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Sample records for cryptomonas paramecium reduction

  1. The Complete Plastid Genome Sequence of the Secondarily Nonphotosynthetic Alga Cryptomonas paramecium: Reduction, Compaction, and Accelerated Evolutionary Rate

    PubMed Central

    Donaher, Natalie; Tanifuji, Goro; Onodera, Naoko T.; Malfatti, Stephanie A.; Chain, Patrick S. G.; Hara, Yoshiaki

    2009-01-01

    The cryptomonads are a group of unicellular algae that acquired photosynthesis through the engulfment of a red algal cell, a process called secondary endosymbiosis. Here, we present the complete plastid genome sequence of the secondarily nonphotosynthetic species Cryptomonas paramecium CCAP977/2a. The ∼78 kilobase pair (Kbp) C. paramecium genome contains 82 predicted protein genes, 29 transfer RNA genes, and a single pseudogene (atpF). The C. paramecium plastid genome is approximately 50 Kbp smaller than those of the photosynthetic cryptomonads Guillardia theta and Rhodomonas salina; 71 genes present in the G. theta and/or R. salina plastid genomes are missing in C. paramecium. The pet, psa, and psb photosynthetic gene families are almost entirely absent. Interestingly, the ribosomal RNA operon, present as inverted repeats in most plastid genomes (including G. theta and R. salina), exists as a single copy in C. paramecium. The G + C content (38%) is higher in C. paramecium than in other cryptomonad plastid genomes, and C. paramecium plastid genes are characterized by significantly different codon usage patterns and increased evolutionary rates. The content and structure of the C. paramecium plastid genome provides insight into the changes associated with recent loss of photosynthesis in a predominantly photosynthetic group of algae and reveals features shared with the plastid genomes of other secondarily nonphotosynthetic eukaryotes. PMID:20333213

  2. Paramecium

    SciTech Connect

    Gortz, H.D.

    1988-01-01

    This book presents a survey of the current knowledge and research on Paramecium. Scientists find Paramecium a most useful object for the study of basic biological enigmas. To cover all aspects on the cell biology, cell physiology, genetics, developmental biology, ecology, endocytobiology, and molecular biology of Paramecium, specialists were asked to review their own fields. They have also included quite recent studies on the cytoskeleton, endo- and exocytosis, aging, sensory and membrane physiology, electrophysiology and motility.

  3. Nuclear transplant studies on the reduction in numbers of presumptive germ nuclei in exconjugants of Paramecium caudatum.

    PubMed

    Mikami, K

    1982-08-01

    Nuclear differentiation in exconjugants of Paramecium caudatum is closely associated with a brief localization of the postzygotic nuclei near the opposite ends of the cell, with the germinal nucleus (micronucleus) in the anterior region and the somatic nuclei (macronuclei) in the posterior region. The posterior nuclei cannot regenerate to produce micronuclei when all four anterior nuclei are removed. There is no difference among the anterior four presumptive micronuclei, because, when any three of them were removed, the remaining nucleus was able to divide at each postconjugational fission and to persist as a micronucleus during the vegetative phase. This conclusion agrees with the results of transplanting a presumptive micronucleus into a vegetative cell. Cells during the vegetative phase, however, normally have only one micronucleus. Micronuclear number must be reduced to arrive at the uni-micronucleate condition after the stage of macro- and micronuclear differentiation. Elimination of supernumerary presumptive micronuclei, which had been indicated by morphological observations, was confirmed by the results of nuclear transplantation studies.

  4. Homology-dependent Gene Silencing in Paramecium

    PubMed Central

    Ruiz, Françoise; Vayssié, Laurence; Klotz, Catherine; Sperling, Linda; Madeddu, Luisa

    1998-01-01

    Microinjection at high copy number of plasmids containing only the coding region of a gene into the Paramecium somatic macronucleus led to a marked reduction in the expression of the corresponding endogenous gene(s). The silencing effect, which is stably maintained throughout vegetative growth, has been observed for all Paramecium genes examined so far: a single-copy gene (ND7), as well as members of multigene families (centrin genes and trichocyst matrix protein genes) in which all closely related paralogous genes appeared to be affected. This phenomenon may be related to posttranscriptional gene silencing in transgenic plants and quelling in Neurospora and allows the efficient creation of specific mutant phenotypes thus providing a potentially powerful tool to study gene function in Paramecium. For the two multigene families that encode proteins that coassemble to build up complex subcellular structures the analysis presented herein provides the first experimental evidence that the members of these gene families are not functionally redundant. PMID:9529389

  5. Magnetic field influence on paramecium motility

    SciTech Connect

    Rosen, M.F.; Rosen, A.D. )

    1990-01-01

    The influence of a moderately intense static magnetic field on movement patterns of free swimming Paramecium was studied. When exposed to fields of 0.126 T, these ciliated protozoa exhibited significant reduction in velocity as well as a disorganization of movement pattern. It is suggested that these findings may be explained on the basis of alteration in function of ion specific channels within the cell membrane.

  6. Endosymbionts in paramecium.

    PubMed

    Fujishima, Masahiro; Kodama, Yuuki

    2012-05-01

    Paramecium species are extremely valuable organisms to enable experiments for the reestablishment of endosymbiosis. This is investigated in two different systems, the first with Paramecium caudatum and the endonuclear symbiotic bacterium Holospora species. Although most endosymbiotic bacteria cannot grow outside the host cell as a result of their reduced genome size, Holospora species can maintain their infectivity for a limited time. We found that an 89-kDa periplasmic protein has an important function for Holospora's invasion into the target nucleus, and that Holospora alters the host gene expression; the host thereby acquires resistance against various stresses. The second system is the symbiosis between P. bursaria and symbiotic Chlorella. Alga-free P. bursaria and the algae retain the ability to grow without a partner. Consequently, endosymbiosis between the aposymbiotic host cells and the symbiotic algae can be reestablished easily by mixing them. We now found four checkpoints for the reestablishment of the endosymbiosis between P. bursaria and the algae. The findings in the two systems provide excellent opportunities for us to elucidate not only infection processes but also to assess the associations leading to eukaryotic cell evolution. This paper summarizes recent progresses on reestablishment of the primary and the secondary endosymbiosis in Paramecium. Copyright © 2011 Elsevier GmbH. All rights reserved.

  7. Is paramecium swimming autonomic?

    NASA Astrophysics Data System (ADS)

    Bandyopadhyay, Promode R.; Toplosky, Norman; Hansen, Joshua

    2010-11-01

    We seek to explore if the swimming of paramecium has an underlying autonomic mechanism. Such robotic elements may be useful in capturing the disturbance field in an environment in real time. Experimental evidence is emerging that motion control neurons of other animals may be present in paramecium as well. The limit cycle determined using analog simulation of the coupled nonlinear oscillators of olivo-cerebellar dynamics (ieee joe 33, 563-578, 2008) agrees with the tracks of the cilium of a biological paramecium. A 4-motor apparatus has been built that reproduces the kinematics of the cilium motion. The motion of the biological cilium has been analyzed and compared with the results of the finite element modeling of forces on a cilium. The modeling equates applied torque at the base of the cilium with drag, the cilium stiffness being phase dependent. A low friction pendulum apparatus with a multiplicity of electromagnetic actuators is being built for verifying the maps of the attractor basin computed using the olivo-cerebellar dynamics for different initial conditions. Sponsored by ONR 33.

  8. Symbiosis in Paramecium Bursaria.

    PubMed

    Karakashian, M W

    1975-01-01

    Paramecium bursaria normally appears green dut to several hundred symbiotic Chlorella which are dispersed throughout its cytoplasm. The symbionts are situated within individual vacuoles and these alga-vacuole complexes grow and divide at a rate compatible with that of the paramecium. The symbiotic units also persist through conjugation and the subsequent reorganization of the host. Studies of the benefit of the symbiosis to the ciliate hosts have shown that they are able to grow and survive better than aposymbiotic animals in environments deficient in bacteria. The symbionts are also able to extract nourishment from the host when it is well fed and they are deprived of light. The biochemical nature of these exchanges has not been determined. Potential symbionts usually enter the host in food vacuoles. If they are ingested in sufficient numbers, they are able to interfere with the normal course of host digestion, perhaps by preventing the release of digestive enzymes into the food vacuole. All natural symbionts of P. bursaria appear able to reinfect aposymbiotic cells. Some freeliving strains of Chlorella and related algae are also infective, but these associations are relatively unstable and provide little evident benefit to the host. Host susceptibility to infection by certain strains of free-living algae is invariably lost with time. This loss is specific and often rapid, but it does not occur simultaneously in subcultures derived from the original susceptible culture. The basis for these susceptibility changes is still unknown, but they may be related to long-lasting effect of the previous symbionts on the digestive efficiency of the paramecium host.

  9. Effects of light intensity on the growth of Cryptomonas sp. (Cryptophyceae)

    NASA Astrophysics Data System (ADS)

    Weng, Huan-Xin; Qin, Ya-Chao; Sun, Xiang-Wei; Chen, Xun-Hong; Chen, Jing-Feng

    2009-03-01

    Laboratory culture experiments have been conducted to evaluate the effects of light intensity on the growth of Cryptomonas sp. (Cryptophyceae) and the discrepancy in absorption of iron and phosphorus under different light conditions. Results show that there is an exponential correlation between algal growth rate and light intensity. The saturating and semi-saturating light values for Cryptomonas sp. cells are 150 and 47 μmol photons m-2 s-1, respectively. More uptake of Fe, P, and other trace elements such as Zn, Mn, Co, and Mo is observed in the low light cultures, although the algal growth rates are slow. The growth rate at 10 μmol photons m-2 s-1 is only 10% of that at 150 μmol photons m-2 s-1, whereas Fe and P uptake increases by 150 and 100%, respectively. These results suggest potential implications of differentiation in absorption of iron and phosphorus at different light intensities for the occurrence of harmful algal blooms (HABs). The mechanisms of light intensity regulating nutrient uptake as well as the occurrence of HABs are also discussed.

  10. Localization of calcium channels in Paramecium caudatum.

    PubMed Central

    Dunlap, K

    1977-01-01

    1. Electrical recordings from Paramecium caudatum were made after removal of the cilia with chloral hydrate and during ciliary regrowth to study the electrical properties of that portion of the surface membrane enclosing the ciliary axoneme. 2. Removal of the somatic cilia (a 50% reduction in membrane surface area) results in an almost complete elimination of the regenerative Ca response, all-or-none Ba2+ spike, and delayed rectification. 3. A twofold increase in input resistance resulted from the 50% reduction in membrane surface area. 4. The electrical properties remained unchanged, despite prolonged exposure to the chloral hydrate, until the cilia were mechanically removed. 5. Restoration of the Ca response accompanied ciliary regrowth, so that complete excitability returns when the cilia regain their original lengths. 6. It is concluded that the voltage-sensitive Ca channels are localized to that portion of surface membrane surrounding the cilia. 7. Measurements of membrane constants before and after deciliation and estimations of the cable constants of a single cilium suggest that the cilia of Paramecium may be fully isopotential along their length and with the major cell compartment. Images Plate 1 Plate 2 PMID:915829

  11. Effects of light intensity and temperature on Cryptomonas ovata (Cryptophyceae) growth and nutrient uptake rates

    USGS Publications Warehouse

    Cloern, James E.

    1977-01-01

    Specific growth rate of Cryptomonas ovata var. palustris Pringsheim was measured in batch culture at 14 light-temperature combinations. Both the maximum growth rate (μm) and optimum light intensity (Iopt) fit an empirical function that increases exponentially with temperature up to an optimum (Topt), then declines rapidly as temperature exceeds Topt. Incorporation of these functions into Steele's growth equation gives a good estimate of specific growth rate over a wide range of temperature and light intensity. Rates of phosphate, ammonium and nitrate uptake were measured separately at 16 combinations of irradiance and temperature and following a spike addition of all starved cells initially took up nutrient at a rapid rate. This transitory surge was followed by a period of steady, substrate-saturated uptake that persisted until external nutrient concentration fell. Substrate-saturated NO3−-uptake proceeded at very slow rates in the dark and was stimulated by both increased temperature and irradiance; NH4+-uptake apparently proceeded at a basal rate at 8 and l4 C and was also stimulated by increased temperature and irradiance. Rates of NH4−-uptake were much higher than NO3−-uptake at all light-temperature combinations. Below 20 C, PO4−3-uptake was more rapid in dark than in light, but was light enhanced at 26 C.

  12. Population Genomics of Paramecium Species.

    PubMed

    Johri, Parul; Krenek, Sascha; Marinov, Georgi K; Doak, Thomas G; Berendonk, Thomas U; Lynch, Michael

    2017-05-01

    Population-genomic analyses are essential to understanding factors shaping genomic variation and lineage-specific sequence constraints. The dearth of such analyses for unicellular eukaryotes prompted us to assess genomic variation in Paramecium, one of the most well-studied ciliate genera. The Paramecium aurelia complex consists of ∼15 morphologically indistinguishable species that diverged subsequent to two rounds of whole-genome duplications (WGDs, as long as 320 MYA) and possess extremely streamlined genomes. We examine patterns of both nuclear and mitochondrial polymorphism, by sequencing whole genomes of 10-13 worldwide isolates of each of three species belonging to the P. aurelia complex: P. tetraurelia, P. biaurelia, P. sexaurelia, as well as two outgroup species that do not share the WGDs: P. caudatum and P. multimicronucleatum. An apparent absence of global geographic population structure suggests continuous or recent dispersal of Paramecium over long distances. Intergenic regions are highly constrained relative to coding sequences, especially in P. caudatum and P. multimicronucleatum that have shorter intergenic distances. Sequence diversity and divergence are reduced up to ∼100-150 bp both upstream and downstream of genes, suggesting strong constraints imposed by the presence of densely packed regulatory modules. In addition, comparison of sequence variation at non-synonymous and synonymous sites suggests similar recent selective pressures on paralogs within and orthologs across the deeply diverging species. This study presents the first genome-wide population-genomic analysis in ciliates and provides a valuable resource for future studies in evolutionary and functional genetics in Paramecium. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. L-glutamate Receptor In Paramecium

    NASA Astrophysics Data System (ADS)

    Bernal-Martínez, Juan; Ortega-Soto, Arturo

    2004-09-01

    Behavioral, electrophysiological and biochemical experiments were performed in order to establish the presence of a glutamate receptor in the ciliate Paramecium. It was found that an AMPA/KA receptor is functionally expressed in Paramecium and that this receptor is immunologically and fillogenetically related to the AMPA/KA receptor present in vertebrates.

  14. Swimming of Paramecium in confined channels

    NASA Astrophysics Data System (ADS)

    Jung, Sunghwan

    2012-02-01

    Many living organisms in nature have developed a few different swimming modes, presumably derived from hydrodynamic advantage. Paramecium is a ciliated protozoan covered by thousands of cilia with a few nanometers in diameter and tens of micro-meters in length and is able to exhibit both ballistic and meandering motions. First, we characterize ballistic swimming behaviors of ciliated microorganisms in glass capillaries of different diameters and explain the trajectories they trace out. We develop a theoretical model of an undulating sheet with a pressure gradient and discuss how it affects the swimming speed. Secondly, investigation into meandering swimmings within rectangular PDMS channels of dimension smaller than Paramecium length. We find that Paramecium executes a body-bend (an elastic buckling) using the cilia while it meanders. By considering an elastic beam model, we estimate and show the universal profile of forces it exerts on the walls. Finally, we discuss a few other locomotion of Paramecium in other extreme environments like gel.

  15. Paramecium swimming in capillary tube

    NASA Astrophysics Data System (ADS)

    Jana, Saikat; Um, Soong Ho; Jung, Sunghwan

    2012-04-01

    Swimming organisms in their natural habitat need to navigate through a wide range of geometries and chemical environments. Interaction with boundaries in such situations is ubiquitous and can significantly modify the swimming characteristics of the organism when compared to ideal laboratory conditions. We study the different patterns of ciliary locomotion in glass capillaries of varying diameter and characterize the effect of the solid boundaries on the velocities of the organism. Experimental observations show that Paramecium executes helical trajectories that slowly transition to straight lines as the diameter of the capillary tubes decreases. We predict the swimming velocity in capillaries by modeling the system as a confined cylinder propagating longitudinal metachronal waves that create a finite pressure gradient. Comparing with experiments, we find that such pressure gradient considerations are necessary for modeling finite sized ciliary organisms in restrictive geometries.

  16. Effect of Paramecium biaurelia cytoplasm transplantation on the duration of the interautogamous interval (IAI) in Paramecium tetraurelia.

    PubMed

    Prajer, M

    1994-01-01

    The cytoplasm of Paramecium biaurelia in various stages of IAI was transplanted to Paramecium tetraurelia cells of the same clonal age. Such donor cytoplasm was effective in retardation of the expression of autogamy in the recipient clones. The results suggest that the cytoplasm of Paramecium biaurelia may contain the autogamous immaturity factor whose level changes during the run of IAI and whose specificity is the same as in Paramecium tetraurelia.

  17. ParameciumDB: a community resource that integrates the Paramecium tetraurelia genome sequence with genetic data.

    PubMed

    Arnaiz, Olivier; Cain, Scott; Cohen, Jean; Sperling, Linda

    2007-01-01

    ParameciumDB (http://paramecium.cgm.cnrs-gif.fr) is a new model organism database associated with the genome sequencing project of the unicellular eukaryote Paramecium tetraurelia. Built with the core components of the Generic Model Organism Database (GMOD) project, ParameciumDB currently contains the genome sequence and annotations, linked to available genetic data including the Gif Paramecium stock collection. It is thus possible to navigate between sequences and stocks via the genes and alleles. Phenotypes, of mutant strains and of knockdowns obtained by RNA interference, are captured using controlled vocabularies according to the Entity-Attribute-Value model. ParameciumDB currently supports browsing of phenotypes, alleles and stocks as well as querying of sequence features (genes, UniProt matches, InterPro domains, Gene Ontology terms) and of genetic data (phenotypes, stocks, RNA interference experiments). Forms allow submission of RNA interference data and some bioinformatics services are available. Future ParameciumDB development plans include coordination of human curation of the near 40 000 gene models by members of the research community.

  18. Growth of Paramecium tetraurelia in bacterized, monoxenic cultures.

    PubMed

    Enright, W J; Hennessey, T M

    1987-05-01

    Wild type and mutant Paramecium tetraurelia were grown in monoxenic cultures by first growing Enterobacter aerogenes on a defined medium and then adding the Paramecium to the stationary phase bacterial culture. The bacterial growth was proportional to the concentration of the carbon source (citrate), and the Paramecium growth was dependent upon both the bacterial density and the starting density of Paramecium. The behavior, electrophysiological properties, ciliary lipid composition, and growth characteristics were similar to the commonly used bacterized medium (Cerophyl) except that 5-10 times greater Paramecium yields were reliably obtained.

  19. Accumulation of DNA damages in aging Paramecium tetraurelia.

    PubMed

    Holmes, G E; Holmes, N R

    1986-07-01

    Paramecium tetraurelia cells of ages 4, 15, and 27 days were labeled with [14C]-thymidine. In addition, cells were grown clonally for 27 days (108 generations) and labeled with [14C]-thymidine in the presence of 0.5 or 7.5 micrograms/ml of mitomycin-C (MMC) or no MMC. These cells were gently deposited on a filter membrane, which impedes the passage of DNA strands. The cells were then lysed with detergents and the cellular components washed through the filters, leaving double-stranded DNA intact on the surface. Proteinase K was used to remove histone or DNA-bound proteins. The DNA was then eluted under alkaline conditions, which denatures double-stranded DNA and converts apurinic/apyrimidinic sites into single-strand breaks. The results obtained with the cells of ages 4, 15, and 27 days (16, 60, and 108 generations, respectively) indicate that as Paramecium tetraurelia ages during asexual reproduction, apurinic/apyrimidinic lesions, strand breaks or single-strand gaps accumulate. This accumulation may be the basic mechanism of aging in such cells. In the MMC-treated cells of 27 days (108 generations), the MMC reduced elution of DNA fragments more at the higher than at the lower pH's used; random MMC cross-links should occur more often in longer strands than in shorter strands. The reductions in elution preferentially at higher pH, at which longer single strands would be eluted, confirmed the pH-versus-length relationship for Paramecium DNA eluted under our conditions.

  20. Orienting Paramecium with intense static magnetic fields

    NASA Astrophysics Data System (ADS)

    Valles, James M., Jr.; Guevorkian, Karine; Quindel, Carl

    2004-03-01

    Recent experiments on cell division suggest the application of intense static magnetic fields as a novel tool for the manipulation of biological systems [1]. The magnetic field appears to couple to the intrinsic anisotropies in the diamagnetic components of the cells. Here, we present measurements of the intrinsic average diamagnetic anisotropy of the whole single celled ciliate, Paramecium Caudatum. Magnetic fields, 2.5 T < B < 8 T were applied to immobilized (non-swimming) Paramecium Caudatum that were suspended in a density matched medium. The organisms align with their long axis parallel to the applied magnetic field. Their intrinsic diamagnetic anisotropy is 3x10-11 in cgs units. We will discuss the implications of these results for employing magnetic fields to probe the behavior of swimming Paramecium. [1] J. M. Valles, Jr. et al., Expt. Cell Res.274, 112-118 (2002).

  1. The mechanics of gravitaxis in Paramecium.

    PubMed

    Roberts, A M

    2010-12-15

    An analysis of swimming patterns in the ciliate Paramecium shows that the ability to swim preferentially upwards (negative gravitaxis) is primarily the result of upwardly curving trajectories. The trajectory characteristics are consistent with those produced by mechanical orientation. Cell profile measurements from microscope images suggest that the characteristic front-rear body asymmetry accounts for the observed orientation rates. Gravikinesis may result from interactions between the propelling cilia and the sedimentary flow around the cell, and it seems unlikely that an internal physiological gravity receptor exists in Paramecium.

  2. Locomotion of Paramecium in patterned environments

    NASA Astrophysics Data System (ADS)

    Park, Eun-Jik; Eddins, Aja; Kim, Junil; Yang, Sung; Jana, Saikat; Jung, Sunghwan

    2011-10-01

    Ciliary organisms like Paramecium Multimicronucleatum locomote by synchronized beating of cilia that produce metachronal waves over their body. In their natural environments they navigate through a variety of environments especially surfaces with different topology. We study the effects of wavy surfaces patterned on the PDMS channels on the locomotive abilities of Paramecium by characterizing different quantities like velocity amplitude and wavelength of the trajectories traced. We compare this result with the swimming characteristics in straight channels and draw conclusions about the effects of various patterned surfaces.

  3. Paramecium swimming in a capillary tube

    NASA Astrophysics Data System (ADS)

    Jana, Saikat; Jung, Sunghwan

    2010-03-01

    Micro-organisms exhibit different strategies for swimming in complex environments. Many micro-swimmers such as paramecium congregate and tend to live near wall. We investigate how paramecium moves in a confined space as compared to its motion in an unbounded fluid. A new theoretical model based on Taylor's sheet is developed, to study such boundary effects. In experiments, paramecia are put inside capillary tubes and their swimming behavior is observed. The data obtained from experiments is used to test the validity of our theoretical model and understand how the cilia influence the locomotion of paramecia in confined geometries.

  4. Mutagenicity of fly ash particles in Paramecium

    SciTech Connect

    Smith-Sonneborn, J.; Palizzi, R.A.; Herr, C.; Fisher, G.L.

    1981-01-09

    Paramecium, a protozoan that ingests nonnutritive particulate matter, was used to determine the mutagenicity of fly ash. Heat treatment inactivated mutagens that require metabolic conversion to their active form but did not destroy all mutagenicity. Extraction of particles with hydrochloric acid, but not dimethyl sulfoxide, removed detectable mutagenic activity.

  5. Roll and Yaw of Paramecium swimming in a viscous fluid

    NASA Astrophysics Data System (ADS)

    Jung, Sunghwan; Jana, Saikat; Giarra, Matt; Vlachos, Pavlos

    2012-11-01

    Many free-swimming microorganisms like ciliates, flagellates, and invertebrates exhibit helical trajectories. In particular, the Paramecium spirally swims along its anterior direction by the beating of cilia. Due to the oblique beating stroke of cilia, the Paramecium rotates along its long axis as it swims forward. Simultaneously, this long axis turns toward the oral groove side. Combined roll and yaw motions of Paramecium result in swimming along a spiral course. Using Particle Image Velocimetry, we measure and quantify the flow field and fluid stress around Paramecium. We will discuss how the non-uniform stress distribution around the body induces this yaw motion.

  6. Orientation of Paramecium under the conditions of weightlessness.

    PubMed

    Hemmersbach-Krause, R; Briegleb, W; Hader, D P; Vogel, K; Grothe, D; Meyer, I

    1993-01-01

    A cell culture of Paramecium with a precise negative gravitaxis was exposed to 4 x l0(-6) g during a parabolic flight of a sounding rocket for 6 min. Computer image analysis revealed that without gravity stimulus the individual swimming paths remained straight. In addition, three reactions could be distinguished. For about 30 s, paramecia maintained the swimming direction they had before onset of low gravity. During the next 20 s, an approximate reversal of the swimming direction occurred. This period was followed by the expected random swimming pattern. Similar behavior was observed under the condition of simulated weightlessness on a fast-rotating clinostat. Control experiments on the ground under hyper-gravity on a low-speed centrifuge microscope and on a vibration test facility proved that the observed effects were caused exclusively by the reduction of gravity.

  7. A theory of gravikinesis in paramecium

    NASA Astrophysics Data System (ADS)

    Machemer, H.

    The archaic eukaryote unicellular microorganism, Paramecium, is propelled by thousands of cilia, which are regulated by modulation of the membrane potential. Ciliates can successfully cope with gravity, which is the phylogenetically oldest stimulus for living things. One mechanism for overcoming sedimentation is negative gravitaxis, an orientational response antiparallel to the gravity vector. We have postulated the existence of a negative gravikinesis in Paramecium, i.e. a modulation of swimming speed as a function of cellular orientation in space. With negative gravikinesis, an upward oriented cell actively augments the rate of forward swimming and depresses active locomotion during downward orientation. A brief outline of the gravikinesis hypothesis is given on a quantitative basis and experimental data are presented which have confirmed the major assumptions.

  8. Effect of confinements: Bending in Paramecium

    NASA Astrophysics Data System (ADS)

    Eddins, Aja; Yang, Sung; Spoon, Corrie; Jung, Sunghwan

    2012-02-01

    Paramecium is a unicellular eukaryote which by coordinated beating of cilia, generates metachronal waves which causes it to execute a helical trajectory. We investigate the swimming parameters of the organism in rectangular PDMS channels and try to quantify its behavior. Surprisingly a swimming Paramecium in certain width of channels executes a bend of its flexible body (and changes its direction of swimming) by generating forces using the cilia. Considering a simple model of beam constrained between two walls, we predict the bent shapes of the organism and the forces it exerts on the walls. Finally we try to explain how bending (by sensing) can occur in channels by conducting experiments in thin film of fluid and drawing analogy to swimming behavior observed in different cases.

  9. Somersault of Paramecium in extremely confined environments

    PubMed Central

    Jana, Saikat; Eddins, Aja; Spoon, Corrie; Jung, Sunghwan

    2015-01-01

    We investigate various swimming modes of Paramecium in geometric confinements and a non-swimming self-bending behavior like a somersault, which is quite different from the previously reported behaviors. We observe that Paramecia execute directional sinusoidal trajectories in thick fluid films, whereas Paramecia meander around a localized region and execute frequent turns due to collisions with adjacent walls in thin fluid films. When Paramecia are further constrained in rectangular channels narrower than the length of the cell body, a fraction of meandering Paramecia buckle their body by pushing on the channel walls. The bucking (self-bending) of the cell body allows the Paramecium to reorient its anterior end and explore a completely new direction in extremely confined spaces. Using force deflection method, we quantify the Young’s modulus of the cell and estimate the swimming and bending powers exerted by Paramecium. The analysis shows that Paramecia can utilize a fraction of its swimming power to execute the self-bending maneuver within the confined channel and no extra power may be required for this new kind of self-bending behavior. This investigation sheds light on how micro-organisms can use the flexibility of the body to actively navigate within confined spaces. PMID:26286234

  10. Somersault of Paramecium in extremely confined environments

    NASA Astrophysics Data System (ADS)

    Jana, Saikat; Eddins, Aja; Spoon, Corrie; Jung, Sunghwan

    2015-08-01

    We investigate various swimming modes of Paramecium in geometric confinements and a non-swimming self-bending behavior like a somersault, which is quite different from the previously reported behaviors. We observe that Paramecia execute directional sinusoidal trajectories in thick fluid films, whereas Paramecia meander around a localized region and execute frequent turns due to collisions with adjacent walls in thin fluid films. When Paramecia are further constrained in rectangular channels narrower than the length of the cell body, a fraction of meandering Paramecia buckle their body by pushing on the channel walls. The bucking (self-bending) of the cell body allows the Paramecium to reorient its anterior end and explore a completely new direction in extremely confined spaces. Using force deflection method, we quantify the Young’s modulus of the cell and estimate the swimming and bending powers exerted by Paramecium. The analysis shows that Paramecia can utilize a fraction of its swimming power to execute the self-bending maneuver within the confined channel and no extra power may be required for this new kind of self-bending behavior. This investigation sheds light on how micro-organisms can use the flexibility of the body to actively navigate within confined spaces.

