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Persistent infection of mouse fibroblasts (L cells) with Chlamydia psittaci: evidence for a cryptic chlamydial form.

When monolayers of mouse fibroblasts (L cells) were infected with enough Chlamydia psittaci (strain 6BC) to destroy most of the host cells, 1 in every 10(5) to 10(6) originally infected cells gave rise to a colony of L cells persistently infected with strain 6BC. In these populations, the density of L cells and 6BC fluctuated periodically and reciprocally as periods of host cell increase were followed by periods of parasite multiplication. Successive cycles of L-cell and 6BC reproduction were sustained indefinitely by periodic transfer to fresh medium. Isolation of L cells and 6BC from persistent infections provided no evidence that there had been any selection of variants better suited for coexistence. Persistently infected populations consisting mainly of inclusion-free L cells yielded only persistently infected clones, grew more slowly, and cloned less efficiently. They were also almost completely resistant to superinfection with high multiplicities of either 6BC or the lymphogranuloma venereum strain 440L of Chlamydia trachomatis. These properties of persistently infected L cells may be accounted for by assuming that all of the individuals in these populations are cryptically infected with 6BC and that cryptic infection slows the growth of the host cell and makes it immune to infection with exogenous chlamydiae. According to this hypothesis, the fluctuations in host and parasite density occur because some factor periodically sets off the conversion of cryptic chlamydial forms into reticulate bodies that multiply and differentiate into infectious elementary bodies in a conventional chlamydial developmental cycle.

Animals↗

Voltage-gated cation conductance channel from fragmented sarcoplasmic reticulum: steady-state electrical properties.

The interaction of fragmented sarcoplasmic reticulum (SR) with an artificial planar phospholipid membrane under conditions known to induce fusion of phospholipid vesicles raises the conductance of the planar bilayer by several orders of magnitude. Measurements of steady-state electrical properties of bilayers thus modified by SR show that two types of conductance pathways are present. One is a voltage-independent pathway which may be somewhat anion-selective. The other is a voltage-gated ionophore showing selectivity to small monovalent cations. This latter ionophore is fully oriented within the artificial bilayer and is inhibited asymmetrically by divalent cations. It is also inhibited below pH6. The ionophore displays single-channel conductance fluctuations between two states. "open" and "closed", with an open-state conductance of 1.4 x 10-10 mho in 0.1 M K+. The physiological function of this ionophore is unknown.

Calcium↗

Ropinirole as an adjunct to levodopa in the treatment of Parkinson's disease: a 16-week bromocriptine controlled study.

BACKGROUND: and objectives Ropinirole is a non-ergoline, selective dopamine D(2) agonist. The aim of this study was to evaluate the efficacy and safety of ropinirole as an adjunct to levodopa in the treatment of Parkinson's disease (PD) complicated by motor fluctuations. METHODS: A total of 76 patients with PD (Hoehn and Yahr stage II to IV) were included in this trial. Each patient was randomly allocated to receive either ropinirole (n = 37) or bromocriptine (n = 39) as an adjunct to levodopa over a 16-week period. Ropinirole and bromocriptine were titrated for optimal efficacy and tolerability. This optimal dose was then maintained for the rest of the study. Response rate was defined as the percentage of patients who achieved at least a 20 % reduction in levodopa dose. Clinical status was also assessed using the Unified Parkinson's Disease Rating Scale (UPDRS), Clinical Global Impression (CGI), and reduction in time spent 'off'. RESULTS: Ropinirole produced a significantly greater response rate than bromocriptine (odds ratio 2.995, 95 % C. I. (1.157, 7.751) p < 0.05). There was also a statistically significant difference between the groups in the proportion of patients who were 'improved' on the CGI improvement scale (91.9 % for ropinirole, 74.3 % for bromocriptine, p = 0.046). Other measures, including at least a 20 % improvement in the UPDRS motor score (70 % for ropinirole and 63.3 % for bromocriptine), and a 20 % reduction in 'off' duration (81 % for ropinirole and 52.4 % for bromocriptine) showed a trend in favour of ropinirole. There was no significant difference between the two groups in the overall incidence of adverse effects (ropinirole, 59.5 %; bromocriptine, 59 %). In each group, the most common side-effects were dizziness, dyskinesia and nausea/vomiting. No patients were withdrawn from the study because of side-effects. CONCLUSION: Ropinirole was found to be safe and well-tolerated. Ropinirole as an adjunct to levodopa in the treatment of PD with motor fluctuation was associated with more significant reduction of levodopa dose and, on one form of analysis, with significantly greater improvement in CGI ratings than bromocriptine. On the other efficacy measures the two drugs were comparable.

