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PubMed1994

Period dependent selection in continuous culture of viruses in a periodic environment.

Selection in a cellstat culture of two mutant strains of a bacteriophage under periodically fluctuating temperature was analyzed mathematically and numerically. Each of the two viral strains (P1 and P2) was assumed to have a different Arrhenius activation energy for its reproduction reaction. A phase diagram of the final state in the continuous culture was drawn. The most noticeable was that there were both P1-only and P2-only phases of competitive exclusion depending on the period of oscillation and the dilution rate. The period-dependent selection was proved that it was based on the feedback effect via the host-population change. At a short period of oscillation, the strain with larger arithmetic average fitness ultimately dominated in the population; on the other hand, at a long period of oscillation, the strain with larger geometric average fitness ultimately dominated. There was a co-existence phase between the two exclusion phases. The results suggest the feasibility of a method to escape from trapping at local optima on a fitness landscape in evolutionary molecular engineering.

Bacteriophages↗
PubMed1994

Background conductance attributable to spontaneous opening of muscarinic K+ channels in rabbit sino-atrial node cells.

Single myocytes were dissociated from the rabbit sino-atrial node, and the membrane background conductance produced by spontaneous opening of the muscarinic K+ channels was investigated by recording whole-cell and single channel currents in both normal K+ (5.4 mM) and high-K+ (145 mM) external solutions. Increasing external K+ concentration ([K+]o) from 5.4 to 145 mM induced a large inward shift of the whole-cell current accompanied by considerable current fluctuations at -50 mV. The high-K(+)-induced current was both K+ selective and voltage dependent, which was examined by varying [K+]o. This current was almost completely suppressed by 1-5 mM Ba2+ or 2-10 mM Cs+ and it was partly blocked by 10 microM atropine. In high-K+ (145 mM) solution, 20 nM acetylcholine (ACh) further increased the K+ conductance as well as the current noise. The power density spectrum of the noise was fitted with a sum of two Lorentzian functions. The corner frequencies of both the slow (approximately 5 Hz) and fast (approximately 120 Hz) components were comparable between the noise before and during the ACh application. Internal dialysis with a non-hydrolysable derivative of ATP, 5'-adenylylimido-diphosphate (AMP-PNP) or Mg(2+)-free solution markedly decreased both the amplitude and fluctuations of the high-K(+)-induced current. The relation between the variance of the current fluctuations and the mean current amplitude was linear in every experiment using dialysis of AMP-PNP or Mg(2+)-free internal solution, or using superfusion of ACh. The slopes of these relations gave comparable single channel current amplitudes of -0.7 pA at -50 mV. These results indicate that the spontaneous opening of the muscarinic K+ channels is largely responsible for the high-K(+)-induced current. In the high-K+ solution, the variance-mean relation at -50 mV showed that the muscarinic K+ channel provides an inward current of 3.12 +/- 2.13 pA pF-1 (n = 23), which was about 60% of the total inward background current. In the normal K+ solution, the variance-mean relation at -50 mV indicated that an outward current of 6.0 +/- 2.0 pA (0.33 +/- 0.28 pA pF-1, n = 8) was provided by the K+ channel. The single channel current amplitude was estimated to be 0.06 +/- 0.02 pA (n = 9). Cell-attached recordings in the absence of ACh demonstrated sporadic and brief openings of channels identical to the ACh-induced channels. The power density spectra of the single channel currents exhibited kinetic properties comparable with those of the whole-cell currents.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗
PubMed1992

A stochastic evolutionary model of molecular sequences.

A stochastic evolutionary model of molecular sequences is proposed. The basic forces in evolution are supposed to be mutation and selection. The concept is somewhat similar to Kauffman-Levin's concept of adaptive walks and corresponding analytical expressions have been developed. The selective force is divided into two parts: a slowly-varying part and a rapidly-changing fluctuation. The latter influences the distribution of sequences and results in an equation of motion along the flow line. The former plays a more important role in the emergence of evolutionary order. It is demonstrated that the asymmetry of selective forces would lead to a definite order of the system.

Animals↗
PubMed1991

Stable ornithine decarboxylase in a rat hepatoma cell line selected for resistance to alpha-difluoromethylornithine.

