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Characterization of phenolic compounds in virgin olive oil and their effect on the formation of carcinogenic/mutagenic heterocyclic amines in a model system.

Mutagenic heterocyclic amines (HAs) are formed at low levels during cooking of meat and fish, and some of them are considered to be possible human carcinogens. The formation of HAs may be affected by the presence of synthetic or naturally occurring antioxidants. In the present study the effect of virgin olive oil (VOO) phenolic compounds, identified and quantified by LC-MS, on the formation of HAs in a model system was evaluated. An aqueous solution of creatinine, glucose, and glycine was heated in the presence of two samples of VOO differing only in the composition of phenolic compounds. The addition of VOO to the model system inhibited the formation of 2-amino-3-methylimidazo[4,5-f]quinoxaline (IQx), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), and 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx) by between 30 and 50% compared with the control. Fresh-made olive oil, which contained a high amount of dihydroxyphenylethanol derivatives, inhibited HA formation more than a 1-year-old oil did. The inhibition of HA formation was also verified using phenolic compounds extracted from VOO.

Amines↗

Ecotoxicological assessment of bromobenzene using a test battery with five model systems.

Bromobenzene (BrB) is used as a solvent for crystallization and as an additive to motor oils and may be released into the environment through various waste streams. However, there is limited available information about the toxic hazard of BrB in the aquatic environment. Consequently, the ecotoxicological effects induced by BrB were investigated using five model systems with representants from four trophic levels. The battery included bioluminescence inhibition of the bacterium Vibrio fischeri, growth inhibition of the alga Chlorella vulgaris and immobilization of the cladoceran Daphnia magna. Total protein content, neutral red uptake and MTS metabolization were reduced, while lysosomal function, succinate dehydrogenase activity, G6PDH activity and leakage, metallothionein levels and EROD activity were stimulated in PLHC-1 and RTG-2 fish cell lines. The most sensitive bioindicator was the bioluminiscence of V. fischeri, with an EC(50) of 0.04mM BrB at 15min and a non-observed adverse effect level of 0.02 mM BrB. There is a large difference in sensitivity to BrB among the model systems probably due to the metabolic capacity of the different species. PLHC-1 cells were more sensitive to BrB than RTG-2 cells. The most prominent morphological effects observed were hydropic degeneration, loss of cells and of the perinuclear pattern of distribution of lysosomes. Therefore, BrB should be classified as toxic to aquatic organisms.

Aliivibrio fischeri↗

Single crystal EPR study of electronic structure and exchange interactions for copper(II)(L-arginine)2(SO4).(H2O)6: a model system to study exchange interactions between unpaired spins in proteins.

We report EPR measurements at 9.77 and 34.1 GHz in powder and single crystal samples of the ternary copper amino acid complex Cu(L-arginine)(2)(SO(4)).(H(2)O)(6). The single crystal Electron Paramagnetic Resonance spectra display a single resonance for all magnetic field orientations in the ca and cb crystal planes. In the ab plane they display two resonances for most orientations of the magnetic field, and only one resonance for orientations close to the crystal axes. This behavior is a result of the selective collapse of the resonances corresponding to the four copper sites in the unit cell produced by the exchange interactions between copper ions. From the characteristics of the collapse and the angular dependences of the position and width of the resonances we evaluate the g-tensors of the copper molecules and estimate exchange interactions |J(1)/k(B)|=0.9 K and |J(2)/k(B)|=0.009 K between copper neighbors at 5.908 A and at 15.684 A, respectively. J(1) is assigned to a syn-anti equatorial-apical carboxylate bridge with a total bond length of 7.133 A. J(2) is assigned to a long bridge of 12 atoms with a total bond length of 19.789 A, that includes two hydrogen bonds. The results are discussed in terms of the crystal and electronic structure of Cu(L-arginine)(2)(SO(4)).(H(2)O)(6). We show that J(2) is in excellent agreement with the observed magnetic interaction between the reduced quinone acceptors in the photosynthetic reaction center protein of the bacterium Rb. sphaeroides, which is transmitted along a similar chemical path containing two hydrogen bonds. Our findings indicate that it is valid to estimate values for the exchange interactions between redox centers in proteins transmitted along long chemical paths containing sigma and H-bonds, from data obtained in model systems, and emphasize the importance of measuring exchange interactions in biologically relevant model systems.

Arginine↗

Rat C6 glioma as experimental model system for the study of glioblastoma growth and invasion.

