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Resonance energy transfer in a model system of membranes: application to gel and liquid crystalline phases.

Resonance energy transfer between octadecyl rhodamine B (donor) and 1,1',3,3,3',3'-hexamethylindotricarbocyanine (acceptor) was studied in a model system of membranes (large unilamellar vesicles of dipalmitoylphosphatidylcholine), using both steady-state and time-resolved techniques. In the fluid phase (temperature = 50 degrees C) the decay law and the steady-state theoretical curve for energy transfer in two dimensions are verified. In the gel phase (temperature = 25 degrees C) an apparent reduction of dimensionality is observed, which is explained on the basis of probe segregation to the defect lines (grain boundaries). An estimation of the domain size from the model recovered linear probe concentrations is approximately 1750-2000 lipid molecules. In both phases, the existence of a fractal geometry was ruled out.

1,2-Dipalmitoylphosphatidylcholine↗

The development of animal model systems for HIV-1 encephalitis and its associated dementia.

The human immunodeficiency virus (HIV) is neuroinvasive and can be neurovirulent. Indeed, 20-30% of individuals with the acquired immune deficiency syndrome (AIDS) develop cognitive and motor dysfunction (termed the AIDS dementia complex or HIV dementia) coincident with advanced immunosuppression. Despite massive research efforts to discern viral neuropathogenic mechanisms, much remains incompletely understood. Recently, we and others developed animal model systems to elucidate how HIV infection within the brain can lead to impairment of central nervous system function. In this report, we evaluate each of the published animal models for their ability to mirror HIV dementia. Ease of handling and expense were also under consideration. Ultimately, studies in animal systems should permit a better understanding of the nature of HIV-1-induced neurological injury and aid in the development of effective treatments for this dreaded complication of HIV infection.

AIDS Dementia Complex↗

H-ras activation in benign and self-regressing skin tumors (keratoacanthomas) in both humans and an animal model system.

The involvement of the ras oncogenes in tumorigenesis was investigated in keratoacanthomas, which are benign and self-regressing skin tumors, both in humans and in a corresponding animal model system. Keratoacanthomas were induced on rabbit ears by repeated applications of 7,12-dimethylbenz(a)anthracene. About 60% of the tumor DNAs produced transformed foci after transfection into NIH 3T3 cells, and in all of them the transforming gene was identified as H-ras by Southern and Northern (RNA) hybridization. Immunoprecipitation experiments suggested that the transforming rabbit H-ras protein carried a mutation in codon 61. In addition, an activated H-ras gene was detected in a human keratoacanthoma by using a nude mouse tumorigenesis assay after transfection of tumor DNA into NIH 3T3 cells. This is the first report of ras activation in a benign human tumor. The transforming human H-ras gene showed a point mutation in codon 61 that would result in leucine instead of the glutamine present in the normal gene product. The finding of ras activation in tumors that are not only benign but also self-regressing indicates that activated ras genes are not sufficient to maintain a neoplastic phenotype, although they likely play a role in early stages of tumorigenesis.

9,10-Dimethyl-1,2-benzanthracene↗

Carcinogen metabolism in human skin grafted onto athymic nude mice: a model system for the study of human skin carcinogenesis.

Human skin grafted onto athymic nude mice maintains its major histological features and may provide a useful system with which to assess the carcinogen interaction with human skin. Significant differences were observed in basal levels of cytochrome P-450 and cytochrome P-448-dependent monooxygenase activities between human grafted and nude mouse epidermis. Topical application of crude coal tar (CCT) to human skin transplanted onto nude mice resulted in 3.9 & 3.5; 3.2 & 2.9 and 1.1 & 1.2 fold increases in mouse and human epidermal aryl hydrocarbon hydroxylase (AHH), ethoxyresorufin deethylase (ERD) and ethoxycoumarin deethylase (ECD) activities, respectively. CCT applied topically to mouse skin resulted in 27.8 & 6.4; 12.8 & 3.3 and 1.7 & 2.6 fold increases in mouse and human epidermal AHH, ERD and ECD activities, respectively. Topical application of coal tar either onto human transplanted skin or to mouse skin also resulted in substantial induction of hepatic and pulmonary AHH and ERD activities. These studies indicate that human skin grafted onto nude mice preserves its metabolic capacity and offers a useful model system with which to assess the effects of polycyclic aromatic hydrocarbons and CCT on cutaneous xenobiotic metabolism in the human population.