Antiparkinson Agents↗

An open-label, multicentre clinical trial to determine the levodopa dose-sparing capacity of pramipexole in patients with idiopathic Parkinson's disease.

Ninety-three patients with idiopathic Parkinson's disease (PD) entered a 12 week open-label, baseline controlled, multicentre study. The study was designed to determine the levodopa sparing effect of pramipexole as add-on treatment in PD while maintaining an optimal clinical improvement in motor performance. The overall reduction in adjusted levodopa dose was the primary endpoint. Unified Parkinson's Disease Rating Scale (UPDRS) subscores as well as motor fluctuations, frequency and severity of dyskinesia (assessed by patient diaries) were secondary endpoints. Pramipexole permitted a median reduction of adjusted levodopa by 40% while maintaining or improving the UPDRS scores in 61 patients (per protocol [PP] analysis). The intent-to-treat (ITT) analysis (90 patients) similarly revealed a median reduction of 40%. An anticipated short-term levodopa dose reduction as substantiated by 95% confidence interval calculations lies within a range of 35% to 50%. If unadjusted levodopa doses were considered, a median reduction of 42% (PP) or 43% (ITT) was achieved. 47% patients (ITT) had a levodopa dose reduction (adjusted) of more than 40% while maintaining or improving their level of efficacy, and 72.2% had a reduction of at least 20%. Motor fluctuations improved compared to baseline according to patient diaries and UPDRS part IV. These findings suggest that pramipexole can markedly reduce the daily levodopa dosage without deterioration of motor response and support that this new selective D2/D3 receptor agonist also improves later levodopa-associated motor complications.

Aged↗

High-frequency components in epileptiform EEG.

We analyzed the spectral content of interictal and ictal EEG of rats treated with metrazol. Significant power was present in the EEG between 100 and 800 Hz ictally but not interictally. Since high-frequency activities appeared to accompany spiking episodes, a computer algorithm was developed to recognize rapid voltage fluctuations or high-frequency components in EEG. The algorithm turned out to be effective in the identification of seizure spikes. The significance of high frequencies in EEG spikes and the advantage of the spike selection algorithm are discussed.

Algorithms↗

The critical period for wing pattern induction in the polyphenic tropical butterfly Bicyclus anynana (Satyrinae).

Adults of the butterfly Bicyclus anynana express striking phenotypic plasticity. A wet season form has conspicuous marginal eyespots and a medial pale band which are much reduced in the dry season form. These alternative forms are produced after rearing at high or low temperatures, respectively. We used 'window' experiments involving switching of larvae and pupae between high and low temperatures at different stages during development to examine the timing of sensitivity to environmental temperature. The final, fifth larval instar is shown to be especially sensitive. The fourth larval instar and the very early pupal period are also sensitive. It is argued that an increasing sensitivity during growth is ecologically adaptive since the late larval environment will be the most accurate predictor for the adult environment in which the wing phenotype is subject to selection. The period of sensitivity is not as short as a few days. This may minimize the chance of any 'mistakes' in matching the adult phenotype to the season because of short-term environmental fluctuations during the larval period. The observed sensitivity occurs as late as possible during growth since the wing pattern is developmentally determined at the end of the early part of the pupal stage.

