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Monoclonal antibodies for the depletion of specific subpopulations of lymphocytes.

A number of rat monoclonal antibodies of the IgG2b subclass have been used to deplete mice of T lymphocyte subsets. It has been possible to produce long-term depletion where antibodies are administered to mice thymectomized in their adult life, or short-term depletion in euthymic animals. It is therefore feasible to ablate a T lymphocyte subpopulation at any stage in the course of an immune response and to examine in detail the role of a particular subset in the induction or effector phases of that response. We have used such ablative procedures to define the T cell subsets which participate in graft rejection, graft-versus-host disease, antigenic competition and antiviral and anti-self immunity and have attempted to exploit such knowledge to establish immunological tolerance in an adult animal.

Animals↗

Effects of the mutation glycine-222----aspartic acid on the functions of elongation factor Tu.

We have studied the properties of a mutant elongation factor Tu, encoded by tufB (EF-TuBo), in which Gly-222 is replaced by Asp. For its purification from the kirromycin-resistant EF-Tu encoded by tufA (EF-TuAr), a method was developed by exploiting the different affinities to kirromycin of the two factors and the competition between kirromycin and elongation factor Ts (EF-Ts) for binding to EF-Tu. The resulting EF-TuBo kirromycin and EF-TuAr EF-Ts complexes are separated by chromatography on diethylaminoethyl-Sephadex A-50. For the first time we have succeeded in obtaining a tufB product in homogeneous form. Compared with wild-type EF-Tu, EF-TuBo displays essentially the same affinity for GDP and GTP, with only the dissociation rate of EF-Tu GTP being slightly faster. Protection of amino-acyl-tRNA (aa-tRNA) against nonenzymatic deacylation by different EF-Tu species indicates that conformational alterations occur in the ternary complex EF-TuBo GTP aa-tRNA. However, the most dramatic modification is found in the EF-TuBo interaction with the ribosome. Its activity in poly(Phe) synthesis as well as in the GTPase activity associated with the interaction of its ternary complex with the ribosome mRNA complex requires higher Mg2+ concentrations than wild-type EF-Tu (Mg2+ optimum at 10-14 vs. 6 mM), even if EF-TuBo can sustain enzymatic binding of aa-tRNA to ribosomes at low Mg2+. The anomalous behavior of EF-TuBo is reflected in a remarkable increase of the fidelity in poly(Phe) synthesis, especially at high Mg2+ concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Aspartic Acid↗

Demand-based thiolate anion generation under virtually neutral conditions: influence of steric and electronic factors on chemo- and regioselective cleavage of aryl alkyl ethers.

Thiolate anions have been generated in a "demand-based" fashion under virtually neutral conditions for chemoselective deprotection of aryl alkyl ethers. Solvents play the critical role in making the reaction effective and should have high values of epsilon (>30), molecular polarizabilities (>10), and DN (>27) and low values of AN (<14). However, it is the combined effect of all of these physical properties that make a particular solvent effective. The reaction rates of cleavage of various aryl alkyl ethers are dependent on the steric crowding around the O-alkyl carbon and follow the order propargyl approximately allyl approximately benzyl > methyl > ethyl. Electron-withdrawing substituents increase the rate of ether cleavage reaction. The influence of the steric and electronic factors have been successfully exploited for selective deprotection of aryl alkyl ethers during inter- and intramolecular competitions.

Journal Article↗

Thiol metabolism of the trypanosomatids as potential drug targets.

Trypanosomatids produce significant amounts of four major low molecular mass thiols, trypanothione, glutathionylspermidine, glutathione, and ovothiol A. Of these, only glutathione is present in cells of the host. All four low molecular mass thiols are directly or indirectly maintained in a reduced state by trypanothione reductase. Available evidence, from gene disruption studies, indicate that this is an essential enzyme. Attempts to exploit trypanothione reductase as a chemotherapeutic target lead to the design of competitive and irreversible inhibitors of the enzyme. A promising route involves the design of redox cyclers interacting specifically with trypanothione reductase as subversive substrates. Progress in studies on the biosynthesis of ovothiol A is summarized.