7-Alkoxycoumarin O-Dealkylase↗

Autoradiography and immunofluorescence combined for autecological study of single cell activity with Nitrobacter as a model system.

Specific detection of a particular bacterium by immunofluorescence was combined with estimation of its metabolic activity by autoradiography. The nitrifying bacteria Nitrobacter agilis and N. winogradskyi were used as a model system. Nitrobacter were incubated with NaH14CO3 and 14CO2 prior to study. The same preparations made for autoradiograms were stained with fluorescent antibodies specific for the Nitrobacter species. Examination by epifluorescence and transmitted dark-field microscopy revealed Nitrobacter cells with and without associated silver grains. Direct detection and simultaneous evaluation of metabolic activity of Nitrobacter was demonstrated in pure cultures, in a simple mixed culture, and in a natural soil.

Autoradiography↗

It takes guts: the Drosophila hindgut as a model system for organogenesis.

The Drosophila hindgut is fruitful territory for investigation of events common to many types of organogenesis. The development of the Drosophila hindgut provides, in microcosm, a genetic model system for studying processes such as establishment (patterning) of an epithelial primordium, its internalization by gastrulation, development of left--right asymmetric looping, patterning in both the anteroposterior and dorsoventral axes, innervation, investment of an epithelium with mesoderm, reciprocal epitheliomesenchymal interactions, cell shape change, and cell rearrangement. We review the genetic control of these processes during development of the Drosophila hindgut, and compare these to related processes in other bilaterians, particularly vertebrates. We propose that caudal/Cdx, brachyenteron/Brachyury, fork head/HNF-3, and wingless/Wnt constitute a conserved "cassette" of genes expressed in the blastopore and later in the gut, involved in posterior patterning, cell rearrangement, and gut maintenance. Elongation of the internalized Drosophila hindgut primordium is similar to elongation of the archenteron and also of the entire embryonic axis (both during and after gastrulation), as well as of various tubules (e.g., nephric ducts, Malpighian tubules), as it is driven by cell rearrangement. The genes drumstick, bowl, and lines (which encode putative transcriptional regulators) are required for this cell rearrangement, as well as for spatially localized gene expression required to establish the three morphologically distinct subregions of the hindgut. Expression of signaling molecules regulated by drumstick, bowl, and lines, in particular of the JAK/STAT activator Unpaired at the hindgut anterior, may play a role in controlling hindgut cell rearrangement. Other cell signaling molecules expressed in the hindgut epithelium are required to establish its normal size (Dpp and Hh), and to establish and maintain the hindgut visceral mesoderm (Wg and Hh). Both maternal gene activity and zygotic gene activity are required for asymmetric left--right looping of the hindgut. Some of the same genes (caudal and brachyenteron) required for embryonic hindgut development also act during pupation to construct a new hindgut from imaginal cells. Application of the plethora of genetic techniques available in Drosophila, including forward genetic screens, should identify additional genes controlling hindgut development and thus shed light on a variety of common morphogenetic processes.

Animals↗

Maximum-likelihood versus maximum a posteriori parameter estimation of physiological system models: the C-peptide impulse response case study.

Maximum-likelihood (ML), also given its connection to least-squares (LS), is widely adopted in parameter estimation of physiological system models, i.e., assigning numerical values to the unknown model parameters from the experimental data. A more sophisticated but less used approach is maximum a posteriori (MAP) estimation. Conceptually, while ML adopts a Fisherian approach, i.e., only experimental measurements are supplied to the estimator, MAP estimation is a Bayesian approach, i.e., a priori available statistical information on the unknown parameters is also exploited for their estimation. In this paper, after a brief review of the theory behind ML and MAP estimators, we compare their performance in the solution of a case study concerning the determination of the parameters of a sum of exponential model which describes the impulse response of C-peptide (CP), a key substance for reconstructing insulin secretion. The results show that MAP estimation always leads to parameter estimates with a precision (sometimes significantly) higher than that obtained through ML, at the cost of only a slightly worse fit. Thus, a three exponential model can be adopted to describe the CP impulse response model in place of the two exponential model usually identified in the literature by the ML/LS approach. Simulated case studies are also reported to evidence the importance of taking into account a priori information in a data poor situation, e.g., when a few or too noisy measurements are available. In conclusion, our results show that, when a priori information on the unknown model parameters is available, Bayes estimation can be of relevant interest, since it can significantly improve the precision of parameter estimates with respect to Fisher estimation. This may also allow the adoption of more complex models than those determinable by a Fisherian approach.