Journal Article↗

Self-organization of a propulsive actin network as an evolutionary process.

The leading edge of motile cells is propelled by polymerization of actin filaments according to a dendritic nucleation/array treadmilling mechanism. However, little attention has been given to the origin and maintenance of the dendritic array. Here we develop and test a population-kinetics model that explains the organization of actin filaments in terms of the reproduction of dendritic units. The life cycle of an actin filament consists of dendritic nucleation on another filament (birth), elongation by addition of actin subunits and, finally, termination of filament growth by capping protein (death). The regularity of branch angle between daughter and mother filaments endows filaments with heredity of their orientation. Fluctuations of branch angle that become fixed in the actin network create errors of orientation (mutations) that may be inherited. In our model, birth and death rates depend on filament orientation, which then becomes a selectable trait. Differential reproduction and elimination of filaments, or natural selection, leads to the evolution of a filament pattern with a characteristic distribution of filament orientations. We develop a procedure based on the Radon transform for quantitatively analyzing actin networks in situ and show that the experimental results are in agreement with the distribution of filament orientations predicted by our model. We conclude that the propulsive actin network can be understood as a self-organizing supramolecular ensemble shaped by the evolution of dendritic lineages through natural selection of their orientation.

Actins↗

Is there a dynamic DNA-protein interface in the transcription factor IIIA-5 S rRNA gene complex?

Others have proposed that several amino acid side chains exhibit considerable conformational mobility at the DNA-protein interface in the transcription factor IIIA.5 S rRNA gene complex and that the rapid movements of these side chains permit them to make fluctuating contacts with adjacent bp in the DNA target site. This "dynamic interface" model makes biochemical predictions concerning the consequences of truncating specific amino acid side chains and the effects of these truncations on sequence selectivity in DNA binding. The model also makes predictions concerning the effects of DNA sequence context on the apparent energetic contributions to binding made by individual bp. We have tested these predictions, and our results are inconsistent with any significant energetic role being played by the contact of multiple bp by conformationally mobile amino acid side chains. They do, however, show that some individual amino acids affect the recognition of multiple bp through mechanisms other than direct interaction.

Animals↗

A reconstruction method of surface morphology with genetic algorithms in the scanning electron microscope.

A reconstruction method of surface morphology with genetic algorithms from a line scan in the scanning electron microscope (SEM) has been developed. This method is an application of genetic algorithms, which are search algorithms based on the mechanics of natural selection and genetics, to surface morphology in SEM. To reconstruct the microstructures of a specimen surface on a substrate, the system requires only a line scan taken by a conventional SEM. To deal with statistical fluctuations in the grey level of each pixel on the experimental line scan, the reduced chi-square distribution is used as the objective function and a scheme for minimizing the number of vertices in the reconstructed surface profile is adopted. The resolution in the Z-direction tends to be statistical and independent of the angle of the specimen surface.

Journal Article↗

Beta1-adrenergic blockade augments pulsatile PTH secretion in humans.

Pulsatile peptide hormone secretion provides efficient control of specific end organ functions. To test the hypothesis that sympathetic neuronal activity drives synchronous pulsatile PTH release from the parathyroid glands, we investigated the acute effects of beta1-adrenergic receptor blockade on PTH secretion patterns in a single-blinded study in nine healthy adults. Plasma PTH levels were determined at 1-min intervals. After a 75-min baseline period, seven subjects received a continuous intravenous infusion of the short-acting beta1-adrenergic receptor blocker esmolol for 105 min. After a 30-min washout period, esmolol was infused for another 30 min. Two additional subjects were randomized to receive solvent infusions. PTH secretion characteristics were analyzed by multiparameter deconvolution analysis, and the orderliness of plasma PTH fluctuations by Approximate Entropy statistics. BP slightly decreased during esmolol infusion, whereas heart rate, ionized calcium, phosphate, magnesium, and plasma and urine catecholamines remained unchanged. Esmolol increased mean plasma PTH by 33 +/- 8% (P < 0.01), due to a preferential increase in the pulsatile PTH secretion component (+129 +/- 44%, P < 0.02). The increased pulsatile PTH secretion was mediated by an augmented PTH burst mass (+117 +/- 42%, P < 0.01), whereas burst frequency remained unchanged. The regularity of PTH fluctuations was not affected by the beta-adrenergic blockade. The effects were reproducible during the second esmolol infusion. The authors conclude that the sympathetic nervous system has a modulating effect on pulsatile PTH secretion. Selective beta-1 adrenergic blockade acutely increases plasma PTH by augmenting the mass of hormone secreted per burst, but it does not alter the rhythmicity of pulsatile PTH release.