Ornithine decarboxylase (ODC) is extremely unstable in mammalian cells. This unusual characteristic facilitates rapid fluctuations in the activity of this enzyme in response to variations in its biosynthesis. Unfortunately, very little is known about the mechanism or regulation of this ODC-specific proteolytic pathway. This study describes the production and characterization of a variant of the rat hepatoma HTC cell line that is strikingly deficient in this pathway. This cell variant was induced by selection for growth in stepwise increasing concentrations (up to 10 mM) of the irreversible ODC inhibitor, alpha-difluoromethylornithine (DFMO). Resistance to this inhibitor appears to result from a combination of elevated (10X) ODC biosynthesis and inhibited degradation, producing greater than a 2000-fold increase in the level of ODC protein. In these variant cells (DH23b) inhibition of protein synthesis by cycloheximide did not result in rapid loss of enzyme activity or ODC protein determined by radioimmunoassay. Pulse-chase studies with [35S]methionine confirmed that this enzyme was not preferentially degraded, even when spermidine was added to the media. ODC purified from the variant cells was found to be identical to the control cell enzyme in size, isoelectric point, substrate binding kinetics, and sensitivity to the inhibitor DFMO. Also, as in the control cells, a major fraction of the ODC molecules extracted from DH23b cells was shown to be phosphorylated on a serine residue. The inability to detect physical or kinetic differences between the parent and the variant cell ODC suggests that the unusual stability of ODC in this cell is associated with a defect in a cellular mechanism for ODC-specific degradation.

Animals↗
PubMed2003

Thermoregulation is the pits: use of thermal radiation for retreat site selection by rattlesnakes.

Pitvipers (Viperidae: Crotalinae) possess unique sensory organs, the facial pits, capable of sensing subtle fluctuations in thermal radiation. Prey acquisition has long been regarded as the sole function of the facial pits. However, the ability to sense thermal radiation could also direct thermoregulatory behavior by remotely sensing nearby surface temperatures. Using a series of behavioral arenas of varying spatial complexity and ecological relevance, we surveyed the ability of the western diamondback rattlesnake Crotalus atrox to direct successful thermoregulatory movements with either functional or disabled facial pits. We found that western diamondback rattlesnakes could base thermoregulatory decisions on thermal radiation cues when their pits were functional, but not when blocked. Our results indicate that the facial pit is part of a generalized sense, and suggest thermoregulation as an alternative hypothesis to prey acquisition for the origin of facial pits.

Animals↗
PubMed1982

Loneliness as a function of selected personality variables.

Hypothesized that in a multivariate statistical model, selected personality variables, depression, anxiety, neuroticism, psychoticism, misanthropy, and external locus of control, could positively predict loneliness, and self-esteem and extraversion could negatively predict loneliness scores. Two groups of Ss were studied independently. Ss in Group I were 232 Iranian college students (156 males, 76 females) who were studying in American colleges. Group II consisted of 305 Iranian students (168 males, 137 females) who were studying in Iranian universities. The obtained results, applying multiple regression analysis, confirmed the directions stated in the research hypothesis. However, some of the selected variables did not contribute significantly in the regression equations. Because of fluctuation of the regression coefficients, due to multicolinearity, the data were subjected to factor analysis. Two factors emerged with eigenvalues greater than unity. Loneliness loaded heavily on the first factor, which was identified as negative attribute of personality.

Adult↗
PubMed2006

Biodiversity of the genus Fusarium in saline soil habitats.

