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Biomaterials and Granulomas

The rapidly expanding use of implants for reconstruction and repair of diseased or traumatized tissues and organs has fueled the search for biomaterials that are stable, nontoxic, and biologically inert. Unfortunately, implants frequently cause acute or chronic inflammation resulting in tissue damage and rejection. Inflammation usually occurs at the biomaterial-tissue interface and reflects surface adsorption of plasma proteins, complement activation, neutrophil and macrophage infiltration, hyperplasia, and release of inflammatory mediators and proteolytic enzymes. If the biomaterial degrades, either spontaneously or due to biologic activity, components can leach into surrounding tissues and may enter the circulation, causing toxic effects systemically and in distant sites. Inflammation of connective tissues in genetically susceptible individuals can potentially activate the immune system and break tolerance to tissue-specific antigens. Experimentally induced animal models of arthritis chronicle the evolution of a chronic, destructive inflammation localized to peripheral joints. Models of autoimmune arthritis involve hyperimmunization with heterologous cartilage components, such as type II collagen or proteoglycan, leading to cross-reactivity with self-cartilage. Other apparently nonimmune models, oil-induced arthritis and pristane arthritis, may reflect granuloma formation and uncontrolled cytokine production. The antigen-induced arthritis model, developed by hyperimmunization with a foreign antigen that is subsequently injected into the joint cavity, is ideal for biocompatibility testing of cationic substances that may be immunogenic. This variety of models offers the means to understand adverse implant-tissue interactions and granuloma formation such that safer biomaterials can be developed in the future.

Journal Article↗

Long-term morphology of the alumina cream experimental epileptogenic focus.

A morpholigical study of an experimental epileptogenic focus in the cat, eighteen months after intracerebral injection of alumina cream, is presented. The most significant histological findings were cortical atrophy and reactive gloisis enclosing the implantedd particles. Subpial sclerosis and evidence of ischaemic neuronal lesions were also found. These findings confirm a close morphological relation between the alumina cream experimental focus obtained after a certain period of evolution and the human epileptogenic focus.

Aluminum↗

RNA based evolutionary optimization.

The notion of an RNA world has been introduced for a prebiotic scenario that is dominated by RNA molecules and their properties, in particular their capabilities to act as templates for reproduction and as catalysts for several cleavage and ligation reactions of polynucleotides and polypeptides. This notion is used here also for simple experimental assays which are well suited to study evolution in the test tube. In molecular evolution experiments fitness is determined in essence by the molecular structures of RNA molecules. Evidence is presented for adaptation to environment in cell-free media. RNA based molecular evolution experiments have led to interesting spin-offs in biotechnology, commonly called 'applied molecular evolution', which make use of Darwinian trial-and-error strategies in order to synthesize new pharmacological compounds and other advanced materials on a biological basis. Error-propagation in RNA replication leads to formation of mutant spectra called 'quasispecies'. An increase in the error rate broadens the mutant spectrum. There exists a sharply defined threshold beyond which heredity breaks down and evolutionary adaptation becomes impossible. Almost all RNA viruses studied so far operate at conditions close to this error threshold. Quasispecies and error thresholds are important for an understanding of RNA virus evolution, and they may help to develop novel antiviral strategies. Evolution of RNA molecules can be studied and interpreted by considering secondary structures. The notion of sequence space introduces a distance between pairs of RNA sequences which is tantamount to counting the minimal number of point mutations required to convert the sequences into each other. The mean sensitivity of RNA secondary structures to mutation depends strongly on the base pairing alphabet: structures from sequences which contain only one base pair (GC or AU are much less stable against mutation than those derived from the natural (AUGC) sequences. Evolutionary optimization of two-letter sequences in thus more difficult than optimization in the world of natural RNA sequences with four bases. This fact might explain the usage of four bases in the genetic language of nature. Finally we study the mapping from RNA sequences into secondary structures and explore the topology of RNA shape space. We find that 'neutral paths' connecting neighbouring sequences with identical structures go very frequently through entire sequence space. Sequences folding into common structures are found everywhere in sequence space.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Physiological↗

Experimental occlusion of the retinal vein.

We used a pars plana approach to experimentally induce retinal vein occlusions (RVO) in cats by the application of electric energy near the veins at the optic nerve head. This experimental model was utilized to study the natural evolution of thrombosis in the cilioretinal veins, which are located near the lamina cribrosa in the cat. We observed that occlusion of the retinal veins occurs by the formation of a thrombus and that with time there is associated endothelial cell proliferation and recanalization in each instance.

Animals↗

Variations in polymer fitness at elevated mutation rates.