  11. Somersault of Paramecium in extremely confined environments.

    PubMed

    Jana, Saikat; Eddins, Aja; Spoon, Corrie; Jung, Sunghwan

    2015-08-19

    We investigate various swimming modes of Paramecium in geometric confinements and a non-swimming self-bending behavior like a somersault, which is quite different from the previously reported behaviors. We observe that Paramecia execute directional sinusoidal trajectories in thick fluid films, whereas Paramecia meander around a localized region and execute frequent turns due to collisions with adjacent walls in thin fluid films. When Paramecia are further constrained in rectangular channels narrower than the length of the cell body, a fraction of meandering Paramecia buckle their body by pushing on the channel walls. The bucking (self-bending) of the cell body allows the Paramecium to reorient its anterior end and explore a completely new direction in extremely confined spaces. Using force deflection method, we quantify the Young's modulus of the cell and estimate the swimming and bending powers exerted by Paramecium. The analysis shows that Paramecia can utilize a fraction of its swimming power to execute the self-bending maneuver within the confined channel and no extra power may be required for this new kind of self-bending behavior. This investigation sheds light on how micro-organisms can use the flexibility of the body to actively navigate within confined spaces.

  12. Association of Paramecium bursaria Chlorella viruses with Paramecium bursaria cells: ultrastructural studies.

    PubMed

    Yashchenko, Varvara V; Gavrilova, Olga V; Rautian, Maria S; Jakobsen, Kjetill S

    2012-05-01

    Paramecium bursaria Chlorella viruses were observed by applying transmission electron microscopy in the native symbiotic system Paramecium bursaria (Ciliophora, Oligohymenophorea) and the green algae Chlorella (Chlorellaceae, Trebouxiophyceae). Virus particles were abundant and localized in the ciliary pits of the cortex and in the buccal cavity of P. bursaria. This was shown for two types of the symbiotic systems associated with two types of Chlorella viruses - Pbi or NC64A. A novel quantitative stereological approach was applied to test whether virus particles were distributed randomly on the Paramecium surface or preferentially occupied certain zones. The ability of the virus to form an association with the ciliate was investigated experimentally; virus particles were mixed with P. bursaria or with symbiont-free species P. caudatum. Our results confirmed that in the freshwater ecosystems two types of P. bursaria -Chlorella symbiotic systems exist, those without Chlorella viruses and those associated with a large amount of the viruses. The fate of Chlorella virus particles at the Paramecium surface was determined based on obtained statistical data and taking into account ciliate feeding currents and cortical reorganization during cell division. A life cycle of the viruses in the complete symbiotic system is proposed. Copyright © 2011 Elsevier GmbH. All rights reserved.

  13. Secondary structural analyses of ITS1 in Paramecium.

    PubMed

    Hoshina, Ryo

    2010-01-01

    The nuclear ribosomal RNA gene operon is interrupted by internal transcribed spacer (ITS) 1 and ITS2. Although the secondary structure of ITS2 has been widely investigated, less is known about ITS1 and its structure. In this study, the secondary structure of ITS1 sequences for Paramecium and other ciliates was predicted. Each Paramecium ITS1 forms an open loop with three helices, A through C. Helix B was highly conserved among Paramecium, and similar helices were found in other ciliates. A phylogenetic analysis using the ITS1 sequences showed high-resolution, implying that ITS1 is a good tool for species-level analyses.

  14. Use of a Novel Cell Adhesion Method and Digital Measurement to Show Stimulus-dependent Variation in Somatic and Oral Ciliary Beat Frequency in Paramecium

    PubMed Central

    Bell, Wade E.; Hallworth, Richard; Wyatt, Todd A.; Sisson, Joseph H.

    2015-01-01

    When Paramecium encounters positive stimuli, the membrane hyperpolarizes and ciliary beat frequency increases. We adapted an established immobilization protocol using a biological adhesive and a novel digital analysis system to quantify beat frequency in immobilized Paramecium. Cells showed low mortality and demonstrated beat frequencies consistent with previous studies. Chemoattractant molecules, reduction in external potassium, and posterior stimulation all increased somatic beat frequency. In all cases, the oral groove cilia maintained a higher beat frequency than mid-body cilia, but only oral cilia from cells stimulated with chemoattactants showed an increase from basal levels. PMID:25066640

  15. Use of a novel cell adhesion method and digital measurement to show stimulus-dependent variation in somatic and oral ciliary beat frequency in Paramecium.

    PubMed

    Bell, Wade E; Hallworth, Richard; Wyatt, Todd A; Sisson, Joseph H

    2015-01-01

    When Paramecium encounters positive stimuli, the membrane hyperpolarizes and ciliary beat frequency increases. We adapted an established immobilization protocol using a biological adhesive and a novel digital analysis system to quantify beat frequency in immobilized Paramecium. Cells showed low mortality and demonstrated beat frequencies consistent with previous studies. Chemoattractant molecules, reduction in external potassium, and posterior stimulation all increased somatic beat frequency. In all cases, the oral groove cilia maintained a higher beat frequency than mid-body cilia, but only oral cilia from cells stimulated with chemoattactants showed an increase from basal levels. © 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.

  16. Molecular phylogenetics of representative Paramecium species.

    PubMed

    Maciejewska, Agnieszka

    2007-01-01

    The genus Paramecium has been known to science for 250 years and contains some of the most widely studied species of ciliates. At present, the basic research object for phylogenetic studies is the genome of various paramecia. One of the most widely used markers are genes coding for various rRNA's. Comparative analyses of sequences coding rRNA were applied for resolving the systematic position of some paramecia species and also for the establishment of an accurate taxonomy of Paramecium. Paramecia were also model organisms for their systematic group in more general studies in a comparative analysis among ciliates, fungi, plants and multicellular animals, illustrating the evolutionary relationships between Archaebacteria and Eucaryota. A new, revolutionary genealogy proposed the shifting of presumptively advanced groups towards more primitive ones, and traditionally primitive forms were located closer to highly specialized taxa, but rRNA analysis did not unambiguously resolve associations within the studied groups. Because of the aforementioned concerns, the number of molecular markers used for alternative studies is growing, such as genes coding proteins from the Hsp family or histone proteins. Other promising candidate markers may be hemoglobin genes or genes coding á-tubulins. In case of comparative analyses ofnucleotide sequences, the outcome of the research usually depends upon a subjective choice of DNA. One of the directions of research in molecular phylogenetics include indirect methods that allow for an estimation of entire genomes, for example RAPD-PCR-fingerprinting.

  17. Malic dehydrogenase locus of Paramecium tetraurelia.

    PubMed

    Williams, T J; Smith-Sonneborn, J

    1980-04-01

    A search was undertaken for naturally occurring genetic markers for use in clonal aging studies of Paramecium tetraurelia. Clonal age is defined as the number of cell divisions since the last sexual process. Autogamy (self-fertilization) is a sexual process which can occur in aging lines, resulting in homozygosity and initiation of the next generation. Such "illicit" autogamies must be detected and eliminated from the aged clone. With codominant alleles, heterozygous aging lines can be established which will express a phenotype distinguishable from that of either parental type and autogamy can then be monitored by the appearance of either segregant homozygous phenotype. However, very few codominant alleles are available in this species. Electrophoretic mobilities of malic dehydrogenase (MDH) were assayed in 11 stocks of Paramecium tetraurelia by polyacrylamide gel electrophoresis. Nine stocks showed a single-banded "stock 51" type, while stock 174 and stock 29 each exhibited unique mobility. Crosses between stock 51 and the deviant stocks revealed distinct three-banded patterns indicative of heterozygosity of the F1 generation. In the autogamous F2 generation, 1:1 segregation of the parental types were recovered. The pattern of inheritance is consistent with codominant alleles and Mendelian inheritance. These naturally occurring biochemical markers are stable with increasing clonal age and are therefore useful genetic markers for studies of cellular aging.

  18. Methods for Studying Ciliary-Mediated Chemoresponse in Paramecium.

    PubMed

    Valentine, Megan Smith; Van Houten, Judith L

    2016-01-01

    Paramecium is a useful model organism for the study of ciliary-mediated chemical sensing and response. Here we describe ways to take advantage of Paramecium to study chemoresponse.Unicellular organisms like the ciliated protozoan Paramecium sense and respond to chemicals in their environment (Van Houten, Ann Rev Physiol 54:639-663, 1992; Van Houten, Trends Neurosci 17:62-71, 1994). A thousand or more cilia that cover Paramecium cells serve as antennae for chemical signals, similar to ciliary function in a large variety of metazoan cell types that have primary or motile cilia (Berbari et al., Curr Biol 19(13):R526-R535, 2009; Singla V, Reiter J, Science 313:629-633, 2006). The Paramecium cilia also produce the motor output of the detection of chemical cues by controlling swimming behavior. Therefore, in Paramecium the cilia serve multiple roles of detection and response.We present this chapter in three sections to describe the methods for (1) assaying populations of cells for their behavioral responses to chemicals (attraction and repulsion), (2) characterization of the chemoreceptors and associated channels of the cilia using proteomics and binding assays, and (3) electrophysiological analysis of individual cells' responses to chemicals. These methods are applied to wild type cells, mutants, transformed cells that express tagged proteins, and cells depleted of gene products by RNA Interference (RNAi).

  19. Exploration of Self-Regulation in the Natural Swimming of the Paramecium’s Cilium

    DTIC Science & Technology

    2012-02-01

    aspects of bio-physical self-regulation of this motion) in the paramecium’s cilium. The paramecium is a single-cell animal widely found in oxygenated...of Ciliary Motion Hardness Control Natural Swimming Paramecium Robotic Cilium Underwater Acoustic Transduction 16. SECURITY CLASSIFICATION OF: a...Comparison of the Model with Measurements Due to Machemer (1972) 6 2 Comparison of the Tracks of the Cilium of a Paramecium as Measured by Machemer

  20. Bioconvection and front formation of Paramecium tetraurelia

    NASA Astrophysics Data System (ADS)

    Kitsunezaki, So; Komori, Rie; Harumoto, Terue

    2007-10-01

    We have investigated the bioconvection of Paramecium tetraurelia in high-density suspensions made by centrifugal concentration. When a suspension is kept at rest in a Hele-Shaw cell, a crowded front of paramecia is formed in the vicinity of the bottom and it propagates gradually toward the water-air interface. Fluid convection occurs under this front, and it is driven persistently by the upward swimming of paramecia. The roll structures of the bioconvection become turbulent with an increase in the depth of the suspension; they also change rapidly as the density of paramecia increases. Our experimental results suggest that lack of oxygen in the suspension causes the active individual motions of paramecia to induce the formation of this front.

  1. Calmodulin antagonists inhibit secretion in Paramecium

    PubMed Central

    1983-01-01

    Secretion in Paramecium is Ca2+-dependent and involves exocytic release of the content of the secretory organelle, known as the trichocyst. The content, called the trichocyst matrix, undergoes a Ca2+-induced reordering of its paracrystalline structure during release, and we have defined three stages in this expansion process. The stage I, or fully condensed trichocyst, is the 4 microns-long membrane-bounded form existing prior to stimulation. Stage II, the partially expanded trichocyst, we define as an intermediate stage in the transition, preceding stage III, the fully expanded extruded form which is a 20-40 microns-long needlelike structure. These stages have been used to assay the effects of trifluoperazine (TFP) and W-7, calmodulin (CaM) antagonists, on trichocyst matrix expansion in vivo. TFP and W-7 are shown to reversibly block matrix release induced by picric acid. Ultra- structural examination reveals that one effect of this inhibition is reflected in the organelles themselves, which are prevented from undergoing the stage I-stage II transition by preincubation in 14 microM TFP or 35 microM W-7 before fixation. This inhibition of expansion by TFP can be moderated but not abolished by high extracellular Ca2+ (5 mM). The moderation by high Ca2+ can be eliminated by raising TFP concentration to 20 microM. A possible explanation for the ability to titrate the inhibition in this manner is that TFP is acting to block expansion by binding to the Ca2+-CaM complex. Brief exposure of cells to the Ca2+ ionophore A23187 and 5 mM Ca2+ following TFP treatment promotes matrix expansion, although in 14 microM TFP a residual level of inhibition remains. These results suggest that, following stimulation, CaM regulates secretion in Paramecium, possibly by controlling the Ca2+-dependent matrix expansion which accompanies exocytosis in these cells. PMID:6403556

  2. [Comparative description of macronuclear electrophoretic karyotypes of Paramecium primaurelia and Paramecium novaurelia sibling species].

    PubMed

    Nekrasova, I V; Potekhin, A A; Przybos, E; Rautian, M S

    2008-01-01

    The macronuclear genomes of two sibling species belonging to the Paramecium aurelia complex--P. primaurelia and P. novaurelia--were analyzed by pulsed-field gel electrophoresis (PFGE). Their electrophoretic karyotypes showed a continuous spectrum of different-sized DNA molecules with a species-specific pattern of banding, which was reproducible and did not change with strain senescence. Thus, P. aurelia macronuclear PFGE profiles could be described by an approach analogous to that used for the description of metaphase chromosome banding patterns. At first, well-identifiable regions (size fractions) of a PFGE profile, which can be seen at any resolution, are determined. Then, the bands of the second order of resolution (subfractions) can be defined in some of these regions. The blocks of the first and second orders can be utilized as inner markers of the PFGE profile allowing precise comparison of different PFGE profiles. Such comparative analysis has demonstrated some marking differences in organization of the macronuclear genomes of P. primaurelia and P. novaurelia, and low level of intraspecific polymorphism. Worth noting is that the P. novaurelia strain isolated in USA was different from all other analyzed P. novaurelia strains originating from Europe. The nature of banding of P. aurelia PFGE profiles is discussed. The revealed high order and stability of the macronuclear genome organization makes possible searching for new approaches to study the mechanisms of this non-trivial genome formation and maintenance. Further PFGE analysis of the macronuclear genomes of the other Paramecium species in relation with the Paramecium phylogeny may provide insights into the directions of the evolution of the macronuclear genome in Ciliata.

  3. Simulation of Paramecium Chemotaxis Exposed to Calcium Gradients.

    PubMed

    Sarvestani, Ali N; Shamloo, Amir; Ahmadian, Mohammad Taghi

    2016-06-01

    Paramecium or other ciliates have the potential to be utilized for minimally invasive surgery systems, making internal body organs accessible. Paramecium shows interesting responses to changes in the concentration of specific ions such as K(+), Mg(2+), and Ca(2+) in the ambient fluid. Some specific responses are observed as, changes in beat pattern of cilia and swimming toward or apart from the ion source. Therefore developing a model for chemotactic motility of small organisms is necessary in order to control the directional movements of these microorganisms before testing them. In this article, we have developed a numerical model, investigating the effects of Ca(2+) on swimming trajectory of Paramecium. Results for Ca(2+)-dependent chemotactic motility show that calcium gradients are efficient actuators for controlling the Paramecium swimming trajectory. After applying a very low Ca(2+) gradient, a directional chemotaxis of swimming Paramecium is observable in this model. As a result, chemotaxis is shown to be an efficient method for controlling the propulsion of these small organisms.

  4. Survey of Paramecium duboscqui using three markers and assessment of the molecular variability in the genus Paramecium.

    PubMed

    Boscaro, Vittorio; Fokin, Sergei I; Verni, Franco; Petroni, Giulio

    2012-12-01

    The genus Paramecium (phylum Ciliophora) is one of the best-known among protozoa. Nevertheless, the knowledge on the diversity and distribution of species within this genus was remarkably scarce until recent times. In the last years a constantly growing amount of data has formed, especially on the distribution of species and the characterization of molecular markers. Much effort has been made on detecting clades inside each morphospecies, which could suggest the presence of sibling species complexes as in the famous case of Paramecium aurelia. In this work we present new data on Paramecium duboscqui, one of the morphospecies that have not yet been surveyed employing DNA sequences as markers. We obtained data from nine strains sampled around the world, using the three most commonly employed markers (18S rRNA gene, ITS1-5.8S-ITS2 and COI gene sequences). Moreover, we compared our results with those already available for other Paramecium species, and performed phylogenetic analyses for the entire genus. We also expanded the knowledge on the ITS2 secondary structure and its usefulness in studies on Paramecium. Our approach, that considers the data of all the species together, highlighted some characteristic patterns as well as some ambiguities that should be further investigated. Copyright © 2012 Elsevier Inc. All rights reserved.

  5. Cell proliferation of Paramecium tetraurelia on a slow rotating clinostat

    NASA Astrophysics Data System (ADS)

    Sawai, Satoe; Mogami, Yoshihiro; Baba, Shoji A.

    Paramecium is known to proliferate faster under microgravity conditions, and slower under hypergravity. Experiments using axenic culture medium have demonstrated that hypergravity affected directly on the proliferation of Paramecium itself. In order to assess the mechanisms underlying the physiological effects of gravity on cell proliferation, Paramecium tetraurelia was grown under clinorotation (2.5 rpm) and the time course of the proliferation was investigated in detail on the basis of the logistic analysis. On the basis of the mechanical properties of Paramecium, this slow rate of the rotation appears to be enough to simulate microgravity in terms of the randomization of the cell orientation with respect to gravity. P. tetraurelia was cultivated in a closed chamber in which cells were confined without air bubbles, reducing the shear forces and turbulences under clinorotation. The chamber is made of quartz and silicone rubber film; the former is for the optically-flat walls for the measurement of cell density by means of a non-invasive laser optical-slice method, and the latter for gas exchange. Because of the small dimension for culture space, Paramecium does not accumulate at the top of the chamber in spite of its known negative gravitactic behavior. We measured the cell density at regular time intervals without breaking the configuration of the chamber, and analyzed the proliferation parameters by fitting the data to a logistic equation. As a result, P. tetraurelia showed reduced proliferation under slow clinorotation. The saturation of the cell density as well as the maximum proliferation rate decreased, although we found no significant changes on the half maximal time for proliferation. We also found that the mean swimming velocity decreased under slow clinorotation. These results were not consistent with those under microgravity and fast rotating clinostat. This may suggest that randomization of the cell orientation performed by slow rotating clinostat has

  6. Pb2+ Modulates Ca2+ Membrane Permeability In Paramecium

    NASA Astrophysics Data System (ADS)

    Bernal-Martínez, Juan; Ortega Soto, Arturo

    2004-09-01

    Intracellular recording experiments in current clamp configuration were done to evaluate whether Pb2+ modulates ionic membrane permeability in the fresh water Paramecium tetraurelia. It was found that Pb2+ triggers in a dose-dependent manner, a burst of spontaneous action potentials followed by a robust and sustained after hyper-polarization. In addition, Pb2+ increased the frequency of firing the spontaneous Ca2+-Action Potential and also, the duration of Ca2+-Action Potential, in a dose and reversibly-dependent manner. These results suggest that Pb2+ increases calcium membrane permeability of Paramecium and probably activates a calcium-dependent-potassium conductance in the ciliate.

  7. Cell proliferation of Paramecium tetraurelia under simulated microgravity

    NASA Astrophysics Data System (ADS)

    Sawai, S.; Mogami, Y.; Baba, S. A.

    Paramecium is known to proliferate faster under microgravity in space and slower under hypergravity Experiments using axenic culture medium have demonstrated that the hypergravity affected directly on the proliferation of Paramecium itself Kato et al 2003 In order to assess the mechanisms underlying the physiological effects of gravity on cell proliferation Paramecium tetraurelia was grown under simulated microgravity performed by clinorotation and the time course of the proliferation was investigated in detail on the basis of the logistic analysis P tetraurelia was cultivated in a closed chamber in which cells were confined without air babbles reducing the shear stresses and turbulence under the rotation The chamber is made of quartz and silicone rubber film the former is for the optically-flat walls for the measurement of cell density by means of a non-invasive laser optical-slice method and the latter for gas exchange Because the closed chamber has an inner dimension of 3 times 3 times 60 mm Paramecium does not accumulate at the top of the chamber despite its negative gravitactic behavior We measured the cell density at regular time intervals without breaking the configuration of the chamber and analyzed the proliferation parameters by fitting the data to a logistic equation Clinorotation had the effects of reducing the proliferation of P tetraurelia It reduced both the saturation cell density and the maximum proliferation rate although it had little effect on the

  8. Remembrance of things past retrieved from the Paramecium genome.

    PubMed

    Sperling, Linda

    2011-01-01

    Paramecium and other ciliates are the only unicellular eukaryotes that separate germinal and somatic functions. A germline micronucleus transmits the genetic information to sexual progeny, while a somatic macronucleus expresses the genetic information during vegetative growth to determine the phenotype. At each sexual generation, a new macronucleus develops from the zygotic nucleus through programmed rearrangements of the germline genome. Paramecium tetraurelia somatic genome sequencing, reviewed here, has provided insight into the organization and evolution of the genome. A series of at least 3 whole genome duplications was detected in the Paramecium lineage and selective pressures that determine the fate of the gene duplicates analyzed. Variability in the somatic DNA was characterized and could be attributed to the genome rearrangement processes. Since, in Paramecium, alternative genome rearrangement patterns can be inherited across sexual generations by homology-dependent epigenetic mechanisms and can affect phenotype, I discuss the possibility that ciliate nuclear dimorphism buffers genetic variation hidden in the germline. Copyright © 2011 Institut Pasteur. Published by Elsevier SAS. All rights reserved.

  9. Calcium regulation in the protozoan model, Paramecium tetraurelia.

    PubMed

    Plattner, Helmut

    2014-01-01

    Early in eukaryotic evolution, the cell has evolved a considerable inventory of proteins engaged in the regulation of intracellular Ca(2+) concentrations, not only to avoid toxic effects but beyond that to exploit the signaling capacity of Ca(2+) by small changes in local concentration. Among protozoa, the ciliate Paramecium may now be one of the best analyzed models. Ciliary activity and exo-/endocytosis are governed by Ca(2+) , the latter by Ca(2+) mobilization from alveolar sacs and a superimposed store-operated Ca(2+) -influx. Paramecium cells possess plasma membrane- and endoplasmic reticulum-resident Ca(2+) -ATPases/pumps (PMCA, SERCA), a variety of Ca(2+) influx channels, including mechanosensitive and voltage-dependent channels in the plasma membrane, furthermore a plethora of Ca(2+) -release channels (CRC) of the inositol 1,4,5-trisphosphate and ryanodine receptor type in different compartments, notably the contractile vacuole complex and the alveolar sacs, as well as in vesicles participating in vesicular trafficking. Additional types of CRC probably also occur but they have not been identified at a molecular level as yet, as is the equivalent of synaptotagmin as a Ca(2+) sensor for exocytosis. Among established targets and sensors of Ca(2+) in Paramecium are calmodulin, calcineurin, as well as Ca(2+) /calmodulin-dependent protein kinases, all with multiple functions. Thus, basic elements of Ca(2+) signaling are available for Paramecium. © 2013 The Author(s) Journal of Eukaryotic Microbiology © 2013 International Society of Protistologists.

  10. Secondary symbiosis between Paramecium and Chlorella cells.

    PubMed

    Kodama, Yuuki; Fujishima, Masahiro

    2010-01-01

    Each symbiotic Chlorella species of Paramecium bursaria is enclosed in a perialgal vacuole (PV) membrane derived from the host digestive vacuole (DV) membrane. Algae-free paramecia and symbiotic algae are capable of growing independently and paramecia can be reinfected experimentally by mixing them. This phenomenon provides an excellent model for studying cell-to-cell interaction and the evolution of eukaryotic cells through secondary endosymbiosis between different protists. However, the detailed algal infection process remains unclear. Using pulse labeling of the algae-free paramecia with the isolated symbiotic algae and chase method, we found four necessary cytological events for establishing endosymbiosis. (1) At about 3 min after mixing, some algae show resistance to the host lysosomal enzymes in the DVs, even if the digested ones are present. (2) At about 30 min after mixing, the alga starts to escape from the DVs as the result of the budding of the DV membrane into the cytoplasm. (3) Within 15 min after the escape, the DV membrane enclosing a single green alga differentiates to the PV membrane, which provides protection from lysosomal fusion. (4) The alga localizes at the primary lysosome-less host cell surface by affinity of the PV to unknown structures of the host. At about 24 h after mixing, the alga multiplies by cell division and establishes endosymbiosis. Infection experiments with infection-capable and infection-incapable algae indicate that the infectivity of algae is based on their ability to localize beneath the host surface after escaping from the DVs. This algal infection process differs from known infection processes of other symbiotic or parasitic organisms to their hosts. Copyright 2010 Elsevier Inc. All rights reserved.

  11. Graviresponses of Paramecium biaurelia during parabolic flights.

    PubMed

    Krause, Martin; Bräucker, Richard; Hemmersbach, Ruth

    2006-12-01

    The thresholds of graviorientation and gravikinesis in Paramecium biaurelia were investigated during the 5th DLR (German Aerospace Center) parabolic-flight campaign at Bordeaux in June 2003. Parabolic flights are a useful tool for the investigation of swimming behaviour in protists at different accelerations. At normal gravity (1 g) and hypergravity (1 g to 1.8 g), precision of orientation and locomotion rates depend linearly on the applied acceleration as seen in earlier centrifuge experiments. After transition from hypergravity to decreased gravity (minimal residual acceleration of <10(-2) g), graviorientation as well as gravikinesis show a full relaxation with different kinetics. The use of twelve independent cell samples per flight guarantees high data numbers and secures the statistical significance of the obtained data. The relatively slow change of acceleration between periods of microgravity and hypergravity (0.4 g/s) enabled us to determine the thresholds of graviorientation at 0.6 g and of gravikinesis at 0.4 g. The gravity-unrelated propulsion rate of the sample was found to be 874 microm/s, exceeding the locomotion rate of horizontally swimming cells (855 microm/s). The measured thresholds of graviresponses were compared with data obtained from earlier centrifuge experiments on the sounding rocket Maxus-2. Measured thresholds of gravireactions indicate that small energies, close to the thermal noise level, are sufficient for the gravitransduction process. Data from earlier hypergravity experiments demonstrate that mechanosensitive ion channels are functioning over a relative wide range of acceleration. From this, we may speculate that gravireceptor channels derive from mechanoreceptor channels.

  12. Detecting the gravitational sensitivity of Paramecium caudatum using magnetic forces

    NASA Astrophysics Data System (ADS)

    Guevorkian, Karine; Valles, James M., Jr.

    2006-03-01

    Under normal conditions, Paramecium cells regulate their swimming speed in response to the pN level mechanical force of gravity. This regulation, known as gravikinesis, is more pronounced when the external force is increased by methods such as centrifugation. Here we present a novel technique that simulates gravity fields using the interactions between strong inhomogeneous magnetic fields and cells. We are able to achieve variable gravities spanning from 10xg to -8xg; where g is earth's gravity. Our experiments show that the swimming speed regulation of Paramecium caudatum to magnetically simulated gravity is a true physiological response. In addition, they reveal a maximum propulsion force for paramecia. This advance establishes a general technique for applying continuously variable forces to cells or cell populations suitable for exploring their force transduction mechanisms.

  13. Genetic Diversity in the Paramecium aurelia Species Complex

    PubMed Central

    Catania, Francesco; Wurmser, François; Potekhin, Alexey A.; Przyboś, Ewa; Lynch, Michael

    2009-01-01

    Current understanding of the population genetics of free-living unicellular eukaryotes is limited, and the amount of genetic variability in these organisms is still a matter of debate. We characterized—reproductively and genetically—worldwide samples of multiple Paramecium species belonging to a cryptic species complex, Paramecium aurelia, whose species have been shown to be reproductively isolated. We found that levels of genetic diversity both in the nucleus and in the mitochondrion are substantial within groups of reproductively compatible P. aurelia strains but drop considerably when strains are partitioned according to their phylogenetic groupings. Our study reveals the existence of discrepancies between the mating behavior of a number of P. aurelia strains and their multilocus genetic profile, a controversial finding that has major consequences for both the current methods of species assignment and the species problem in the P. aurelia complex. PMID:19023087

  14. Transitions between three swimming gaits in Paramecium escape.

    PubMed

    Hamel, Amandine; Fisch, Cathy; Combettes, Laurent; Dupuis-Williams, Pascale; Baroud, Charles N

    2011-05-03

    Paramecium and other protists are able to swim at velocities reaching several times their body size per second by beating their cilia in an organized fashion. The cilia beat in an asymmetric stroke, which breaks the time reversal symmetry of small scale flows. Here we show that Paramecium uses three different swimming gaits to escape from an aggression, applied in the form of a focused laser heating. For a weak aggression, normal swimming is sufficient and produces a steady swimming velocity. As the heating amplitude is increased, a higher acceleration and faster swimming are achieved through synchronized beating of the cilia, which begin by producing oscillating swimming velocities and later give way to the usual gait. Finally, escape from a life-threatening aggression is achieved by a "jumping" gait, which does not rely on the cilia but is achieved through the explosive release of a group of trichocysts in the direction of the hot spot. Measurements through high-speed video explain the role of trichocysts in defending against aggressions while showing unexpected transitions in the swimming of microorganisms. These measurements also demonstrate that Paramecium optimizes its escape pattern by taking advantage of its inertia.

  15. Transitions between three swimming gaits in Paramecium escape

    PubMed Central

    Hamel, Amandine; Fisch, Cathy; Combettes, Laurent; Dupuis-Williams, Pascale; Baroud, Charles N.