Infiltration of the central nervous system by neoplastic cells in patients with glioblastoma multiforme (GBM) leads to neurological dysfunction and eventually to death. The elucidation of the mechanisms underlying the aggressive nature of GBM aims at improving radio-, chemo- and gene therapy. This review is focused on the use of rat C6 glioma as an experimental model system for GBM and provides an overview of the experimental data published in the literature using this cell line in elucidating the mechanism of tumor growth, angiogenesis and invasion, and in the design and evaluation of anticancer therapies. Understanding the stages of malignant brain tumor progression requires a series of experimental approaches with a varying degree of complexity. Implantation of malignant cells into animal brain tissue closely resembles in vivo tumor growth and has the advantage over simplified models that inflammatory and vascular mechanisms are activated. However, the complexity of these models makes it difficult to identify the individual processes involved in sustained tumor growth, angiogenesis and invasion. In cell culture models, the effect of growth factors, extracellular matrix components, proteases and adhesion molecules can be investigated. The secretion of tumor-derived factors into the medium can also be analyzed when simplified models are used. This review is a compilation of experimental data focused on the characterization of tumor-related processes and on the evaluation of new therapies for the treatment of malignant glial neoplasms using rat C6 glioma as a model system.

Animals↗

Peptide model systems for amyloid fiber formation: design strategies and validation methods.

The rational understanding of the factors involved in the formation of amyloid deposits in tissue is fundamental to the identification of novel therapeutic strategies to prevent or cure pathological conditions such as Alzheimer's and Parkinson's disease or spongiform encephalopathies. Given the complexity of the molecular events driving protein self-association, a frequent strategy in the field has consisted of designing simplified model systems that facilitate the analysis of the elements that predispose polypeptides toward amyloid formation. In fact, these systems have provided very valuable knowledge on the determinants underlying structural transitions to the polymeric beta-sheet state present in amyloid fibers and more disordered aggregates. In this chapter, we will describe different approaches to obtain and design model systems for amyloidogenesis, as well as the methodologies that are typically used to validate them. We will also show how some of the general principles obtained from these studies can be applied for de novo design purposes and for the sequence-based identification of amyloidogenic stretches in proteins.

Alzheimer Disease↗

Model system to evaluate the effect of ampD mutations on AmpC-mediated beta-lactam resistance.

Mutations within the structural gene of ampD can lead to AmpC overproduction and increases in beta-lactam MICs in organisms with an inducible ampC. However, identification of mutations alone cannot predict the impact that those mutations have on AmpD function. Therefore, a model system was designed to determine the effect of ampD mutations on ceftazidime MICs using an AmpD(-) mutant Escherichia coli strain which produced an inducible plasmid-encoded AmpC. ampD genes were amplified by PCR from strains of E. coli, Citrobacter freundii, and Pseudomonas aeruginosa. Also, carboxy-terminal truncations of C. freundii ampD genes were constructed representing deletions of 10, 21, or 25 codons. Amplified ampD products were cloned into pACYC184 containing inducible bla(ACT-1)-ampR. Plasmids were transformed into E. coli strains JRG582 (AmpD(-)) and K-12 259 (AmpD(+)). The strains were evaluated for a derepressed phenotype using ceftazidime MICs. Some mutated ampD genes, including the ampD gene of a derepressed C. freundii isolate, resulted in substantial decreases in ceftazidime MICs (from >256 microg/ml to 12 to 24 microg/ml) for the AmpD(-) strain, indicating no role for these mutations in derepressed phenotypes. However, ampD truncation products and ampD from a partially derepressed P. aeruginosa strain resulted in ceftazidime MICs of >256 microg/ml, indicating a role for these gene modifications in derepressed phenotypes. The use of this model system indicated that alternative mechanisms were involved in the derepressed phenotype observed in strains of C. freundii and P. aeruginosa. The alternative mechanism involved in the derepressed phenotype of the C. freundii isolate was downregulation of ampD transcription.

Anti-Bacterial Agents↗

Application of watershed modeling system (WMS) for integrated management of a watershed in Turkey.

Watershed models, that enable the quantification of current and future pollution loading impacts, are essential tools to address the functions and conflicts faced in watershed planning and management. In this study, the Watershed Modeling System (WMS) version 7.1 was used for the delineation of boundaries of Koycegiz Lake-Dalyan Lagoon watershed located in the southwest of Turkey at the Mediterranean Sea coast. A Digital Elevation Model (DEM) was created for one of the major streams of the watershed, namely, Kargicak Creek by using WMS, and DEM data were further used to extract stream networks and delineate the watershed boundaries. Typical properties like drainage areas, characteristic length and slope of sub-drainage areas have also been determined to be used as model inputs in hydrological and diffuse pollution modeling. Besides, run-off hydrographs for the sub-drainages have been calculated using the Rational Method, which produces valuable data for calculating the time variable inflow and input pollution loads to be further utilized in the future water quality models of the Creek. Application of WMS in the study has shown that, it is capable to visualize the results in establishing watershed management strategies.