Two series expressions were obtained that give the first and second order rates of change in population fitness during competitive replication at elevated mutation rates. At their zero-error limit, the respective power series reduces to the second (Fisher's theorem) and third moments of the fitness distribution. The first series maximized the variation in mean polymer fitness, for a given amount of population covariance. From experimental results reported by Spiegelman's group on evolution in vitro among Q beta RNA variants, it was demonstrated: (i) terms in the (second) series fall-off at a rate broadly equal to the replicase error (epsilon congruent to 10(-4); (ii) the rate of change in mean RNA fitness (polymer formation rate constant) corresponds to the variance in fitness; and (iii) agreement exists between second order rate changes in fitness and the third moment (skewness) regression line, extending over 20 successive replication reactions. The impact of these findings on the standard model of evolution has been discussed.

Biological Evolution↗

Neglected traumatic posterior dislocations of the shoulder: controversies on indications for treatment and new CT scan findings.

We report seven missed traumatic posterior dislocations of the shoulder in six patients. In all seven shoulders, anteroposterior radiographs did not provide the diagnosis in the emergency room after the trauma (epileptic seizure in four patients) and diagnosis was delayed for at least 24 h. The final diagnosis was reached by clinical suspicion and computed tomography (CT) scan of the seven shoulders. Two shoulders were treated by closed reduction and three were treated by closed reduction and percutaneous pinning, although four of these five shoulders presented a defect in the humeral head involving 20%-25% of the articular surface. Two shoulders dislocated for more than 6 months were treated according to Mc Laughlin's technique modified by Hawkins instead of an arthroplasty. At a minimum follow-up of 2 years and 2 months, the functional results according to Hawkins were good in all seven shoulders. There was no recurrence of the dislocation in any patient. Radiographs showed mild degenerative changes. The establishment of indications for treatment, based on the proportion of articular surface of the humeral head involved and the time of evolution of the dislocation, needs more clinical or experimental evidence. In epileptic patients, strict control of medication is of the utmost importance.

Adult↗

Hydrofluoric acid burns: a review.

Hydrofluoric acid (HF) is a highly dangerous substance with a wide range of industrial as well as domestic applications. It is unique in both the severity of the cutaneous burns it may produce, and its potential for systemic and occasionally lethal toxicity. The literature on the treatment of HF injuries is extensive, though occasionally confusing and often contradictory, with no coherent management policy emerging. In this paper we present a comprehensive account of the evolution of therapy, drawing on clinical reports and experimental studies.

Burns, Chemical↗

Development of an inverse method to identify the kinetics of heavy metal release during waste incineration in fluidized bed.

This paper deals with the emission of heavy metals (HM) during the incineration of municipal solid waste in a fluidized bed reactor. This study focused on the development of a general method to identify the kinetics of vaporization of heavy metals from the on-line analysis of exhaust gas. This method is an inverse method, which requires only the time evolution of the HM concentration in exhaust gases (experimental data) and a global bubbling bed model developed for transient conditions at the reactor scale. First, a lab-scale fluidized bed incinerator was set-up to simulate the HM release during the thermal treatment of metal-spiked model wastes. A specific on-line analysis system based on ICP-OES was developed to measure in real time the variation of the relative concentration of HM in exhaust gases. Then, a two-phase flow bubbling bed model was developed and validated to calculate the kinetics of vaporization of HM from its measured concentration time profile in the outlet gas. The technique was first validated with model waste (metal-spiked mineral matrices), thus enabling at each time both solid sampling for measuring the HM vaporization kinetic and on-line analysis for measuring the HM concentration in the outlet gas. The inverse method was then applied to realistic artificial wastes (derived from real wastes) to identify the HM vaporization kinetics from the on-line analysis results.

Aluminum Oxide↗

Use of active consortia of constructed ternary bacterial cultures via mixture design for azo-dye decolorization enhancement.

This first-attempt study used constructed bacterial consortia containing Escherichia coli DH5alpha (a weak decolorizer) and its UV-irradiated mutants (E. coli UVT1 and UV68; strong decolorizers) via equilateral triangle diagram and mixture experimental design to assess color removal during species evolution. The results showed that although strain DH5alpha was not an effective decolorizer, its presence might still played a significant role in affecting optimal color removal capabilities of mixed consortia (e.g., E. coli DH5alpha, UVT1 and UV68) for two model azo dyes; namely, reactive red 22 (RR22) and reactive black 5 (RB5). Contour analysis of ternary systems also clearly showed that decolorization of RR22 and RB5 by DH5alpha-containing active mixed consortia was more effective than mono-cultures of the stronger decolorizer alone (e.g., UVT1). The optimal composition of the mixed consortium (UV68, UVT1, DH5alpha) achieving the highest specific decolorization rate was (13%:58%:29%) and (0%:74%:26%) for decolorization of RR22 and RB5, respectively, with initial total cell density fixed at OD(600)=3.5+/-0.28.

Azo Compounds↗

Headspace solid-phase microextraction method for determining 3-alkyl-2-methoxypyrazines in musts by means of polydimethylsiloxane-divinylbenzene fibres.