Animals↗

Non-gridded library: a new approach for BAC (bacterial artificial chromosome) exploitation in hexaploid wheat (Triticum aestivum).

The feasibility of exploiting non-gridded bacterial artificial chromosome (BAC) libraries and some major factors affecting the efficiency of handling such libraries were studied in hexaploid wheat. Even for a bacterial culture containing only 55% recombinants, some 2000 BAC clones with inserts ranging from 45 to 245 kb could be pooled. The pooled BAC clones could be amplified by culturing for up to 6 h without losing any target clones. These results imply that even for hexaploid wheat, which has an extremely large genome, some 250 pools are sufficient for a BAC library that should satisfy many research objectives. This non-gridded strategy would dramatically reduce the cost and make robotic equipment non-essential in exploiting BAC technology. To construct a representative library and to minimise clone competition, thawing and re-freezing ligation mixtures and bacterial cultures should be avoided in BAC library construction and application.

Blotting, Southern↗

Aquatic macrophytes as feeding site for small fishes in the Rosana Reservoir, Paranapanema River, southeastern Brazil.

In the present investigation we studied the feeding habits of the fishes associated with aquatic macrophytes in the Rosana Reservoir, southeastern Brazil. Twenty fish species were collected during four field trips, regularly distributed across the dry and wet seasons. Focal snorkeling observations of the fishes were made over a total of six hours. Nine species were present in abundances of more than 1% and, therefore, had their feeding habits analyzed. Hemigrammus marginatus, Roeboides paranensis, Hyphessobrycon eques, Astyanax altiparanae, Serrasalmus spilopleura, and Bryconamericus stramineus were predominantly invertivores, with predominance of aquatic insects (Diptera, Ephemeroptera, and Trichoptera immatures) among their food items. The predominantly algivores were Apareiodon affinis, Serrapinnus notomelas, and Satanoperca pappaterra, with high frequency of filamentous blue-green algae, diatoms, clorophyts, and periderm. The different microhabitat exploitation plus diet composition suggests partitioning of resources and absence of food competition among the most representative fish species in the studied community, indicating the importance of the naturalistic approach to fish ecology studies.

Animals↗

The politics of managed competition: public abuse of the private interest.

The doctrine of managed competition in health care sought to achieve the social goals of access and efficiency using market incentives and consumer choice rather than governmental regulation and public administration. In retrospect, it demanded too much from both the public and the private sectors. Rather than develop choice-supporting rules and institutions, the public sector has promoted process regulation and benefit mandates. The private health insurance sector has pursued short-term profitability rather than cooperate in the development of fair competition and informed consumer choice. Purchasers have subsidized inefficient insurance designs in order to exploit tax and regulatory loopholes and to retain an image of corporate paternalism. America's health care system suffers from the public abuse of private interests and the private abuse of the public interest.

Consumer Behavior↗

Social motives and strategic misrepresentation in social decision making.

In 4 experiments, the authors studied the influence of social motives on deception and strategic misrepresentation. In a newly developed information provision game, individuals faced a decision maker whose decision would affect both own and other's outcomes. By withholding information or by giving (in)accurate information about payoffs, participants could try to influence other's decision making. Less accurate and more inaccurate information was given when the decision maker was competitive rather than cooperative (Experiment 1), especially when participants had a prosocial rather than selfish value orientation (Experiments 3 and 4). Accurate information was withheld because of fear of exploitation and greed, and inaccurate information was given because of greed (Experiment 2). Finally, participants engaged in strategic misrepresentation that may trick competitive others into damaging their own and increasing the participant's outcomes.

Adult↗

A nutritive view on the host-pathogen interplay.

The interaction between pathogenic microbes and their host is determined by survival strategies on both sides, including competition for essential nutrients. During evolution, pathogenic microbes developed ways to access certain nutrients from the host, which, by contrast, can be exploited by the host for defence by restricting the availability of these nutrients. In this article, we review ecological aspects of the host-pathogen relationship and describe examples for competitive nutrient usage. We also discuss the beneficial probiotic microbes of the mammalian gut, which influence their environment including inflammatory host responses, and how they might be supported by prebiotic diets.