    2011-01-01

    Paramecium and other protists are able to swim at velocities reaching several times their body size per second by beating their cilia in an organized fashion. The cilia beat in an asymmetric stroke, which breaks the time reversal symmetry of small scale flows. Here we show that Paramecium uses three different swimming gaits to escape from an aggression, applied in the form of a focused laser heating. For a weak aggression, normal swimming is sufficient and produces a steady swimming velocity. As the heating amplitude is increased, a higher acceleration and faster swimming are achieved through synchronized beating of the cilia, which begin by producing oscillating swimming velocities and later give way to the usual gait. Finally, escape from a life-threatening aggression is achieved by a “jumping” gait, which does not rely on the cilia but is achieved through the explosive release of a group of trichocysts in the direction of the hot spot. Measurements through high-speed video explain the role of trichocysts in defending against aggressions while showing unexpected transitions in the swimming of microorganisms. These measurements also demonstrate that Paramecium optimizes its escape pattern by taking advantage of its inertia. PMID:21464291

  16. Ciliary heterogeneity within a single cell: the Paramecium model.

    PubMed

    Aubusson-Fleury, Anne; Cohen, Jean; Lemullois, Michel

    2015-01-01

    Paramecium is a single cell able to divide in its morphologically differentiated stage that has many cilia anchored at its cell surface. Many thousands of cilia are thus assembled in a short period of time during division to duplicate the cell pattern while the cell continues swimming. Most, but not all, of these sensory cilia are motile and involved in two main functions: prey capture and cell locomotion. These cilia display heterogeneity, both in their length and their biochemical properties. Thanks to these properties, as well as to the availability of many postgenomic tools and the possibility to follow the regrowth of cilia after deciliation, Paramecium offers a nice opportunity to study the assembly of the cilia, as well as the genesis of their diversity within a single cell. In this paper, after a brief survey of Paramecium morphology and cilia properties, we describe the tools and the protocols currently used for immunofluorescence, transmission electron microscopy, and ultrastructural immunocytochemistry to analyze cilia, with special recommendations to overcome the problem raised by cilium diversity. Copyright © 2015. Published by Elsevier Inc.

  17. A two-locus molecular characterization of Paramecium calkinsi.

    PubMed

    Przyboś, Ewa; Tarcz, Sebastian; Potekhin, Alexey; Rautian, Maria; Prajer, Małgorzata

    2012-03-01

    Paramecium calkinsi (Ciliophora, Protozoa) is a euryhaline species which was first identified in freshwater habitats, but subsequently several strains were also collected from brackish water. It is characterized by clockwise spiral swimming movement and the general morphology of the "bursaria type." The present paper is the first molecular characterization of P. calkinsi strains recently collected in distant regions in Russia using ITS1-5.8S- ITS2-5'LSU rDNA (1100bp) and COI (620bp) mtDNA sequenced gene fragments. For comparison, our molecular analysis includes P. bursaria, exhibiting a similar "bursaria morphotype" as well as species representing the "aurelia type," i.e., P. caudatum, P. multimicronucleatum, P. jenningsi, and P. schewiakoffi, and some species of the P. aurelia species complex (P. primaurelia, P. tetraurelia, P. sexaurelia, and P. tredecaurelia). We also use data from GenBank concerning other species in the genus Paramecium and Tetrahymena (which used as an outgroup). The division of the genus Paramecium into four subgenera (proposed by Fokin et al. 2004) is clearly presented by the trees. There is a clear separation between P. calkinsi strains collected from different regions (races). Consequently, given the molecular distances between them, it seems that these races may represent different syngens within the species. Copyright © 2011 Elsevier GmbH. All rights reserved.

  18. Effects of perfluorinated amphiphiles on backward swimming in Paramecium caudatum

    SciTech Connect

    Matsubara, Eriko; Harada, Kouji; Inoue, Kayoko; Koizumi, Akio . E-mail: koizumi@pbh.med.kyoto-u.ac.jp

    2006-01-13

    PFOS and PFOA are ubiquitous contaminants in the environment. We investigated the effects of fluorochemicals on calcium currents in Paramecium caudatum using its behavioral changes. Negatively charged amphiphiles prolonged backward swimming (BWS) of Paramecium. PFOS significantly prolonged BWS, while PFOA was less potent (EC{sub 5}: 29.8 {+-} 4.1 and 424.1 {+-} 124.0 {mu}M, respectively). The BWS prolongation was blocked by cadmium, indicating that the cellular calcium conductance had been modified. The positively charged amphiphile FOSAPrTMA shortened BWS (EC{sub 5}: 19.1 {+-} 17.3). Nonionic amphiphiles did not affect BWS. The longer-chain perfluorinated carboxylates PFNA and PFDA were more potent than PFOA (EC{sub 5}: 98.7 {+-} 20.1 and 60.4 {+-} 10.1 {mu}M, respectively). However, 1,8-perfluorooctanedioic acid and 1,10-perfluorodecanedioic acid did not prolong BWS. The critical micelle concentration (CMC) and BWS prolongation for negatively charged amphiphiles showed a clear correlation (r {sup 2} = 0.8008, p < 0.001). In summary, several perfluorochemicals and PFOS and PFOA had similar effects in Paramecium, while chain length, CMC, and electric charge were major determinants of BWS duration.

  19. Fine oral filaments in Paramecium: a biochemical and immunological analysis.

    PubMed

    Clerot, J; Iftode, F; Budin, K; Jeanmaire-Wolf, R; Coffe, G; Fleury-Aubusson, A

    2001-01-01

    In Paramecium, several kinds of the oral networks of fine filaments are defined at the ultrastructural level. Using the sodium chloride-treated oral apparatus of Paramecium as an antigen to produce monoclonal antibodies, we have begun to identify the proteins constituting these networks. Immunoblotting showed that all positive antibodies were directed against three bands (70-, 75-and 83-kD), which corresponded to quantitatively minor components of the antigen; there was no antibody specific for the quantitatively major components (58- and 62-kD). Immunolocalization with four of these antibodies directed against one or several of these three bands showed that these proteins are components of the fine filaments supporting the oral area; a decoration of the basal bodies and the outer lattice was also observed on the cortex. Immunofluorescence on interphase cells suggested that the three proteins colocalized on the left side of the oral apparatus, whereas only the 70-kD band was detected on the right side. During division, the antigens of the antibodies were detected at different stages after oral basal body assembly. The antibodies cross-reacted with the tetrins, which are oral filament-forming proteins in Tetrahymena, demonstrating that tetrin-related proteins are quantitatively minor components of the oral and the somatic cytoskeleton of Paramecium.

  20. Species of the Paramecium aurelia complex in Russia: new stands and overall distribution.

    PubMed

    Potekhin, Alexey; Przyboś, Ewa; Nekrasova, Irina; Yashchenko, Varvara; Rautian, Maria

    2010-01-01

    New stands of Paramecium biaurelia, P. triaurelia, P. tetraurelia, P. pentaurelia, P. novaurelia, and P. dodecaurelia were recorded in Russia. Especially interesting is the record of P. novaurelia in Vladivostok, Russian Far East, as it is a very rare species outside of Europe. The distribution of species of the Paramecium aurelia complex in Eurasia with emphasis on findings in Russia is discussed.

  1. Species Identity of Commercial Stocks of Paramecium in the U.S.

    ERIC Educational Resources Information Center

    Cole, Thomas A.; And Others

    1992-01-01

    Describes how paramecium can be identified through the use of DNA-binding fluorescent compounds. The authors used these techniques to test the paramecium stocks from 12 commercial sources. The details of the staining procedures and the results of the commercial tests are presented in this article. (PR)

  2. Paramecium swimming and ciliary beating patterns: a study on four RNA interference mutations.

    PubMed

    Funfak, Anette; Fisch, Cathy; Abdel Motaal, Hatem T; Diener, Julien; Combettes, Laurent; Baroud, Charles N; Dupuis-Williams, Pascale

    2015-01-01

    Paramecium cells swim and feed by beating their thousands of cilia in coordinated patterns. The organization of these patterns and its relationship with cell motility has been the subject of a large body of work, particularly as a model for ciliary beating in human organs where similar organization is seen. However the rapid motion of the cells makes quantitative measurements very challenging. Here we provide detailed measurements of the swimming of Paramecium cells from high-speed video at high magnification, as they move in microfluidic channels. An image analysis protocol allows us to decouple the cell movement from the motion of the cilia, thus allowing us to measure the ciliary beat frequency (CBF) and the spatio-temporal organization into metachronal waves along the cell periphery. Two distinct values of the CBF appear at different regions of the cell: most of the cilia beat in the range of 15 to 45 Hz, while the cilia in the peristomal region beat at almost double the frequency. The body and peristomal CBF display a nearly linear relation with the swimming velocity. Moreover the measurements do not display a measurable correlation between the swimming velocity and the metachronal wave velocity on the cell periphery. These measurements are repeated for four RNAi silenced mutants, where proteins specific to the cilia or to their connection to the cell base are depleted. We find that the mutants whose ciliary structure is affected display similar swimming to the control cells albeit with a reduced efficiency, while the mutations that affect the cilia's anchoring to the cell lead to strongly reduced ability to swim. This reduction in motility can be related to a loss of coordination between the ciliary beating in different parts of the cell.

  3. Purification and characterization of calmodulin (lysine 115) N-methyltransferase from Paramecium tetraurelia.

    PubMed

    Pech, L L; Nelson, D L

    1994-03-02

    Calmodulin (lysine 115) N-methyltransferase was purified from the cytosolic fraction of Paramecium tetraurelia by sequential dialysis, cellulose phosphate chromatography, Reactive Red 120 agarose chromatography, and calmodulin-Sepharose affinity chromatography. The enzyme was purified 6800-fold with a 15% yield. SDS-PAGE analysis of the purified enzyme invariably revealed a major protein of 37 kDa that was reproducibly obtained and minor proteins of 35 and 28 kDa that were sometimes obtained in variable yields. The enzyme formed a mixture of mono-, di-, and trimethyllysine residues at lysine 115 of calmodulin in vitro, had a Km for the methyl donor, S-adenosyl methionine (AdoMet), of about 1 microM and a pH optimum of about 7.5. The purified enzyme had an absolute requirement for the reductant DTT for activity, whereas the enzyme in crude fractions did not. The enzyme is a monomer with an estimated molecular mass of 33 kDa. Ca2+, Mg2+, Mn2+, and Ni2+ stimulated calmodulin N-methyltransferase activity but Zn2+ did not. Calmodulin N-methyltransferase was inhibited by its reaction product S-adenosyl homocysteine (SAH), but not by sinefungin and tubercidin. The calmodulin antagonists calmidazolium and mellitin were inhibitory but W7 was not. The enzyme was not stimulated by Triton X-100 nor by NaCl. Only calmodulins with an unmethylated lysine at residue 115, including cam2 calmodulin, were substrates. Histones and calcium-binding proteins from Paramecium other than calmodulin did not act as substrates for the purified calmodulin N-methyltransferase and no other substrates in the cytosolic fraction were observed.

  4. ParameciumDB in 2011: new tools and new data for functional and comparative genomics of the model ciliate Paramecium tetraurelia.

    PubMed

    Arnaiz, Olivier; Sperling, Linda

    2011-01-01

    ParameciumDB is a community model organism database built with the GMOD toolkit to integrate the genome and biology of the ciliate Paramecium tetraurelia. Over the last four years, post-genomic data from proteome and transcriptome studies has been incorporated along with predicted orthologs in 33 species, annotations from the community and publications from the scientific literature. Available tools include BioMart for complex queries, GBrowse2 for genome browsing, the Apollo genome editor for expert curation of gene models, a Blast server, a motif finder, and a wiki for protocols, nomenclature guidelines and other documentation. In-house tools have been developed for ontology browsing and evaluation of off-target RNAi matches. Now ready for next-generation deep sequencing data and the genomes of other Paramecium species, this open-access resource is available at http://paramecium.cgm.cnrs-gif.fr.

  5. Anesthetic action of volatile anesthetics by using Paramecium as a model.

    PubMed

    Zhou, Miaomiao; Xia, Huimin; Xu, Younian; Xin, Naixing; Liu, Jiao; Zhang, Shihai

    2012-06-01

    Although empirically well understood in their clinical administration, volatile anesthetics are not yet well comprehended in their mechanism studies. A major conundrum emerging from these studies is that there is no validated model to assess the presumed candidate sites of the anesthetics. We undertook this study to test the hypothesis that the single-celled Paramecium could be anesthetized and served as a model organism in the study of anesthetics. We assessed the motion of Paramecium cells with Expert Vision system and the chemoresponse of Paramecium cells with T-maze assays in the presence of four different volatile anesthetics, including isoflurane, sevoflurane, enflurane and ether. Each of those volatiles was dissolved in buffers to give drug concentrations equal to 0.8, 1.0, and 1.2 EC50, respectively, in clinical practice. We could see that after application of volatile anesthetics, the swimming of the Paramecium cells was accelerated and then suppressed, or even stopped eventually, and the index of the chemoresponse of the Paramecium cells (denoted as I ( che )) was decreased. All of the above impacts were found in a concentration-dependent fashion. The biphasic effects of the clinical concentrations of volatile anesthetics on Paramecium simulated the situation of high species in anesthesia, and the inhibition of the chemoresponse also indicated anesthetized. In conclusion, the findings in our studies suggested that the single-celled Paramecium could be anesthetized with clinical concentrations of volatile anesthetics and therefore be utilized as a model organism to study the mechanisms of volatile anesthetics.

  6. Longevity of a Paramecium cell clone in space: Hypergravity experiments as a basis for microgravity experiments

    NASA Astrophysics Data System (ADS)

    Kato, Yuko; Mogami, Yoshihiro; Baba, Shoji A.

    We proposed a space experiment aboard International Space Station to explore the effects of microgravity on the longevity of a Paramecium cell clone. Earlier space experiments in CYTOS and Space Lab D-1 demonstrated that Paramecium proliferated faster in space. In combination with the fact that aging process in Paramecium is largely related to the fission age, the results of the proliferation experiment in space may predict that the longevity of Paramecium decreases when measured by clock time. In preparation of the space experiment, we assessed the aging process under hypergravity, which is known to reduce the proliferation rate. As a result, the length of autogamy immaturity increased when measured by clock time, whereas it remained unchanged by fission age. It is therefore expected that autogamy immaturity in the measure of the clock time would be shortened under microgravity. Since the length of clonal life span of Paramecium is related to the length of autogamy immaturity, the result of hypergravity experiment supports the prediction that the clonal longevity of Paramecium under microgravity decreases. Effects of gravity on proliferation are discussed in terms of energetics of swimming during gravikinesis and gravitaxis of Paramecium.

  7. Voltage-gated calcium channels of Paramecium cilia.

    PubMed

    Lodh, Sukanya; Yano, Junji; Valentine, Megan S; Van Houten, Judith L

    2016-10-01

    Paramecium cells swim by beating their cilia, and make turns by transiently reversing their power stroke. Reversal is caused by Ca(2+) entering the cilium through voltage-gated Ca(2+) (CaV) channels that are found exclusively in the cilia. As ciliary Ca(2+) levels return to normal, the cell pivots and swims forward in a new direction. Thus, the activation of the CaV channels causes cells to make a turn in their swimming paths. For 45 years, the physiological characteristics of the Paramecium ciliary CaV channels have been known, but the proteins were not identified until recently, when the P. tetraurelia ciliary membrane proteome was determined. Three CaVα1 subunits that were identified among the proteins were cloned and confirmed to be expressed in the cilia. We demonstrate using RNA interference that these channels function as the ciliary CaV channels that are responsible for the reversal of ciliary beating. Furthermore, we show that Pawn (pw) mutants of Paramecium that cannot swim backward for lack of CaV channel activity do not express any of the three CaV1 channels in their ciliary membrane, until they are rescued from the mutant phenotype by expression of the wild-type PW gene. These results reinforce the correlation of the three CaV channels with backward swimming through ciliary reversal. The PwB protein, found in endoplasmic reticulum fractions, co-immunoprecipitates with the CaV1c channel and perhaps functions in trafficking. The PwA protein does not appear to have an interaction with the channel proteins but affects their appearance in the cilia. © 2016. Published by The Company of Biologists Ltd.

  8. Voltage-gated calcium channels of Paramecium cilia

    PubMed Central

    Lodh, Sukanya; Valentine, Megan S.; Van Houten, Judith L.

    2016-01-01

    ABSTRACT Paramecium cells swim by beating their cilia, and make turns by transiently reversing their power stroke. Reversal is caused by Ca2+ entering the cilium through voltage-gated Ca2+ (CaV) channels that are found exclusively in the cilia. As ciliary Ca2+ levels return to normal, the cell pivots and swims forward in a new direction. Thus, the activation of the CaV channels causes cells to make a turn in their swimming paths. For 45 years, the physiological characteristics of the Paramecium ciliary CaV channels have been known, but the proteins were not identified until recently, when the P. tetraurelia ciliary membrane proteome was determined. Three CaVα1 subunits that were identified among the proteins were cloned and confirmed to be expressed in the cilia. We demonstrate using RNA interference that these channels function as the ciliary CaV channels that are responsible for the reversal of ciliary beating. Furthermore, we show that Pawn (pw) mutants of Paramecium that cannot swim backward for lack of CaV channel activity do not express any of the three CaV1 channels in their ciliary membrane, until they are rescued from the mutant phenotype by expression of the wild-type PW gene. These results reinforce the correlation of the three CaV channels with backward swimming through ciliary reversal. The PwB protein, found in endoplasmic reticulum fractions, co-immunoprecipitates with the CaV1c channel and perhaps functions in trafficking. The PwA protein does not appear to have an interaction with the channel proteins but affects their appearance in the cilia. PMID:27707864

  9. Effects of static magnetic fields on growth of Paramecium caudatum.

    PubMed

    Elahee, Khouaildi B; Poinapen, Danny

    2006-01-01

    Little is known about the influence of magnetic fields on growth of primitive eukaryotes such as the ciliate Paramecium. The latter are known to exhibit interesting characteristics such as electrotaxis, gravitaxis, and membrane excitability not commonly encountered in higher organisms. This preliminary study reports the effects of static magnetic fields on growth of Paramecium caudatum. The microorganisms were either permanently or 24 h on-and-off exposed to North and South polarity magnetic fields of average field gradient 4.3 T/m, for a period of 96 h. The growth rate and lag phase of all exposed populations were not significantly different from control ones exposed to normal geomagnetic field (P > .05). However, a significant negative shift in t(max) (time taken for maximum growth) of 10.5%-12.2% and a significant decrease (P < .05) in population size of 10.2%-15.1% during the 96 h of experimental conditions were recorded for exposed populations compared to control. Our results suggest that magnetic fields, irrespective of polarity and exposure period reduce Paramecium growth by triggering early senescence of the population. The mechanisms underlying the small changes in population growth are unknown at this level, but various hypotheses have been suggested, including disorganization of swimming patterns resulting from (i) changes in cell membrane electric potential due to high speed movement through a gradient magnetic field and (ii) thermodynamic effect of anisotropic magnetic energies on cell membrane components affecting functioning of calcium channels. Altered swimming movements could in turn affect highly orchestrated processes such as conjugation, essential for survival of the organisms during development of adverse environmental conditions as thought to occur in the closed culture system used in this study.

  10. Reaction of Paramecium to Temperature Gradients and the Telegraph Equation

    NASA Astrophysics Data System (ADS)

    Sato, Tadashi; Fujimura, Jun

    1992-01-01

    The telegraph equation with a drift term to determine an asymmetric random walk is applied to the behavior of paramecia to accumulate at their optimal temperature in the presence of temperature gradient. From this equation, the following two points are clarified; 1, the stationary solution of the equation explains that the observed stationary distribution of the cells decreases exponentially from their optimal position, and 2, on a kind of paramecium that begins to accumulate at the optimal temperature after the treatment with riboflavin, the calculated transition time from a random distribution to the stationary distribution agrees with the observed transition time, 3 minutes.

  11. Ciliary membranes and mating substances in Paramecium caudatum.

    PubMed

    Watanabe, T

    1977-08-01

    Cilia detached from mating reactive cells of Paramecium caudatum were fractionated for the purpose of identifying the structural component bearing mating substances. Purified axoneme fractions had no mating reactivity. The membrane fraction obtained by dialyzing against a solution of Tris-EDTA (0.1 mm EDTA, 1 mM Tris-HCI, pH 7.6) and 0.6 m KCI, and then by centrifuging over 40% (w/v) sucrose was strongly reactive. No mating reactivity was detected in the soluble fractions containing axonemal and matrix proteins. The results indicate that the mating substances in active form are localized only on the ciliary membranes.

  12. A genetic dissection of the photophobic response of Paramecium tetraurelia.

    PubMed

    Hinrichsen, Robert; Peters, Christian

    2013-05-01

    Paramecium tetraurelia displayed two behavioral responses upon the initiation of a light stimulus at 7 x 10(4) lux. The cells exhibited a photophobic response in the form of behavioral avoiding reactions, followed by an increase in forward swimming velocity that was significantly higher than prior to the light stimulus activation. It was determined that an intensity of approximately 6.5 x 10(3) lux was required to initiate a moderate avoidance behavioral response. Following the avoiding response, a gradual increase in speed occurred as the intensity increased, indicating that increased swimming speeds are dependent on the light intensity. Two mutants, pawnA and Dancer, were utilized since they affect known Ca(2+)-currents of the cell. The use of pawnA cells, which lack voltage-dependent Ca(2+) channel activity, showed that the two responses to light could be genetically separated, in that the cells showed no avoiding reactions, but did increase their swimming speed. The Dancer cells, which display exaggerated Ca(2+) channel activity, exhibited similar initial avoiding responses as the wild type cells, however did not increase their swimming speed as the intensity of the light was increased. This phenotype as replicated in wildtype cells that had been placed in 25 μM 8-Br-cGMP. These data demonstrate that the photophobic light response of Paramecium tetraurelia can be genetically dissected as a means of elucidating the molecular mechanisms of the light response. Copyright © 2013 Elsevier GmbH. All rights reserved.

  13. Extraordinary genome stability in the ciliate Paramecium tetraurelia.

    PubMed

    Sung, Way; Tucker, Abraham E; Doak, Thomas G; Choi, Eunjin; Thomas, W Kelley; Lynch, Michael

    2012-11-20

    Mutation plays a central role in all evolutionary processes and is also the basis of genetic disorders. Established base-substitution mutation rates in eukaryotes range between ∼5 × 10(-10) and 5 × 10(-8) per site per generation, but here we report a genome-wide estimate for Paramecium tetraurelia that is more than an order of magnitude lower than any previous eukaryotic estimate. Nevertheless, when the mutation rate per cell division is extrapolated to the length of the sexual cycle for this protist, the measure obtained is comparable to that for multicellular species with similar genome sizes. Because Paramecium has a transcriptionally silent germ-line nucleus, these results are consistent with the hypothesis that natural selection operates on the cumulative germ-line replication fidelity per episode of somatic gene expression, with the germ-line mutation rate per cell division evolving downward to the lower barrier imposed by random genetic drift. We observe ciliate-specific modifications of widely conserved amino acid sites in DNA polymerases as one potential explanation for unusually high levels of replication fidelity.

  14. A protein called immaturin controlling sexual immaturity in Paramecium.

    PubMed

    Haga, N; Hiwatashi, K

    1981-01-15

    As in many metazoans, clones of some species of Paramecium have, after conjugation, a period of immaturity during which the cells cannot mate. The duration of immaturity is measured by the number of cell generations, which remains fairly constant, although duration in time varies with rate of cell reproduction. Genic involvement is shown by mutants with reduced periods of immaturity. In three different groups of Paramecium species, the cytoplasm of immature cells apparently contains the same substance which represses mating activity when injected into sexually mature cells. The immaturity-inducing substance seems to be absent from sexually mature cells, as brei made not only from mature cells in the stationary phase (mating-reactive cells), but also from those in the log phase (mating-non-reactive cells), does not repress mating activity when injected into mature cells. Variations in the amount of the substance during immaturity suggest that it controls the duration of the period. We have isolated and partially characterized the substance-a single protein called immaturin. The activity of immaturin is dose dependent and associated with a heat-labile protein of molecular weight (MW) 10,000.

  15. Extraordinary genome stability in the ciliate Paramecium tetraurelia

    PubMed Central

    Sung, Way; Tucker, Abraham E.; Doak, Thomas G.; Choi, Eunjin; Thomas, W. Kelley; Lynch, Michael

    2012-01-01

    Mutation plays a central role in all evolutionary processes and is also the basis of genetic disorders. Established base-substitution mutation rates in eukaryotes range between ∼5 × 10−10 and 5 × 10−8 per site per generation, but here we report a genome-wide estimate for Paramecium tetraurelia that is more than an order of magnitude lower than any previous eukaryotic estimate. Nevertheless, when the mutation rate per cell division is extrapolated to the length of the sexual cycle for this protist, the measure obtained is comparable to that for multicellular species with similar genome sizes. Because Paramecium has a transcriptionally silent germ-line nucleus, these results are consistent with the hypothesis that natural selection operates on the cumulative germ-line replication fidelity per episode of somatic gene expression, with the germ-line mutation rate per cell division evolving downward to the lower barrier imposed by random genetic drift. We observe ciliate-specific modifications of widely conserved amino acid sites in DNA polymerases as one potential explanation for unusually high levels of replication fidelity. PMID:23129619

  16. Improved methods and resources for paramecium genomics: transcription units, gene annotation and gene expression.

    PubMed

    Arnaiz, Olivier; Van Dijk, Erwin; Bétermier, Mireille; Lhuillier-Akakpo, Maoussi; de Vanssay, Augustin; Duharcourt, Sandra; Sallet, Erika; Gouzy, Jérôme; Sperling, Linda

    2017-06-26

    The 15 sibling species of the Paramecium aurelia cryptic species complex emerged after a whole genome duplication that occurred tens of millions of years ago. Given extensive knowledge of the genetics and epigenetics of Paramecium acquired over the last century, this species complex offers a uniquely powerful system to investigate the consequences of whole genome duplication in a unicellular eukaryote as well as the genetic and epigenetic mechanisms that drive speciation. High quality Paramecium gene models are important for research using this system. The major aim of the work reported here was to build an improved gene annotation pipeline for the Paramecium lineage. We generated oriented RNA-Seq transcriptome data across the sexual process of autogamy for the model species Paramecium tetraurelia. We determined, for the first time in a ciliate, candidate P. tetraurelia transcription start sites using an adapted Cap-Seq protocol. We developed TrUC, multi-threaded Perl software that in conjunction with TopHat mapping of RNA-Seq data to a reference genome, predicts transcription units for the annotation pipeline. We used EuGene software to combine annotation evidence. The high quality gene structural annotations obtained for P. tetraurelia were used as evidence to improve published annotations for 3 other Paramecium species. The RNA-Seq data were also used for differential gene expression analysis, providing a gene expression atlas that is more sensitive than the previously established microarray resource. We have developed a gene annotation pipeline tailored for the compact genomes and tiny introns of Paramecium species. A novel component of this pipeline, TrUC, predicts transcription units using Cap-Seq and oriented RNA-Seq data. TrUC could prove useful beyond Paramecium, especially in the case of high gene density. Accurate predictions of 3' and 5' UTR will be particularly valuable for studies of gene expression (e.g. nucleosome positioning, identification of cis

  17. Longevity of Paramecium Cell Clone under Microgravity in Space: Hypergravity Experiment as a Ground Simulation

    NASA Astrophysics Data System (ADS)

    Kato, Y.; Mogami, Y.; Baba, S. A.

    We proposed a space experiment aboard International Space Station to explore the effects of the stay under microgravity on the longevity of Paramecium cell clone (Mogami et al., 1999, Adv. Space Res., 23/12, 2087-2090). Former space experiments in CYTOS and Space Lab D-1 demonstrated that Paramecium proliferated faster in space. In combination with the fact that aging process in Paramecium is largely related to the fission age, the results of the proliferation experiment in space may predict that the longevity of Paramecium decreases when measured by clock time. As a ground simulation of the space experiment, we made an experiment to assess the aging process under hypergravity, which is known to reduce the proliferation rate. As a result, the length of autogamy immaturity increased when measured by clock time, whereas it remained unchanged by fission age (Kato et al., 2003, Zool. Sci., 1373-1380). It is therefore expected that autogamy immaturity in the measure of the clock time would be shortened under microgravity. Since the length of clonal life span of Paramecium is related to the length of autogamy immaturity, the result of hypergravity experiment may support the prediction above; i.e. a decrease in the clonal longevity of Paramecium under microgravity in space.

  18. Paramecium BBS genes are key to presence of channels in Cilia

    PubMed Central

    2012-01-01

    Background Changes in genes coding for ciliary proteins contribute to complex human syndromes called ciliopathies, such as Bardet-Biedl Syndrome (BBS). We used the model organism Paramecium to focus on ciliary ion channels that affect the beat form and sensory function of motile cilia and evaluate the effects of perturbing BBS proteins on these channels. Methods We used immunoprecipitations and mass spectrometry to explore whether Paramecium proteins interact as in mammalian cells. We used RNA interference (RNAi) and swimming behavior assays to examine the effects of BBS depletion on ciliary ion channels that control ciliary beating. Combining RNA interference and epitope tagging, we examined the effects of BBS depletion of BBS 7, 8 and 9 on the location of three channels and a chemoreceptor in cilia. Results We found 10 orthologs of 8 BBS genes in P. tetraurelia. BBS1, 2, 4, 5, 7, 8 and 9 co-immunoprecipitate. While RNAi reduction of BBS 7 and 9 gene products caused loss and shortening of cilia, RNAi for all BBS genes except BBS2 affected patterns of ciliary motility that are governed by ciliary ion channels. Swimming behavior assays pointed to loss of ciliary K+ channel function. Combining RNAi and epitope tagged ciliary proteins we demonstrated that a calcium activated K+ channel was no longer located in the cilia upon depletion of BBS 7, 8 or 9, consistent with the cells’ swimming behavior. The TRPP channel PKD2 was also lost from the cilia. In contrast, the ciliary voltage gated calcium channel was unaffected by BBS depletion, consistent with behavioral assays. The ciliary location of a chemoreceptor for folate was similarly unperturbed by the depletion of BBS 7, 8 or 9. Conclusions The co-immunoprecipitation of BBS 1,2,4,5,7,8, and 9 suggests a complex of BBS proteins. RNAi for BBS 7, 8 or 9 gene products causes the selective loss of K+ and PKD2 channels from the cilia while the critical voltage gated calcium channel and a peripheral receptor protein remain

  19. Sex recombination, and reproductive fitness: an experimental study using Paramecium

    SciTech Connect

    Nyberg, D.