A method for determining 2-methoxypyrazine, 3-methyl-, 3-ethyl-, 3-isopropyl-, 3-sec.-butyl- and 3-isobutyl-2-methoxypyrazine in musts is described. It involves headspace solid-phase microextraction (SPME) and determination by capillary gas chromatography using nitrogen-phosphorous detection. Pyrazines were satisfactorily separated under isothermal conditions, and quantification was carried out using 3-isopropyl-2-ethoxypyrazine as the internal standard. Ionic strength, time and temperature were studied in order to make SPME as efficient as possible. The developed method enabled detection limits at the 0.1 ng(-1) levels for some of the analytes. The method was successfully applied to identify and quantify different 3-alkyl-2-methoxypyrazines in experimental musts of Cabernet Sauvignon and Merlot. Their evolution during the ripening was also monitored.

Osmolar Concentration↗

Drosophila in cancer research. An expanding role.

In recent years, Drosophila researchers have developed powerful genetic techniques that allow for the rapid identification and characterization of genes involved in tumor formation and development. The high level of gene and pathway conservation, the similarity of cellular processes and the emerging evidence of functional conservation of tumor suppressors between Drosophila and mammals, argue that studies of tumorigenesis in flies can directly contribute to the understanding of human cancer. In this review, we explore the historical and current roles of Drosophila in cancer research, as well as speculate on the future of Drosophila as a model to investigate cancer-related processes that are currently not well understood.

Animals↗

The concept of preleukemia: clinical and laboratory studies.

Thirty-five years ago, a handful of astute clinical hematologists began to notice that some of their patients with acute nonlymphocytic leukemia had a history of a preceding ill-defined hemopathy. This "preleukemic" hemopathy was increasingly reported anecdotally and through careful retrospective analyses. In more recent prospective studies, this syndrome has been relatively well defined. Indeed, it is widely accepted that there exists a hematopoietic abnormality not recognizable as classical, overt, acute nonlymphocytic leukemia, which, nonetheless, can represent an early phase of leukemia. In this manuscript, the author reviews the original case reports, the initial retrospective studies, and the prospective studies which have clarified some of the clinical and laboratory features of the preleukemic syndrome. The notion that the preleukemic syndrome represents an established neoplastic process will be reviewed with special attention to assessment of clonal hematopoiesis, cytogenetic studies, and clonal evolution. Studies on induced leukemia and preleukemia in experimental animals and humans will be reviewed as will be the semantic issues which have complicated our ability to compare interinstitutional studies. A simple classification scheme of the myelodysplastic syndromes will be presented as will be a framework agenda for future studies on the pathophysiology of these syndromes.

Animals↗

Enzyme immunoassay for myelin basic protein in cerebrospinal fluid.

Myelin basic proteins (MBPs) are a set of proteins making up about 30% of the protein content of the central nervous system myelin. Four human isoforms have been identified. The most abundant is a highly conserved 18.5 kDa polypeptide. For this species, the amino acid sequence homologies between human and monkey or human and chick are 98.2% and 71.1%, respectively. As a consequence, there is a very good immunological cross-reactivity between the mammalian MBP. This protein has been extensively used to induce experimental allergic encephalomyelits (EAE) in numerous animals. The evolution of chronic EAE in animal is similar to that of multiple sclerosis (MS), a demyelinating human pathology, and chronic EAE is considered to be an animal model of MS. In demyelinating pathologies, MBP concentration in the cerebrospinal fluid (CSF) is considered to be a good marker of demyelination. MBP concentration, in biological fluids, is generally determined by radioimmunoassay (RIA). The RIA technique currently used is highly sensitive (0.1-2.5 ng/ml) but has the drawback of requiring the handling of radioactivity and frequent labelling of MBP. So we developed a new enzyme immunoassay (EIA) technique. Our technique has the same sensitivity as RIA, needs only small volumes of CSF (50 microliters) and the enzyme-labelled MBP tracer is stable for at least 12 months.

Alkaline Phosphatase↗

Photoelectron spectroscopy of doubly and singly charged group VIB dimetalate anions: M2O72-, MM'O72-, and M2O7- (M, M' = Cr, Mo, W).