Bayes Theorem↗

Effects of seasonings on the stability of ascorbic acid in a cooking model system.

The thermolability of ascorbic acid (AA) in aqueous solution at 100 degrees C was assessed in the presence of various seasonings commonly used in Japanese-style cooking. A model system approximated Japanese cooking with regard to the concentrations of AA and seasonings and the heating time. The decrease of AA in the reaction system of this experiment was a first-order reaction with respect to the concentration of AA loss. Although kinetic constants for AA loss decreased with increasing concentrations of AA (25-400 microg/mL), the absolute amounts degraded were almost the same for all AA concentrations, suggesting that dissolved oxygen is one of main factors affecting the stability of AA solutions during heating at 100 degrees C. When each seasoning was added to AA solution, salt stabilized AA and Japanese alcohol-containing admixtures, such as sake and sweet sake (mirin), did not have a significant effect on the stability. Conversely, soy sauce, miso (fermented soybean paste) and broth powder from skipjack accelerated the decrease of AA in a concentration-dependent manner. The kinetic study suggested that oxygen was rapidly consumed and AA loss accelerated by addition of soy sauce or miso to AA solution. Consequently it is likely that a reaction mechanism shifts from aerobic to anaerobic and the forward reactions proceed. Of the constituents of Japanese seasonings, not only iron but also amino acids are involved in the acceleration of AA degradation. The presence of amino acids should be taken into account when considering the levels of AA in soups.

Amino Acids↗

Developing a plan for measuring outcomes in model systems of care for American Indian and Alaska Native children and youth.

The Circles of Care initiative emphasized the importance of developing an outcomes measurement plan that was consonant with the model system of care as well as community values and priorities. This analysis suggests that the Circles of Care grantees achieved this key programmatic objective, but that a major constraint was the tendency of funders, including the Substance Abuse and Mental Health Services Administration (the funder of Circles of Care), to mandate their own outcomes measurement plans. Funders are encouraged to balance their needs for commonality of measures across programs for their own evaluation purposes with the needs of service providers to utilize measures that meet their unique programmatic and community contexts.

Adolescent↗

Raman study of potential "antisense" drugs: nonamer oligonucleotide duplexes with a central mismatch as a model system for the binding selectivity evaluation.

A set of four 9-mer oligonucleotide duplexes formed between the 5'-GCATNTCAC-3', N=A,C,T,G, and the 5'-GTGATATGC-3' complement has been proposed as a model system for the investigation of novel oligonucleotide analogues (candidates for antisense use) binding selectivity. Raman measurements were carried out on a set of natural DNA 9-mer in order to verify suitability of the model and to obtain reference spectral data. Difference Raman spectra between the mismatch and match duplexes obtained at 15 degrees C exhibited numerous spectral features sensitively indicating the structural changes. All the three mismatches only very weakly disturb the overall B-form conformation of the duplex. Significant structural changes that occurred at the mismatch site are reflected mainly by the neighboring thymidine Raman bands at 1377, 1650, and 1675 cm(-1). The intensity change of the two latter bands is almost the same for the T:G and the T:T mismatch while in the case of the T:C mismatch it is just opposite, demonstrating a very different arrangement of the mismatched pair.

Base Pair Mismatch↗

Model-based administration of inhalation anaesthesia. 3. Validating the system model.