    1982-08-01

    The effect of sex and recombination on reproductive fitness are measured using five wild stocks of Paramecium primaurelia. Among the wild stocks there were highly significant differences in growth rates. No hybrid had as low a fitness as the least fit parental stock. Recombination produced genotypes of higher fitness than those of either parent only in the cross between the two stocks of lowest fitness. The increase in variance of fitness as a result of recombination was almost exclusively attributable to the generation lines with low fitness. The fitness consequences of sexuality and mate choice were stock specific; some individuals leaving the most descendants by inbreeding, others by outcrossing. For most crosses the short-term advantage of sex, if any, accrue from the fusion of different gametes (hybrid vigor) and not from recombination. Since the homozygous genotype with the highest fitnes left the most progeny by inbreeding (no recombination), the persistence of conjugation in P. primaurelia is paradoxical. (JMT)

  20. An UPF3-based nonsense-mediated decay in Paramecium.

    PubMed

    Contreras, Julia; Begley, Victoria; Macias, Sandra; Villalobo, Eduardo

    2014-12-01

    Nonsense-mediated decay recognises mRNAs containing premature termination codons. One of its components, UPF3, is a molecular link bridging through its binding to the exon junction complex nonsense-mediated decay and splicing. In protists UPF3 has not been identified yet. We report that Paramecium tetraurelia bears an UPF3 gene and that it has a role in nonsense-mediated decay. Interestingly, the identified UPF3 has not conserved the essential amino acids required to bind the exon junction complex. Though, our data indicates that this ciliate bears genes coding for core proteins of the exon junction complex. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  1. Analysis of amino acid and codon usage in Paramecium bursaria.

    PubMed

    Dohra, Hideo; Fujishima, Masahiro; Suzuki, Haruo

    2015-10-07

    The ciliate Paramecium bursaria harbors the green-alga Chlorella symbionts. We reassembled the P. bursaria transcriptome to minimize falsely fused transcripts, and investigated amino acid and codon usage using the transcriptome data. Surface proteins preferentially use smaller amino acid residues like cysteine. Unusual synonymous codon and amino acid usage in highly expressed genes can reflect a balance between translational selection and other factors. A correlation of gene expression level with synonymous codon or amino acid usage is emphasized in genes down-regulated in symbiont-bearing cells compared to symbiont-free cells. Our results imply that the selection is associated with P. bursaria-Chlorella symbiosis. Copyright © 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  2. IMMUNOLOGICAL STUDIES OF ISOLATED PARTICULATES OF PARAMECIUM AURELIA

    PubMed Central

    Finger, Irving; Kaback, Michael; Kittner, Philip; Heller, Carol

    1960-01-01

    Mitochondria and other particulates—cilia, trichocysts, and "small granules"—have been isolated from several stocks of Paramecium aurelia, syngen 2. Antisera against these particles and against breis have been used to characterize the fractions by diffusion in gel. Evidence is presented for the relationship of particles, as demonstrated by immunologic cross-reactivity of the soluble antigens extracted from them. Although some antigens are unique for a fraction, cross-reacting antigens in two or more fractions, as determined by "spur" formation in agar, suggest a relationship between morphologically diverse particles. A procedure for studying cross-reactions in gels is described using the specific immobilization antigens as a model. The localization of these antigens within cilia, and perhaps trichocysts, has been confirmed. Other organelles, specifically mitochondria and "small granules," appear to alter their specificity spontaneously and reversibly during cell reproduction, a pattern reminiscent of the immobilization serotypes which can transform to one another during clonal growth. PMID:13699596

  3. Swimming velocity of Paramecium under the conditions of weightlessness.

    PubMed

    Hemmersbach-Krause, R; Briegleb, W; Vogel, K; Hader, D P

    1993-10-01

    During the 6 min-lasting "free-fall conditions" (4 x 10(-6) g) of the parabolic flight of a sounding rocket Paramecium aurelia cells showed an increase of 7.5 % in their mean swimming velocity. A detailed analysis revealed that the kinetic response was transient: after 3 min the velocity decreased to the speed of the former horizontal swimming at 1 g. Control experiments simulating the influence of vibration and hypergravity during launch of the rocket lead to the conclusion that the increase of the velocity during the parabolic flight was exclusively induced by the transition to 0 g. An increased velocity was also observed under the condition of simulated weightlessness on a fast-rotating clinostat microscope.

  4. Lack of telomere shortening during senescence in Paramecium.

    PubMed Central

    Gilley, D; Blackburn, E H

    1994-01-01

    Paramecium tetraurelia cells have a limited clonal life span and die after approximately 200 fissions if they do not undergo the process of autogamy or conjugation. To test the possibility that cellular senescence of this species is caused by telomere shortening, we analyzed the genomic DNA of the macronucleus during the clonal life span of P. tetraurelia. We found that telomeric DNA sequences were not shortened during the interval of decreased fission rate and cellular death, defined as senescence in these cells. However, the mean size of the macronuclear DNA was markedly decreased during the clonal life span. We present a model that expands upon previous proposals that accumulated DNA damage causes cellular senescence in P. tetraurelia. Images PMID:8127914

  5. Programmed DNA under-amplification in Paramecium primaurelia.

    PubMed

    Dubrana, K; Amar, L

    2000-11-01

    Ciliates are unicellular eukaryotic organisms that contain two types of nuclei throughout their vegetative life, transcriptionally active macronuclei governing the cell phenotype, and transcriptionally inert micronuclei. Following sexual reproduction, new macronuclear genomes regularly develop from micronuclear genomes through programmed DNA rearrangements that include DNA splicing, DNA fragmentation and DNA amplification. In the course of characterization of the micronuclear version of the 9.0 kb G gene, which encodes the G surface antigen in Paramecium primaurelia, we characterized a G gene duplicate. Compared with the G gene, the G gene duplicate displays features identifying it as a psi G pseudogene. About 1.6 kb upstream from the G gene, we characterized a new gene, the P gene. A related psi P putative pseudogene lies 1.6 kb upstream from the psi G pseudogene, showing that the duplicated region extends over > 15 kb and putatively defining it as a pseudogene region. Within macronuclear genomes, this region is highly under-amplified; its level never exceeds 20% of that of the corresponding G gene/P gene region in the 11 cell clones we tested. Under-amplification of the psi G pseudogene/psi P putative pseudogene region could be due to its distal position on macronuclear chromosomes, the use of alternative DNA fragmentation domains being frequent in Paramecium species, or to intrinsically lower amplification of a large genomic region. Therefore, the psi G pseudogene/psi P putative pseudogene region presented in this study provides a useful tool for the analysis of DNA fragmentation and/or amplification in eukaryotic genomes.

  6. Behavior of Paramecium sp. in solutions containing Sr and Pb: Do Paramecium sp. alter chemical forms of those metals?

    NASA Astrophysics Data System (ADS)

    Kozai, Naofumi; Ohnuki, Toshihiko; Koka, Masahi; Satoh, Takahiro; Kamiya, Tomihiro

    2011-10-01

    The behavior of Paramecium sp. (Paramecium bursaria) in aqueous solutions containing Sr and Pb was investigated to determine the role of protozoa in the migration of radionuclides in the environment. Precultured living cells of P. bursaria were exposed to aqueous solutions containing 0.01 or 0.05 mM Sr or Pb at pH 7 for 24 h. For comparison, pre-killed cells were treated with the metal solutions in the same way. Two-dimensional elemental mappings of cells were obtained by micro-PIXE. Aquatic species of Sr and Pb were analyzed by size exclusion chromatography (SEC) coupled online to ultraviolet (UV) spectroscopy and inductivity coupled plasma mass spectroscopy (ICP-MS). The amounts of Sr adsorbed or taken up by the cells surviving for 24 h and adsorbed on pre-killed cells were below the detection limit. Cells of P. bursaria adsorbed or took up a fraction of Pb. The Pb adsorbed or taken up by the cells surviving for 24 h in the Pb solution was barely detectable, while the Pb adsorbed on pre-killed cells was clearly mappable. These findings suggest that living cells of P. bursaria have functions that reduce adsorption or uptake of Pb on the cells. Quantitative and SEC-UV-ICP-MS analyses of the Sr and Pb in aqueous phases showed no clear evidences that living cells of P. bursaria alter the chemical form of Sr or Pb remaining in the aqueous phases after the cell-solution contact.

  7. A new laboratory cultivation of Paramecium bursaria using non-pathogenic bacteria strains.

    PubMed

    Bator, Tomasz

    2010-01-01

    In most studies dealing with the laboratory cultivation of paramecia (Paramecium bursaria), Klebsiella pneumoniae bacteria are used to inoculate the medium. However, Klebsiella pneumoniae is a typical pathogen, and its use is always associated with a risk of infection. The aim of the present research was to examine non-pathogenic bacteria strains as components of the medium for Paramecium bursaria. The paramecia were incubated on lettuce infusions bacterized with different bacteria strains: Bacillus subtilis DSM 10, Bacillus megaterium DSM 32, Escherichia coli DSM 498, Micrococcus luteus DSM 348. A strain derived from the natural habitat of Paramecium bursaria was used as the control one. Experiments were conducted under constant light and in the dark. Paramecia cells were counted under a stereomicroscope on consecutive days of incubation. The obtained results show that the most intensive growth of Paramecium bursaria occurs in the presence of Escherichia coli DSM 498. The use of this strain as a component of the medium allows one to obtain a high number of ciliates regardless of the light conditions. It can be concluded that the Paramecium bursaria cultivation procedure can be modified by using the non-pathogenic bacteria strain Escherichia coli DSM 498 instead of Klebsiella pneumoniae.

  8. Evolutionary dynamics of a conserved sequence motif in the ribosomal genes of the ciliate Paramecium.

    PubMed

    Catania, Francesco; Lynch, Michael

    2010-05-04

    In protozoa, the identification of preserved motifs by comparative genomics is often impeded by difficulties to generate reliable alignments for non-coding sequences. Moreover, the evolutionary dynamics of regulatory elements in 3' untranslated regions (both in protozoa and metazoa) remains a virtually unexplored issue. By screening Paramecium tetraurelia's 3' untranslated regions for 8-mers that were previously found to be preserved in mammalian 3' UTRs, we detect and characterize a motif that is distinctly conserved in the ribosomal genes of this ciliate. The motif appears to be conserved across Paramecium aurelia species but is absent from the ribosomal genes of four additional non-Paramecium species surveyed, including another ciliate, Tetrahymena thermophila. Motif-free ribosomal genes retain fewer paralogs in the genome and appear to be lost more rapidly relative to motif-containing genes. Features associated with the discovered preserved motif are consistent with this 8-mer playing a role in post-transcriptional regulation. Our observations 1) shed light on the evolution of a putative regulatory motif across large phylogenetic distances; 2) are expected to facilitate the understanding of the modulation of ribosomal genes expression in Paramecium; and 3) reveal a largely unexplored--and presumably not restricted to Paramecium--association between the presence/absence of a DNA motif and the evolutionary fate of its host genes.

  9. Evolutionary dynamics of a conserved sequence motif in the ribosomal genes of the ciliate Paramecium

    PubMed Central

    2010-01-01

    Background In protozoa, the identification of preserved motifs by comparative genomics is often impeded by difficulties to generate reliable alignments for non-coding sequences. Moreover, the evolutionary dynamics of regulatory elements in 3' untranslated regions (both in protozoa and metazoa) remains a virtually unexplored issue. Results By screening Paramecium tetraurelia's 3' untranslated regions for 8-mers that were previously found to be preserved in mammalian 3' UTRs, we detect and characterize a motif that is distinctly conserved in the ribosomal genes of this ciliate. The motif appears to be conserved across Paramecium aurelia species but is absent from the ribosomal genes of four additional non-Paramecium species surveyed, including another ciliate, Tetrahymena thermophila. Motif-free ribosomal genes retain fewer paralogs in the genome and appear to be lost more rapidly relative to motif-containing genes. Features associated with the discovered preserved motif are consistent with this 8-mer playing a role in post-transcriptional regulation. Conclusions Our observations 1) shed light on the evolution of a putative regulatory motif across large phylogenetic distances; 2) are expected to facilitate the understanding of the modulation of ribosomal genes expression in Paramecium; and 3) reveal a largely unexplored--and presumably not restricted to Paramecium--association between the presence/absence of a DNA motif and the evolutionary fate of its host genes. PMID:20441586

  10. Morphological and Molecular Characterization of Paramecium (Viridoparamecium nov. subgen.) chlorelligerum Kahl, 1935 (Ciliophora)

    PubMed Central

    KREUTZ, MARTIN; STOECK, THORSTEN; FOISSNER, WILHELM

    2013-01-01

    We redescribe Paramecium chlorelligerum, a forgotten species, which Kahl (1935) briefly but precisely described in the addendum to his ciliate monographs as a Paramecium with symbiotic green algae. The redescription is based on classical morphological methods and the analysis of the small subunit (SSU) rDNA. Morphologically, P. chlorelligerum differs from P. (Chloroparamecium) bursaria, the second green species in the genus, by having a special swimming shape, the length of the caudal cilia, the size of the micronucleus, the size of the symbiotic algae, the contractile vacuoles (with collecting vesicles vs. collecting canals), and the number of excretory pores /contractile vacuole (1 vs. 2--3). The molecular investigations show that P. chlorelligerum forms a distinct branch distant from the P. (Chloroparamecium) bursaria clade. Thus, we classify P. chlorelligerum in a new subgenus: Paramecium (Viridoparamecium) chlorelligerum. The symbiotic alga belongs to the little-known genus Meyerella, as yet recorded only from the plankton of a North American lake. PMID:22827482

  11. Evolutionary conservancy of the endocytic and trafficking machinery in the unicellular eukaryote Paramecium.

    PubMed

    Surmacz, Liliana; Wiejak, Jolanta; Wyroba, Elzbieta

    2003-01-01

    Molecular search for the homologues of the mammalian proteins in the unicellular eukaryote Paramecium involved in endocytosis and membrane trafficking is discussed. We cloned and sequenced the gene fragments encoding the following components participating in endosome formation, sorting and maturation of the proprotein precursors, respectively, dynamin 2, Rab7 and furin. There is a proof that all these genes are expressed in this unicellular organism. The function of the identified immunoanalogues of the above described components of Paramecium endocytic machinery as well as a high degree of sequence homology to the respective human counterparts points to the evolutionary conservancy of these pathways.

  12. Mechanics of membrane-cytoskeleton attachment in Paramecium

    NASA Astrophysics Data System (ADS)

    Campillo, C.; Jerber, J.; Fisch, C.; Simoes-Betbeder, M.; Dupuis-Williams, P.; Nassoy, P.; Sykes, C.

    2012-12-01

    In this paper we assess the role of the protein MKS1 (Meckel syndrome type 1) in the cortical membrane mechanics of the ciliated protist Paramecium. This protein is known to be crucial in the process of cilium formation, and we investigate its putative role in membrane-cytoskeleton attachment. Therefore, we compare cells where the gene coding for MKS1 is silenced to wild-type cells. We found that scanning electron microscopy observation of the cell surface reveals a cup-like structure in wild-type cells that is lost in silenced cells. Since this structure is based on the underlying cytoskeleton, one hypothesis to explain this observation is a disruption of membrane attachment to the cytoskeleton in the absence of MKS1 that should affect plasma membrane mechanics. We test this by probing the mechanics of wild-type and silenced cells by micropipette aspiration. Strikingly, we observe that, at the same aspiration pressure, the membrane of silenced cells is easily aspirated by the micropipette whereas that of wild-type cells enters only at a moderate velocity, an effect that suggests a detachment of the membrane from the underlying cytoskeleton in silenced cells. We quantify this detachment by measuring the deformation of the cell cortex and the rate of cell membrane entry in the micropipette. This study offers a new perspective for the characterization of membrane-cytoskeleton attachment in protists and paves the way for a better understanding of the role of membrane-cortex attachment in cilium formation.

  13. Proliferation kinetics of paramecium tetraurelia in balloon-borne experiments

    SciTech Connect

    Croute, F.; Soleilhavoup, J.P.; Vidal, S.; Rousseille, R.; Planel, H.

    1982-06-01

    Experiments were carried out to demonstrate the effect of cosmic radiation, at a balloon-flight ceiling of about 36,500 m (120,000 ft) on single-cell organism proliferation. Paramecium tetraurelia were placed in air-tight containers and maintained at 25 degrees +/- 0.1 degrees C. Cellular growth was determined by cell count, either after recovery or during the flight, by means of an automatic fixation device. Dosimetry was performed by a tissue equivalent proportional counter and was of about 0.5 mrad/h. Flight ceiling duration ranged from 48 min - 22 h. A secondary stimulating effect of growth rate, preceded by a temporary decrease, was observed after recovery. Because of the high bacterial concentration in the trans-Mediterranean flight culture medium, the temporary drop of the growth rate, due to the radiolysis products, disappears. Researchers consider that the stimulating effect can be the result of enzymatic intracellular scavenging of radiolysis products generated in the cell.

  14. Ciliate Paramecium is a natural reservoir of Legionella pneumophila.

    PubMed

    Watanabe, Kenta; Nakao, Ryo; Fujishima, Masahiro; Tachibana, Masato; Shimizu, Takashi; Watarai, Masahisa

    2016-04-15

    Legionella pneumophila, the causative agent of Legionnaires' disease, replicates within alveolar macrophages and free-living amoebae. However, the lifestyle of L. pneumophila in the environment remains largely unknown. Here we established a novel natural host model of L. pneumophila endosymbiosis using the ciliate Paramecium caudatum. We also identified Legionella endosymbiosis-modulating factor A (LefA), which contributes to the change in life stage from endosymbiosis to host lysis, enabling escape to the environment. We isolated L. pneumophila strains from the environment, and they exhibited cytotoxicity toward P. caudatum and induced host lysis. Acidification of the Legionella-containing vacuole (LCV) was inhibited, and enlarged LCVs including numerous bacteria were observed in P. caudatum infected with L. pneumophila. An isogenic L. pneumophila lefA mutant exhibited decreased cytotoxicity toward P. caudatum and impaired the modification of LCVs, resulting in the establishment of endosymbiosis between them. Our results suggest that L. pneumophila may have a mechanism to switch their endosymbiosis in protistan hosts in the environment.

  15. Ciliate Paramecium is a natural reservoir of Legionella pneumophila

    NASA Astrophysics Data System (ADS)

    Watanabe, Kenta; Nakao, Ryo; Fujishima, Masahiro; Tachibana, Masato; Shimizu, Takashi; Watarai, Masahisa

    2016-04-01

    Legionella pneumophila, the causative agent of Legionnaires’ disease, replicates within alveolar macrophages and free-living amoebae. However, the lifestyle of L. pneumophila in the environment remains largely unknown. Here we established a novel natural host model of L. pneumophila endosymbiosis using the ciliate Paramecium caudatum. We also identified Legionella endosymbiosis-modulating factor A (LefA), which contributes to the change in life stage from endosymbiosis to host lysis, enabling escape to the environment. We isolated L. pneumophila strains from the environment, and they exhibited cytotoxicity toward P. caudatum and induced host lysis. Acidification of the Legionella-containing vacuole (LCV) was inhibited, and enlarged LCVs including numerous bacteria were observed in P. caudatum infected with L. pneumophila. An isogenic L. pneumophila lefA mutant exhibited decreased cytotoxicity toward P. caudatum and impaired the modification of LCVs, resulting in the establishment of endosymbiosis between them. Our results suggest that L. pneumophila may have a mechanism to switch their endosymbiosis in protistan hosts in the environment.

  16. Developmentally programmed excision of internal DNA sequences in Paramecium aurelia.

    PubMed

    Gratias, A; Bétermier, M

    2001-01-01

    The development of a new somatic nucleus (macronucleus) during sexual reproduction of the ciliate Paramecium aurelia involves reproducible chromosomal rearrangements that affect the entire germline genome. Macronuclear development can be induced experimentally, which makes P. aurelia an attractive model for the study of the mechanism and the regulation of DNA rearrangements. Two major types of rearrangements have been identified: the fragmentation of the germline chromosomes, followed by the formation of the new macronuclear chromosome ends in association with imprecise DNA elimination, and the precise excision of internal eliminated sequences (IESs). All IESs identified so far are short, A/T rich and non-coding elements. They are flanked by a direct repeat of a 5'-TA-3' dinucleotide, a single copy of which remains at the macronuclear junction after excision. The number of these single-copy sequences has been estimated to be around 60,000 per haploid genome. This review focuses on the current knowledge about the genetic and epigenetic determinants of IES elimination in P. aurelia, the analysis of excision products, and the tightly regulated timing of excision throughout macronuclear development. Several models for the molecular mechanism of IES excision will be discussed in relation to those proposed for DNA elimination in other ciliates.

  17. Ciliate Paramecium is a natural reservoir of Legionella pneumophila

    PubMed Central

    Watanabe, Kenta; Nakao, Ryo; Fujishima, Masahiro; Tachibana, Masato; Shimizu, Takashi; Watarai, Masahisa

    2016-01-01

    Legionella pneumophila, the causative agent of Legionnaires’ disease, replicates within alveolar macrophages and free-living amoebae. However, the lifestyle of L. pneumophila in the environment remains largely unknown. Here we established a novel natural host model of L. pneumophila endosymbiosis using the ciliate Paramecium caudatum. We also identified Legionella endosymbiosis-modulating factor A (LefA), which contributes to the change in life stage from endosymbiosis to host lysis, enabling escape to the environment. We isolated L. pneumophila strains from the environment, and they exhibited cytotoxicity toward P. caudatum and induced host lysis. Acidification of the Legionella-containing vacuole (LCV) was inhibited, and enlarged LCVs including numerous bacteria were observed in P. caudatum infected with L. pneumophila. An isogenic L. pneumophila lefA mutant exhibited decreased cytotoxicity toward P. caudatum and impaired the modification of LCVs, resulting in the establishment of endosymbiosis between them. Our results suggest that L. pneumophila may have a mechanism to switch their endosymbiosis in protistan hosts in the environment. PMID:27079173

  18. Ionic conductances of membranes in ciliated and deciliated Paramecium.

    PubMed Central

    Machemer, H; Ogura, A

    1979-01-01

    1. Paramecium caudatum was deciliated with ethanol. The ionic conductance of the membrane was investigated with constant current, voltage clamp and mechanical stimuli. 2. The resting potential was not modified by the removal of the cilia. The dependence of the resting potential on the extracellular concentrations of Ca and K was the same in deciliated and control cells. 3. The input resistance in deciliated and ciliated cells increased after the ethanol treatment. 4. The membrane capacitance decreased to 48% after deciliation, suggesting that the ciliary surface area is equal to the somatic surface area. 5. Deciliation completely removed the regenerative response (graded action potential) elicited by depolarizing current pulses or mechanical stimuli. 6. Deciliated cells retained the depolarizing and hyperpolarizing mechanoreceptor responses. 7. Voltage-clamp experiments demonstrated the loss of the early inward current in deciliated cells; it was restored during ciliary regeneration. Steady-state current-voltage relationships were unchanged by deciliation. 8. The time courses of the recovery of the membrane capacitance and of the early inward current were similar, suggesting that the number of voltage-sensitive Ca channels is proportional to the ciliary membrane area. 9. We conclude that the voltage-sensitive Ca channels reside in the ciliary membrane (in confirmation of Dunlap, 1976; Ogura & Takahashi, 1976), while mechanoreceptor channels, rectifier channels and resting conductances are localized in the somatic membrane. PMID:529122

  19. Thermal performance curves of Paramecium caudatum: a model selection approach.

    PubMed

    Krenek, Sascha; Berendonk, Thomas U; Petzoldt, Thomas

    2011-05-01

    The ongoing climate change has motivated numerous studies investigating the temperature response of various organisms, especially that of ectotherms. To correctly describe the thermal performance of these organisms, functions are needed which sufficiently fit to the complete optimum curve. Surprisingly, model-comparisons for the temperature-dependence of population growth rates of an important ectothermic group, the protozoa, are still missing. In this study, temperature reaction norms of natural isolates of the freshwater protist Paramecium caudatum were investigated, considering nearly the entire temperature range. These reaction norms were used to estimate thermal performance curves by applying a set of commonly used model functions. An information theory approach was used to compare models and to identify the best ones for describing these data. Our results indicate that the models which can describe negative growth at the high- and low-temperature branch of an optimum curve are preferable. This is a prerequisite for accurately calculating the critical upper and lower thermal limits. While we detected a temperature optimum of around 29 °C for all investigated clonal strains, the critical thermal limits were considerably different between individual clones. Here, the tropical clone showed the narrowest thermal tolerance, with a shift of its critical thermal limits to higher temperatures.

  20. THE MECHANISM OF THE NEPHRIDIAL APPARATUS OF PARAMECIUM MULTIMICRONUCLEATUM

    PubMed Central

    Organ, Alan E.; Bovee, Eugene C.; Jahn, Theodore L.

    1969-01-01

    Our recent analysis of the nephridial apparatus of Paramecium multimicronucleatum by high-speed cinematography (300 fps at X 250) indicates that before the water expulsion vesicle ("contractile vacuole") is completely voided of fluid during expulsion, the ampullae surrounding and confluent with the vesicle swell with fluid entering from their respective nephridial tubules. Once the membranes of the excretory pore at the base of the excretory canal (leading from the vesicle proper to the outside) have constricted and resealed the excretory pore, the up till then constricted injection tubules of the ampullae which conduct fluid to the vesicle open as waves of contraction along the coacervate gel around the ampulla and proceed along each ampulla from distal to proximal end. The coacervate gel around any one ampulla does not necessarily contract in phase with that of any other ampulla. Each ampulla acts independently. The fluid from the ampullae is thus pumped sequentially, but not in predetermined order, into the water expulsion vesicle, refilling and distending it. Our previous studies (Organ et al., 1968a) suggest that an actomyosinoid ATP-using mechanism may be functional in the ampullary contractions. PMID:5761920

  1. Simultaneous Evaluation of Life Cycle Dynamics between a Host Paramecium and the Endosymbionts of Paramecium bursaria Using Capillary Flow Cytometry

    PubMed Central

    Takahashi, Toshiyuki

    2016-01-01

    Endosymbioses are driving forces underlying cell evolution. The endosymbiosis exhibited by Paramecium bursaria is an excellent model with which to study symbiosis. A single-cell microscopic analysis of P. bursaria reveals that endosymbiont numbers double when the host is in the division phase. Consequently, endosymbionts must arrange their cell cycle schedule if the culture-condition-dependent change delays the generation time of P. bursaria. However, it remains poorly understood whether endosymbionts keep pace with the culture-condition-dependent behaviors of P. bursaria, or not. Using microscopy and flow cytometry, this study investigated the life cycle behaviors occurring between endosymbionts and the host. To establish a connection between the host cell cycle and endosymbionts comprehensively, multivariate analysis was applied. The multivariate analysis revealed important information related to regulation between the host and endosymbionts. Results show that dividing endosymbionts underwent transition smoothly from the division phase to interphase, when the host was in the logarithmic phase. In contrast, endosymbiont division stagnated when the host was in the stationary phase. This paper explains that endosymbionts fine-tune their cell cycle pace with their host and that a synchronous life cycle between the endosymbionts and the host is guaranteed in the symbiosis of P. bursaria. PMID:27531180

  2. Control of development of the oral apparatus of Paramecium during sexual reproduction: an embryological perspective.

    PubMed

    Ng, S F; Fujishima, M

    1989-08-01

    This study shows that development of the new soma during sexual reproduction in ciliates can be conceptualized on the same basis as embryogenesis in multicellular organisms. In conjugating Paramecium, development of a new oral apparatus takes place during fertilization and the first three divisions of the zygotic nucleus and completes well before the postsexual cell undergoes the first cell fission. The control of oral development is analyzed by microsurgical removal of the zygotic nucleus or the postzygotic nuclei from conjugants. The enucleated exconjugants can pass through an early hurdle in oral development (the initiation of oral membranelle assembly) and subsequently develop an oral apparatus. Such oral apparatuses nevertheless exhibit structural and functional abnormalities including fragmentation and misalignment of oral membranelles, absence of the postoral microtubular bundle, reduction in the length of buccal cavity, and impaired phagocytosis. Other stomatogenic aspects, such as the arrangement of basal bodies in the oral membranelles, remain unaffected. The two groups of exconjugants, one derived from cells enucleated at the zygotic stage, and the other at the postzygotic stage, exhibit the same types of oral abnormality. We conclude that (i) the zygotic nucleus is not essential for the initiation of oral membranelle assembly. The existence of zygotic signals for subsequent oral development is not ruled out, but these are insufficient. (ii) Postzygotic nuclei, as well as maternal nuclei (the old somatic nucleus and meiotic derivatives of the germ nucleus), control oral development. This reveals a parallelism between postsexual development in ciliates and the early embryology of multicellular organisms, in their reliance on information provided by maternal, as well as early postzygotic nuclei. (iii) The activity of the old somatic nucleus alone is not sufficient for the later stages of oral development. Probably, some stomatogenic functions of the old

  3. A calcium-dependent potassium current is increased by a single-gene mutation in Paramecium.

    PubMed

    Hennessey, T M; Kung, C

    1987-01-01

    The membrane currents of wild type Paramecium tetraurelia and the behavioral mutant teaA were analyzed under voltage clamp. The teaA mutant was shown to have a greatly increased outward current which was blocked completely by the combined use of internally delivered Cs+ and external TEA+. This, along with previous work (Satow, Y., Kung, C., 1976, J. Exp. Biol. 65:51-63) identified this as a K+ current. It was further found to be a calcium-activated K+ current since this increased outward K+ current cannot be elicited when the internal calcium is buffered with injected EGTA. The mutation pwB, which blocks the inward calcium current, also blocks this increased outward K+ current in teaA. This shows that this mutant current is activated by calcium through the normal depolarization-sensitive calcium channel. While tail current decay kinetic analysis showed that the apparent inactivation rates for this calcium-dependent K+ current are the same for mutant and wild type, the teaA current activates extremely rapidly. It is fully activated within 2 msec. This early activation of such a large outward current causes a characteristic reduction in the amplitude of the action potential of the teaA mutant. The teaA mutation had no effect on any of the other electrophysiological parameters examined. The phenotype of the teaA mutant is therefore a general decrease in responsiveness to depolarizing stimuli because of a rapidly activating calcium-dependent K+ current which prematurely repolarizes the action potential.