Fusarium species assemblage and diversity were investigated in eight different contrasting extreme saline soil habitats of the hot arid desert environment of Bahrain. Saline habitats are located towards the central-southern part of Bahrain and featured by high electrical conductivity, slightly alkaline sandy soil, poor in nutrient sources and water holding capacity and mainly dominated by a salt-tolerant flora. Quantification of data for the recovery of Fusarium species was based on morphological characters and counts by a series of ten fold dilutions plate method and direct soil plating, using two selective media supplemented with different NaCl concentrations. A total of 68 isolates, fluctuated between 1 and 23 per soil sample, were recovered among all habitats mostly at 0 and 5% NaCl concentrations, while no recovery was achieved at 20 and 25%. Grouping of these isolates has resulted in only five species (F. solani, F. oxysporum, F. chlamydosporum, F. equiseti and F. compactum), all of which were previously reported from the arid terrestrial habitats of Bahrain. F. solani was the most predominant species, based on relative density, frequency of occurrence and dominance values, followed by F. oxysporum, a finding consistent with other similar arid Sahara ecosystems. Evaluation of data, supported by analysis of diversity indices and community coefficients, revealed that desert mountain habitat followed by burial mounds habitat were highly homogeneous coupled with maximum species richness, diversity indices and evenness, whereas soil habitats like cliffs, coastal and Al-Lowzy pit were the poorest. Moreover, in vitro tests showed that among other fusaria, F. solani exhibited the highest tolerance to increase NaCl concentrations (25%) and temperature (28.3 mm linear growth at 35 degrees C). At 10% NaCl concentrations, significant reduction in linear growth extensions suppressed all species accompanied by massive thick-walled, drought-resistant chlamydospores formation, indicating that species are mainly thermo-osmotolerant. Fusarium species recoveries were negatively significantly correlated (p < 0.05), with soil edaphic factors including salinity (r = -0.849) and soil moisture (r = -0.711). It is thus, proposed that inter and intra specific variation in the Fusarium species community, in particular soil niche, is an outcome of the influence of bioclimatic, multi-soil-edaphic and biotic factors.

Bahrain↗
PubMed2002

Adenosine A2A antagonism reverses levodopa-induced motor alterations in hemiparkinsonian rats.

To evaluate the possible involvement of adenosine A(2A) receptor-mediated mechanisms in levodopa-induced motor fluctuations, we investigated the effects of CSC (8-(3-chlorostryryl) caffeine), a selective adenosine A(2A) receptor antagonist, on levodopa-induced motor alterations in rats with unilateral 6-OHDA lesion. Acute and chronic administration of CSC was studied to evaluate the possible reversion or prevention of these levodopa effects. In a first set of experiments, rats were treated with levodopa (25 mg/kg with benserazide, twice daily, i.p.) for 22 days and on day 23 CSC (5 mg/kg, i.p.) was administered immediately before levodopa. In a second set of experiments, rats were treated daily for 22 days with levodopa and CSC (5 mg/kg/day, i.p.). The duration of the rotational behavior induced by chronic levodopa decreased after 22 days (P < 0.05). Acute administration of CSC on day 23 reversed levodopa-induced shortening in motor response duration (P < 0.01). Chronic CSC administration did not prevent the shortening in response duration induced by levodopa. Our results demonstrate that the adenosine A(2A) receptor antagonist CSC reverses but does not prevent levodopa-induced motor alterations in parkinsonian rats. These results suggest a role for adenosine A(2A) receptor-mediated mechanisms in the pathophysiology of levodopa-induced motor response complications. These findings suggest that the antagonism of adenosine A(2A) receptors might confer clinical benefit to parkinsonian patients under levodopa therapy suffering from motor complication syndrome.

Animals↗
PubMed1992

Spectral change in heart rate variability in response to mental arithmetic before and after the beta-adrenoceptor blocker, carteolol.

Spectral analysis of heart rate fluctuation was evaluated before and after administration of carteolol, a non-selective beta-adrenoceptor-blocker, to investigate the neural regulatory mechanisms underlying the haemodynamic changes induced by mental stress. Mental stress increased blood pressure and heart rate, with an increased low frequency band, and low frequency/high frequency ratio of the power spectral analysis which are indices of sympathetic activity. Carteolol did not change basal and pre-mental stress measurements of blood pressure, heart rate and spectral density. However, carteolol altered the response to mental stress with a decrease in spectral density of the low frequency band and low frequency/high frequency ratio, and an increase in the high frequency component. These results confirm that mental stress elevates blood pressure by activating the sympathetic nervous system, and suggest that blockade of the beta-adrenoceptor attenuates the pressor response by preventing the autonomic responses to mental stress.