Adrenergic beta-Antagonists↗

Density distribution of hematopoietic progenitor cells and T lymphocytes from peripheral blood: heterogeneity of the human population.

Density gradient centrifugation has been used in animal systems to purify stem cells and eliminate T lymphocytes prior to allogeneic transplantation. There is substantial disagreement whether the same approach can be used to purify hematopoietic stem cells obtained from human peripheral blood. The purpose of the present study was to resolve that issue by determining the density distribution of 4 classes of human leucocytes: total mononuclear cells, T lymphocytes, CFUc, and BFUe. To ensure a representative sampling, a large number of randomly selected donors were analyzed. The results show that most T lymphocytes band between 1.068 and 1.071 g/ml, with relatively little variation from individual to individual. In contrast, the density distributions of both CFUc and BFUe fluctuated markedly from donor to donor. As a consequence, there was significant variability in the degree of progenitor-T cell separation. The implications of these results for clinical application of the density separation technique are discussed.

Cell Separation↗

Ancient Introgression Explains Mitochondrial Genome Capture and Mitonuclear Discordance Among South American Collared Tropidurus Lizards.

Mitonuclear discordance-evolutionary discrepancies between mitochondrial and nuclear DNA phylogenies-can arise from various factors, including introgression, incomplete lineage sorting, recent or ancient demographic fluctuations, sex-biased dispersal asymmetries, among others. Understanding this phenomenon is crucial for accurately reconstructing evolutionary histories, as failing to account for discordance can lead to misinterpretations of species boundaries, phylogenetic relationships, and historical biogeographic patterns. We investigate the evolutionary drivers of mitonuclear discordance in the Tropidurus spinulosus species group, which contains nine species of lizards inhabiting open tropical and subtropical environments in South America. Using a combination of population genetic and phylogenomic approaches applied to mitochondrial and nuclear data, we identified different instances of gene flow that occurred in ancestral lineages of extant species. Our results point to a complex evolutionary history marked by prolonged isolation between species, demographic fluctuations, and potential episodes of secondary contact with genetic admixture. These conditions likely facilitated mitochondrial genome capture while diluting signals of nuclear introgression. Furthermore, we found no strong evidence supporting incomplete lineage sorting or natural selection as primary drivers of the observed mitonuclear discordance. Therefore, the unveiled patterns are most consistent with neutral demographic processes, coupled with ancient mitochondrial introgression, as the main factors underlying the mismatch between nuclear and mitochondrial phylogenies in this system. Future research could further explore the role of other demographic processes, such as asymmetric sex-biased dispersal, in shaping these complex evolutionary patterns.

Animals↗

Phenotypic hiding: the carryover of mutations in RNA viruses as shown by detection of mar mutants in influenza virus.

When influenza virus monoclonal antibody-resistant (mar) mutants are selected by incubation in vitro with excess antibody, 90 to 99% of the mutants are not detectable. This observation may be explained by encapsidation of mar mutant RNAs within phenotypically wild-type envelopes. This phenotypic hiding can be revealed by selection of mar mutants in vivo after virus uncoating. Using experimental procedures appropriate to detect all viable mar mutants in a virus population, we determined precisely the mutation rates to the mar genotype by the fluctuation test for two nonoverlapping monoclonal antibodies.