BACKGROUND: We quantified the predictive performance of our computer model of the administration of inhalation anaesthesia from a Datex-Ohmeda Modulus CD circle-absorber system. METHODS: In 50 patients, desflurane anaesthesia was maintained with a fresh-gas flow (FGF) of 0.5 litres min(-1) of both nitrous oxide and oxygen, preceded by fast (n=14) or slow (n=36) induction: FGF greater than total ventilation, Group F; FGF equal to 1.0 litres min(-1), Group S. The two versions of the model studied differed in the size of their inter-tissue diffusion, as 0 (version 1) and 3% (version 2) of the cardiac output was shifted from the viscera to adipose tissue. Model performance was judged by comparing measured and predicted gas concentrations in terms of three variables for each gas concentration in each patient: root mean squared error (rmse=total error), bias (mean predicted - measured) (systematic error), and scatter (error around bias). These variables were then averaged over all patients. These measures were calculated overall, and separately for each group and each stage (1 = induction or 2 = maintenance). RESULTS: Model predictions were in reasonable to very good agreement with clinically obtained data. Version 2 performed better than version 1. Differences between groups were not demonstrated. The model performed better for stage 2, but only for desflurane. In group S, results (mean (SD); as percentages of the measured values for nitrous oxide, oxygen and desflurane) in the order rmse, bias, and scatter were for end-tidal concentrations of nitrous oxide: 8 (4), 8 (5), 2 (1)%; oxygen: 11 (4), -10 (6), 2 (1.1)%; nitrogen: 0.9 (0.6), -0.8 (0.6), 0.2 (0.1) vol%; carbon dioxide: 1.8 (0.6), 1.8 (0.6), 0.2 (0.1) vol%; desflurane, stage 2: 8 (4), 4 (7), 4 (2)%, vs 15 (6), -10 (8), 9 (4)% for stage 1. CONCLUSION: Administration of inhalation anaesthesia can be based on version 2 of this model, but must be guided by active monitoring.

Adolescent↗

Investigations on the promoting effect of ammonium hydrogencarbonate on the formation of acrylamide in model systems.

NH4HCO3 is known to promote acrylamide formation in sweet bakery products. This effect was investigated with respect to sugar fragmentation and formation of acrylamide from asparagine and sugar fragments in model systems under mild conditions. The presence of NH4HCO3 led to increases in acrylamide and alpha-dicarbonyls from glucose and fructose, respectively. As compared to glucose or fructose, sugar fragments such as glyoxal, hydroxyethanal, and glyceraldehyde formed much higher amounts of acrylamide in reaction with asparagine. The enhancing effect of NH4HCO3 is explained by (1) the action of NH3 as base in the retro-aldol reactions leading to sugar fragments, (2) facilitated retro-aldol-type reactions of imines in their protonated forms leading to sugar fragments, and (3) oxidation of the enaminols whereby glyoxal and other reactive sugar fragments are formed. These alpha-dicarbonyl and alpha-hydroxy carbonyl compounds may play a key role in acrylamide formation, especially under mild conditions.

Acrylamide↗

An epistatic mating system model can produce permanent cytonuclear disequilibria in a hybrid zone.

We examine the evolutionary dynamics of gametic and genotypic disequilibria between a cytoplasmic gene and a nuclear gene under two mating system models relevant to hybrid zones. In the first model, in which female mating preference is determined by an epistatic interaction between the two loci, permanent nonzero cytonuclear disequilibria are possible for a variety of initial genotype frequencies, particularly when rates of assortative mating for the two parental species are high. In contrast, when mating preference is effectively determined by interaction between a cytoplasmic gene and the multilocus nuclear genotypes characteristic of the parental species, all cytonuclear disequilibria, as well as frequencies of pure parentals, rapidly decay to zero unless assortative mating is nearly perfect. Results of the models are applied to the interpretation of observed cytonuclear associations in a hybrid population of Hyla tree frogs.

Algorithms↗

[Myocardium of mdx mice contains factor(s) that damage DNA structure and retard DNA reparation after gamma-irradiation (experiences in modeling system)].

We studied the action of saline extracts of ventricle myocard (EM) of C57BL and mdx mice on DNA structure and repair of one-strand breaks of DNA in a modelling system. The system involves DNA repair in E. coli WP2 cells after gamma-irradiation. Using standard technique, DNA reparation was estimated on measuring the speed of E. coli DNA sedimentation in alkaline sucrose gradients. It was shown, that EM of C57BL or mdx mice exerted no influence on DNA repair, which was completely declined within 60 min with EM present in the growth medium of permeabilized E. coli. Addition of C57BL mice EM into lytic solution does not accelerate DNA sedimentation of nonirradiated E. coli. At the same time, EM of mdx mice sharply accelerates DNA sedimentation of nonirradiated E. coli reducing DNA molecular weight from 200 x 10(6) to 135 x 10(6) Da. At entering in the lytic solution the EM of mdx mice also slows down E. coli DNA repair after gamma-irradiation. It is supposed, that EM of mdx mice may contain a factor(s) damaging DNA in the E. coli lysate and presumably slowing down DNA reparation after gamma-irradiation. Russian Foundation of Basic Research Grants 99-04-49390, 02-04-49870 and 00-04-49390.