We produced both doubly and singly charged Group VIB dimetalate species-M(2)O(7)(2-), MM'O(7)(2-), and M(2)O(7)(-) (M, M' = Cr, Mo, W)-using two different experimental techniques (electrospray ionization for the doubly charged anions and laser vaporization for the singly charged anions) and investigated their electronic and geometric structures using photoelectron spectroscopy and density functional calculations. Distinct changes in the electronic and geometric structures were observed as a function of the metal and charge state. The electron binding energies of the heteronuclear dianions MM'O(7)(2-) were observed to be roughly the average of those of their homonuclear counterparts (M(2)O(7)(2-) and M'(2)O(7)(2-)). Density functional calculations indicated that W(2)O(7)(2-), W(2)O(7)(-), and W(2)O(7) possess different ground-state structures: the dianion is highly symmetric (D(3d),(1)A(1g)) with a single bridging oxo ligand, the monoanion is a doublet (C(1), (2)A) with two bridging oxo ligands and a radical terminal oxo ligand, whereas the neutral is a singlet (C(1), (1)A) with two bridging oxo ligands and a terminal peroxo ligand. The combined experimental and theoretical study provides insights into the evolution of geometric and electronic structures as a function of charge state. The clusters identified might provide insights into the possible structures of reactive species present in early transition-metal oxide catalysts that are relevant to their reactivity and catalytic function.

Anions↗

Genetic cost of reproductive assurance in a self-fertilizing plant.

The transition from outcrossing to self-fertilization is one of the most common evolutionary trends in plants. Reproductive assurance, where self-fertilization ensures seed production when pollinators and/or potential mates are scarce, is the most long-standing and most widely accepted explanation for the evolution of selfing, but there have been few experimental tests of this hypothesis. Moreover, many apparently adaptive floral mechanisms that ensure the autonomous production of selfed seed might use ovules that would have otherwise been outcrossed. This seed discounting is costly if selfed offspring are less viable than their outcrossed counterparts, as often happens. The fertility benefit of reproductive assurance has never been examined in the light of seed discounting. Here we combine experimental measures of reproductive assurance with marker-gene estimates of self-fertilization, seed discounting and inbreeding depression to show that, during 2 years in 10 Ontario populations of Aquilegia canadensis (Ranunculaceae), reproductive assurance through self-fertilization increases seed production, but this benefit is greatly outweighed by severe seed discounting and inbreeding depression.

Fertility↗

Limited boundaries for extensive horizontal gene transfer among Salmonella pathogens.

Recombination is thought to be rare within Salmonella, as evidenced by absence of gene transfer among SARC strains that represent the broad genetic diversity of the eight primary subspecies of this common facultative intracellular pathogen. We adopted a phylogenetic approach to assess recombination within the mutS gene of 70 SARB strains, a genetically homogeneous population of Salmonella enterica subspecies I strains, which have in common the ability to infect warm-blooded animals. We report here that SARB strains show evidence for widespread recombinational exchange in contrast to results obtained with strains exhibiting species-level genetic variation. Besides extensive allele shuffling, SARB strains showed notably larger recombinagenic patch sizes for mutS (at least approximately 1.1 kb) than previously reported for S. enterica SARC strains. Explaining these experimental dichotomies provides important insight for understanding microbial evolution, because they suggest likely ecologic and genetic barriers that limit extensive gene transfer in the feral setting.

Base Sequence↗

Landmarks in territory partitioning: a strategically stable convention?

A convention is a rule based on arbitrary cues that allows quick resolution of potentially protracted disputes. A familiar example is the Bourgeois strategy, in which the second of two animals to discover a resource yields it to the first, even though it may be stronger than its opponent. Here we develop a game-theoretic model to show that neighbors with imperfect information about one another's fighting abilities can be favored to accept a landmark as the designator of a territory boundary, even when the resulting territory is smaller than the one that would have been won through fighting. Thus, the use of landmarks or other mutually obvious solutions can serve as a convention for territory partitioning. For a distribution of fighting ability with low variance and high skew, there is a remarkably high probability that an animal will accept a smaller territory than it would have won through fighting. The analysis provides a possible explanation for the observed use of landmarks as boundary markers by territorial animals in a variety of taxa, including birds, fish, insects, and mammals. The analysis also suggests why territory boundaries are stable, once established, despite changes in characteristics of the residents or the environment.

Biological Evolution↗

Quantitative modeling and data analysis of SELEX experiments.

SELEX (systematic evolution of ligands by exponential enrichment) is an experimental procedure that allows the extraction, from an initially random pool of DNA, of those oligomers with high affinity for a given DNA-binding protein. We address what is a suitable experimental and computational procedure to infer parameters of transcription factor-DNA interaction from SELEX experiments. To answer this, we use a biophysical model of transcription factor-DNA interactions to quantitatively model SELEX. We show that a standard procedure is unsuitable for obtaining accurate interaction parameters. However, we theoretically show that a modified experiment in which chemical potential is fixed through different rounds of the experiment allows robust generation of an appropriate dataset. Based on our quantitative model, we propose a novel bioinformatic method of data analysis for such a modified experiment and apply it to extract the interaction parameters for a mammalian transcription factor CTF/NFI. From a practical point of view, our method results in a significantly improved false positive/false negative trade-off, as compared to both the standard information theory based method and a widely used empirically formulated procedure.

Animals↗