  4. Surface interactions affect the toxicity of engineered metal oxide nanoparticles toward Paramecium.

    PubMed

    Li, Kungang; Chen, Ying; Zhang, Wen; Pu, Zhichao; Jiang, Lin; Chen, Yongsheng

    2012-08-20

    To better understand the potential impacts of engineered metal oxide nanoparticles (NPs) in the ecosystem, we investigated the acute toxicity of seven different types of engineered metal oxide NPs against Paramecium multimicronucleatum, a ciliated protozoan, using the 48 h LC(50) (lethal concentration, 50%) test. Our results showed that the 48 h LC(50) values of these NPs to Paramecium ranged from 0.81 (Fe(2)O(3) NPs) to 9269 mg/L (Al(2)O(3) NPs); their toxicity to Paramecium increased as follows: Al(2)O(3) < TiO(2) < CeO(2) < ZnO < SiO(2) < CuO < Fe(2)O(3) NPs. On the basis of the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, interfacial interactions between NPs and cell membrane were evaluated, and the magnitude of interaction energy barrier correlated well with the 48 h LC(50) data of NPs to Paramecium; this implies that metal oxide NPs with strong association with the cell surface might induce more severe cytotoxicity in unicellular organisms.

  5. Pdsg1 and Pdsg2, Novel Proteins Involved in Developmental Genome Remodelling in Paramecium

    PubMed Central

    Hoehener, Cristina; Singh, Aditi; Swart, Estienne C.; Nowacki, Mariusz

    2014-01-01

    The epigenetic influence of maternal cells on the development of their progeny has long been studied in various eukaryotes. Multicellular organisms usually provide their zygotes not only with nutrients but also with functional elements required for proper development, such as coding and non-coding RNAs. These maternally deposited RNAs exhibit a variety of functions, from regulating gene expression to assuring genome integrity. In ciliates, such as Paramecium these RNAs participate in the programming of large-scale genome reorganization during development, distinguishing germline-limited DNA, which is excised, from somatic-destined DNA. Only a handful of proteins playing roles in this process have been identified so far, including typical RNAi-derived factors such as Dicer-like and Piwi proteins. Here we report and characterize two novel proteins, Pdsg1 and Pdsg2 (Paramecium protein involved in Development of the Somatic Genome 1 and 2), involved in Paramecium genome reorganization. We show that these proteins are necessary for the excision of germline-limited DNA during development and the survival of sexual progeny. Knockdown of PDSG1 and PDSG2 genes affects the populations of small RNAs known to be involved in the programming of DNA elimination (scanRNAs and iesRNAs) and chromatin modification patterns during development. Our results suggest an association between RNA-mediated trans-generational epigenetic signal and chromatin modifications in the process of Paramecium genome reorganization. PMID:25397898

  6. Molecular identification of 26 syntaxin genes and their assignment to the different trafficking pathways in Paramecium.

    PubMed

    Kissmehl, Roland; Schilde, Christina; Wassmer, Thomas; Danzer, Carsten; Nuehse, Kathrin; Lutter, Kaya; Plattner, Helmut

    2007-05-01

    SNARE proteins have been classified as vesicular (v)- and target (t)-SNAREs and play a central role in the various membrane interactions in eukaryotic cells. Based on the Paramecium genome project, we have identified a multigene family of at least 26 members encoding the t-SNARE syntaxin (PtSyx) that can be grouped into 15 subfamilies. Paramecium syntaxins match the classical build-up of syntaxins, being 'tail-anchored' membrane proteins with an N-terminal cytoplasmic domain and a membrane-bound single C-terminal hydrophobic domain. The membrane anchor is preceded by a conserved SNARE domain of approximately 60 amino acids that is supposed to participate in SNARE complex assembly. In a phylogenetic analysis, most of the Paramecium syntaxin genes were found to cluster in groups together with those from other organisms in a pathway-specific manner, allowing an assignment to different compartments in a homology-dependent way. However, some of them seem to have no counterparts in metazoans. In another approach, we fused one representative member of each of the syntaxin isoforms to green fluorescent protein and assessed the in vivo localization, which was further supported by immunolocalization of some syntaxins. This allowed us to assign syntaxins to all important trafficking pathways in Paramecium.

  7. Pdsg1 and Pdsg2, novel proteins involved in developmental genome remodelling in Paramecium.

    PubMed

    Arambasic, Miroslav; Sandoval, Pamela Y; Hoehener, Cristina; Singh, Aditi; Swart, Estienne C; Nowacki, Mariusz

    2014-01-01

    The epigenetic influence of maternal cells on the development of their progeny has long been studied in various eukaryotes. Multicellular organisms usually provide their zygotes not only with nutrients but also with functional elements required for proper development, such as coding and non-coding RNAs. These maternally deposited RNAs exhibit a variety of functions, from regulating gene expression to assuring genome integrity. In ciliates, such as Paramecium these RNAs participate in the programming of large-scale genome reorganization during development, distinguishing germline-limited DNA, which is excised, from somatic-destined DNA. Only a handful of proteins playing roles in this process have been identified so far, including typical RNAi-derived factors such as Dicer-like and Piwi proteins. Here we report and characterize two novel proteins, Pdsg1 and Pdsg2 (Paramecium protein involved in Development of the Somatic Genome 1 and 2), involved in Paramecium genome reorganization. We show that these proteins are necessary for the excision of germline-limited DNA during development and the survival of sexual progeny. Knockdown of PDSG1 and PDSG2 genes affects the populations of small RNAs known to be involved in the programming of DNA elimination (scanRNAs and iesRNAs) and chromatin modification patterns during development. Our results suggest an association between RNA-mediated trans-generational epigenetic signal and chromatin modifications in the process of Paramecium genome reorganization.

  8. Mating types in Paramecium and a molecular approach to their determination.

    PubMed

    Sawka, Natalia

    2012-01-01

    Mating types are expressed in ciliates for the duration of the mature period of their clonal cycle. During cell conjugation the reciprocal fertilization of complementary mating types takes place. Models of mating type determination in the Paramecium aurelia species complex based on classical genetics are reviewed including molecular aspects of the studies.

  9. INDUCTION OF CONJUGATION BY CELL-FREE PREPARATIONS IN PARAMECIUM MULTIMICRONUCLEATUM.

    PubMed

    MIYAKE, A

    1964-12-18

    A cell-free preparation composed mainly of cilia, obtained by treatment with K(2)Cr(2)O(7) from stocks of Paramecium multimicronucleatum (syngen 2) pure for mating type, can regularly induce conjugation in intact animals of the opposite mating type.

  10. Clonal Age and the Proportion of Defective Progeny after Autogamy in PARAMECIUM AURELIA

    PubMed Central

    Fukushima, Shinichi

    1975-01-01

    The relation of mortality and the proportion of progeny with reduced fission after autogamy to the clonal age in Paramecium aurelia was investigated. This relation is not linear but the proportion of defective progeny increases stepwise. The observations are in agreement with those expected from the calculations of the number of deleterious mutations in the micronucleus. PMID:1126627

  11. The Use of Paramecium to Observe the Toxic Effect of Cigarette Smoke.

    ERIC Educational Resources Information Center

    Bardell, David

    1986-01-01

    Describes a laboratory experiment in which Paramecium caudatum was used to demonstrate the toxic effect of cigarette smoke on the cilia of epithelium cells lining the trachea and bronchi of smokers. Provides background information and explains the procedure, including how to make a simple mechanical smoking device. (TW)

  12. The Use of Paramecium to Observe the Toxic Effect of Cigarette Smoke.

    ERIC Educational Resources Information Center

    Bardell, David

    1986-01-01

    Describes a laboratory experiment in which Paramecium caudatum was used to demonstrate the toxic effect of cigarette smoke on the cilia of epithelium cells lining the trachea and bronchi of smokers. Provides background information and explains the procedure, including how to make a simple mechanical smoking device. (TW)

  13. Evaluating dosage compensation as a cause of duplicate gene retention in Paramecium tetraurelia

    PubMed Central

    Hughes, Timothy; Ekman, Diana; Ardawatia, Himanshu; Elofsson, Arne; Liberles, David A

    2007-01-01

    The high retention of duplicate genes in the genome of Paramecium tetraurelia has led to the hypothesis that most of the retained genes have persisted because of constraints due to gene dosage. This and other possible mechanisms are discussed in the light of expectations from population genetics and systems biology. PMID:17521457

  14. Phagosome maturation in unicellular eukaryote Paramecium: the presence of RILP, Rab7 and LAMP-2 homologues.

    PubMed

    Wyroba, E; Surmacz, L; Osinska, M; Wiejak, J

    2007-01-01

    Phagosome maturation is a complex process enabling degradation of internalised particles. Our data obtained at the gene, protein and cellular level indicate that the set of components involved in this process and known up to now in mammalian cells is functioning in unicellular eukaryote. Rab7-interacting partners: homologues of its effector RILP (Rab-interacting lysosomal protein) and LAMP-2 (lysosomal membrane protein 2) as well as alpha7 subunit of the 26S proteasome were revealed in Paramecium phagolysosomal compartment. We identified the gene/transcript fragments encoding RILP-related proteins (RILP1 and RILP2) in Paramecium by PCR/RT-PCR and sequencing. The deduced amino acid sequences of RILP1 and RILP2 show 60.5% and 58.3% similarity, respectively, to the region involved in regulating of lysosomal morphology and dynein-dynactin recruitment of human RILP. RILP colocalised with Rab7 in Paramecium lysosomes and at phagolysosomal membrane during phagocytosis of both the latex beads and bacteria. In the same compartment LAMP-2 was present and its expression during latex internalisation was 2.5-fold higher than in the control when P2 protein fractions (100,000 x g) of equal load were quantified by immunoblotting. LAMP-2 cross-reacting polypeptide of approximately106 kDa was glycosylated as shown by fluorescent and Western analysis of the same blot preceded by PNGase F treatment. The alpha7 subunit of 26S proteasome was detected close to the phagosomal membrane in the small vesicles, in some of which it colocalised with Rab7. Immunoblotting confirmed presence of RILP-related polypeptide and a7 subunit of 26S proteasome in Paramecium protein fractions. These results suggest that Rab7, RILP and LAMP-2 may be involved in phagosome maturation in Paramecium.

  15. Variability in secondary structure of 18S ribosomal RNA as topological marker for identification of Paramecium species.

    PubMed

    Shakoori, Farah R; Tasneem, Fareeda; Al-Ghanim, K; Mahboob, S; Al-Misned, F; Jahan, Nusrat; Shakoori, Abdul Rauf

    2014-12-01

    Besides cytological and molecular applications, Paramecium is being used in water quality assessment and for determination of saprobic levels. An unambiguous identification of these unicellular eukaryotes is not only essential, but its ecological diversity must also be explored in the local environment. 18SrRNA genes of all the strains of Paramecium species isolated from waste water were amplified, cloned and sequenced. Phylogenetic comparison of the nucleotide sequences of these strains with 23 closely related Paramecium species from GenBank Database enabled identification of Paramecium multimicronucleatum and Paramecium jenningsi. Some isolates did not show significant close association with other Paramecium species, and because of their unique position in the phylogenetic tree, they were considered new to the field. In the present report, these isolates are being designated as Paramecium caudatum pakistanicus. In this article, secondary structure of 18SrRNA has also been analyzed as an additional and perhaps more reliable topological marker for species discrimination and for determining possible phylogenetic relationship between the ciliate species. On the basis of comparison of secondary structure of 18SrRNA of various isolated Paramacium strains, and among Paramecium caudatum pakistanicus, Tetrahymena thermophila, Drosophila melanogaster, and Homo sapiens, it can be deduced that variable regions are more helpful in differentiating the species at interspecific level rather than at intraspecific level. It was concluded that V3 was the least variable region in all the organisms, V2 and V7 were the longest expansion segments of D. melanogaster and there was continuous mutational bias towards G.C base pairing in H. sapiens. © 2014 Wiley Periodicals, Inc.

  16. Identification of Paramecium bursaria syngens through molecular markers--comparative analysis of three loci in the nuclear and mitochondrial DNA.

    PubMed

    Greczek-Stachura, Magdalena; Potekhin, Alexey; Przyboś, Ewa; Rautian, Maria; Skoblo, Inna; Tarcz, Sebastian

    2012-09-01

    This is the first attempt to resolve the phylogenetic relationship between different syngens of Paramecium bursaria and to investigate at a molecular level the intraspecific differentiation of strains originating from very distant geographical locations. Herein we introduce a new collection of five P. bursaria syngens maintained at St Petersburg State University, as the international collection of syngens was lost in the 1960s. To analyze the degree of speciation within Paramecium bursaria, we examined 26 strains belonging to five different syngens from distant and geographically isolated localities using rDNA (ITS1-5.8S-ITS2-5'LSU) fragments, mitochondrial cytochrome c oxidase subunit I (COI), and H4 gene fragments. It was shown that P. bursaria strains of the same syngens cluster together in all three inferred molecular phylogenies. The genetic diversity among the studied P. bursaria strains based on rDNA sequences was rather low. The COI divergence of Paramecium bursaria was also definitely lower than that observed in the Paramecium aurelia complex. The nucleotide sequences of the H4 gene analyzed in the present study indicate the extent of genetic differences between the syngens of Paramecium bursaria. Our study demonstrates the diagnostic value of molecular markers, which are important tools in the identification of Paramecium bursaria syngens. Copyright © 2011 Elsevier GmbH. All rights reserved.

  17. Genetic differentiation of the mitochondrial cytochrome oxidase C subunit I gene in genus Paramecium (Protista, Ciliophora).

    PubMed

    Zhao, Yan; Gentekaki, Eleni; Yi, Zhenzhen; Lin, Xiaofeng

    2013-01-01

    The mitochondrial cytochrome c oxidase subunit I (COI) gene is being used increasingly for evaluating inter- and intra-specific genetic diversity of ciliated protists. However, very few studies focus on assessing genetic divergence of the COI gene within individuals and how its presence might affect species identification and population structure analyses. We evaluated the genetic variation of the COI gene in five Paramecium species for a total of 147 clones derived from 21 individuals and 7 populations. We identified a total of 90 haplotypes with several individuals carrying more than one haplotype. Parsimony network and phylogenetic tree analyses revealed that intra-individual diversity had no effect in species identification and only a minor effect on population structure. Our results suggest that the COI gene is a suitable marker for resolving inter- and intra-specific relationships of Paramecium spp.

  18. Genetic Differentiation of the Mitochondrial Cytochrome Oxidase c Subunit I Gene in Genus Paramecium (Protista, Ciliophora)

    PubMed Central

    Zhao, Yan; Gentekaki, Eleni; Yi, Zhenzhen; Lin, Xiaofeng

    2013-01-01

    Background The mitochondrial cytochrome c oxidase subunit I (COI) gene is being used increasingly for evaluating inter- and intra-specific genetic diversity of ciliated protists. However, very few studies focus on assessing genetic divergence of the COI gene within individuals and how its presence might affect species identification and population structure analyses. Methodology/Principal findings We evaluated the genetic variation of the COI gene in five Paramecium species for a total of 147 clones derived from 21 individuals and 7 populations. We identified a total of 90 haplotypes with several individuals carrying more than one haplotype. Parsimony network and phylogenetic tree analyses revealed that intra-individual diversity had no effect in species identification and only a minor effect on population structure. Conclusions Our results suggest that the COI gene is a suitable marker for resolving inter- and intra-specific relationships of Paramecium spp. PMID:24204730

  19. Using Magnetic Forces to Probe the Gravi-response of Swimming Paramecium

    NASA Astrophysics Data System (ADS)

    Guevorkian, Karine; Valles, James M., Jr.

    2004-03-01

    Paramecium Caudatum, a single celled ciliate, alters its swimming behavior when subjected to different gravity environments (e.g. centrifugation and micro-gravity). To dissect the mechanisms behind this gravi-response and that of other biological systems, we are developing the use of magnetic body forces as a means of creating a rapidly tunable, simulated variable gravity environment. Since biological materials are weakly diamagnetic, we must subject them to intense inhomogeneous magnetic fields with characteristic field-field gradient products on the order of 16 T^2/cm. We will describe experiments on Paramecium Caudatum in which we adjust their net buoyancy with magnetic forces and measure the resulting changes in their swimming behavior.

  20. Paramecium species ingest and kill the cells of the human pathogenic fungus Cryptococcus neoformans.

    PubMed

    Frager, Shalom Z; Chrisman, Cara J; Shakked, Rachel; Casadevall, Arturo

    2010-08-01

    A fundamental question in the field of medical mycology is the origin of virulence in those fungal pathogens acquired directly from the environment. In recent years, it was proposed that the virulence of certain environmental animal-pathogenic microbes, such as Cryptococcus neoformans, originated from selection pressures caused by species-specific predation. In this study, we analyzed the interaction of C. neoformans with three Paramecium spp., all of which are ciliated mobile protists. In contrast to the interaction with amoebae, some Paramecium spp. rapidly ingested C. neoformans and killed the fungus. This study establishes yet another type of protist-fungal interaction supporting the notion that animal-pathogenic fungi in the environment are under constant selection by predation.

  1. Factors Determining the Frequency of the Killer Trait within Populations of the Paramecium aurelia Complex

    PubMed Central

    Landis, Wayne G.

    1987-01-01

    The factors maintaining the cytoplasmically inherited killer trait in populations of Paramecium tetraurelia and Paramecium biaurelia were examined using, in part, computer simulation. Frequency of the K and k alleles, infection and loss of the endosymbionts, recombination during conjugation and autogamy, cytoplasmic exchange and natural selection were incorporated in a model. Infection during cytoplasmic exchange at conjugation and natural selection were factors that would increase the proportion of killers in a population. Conversely, k alleles reduced the proportion of killers in a population, acting through conjugation and autogamy. Field studies indicate that the odd mating type is prevalent in P. tetraurelia isolated from nature. Conjugation and therefore transmission by cytoplasmic transfer would be rare. Competition studies indicate a strong selective disadvantage for sensitives at concentrations found in nature. Natural selection must therefore be the factor maintaining the killer trait in P. tetraurelia. PMID:3557112

  2. ParTIES: a toolbox for Paramecium interspersed DNA elimination studies.

    PubMed

    Denby Wilkes, Cyril; Arnaiz, Olivier; Sperling, Linda

    2016-02-15

    Developmental DNA elimination occurs in a wide variety of multicellular organisms, but ciliates are the only single-celled eukaryotes in which this phenomenon has been reported. Despite considerable interest in ciliates as models for DNA elimination, no standard methods for identification and characterization of the eliminated sequences are currently available. We present the Paramecium Toolbox for Interspersed DNA Elimination Studies (ParTIES), designed for Paramecium species, that (i) identifies eliminated sequences, (ii) measures their presence in a sequencing sample and (iii) detects rare elimination polymorphisms. ParTIES is multi-threaded Perl software available at https://github.com/oarnaiz/ParTIES. ParTIES is distributed under the GNU General Public Licence v3. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  3. Clonal lifespans cultured in chemically defined medium and conventional bacterized medium in Paramecium octoaurelia.

    PubMed

    Fukushima, S; Ogawa, H; Nishikawa, T; Sasagawa, S

    1990-06-01

    The unicellular ciliate, paramecium, reproduces by binary fission, but can not continue to divide unlimitedly without sexual reproduction. We examined the clonal life span of Paramecium octaurelia stock 299 cultured in conventional bacterized medium (BM) and a chemically defined medium (DM). The cells that lived in BM divided 300 times. Although the cells in DM divided more slowly, some cells continued to divide more than 100 times. The mean life span of 90 cell lines cultured in BM was 151 +/- 49 fissions and that of 84 cell lines in DM was 68 +/- 28. When some older cells, which had been cultured in DM, were transferred to BM, most of them showed much longer life spans than those remaining in DM. The results showed that the life spans of cell clones were affected by the culture conditions.

  4. Optical Manipulation of Symbiotic Chlorella in Paramecium Bursaria Using a Fiber Axicon Microlens

    NASA Astrophysics Data System (ADS)

    Taguchi, K.; Hirota, S.; Nakayama, H.; Kunugihara, D.; Mihara, Y.

    2012-03-01

    In this paper, chemically etched axicon fiber was proposed for laser trapping of symbiotic chlorella from paramecium bursaria. We fabricated axicon micro lenses on a single-mode bare optical fiber by selective chemical etching technique. The laser beam from fiber axicon microlens was strongly focused and optical forces were sufficient to move a symbiotic chlorella. From experimental results, it was found that our proposed fiber axicon microlens was a promising tool for cell trapping without physical contact.

  5. Effects of autogamy in Paramecium tetraurelia on catalase activity and on radiosensitivity to natural ionizing radiations

    SciTech Connect

    Croute, F.; Dupouy, D.; Charley, J.P.; Soleilhavoup, J.P.; Planel, H.

    1980-02-01

    Catalase activity of Paramecium tetraurelia decreased during autogamy and recovered to normal 5 days later. Autogamy also caused changes in the ciliate's sensitivity sensitivity to natural ionizing radiations - the decrease in cell growth rate previously described in shielded cultures did not occur when autogamous cells were used. Maximum effect of shielding was observed in 11-day-old postautogamous cells. The role of the catalase in the mechanism of natural irradiation effect is discussed.

  6. Nuclear differentiation in Paramecium tetraurelia. Transplantation of vegetative micronuclei into early exconjugants.

    PubMed

    Mikami, K; Ng, S F

    1983-03-01

    The micronucleus of Paramecium tetraurelia normally gives rise to the macronucleus by going through the series of nuclear events of meiosis, mitosis and fusion during sexual reproduction. By transplanting the micronucleus from vegetative cells into early exconjugants, we have persuaded the micronucleus to take a short cut to differentiate directly into a macronucleus. This demonstrates that the course of development of the micronucleus is flexible and can be altered by stage-specific cytoplasmic factors.

  7. Identification of an algal carbon fixation-enhancing factor extracted from Paramecium bursaria.

    PubMed

    Kato, Yutaka; Imamura, Nobutaka

    2011-01-01

    The green ciliate Paramecium bursaria contains several hundred symbiotic Chlorella species. We previously reported that symbiotic algal carbon fixation is enhanced by P. bursaria extracts and that the enhancing factor is a heat-stable, low-molecular-weight, water-soluble compound. To identify the factor, further experiments were carried out. The enhancing activity remained even when organic compounds in the extract were completely combusted at 700 degrees C, suggesting that the factor is an inorganic substance. Measurement of the major cations, K+, Ca2+, and Mg2+, by an electrode and titration of the extract resulted in concentrations of 0.90 mM, 0.55 mM, and 0.21 mM, respectively. To evaluate the effect of these cations, a mixture of the cations at the measured concentrations was prepared, and symbiotic algal carbon fixation was measured in the solution. The results demonstrated that the fixation was enhanced to the same extent as with the P. bursaria extract, and thus this mixture of K+, Ca2+, and Mg2+ was concluded to be the carbon fixation-enhancing factor. There was no effect of the cation mixture on free-living C. vulgaris. Comparison of the cation concentrations of nonsymbiotic and symbiotic Paramecium extracts revealed that the concentrations of K+ and Mg2+ in nonsymbiotic Paramecium extracts were too low to enhance symbiotic algal carbon fixation, suggesting that symbiotic P. bursaria provide suitable cation conditions for photosynthesis to its symbiotic Chlorella.

  8. Morphological and molecular characterization of Paramecium (Viridoparamecium nov. subgen.) chlorelligerum Kahl (Ciliophora).

    PubMed

    Kreutz, Martin; Stoeck, Thorsten; Foissner, Wilhelm

    2012-01-01

    We redescribe Paramecium chlorelligerum, a forgotten species, which Kahl (Tierwelt Dtl., 1935, 30:651) briefly but precisely described in the addendum to his ciliate monographs as a Paramecium with symbiotic green algae. The redescription is based on classical morphological methods and the analysis of the small subunit (SSU) rDNA. Morphologically, P. chlorelligerum differs from P. (C.) bursaria, the second green species in the genus, by having a special swimming shape, the length of the caudal cilia, the size of the micronucleus, the size of the symbiotic algae, the contractile vacuoles (with collecting vesicles vs. collecting canals), and the number of excretory pores/contractile vacuole (1 vs. 2-3). The molecular investigations show that P. chlorelligerum forms a distinct branch distant from the P. (Chloroparamecium) bursaria clade. Thus, we classify P. chlorelligerum in a new subgenus: Paramecium (Viridoparamecium) chlorelligerum. The symbiotic alga belongs to the little-known genus Meyerella, as yet recorded only from the plankton of a North American lake. © 2012 The Author(s) Journal of Eukaryotic Microbiology © 2012 International Society of Protistologists.

  9. Jump if you can't take the heat: three escape gaits of Paramecium swimming

    NASA Astrophysics Data System (ADS)

    Baroud, Charles N.; Hamel, Amandine; Fisch, Cathy; Combettes, Laurent; Dupuys-Williams, Pascale

    2010-11-01

    Paramecium is able to swim at velocities reaching several times its body size per second, by beating its thousands of cilia in an organized fashion. Here we show that Paramecium has in fact three distinct swimming gaits to escape from an aggression in the form of localized heating, depending on the magnitude of the aggression: For a weak agression, normal swimming is sufficient and produces a steady swimming velocity through cilia beating. As the heating amplitude is increased, a higher acceleration and faster swimming are achieved through synchronized beating of the cilia, which later give way to the usual metachronal waves. The synchronized beating yields high initial accelerations but requires the cell to coast through the synchrnized recovery. Finally, escape from a life-threatening agression is achieved by a "jumping" gait which does not rely on the cilia but is achieved from the explosive release of a rod-like organelles in the direction of the hot spot. Measurements through high-speed video explain the role of these rods in defending Paramecium. They also show that the zero-Reynolds number assumption is unverified in most cases.

  10. Feeding of swimming Paramecium with fore-aft asymmetry in viscous fluid

    NASA Astrophysics Data System (ADS)

    Zhang, Peng; Jana, Saikat; Giarra, Matthew; Vlachos, Pavlos; Jung, Sunghwan

    2013-11-01

    Swimming behaviours and feeding efficiencies of Paramecium Multimicronucleatum with fore-aft asymmetric body shapes are studied experimentally and numerically. Among various possible swimming ways, ciliates typically exhibit only one preferred swimming directions in favorable conditions. Ciliates, like Paramecia, with fore-aft asymmetric shapes preferably swim towards the slender anterior while feeding fluid to the oral groove located at the center of the body. Since both feeding and swimming efficiencies are influenced by fluid motions around the body, it is important to reveal the fluid mechanics around a moving object. Experimentally, μ-PIV methods are employed to characterize the source-dipole streamline patterns and fluid motions around Paramecium. Numerical simulations by boundary element methods are also used to evaluate surface stresses and velocities, which give insights into the efficiencies of swimming and feeding depending on body asymmetry. It is concluded that a slender anterior and fat posterior increases the combined efficiency of swimming and feeding, which matches well with actual shapes of Paramecium. Discrepancies between experiments and simulations are also discussed.

  11. Genetic and epigenetic factors affecting meiosis induction in eukaryotes revealed in paramecium research.

    PubMed

    Prajer, Małgorzata

    2008-01-01

    This review presents studies of the induction of meiosis undertaken on the ciliate Paramecium, a unicellular model eukaryotic organism. Meiosis in Paramecium, preceding the process of fertilization, appears in starved cells after passing a defined number of divisions (cell generations), starting from the last fertilization. Investigations were performed on clones of cells entering autogamy, a self-fertilization process. Genetic as well as epigenetic factors, i.e. endo- and exogenous factors, affecting the induction ofmeiosis and changing the duration of the interautogamous interval (IAI), were analyzed. The results show that: (1) Meiosis induction is controlled genetically by the somatic macronucleus. However, besides the nuclear factors, the cytoplasmic protein immaturin also affects this process (Haga & Hiwatashi 1981); (2) Epigenetic factors, such as non-genetically disturbed cytoskeleton structures and changes in the cell architecture observed in doublet Paramecium cells, exert internal mechanical stress (Ingber 2003), which constitutes the endogenous impulse accelerating meiosis; (3) Mild osmotic stress, acting as an exogenous factor, can initiate the specific MAP kinases signaling pathway resulting in earlier meiosis induction, as in other unicellular eukaryotes (Seet & Pawson 2004).

  12. THE INSENSITIVITY OF PARAMECIUM TO CYANIDE AND EFFECTS OF IRON ON RESPIRATION

    PubMed Central

    Shoup, Charles S.; Boykin, James T.

    1931-01-01

    1. The effects of KCN and iron salts on oxygen consumption has been studied in the cell of Paramecium caudatum by manometric methods. 2. KCN solutions of strengths from M/200 to M/10,000 have been shown to produce no decrease in oxygen consumption, but have in most cases produced a very slight increase in the respiration rate. 3. The pH values were found to have little or no effect on these results. 4. Iron salts produce either no effect or a great diminution of oxygen consumption, in no case causing stimulation of rates of respiration. 5. Iron salts in neutral solutions do not penetrate the Paramecium cell nor do they cause so marked an effect as in an acid state. 6. The iron-content of Paramecium was found to be extremely small and not demonstrable by delicate tests. It is believed that iron is not combined in the cell in the form of a respiration-catalyst sensitive to cyanide. PMID:19872623

  13. THE FINE STRUCTURE OF CORTICAL COMPONENTS OF PARAMECIUM MULTIMICRONUCLEATUM

    PubMed Central

    Sedar, Albert W.; Porter, Keith R.

