are pathways responsible for DNA damage repair and cell cycle control. This review presents an overview of opportunities for using yeast as a model system for anticancer drug discovery. It covers screens directed against specific cancer-related targets as well as contexts created by cancer-related alterations. The methodologies covered include pharmacological and genetic screens, as well as genome-wide approaches to drug target identification.

Journal Article↗
PubMed1975

Murexide for determination of free and protein-bound calcium in model systems.

The determination with murexide of free and protein-bound calcium in model systems of known composition, ionic strength, and pH was investigated. The spectra of calcium murexide in the presence of varying amounts of calcium ions indicated that the absorption maximum fo calcium murexide complex occurs at 480 nm while that of murexide ion is at 520 nm. The absorbance at 509 nm is independent of calcium ion concentration and, therefore, could be used to measure the total dye. The spectra are pH dependent but constant in the range 6.5 to 7.0. The apparent dissociation constant of calcium murexide is dependent upon ionic environment, ionic strength, and free calcium ion concentration. The relationship between the apparent dissociation constant and free calcium concentration was established. Whole casein had no effect on the absorption spectra of calcium murexide and no affinity for calcium murexide complex or murexide ion. Beta-casein, at the concentrations employed, did not influence the dissociation fo calcium murexide. At pH 7.0, ionic strength .1, and 2 C, Beta-casein bound calcium as if there were 8.65 binding sites per molecule, each of pK 2.23, corresponding to an intrinsic association constant of 168.9 liters per mole.

Barbiturates↗
PubMed1998

High-pressure-mediated dissociation of immune complexes demonstrated in model systems.

The use of pressure to disrupt immune complexes was demonstrated in two model systems: prostate-specific antigen (PSA) and anti-PSA antibody; and epiglycanin, a mucin glycoprotein, and an antibody specific to that protein. Dissociation of the anti-PSA antibody from the immobilized PSA antigen was observed when pressures of 415 MPa and 550 MPa (1 MPa approximately 144 psi) were applied at room temperature (approximately 21 degrees C). Application of pressures ranging from 140 MPa to 550 MPa resulted in dissociation of antibody from epiglycanin. In both cases, the rebinding of dissociated antibody to immobilized antigen indicated that the effect of high pressure on the binding of the immune complexes was reversible. These findings suggest that application of high hydrostatic pressure has the potential to be used to significantly improve the sensitivity and specificity of clinical assays.

Antibodies↗
PubMed2004

Parallel processing in an identified neural circuit: the Aplysia californica gill-withdrawal response model system.

The response of the gill of Aplysia calfornica Cooper to weak to moderate tactile stimulation of the siphon, the gill-withdrawal response or GWR, has been an important model system for work aimed at understanding the relationship between neural plasticity and simple forms of non-associative and associative learning. Interest in the GWR has been based largely on the hypothesis that the response could be explained adequately by parallel monosynaptic reflex arcs between six parietovisceral ganglion (PVG) gill motor neurons (GMNs) and a cluster of sensory neurons termed the LE cluster. This hypothesis, the Kupfermann-Kandel model, made clear, falsifiable predictions that have stimulated experimental work for many years. Here, we review tests of three predictions of the Kupfermann-Kandel model: (1) that the GWR is a simple, reflexive behaviour graded with stimulus intensity; (2) that central nervous system (CNS) pathways are necessary and sufficient for the GWR; and (3) that activity in six identified GMNs is sufficient to account for the GWR. The available data suggest that (1) a variety of action patterns occur in the context of the GWR; (2) the PVG is not necessary and the diffuse peripheral nervous system (PNS) is sufficient to mediate these action patterns; and (3) the role of any individual GMN in the behaviour varies. Both the control of gill-withdrawal responses, and plasticity in these responses, are broadly distributed across both PNS and CNS pathways. The Kupfermann-Kandel model is inconsistent with the available data and therefore stands rejected. There is, no known causal connection or correlation between the observed plasticity at the identified synapses in this system and behavioural changes during non-associative and associative learning paradigms. Critical examination of these well-studied central pathways suggests that they represent a 'wetware' neural network, architecturally similar to the neural network models of the widely used 'Perceptron' and/or 'Back-propagation' type. Such models may offer a more biologically realistic representation of nervous system organisation than has been thought. In this model, the six parallel GMNs of the CNS correspond to a hidden layer within one module of the gill-control system. That is, the gill-control system appears to be organised as a distributed system with several parallel modules, some of which are neural networks in their own right. A new model is presented here which predicts that the six GMNs serve as components of a 'push-pull' gain control system, along with known but largely unidentified inhibitory motor neurons from the PVG. This 'push-pull' gain control system sets the responsiveness of the peripheral gill motor system. Neither causal nor correlational links between specific forms of neural plasticity and behavioural plasticity have been demonstrated in the GWR model system. However, the GWR model system does provide an opportunity to observe and describe directly the physiological and biochemical mechanisms of distributed representation and parallel processing in a largely identifiable 'wetware' neural network.

