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[Sperm flagella and cilia with pathologic motility and ultrastructure].

Motility of sperm flagella as well as of cilia is mechanically based on the principle of 9 + 2-tubules. It functions essentially by coordinated action between microtubules and the adenosine-triphosphatase dynein and was already present at the beginning of the evolution of the eucaryotes. Experimentally induced mutations in algae have resulted in numerous variations of the flagellar 9 + 2-structure. A mutation of this kind is also found in man, as immotile cilia syndrome (ICS) where anomalies in spermatozoa and in cilia (e.g. of the respiratory tract) are observed. Clinical manifestations of the syndrome have long been known (chronic bronchitis, bronchiectasis, sinusitis and male sterility). In addition, half of the patients exhibit situs inversus viscerum, known as Kartagener's syndrome, a subgroup of ICS. Electron microscopy was used to investigate sperm flagella with reduced motility from 9 patients (one with ICS) with primary infertility. Cilia of the respiratory tract from 7 patients (several with ICS) with chronic bronchial problems were analyzed for motility (using video techniques) and ultrastructure. Reduced motility or immotility of spermatozoa and immotile or dyskinetic cilia were always accompanied by ultrastructural anomalies. In spermatozoa, lack of dynein arms, 9 + 0-configuration and extratubuli were most frequently observed. The fibrous sheath was always asymmetrical. Structural ciliary defects resulted in non-parallel arrangements, electron dense matrix substance, extratubuli and lack of radial spokes. In one case, ciliary microtubuli were found in microvilli. In two patients, cilia as well as spermatozoa were analyzed. In the first, immotile spermatozoa without dynein arms and structurally normal cilia were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Effects of cytosine-arabinofuranoside on the development of reptilian embryos (Lacerta viridis, Laur. and Anguis fragilis, L.)].

Administered into the yolk sac of eggs of Lacerta viridis as a single dose of 17 to 40 micrograms, cytosine-arabinoside (Ara-C) was compatible with survival of the embryo, from the sixth day of incubation, for at least 20 to 25 days. The LD50 was 40 to 50 micrograms per egg. Doses of 20 to 40 micrograms of Ara-C introduced in the yolk sac of eggs of the slow-worm (Anguis fragilis) cultured in vitro, at stages of the allantoid bud of 0,5 mm to 2,5 mm long, killed the embryo in 4 to 8 days (possibly due to alterations of capillary blood vessels of allantois and area vasculosa). In the two species, these doses caused cytotoxic effects on embryonic proliferating tissues, growth inhibition and a variety of developmental defects. In young embryos of Anguis fragilis, similar doses of 20 to 40 micrograms of Ara-C caused, in 2 to 4 days, death of many cells in the anlagen of growing organs: neural tube, sensory organs, bronchi, mesoderm of the limb bud, subcutaneous mesenchyme, anlage of dorsal skeletal structures, etc.; followed by growth inhibition and malformations. On the other hand, in the limb bud, the apical ridge was less retrogressed than in control embryos; the limb buds showed slightly better development in treated embryos than in controls, but, Ara-C induced severe damage in their mesoderm. In all embryos of Lacerta viridis, treated at the stage of 6 days or of 10 days of incubation by doses of 20 to 40 micrograms of Ara-C and killed 15 to 35 days later, there was a general reduction of size and of weight and external and internal malformations, more or less severe, were present: modifications of the form of the head, shortening of the lower jaw, labial clefts, microphthalmia, micromelia and other limbs defects, developmental defects of the tail. In some embryos, the only external defects observed were missing fingers and toes; in three of these embryos, the same digits were missing in the four limbs. Modifications of limb morphogenesis induced by Ara-C are compared to structural modifications of the limbs of snake-like Reptilia, and the mechanisms involved in the two series are discussed. These results emphasize the interest of the use of drugs interfering with DNA synthesis, in the field of teratology and in the experimental study of regressive evolution.

Animals↗

Introduction to the neurophysiological study of sleep: central regulation of skeletal and ocular activities.