Amino Acids↗

Brewing controversies: Darwinian perspective on the adaptive and maladaptive effects of caffeine and ethanol as dietary autonomic modulators.

Ethanol and caffeine are two of the oldest human drugs. Their pervasive integration into the modern human diet may reflect behavioral attempts to correct maladaptations induced by evolutionary displacement of the autonomic system. The dietary adoption of caffeine may parallel the emergence of cognition as an independent basis of competition. Enhancement of the cognitive ability to gather and process information likely evolved as a valuable adjunct to physical behavior in prehistoric fight-or-flight encounters. Caffeine effectively exploits this pre-existing association between adrenergic activity and cognitive readiness, leading to its use in the modern environment where success in competition increasingly depends on cognitive, rather than physical, prowess. Ethanol may have emerged as a dietary means to buffer the maladaptive chronic sympathetic activation and fear response associated with stressful lifestyles and the social phobias associated with the dissolution of kin networks. We explore the health implications of ethanol and caffeine use, with particular attention to their acute and chronic effects on the autonomic axis. The putative protective effects of ethanol in surviving major trauma or reducing inflammation and heart disease may relate to tempering the behavioral and cardiovascular consequences of catastrophic or chronic sympathetic activation. Acute or chronic abuse of ethanol manifests paradoxical pro-adrenergic effects such as tremors and insomnia that may partly represent compensatory responses. Compensatory remodeling may also explain why confirmation of detrimental effects related to caffeine-induced sympathetic activation has proven elusive; indeed, paradoxical pro-vagal benefits may eventually be recognized. Ethanol and caffeine are potential agents that may beneficially expand the dynamic range of the autonomic system. In an environment where the Darwinian value of knowledge has increasingly supplanted that of physical traits, the consumption of caffeine and alcohol may represent both a cause and an effect of modern human evolution.

Adaptation, Physiological↗

Spatially discrete metapopulation models with directional dispersal.

We use an age-structured discrete-time metapopulation model linking two sub-populations through larval transport and directed movements of adults to study the implications of linkages among subpopulations for the stability and resilience of exploited species. Our two-habitat model, a generalization of Fogarty's inshore-offshore lobster population model, includes isolated habitats under compensatory (monotone) or overcompensatory (oscillatory) dynamics [M.J. Fogarty, Implications of migration and larval interchange in American lobster (Homarus americanus) stocks: spatial structure and resilience, in: G.S. Jamieson, A. Campbell (Eds.), Proc. of North Pacific Symposium on Invertebrate Stock Assessment and Management, Can. Spec. Publ. Fish. Aquat. Sci. 125 (1998) 273]. Pre-migration local dynamics are selected from general classes of functions that capture the effects of competition for resources via contest (compensatory) and scramble (overcompensatory) intraspecific competitions. We explore the implications of these mechanisms on the long-term survival of exploited species. In particular, we use threshold parameters R(d)1 for Habitat 1 and R(d)2 for Habitat 2 together with precise mathematical definitions to prove that species persistence is possible at high levels of fishing in one habitat and low to moderate levels of fishing in the other. Our results support Fogarty's conclusion that conservative management of larval source populations could contribute to the resilience of exploited species.

Animal Migration↗

New aspects of image distribution and workflow in radiology.

The progressive use of digital image-generating devices and digital communication technology in clinical and practice environments implies changes in radiological workflow and asks for adequate quality assurance in the whole process of radiology report preparation. This improvement potential has to be rigorously reinvestigated with regard to up-to-date procedures and the full exploitation of supporting technologies like linguistic analysis, help desk and trouble ticket systems, competitive allocation algorithms, time-and-event monitoring, and intelligent agents. These approaches are to be evaluated in combination with business process analysis and shall help to reduce turnaround times for radiology reports while maintaining or even increasing quality-assurance levels.

Germany↗

Human natural yeast killer toxin-like candidacidal antibodies.