    1955-01-01

    The electron microscope was used to study the structure and three dimensional relationships of the components of the body cortex in thin sections of Paramecium multimicronucleatum. Micrographs of sections show that the cortex is covered externally by two closely apposed membranes (together ∼250 A thick) constituting the pellicle. Beneath the pellicle the surface of the animal is molded into ridges that form a polygonal ridgework with depressed centers. It is these ridges that give the surface of the organism its characteristic configuration and correspond to the outer fibrillar system of the light microscope image. The outer ends of the trichocysts with their hood-shaped caps are located in the centers of the anterior and posterior ridges of each polygon. The cilia extend singly from the depressed centers of the surface polygons. Each cilium shows two axial filaments with 9 peripheral and parallel filaments embedded in a matrix and the whole surrouned by a thin ciliary membrane. The 9 peripheral filaments are double and these are evenly spaced in a circle around the central pair. The ciliary membrane is continuous with the outer member of the pellicular membrane, whereas the plasma membrane is continuous with the inner member of the pellicular membrane. At the level of the plasma membrane the proximal end of the cilium is continuous with its tube-shaped basal body or kinetosome. The peripheral filaments of the cilium, together with the material of cortical matrix which tends to condense around them, form the sheath of the basal body. The kinetodesma connecting the ciliary kinetosomes (inner fibrillar system of the light microscopist) is composed of a number of discrete fibrils which overlap in a shingle-like fashion. Each striated kinetosomal fibril originates from a ciliary kinetosome and runs parallel to other kinetosomal fibrils arising from posterior kinetosomes of a particular meridional array. Sections at the level of the ciliary kinetosomes reveal an

  14. Identification of isoforms of the exocytosis-sensitive phosphoprotein PP63/parafusin in Paramecium tetraurelia and demonstration of phosphoglucomutase activity.

    PubMed Central

    Hauser, K; Kissmehl, R; Linder, J; Schultz, J E; Lottspeich, F; Plattner, H

    1997-01-01

    PP63 (parafusin) is a 63 kDa phosphoprotein which is very rapidly (within 80 ms) dephosphorylated (to P63) during triggered trichocyst exocytosis; this occurs selectively in exocytosis-competent Paramecium tetraurelia strains. In the present work, two cDNAs coding for PP63/parafusin have been isolated, one of which is a new isoform. These isoforms are 99.6% identical and are derived from two different genes. Similarity searches revealed 43-51% identity of the deduced amino acid sequences with known phosphoglucomutases from yeast and mammals. The sequences of two proteolytic peptides obtained from PP63/parafusin isolated from Paramecium are identical to parts of the amino acid sequence deduced from the major cDNA. The major cDNA was mutated from the macronuclear ciliate genetic code into the universal genetic code and expressed in Escherichia coli. The recombinant protein shows the same biochemical and immunological characteristics as the (P)P63/parafusin originally isolated from Paramecium. It has the same specific phosphoglucomutase activity as phosphoglucomutase from chicken muscle. We also show that recombinant P63-1 parafusin 1 is a substrate of an endogenous casein kinase from Paramecium, as is the originally isolated P63/parafusin. Polyclonal antibodies against recombinant P63-1/parafusin 1 were raised which recognized phosphoglucomutases from different sources. Thus we show that PP63/parafusin and phosphoglucomutase in Paramecium are identical. PMID:9173895

  15. Relationships of new sibling species of Paramecium jenningsi based on sequences of the histone H4 gene fragment.

    PubMed

    Maciejewska, Agnieszka

    2007-06-01

    Paramecium jenningsi Diller & Earl, 1958 belongs to the "aurelia" subgroup of the genus, together with Paramecium caudatum, Paramecium multimicronucleatum, Paramecium schewiakoffi and species of the Paramecium aurelia complex. The original assumption that the morphospecies P. jenningsi was a single genetic species was questioned because a comparison of genome analyses suggested the possibility that this morphospecies contained two sibling species. To refine understanding of relationships between the strains of P. jenningsi, a molecular phylogenetic analysis was conducted using H4 gene sequences. Some polymorphic sites were found among the compared sequences, and specific patterns of single nucleotide polymorphism (SNP) markers characterize two groups of strains of P. jenningsi. Phylogenetic trees constructed by different methods identified two clearly different groups (from Japan and mainland Asia) whatever the method used. The sequences of the H4 gene analyzed in the present study are closely related, and provide a good subject for phylogenetic analysis. The presence of two isolated groups of strains in the P. jenningsi group can reveal the evolutionary relationship between them; it confirms the presence of two sibling species among the known strains of P. jenningsi, and the close relationships between them and species of the P. aurelia complex.

  16. Hierarchical paramecium-like hollow and solid Au/Pt bimetallic nanostructures constructed using goethite as template.

    PubMed

    Liu, Wei; Repo, Eveliina; Heikkilä, Mikko; Leskelä, Markku; Sillanpää, Mika

    2010-10-01

    Novel hollow and solid paramecium-like hierarchical Au/Pt bimetallic nanostructures were constructed using goethite as template via a seed-mediated growth method. Transmission electron microscopy (TEM), xi-potential measurement, UV-vis spectroscopy, energy dispersive x-ray spectroscopy (EDS), ICP-AES measurement, x-ray powder diffraction (XRD) and x-ray photoelectron spectroscopy (XPS) were utilized to systematically characterize the bimetallic nanostructures. It is found that the core structure of the paramecium-like bimetallic nanomaterial is closely related to reducing agent. When ascorbic acid is used as reducing agent, goethite serves as in situ sacrificed template and hollow paramecium-like bimetallic structure is obtained. When NH(2)OH.HCl is used, solid nanostructure with preserved goethite core is produced. Heating the reaction solution is necessary to obtain the paramecium-like morphology with rough interconnected Pt cilia shell. The thickness of Pt cilia layer can be controlled by adjusting the molar ratio of H(2)PtCl(6) to Au nanoseeds. The overgrowth of the rough Pt cilia is proposed to be via an autocatalytic and three-dimensional heterogeneous nucleation process first through flower-like morphology. Both the hollow and solid hierarchical paramecium-like Au/Pt bimetallic nanostructures show good catalytic activities.

  17. Gamma-amino butyric acid (GABA) release in the ciliated protozoon Paramecium occurs by neuronal-like exocytosis.

    PubMed

    Ramoino, P; Milanese, M; Candiani, S; Diaspro, A; Fato, M; Usai, C; Bonanno, G

    2010-04-01

    Paramecium primaurelia expresses a significant amount of gamma-amino butyric acid (GABA). Paramecia possess both glutamate decarboxylase (GAD)-like and vesicular GABA transporter (vGAT)-like proteins, indicating the ability to synthesize GABA from glutamate and to transport GABA into vesicles. Using antibodies raised against mammalian GAD and vGAT, bands with an apparent molecular weight of about 67 kDa and 57 kDa were detected. The presence of these bands indicated a similarity between the proteins in Paramecium and in mammals. VAMP, syntaxin and SNAP, putative proteins of the release machinery that form the so-called SNARE complex, are present in Paramecium. Most VAMP, syntaxin and SNAP fluorescence is localized in spots that vary in size and density and are primarily distributed near the plasma membrane. Antibodies raised against mammal VAMP-3, sintaxin-1 or SNAP-25 revealed protein immunoblot bands having molecular weights consistent with those observed in mammals. Moreover, P. primaurelia spontaneously releases GABA into the environment, and this neurotransmitter release significantly increases after membrane depolarization. The depolarization-induced GABA release was strongly reduced not only in the absence of extracellular Ca(2+) but also by pre-incubation with bafilomycin A1 or with botulinum toxin C1 serotype. It can be concluded that GABA occurs in Paramecium, where it is probably stored in vesicles capable of fusion with the cell membrane; accordingly, GABA can be released from Paramecium by stimulus-induced, neuronal-like exocytotic mechanisms.

  18. Hierarchical paramecium-like hollow and solid Au/Pt bimetallic nanostructures constructed using goethite as template

    NASA Astrophysics Data System (ADS)

    Liu, Wei; Repo, Eveliina; Heikkilä, Mikko; Leskelä, Markku; Sillanpää, Mika

    2010-10-01

    Novel hollow and solid paramecium-like hierarchical Au/Pt bimetallic nanostructures were constructed using goethite as template via a seed-mediated growth method. Transmission electron microscopy (TEM), ξ-potential measurement, UV-vis spectroscopy, energy dispersive x-ray spectroscopy (EDS), ICP-AES measurement, x-ray powder diffraction (XRD) and x-ray photoelectron spectroscopy (XPS) were utilized to systematically characterize the bimetallic nanostructures. It is found that the core structure of the paramecium-like bimetallic nanomaterial is closely related to reducing agent. When ascorbic acid is used as reducing agent, goethite serves as in situ sacrificed template and hollow paramecium-like bimetallic structure is obtained. When NH2OH·HCl is used, solid nanostructure with preserved goethite core is produced. Heating the reaction solution is necessary to obtain the paramecium-like morphology with rough interconnected Pt cilia shell. The thickness of Pt cilia layer can be controlled by adjusting the molar ratio of H2PtCl6 to Au nanoseeds. The overgrowth of the rough Pt cilia is proposed to be via an autocatalytic and three-dimensional heterogeneous nucleation process first through flower-like morphology. Both the hollow and solid hierarchical paramecium-like Au/Pt bimetallic nanostructures show good catalytic activities.

  19. Delimiting Species Boundaries within a Paraphyletic Species Complex: Insights from Morphological, Genetic, and Molecular Data on Paramecium sonneborni (Paramecium aurelia species complex, Ciliophora, Protozoa).

    PubMed

    Przyboś, Ewa; Tarcz, Sebastian; Rautian, Maria; Sawka, Natalia

    2015-09-01

    The demarcation of boundaries between protist species is often problematic because of the absence of a uniform species definition, the abundance of cryptic diversity, and the occurrence of convergent morphology. The ciliates belonging to the Paramecium aurelia complex, consisting of 15 species, are a good model for such systematic and evolutionary studies. One member of the complex is P. sonneborni, previously known only from one stand in Texas (USA), but recently found in two new sampling sites in Cyprus (creeks running to Salt Lake and Oroklini Lake near Larnaca). The studied Paramecium sonneborni strains (from the USA and Cyprus) reveal low viability in the F1 and F2 generations of interstrain hybrids and may be an example of ongoing allopatric speciation. Despite its molecular distinctiveness, we postulate that P. sonneborni should remain in the P. aurelia complex, making it a paraphyletic taxon. Morphological studies have revealed that some features of the nuclear apparatus of P. sonneborni correspond to the P. aurelia spp. complex, while others are similar to P. jenningsi and P. schewiakoffi. The observed discordance indicates rapid splitting of the P. aurelia-P. jenningsi-P. schewiakoffi group, in which genetic, morphological, and molecular boundaries between species are not congruent. Copyright © 2015 Elsevier GmbH. All rights reserved.

  20. Pharmacological characterization of NMDA-like receptors in the single-celled organism Paramecium primaurelia.

    PubMed

    Ramoino, Paola; Candiani, Simona; Pittaluga, Anna Maria; Usai, Cesare; Gallus, Lorenzo; Ferrando, Sara; Milanese, Marco; Faimali, Marco; Bonanno, Giambattista

    2014-02-01

    Paramecium primaurelia is a unicellular eukaryote that moves in freshwater by ciliary beating and responds to environmental stimuli by altering motile behaviour. The movements of the cilia are controlled by the electrical changes of the cell membrane: when the intraciliary Ca(2+) concentration associated with plasma membrane depolarization increases, the ciliary beating reverses its direction, and consequently the swimming direction changes. The ciliary reversal duration is correlated with the amount of Ca(2+) influx. Here, we evaluated the effects due to the activation or blockade of N-methyl-d-aspartic acid (NMDA) receptors on swimming behaviour in Paramecium. Paramecia normally swim forward, drawing almost linear tracks. We observed that the simultaneous administration of NMDA and glycine induced a partial ciliary reversal (PaCR) leading to a continuous spiral-like swim. Furthermore, the duration of continuous ciliary reversal (CCR), triggered by high external KCl concentrations, was longer in NMDA+glycine-treated cells. NMDA action required the presence of Ca(2+), as the normal forward swimming was restored when the ion was omitted from the extracellular milieu. The PaCR and the enhancement of CCR duration significantly decreased when the antagonists of the glutamate site D-AP5 or CGS19755, the NMDA channel blocker MK-801 or the glycine site antagonist DCKA was added. The action of NMDA+glycine was also abolished by Zn(2+) or ifenprodil, the GluN2A and the GluN2B NMDA-containing subunit blockers, respectively. Searches of the Paramecium genome database currently available indicate that the NMDA-like receptor with ligand-binding characteristics of an NMDA receptor-like complex, purified from rat brain synaptic membranes and found in some metazoan genomes, is also present in Paramecium. These results provide evidence that functional NMDA receptors similar to those typical of mammalian neuronal cells are present in the single-celled organism Paramecium and thus

  1. Calcium-Release Channels in Paramecium. Genomic Expansion, Differential Positioning and Partial Transcriptional Elimination

    PubMed Central

    Ladenburger, Eva-Maria; Plattner, Helmut

    2011-01-01

    The release of Ca2+ from internal stores is a major source of signal Ca2+ in almost all cell types. The internal Ca2+ pools are activated via two main families of intracellular Ca2+-release channels, the ryanodine and the inositol 1,4,5-trisphosphate (InsP3) receptors. Among multicellular organisms these channel types are ubiquitous, whereas in most unicellular eukaryotes the identification of orthologs is impaired probably due to evolutionary sequence divergence. However, the ciliated protozoan Paramecium allowed us to prognosticate six groups, with a total of 34 genes, encoding proteins with characteristics typical of InsP3 and ryanodine receptors by BLAST search of the Paramecium database. We here report that these Ca2+-release channels may display all or only some of the characteristics of canonical InsP3 and ryanodine receptors. In all cases, prediction methods indicate the presence of six trans-membrane regions in the C-terminal domains, thus corresponding to canonical InsP3 receptors, while a sequence homologous to the InsP3-binding domain is present only in some types. Only two types have been analyzed in detail previously. We now show, by using antibodies and eventually by green fluorescent protein labeling, that the members of all six groups localize to distinct organelles known to participate in vesicle trafficking and, thus, may provide Ca2+ for local membrane-membrane interactions. Whole genome duplication can explain radiation within the six groups. Comparative and evolutionary evaluation suggests derivation from a common ancestor of canonical InsP3 and ryanodine receptors. With one group we could ascertain, to our knowledge for the first time, aberrant splicing in one thoroughly analyzed Paramecium gene. This yields truncated forms and, thus, may indicate a way to pseudogene formation. No comparable analysis is available for any other, free-living or parasitic/pathogenic protozoan. PMID:22102876

  2. Micro-particle transporting system using galvanotactically stimulated apo-symbiotic cells of Paramecium bursaria.

    PubMed

    Furukawa, Shunsuke; Karaki, Chiaki; Kawano, Tomonori

    2009-01-01

    It is well known that Paramecium species including green paramecia (Paramecium bursaria) migrate towards the anode when exposed to an electric field in a medium. This type of a cellular movement is known as galvanotaxis. Our previous study revealed that an electric stimulus given to P bursaria is converted to a galvanotactic cellular movement by involvement of T-type calcium channel on the plasma membrane [Aonuma et al. (2007), Z. Naturforsch. 62c, 93-102]. This phenomenon has attracted the attention of bioengineers in the fields of biorobotics or micro-robotics in order to develop electrically controllable micromachineries. Here, we demonstrate the galvanotactic controls of the cellular migration of P bursaria in capillary tubes (diameter, 1-2 mm; length, 30-240 mm). Since the Paramecium cells take up particles of various sizes, we attempted to use the electrically stimulated cells of P bursaria as the vehicle for transportation of micro-particles in the capillary system. By using apo-symbiotic cells of P bursaria obtained after forced removal of symbiotic algae, the uptake of the particles could be maximized and visualized. Then, electrically controlled transportations of particle-filled apo-symbiotic P bursaria cells were manifested. The particles transported by electrically controlled cells (varying in size from nm to /m levels) included re-introduced green algae, fluorescence-labeled polystyrene beads, magnetic microspheres, emerald green fluorescent protein (EmGFP)-labeled cells of E. coli, Indian ink, and crystals of zeolite (hydrated aluminosilicate minerals with a micro-porous structure) and some metal oxides. Since the above demonstrations were successful, we concluded that P bursaria has a potential to be employed as one of the micro-biorobotic devices used in BioMEMS (biological micro-electro-mechanical systems).

  3. Rare Freshwater Ciliate Paramecium chlorelligerum Kahl, 1935 and Its Macronuclear Symbiotic Bacterium "Candidatus Holospora parva".

    PubMed

    Lanzoni, Olivia; Fokin, Sergei I; Lebedeva, Natalia; Migunova, Alexandra; Petroni, Giulio; Potekhin, Alexey

    2016-01-01

    Ciliated protists often form symbioses with many diverse microorganisms. In particular, symbiotic associations between ciliates and green algae, as well as between ciliates and intracellular bacteria, are rather wide-spread in nature. In this study, we describe the complex symbiotic system between a very rare ciliate, Paramecium chlorelligerum, unicellular algae inhabiting its cytoplasm, and novel bacteria colonizing the host macronucleus. Paramecium chlorelligerum, previously found only twice in Germany, was retrieved from a novel location in vicinity of St. Petersburg in Russia. Species identification was based on both classical morphological methods and analysis of the small subunit rDNA. Numerous algae occupying the cytoplasm of this ciliate were identified with ultrastructural and molecular methods as representatives of the Meyerella genus, which before was not considered among symbiotic algae. In the same locality at least fifteen other species of "green" ciliates were found, thus it is indeed a biodiversity hot-spot for such protists. A novel species of bacterial symbionts living in the macronucleus of Paramecium chlorelligerum cells was morphologically and ultrastructurally investigated in detail with the description of its life cycle and infection capabilities. The new endosymbiont was molecularly characterized following the full-cycle rRNA approach. Furthermore, phylogenetic analysis confirmed that the novel bacterium is a member of Holospora genus branching basally but sharing all characteristics of the genus except inducing connecting piece formation during the infected host nucleus division. We propose the name "Candidatus Holospora parva" for this newly described species. The described complex system raises new questions on how these microorganisms evolve and interact in symbiosis.

  4. Non-Mendelian inheritance of macronuclear mutations is gene specific in Paramecium tetraurelia.

    PubMed

    Scott, J M; Mikami, K; Leeck, C L; Forney, J D

    1994-04-01

    Paramecium tetraurelia contains two types of nuclei, a diploid germinal micronucleus and a large transcriptionally active macronucleus. The macronuclear genome is formed from the micronuclear DNA during sexual reproduction. Previous studies have shown that the processing of the A-type variable surface protein gene during formation of a new macronucleus is dependent on the presence of the A gene in the old macronucleus. It is not clear if this is a general feature that controls the formation of the Paramecium macronuclear genome or a unique feature of the A locus. Using micronuclear transplantation, we have constructed a strain that has a wild-type micronucleus but has macronuclear deletions of the A- and B-type surface protein genes. Neither the A nor the B gene is incorporated into the new macronucleus after sexual reproduction. Macronuclear transformation of this strain with the B gene rescues the B-gene deletion after formation of the next macronucleus but has not effect on the A deletion. Similarly, transformation with the A gene shows gene-specific rescue for A but not B. The effect of the old macronucleus on the processing of the new macronucleus results in a pattern of non-Mendelian inheritance of both macronuclear deletions. Progeny from the wild-type exconjugant are all wild type, and progeny from the A- B- exconjugant are mutant. The features of this A- B- non-Mendelian mutant demonstrate that the regulation of macronuclear DNA processing is gene specific, and our results open the possibility that this type of regulation affects many regions of the Paramecium genome.

  5. A novel member of the cyclin-dependent kinase family in Paramecium tetraurelia.

    PubMed

    Zhang, H; Berger, J D

    1999-01-01

    Passage through the cell cycle in eukaryotes requires the successive activation of different cyclin-dependent protein kinases. Here, we describe the identification and characterization of a novel class of cyclin-dependent protein kinase, termed Cdk2, in the ciliate Paramecium tetraurelia. It is 301 amino acids long, 7 amino acids shorter than Cdk1, the CDK that is associated with macronuclear DNA synthesis. All the catalytic domains typical of protein kinases can be located within the sequence and putative regulatory phosphorylation sites equivalent to Thr14, Tyr15, and Thr161 in human CDK1 are also conserved. The 'PSTAIRE' region characteristic of most CDKs is perfectly conserved. Cdk2 shares only 48% homology to Cdk1 at the amino acid level, suggesting that the evolutionary separation of Cdk1 and Cdk2 is ancient, and implying that they have different roles in cell cycle regulation. Like Cdk1, Cdk2 does not bind to yeast p13suc1, even though it has better conservation of p13suc1 binding sites than Cdk1 does. The Cdk2 protein level is relatively constant throughout the vegetative cell cycle. Cdk2 exhibits kinase activity towards bovine histone H1 in vitro with the maximal level late in the cell cycle, suggesting it may be involved in the regulation of cytokinesis. Our results further support the view that an analogue of the cyclin-dependent kinase cell cycle regulatory system like that of yeast and higher eukaryotic cells operates in Paramecium and that a family of cyclin-dependent kinases may control different aspects of the Paramecium cell cycle.

  6. The ciliate Paramecium shows higher motility in non-uniform chemical landscapes.

    PubMed

    Giuffre, Carl; Hinow, Peter; Vogel, Ryan; Ahmed, Tanvir; Stocker, Roman; Consi, Thomas R; Strickler, J Rudi

    2011-04-11

    We study the motility behavior of the unicellular protozoan Paramecium tetraurelia in a microfluidic device that can be prepared with a landscape of attracting or repelling chemicals. We investigate the spatial distribution of the positions of the individuals at different time points with methods from spatial statistics and Poisson random point fields. This makes quantitative the informal notion of "uniform distribution" (or lack thereof). Our device is characterized by the absence of large systematic biases due to gravitation and fluid flow. It has the potential to be applied to the study of other aquatic chemosensitive organisms as well. This may result in better diagnostic devices for environmental pollutants.

  7. The Ciliate Paramecium Shows Higher Motility in Non-Uniform Chemical Landscapes

    PubMed Central

    Giuffre, Carl; Hinow, Peter; Vogel, Ryan; Ahmed, Tanvir; Stocker, Roman; Consi, Thomas R.; Strickler, J. Rudi

    2011-01-01

    We study the motility behavior of the unicellular protozoan Paramecium tetraurelia in a microfluidic device that can be prepared with a landscape of attracting or repelling chemicals. We investigate the spatial distribution of the positions of the individuals at different time points with methods from spatial statistics and Poisson random point fields. This makes quantitative the informal notion of “uniform distribution” (or lack thereof). Our device is characterized by the absence of large systematic biases due to gravitation and fluid flow. It has the potential to be applied to the study of other aquatic chemosensitive organisms as well. This may result in better diagnostic devices for environmental pollutants. PMID:21494596

  8. Food ingestion and egestion in mating reactive populations of Paramecium primaurelia.

    PubMed

    Ramoino, P

    1992-01-01

    The study of food ingestion and egestion carried out on Paramecium primaurelia mating reactive cells shows that, after their transfer into a medium with suspended particles, the complementary mating type cells exhibit very significant differences in the food vacuole formation and egestion rate. Under the same external environmental conditions, the mating type II cells form and egest a higher number of food vacuoles when compared with mating type I cells. The higher rate of food vacuole formation shown by the mating type II cells is related to their faster growth rate.

  9. [A computer method for the evaluation of Paramecium motor activity using video records of their movement].

    PubMed

    Bingi, V N; Zarutskiĭ, A A; Kapranov, S V; Kovalev, Iu M; Miliaev, V A; Tereshchenko, N A

    2004-01-01

    A method for the evaluation of Paramecium caudatum motility was proposed as a tool for the investigation of magnetobiological as well as other physical and chemical effects. The microscopically observed movement of paramecia is recorded and processed using a special software program. The protozoan motility is determined as a function of their mean velocity in a definite time. The main advantages of the method are that it is easily modified for determining various characteristics of the motor activity of paramecia and that the video data obtained can be reused.

  10. Multiple lines of evidence shed light on the occurrence of paramecium (ciliophora, oligohymenophorea) in bromeliad tank water.

    PubMed

    Buosi, Paulo R B; Cabral, Adalgisa F; Simão, Taiz L L; Utz, Laura R P; Velho, Luiz F M

    2014-01-01

    Phytotelmata are vegetal structures that hold water from the rain, and organic matter from the forest and the soil, resulting in small, compartmentalized bodies of water, which provide an essential environment for the establishment and development of many organisms. These microenvironments generally harbor endemic species, but many organisms that are found in lakes and rivers, are also present. Here, we report, for the first time, the occurrence of the ciliate genus Paramecium in the tank of the bromeliad species Aechmaea distichantha. The identification of the Paramecium species was performed based on live observations, protargol impregnation, scanning electronic microscopy, and sequencing of the 18s rRNA. The absence of Paramecium from bromeliad tank water was highlighted in several earlier investigations, and may be due to the fact that this species is unable to make cysts. The occurrence of Paramecium multimicronucleatum in our samples may be explained by the proximity between the bromeliads and the river, a potential source of the species. Further, we also believe that the counting methodology used in our study provides a more accurate analysis of the species diversity, since we investigated all samples within a maximum period of 6 h after sampling, allowing minimum loss of specimens. © 2013 The Author(s) Journal of Eukaryotic Microbiology © 2013 International Society of Protistologists.

  11. Paramecium tetraurelia pre-screen for hazardous chemicals: a rapid detector system for health hazards. 1977-1983 report

    SciTech Connect

    Smith-Sonneborn, J.

    1983-01-01

    The purpose was to develop and validate a new eukaryotic bioassay system applicable to rapid identification of environmental toxins, mutagens, and carcinogens. The ability of Paramecium to detect potential health hazards, associated with complex environmental mixtures, was demonstrated in association with the finest coal fly ash particles and aqueous waste streams from both oil shale and coal gasification developing technologies. In Paramecium, the cytotoxicity of an agent was determined by altering the survival and/or growth rate of single cells in test agents. Genotoxicity was assayed by a two-tiered approach, utilizing both the Paramecium system and the more established Ames Salmonella assay for mutagen/carcinogen detection. An agent was considered genotoxic in Paramwecium if altered phenotypes were induced in the fertilization progeny of treated parent cells. Since others had shown a significant correlation between agents which were photodynamically active in Paramecium and carcinogenic in mammals, the photodynamic activity versus carcinogenicity of agents was reviewed. Photodynamically active compounds are defined by the immobilization of cells when exposed to both the test agent and black light; neither the agent alone nor light alone affects swimming activity.

  12. A synthetic cadmium metallothionein gene (PMCd1syn) of Paramecium species: expression, purification and characteristics of metallothionein protein.

    PubMed

    Dar, Saira; Shuja, Rukhsana N; Shakoori, Abdul Rauf

    2013-02-01

    Metallothioneins (MTs) are metal binding proteins that are rich in cysteine residues constituting 10-30 % of the total protein, and in which the thiol groups bind to the metal ions. The increasing amount of metal ions in the medium have shown increased production of MTs by different organisms such as bacteria, protozoa and mammals like humans. PMCd1 is the first gene ever discovered in Paramecium, a ciliated protozoan, that could produce this MT in response to cadmium. In this study the PMCd1syn gene has been cloned in pET41a expression vector and expressed in an Escherichia coli BL21-codonplus strain for the first time. Since the gene PMCd1 amplified from Paramecium contained 10 codons, which could act as stop codons during expression in E. coli, this gene of 612 bps was synthesized to substitute these (stop) codons for the Paramecium sp. specific amino acids. For stability of the expressed protein, glutathione-S-transferase gene was fused with PMCd1syn gene and coexpressed. The cells expressing PMCd1syn demonstrated increased accumulation of cadmium. This is the first report of cadmium MT protein expressed from Paramecium species, particularly from synthetic MT gene (PMCd1syn). This fusion protein, the molecular weight of which has been confirmed to be 53.03 kDa with MALDI analysis, is rich in cysteine residues, and has been shown for the first time in this ciliate to bind to and sequester Cd(2+)-ions.

  13. Paramecium tetraurelia. Pre-screen for hazardous chemicals: a rapid-detector system for health hazards. Progress report

    SciTech Connect

    Smith-Sonneborn, J.

    1981-01-01

    Genotoxic and cytotoxic effects of dilutions of oil shale process waters from three different retorting processes were evaluated using the Paramecium tetraurelia and the Salmonella assays. Process waters from above-ground, true in situ, and modified retorting process were analyzed. Significant mutagenicity was detected in waste water from both the above-ground and true in situ retorting processes in both the Salmonella and Paramecium bioassays. The Salmonella assay required the addition of the rat liver S9 fraction; the Paramecium bioassay was more sensitive to genotoxic effects from an above-ground retort water without the addition of the rat liver fraction. Mutagenicity of the dilutions tested from the modified retort process was detected only in the Paramecium system. The waste water from the above-ground retort process was the most toxic to both the protozoan and bacteria. Since the chemical composition of oil shale process waters can vary with: 1) the retorting process used, 2) the temperature of the retorting, 3) the composition of the shale rock, and 4) the sampling and storage procedures, the differences in biological activity observed between water can reflect differences in water contamination.