Animals↗
PubMed2002

Zebrafish as an immunological model system.

Two decades of research have established the zebrafish (Danio rerio) as a significant model system for studying vertebrate development and gene structure-function relationships. Recent advances in mutation screening, the creation of genomic resources, including the Zebrafish Genome Project and the development of efficient transgenesis procedures, make this model increasingly attractive for immunological study.

Animals↗
PubMed2005

Exciplex fluorescence emission from simple organic intramolecular constructs in non-polar and highly polar media as model systems for DNA-assembled exciplex detectors.

Organic intramolecular exciplexes, N-(4-dimethylaminobenzyl)-N-(1-pyrenemethyl)amine (1) and N'-4-dimethylaminonaphthyl-N-(1-pyrenemethyl)amine (2), were used as model systems to reveal major factors affecting their exciplex fluorescence, and thus lay the basis for developing emissive target-assembled exciplexes for DNA-mounted systems in solution. These models with an aromatic pyrenyl hydrocarbon moiety as an electron acceptor appropriately connected to an aromatic dimethylamino electron donor component (N,N-dimethylaminophenyl or N,N-dimethylaminonaphthyl) showed strong intramolecular exciplex emission in both non-polar and highly polar solvents. The effect of dielectric constant on the maximum wavelength for exciplex emission was studied, and emission was observed for 1 and 2 over the full range of solvent from non-polar hydrocarbons up to N-methylformamide with a dielectric constant of 182. Quantum yields were determined for these intramolecular exciplexes in a range of solvents relative to that for Hoechst 33,258. Conformational analysis of 1 was performed both computationally and via qualitative 2D NMR using (1)H-NOESY experiments. The results obtained indicated the contribution of pre-folded conformation(s) to the ground state of 1 conducive to exciplex emission. This research provides the initial background for design of self-assembled, DNA-mounted exciplexes and underpins further development of exciplex-based hybridisation bioassays.

DNA↗
PubMed2000

Zebrafish: a model system for the study of human disease.

The zebrafish (Danio rerio) is a powerful model organism for the study of vertebrate biology, being well suited to both developmental and genetic analysis. Large-scale genetic screens have identified hundreds of mutant phenotypes, many of which resemble human clinical disorders. The creation of critical genetic reagents, coupled with the rapid progress of the zebrafish genome initiative directed by the National Institutes of Health, are bringing this model system to its full potential for the study of vertebrate biology, physiology and human disease.

Animals↗
PubMed1978

A teratoma model system.

The technique of genital ridge transplantation in mice was described. The use of this model system for the study of teratomas is stressed.

Animals↗
PubMedDate not supplied

Early morphological changes of vessels in an experimental model system.