Antibodies, Monoclonal↗

[Mistletoe therapy from the pharmacologic perspective].

Because of their cytostatic/apoptotic and immunomodulatory effects mistletoe extracts are often applied in tumour patients. Recent experimental data suggest that the mistletoe lectins Viscum album agglutinin (VAA)-I and -II are play an important role in the efficacy of mistletoe therapy. VAA-I and -II are members of the type-II ribosome-inactivating proteins. VAA-I has been shown to induce cytostatic effects in cultures of various eukaryotic cells in vitro. In 24-hour cultures of human peripheral lymphocytes, flow-cytometric investigations with propidium iodide (PI) in hypotonic buffer and quantitative assessment of DNA breaks with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end-labeling (TUNEL) assay were carried out; they revealed a dose-dependent VAA-I-induced apoptosis (lectin concentrations between 10 ng/ml and 1 microg/ml). In 24-hour cultures of peripheral blood mononuclear cells (PBMC), VAA-I in non-cytotoxic concentrations (1+/-10 ng/ml) induced mRNA expression and enhanced the secretion of proinflammatory cytokines. The stimulation of NK cells by VAA-I in vitro was enhanced in additive manner by the combination of VAA-I with IL-2 and IL-12. In culture of PBMC and bone marrow CD34+ cells coincubation of VAA-I with other haematopoietic growth factors induced a dose-dependent increase in clonogenic growth. In cancer patients the mechanisms of natural immunity, believed to be essential for their survival, are often significantly decreased. VAA-I and standardized mistletoe extracts are able to stimulate the cellular parameters of natural immunity with a bell-shaped curve of efficacy. Studies in animal models suggest that application of 0.5-3 ng/kg VAA-I twice a week is effective in sustaining the elevation of the number and activity of peripheral blood NK cells. These parameters often exhibit high intrinsic fluctuations, in healthy persons, however, blind crossover studies reveal an optimal lectin dose of about 0.5 and 1 ng/kg bw, suggesting a potential use of mistletoe preparations as a modulator of the natural immune system. Selective apoptotic effects of VAA-I may represent a novel approach for pharmacological manipulation of the balance between cell growth and programmed cell death. Appropriate combination of immunomodulatory and cytotoxic doses may open new clinical perspectives in the mistletoe therapy.

Bone Marrow Cells↗

Sleep and exertional periodic breathing in chronic heart failure: prognostic importance and interdependence.

BACKGROUND: Sleep and exertional periodic breathing are proverbial in chronic heart failure (CHF), and each alone indicates poor prognosis. Whether these conditions are associated and whether excess risk may be attributed to respiratory disorders in general, rather than specifically during sleep or exercise, is unknown. METHODS AND RESULTS: We studied 133 CHF patients with left ventricular ejection fraction (LVEF) < or =40%. During 1170+/-631 days of follow-up, 31 patients (23%) died. Nonsurvivors had higher New York Heart Association class, ventilatory response (ve/vco2 slope), and apnea-hypopnea index (AHI) and lower peak vo2 (all P<0.01); lower LVEF and prescription of beta-blockers, and shorter transmitral deceleration time (all P<0.05). Exertional oscillatory ventilation (EOV), established by cyclic fluctuations in minute ventilation that persisted for > or =60% of exercise duration with an amplitude > or =15% of the average resting value, was significantly more frequent in nonsurvivors (42% versus 15%, P<0.01). Multivariable analysis selected AHI (hazard ratio [HR] 5.66, 95% CI 2.3 to 19.9, P<0.01), peak vo2 (HR 0.93, 95% CI 0.90 to 0.97, P<0.01), and beta-blocker prescription (HR 0.34, 95% CI 0.13 to 0.87, P<0.05) as predictors of cardiac events. The best cutoff for AHI was >30/h. EOV was significantly related to AHI >30/h (chi2 14.6, P<0.01): 78% of EOV patients showed AHI >30/h. Multivariable analysis, including breathing disorders alone (EOV, AHI >30/h) or in combination (EOV plus AHI >30/h), selected combined disorders as the strongest predictor of events (HR 6.65, 95% CI 2.6 to 17.1, P<0.01). CONCLUSIONS: In CHF, EOV is significantly associated with AHI >30/h. Although each breathing disorder alone is linked to total mortality, their combination has a crucial prognostic burden.