Blood Pressure↗
PubMed1992

Characteristics of juice from palmyrah palm (Borassus) fruit.

The ancestral palmyrah palm (Borassus aethiopum Mart.) produces in the V-Baoulé region of Côte d'Ivoire, sweet and fragrant fruits in large numbers. Most of these spoil in the bush when ripe. The present work was to develop an efficient process for the extraction of juice from these fruits using a pectinase (Pectinex Ultra SP-L) at different concentrations (0.2, 0.4 and 0.6 mg/kg), temperatures (25 degrees C, 35 degrees C and 45 degrees C), and for different times (30, 60, 90 and 120 min). Some physico-chemical characteristics of the juice were determined in order to evaluate the quality of the product. Treatment at 45 degrees C for 120 min at an enzyme concentration of 0.6 mg/kg was selected for highest juice yield (73.14%). The colour parameters L; a; b) fluctuated in intensity between batches of rehydrated material. Juice consistency was found to be stable at 45 degrees C at the enzyme concentrations used. There was a slight variation in pH among the different treatments. In general, the yield and quality of the juice obtained were affected by treatment temperature, enzyme concentration and maceration time.

Beverages↗
PubMed1983

Competitive blocking of apical sodium channels in epithelia.

Apical sodium-selective channels in frog skin, when blocked by amiloride or triamterene, exhibit fluctuations in current, the spectra of which are Lorentzian. These effects have been modeled previously with two-state and three-state models by Lindemann and Van Driessche. A recent observation by Hoshiko and Van Driessche that corner frequencies are lowered by increasing the apical sodium concentration cannot be accounted for by these models. We explore the possibility that sodium (S) and amiloride (A) compete for a site at the mouth of the channel. A new three-state channel model (sodium-occupied, open/unoccupied, open/amiloride-blocked) is analyzed. Its corner frequency is of the form fc = fco [1 + (A/KA)/(1 + S/KS)], consistent with the observed sodium dependence of the corner frequency. The minimum frequency, fco, and the inhibition constants, KA and KS, are expressed in terms of the rate constants of the model. To account for sodium self-inhibition, we postulate that two sodium ions in the channel may result in clogging--a fourth state. The two corner frequencies are calculated; so are the plateau values of the noise power. The noise power shows a maximum as a function of blocker concentration, as observed previously using triamterene. The four-state model predicts the observed suppression by small amounts of blocker of the low-frequency sodium (clogging) noise.

Amiloride↗
PubMed1991

Influence of phloretin and alcohols on barrier defects in the erythrocyte membrane caused by oxidative injury and electroporation.

Oxidative damage by diamide, periodate and oxygen-derived reactive species, but also exposure to electroporation induce in the erythrocyte membrane dynamic, presumably fluctuating, defects having the properties of aqueous holes with definable radii and selectivities. These leaks, which can be quantified by measuring tracer fluxes or rates of colloid-osmotic lysis, are here shown to be inhibited by phloretin and a small number of related phenol compounds (phenolphthalein, hydroxyacetophenones, nitrophenol), while a host of other 'membrane-active' agents is not effective in this respect. I50 values range from about 200 microM for phloretin and phenolphthalein to about 10 mM for 4-nitrophenol. Inhibition by phloretin is reversible, not competitive and not related in its extent to the extent of leakiness. In contrast, the enhancement of transbilayer mobility of amphiphilic lipid probes, which invariably goes along with leak formation of the type described, is not affected by phloretin. Aliphatic alcohols (hexanol, butanol) have an amplifying effect on leaks induced by oxidative damage but do not affect leaks induced by electroporation. The alcohol-amplified leaks maintain the properties of aqueous holes as indicated by a low activation energy of leak fluxes. Since both, inhibition and stimulation of leak fluxes do not go along with appreciable changes of the apparent radii of the aqueous holes, changes in the dynamics (opening and closing) of the defects are proposed to underly the effects of phloretin and alkanols. The membrane lipid domain is likely to be the site of the leaks and of their modulation.

Alcohols↗
PubMed1993

Effect of constraints by threonine on proline containing alpha-helix--a molecular dynamics approach.