  14. An Ins(1,4,5)P3 receptor in Paramecium is associated with the osmoregulatory system.

    PubMed

    Ladenburger, Eva-Maria; Korn, Iris; Kasielke, Nicole; Wassmer, Thomas; Plattner, Helmut

    2006-09-01

    In the ciliate Paramecium, a variety of well characterized processes are regulated by Ca2+, e.g. exocytosis, endocytosis and ciliary beat. Therefore, among protozoa, Paramecium is considered a model organism for Ca2+ signaling, although the molecular identity of the channels responsible for the Ca2+ signals remains largely unknown. We have cloned - for the first time in a protozoan - the full sequence of the gene encoding a putative inositol (1,4,5)-trisphosphate (Ins(1,4,5)P3) receptor from Paramecium tetraurelia cells showing molecular characteristics of higher eukaryotic cells. The homologously expressed Ins(1,4,5)P3-binding domain binds [3H]Ins(1,4,5)P3, whereas antibodies unexpectedly localize this protein to the osmoregulatory system. The level of Ins(1,4,5)P3-receptor expression was reduced, as shown on a transcriptional level and by immuno-staining, by decreasing the concentration of extracellular Ca2+ (Paramecium cells rapidly adjust their Ca2+ level to that in the outside medium). Fluorochromes reveal spontaneous fluctuations in cytosolic Ca2+ levels along the osmoregulatory system and these signals change upon activation of caged Ins(1,4,5)P3. Considering the ongoing expulsion of substantial amounts of Ca2+ by the osmoregulatory system, we propose here that Ins(1,4,5)P3 receptors serve a new function, i.e. a latent, graded reflux of Ca2+ to fine-tune [Ca2+] homeostasis.

  15. Epigenetic regulation of serotype expression antagonizes transcriptome dynamics in Paramecium tetraurelia

    PubMed Central

    Cheaib, Miriam; Dehghani Amirabad, Azim; Nordström, Karl J. V.; Schulz, Marcel H.; Simon, Martin

    2015-01-01

    Phenotypic variation of a single genotype is achieved by alterations in gene expression patterns. Regulation of such alterations depends on their time scale, where short-time adaptations differ from permanently established gene expression patterns maintained by epigenetic mechanisms. In the ciliate Paramecium, serotypes were described for an epigenetically controlled gene expression pattern of an individual multigene family. Paradoxically, individual serotypes can be triggered in Paramecium by alternating environments but are then stabilized by epigenetic mechanisms, thus raising the question to which extend their expression follows environmental stimuli. To characterize environmental adaptation in the context of epigenetically controlled serotype expression, we used RNA-seq to characterize transcriptomes of serotype pure cultures. The resulting vegetative transcriptome resource is first analysed for genes involved in the adaptive response to the altered environment. Secondly, we identified groups of genes that do not follow the adaptive response but show co-regulation with the epigenetically controlled serotype system, suggesting that their gene expression pattern becomes manifested by similar mechanisms. In our experimental set-up, serotype expression and the entire group of co-regulated genes were stable among environmental changes and only heat-shock genes altered expression of these gene groups. The data suggest that the maintenance of these gene expression patterns in a lineage represents epigenetically controlled robustness counteracting short-time adaptation processes. PMID:26231545

  16. Insights into three whole-genome duplications gleaned from the Paramecium caudatum genome sequence.

    PubMed

    McGrath, Casey L; Gout, Jean-Francois; Doak, Thomas G; Yanagi, Akira; Lynch, Michael

    2014-08-01

    Paramecium has long been a model eukaryote. The sequence of the Paramecium tetraurelia genome reveals a history of three successive whole-genome duplications (WGDs), and the sequences of P. biaurelia and P. sexaurelia suggest that these WGDs are shared by all members of the aurelia species complex. Here, we present the genome sequence of P. caudatum, a species closely related to the P. aurelia species group. P. caudatum shares only the most ancient of the three WGDs with the aurelia complex. We found that P. caudatum maintains twice as many paralogs from this early event as the P. aurelia species, suggesting that post-WGD gene retention is influenced by subsequent WGDs and supporting the importance of selection for dosage in gene retention. The availability of P. caudatum as an outgroup allows an expanded analysis of the aurelia intermediate and recent WGD events. Both the Guanine+Cytosine (GC) content and the expression level of preduplication genes are significant predictors of duplicate retention. We find widespread asymmetrical evolution among aurelia paralogs, which is likely caused by gradual pseudogenization rather than by neofunctionalization. Finally, cases of divergent resolution of intermediate WGD duplicates between aurelia species implicate this process acts as an ongoing reinforcement mechanism of reproductive isolation long after a WGD event. Copyright © 2014 by the Genetics Society of America.

  17. A forward genetic screen reveals essential and non-essential RNAi factors in Paramecium tetraurelia

    PubMed Central

    Marker, Simone; Carradec, Quentin; Tanty, Véronique; Arnaiz, Olivier; Meyer, Eric

    2014-01-01

    In most eukaryotes, small RNA-mediated gene silencing pathways form complex interacting networks. In the ciliate Paramecium tetraurelia, at least two RNA interference (RNAi) mechanisms coexist, involving distinct but overlapping sets of protein factors and producing different types of short interfering RNAs (siRNAs). One is specifically triggered by high-copy transgenes, and the other by feeding cells with double-stranded RNA (dsRNA)-producing bacteria. In this study, we designed a forward genetic screen for mutants deficient in dsRNA-induced silencing, and a powerful method to identify the relevant mutations by whole-genome sequencing. We present a set of 47 mutant alleles for five genes, revealing two previously unknown RNAi factors: a novel Paramecium-specific protein (Pds1) and a Cid1-like nucleotidyl transferase. Analyses of allelic diversity distinguish non-essential and essential genes and suggest that the screen is saturated for non-essential, single-copy genes. We show that non-essential genes are specifically involved in dsRNA-induced RNAi while essential ones are also involved in transgene-induced RNAi. One of the latter, the RNA-dependent RNA polymerase RDR2, is further shown to be required for all known types of siRNAs, as well as for sexual reproduction. These results open the way for the dissection of the genetic complexity, interconnection, mechanisms and natural functions of RNAi pathways in P. tetraurelia. PMID:24860163

  18. Ciliary metachronal wave propagation on the compliant surface of Paramecium cells.

    PubMed

    Narematsu, Naoki; Quek, Raymond; Chiam, Keng-Hwee; Iwadate, Yoshiaki

    2015-12-01

    Ciliary movements in protozoa exhibit metachronal wave-like coordination, in which a constant phase difference is maintained between adjacent cilia. It is at present generally thought that metachronal waves require hydrodynamic coupling between adjacent cilia and the extracellular fluid. To test this hypothesis, we aspirated a Paramecium cell using a micropipette which completely sealed the surface of the cell such that no fluid could pass through the micropipette. Thus, the anterior and the posterior regions of the cell were hydrodynamically decoupled. Nevertheless, we still observed that metachronal waves continued to propagate from the anterior to the posterior ends of the cell, suggesting that in addition to hydrodynamic coupling, there are other mechanisms that can also transmit the metachronal waves. Such transmission was also observed in computational modeling where the fluid was fully decoupled between two partitions of a beating ciliary array. We also imposed cyclic stretching on the surface of live Paramecium cells and found that metachronal waves persisted in the presence of cyclic stretching. This demonstrated that, in addition to hydrodynamic coupling, a compliant substrate can also play a critical role in mediating the propagation of metachronal waves. © 2015 Wiley Periodicals, Inc.

  19. Epigenetic regulation of serotype expression antagonizes transcriptome dynamics in Paramecium tetraurelia.

    PubMed

    Cheaib, Miriam; Dehghani Amirabad, Azim; Nordström, Karl J V; Schulz, Marcel H; Simon, Martin

    2015-08-01

    Phenotypic variation of a single genotype is achieved by alterations in gene expression patterns. Regulation of such alterations depends on their time scale, where short-time adaptations differ from permanently established gene expression patterns maintained by epigenetic mechanisms. In the ciliate Paramecium, serotypes were described for an epigenetically controlled gene expression pattern of an individual multigene family. Paradoxically, individual serotypes can be triggered in Paramecium by alternating environments but are then stabilized by epigenetic mechanisms, thus raising the question to which extend their expression follows environmental stimuli. To characterize environmental adaptation in the context of epigenetically controlled serotype expression, we used RNA-seq to characterize transcriptomes of serotype pure cultures. The resulting vegetative transcriptome resource is first analysed for genes involved in the adaptive response to the altered environment. Secondly, we identified groups of genes that do not follow the adaptive response but show co-regulation with the epigenetically controlled serotype system, suggesting that their gene expression pattern becomes manifested by similar mechanisms. In our experimental set-up, serotype expression and the entire group of co-regulated genes were stable among environmental changes and only heat-shock genes altered expression of these gene groups. The data suggest that the maintenance of these gene expression patterns in a lineage represents epigenetically controlled robustness counteracting short-time adaptation processes. © The Author 2015. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.

  20. Intra-specific differentiation of Paramecium bursaria strains by molecular methods--preliminary studies.

    PubMed

    Greczek-Stachura, Magdalena; Tarcz, Sebastian; Przyboś, Ewa

    2010-01-01

    Ten strains of Paramecium bursaria and also P. caudatum, P. multimicronucleatum, P. tetraurelia strains (as outgroups) were characterized by using Random Amplified Polymorphic DNA (RAPD), Amplified Ribosomal DNA Restriction Analysis (ARDRA) and sequencing of the non-coding ribosomal internal transcribed spacer (ITS) regions. RAPD analysis revealed that all Paramecium bursaria strains possessed characteristic band patterns; there was a correlation between the degree of differentiation of DNA revealed by RAPD-fingerprinting and the geographic origin of a particular strain. ARDRA riboprinting (using a fragment of SSU-LSU rDNA, about 3085 bp) with restriction enzymes DraI, EcoRV, HhaI, HindIII, MspI, PstI distinguished groups of P. bursaria strains with characteristic band patterns originating from different sites. Comparison of the 550 bp ITS 1-5.8S-ITS2 fragment showed differentiation (0.9%) of the P. bursaria strains as three main groups of strains connected by site of origin in the constructed tree.

  1. Effect of horizontal strong static magnetic field on swimming behaviour of Paramecium caudatum

    NASA Astrophysics Data System (ADS)

    Fujiwara, Yoshihisa; Tomishige, Masahiko; Itoh, Yasuhiro; Fujiwara, Masao; Shibata, Naho; Kosaka, Toshikazu; Hosoya, Hiroshi; Tanimoto, Yoshifumi

    2006-05-01

    Effect of horizontal strong static magnetic field on swimming behaviour of Paramecium caudatum was studied by using a superconducting magnet. Around a centre of a round vessel, random swimming at 0 T and aligned swimming parallel to the magnetic field (MF) of 8 T were observed. Near a wall of the vessel, however, swimming round and round along the wall at 0 T and aligned swimming of turning at right angles upon collision with the wall, which was remarkable around 1-4 T, were detected. It was experimentally revealed that the former MF-induced parallel swimming at the vessel centre was caused physicochemically by the parallel magnetic orientation of the cell itself. From magnetic field dependence of the extent of the orientation, the magnetic susceptibility anisotropy (χ ∥-χ ⊥) was first obtained to be 3.4× 10-23 emu cell-1 at 298 K for Paramecium caudatum. The orientation of the cell was considered to result from the magnetic orientation of the cell membrane. On the other hand, although mechanisms of the latter swimming near the vessel wall regardless of the absence and presence of the magnetic field are unclear at present, these experimental results indicate that whether the cell exists near the wall alters the magnetic field effect on the swimming in the horizontal magnetic field.

  2. The SUMO pathway is developmentally regulated and required for programmed DNA elimination in Paramecium tetraurelia.

    PubMed

    Matsuda, Atsushi; Forney, James D

    2006-05-01

    Extensive genome-wide remodeling occurs during the formation of the somatic macronuclei from the germ line micronuclei in ciliated protozoa. This process is limited to sexual reproduction and includes DNA amplification, chromosome fragmentation, and the elimination of internal segments of DNA. Our efforts to define the pathways regulating these events revealed a gene encoding a homologue of ubiquitin activating enzyme 2 (UBA2) that is upregulated at the onset of macronuclear development in Paramecium tetraurelia. Uba2 enzymes are known to activate the protein called small ubiquitin-related modifier (SUMO) that is covalently attached to target proteins. Consistent with this relationship, Northern analysis showed increased abundance of SUMO transcripts during sexual reproduction in Paramecium. RNA interference (RNAi) against UBA2 or SUMO during vegetative growth had little effect on cell survival or fission rates. In contrast, RNAi of mating cells resulted in failure to form a functional macronucleus. Despite normal amplification of the genome, excision of internal eliminated sequences was completely blocked. Additional experiments showed that the homologous UBA2 and SUMO genes in Tetrahymena thermophila are also upregulated during conjugation. These results provide evidence for the developmental regulation of the SUMO pathway in ciliates and suggest a key role for the pathway in controlling genome remodeling.

  3. Effects of space balloon flights on reproductive activity in Paramecium aurelia.

    PubMed

    Planel, H; Soleilhavoup, J P; Croute, F

    1975-01-01

    Post autogamous Paramecium aurelia cultures were placed in hermetic containers, including a heating device with accuracy kept around +/- 0.1 degrees C. Kinetics of cellular growth was determined by cell count, after recovery, on in-flight cultures and ground control cultures. Dosimetry was performed by thermoluminescent detectors (CaSO4 activated with dysprosium). Flight durations of maximum altitude (ceiling) ranged between 48 min and 15 hours (repeated flights). Conclusions are as follows: short flights result in a secondary stimulating effect, shown by post-flight increase of the growth rate (total dose above 2 mrads); long flights or repeated flights are accompanied by a decrease in growth rate (total dose ranging from 2 to 6 mrads); in the stimulation experiments, cell counts performed immediately after flight permit identification of a temporary decrease of growth rate. The biphasic character of the biological response after flights may be due to an ionization phenomena induced by cosmic rays. Indeed, the temporary drop of growth rate is not observed after recovery if the cells are subcultured in fresh medium and left on the earth's surface. We observe, on the contrary, an increase in growth rate. These findings confirm the great sensitivity of Paramecium aurelia to very low doses of ionizing radiations and demonstrate the biological effect of cosmic radiation.

  4. The SUMO Pathway Is Developmentally Regulated and Required for Programmed DNA Elimination in Paramecium tetraurelia† ‡

    PubMed Central

    Matsuda, Atsushi; Forney, James D.

    2006-01-01

    Extensive genome-wide remodeling occurs during the formation of the somatic macronuclei from the germ line micronuclei in ciliated protozoa. This process is limited to sexual reproduction and includes DNA amplification, chromosome fragmentation, and the elimination of internal segments of DNA. Our efforts to define the pathways regulating these events revealed a gene encoding a homologue of ubiquitin activating enzyme 2 (UBA2) that is upregulated at the onset of macronuclear development in Paramecium tetraurelia. Uba2 enzymes are known to activate the protein called small ubiquitin-related modifier (SUMO) that is covalently attached to target proteins. Consistent with this relationship, Northern analysis showed increased abundance of SUMO transcripts during sexual reproduction in Paramecium. RNA interference (RNAi) against UBA2 or SUMO during vegetative growth had little effect on cell survival or fission rates. In contrast, RNAi of mating cells resulted in failure to form a functional macronucleus. Despite normal amplification of the genome, excision of internal eliminated sequences was completely blocked. Additional experiments showed that the homologous UBA2 and SUMO genes in Tetrahymena thermophila are also upregulated during conjugation. These results provide evidence for the developmental regulation of the SUMO pathway in ciliates and suggest a key role for the pathway in controlling genome remodeling. PMID:16682458

  5. Insights into Three Whole-Genome Duplications Gleaned from the Paramecium caudatum Genome Sequence

    PubMed Central

    McGrath, Casey L.; Gout, Jean-Francois; Doak, Thomas G.; Yanagi, Akira; Lynch, Michael

    2014-01-01

    Paramecium has long been a model eukaryote. The sequence of the Paramecium tetraurelia genome reveals a history of three successive whole-genome duplications (WGDs), and the sequences of P. biaurelia and P. sexaurelia suggest that these WGDs are shared by all members of the aurelia species complex. Here, we present the genome sequence of P. caudatum, a species closely related to the P. aurelia species group. P. caudatum shares only the most ancient of the three WGDs with the aurelia complex. We found that P. caudatum maintains twice as many paralogs from this early event as the P. aurelia species, suggesting that post-WGD gene retention is influenced by subsequent WGDs and supporting the importance of selection for dosage in gene retention. The availability of P. caudatum as an outgroup allows an expanded analysis of the aurelia intermediate and recent WGD events. Both the Guanine+Cytosine (GC) content and the expression level of preduplication genes are significant predictors of duplicate retention. We find widespread asymmetrical evolution among aurelia paralogs, which is likely caused by gradual pseudogenization rather than by neofunctionalization. Finally, cases of divergent resolution of intermediate WGD duplicates between aurelia species implicate this process acts as an ongoing reinforcement mechanism of reproductive isolation long after a WGD event. PMID:24840360

  6. Functional imaging of living Paramecium by means of confocal and two-photon excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Diaspro, Alberto; Fronte, Paola; Raimondo, Marco; Fato, Marco; DeLeo, Gianluca; Beltrame, Francesco; Cannone, Fabio; Chirico, Giberto; Ramoino, Paola

    2002-05-01

    Confocal and Two-photon excitation laser scanning microscopy allow gathering three-dimensional and temporal information from biological systems exploiting fluorescence labeling and autofluorescence properties. In this work we study biological events linked to functionality in Paramecium primaurelia. The internalization of material in ciliated one-celled organisms (protozoa) occurs via different mechanisms, even if most of nutrients, particulate or not, is taken up by food vacuoles formed at the bottom of the oral cavity. The endocytosis of small-sized molecules occurs at the parasomal sacs, located next the ciliar basal bodies. Vital fluorescent dyes (BSA-FITC, WGA-FITC, dextran-Texas Red, cholesteryl-Bodipy) and autofluorescence were used to study formation, movement, and fusion of vesicles during endocytosis and phagocytosis of Paramecium primaurelia. By immobilizing living cells pulsed with food vacuole and endosome markers at successive times after chasing in unlabeled medium, the intracellular movement and fusion of food vacuoles and of endosomes were visualized. A temporal analysis of fluorescence images and the false-color technique were used. Starting from time series or 3D data sets composite images were generated by associating with each originally acquired image a different color corresponding to each sampling point in time and along the z-axis. Second Harmonic Generation Imaging attempts are also outlined.

  7. New stands of species of the Paramecium aurelia complex in Africa and Europe.

    PubMed

    Rautian, Maria; Przyboś, Ewa; Surmacz, Marta; Lebedeva, Natalia

    2014-01-01

    The relevance of geographical distribution and the roles of dispersal and spatial isolation during the speciation of microorganisms are nowadays of great interest. The Paramecium aurelia species complex is a perfect model system to explore these questions given its long history as a study subject and broad distribution. However, the world-wide distribution of the Paramecium aurelia complex (Ciliophora, Protista) still needs study, e.g., sampling in the southern hemisphere has been quite limited, while Europe has been investigated for years, with the majority of aurelia species isolated from here. Recently, new stands of species of the P. aurelia complex were found in southern Europe (Malta, Bulgaria, Cyprus) and in the Czech Republic (P. primaurelia, P. triaurelia, P. octaurelia). In Africa (Republic of South Africa), new stands of P. primaurelia, P. triaurelia, and P. octaurelia were found. Interestingly, the rare species P. triaurelia, and P. octaurelia were found to co-occur both in South Africa (SA 13) and the Czech Republic (CKV 8). Newly established strains were identified to species by crossing with the test strains (the reference strains for the particular species).

  8. Orientation of paramecium swimming in a static magnetic field: Dependence on membrane lipid fluidity.

    PubMed

    Nakaoka, Yasuo; Itoh, Junya; Shimizu, Kikuo

    2011-01-01

    We studied the swimming orientation of the ciliated protozoan Paramecium aurelia in a static magnetic field (0.78 T). P. aurelia is a complex of species termed syngens, whose cell morphology appears similar on microscopic examination. In the magnetic field, the cells of some syngens gradually changed their swimming orientation so that they were swimming perpendicular or parallel to the magnetic field, although such sensitivity to magnetic fields differs between syngens. When the temperature of the cell suspension was raised, the magnetic sensitivity of the cells was decreased. On the other hand, when the cells were cultured beforehand at a high temperature, their magnetic sensitivity was increased. These results raise the possibility that membrane lipid fluidity, which is inversely proportional to the membrane lipid order, contributes to the magnetic orientation of syngens. In this study, measurements of membrane lipid fluidity obtained using fluorescence image analysis with the lipophilic dye, laurdan (6-lauroyl-2-dimethylaminonaphtalene), showed that the degree of membrane lipid fluidity was correlated with the differences in magnetic orientation between syngens. That is, the syngens with decreased membrane fluidity showed an increased degree of magnetic orientation. Therefore, the membrane lipid order is a key factor in the magnetic orientation of Paramecium swimming.

  9. FOOD VACUOLE MEMBRANE GROWTH WITH MICROTUBULE-ASSOCIATED MEMBRANE TRANSPORT IN PARAMECIUM

    PubMed Central

    Allen, Richard D.

    1974-01-01

    Evidence from a morphological study of the oral apparatus of Paramecium caudatum using electron microscope techniques have shown the existence of an elaborate structural system which is apparently designed to recycle digestive-vacuole membrane. Disk-shaped vesicles are filtered out of the cytoplasm by a group of microtubular ribbons. The vesicles, after being transported to the cytostome-cytopharynx region in association with these ribbons, accumulate next to the cytopharynx before they become fused with the cytopharyngeal membrane. This fusion allows the nascent food vacuole to grow and increase its membrane surface area. The morphology of this cytostome-cytopharynx region is described in detail and illustrated with a three-dimensional drawing of a portion of this region and a clay sculpture of the oral apparatus of Paramecium. Evidence from the literature for the transformation of food vacuole membrane into disk-shaped vesicles both from condensing food vacuoles in the endoplasm and from egested food vacuoles at the cytoproct is presented. This transformation would complete a system of digestive vacuole membrane recycling. PMID:4373478

  10. A large multigene family codes for the polypeptides of the crystalline trichocyst matrix in Paramecium.

    PubMed Central

    Madeddu, L; Gautier, M C; Vayssié, L; Houari, A; Sperling, L

    1995-01-01

    The secretory granules (trichocysts) of Paramecium are characterized by a highly constrained shape that reflects the crystalline organization of their protein contents. Yet the crystalline trichocyst content is composed not of a single protein but of a family of related polypeptides that derive from a family of precursors by protein processing. In this paper we show that a multigene family, of unusually large size for a unicellular organism, codes for these proteins. The family is organized in subfamilies; each subfamily codes for proteins with different primary structures, but within the subfamilies several genes code for nearly identical proteins. For one subfamily, we have obtained direct evidence that the different members are coexpressed. The three subfamilies we have characterized are located on different macronuclear chromosomes. Typical 23-29 nucleotide Paramecium introns are found in one of the regions studied and the intron sequences are more variable than the surrounding coding sequences, providing gene-specific markers. We suggest that this multigene family may have evolved to assure a microheterogeneity of structural proteins necessary for morphogenesis of a complex secretory granule core with a constrained shape and dynamic properties: genetic analysis has shown that correct assembly of the crystalline core is necessary for trichocyst function. Images PMID:7579685

  11. Protein phosphatase and kinase activities possibly involved in exocytosis regulation in Paramecium tetraurelia.

    PubMed

    Kissmehl, R; Treptau, T; Hofer, H W; Plattner, H

    1996-07-01

    In Paramecium tetraurelia cells synchronous exocytosis induced by aminoethyldextran (AED) is accompanied by an equally rapid dephosphorylation of a 63 kDa phosphoprotein (PP63) within 80 ms. In vivo, rephosphorylation occurs within a few seconds after AED triggering. In homogenates (P)P63 can be solubilized in all three phosphorylation states (phosphorylated, dephosphorylated and rephosphorylated) and thus tested in vitro. By using chelators of different divalent cations, de- and rephosphorylation of PP63 and P63 respectively can be achieved by an endogenous protein phosphatase/kinase system. Dephosphorylation occurs in the presence of EDTA, whereas in the presence of EGTA this was concealed by phosphorylation by endogenous kinase(s), thus indicating that phosphorylation of P63 is calcium-independent. Results obtained with protein phosphatase inhibitors (okadaic acid, calyculin A) allowed us to exclude a protein serine/threonine phosphatase of type I (with selective sensitivity in Paramecium). Protein phosphatase 2C is also less likely to be a candidate because of its requirement for high Mg2+ concentrations. According to previous evidence a protein serine/threonine phosphatase of type 2B (calcineurin; CaN) is possibly involved. We have now found that bovine brain CaN dephosphorylates PP63 in vitro. Taking into account the specific requirements of this phosphatase in vitro, with p-nitrophenyl phosphate as a substrate, we have isolated a cytosolic phosphatase of similar characteristics by combined preparative gel electrophoresis and affinity-column chromatography. In Paramecium this phosphatase also dephosphorylates PP63 in vitro (after 32P labelling in vivo). Using various combinations of ion exchange, affinity and hydrophobic interaction chromatography we have also isolated three different protein kinases from the soluble fraction, i.e. a cAMP-dependent protein kinase (PKA), a cGMP-dependent protein kinase (PKG) and a casein kinase. Among the kinases tested, PKA

  12. A 63 kDa phosphoprotein undergoing rapid dephosphorylation during exocytosis in Paramecium cells shares biochemical characteristics with phosphoglucomutase.

    PubMed

    Treptau, T; Kissmehl, R; Wissmann, J D; Plattner, H

    1995-07-15

    We have enriched phosphoglucomutase (PGM; EC 5.4.2.2) approximately 20-fold from Paramecium tetraurelia cells by combined fractional precipitation with (NH4)2SO4, gel filtration and anion-exchange chromatography yielding two PGM peaks. Several parameters affecting PGM enzymic activity, molecular mass and pI were determined. Phosphorylation studies were done with isolated endogenous protein kinases. Like the 63 kDa phosphoprotein PP63, which is dephosphorylated within 80 ms during synchronous trichocyst exocytosis [Höhne-Zell, Knoll, Riedel-Gras, Hofer and Plattner (1992) Biochem. J. 286, 843-849], PGM has a molecular mass of 63 kDa and forms of identical pI. Since mammalian PGM activity depends on the presence of glucose 1,6-bisphosphate (Glc-1,6-P2) (which is lost during anion-exchange chromatography), we analysed this aspect with Paramecium PGM. In this case PGM activity was shown not to be lost, due to p-nitrophenyl phosphate-detectable phosphatase(s) (which we have separated from PGM), but also due to loss of Glc-1,6-P2. Like PGM from various vertebrate species, PGM activity from Paramecium can be fully re-established by addition of Glc-1,6-P2 at 10 nM, and it is also stimulated by bivalent cations and insensitive to chelating or thiol reagents. The PGM which we have isolated can be phosphorylated by endogenous cyclic-GMP-dependent protein kinase or by endogenous casein kinase. This results in three phosphorylated bands of identical molecular mass and pI values, as we have shown to occur with PP63 after phosphorylation in vivo (forms with pI 6.05, 5.95, 5.85). In ELISA, antibodies raised against PGM from rabbit skeletal muscle were reactive not only with original PGM but also with PGM fractions from Paramecium. Therefore, PGM and PP63 seem to be identical with regard to widely different parameters, i.e. co-elution by chromatography, molecular mass, phosphorylation by the two protein kinases tested, pI values of isoforms, and immuno-binding. Recent claims that

  13. A 63 kDa phosphoprotein undergoing rapid dephosphorylation during exocytosis in Paramecium cells shares biochemical characteristics with phosphoglucomutase.

    PubMed Central

    Treptau, T; Kissmehl, R; Wissmann, J D; Plattner, H

    1995-01-01

    We have enriched phosphoglucomutase (PGM; EC 5.4.2.2) approximately 20-fold from Paramecium tetraurelia cells by combined fractional precipitation with (NH4)2SO4, gel filtration and anion-exchange chromatography yielding two PGM peaks. Several parameters affecting PGM enzymic activity, molecular mass and pI were determined. Phosphorylation studies were done with isolated endogenous protein kinases. Like the 63 kDa phosphoprotein PP63, which is dephosphorylated within 80 ms during synchronous trichocyst exocytosis [Höhne-Zell, Knoll, Riedel-Gras, Hofer and Plattner (1992) Biochem. J. 286, 843-849], PGM has a molecular mass of 63 kDa and forms of identical pI. Since mammalian PGM activity depends on the presence of glucose 1,6-bisphosphate (Glc-1,6-P2) (which is lost during anion-exchange chromatography), we analysed this aspect with Paramecium PGM. In this case PGM activity was shown not to be lost, due to p-nitrophenyl phosphate-detectable phosphatase(s) (which we have separated from PGM), but also due to loss of Glc-1,6-P2. Like PGM from various vertebrate species, PGM activity from Paramecium can be fully re-established by addition of Glc-1,6-P2 at 10 nM, and it is also stimulated by bivalent cations and insensitive to chelating or thiol reagents. The PGM which we have isolated can be phosphorylated by endogenous cyclic-GMP-dependent protein kinase or by endogenous casein kinase. This results in three phosphorylated bands of identical molecular mass and pI values, as we have shown to occur with PP63 after phosphorylation in vivo (forms with pI 6.05, 5.95, 5.85). In ELISA, antibodies raised against PGM from rabbit skeletal muscle were reactive not only with original PGM but also with PGM fractions from Paramecium. Therefore, PGM and PP63 seem to be identical with regard to widely different parameters, i.e. co-elution by chromatography, molecular mass, phosphorylation by the two protein kinases tested, pI values of isoforms, and immuno-binding. Recent claims that

  14. Identification and cloning of first cadmium metallothionein like gene from locally isolated ciliate, Paramecium sp.

    PubMed

    Shuja, Rukhsana Nighat; Shakoori, Abdul Rauf

    2009-03-01

    First cadmium metallothionein like gene PMCd1 of a ciliate, Paramecium sp., isolated from industrial wastewater has been cloned and sequenced. PMCd1 is an intronless gene, encoding 612 nucleotides, with TAA coding for glutamine. The coding region of PMCd1 comprises 203 amino acids, including 37 cysteine residues with a conserved structural pattern in the form of recurring structural motifs, arranged in 17 x-cys-x-y-cys-x, 1 x-cys-cys-x and x-cys-x contexts. Both, the deduced amino acids and nucleotide sequence differ, not only from other animal metallothioneins (MTs), but also from the previously characterized Tetrahymena Cu and Cd-MTs. The translated protein of PMCd1 contains conserved cysteine residues, peculiar characteristic of stress inducible metallothionein genes of ciliates and other groups of organisms.