Environmental Monitoring↗

Single-system models and interference in category learning: commentary on Waldron and Ashby (2001).

In a recent article, Waldron and Ashby (2001) observed that performing a concurrent task caused greater interference in learning a simple one-dimensional categorization rule than in learning a complex three-dimensional one. They argued that this result was incompatible with all existing single-system models of category learning but was as predicted by the multiple-system COVIS model (Ashby, Alfonso-Reese, Turken, & Waldron, 1998). In contrast to Waldron and Ashby's argument, we demonstrate that the single-system ALCOVE model (Kruschke, 1992) naturally predicts the result by assuming that its selective-attention learning process is disrupted by the concurrent task.

Attention↗

Beta amyloid fragments derived from activated platelets deposit in cerebrovascular endothelium: usage of a novel blood brain barrier endothelial cell model system.

Amyloid precursor protein (A betaPP) processing results in generation of amyloid beta peptide (A beta) which deposits in the brain parenchyma and cerebrovasculature of patients with Alzheimer's disease (AD). Evidence that the vascular deposits derive in part from A betaPP fragments originating from activated platelets includes findings that individuals who have had multiple small strokes have a higher prevalence of AD compared to individuals who have taken anti-platelet drugs. Thus, determination of whether platelet A betaPP fragments are capable of traversing the blood-brain barrier (BBB) is critical. We have established that activated platelets from patients with AD retain more surface transmembrane-bound A betaPP (mA betaPP) than control platelets. We report here that this mA betaPP can be cleaved to A beta-containing fragments which pass through a novel BBB model system. This model utilizes human BBB endothelial cells (BEC) isolated from brains of patients with AD. These BEC, after exposure to activated platelets which have been surface-labeled with fluorescein and express surface-retained mA betaPP, cleave fluorescein-tagged surface proteins, including mA betaPP, resulting in passage to the BEC layer The data confirm that BEC contribute to processing of platelet-derived mA betaPP and show that the processing yields A beta containing fragments which could potentially contribute to cerebrovascular A beta deposition.

Adult↗

Transfected Chinese hamster ovary cells as model system for cytokine immunocytochemistry and in situ hybridisation.

The presence of cytokine producing cells is most easily revealed by techniques measuring the secreted cytokines in culture supernatants or body fluids. However, these techniques only measure the bulk cytokine release by a given, often mixed cell population. To demonstrate cytokine production at the single cell level, immunocytochemistry (ICC) and in situ hybridisation (ISH) are now widely used techniques. To establish these techniques, an easily accessible model system is needed which permits the evaluation of different ICC and ISH protocols. It can be used to demonstrate the specificity of the antibodies and may serve as a positive control for samples of unknown cytokine content. Here we propose the use of Chinese hamster ovary (CHO) cells transfected to express one specific cytokine as such a model system. Its usefulness is demonstrated by the characterisation of six monoclonal antibodies to human interleukin-4 and the establishment of two in situ hybridisation protocols.

Animals↗

Transfected Chinese hamster ovary cells as a model system for cytokine immunocytochemistry and in situ hybridisation.

The presence of cytokine producing cells is most easily revealed by techniques measuring the secreted cytokines in culture supernatants or body fluids. However, these techniques only measure the bulk cytokine release by a given, often mixed cell population. To demonstrate cytokine production at the single cell level, immunocytochemistry (ICC) and in situ hybridisation (ISH) are now widely used techniques. To establish these techniques, an easily accessible model system is needed which permits the evaluation of different ICC and ISH protocols. It can be used to demonstrate the specificity of the antibodies and may serve as a positive control for samples of unknown cytokine content. Here we propose the use of Chinese hamster ovary (CHO) cells transfected to express one specific cytokine as such a model system. Its usefulness is demonstrated by the characterisation of six monoclonal antibodies to human interleukin-4 and the establishment of two in situ hybridisation protocols.

Animals↗

Mixtures of a series of homologous hydrophobic peptides with lipid bilayers: a simple model system for examining the protein-lipid interface.