Aged↗

[Effect of different factors on the sex ratio in laboratory mouse embryogenesis].

Influence of two factors on distribution of embryos according to the sex is considered: genetic peculiarities of inbred mice and hybrid mice, as well as spontaneous embryonal death of the embryos of different sex. The work has been performed on C3H, CBA, C57BL strains of mice and on hybrid mice of CBA/C57BL and ICR/In(X)1H strains. The sex ratio in the mice studied is balanced. In the CBA/C57BL hybrid mice a certain shift in the sex ratio is noted towards female embryos. In the offspring of the ICR/In(X)1H hybrid mice the sex distribution of the embryos does not differ from the ratio 1:1. The results of the investigation confirm the opinion of the literature that genetic peculiarities of the inbred mice do not essentially influence the sex ratio in their offspring, while fluctuation in the sex ratio in the hybrid mice are evidently connected, in a greater degree, with their genetic peculiarities. As demonstrates the analysis of the sex relation and the embryonal death, there is no selective death in the embryos of any sex during embryogenesis in the mice investigated.

Animals↗

Computer model of the maintenance and selection of genetic heterogeneity in polygamous helminths.

A stochastic simulation model of the transmission and maintenance of genetic heterogeneity in the absence and presence of external selection pressures is presented for polygamous intestinal helminths such as Ascaris. The model assumes that the density distribution of the adult parasites is highly aggregated and that density-dependent effects on fecundity are important. The model gives rise to stable infection rates in the host. Where the parasite population contains genetic heterogeneity, with the exception of stochastic fluctuations which models genetic drift, the ratio of the different alleles remained constant over extended periods of time. This result contrasts with that of an earlier analytical model (Anderson, R.M., May, M.R. & Gutpa S. (1989) Parasitology 99, S59-S79), in which uneven mating probabilities for the different combinations of worm possible in a host was postulated to inevitably lead to fixation of the most abundant allele. New results suggest that in spite of the restricted choice of mating available to a worm in the confines of a host, selection pressure always leads to enrichment of the parasites carrying resistant alleles.

Animals↗

Subcellular distribution, molecular dynamics and catabolism of plasmalogens in myocardium.

Recent studies have implicated accelerated sarcolemmal phospholipid catabolism as a mediator of the lethal sequelae of atherosclerotic heart disease. We have demonstrated that plasmalogens are the predominant phospholipid constituents of canine myocardium and that plasmalogens are hydrolyzed by a novel calcium independent plasmalogen selective phospholipase A2. Since the activities of phospholipases are modulated by the molecular dynamics and interfacial characteristics of their phospholipid substrates, we compared the molecular dynamics of plasmenylcholine and phosphatidylcholine vesicles by electron spin resonance spectroscopy and deuterium magnetic resonance spectroscopy. Plasmenylcholine vesicles have separate and distinct molecular dynamics in comparisons to their phosphatidylcholine counterparts as ascertained by substantial decreases in the angular fluctuations and motional velocities of probes attached to their sn-2 aliphatic constituents. Furthermore, since free radical oxidation of myocardial lipid constituents occurs during myocardial ischemia and reperfusion, we demonstrated that 1O2 mediated oxidation of plasmenylcholine resulted in the generation of several products which have chromatographic characteristics and molecular masses corresponding to 2-acyl lysophosphatide derivatives. Taken together, these studies underscore the biologic significance of the predominance of sarcolemmal plasmalogens present in mammalian myocardium and suggest that their catabolism by plasmalogen selective phospholipases and/or oxidative processes may contribute to the lethal sequelae of myocardial ischemia.

Animals↗