A murine mAb (mAbKT4, IgG1) that neutralized in vitro the anti-Candida activity of a killer toxic (KT) from the yeast Pichia anomala acted as an idiotypic (Id) vaccine in eliciting anti-Id Abs with toxin-like activity (KT-IdAb) in a rat vaginitis model. In this study, we demonstrate that intravaginal or intragastric inoculations of Candida albicans bearing a receptor for the toxin was able to recall KT-IdAb production in the vagina of the animals primarily immunized with mAbKT4 and also to elicit by themselves an Ab that functionally mimicked the KT (KTAb). Anti-Id-like, KT-like Abs were also consistently found in the vaginal fluid of human vaginitis patients who were infected by Candida but who had never been exposed to the Id vaccine. These Abs were as candidacidal in vitro as those raised in rat vagina by the Id vaccination, and, likewise, their cytocidal effect was totally neutralized by previous reaction with mAbKT4. Importantly, they were also able to confer a significant anticandidal protection in the rat vaginitis model, comparable to that achievable by KT-IdAb passively transferred to naive rats from Id-vaccinated animals. Thus, candidacidal Abs representing the internal image of a yeast KT are part of the Ab repertoire that follows infection or immunization with Candida. It is speculated that the host's immune system response may exploit the KT receptor of microbial pathogens to produce microbicidal Abs, possibly mirroring competition events among microorganisms in natural habitats.

Administration, Intravaginal↗

Exploitative and hierarchical antagonism in a cooperative bacterium.

Social organisms that cooperate with some members of their own species, such as close relatives, may fail to cooperate with other genotypes of the same species. Such noncooperation may take the form of outright antagonism or social exploitation. Myxococcus xanthus is a highly social prokaryote that cooperatively develops into spore-bearing, multicellular fruiting bodies in response to starvation. Here we have characterized the nature of social interactions among nine developmentally proficient strains of M. xanthus isolated from spatially distant locations. Strains were competed against one another in all possible pairwise combinations during starvation-induced development. In most pairings, at least one competitor exhibited strong antagonism toward its partner and a majority of mixes showed bidirectional antagonism that decreased total spore production, even to the point of driving whole populations to extinction. Differential response to mixing was the primary determinant of competitive superiority rather than the sporulation efficiencies of unmixed populations. In some competitive pairings, the dominant partner sporulated more efficiently in mixed populations than in clonal isolation. This finding represents a novel form of exploitation in bacteria carried out by socially competent genotypes and is the first documentation of social exploitation among natural bacterial isolates. Patterns of antagonistic superiority among these strains form a highly linear dominance hierarchy. At least some competition pairs construct chimeric, rather than segregated, fruiting bodies. The cooperative prokaryote M. xanthus has diverged into a large number of distinct social types that cooperate with clone-mates but exhibit intense antagonism toward distinct social types of the same species. Most lengthy migration events in nature may thus result in strong antagonism between migratory and resident populations, and this antagonism may have large effects on local population sizes and dynamics. Intense mutual antagonism appears to be more prevalent in this prokaryotic social species than has been observed in the eukaryotic social slime mold Dictyostelium discoideum, which also exhibits multicellular development. The finding of several cases of facultative social exploitation among these natural isolates suggests that such exploitation may occur frequently in nature in many prokaryotes with cooperative traits.

Animals↗

Effects of selective contracting on hospital efficiency, costs and accessibility.

This paper exploits a natural experiment in the state of California, to show that pro-competitive healthcare policy may have unintended long-term liabilities unless the system as a whole is carefully designed to preserve access to care for the poor. California's Medicaid Reform Act of 1982 increased competition among hospitals in urban areas, with legislation which allowed direction of patients to more efficient providers via selective contracting. This slowed the average rate of hospital cost inflation, and saved the state billions of dollars. The substantial short-term savings have been documented in empirical research, but little attention has been paid to the longer-term effects of the reforms. We find that Medicaid contracts were awarded to more efficient hospitals. The distributional effects post-reform resulted in efficiency gains for most hospitals, but costs escalated for over half of the public hospitals in the sample, as their uncompensated care burdens rose. Public hospitals continued to fail during the period, leaving over half of California's counties without a county hospital by 1990. Because public hospitals provide the vast majority of healthcare for the poor in California, there is reason for concern about erosion of their access to care as an unintended outcome of pro-competitive reforms.