The interactions of several members of a homologous series of peptides with the phospholipid bilayer have been examined by using fluorescence and deuterium NMR spectroscopy, differential scanning calorimetry, and measurements of water-to-bilayer partition coefficients. 1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers and tripeptides of the form Ala-X-Ala-O-tert-butyl are used as a model system to probe the influence of amino acid side-chain substitution on the insertion of peptides into membranes and the behavior of peptide/bilayer mixtures. Tripeptides with X = Gly, Ala, Phe, and Trp have been examined. All of the tripeptides are water soluble, and all partition into DMPC bilayer vesicles to some extent. The Gly-containing peptide is the least soluble and the Trp-containing peptide the most soluble in the bilayer. The extent of perturbation of the bilayer structure induced by the peptides parallels their bilayer solubility: the Gly and Ala peptides act as simple impurities while peptides containing bulky aromatic rings cause a phase separation. Changes in the fluorescence properties of the Trp analogue upon incorporation into the bilayer indicate that the Trp side chain is probably immersed in the hydrocarbon region of the bilayer. Peptides of this form should serve as easily modifiable model systems with which to examine details of how the bilayer environment affects peptide conformation, as well as how hydrophobic peptides affect the bilayer structure.

Calorimetry, Differential Scanning↗

Simulation program for optimal orthopedic call: a modeling system for orthopedic surgical trauma call.

This report uses a mathematical modeling system to define optimal orthopedic coverage for trauma centers. Data from 2,325 patients treated with emergency orthopedic operations within 24 hours of admission at 78 randomly sampled and at four totally sampled verified centers were used to create a profile of (1) admission by month, day, and hour; (2) operation times; and (3) operation duration. The reason for operation included (1) open fracture or crush (809 patients); (2) irreducible dislocations (164 patients); (3) fracture with vascular injury (seven patients); (4) dislocation with vascular injury (17 patients); (5) compartment syndrome (11 patients); (6) femoral neck fracture in young patients (36 patients); (7) combination of categories 1 to 6 (70 patients); (8) fracture with multiple injuries (171 patients); and (9) urgent not emergent (1,040 patients). The program defined the frequency that an injured patient needing an orthopedic consult would wait beyond 30 minutes because the orthopedic surgeon was doing a trauma related operation at a center with one or two orthopedic surgeons on call. The probability that a patient cannot be seen promptly by one orthopedic surgeon in a center doing 25, 50, 75, 100, 200, and 300 emergency procedures per year is 0.17, 0.74, 1.6, 3.1, 12.5, and 28 patients per year. When two are on call, 1.3 patients, yearly, will wait more than 30 minutes in a center doing 300 emergency procedures. Thus, mandatory orthopedic backup call for a trauma center performing fewer than 100 emergent trauma procedures within 24 hours is unwarranted.

Computer Simulation↗

Interactions of human ovarian tumor cells with human mesothelial cells grown on extracellular matrix. An in vitro model system for studying tumor cell adhesion and invasion.

Human ovarian tumors metastasize by direct extension into the peritoneal cavity leading to tumor cell implantation onto peritoneal surfaces. Successful formation of peritoneal implants is dependent on the ability of ascitic tumor cells to infiltrate the mesothelium, and become firmly adherent to the underlying extracellular matrix (ECM). In order to investigate this process in more detail, an in vitro model system was developed employing human mesothelial cells grown on ECM-coated culture dishes. The ability of human ovarian carcinoma cells derived from ascitic fluid to attach to the mesothelial cell monolayer grown on ECM, ECM alone or plastic was quantitated with the use of 51Cr radio-labelled tumor cells. Tumor cells exhibited a more rapid and firmer attachment to ECM than to the mesothelial cells or to plastic. Using agitation to stimulate peritoneal fluid dynamics and shear forces in vivo, tumor cell arrest was found to be limited to the ECM, but it occurred at a slower rate than it did without agitation. Tumor cell attachment was also restricted to areas of exposed ECM in wounded mesothelium as assessed by phase-contrast microscopy. Morphologic alterations of the mesothelium induced by tumor cells were observed with the use of scanning electron microscopy (SEM) and immunohistochemical staining which included disruption of intercellular junctions leading to retraction of mesothelial cells, exposure of underlying ECM, subsequent attachment and proliferation on ECM. This model system would appear to be useful for elucidating mechanisms of ovarian tumor cell adhesion and proliferation, and for assessing various therapeutic modalities for their ability to block tumor cell implantation, invasion and growth on peritoneal surfaces.

Animals↗

A model system for tumor angiogenesis: involvement of transforming growth factor-alpha in tube formation of human microvascular endothelial cells induced by esophageal cancer cells.