This paper investigates the historical evolution of the major experimental results in the field of the central mechanisms underlying the skeletal motor activities and the eye movements during sleep. First, the neurophysiological background of paradoxical sleep atonia and paradoxical sleep without atonia was examined. Then, the analysis was directed to the eye movements in humans and animals, their central command processes and their correlative electrophysiological activities: ponto-geniculo-occipital waves and eye movement potentials.

Animals↗

The interaction of class B G protein-coupled receptors with their hormones.

In common with many G protein-coupled receptors, dysfunction in members of the Class B or glucagon-like receptors can elicit a wide spectrum of disease related activities. Consequently, they are potential targets in many different areas of pharmacological research. Unlike the class A or rhodopsin-like receptors, for which at least some structural similarity to bacteriorhodopsin has been detected, absolutely no structural information is available for the Class B G protein-coupled receptors. We present a computational study that exploits the experimental work performed by evolution to indicate residues that are potentially involved in ligand binding in the Class B G protein-coupled receptors. We perform an analysis of mutations that occurred in a correlated fashion between the receptors and their peptidic ligands. The inference that the residues detected in this manner are involved in a direct interaction between the receptor and the ligand is in good agreement with the mutation studies that have already been published.

Amino Acid Sequence↗

Changes in the basic experimental parameters of capillary gas chromatography in the 20th century.

Studies of qualitative changes in capillary gas chromatography are of significant practical and scientific interest. This paper analyzes the evolution of the most important experimental chromatographic parameters over the last three decades and is based on the use of a new approach to scientometrical research that is referred to as applied scientometry. One essential feature of this approach is that it looks at the entire contents of each paper rather than only taking account its title, abstract. and references (as is typical for conventional scientometry). In this paper, we monitor how the most important chromatographic parameters, such as column length and diameter, layer thickness, stationary liquid phases, separation temperature mode. etc., have been evolving over the period 1970-2000. We used data from the following journals: Chromatographia, Journal of Chromatography, and Journal of High Resolution Chromatography and Chromatography Communications.

Chromatography, Gas↗

Functional characterization of left ventricular segmental responses during the initial 24 h and 1 wk after experimental canine myocardial infarction.

Characterization of the temporal evolution of resting segmental function and inotropic reserve after coronary occlusion may be important in evaluating attempts to salvage ischemic but non-necrotic myocardium. Accordingly, we chronically implanted up to six pairs of pulse-transit piezoelectric crystals in the left ventricular myocardium of dogs to measure segmental wall thickness. Segments were separated into groups according to the loss of net systolic thickening (NET) at 5 min postocclusion of the left anterior descending coronary artery in awake, unsedated dogs. Group 1 included segments with NET values of 67--100+ (percent control); group 2 between 67 and 0; and group 3 less than 0 (paradoxical motion). 5 min after coronary occlusion, group 1 NET was 92 +/- 5% (SEM) although significant decreases occurred in NET in group 2 (36 +/- 4%) and group 3 segments (-33 +/- 5%). Between 5 min and 24 h after coronary occlusion, no further significant changes occurred in NET in groups 1, 2, and 3 crystals. Some segments underwent further functional deterioration between 24 h and 1 wk after left anterior descending coronary artery occlusion, although no overall change occurred in segments with mild to moderate ischemic dysfunction. Segments with NET less than 0 at 24 h, on the other hand, exhibited a reduction in aneurysmal bulging between 24 h and 1 wk from -41 +/- 10 to -23 +/- 11% (n = 12, P = 0.02). Inotropic reserve was assessed with postextrasystolic potentiation (PESP) in 14 dogs, and with infusions of dopamine (11 dogs), and isoproterenol (13 dogs). PESP was the most potent intervention and produced a significant augmentation in NET in group 2 crystals at 1, 2, 4, 6,8, and 24 h after coronary occlusion but only at 1 and 2 h in NET in group 3 crystals. Thus, following experimental coronary occlusion, the evolution of ischemic segmental dysfunction is dynamic and variable. A significant degree of inotropic reserve, as assessed by PESP, dopamine, and isoproterenol, exists in segments with moderate ischemic dysfunction for 24 h but for only 2 h after coronary occlusion in those segments with the most severe ischemic dysfunction. In addition, at least some segmental sites with mild to moderate ischemic dysfunction at 24 h deteriorate further between 24 h and 1 wk after experimental coronary occlusion.