California↗

Off-patent drugs in Italy. A short-sighted view?

The new Italian policy to off-patent products, although similar to recent reforms in other European countries, seems to use some interesting instruments and has achieved significant results in the past 2 years. In particular, the prices of branded products have been reduced for all active ingredients where a generic version is available. However, this strategy may raise some problems in the longer term. Our analysis identified two open issues that might limit the long-term sustainability of the present scheme: the limited diffusion of generics and the reallocation of demand. The first stems from the new regulatory schemes which equate generics to branded off-patent products and exploit their presence only to cut prices. The second is favored by tough price competition that induces large companies to divert demand towards more profitable "me-too drugs." Solutions exist, but they are not easy to apply because so many difficulties arise when certain aspects of a long-standing equilibrium are modified.

Drugs, Generic↗

FRET-based analysis of TRPC subunit stoichiometry.

By analogy to other cation channel subunits with six transmembrane-spanning domains, the seven members of the "classical" or "canonical" transient receptor potential channels (TRPC) family are believed to assemble into homo- or heterotetrameric complexes. These complexes have been verified by classical methods such as coimmunoprecipitation, crosslinking analysis or functional assays applying dominant negative pore mutants. More recently, fluorescence resonance energy transfer (FRET)-a measure for the close proximity of fluorescent molecules-has become instrumental in monitoring protein assembly in living cells. Here we demonstrate further possibilities and verification procedures of the FRET technology to test the assembly of ion channel subunits. Temporally and spatially resolved FRET imaging demonstrates an early assembly of TRPC subunits in the endoplasmic reticulum and the Golgi apparatus. Confocal FRET imaging verifies FRET signals over the plasma membrane at high spatial resolution. Taking advantage of the quantitative analysis of digital video imaging, we demonstrate that FRET between TRPC subunits is only poorly concentration-dependent. Moreover, a correlation between the efficiency of energy transfer and the molar ratio of the FRET donor to the acceptor was exploited to verify the tetrameric stoichiometry of TRPC complexes. Finally, we introduce a competition-FRET assay to test the ability of wild-type TRPC subunits to recruit fluorescent TRPC subunits into separate channel complexes.

Calcium Channels↗

Density- and frequency-dependent selection at the Mdh-2 locus in Drosophila pseudoobscura.

We have studied differences in the number of Drosophila pseudoobscura produced in a culture when the flies differ with respect to two alleles (F and S) at the Mdh-2 locus, which codes for a malate dehydrogenase enzyme. The studies were done at low and at high density in two- and three-genotype combinations (S/S, F/F and S/F), with one-genotype cultures as controls.--Density affects the fitness of the Mdh-2 genotypes. Different genotypes are differently affected, and the genotype of the competitors also makes a difference on the fitness of a given genotype. When three genotypes are present in a culture, particularly at high density, intergenotypic competition is less intense than intragenotypic competition at several frequency combinations. That is, there is "overcompensation": the three genotypes together exploit the environmental resources better than one genotype alone.--The fitness of the genotypes is frequency dependent in both two-genotype and three-genotype combinations. An inverse relationship between frequency and fitness is observed at high density. This may lead to a stable polymorphism, because the fitness of a genotype increases as its frequency decreases.--Forty independent strains, sampled from a natural population, were used in the experiments. This ensures that more than 95% of the variation present in the genome in the natural population is also present in the experimental cultures. It also ensures that the genetic background on the Mdh-2 alleles is randomized in the same way as it is in nature. However, the possibility remains that Mdh-2 alleles in nature are nonrandomly associated with alleles at closely linked loci. If linkage disequilibrium is present in the experiments because it exists in nature, then the observed effects (such as frequency-dependent selection) would affect the Mdh-2 locus in nature as well.

Alleles↗