Tumor growth is dependent on angiogenesis, which is thought to be mediated through growth factors, such as transforming growth factor-alpha (TGF-alpha) and -beta (TGF-beta), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF), produced by tumor cells. We have developed a model system for tumor angiogenesis in vitro: tube formation of human omentum microvascular endothelial (HOME) cells in type I collagen gels when these cells are co-cultured with tumor cells. Exogenously added TGF-alpha induced tube formation of HOME cells in collagen gel. In contrast, TGF-beta inhibited the TGF-alpha-induced tube formation of endothelial cells. We investigated whether tube formation could be induced in HOME cells in collagen gel when the HOME cells were co-cultured with three esophageal cancer cell lines, TE1, TE2, and TE5. TE1 and TE2 cells expressed both TGF-alpha and TGF-beta mRNA, but the level of TGF-alpha mRNA in TE2 was found to be much lower than in TE1 cells. TE5 did not express either TGF-alpha or TGF-beta. The tube formation of HOME cell was induced when they were co-cultured with TE1 cells, while both TE2 and TE5 cell lines induced tube formation at much lower rates than TE1. TE1-induced tube formation of HOME cells was specifically blocked by co-administration of anti-TGF-alpha-antibody, but not by anti-bFGF-antibody. The present study suggests that, in our model system, esophageal tumor angiogenesis is partly controlled by TGF-alpha, possibly through a paracrine pathway.

Blotting, Northern↗

A model system to study the environment-dependent expression of the Bet v 1a gene encoding the major birch pollen allergen.

BACKGROUND: The major birch pollen allergen Bet v 1 (or Bet v 1a) is one of the main causes of seasonal type I allergies. Various environmental factors such as light, temperature and air pollution may influence the activity of the Bet v 1a gene. The creation of a model system to evaluate the role of environmental factors affecting the Bet v 1a gene expression would be highly desirable. We suggest the use of transgenic tobacco plants carrying a Bet v 1a promoter-reporter gene fusion as such a system. METHODS: The promoter of the Bet v 1a gene was isolated with the use of the Universal Genome Walker kit (BD Biosciences Clontech, USA). Web Software was used to search for putative cis-regulatory elements within the promoter. Transgenic tobacco plants harboring the promoter-beta-glucuronidase (GUS) reporter gene fusion were obtained via Agrobacterium tumefaciens-mediated transformation. Promoter activity was examined with histochemical and quantitative assays. RESULTS: Structural analysis predicted elements responsible for pollen-specific, light-, stress- and hormone-mediated induction within the Bet v 1a promoter. The evaluation of GUS activity in transgenic tobacco plants showed that the Bet v 1a promoter is pollen-specific. Moreover, the Bet v 1a promoter is considered to be the strongest isolated pollen-specific promoter reported to date. It was shown that temperature and abscisic acid positively regulate the activity of the Bet v 1a promoter during pollen development, providing evidence for environment-dependent regulation of the Bet v 1a gene. CONCLUSIONS: A model system to study the effect of environmental factors on the expression of the Bet v 1a gene encoding the major birch allergen in pollen was generated. Additionally, we suggest that this system could be used to search for factors that inhibit the activity of the gene in pollen in order to reduce the potential allergenicity of birch trees.

Abscisic Acid↗

Role analysis of the advanced practice nurse using the Neuman Health Care Systems Model as a framework.

The role of the advanced practitioner of nursing is perceived and operationalized differently from practice setting to practice setting. These differences should compel practitioners to analyze and differentiate role expectations within specific practice situations. The Neuman Health Care Systems Model provides a systematic framework for role analysis, development, enactment, and evaluation. The analysis should serve to decrease role stress, strain, and ambiguity while ultimately increasing role implementation success. The purposes of this article are: (a) to describe the general systems theory that is the basis of the Neuman model, (b) to describe the Neuman Health Care Systems Model to provide a basis for role analysis, and (c) to provide a structure for role analysis of the CNS. The benefit of a systematic microanalysis of the role of the CNS will be to develop a template to be used in the analysis of a particular position in the real world.

Humans↗

Three-dimensional computer modelling system for the study of biological structures.

A three-dimensional computer modelling system has been developed for use in biology, and is currently running on a Sun3 computer. The data originate as a series of two-dimensional micrographs which are digitised via a TV camera. The two-dimensional images are used to select features of interest and to construct a three-dimensional model. This model can be viewed in vector or solid format, it can be rotated about three orthogonal axes and can be viewed in three dimensions as a stereo pair or an anaglyph. The system has been used in a large number of projects over the past 10-15 years, for example, to examine physiological and nerve structures. The time-consuming part of the process is the selection of features, which involves a high level of biological expertise. Present developments are concerned with reduction of the time spent in feature recognition and involve the introduction of expert systems together with human-computer interaction to deal with problems of identification.

Computer Simulation↗