Proline plays an important role in the secondary structure of proteins. In the pursuit of understanding its structural role, Proline containing helices with constraints have been studied by employing molecular dynamics (MD) technique. In the present study, the constraint introduced is a threonine residue, whose sidechain has intramolecular hydrogen bond interaction with the backbone oxygen atom. The three systems that have been chosen for characterization are: (1) Ace-(Ala)12-Thr-Pro-(Ala)10-NHMe, (2) Ace-(Ala)13-Pro-Ala-Thr-(Ala)8-NHMe and (3) Ace-(Ala)13-Pro-(Ala)3-Thr-(Ala)6-NHMe. The equilibrium structures and structural transitions have been identified by monitoring the backbone dihedral angles, bend related parameters and the hydrogen bond interactions. The MD averages and root mean square (r.m.s.) fluctuations are compared and discussed. Energy minimization has been carried out on selected MD simulated points in order to analyze the characteristics of different conformations.

Amino Acid Sequence↗
PubMed2002

Testing of the portal hypothesis: analysis of a V32G, F57G, K58G mutant of the fatty acid binding protein of the murine adipocyte.

The portal region of fatty acid binding proteins is hypothesized to function as a dynamic aperture, controlling accessibility of external ligands to the internal fatty acid binding cavity. To test this hypothesis, a triple mutant of the murine FABP4 has been developed (V32G, F57G, K58G, referred to as the portal mutant) that is predicted to constitutively enlarge the opening due to a reduction in the molecular dimensions of the side chains of key portal amino acids. The portal mutant was purified from expressing Escherichia coli, its stability was evaluated, and the thermodynamics and kinetics of ligand binding were compared to that of wild-type protein. Introduction of the three amino acid substitutions caused no significant change in the stability of the protein with a free energy of unfolding of 13.7 kJ/mol as compared to 14.0 kJ/mol for the wild-type protein. The portal mutant exhibited a modest decrease (4-fold) in ligand binding affinity using the fluorescent probe 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS) as a surrogate ligand. 1,8-ANS displacement assays revealed that the binding affinity for oleate increased from a K0.5 of 196 +/- 15 nM for the wild-type protein to 165 +/- 8 for the portal mutant, while that for arachidonate decreased from the wild type of 186 +/- 11 nM to 418 +/- 26 nM for the portal mutant. To evaluate cavity accessibility, rate of 1,8-ANS binding was assessed between the portal and wild-type protein. Using equimolar amounts of ligand and protein at 4 degrees, 1,8-ANS bound within the cavity to 95% saturation (t0.95) in 750 ms, while the mutant protein was fully modified in less than 1.4 ms. To independently evaluate cavity accessibility, modification of the sole protein cysteine residue, C117 residing within the cavity near C2-C4 of the bound ligand, was monitored using 5,5'-dithio-bis(2-nitrobenzoic acid) modification. The half time for modification (t0.5) for the wild-type protein was approximately 20 s, while that for V32G F57G K58G occurred in less than a second. As such, enlargement of the portal region of FABP4 markedly increased the accessibility of ligands to the cavity while having only modest effects on ligand affinity. Taken together, these data provide support for the portal region hypothesis and suggest dynamic fluctuations in this region regulate cavity access, but not ligand affinity or selectivity.

Adipocytes↗
PubMed2004

Branch site haplotypes that control alternative splicing.

We show that the allele-dependent expression of transcripts encoding soluble HLA-DQbeta chains is determined by branchpoint sequence (BPS) haplotypes in DQB1 intron 3. BPS RNAs associated with low inclusion of the transmembrane exon in mature transcripts showed impaired binding to splicing factor 1 (SF1), indicating that alternative splicing of DQB1 is controlled by differential BPS recognition early during spliceosome assembly. We also demonstrate that naturally occurring human BPS point mutations that alter splicing and lead to recognizable phenotypes cluster in BP and in position -2 relative to BP, implicating impaired SF1-BPS interactions in disease-associated BPS substitutions. Coding DNA variants produced smaller fluctuations of exon inclusion levels than random exonic substitutions, consistent with a selection against coding mutations that alter their own exonization. Finally, proximal splicing in this multi-allelic reporter system was promoted by at least seven SR proteins and repressed by hnRNPs F, H and I, supporting an extensive antagonism of factors balancing the splice site selection. These results provide the molecular basis for the haplotype-specific expression of soluble DQbeta, improve prediction of intronic point mutations and indicate how extraordinary, selection-driven DNA variability in HLA affects pre-mRNA splicing.