Animals↗

Time-resolved infrared absorption studies of the solvent-dependent vibrational relaxation dynamics of chlorine dioxide.

We report a series of time-resolved infrared absorption studies on chlorine dioxide (OClO) dissolved in H2O, D2O, and acetonitrile. Following the photoexcitation at 401 nm, the evolution in optical density for frequencies corresponding to asymmetric stretch of OClO is measured with a time resolution of 120+/-50 fs. The experimentally determined optical-density evolution is compared with theoretical models of OClO vibrational relaxation derived from collisional models as well as classical molecular-dynamics (MD) studies. The vibrational relaxation rates in D2O are reduced by a factor of 3 relative to H2O consistent with the predictions of MD. This difference reflects modification of the frequency-dependent solvent-solute coupling accompanying isotopic substitution of the solvent. Also, the geminate-recombination quantum yield for the primary photofragments resulting in the reformation of ground-state OClO is reduced in D2O relative to H2O. It is proposed that this reduction reflects enhancement of the dissociation rate accompanying vibrational excitation along the asymmetric-stretch coordinate. In contrast to H2O and D2O, the vibrational-relaxation dynamics in acetonitrile are not well described by the theoretical models. Reproduction of the optical-density evolution in acetonitrile requires significant modification of the frequency-dependent solvent-solute coupling derived from MD. It is proposed that this modification reflects vibrational-energy transfer from the asymmetric stretch of OClO to the methyl rock of acetonitrile. In total, the results presented here provide a detailed description of the solvent-dependent geminate-recombination and vibrational-relaxation dynamics of OClO in solution.

Journal Article↗

Sexual conflict selects for male and female reproductive characters.

BACKGROUND: Strict genetic monogamy leads to sexual harmony because any trait that decreases the fitness of one sex also decreases the fitness of the other. Any deviation from monogamy increases the potential for sexual conflict. Conflict is further enhanced by sperm competition, and given the ubiquity of this phenomenon, sexual conflict is rife. In support of theory, experimentally enforced monogamy leads to the evolution of sexual benevolence. In contrast, with multiple mating, males evolve traits causing massive female fitness reductions when female evolution is restrained. Theory also predicts increased investment in spermatogenesis when sperm competition risk is high. While this supposition has correlational support, cause and effect has yet to be firmly established. RESULTS: By enforcing monogamy or polyandry in yellow-dung-fly lines, we have shown experimentally that males from polyandrous treatments evolved larger testes. Furthermore, females from this treatment evolved larger accessory sex glands. These glands produce a spermicidal secretion, so larger glands could increase female ability to influence paternity. Using molecular techniques, we have shown that, consistent with this idea, males' success as second mates is reduced in females from the polyandrous treatment. Nevertheless, males from polyandrous lines achieve higher paternity during sperm competition, and this finding further supports the testis evolution patterns. CONCLUSIONS: These results provide direct experimental support for macroevolutionary patterns of testis size evolution. Furthermore, we have shown that sperm competition selects for traits likely to be important in sexual conflicts over paternity, a result only previously demonstrated in Drosophila melanogaster.

Animals↗

The production of de novo folded proteins by a stepwise chain elongation: a model for prebiotic chemical evolution of macromolecular sequences.