The theories and models concerning the pathology of arteriosclerosis are briefly reviewed. The Lipofundin arteriosclerosis model developed in our laboratory is present together with the mechanism of development of sclerotic vascular lesions and the primary role of permeability changes in the whole process. The process of development of the fibrotic proliferative lesions induced by Lipofundin was visualized by a colloidal iron tracer for enhanced permeability and the consequent smooth muscle cell proliferation was also registered. The presence and importance of adventitial lymphatic drainage was observed in connection with the enhancement of permeability. The adherence of macrophages to the endothelium was noted. We think that the enhanced permeability results in the functional impairment of the endothelium and causes macrophages to adhere to the damaged sites. Macrophages produce growth factors which together with enhanced permeability induce smooth muscle cell proliferation leading ultimately to the arteriosclerotic changes of the vessel wall. The pathological process was inhibited or at least attenuated by a prostacyclin-analog (Iloprost, 6-alpha-carbacyclin, Schering AG). The action of the drug seems to consist of a direct reduction of proliferation, of maintenance of normal permeability of normal endothelial functions. The latter acts against the adherence of macrophages thus cancels the possible stimulation of smooth muscle cell proliferation by macrophage growth factors.

Animals↗
PubMed1995

Analysis of the inherent error of the TMS-1 Topographic Modeling System in mapping a radially aspheric surface.

The potential spherical bias of the Topographic Modeling System (TMS) in measuring a radially aspheric surface was investigated. The radii of curvature of an ellipsoidal surface were measured experimentally using the TMS, and calculated using both an instantaneous radius of curvature formula and a spherically biased algorithm for comparison. Measurements were made aligned and under four conditions of misalignment. A sphere with the same radius of curvature as the apex of the ellipsoid was used to document the fundamental performance of the system. The TMS demonstrated increasing error from center to periphery in measurement of an ellipsoid with 7.5 mm apical radius of curvature and 0.5 eccentricity. Greater than 3 D of error was present at 4 mm from the apex. The misalignment error was small compared to the inherent error in the system performance. The TMS does not accurately measure the instantaneous radii of curvature of an ellipsoidal surface due to an inherent spherical bias. In addition, the spherically biased system may not detect subtle shifts in curvature in the peripheral regions, causing potentially inaccurate reconstruction of patterns of curvature.

Algorithms↗
PubMed1995

Inverse expressions of the N-myc oncogene and beta 1 integrin in human neuroblastoma: relationships to disease progression in a nude mouse model system.

A nude mouse model for human neuroblastoma has been developed to examine possible relationships between amplification/over-expression of the N-myc oncogene and altered regulation of expression of specific integrin subunits during tumor progression. Subcutaneous (ectopic) or intra-adrenal (orthotopic) injection of the neuroblastoma cell lines SK-N-SH or IMR-32 has generated a number of derivative tumor cell lines. Tumor cell lines derived from SK-N-SH cells (which do not express N-myc) or IMR-32 cells (which over-express N-myc) produce tumors at higher rates when re-injected into the subcutaneous space of nude mice. Moreover, cell lines derived from tumors initiated by IMR-32 cells exhibit shorter latent periods than do IMR-32 cells direct from tissue culture. With regard to integrin subunit expression, SK-N-SH and related cell lines express high levels of beta 1 integrin, which is associated with the alpha 2 and alpha 3 integrin subunits (predominantly alpha 3). IMR-32 cells display reduced beta 1 expression, and that which is produced is not associated with common alpha subunits. LaN1 cells, which express N-myc at even higher levels than do IMR-32 cells, express even less beta 1. Interestingly, the tumor-derived cell lines (especially those from tumors initiated in adrenal glands) also exhibit reduced integrin expression compared with the parental cell lines; this reduction is associated with the enhanced tumor take rate observed when the cells are re-injected into nude mice. Our results raise the possibility of a relationship between over-expression of N-myc and down-regulation of beta 1 integrin expression (possibly some alpha subunits also). In addition, the data suggest that human neuroblastoma-derived cell lines which exhibit reduced integrin expression display more aggressive tumor growth in nude mice.

Animals↗
PubMed1984

Abnormal discharges and chaos in a neuronal model system.