Alternative Splicing↗
PubMed2006

Short- and long-term population dynamical consequences of asymmetric climate change in black grouse.

Temporal asymmetry in patterns of regional climate change may jeopardize the match between the proximate and ultimate cues of the timing of breeding. The consequences on short- and long-term population dynamics and trends as well as the underlying mechanisms are, however, often unknown. Using long-term data from Finland, we demonstrate that black grouse (Tetrao tetrix) have responded to spring warming by advancing both egg-laying and hatching. However, early summer (the time of hatching) has not advanced, and chicks have to face colder post-hatching conditions. Demonstrating that these conditions are critical to post-hatching survival, we show that chicks are increasingly suffering higher mortality because they hatch too early. Consequently, breeding success and population size has severely declined over the past four decades. Finally, we modelled the impact of this particular climate change scenario on population dynamics and show that the mismatch can further explain the observed collapse of cyclic fluctuations. Because the evolutionary response of grouse is lagging behind the novel selective pressures, seasonally asymmetric climate change is likely to constitute an important determinant of future short- and long-term changes in the dynamics of black grouse populations.

Adaptation, Physiological↗
PubMed2000

The coevolution of parasites with host-acquired immunity and the evolution of sex.

Here I present a deterministic model of the coevolution of parasites with the acquired immunity of their hosts, a system in which coevolutionary oscillations can be maintained. These dynamics can confer an advantage to sexual reproduction within the parasite population, but the effect is not strong enough to outweigh the twofold cost of sex. The advantage arises primarily because sexual reproduction impedes the response to fluctuating epistasis and not because it facilitates the response to directional selection-in fact, sexual reproduction often slows the response to directional selection. Where the cost of sexual reproduction is small, a polymorphism can be maintained between the sexuals and the asexuals. A polymorphism is maintained in which the advantage gained due to recombination is balanced by the cost of sex. At much higher costs of sex, a polymorphism between the asexual and sexual populations can still be maintained if the asexuals do not have a full complement of genotypes available to them, because the asexuals only outcompete those sexuals with which they share the same selected alleles. However, over time we might expect the asexuals to amass the full array of genotypes, thus permanently eliminating sexuals from the population. The sexuals may avoid this fate if the parasite population is finite. Although the model presented here describes the coevolution of parasites with the acquired immune responses of their hosts, it can be compared with other host-parasite models that have more traditionally been used to investigate Red Queen theories of the evolution of sex.

Animals↗
PubMed1995

Regulation of the alginate biosynthesis gene algC in Pseudomonas aeruginosa during biofilm development in continuous culture.

Reporter gene technology was used to observe the regulation of the alginate biosynthesis gene, algC in a mucoid strain of Pseudomonas aeruginosa in developing and mature biofilms in continuous culture on Teflon and glass substrata. The plasmid pNZ63, carrying an algC-lacZ transcriptional fusion, was shown to not be diluted in continuous culture over a period of 25 days in the absence of selection pressure. Biofilm cells under bulk phase steady-state conditions demonstrated fluctuations in algC expression over a 16-day period, but no trend of increased or decreased expression over the time interval was indicated. In vivo detection of algC up-expression in developing biofilms was performed with a fluorogenic substrate for the plasmid-borne lacZ gene product (beta-galactosidase) by using microscopy coupled with image analysis. By this technique, cells were tracked over time and analyzed for algC activity. During the initial stages of biofilm development, cells already attached to a glass surface for at least 15 min exhibited up-expression of algC, detectable as the development of whole-cell fluorescence. However, initial cell attachment to the substratum appeared to be independent of algC promoter activity. Furthermore, cells not exhibiting algC up-expression were shown to be less capable of remaining at a glass surface under flowing conditions than were cells in which algC up-expression was detected.

Alginates↗