We describe an experimental procedure to mimic the formation of long (over 40 residues) co-oligopetide sequences in many identical copies which may have occurred in the prebiotic molecular evolution. The basic hypothesis is that chain formation is based on the stepwise fragment condensation of randomly generated short oligopeptides, whereby the elongation takes place under the contingent environmental constraints (solubility, pH, salinity), which eliminate most of the products, and thus determine the selection towards one particular small set of chains. The present work aims at verifying the validity of this scheme. In order to do so, we utilize a classic synthetic procedure based on the Merrifield solid-phase synthesis of peptides for the synthesis of randomly produced peptides as well as for their stepwise fragment condensation. Thus, starting from a library of peptides with n=10, the first condensation step produces a library of 16 peptides with 20 residues each (n=20), of which only four remain water-soluble and, therefore, capable to undergo the next fragment condensation step. This gives rise to 16 peptides with n=30, out of which twelve precipitate out under the chosen pH and buffer conditions and are eliminated. Finally, a 44-residue-long water-soluble de novo protein is obtained. This has no homologies or similarities with extant proteins, and, based on circular dichroism (CD), it assumes a stable three-dimensional folding. In agreement with CD data, molecular-modelling simulations suggest an helical fold for the protein with poor, if any, structural homology with known proteins. The implication of this procedure as a general mechanism for the etiology of de novo macromolecular sequences and globular proteins in the origin of life is briefly discussed.

Amino Acid Sequence↗

Punctuated equilibrium in software evolution.

An approach based on the paradigm of self-organized criticality is proposed for experimental investigation and theoretical modeling of software evolution. The dynamics of modifications is studied for three free, open source programs MOZILLA, FREE-BSD, and EMACS using the data from version control systems. Scaling laws typical for self-organized criticality found. A model of software evolution presenting the natural selection principle is proposed. Results of numerical and analytical investigation of the model are presented. They are in good agreement with data collected for real-world software.

Journal Article↗

The adaptive evolution database (TAED).

BACKGROUND: The Master Catalog is a collection of evolutionary families, including multiple sequence alignments, phylogenetic trees and reconstructed ancestral sequences, for all protein-sequence modules encoded by genes in GenBank. It can therefore support large-scale genomic surveys, of which we present here The Adaptive Evolution Database (TAED). In TAED, potential examples of positive adaptation are identified by high values for the normalized ratio of nonsynonymous to synonymous nucleotide substitution rates (KA/KS values) on branches of an evolutionary tree between nodes representing reconstructed ancestral sequences. RESULTS: Evolutionary trees and reconstructed ancestral sequences were extracted from the Master Catalog for every subtree containing proteins from the Chordata only or the Embryophyta only. Branches with high KA/KS values were identified. These represent candidate episodes in the history of the protein family when the protein may have undergone positive selection, where the mutant form conferred more fitness than the ancestral form. Such episodes are frequently associated with change in function. An unexpectedly large number of families (between 10% and 20% of those families examined) were found to have at least one branch with high KA/KS values above arbitrarily chosen cut-offs (1 and 0.6). Most of these survived a robustness test and were collected into TAED. CONCLUSIONS: TAED is a raw resource for bioinformaticists interested in data mining and for experimental evolutionists seeking candidate examples of adaptive evolution for further experimental study. It can be expanded to include other evolutionary information (for example changes in gene regulation or splicing) placed in a phylogenetic perspective.

Adaptation, Physiological↗

Dynamic regulation of the tryptophan operon: a modeling study and comparison with experimental data.

A mathematical model for regulation of the tryptophan operon is presented. This model takes into account repression, feedback enzyme inhibition, and transcriptional attenuation. Special attention is given to model parameter estimation based on experimental data. The model's system of delay differential equations is numerically solved, and the results are compared with experimental data on the temporal evolution of enzyme activity in cultures of Escherichia coli after a nutritional shift (minimal + tryptophan medium to minimal medium). Good agreement is obtained between the numeric simulations and the experimental results for wild-type E. coli, as well as for two different mutant strains.

Anthranilate Synthase↗

Generating solitons by phase engineering of a bose-einstein condensate

Quantum phase engineering is demonstrated with two techniques that allow the spatial phase distribution of a Bose-Einstein condensate (BEC) to be written and read out. A quantum state was designed and produced by optically imprinting a phase pattern onto a BEC of sodium atoms, and matter-wave interferometry with spatially resolved imaging was used to analyze the resultant phase distribution. An appropriate phase imprint created solitons, the first experimental realization of this nonlinear phenomenon in a BEC. The subsequent evolution of these excitations was investigated both experimentally and theoretically.