Using the mathematical model of the pacemaker neuron formulated by Chay, we have investigated the conditions in which a neuron can generate chaotic signals in response to variation in temperature, ionic compositions, chemicals, and the strength of applied depolarizing current.

Animals↗
PubMed1990

On the chemical nature of DNA and RNA modification by a hemin model system.

In order to model the interaction of hemin with DNA and other polynucleotides, we have studied the degradation of DNA, RNA, and polynucleotides of defined structure by [meso-tetrakis(N-methyl-4-pyridyl)porphinato]manganese(III) (MnTMPP) + KHSO5. The activated porphyrin was shown to release adenine, thymine, and cytosine from DNA; RNA degradation afforded adenine, uracil, and cytosine. The same products were obtained from single- and double-stranded DNA oligonucleotides of defined sequence, and also from single-stranded DNA and RNA homopolymers. The overall yield of bases from the dode-canucleotide d(CGCT3A3GCG) was equal to 14% of the nucleotides present initially, indicating that each porphyrin catalyzed the release of approximately 4 bases. Although no guanine was detected as a product from any of the substrates studied, the ability of MnTMPP + KHSO5 to degrade guanine nucleotides was verified by the destruction of pGp, and by the appearance of bands corresponding to guanosine cleavage following treatment of 32P end labeled DNA restriction fragments with activated MnTMPP. Inspection of a number of sites of MnTMPP-promoted cleavage indicated that the process was sequence-selective, occurring primarily at G residues that were part of 5'-TG-3' or 5'-AG-3' sequences, or at T residues. Also formed in much greater abundance were alkali-labile lesions; these were formed largely at guanosine residues. Also studied was the degradation of a 47-nucleotide RNA molecule containing two hairpins. Degradation of the 5'-32P end labeled RNA substrate afforded no distinct, individual bands, suggesting that multiple modes of degradation may be operative.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed2002

Atropselective macrocyclization of diaryl ether ring systems: application to the synthesis of vancomycin model systems.

Vancomycin is the last line of defense available in the clinic for treating multidrug-resistant bacterial infections. Vancomycin contains two 16-membered diaryl ether macrocycles, each of which contains a stereogenic axis across the diaryl ether linkage. Since an effective total synthesis of vancomycin requires that these stereogenic axes be formed in a stereoselective manner, we have developed an atropselective variation of the triazene mediated diaryl ether forming reaction. This variation introduced an energetic penalty into the transition state of the undesired atropisomer. This reaction is used to synthesize the C-O-D diaryl ether macrocycle found in vancomycin with high diastereoselectivity (de > 90%), providing the naturally occurring atropisomeric configuration.

Anti-Bacterial Agents↗
PubMed1986

Cellular proliferation in the vitreous: the use of vitreous explants as a model system.

An in vitro model of cellular proliferation in the vitreous has been developed using explants of bovine vitreous gel. Various cell types, including chick embryo pigmented retinal epithelium, choroidal fibroblasts, retinal glial cells, bovine retinal capillary endothelial cells, and peritoneal mouse macrophages, were cultured at 37 degrees C on vitreous gels for periods up to 8 days and compared for their effects on the structure of the gel. Choroidal fibroblasts caused a very marked reduction in the size of the gel and produced the strongest traction on the gel fibril structure; in contrast, peritoneal macrophages caused virtually no reduction in vitreous volume and little or no visible traction on the gel as detected by phase-contrast microscopy. The remaining cell types usually formed sheets on the surface of the gel; this was associated with an initial reduction in vitreous volume of approximately 50%, but little change thereafter. The cell mediated traction events on the structure of the vitreous gel were followed by time lapse video microscopy and by electron microscopy at various times after seeding of cells on the gels. The appearances corresponded closely with reported clinico-pathological studies of cellular proliferation in the vitreous. We believe that this in vitro model has several advantages over in vivo models of cellular proliferation in the vitreous, in that it permits analysis of individual cell behaviour and it is eminently suitable for pharmacologic manipulation.

Animals↗