Animals↗

A model system for studying covalent binding of food carcinogens MeIQx, MeIQ and IQ to DNA and protein.

We have studied the covalent binding of carcinogenic aminoimidazoazaarene compounds to macromolecules in a microsomal model system. The 14C-labelled compounds were incubated with rat-liver microsomes, and binding to macromolecules was measured after their precipitation on glass filters, which were washed several times in organic solvents. The amount of radioactivity was determined by liquid scintillation counting. Covalent binding was dependent on the addition of NADPH, with an optimal concentration of about 1 mM. The binding appeared to follow saturation kinetics when carcinogen concentrations were lower than 200 microM, with Km values of less than 20 microM. At 50 microM, 2-amino-3,4-dimethylimidazo[4,5-f]-quinoline (MeIQ) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) bound more effectively than 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx). When DNA was included in the incubations, binding to this macromolecule was ten-fold less per milligram than binding to proteins. In comparison with microsomes from untreated animals, those from rats treated with beta-naphthoflavone caused up to nine-fold more binding of MeIQx, six-fold more of IQ and three times as much of MeIQ. Induction by Aroclor 1254 caused up to 17-fold more binding, whereas induction by phenobarbital caused up to three-fold more binding. The effects of the inducers were greatest for MeIQx and IQ, while smaller effects were seen for MeIQ. The results are most consistent with cytochrome P450-dependent metabolic activation of the carcinogens to hydroxylamine metabolites, for which an isoenzyme(s) inducible by polyaromatic and polychlorinated hydrocarbons is most effective. To our knowledge, this is the first report that MeIQx is metabolized to reactive species capable of covalent binding to macromolecules.

Animals↗

[Psychiatric management of mentally handicapped patients. A systemic model].

The care for mentally handicapped subjects in a geographically, linguistically and socioculturally well defined region of Switzerland (the region "Oberwallis") is described. The care and the treatment are following a systemic model. The mentally handicapped live outside the psychiatric hospital in social integrated housing and working structures, which are established by an autonomous organized association. There exists a close cooperation between this association and the psychiatric hospital, which is integrated in a general hospital. Both institutions, the common theoretical basis and forms of cooperation are described.

Activities of Daily Living↗

Computer-assisted corneal topography: accuracy and reproducibility of the topographic modeling system.

BACKGROUND: Assessment of the efficacy and stability of novel refractive surgery techniques and comparative studies require accurate and reproducible corneal mapping technology, ideally to +/- 0.25 D. A vertically mounted Topographic Modeling System (TMS) system was evaluated. MATERIALS AND METHODS: Six calibrated PMMA spheres (30.00, 35.00, 43.00, 50.00, 55.00, and 60.00 D) were used. Two examiners performed three measurements on each test sphere and compared appropriate data with a previously tested and calibrated digital auto-keratometer (SK-1, precision < 0.25 D). The accuracy and reproducibility of all 25 rings of the TMS were studied as a function of focus (250 to 500 microns), centering (100 to 300 microns), and integrity of the inner and outer reflected placido images to mimic tear breaks at the corneal apex and peripheral vignetting and shadows produced by eyelids and eyelashes. RESULTS: The accuracy of the 25 rings decreased from the center to the periphery. The deviation scores (mean difference of three readings between measured and calibrated surface powers) of rings 3 through 25 were 100% within +/- 0.25 D only for the 43.00-diopter sphere. They were 56%, 68%, 43%, 13%, and 17% within +/- 0.25 D respectively for the 30.00-, 35.00-, 50.00-, 55.00-, and 60.00-diopter spheres. For 500 microns of defocusing, we observed a mean deviation of 0.35 D for the flattest sphere to 2.30 D for the 60.00-diopter sphere. For the same amount of defocusing, the error produced by the TMS increased progressively with the sphere dioptric power. Masking the first four and six rings, one by one, showed topographic maps to be valid only when all four central placido images were fully read by the computer. CONCLUSIONS: The TMS clinical performance could be improved by 1) increasing focus depth of field, 2) blocking data processing when central rings are incomplete or missing, 3) increasing the accuracy over the 30.00- to 60.00-diopter range, and 4) providing the user with calibration spheres.

Artifacts↗