Journal Article↗

Hemiascomycetous yeasts at the forefront of comparative genomics.

With more than a dozen species fully sequenced, as many as this partially sequenced, and more in development, yeasts are now used to explore the frontlines of comparative genomics of eukaryotes. Innovative procedures have been developed to compare and annotate genomes at various evolutionary distances, to identify short cis-acting regulatory elements, to map duplications, or to align syntenic blocks. Human and plant pathogens, in addition to yeasts that show a variety of interesting physiological properties, are included in this multidimensional comparative survey, which encompasses a very broad evolutionary range. As major steps of the evolutionary history of hemiascomycetous genomes emerge, precise questions on the general mechanisms of their evolution can be addressed, using both experimental and in silico methods.

Adaptation, Physiological↗

Pancreatic duct obstruction in rabbits causes digestive zymogen and lysosomal enzyme colocalization.

The pancreatic duct of anesthetized rabbits was cannulated and, in some animals, flow of pancreatic exocrine secretions was blocked by raising the cannula to a vertical position. Blockage for 3-7 h caused a rapid and significant rise in serum amylase activity and an increase in amylase activity within the pancreas. The concentration of lysosomal enzymes in the pancreas was not altered but they became redistributed among subcellular fractions and, as a result, an increased amount was recovered in the 1,000-g, 15-min pellet, which was enriched in zymogen granules. Immunofluorescence studies indicated that lysosomal enzymes become localized within organelles which, in size and distribution, resemble zymogen granules. They also contain digestive enzyme zymogens. Blockage of pancreatic secretions also caused lysosomal enzyme-containing organelles to become more fragile and subject to in vitro rupture. These changes noted after short-term pancreatic duct obstruction are remarkably similar to those previously noted to occur during the early stages of diet and secretagogue-induced experimental pancreatitis, observations that have suggested that colocalization of digestive enzyme zymogens and lysosomal hydrolases might result in intracellular digestive enzyme activation and be an important early event in the evolution of those forms of experimental acute pancreatitis.

Amylases↗

Cell surface endothelial proteins altered in experimental allergic encephalomyelitis.

Endothelial cells (EC) are increasingly being considered as important participants in the early evolution of inflammatory and immune responses in experimental allergic encephalomyelitis (EAE). We have found that a mouse monoclonal antibody, which reacts with the luminal plasma membrane of central nervous system endothelium, detects an alteration in the blood-brain barrier (BBB) in lesions in Lewis rats with EAE. Anti-endothelial barrier antigen (EBA) reacted with microvessels in normal rat brain and spinal cord. This reaction was abolished in 'EAE' microvessels surrounded by inflammatory cells. In rats that had recovered from one attack most EC reacted with the antibody, indicating that EBA was reexpressed during recovery. However, blood vessels in areas with residual inflammatory lesions were negative. The biochemical changes that lead to this absence of antibody binding and the cells or mediators responsible for producing this change are not yet known. However, anti-EBA should provide a useful tool for exploring molecular mechanisms underlying BBB breakdown.

Animals↗

Dynamics of bacterial phenotype selection in a colonized host.

The population dynamics of Helicobacter pylori during colonization in an infected animal host provide a quantifiable experimental model of in vivo microbial phenotype evolution. Phenotype variability in H. pylori populations can be typed as polymorphic expression of Lewis antigens on their cell surfaces. The high mutational frequency of H. pylori for Lewis expression provides substrate for differential selection by the host. Experimental challenge and successful colonization of mice and gerbils allows tracking of H. pylori phenotype variability from the initial inoculation to the ultimate establishment of a quasispecies. Colonization data provide a quantitative experimental model of phenotype evolution in a relatively large population (>10(4) individuals) over a relatively long evolutionary time scale (>10(3) generations). A mathematical model is developed to interpret the data in terms of the dynamic processes occurring during colonization. The mathematical model distinguishes the roles of selection and mutation; quantifies the effects of initial phenotype diversity, mutational frequency, and selective advantage; and applies generally to phenotype evolution in biological populations.

Animals↗