  15. Attempts to retreat from a dead-ended long capillary by backward swimming in Paramecium

    PubMed Central

    Kunita, Itsuki; Kuroda, Shigeru; Ohki, Kaito; Nakagaki, Toshiyuki

    2014-01-01

    We have observed how the ciliate Paramecium attempts to retreat from the dead-end of a long capillary that is too narrow for turning. After many trial-and-error episodes of short-term backward swimming (SBS), which is the conventional avoidance behavior exhibited in free swimming when an obstacle is faced, long-term backward swimming (LBS) that lasted five to ten times longer was developed. LBS may have a beneficial effect for complete withdrawal from the capillary space, although in our experiment it was impossible for the organism to do so due to the capillary length. In order to identify a physically possible mechanism for LBS, we propose model equations for the membrane potential of Hodgkin–Huxley type, which describe the control of ciliary movement. The physiological implications and physical mechanism of the development of LBS are discussed. PMID:24966852

  16. Stomatogenesis during sexual and asexual reproduction in an amicronucleate strain of Paramecium caudatum.

    PubMed

    Mikami, K

    1979-04-01

    Amicronucleate cells of Paramecium caudatum, whose micronuclei have been artifically removed by micropipetting, are characterized by the appearance of a deciliated area at the posterior part of the buccal opening. These cells form food vacuoles at a slightly lower rate than micronucleate cells. Their mean interfission time is longer than that in micronucleates. The exconjugants of amicronucleate cells can not form food vacuoles and eventually die witout fission, though conjugation proceeds normally in them as well as in their micronucleate mate. The oral apparatus of amicronucleate exconjugants seems to be shallower than that of micronucleates. The membranellar cilia, therefore, can be seen through the buccal overture by scanning electron microscope. The results obtained from the cross of micronucleate and amicronucleate strains and from the induction of autogamy in amicronucleate strains suggest that the micronucleus has a primary role in developing the normal oral apparatus after nuclear reorganization.

  17. Effect of Cypermethrin on the Growth of Ciliate Protozoan Paramecium caudatum

    PubMed Central

    Dutta, Joydeep

    2015-01-01

    Objective: The objective of this study is to assess the effect of cypermethrin on the growth of ciliate protozoan Paramecium caudatum. Materials and Methods: Monoxenic culture of P. caudatum, were exposed to different doses (0.01, 0.05, 0.1, 0.15, and 0.2 µg/L) of cypermethrin along with control for 24, 48, 72, and 96 h time interval. The total numbers of live and dead cells were counted after trypan blue staining in Neubauer hemocytometer. Results: Marked decrease in the number of living cells with the increase in the concentration of cypermethrin and with increasing exposure time intervals was recorded. Conclusion: The results indicate that cypermethrin is toxic to P. caudatum even at low concentrations when it enters in the aquatic system through runoff. PMID:26862268

  18. A comparative hybridization analysis of yeast DNA with Paramecium parafusin- and different phosphoglucomutase-specific probes.

    PubMed

    Wyroba, E; Satir, B H

    2000-01-01

    Molecular probes designed for the parafusin (PFUS), the Paramecium exocytic-sensitive phosphoglycoprotein, gave distinct hybridization patterns in Saccharomyces cerevisiae genomic DNA when compared with different phosphoglucomutase specific probes. These include two probes identical to segments of yeast phosphoglucomutase (PGM) genes 1 and 2. Neither of the PGM probes revealed the 7.4 and 5.9 kb fragments in Bgl II-cut yeast DNA digest detected with the 1.6 kb cloned PFUS cDNA and oligonucleotide constructed to the PFUS region (insertion 3--I-3) not found in other species. PCR amplification with PFUS-specific primers generated yeast DNA-species of the predicted molecular size which hybridized to the I-3 probe. A search of the yeast genome database produced an unassigned nucleotide sequence that showed 55% identity to parafusin gene and 37% identity to PGM2 (the major isoform of yeast phosphoglucomutase) within the amplified region.

  19. Longevity in space; experiment on the life span of Paramecium cell clone in space.

    PubMed

    Mogami, Y; Tokunaga, N; Baba, S A

    1999-01-01

    Life span is the most interesting and also the most important biologically relevant time to be investigated on the space station. As a model experiment, we proposed an investigation to assess the life span of clone generation of the ciliate Paramecium. In space, clone generation will be artificially started by conjugation or autogamy, and the life span of the cell populations in different gravitational fields (microgravity and onboard 1 x g control) will be precisely assessed in terms of fission age as compared with the clock time. In order to perform the space experiment including long-lasting culture and continuous measurement of cell division, we tested the methods of cell culture and of cell-density measurement, which will be available in closed environments under microgravity. The basic design of experimental hardware and a preliminary result of the cultivation procedure are described. c1999 COSPAR. Published by Elsevier Science Ltd.

  20. The Impact of UV Radiation on Paramecium Populations from Alpine Lakes.

    PubMed

    Kammerlander, Barbara; Tartarotti, Barbara; Sonntag, Bettina

    2017-08-21

    Paramecium populations from a clear and a glacier-fed turbid alpine lake were exposed to solar simulated ultraviolet (UVR) and photosynthetically active radiation (PAR) at 8 and 15 °C. The ciliates were tested for DNA damage (comet assay), behavioral changes, and mortality after UVR + PAR exposure. High DNA damage levels (~58% tail DNA) and abnormal swimming behavior were observed, although no significant changes in cell numbers were found irrespective of the lake origin (clear, turbid), and temperatures. We conclude that environmental stressors such as UVR and their effects may influence the adaptation of ciliates living in alpine lakes. © 2017 The Author(s) Journal of Eukaryotic Microbiology published by Wiley Periodicals, Inc. on behalf of International Society of Protistologists.

  1. Attempts to retreat from a dead-ended long capillary by backward swimming in Paramecium.

    PubMed

    Kunita, Itsuki; Kuroda, Shigeru; Ohki, Kaito; Nakagaki, Toshiyuki

    2014-01-01

    We have observed how the ciliate Paramecium attempts to retreat from the dead-end of a long capillary that is too narrow for turning. After many trial-and-error episodes of short-term backward swimming (SBS), which is the conventional avoidance behavior exhibited in free swimming when an obstacle is faced, long-term backward swimming (LBS) that lasted five to ten times longer was developed. LBS may have a beneficial effect for complete withdrawal from the capillary space, although in our experiment it was impossible for the organism to do so due to the capillary length. In order to identify a physically possible mechanism for LBS, we propose model equations for the membrane potential of Hodgkin-Huxley type, which describe the control of ciliary movement. The physiological implications and physical mechanism of the development of LBS are discussed.

  2. Mendelian and non-mendelian mutations affecting surface antigen expression in Paramecium tetraurelia

    SciTech Connect

    Epstein, L.M.; Forney, J.D.

    1984-08-01

    A screening procedure was devised for the isolation of X-ray-induced mutations affecting the expression of the A immobilization antigen (i-antigen) in Paramecium tetraurelia. Two of the mutations isolated by this procedure proved to be in modifier genes. The two genes are unlinked to each other and unlinked to the structural A i-antigen gene. These are the first modifier genes identified in a Paramecium sp. that affect surface antigen expression. Another mutation was found to be a deletion of sequences just downstream from the A i-antigen gene. In cells carrying this mutation, the A i-antigen gene lies in close proximity to the end of a macronuclear chromosome. The expression of the A i-antigen is not affected in these cells, demonstrating that downstream sequences are not important for the regulation and expression of the A i-antigen gene. A stable cell line was also recovered which shows non-Mendelian inheritance of a macronuclear deletion of the A i-antigen gene. This mutant does not contain the gene in its macronucleus, but contains a complete copy of the gene in its micronucleus. In the cytoplasm of wild-type animals, the micronuclear gene is included in the developing macronucleus; in the cytoplasm of the mutant, the incorporation of the A i-antigen gene into the macronucleus is inhibited. This is the first evidence that a mechanism is available in ciliates to control the expression of a gene by regulating its incorporation into developing macronuclei.

  3. Temporal variation in temperature determines disease spread and maintenance in Paramecium microcosm populations

    PubMed Central

    Duncan, Alison B.; Fellous, Simon; Kaltz, Oliver

    2011-01-01

    The environment is rarely constant and organisms are exposed to temporal and spatial variations that impact their life histories and inter-species interactions. It is important to understand how such variations affect epidemiological dynamics in host–parasite systems. We explored effects of temporal variation in temperature on experimental microcosm populations of the ciliate Paramecium caudatum and its bacterial parasite Holospora undulata. Infected and uninfected populations of two P. caudatum genotypes were created and four constant temperature treatments (26°C, 28°C, 30°C and 32°C) compared with four variable treatments with the same mean temperatures. Variable temperature treatments were achieved by alternating populations between permissive (23°C) and restrictive (35°C) conditions daily over 30 days. Variable conditions and high temperatures caused greater declines in Paramecium populations, greater fluctuations in population size and higher incidence of extinction. The additional effect of parasite infection was additive and enhanced the negative effects of the variable environment and higher temperatures by up to 50 per cent. The variable environment and high temperatures also caused a decrease in parasite prevalence (up to 40%) and an increase in extinction (absence of detection) (up to 30%). The host genotypes responded similarly to the different environmental stresses and their effect on parasite traits were generally in the same direction. This work provides, to our knowledge, the first experimental demonstration that epidemiological dynamics are influenced by environmental variation. We also emphasize the need to consider environmental variance, as well as means, when trying to understand, or predict population dynamics or range. PMID:21450730

  4. A forward genetic screen reveals essential and non-essential RNAi factors in Paramecium tetraurelia.

    PubMed

    Marker, Simone; Carradec, Quentin; Tanty, Véronique; Arnaiz, Olivier; Meyer, Eric

    2014-06-01

    In most eukaryotes, small RNA-mediated gene silencing pathways form complex interacting networks. In the ciliate Paramecium tetraurelia, at least two RNA interference (RNAi) mechanisms coexist, involving distinct but overlapping sets of protein factors and producing different types of short interfering RNAs (siRNAs). One is specifically triggered by high-copy transgenes, and the other by feeding cells with double-stranded RNA (dsRNA)-producing bacteria. In this study, we designed a forward genetic screen for mutants deficient in dsRNA-induced silencing, and a powerful method to identify the relevant mutations by whole-genome sequencing. We present a set of 47 mutant alleles for five genes, revealing two previously unknown RNAi factors: a novel Paramecium-specific protein (Pds1) and a Cid1-like nucleotidyl transferase. Analyses of allelic diversity distinguish non-essential and essential genes and suggest that the screen is saturated for non-essential, single-copy genes. We show that non-essential genes are specifically involved in dsRNA-induced RNAi while essential ones are also involved in transgene-induced RNAi. One of the latter, the RNA-dependent RNA polymerase RDR2, is further shown to be required for all known types of siRNAs, as well as for sexual reproduction. These results open the way for the dissection of the genetic complexity, interconnection, mechanisms and natural functions of RNAi pathways in P. tetraurelia. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. Paramecium caudatum enhances transmission and infectivity of Mycobacterium marinum and Mycobacterium chelonae in zebrafish (Danio rerio)

    PubMed Central

    Peterson, Tracy S.; Ferguson, Jayde A.; Watral, Virginia G.; Mutoji, K. Nadine; Ennis, Don G.; Kent, Michael L.

    2014-01-01

    Mycobacterial infections in laboratory zebrafish (Danio rerio) are common and widespread in research colonies. Mycobacteria within free living amoebae have been shown to be transmission vectors for mycobacteriosis. Paramecium caudatum are commonly used as a first food for zebrafish, and we investigated this ciliate’s potential to serve as a vector of Mycobacterium marinum and M. chelonae. The ability of live P. caudatum to transmit these mycobacteria to larval, juvenile and adult zebrafish was evaluated. Infections were defined by histologic observation of granulomas containing acid-fast bacteria in extraintestinal locations. In both experiments, fish fed paramecia containing mycobacteria became infected at a higher incidence than controls. Larvae (exposed at 4 days post hatch) fed paramecia with M. marinum exhibited an incidence of 30% (24/80) and juveniles (exposed at 21 days post hatch) showed 31% incidence (14/45). Adult fish fed a gelatin food matrix containing mycobacteria within paramecia or mycobacteria alone for 2 wk resulted in infections when examined 8 wk after exposure as follows: M. marinum OSU 214 47% (21/45), M. marinum CH 47% (9/19), M. chelonae 38% (5/13). In contrast, fish feed mycobacteria alone in this diet did not become infected, except for 2 fish (5%) in the M. marinum OSU 214 low dose group. These results demonstrate that Paramecium caudatum can act as a vector for mycobacteria. This provides a useful animal model for evaluation of natural mycobacterial infections and demonstrates the possibility of mycobacterial transmission in zebrafish facilities via contaminated paramecia cultures. PMID:24192000

  6. Novel Insights into the Development and Function of Cilia Using the Advantages of the Paramecium Cell and Its Many Cilia

    PubMed Central

    Yano, Junji; Valentine, Megan S.; Van Houten, Judith L.

    2015-01-01

    Paramecium species, especially P. tetraurelia and caudatum, are model organisms for modern research into the form and function of cilia. In this review, we focus on the ciliary ion channels and other transmembrane proteins that control the beat frequency and wave form of the cilium by controlling the signaling within the cilium. We put these discussions in the context of the advantages that Paramecium brings to the understanding of ciliary motility: mutants for genetic dissections of swimming behavior, electrophysiology, structural analysis, abundant cilia for biochemistry and modern proteomics, genomics and molecular biology. We review the connection between behavior and physiology, which allows the cells to broadcast the function of their ciliary channels in real time. We build a case for the important insights and advantages that this model organism continues to bring to the study of cilia. PMID:26230712

  7. 808-nm laser therapy with a flat-top handpiece photobiomodulates mitochondria activities of Paramecium primaurelia (Protozoa).

    PubMed

    Amaroli, Andrea; Ravera, Silvia; Parker, Steven; Panfoli, Isabella; Benedicenti, Alberico; Benedicenti, Stefano

    2016-05-01

    Photobiomodulation is proposed as a non-linear process, and only low-level laser therapy (LLLT) is assumed to stimulate exposed cells, whereas high powered laser and fluences can cause negative effects, exhausting the cell's energy reserve as a consequence of excessive photon-based stimulation. In our work, we investigated and compared the effects of 808-nm diode laser (CW) with a new flat-top handpiece. To this purpose, we tested the photobiomodulation effects of 1 and 3 J/cm(2) fluence, both generated by 100 mW or 1 W of laser power and of 64 J/cm(2) of fluence generated by 100 mW, 1 W, 1.5 W or 2 W, as expressed through oxygen consumption and ATP synthesis of Paramecium. Data collected indicates the incremental consumption of oxygen through irradiation with 3 J/cm(2)-100 mW or 64 J/cm(2)-1 W correlates with an increase in Paramecium ATP synthesis. The Paramecium respiration was inhibited by fluences 64 J/cm(2)-100 mW or 64 J/cm(2)-2 W and was followed by a decrease in the endogenous ATP concentration. The 1 J/cm(2)-100 mW or 1 W and 3 J/cm(2)-1 W did not affect mitochondrial activity. The results show that the fluence of 64 J/cm(2)-1 W more than the 3 J/cm(2)-100 mW causes greater efficiency in Paramecium mitochondria respiratory chain activity. Our results suggest that thanks to flat-top handpiece we used, high fluences by high-powered laser have to be reconsidered as an effective and non-invasive therapy. Possible associated benefits of deeper tissue penetration would increase treatment effectiveness and reduced irradiation time.

  8. Influence on cell proliferation of background radiation or exposure to very low, chronic gamma radiation. [Paramecium tetraurelia; Synechococcus lividus

    SciTech Connect

    Planel, H.; Soleilhavoup, J.P.; Tixador, R.; Richoilley, G.; Conter, A.; Croute, F.; Caratero, C.; Gaubin, Y.

    1987-05-01

    Investigations carried out on the protozoan Paramecium tetraurelia and the cyanobacteria Synechococcus lividus, which were shielded against background radiation or exposed to very low doses of gamma radiation, demonstrated that radiation can stimulate the proliferation of these two single-cell organisms. Radiation hormesis depends on internal factors (age of starting cells) and external factors (lighting conditions). The stimulatory effect occurred only in a limited range of doses and disappeared for dose rates higher than 50 mGy/y.

  9. Cross-study analysis of genomic data defines the ciliate multigenic epiplasmin family: strategies for functional analysis in Paramecium tetraurelia

    PubMed Central

    Damaj, Raghida; Pomel, Sébastien; Bricheux, Geneviève; Coffe, Gérard; Viguès, Bernard; Ravet, Viviane; Bouchard, Philippe

    2009-01-01

    Background The sub-membranous skeleton of the ciliate Paramecium, the epiplasm, is composed of hundreds of epiplasmic scales centered on basal bodies, and presents a complex set of proteins, epiplasmins, which belong to a multigenic family. The repeated duplications observed in the P. tetraurelia genome present an interesting model of the organization and evolution of a multigenic family within a single cell. Results To study this multigenic family, we used phylogenetic, structural, and analytical transcriptional approaches. The phylogenetic method defines 5 groups of epiplasmins in the multigenic family. A refined analysis by Hydrophobic Cluster Analysis (HCA) identifies structural characteristics of 51 epiplasmins, defining five separate groups, and three classes. Depending on the sequential arrangement of their structural domains, the epiplasmins are defined as symmetric, asymmetric or atypical. The EST data aid in this classification, in the identification of putative regulating sequences such as TATA or CAAT boxes. When specific RNAi experiments were conducted using sequences from either symmetric or asymmetric classes, phenotypes were drastic. Local effects show either disrupted or ill-shaped epiplasmic scales. In either case, this results in aborted cell division. Using structural features, we show that 4 epiplasmins are also present in another ciliate, Tetrahymena thermophila. Their affiliation with the distinctive structural groups of Paramecium epiplasmins demonstrates an interspecific multigenic family. Conclusion The epiplasmin multigenic family illustrates the history of genomic duplication in Paramecium. This study provides a framework which can guide functional analysis of epiplasmins, the major components of the membrane skeleton in ciliates. We show that this set of proteins handles an important developmental information in Paramecium since maintenance of epiplasm organization is crucial for cell morphogenesis. PMID:19493334

  10. Cross-study analysis of genomic data defines the ciliate multigenic epiplasmin family: strategies for functional analysis in Paramecium tetraurelia.

    PubMed

    Damaj, Raghida; Pomel, Sébastien; Bricheux, Geneviève; Coffe, Gérard; Viguès, Bernard; Ravet, Viviane; Bouchard, Philippe

    2009-06-03

    The sub-membranous skeleton of the ciliate Paramecium, the epiplasm, is composed of hundreds of epiplasmic scales centered on basal bodies, and presents a complex set of proteins, epiplasmins, which belong to a multigenic family. The repeated duplications observed in the P. tetraurelia genome present an interesting model of the organization and evolution of a multigenic family within a single cell. To study this multigenic family, we used phylogenetic, structural, and analytical transcriptional approaches. The phylogenetic method defines 5 groups of epiplasmins in the multigenic family. A refined analysis by Hydrophobic Cluster Analysis (HCA) identifies structural characteristics of 51 epiplasmins, defining five separate groups, and three classes. Depending on the sequential arrangement of their structural domains, the epiplasmins are defined as symmetric, asymmetric or atypical. The EST data aid in this classification, in the identification of putative regulating sequences such as TATA or CAAT boxes. When specific RNAi experiments were conducted using sequences from either symmetric or asymmetric classes, phenotypes were drastic. Local effects show either disrupted or ill-shaped epiplasmic scales. In either case, this results in aborted cell division. Using structural features, we show that 4 epiplasmins are also present in another ciliate, Tetrahymena thermophila. Their affiliation with the distinctive structural groups of Paramecium epiplasmins demonstrates an interspecific multigenic family. The epiplasmin multigenic family illustrates the history of genomic duplication in Paramecium. This study provides a framework which can guide functional analysis of epiplasmins, the major components of the membrane skeleton in ciliates. We show that this set of proteins handles an important developmental information in Paramecium since maintenance of epiplasm organization is crucial for cell morphogenesis.

  11. Revised systematics of Holospora-like bacteria and characterization of "Candidatus Gortzia infectiva", a novel macronuclear symbiont of Paramecium jenningsi.

    PubMed

    Boscaro, Vittorio; Fokin, Sergei I; Schrallhammer, Martina; Schweikert, Michael; Petroni, Giulio

    2013-01-01

    The genus Holospora (Rickettsiales) includes highly infectious nuclear symbionts of the ciliate Paramecium with unique morphology and life cycle. To date, nine species have been described, but a molecular characterization is lacking for most of them. In this study, we have characterized a novel Holospora-like bacterium (HLB) living in the macronuclei of a Paramecium jenningsi population. This bacterium was morphologically and ultrastructurally investigated in detail, and its life cycle and infection capabilities were described. We also obtained its 16S rRNA gene sequence and developed a specific probe for fluorescence in situ hybridization experiments. A new taxon, "Candidatus Gortzia infectiva", was established for this HLB according to its unique characteristics and the relatively low DNA sequence similarities shared with other bacteria. The phylogeny of the order Rickettsiales based on 16S rRNA gene sequences has been inferred, adding to the available data the sequence of the novel bacterium and those of two Holospora species (Holospora obtusa and Holospora undulata) characterized for the purpose. Our phylogenetic analysis provided molecular support for the monophyly of HLBs and showed a possible pattern of evolution for some of their features. We suggested to classify inside the family Holosporaceae only HLBs, excluding other more distantly related and phenotypically different Paramecium endosymbionts.

  12. Calcium signaling in closely related protozoan groups (Alveolata): non-parasitic ciliates (Paramecium, Tetrahymena) vs. parasitic Apicomplexa (Plasmodium, Toxoplasma).

    PubMed

    Plattner, H; Sehring, I M; Mohamed, I K; Miranda, K; De Souza, W; Billington, R; Genazzani, A; Ladenburger, E-M

    2012-05-01

    The importance of Ca2+-signaling for many subcellular processes is well established in higher eukaryotes, whereas information about protozoa is restricted. Recent genome analyses have stimulated such work also with Alveolates, such as ciliates (Paramecium, Tetrahymena) and their pathogenic close relatives, the Apicomplexa (Plasmodium, Toxoplasma). Here we compare Ca2+ signaling in the two closely related groups. Acidic Ca2+ stores have been characterized in detail in Apicomplexa, but hardly in ciliates. Two-pore channels engaged in Ca2+-release from acidic stores in higher eukaryotes have not been stingently characterized in either group. Both groups are endowed with plasma membrane- and endoplasmic reticulum-type Ca2+-ATPases (PMCA, SERCA), respectively. Only recently was it possible to identify in Paramecium a number of homologs of ryanodine and inositol 1,3,4-trisphosphate receptors (RyR, IP3R) and to localize them to widely different organelles participating in vesicle trafficking. For Apicomplexa, physiological experiments suggest the presence of related channels although their identity remains elusive. In Paramecium, IP3Rs are constitutively active in the contractile vacuole complex; RyR-related channels in alveolar sacs are activated during exocytosis stimulation, whereas in the parasites the homologous structure (inner membrane complex) may no longer function as a Ca2+ store. Scrutinized comparison of the two closely related protozoan phyla may stimulate further work and elucidate adaptation to parasitic life. See also "Conclusions" section. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Transposon Invasion of the Paramecium Germline Genome Countered by a Domesticated PiggyBac Transposase and the NHEJ Pathway

    PubMed Central

    Dubois, Emeline; Bischerour, Julien; Marmignon, Antoine; Mathy, Nathalie; Régnier, Vinciane; Bétermier, Mireille

    2012-01-01

    Sequences related to transposons constitute a large fraction of extant genomes, but insertions within coding sequences have generally not been tolerated during evolution. Thanks to their unique nuclear dimorphism and to their original mechanism of programmed DNA elimination from their somatic nucleus (macronucleus), ciliates are emerging model organisms for the study of the impact of transposable elements on genomes. The germline genome of the ciliate Paramecium, located in its micronucleus, contains thousands of short intervening sequences, the IESs, which interrupt 47% of genes. Recent data provided support to the hypothesis that an evolutionary link exists between Paramecium IESs and Tc1/mariner transposons. During development of the macronucleus, IESs are excised precisely thanks to the coordinated action of PiggyMac, a domesticated piggyBac transposase, and of the NHEJ double-strand break repair pathway. A PiggyMac homolog is also required for developmentally programmed DNA elimination in another ciliate, Tetrahymena. Here, we present an overview of the life cycle of these unicellular eukaryotes and of the developmentally programmed genome rearrangements that take place at each sexual cycle. We discuss how ancient domestication of a piggyBac transposase might have allowed Tc1/mariner elements to spread throughout the germline genome of Paramecium, without strong counterselection against insertion within genes. PMID:22888464

  14. Different polyamine pathways from bacteria have replaced eukaryotic spermidine biosynthesis in ciliates Tetrahymena thermophila and Paramecium tetaurelia.

    PubMed

    Li, Bin; Kim, Sok Ho; Zhang, Yang; Hanfrey, Colin C; Elliott, Katherine A; Ealick, Steven E; Michael, Anthony J

    2015-09-01

    The polyamine spermidine is absolutely required for growth and cell proliferation in eukaryotes, due to its role in post-translational modification of essential translation elongation factor eIF5A, mediated by deoxyhypusine synthase. We have found that free-living ciliates Tetrahymena and Paramecium lost the eukaryotic genes encoding spermidine biosynthesis: S-adenosylmethionine decarboxylase (AdoMetDC) and spermidine synthase (SpdSyn). In Tetrahymena, they were replaced by a gene encoding a fusion protein of bacterial AdoMetDC and SpdSyn, present as three copies. In Paramecium, a bacterial homospermidine synthase replaced the eukaryotic genes. Individual AdoMetDC-SpdSyn fusion protein paralogues from Tetrahymena exhibit undetectable AdoMetDC activity; however, when two paralogous fusion proteins are mixed, AdoMetDC activity is restored and spermidine is synthesized. Structural modelling indicates a functional active site is reconstituted by sharing critical residues from two defective protomers across the heteromer interface. Paramecium was found to accumulate homospermidine, suggesting it replaces spermidine for growth. To test this concept, a budding yeast spermidine auxotrophic strain was found to grow almost normally with homospermidine instead of spermidine. Biosynthesis of spermidine analogue aminopropylcadaverine, but not exogenously provided norspermidine, correlated with some growth. Finally, we found that diverse single-celled eukaryotic parasites and multicellular metazoan Schistosoma worms have lost the spermidine biosynthetic pathway but retain deoxyhypusine synthase. © 2015 John Wiley & Sons Ltd.

  15. The membrane skeleton in Paramecium: Molecular characterization of a novel epiplasmin family and preliminary GFP expression results.

    PubMed

    Pomel, Sébastien; Diogon, Marie; Bouchard, Philippe; Pradel, Lydie; Ravet, Viviane; Coffe, Gérard; Viguès, Bernard

    2006-02-01

    Previous attempts to identify the membrane skeleton of Paramecium cells have revealed a protein pattern that is both complex and specific. The most prominent structural elements, epiplasmic scales, are centered around ciliary units and are closely apposed to the cytoplasmic side of the inner alveolar membrane. We sought to characterize epiplasmic scale proteins (epiplasmins) at the molecular level. PCR approaches enabled the cloning and sequencing of two closely related genes by amplifications of sequences from a macronuclear genomic library. Using these two genes (EPI-1 and EPI-2), we have contributed to the annotation of the Paramecium tetraurelia macronuclear genome and identified 39 additional (paralogous) sequences. Two orthologous sequences were found in the Tetrahymena thermophila genome. Structural analysis of the 43 sequences indicates that the hallmark of this new multigenic family is a 79 aa domain flanked by two Q-, P- and V-rich stretches of sequence that are much more variable in amino-acid composition. Such features clearly distinguish members of the multigenic family from epiplasmic proteins previously sequenced in other ciliates. The expression of Green Fluorescent Protein (GFP)-tagged epiplasmin showed significant labeling of epiplasmic scales as well as oral structures. We expect that the GFP construct described herein will prove to be a useful tool for comparative subcellular localization of different putative epiplasmins in Paramecium.

  16. Trichocyst ribbons of a cryptomonads are constituted of homologs of R-body proteins produced by the intracellular parasitic bacterium of Paramecium.

    PubMed

    Yamagishi, Takahiro; Kai, Atsushi; Kawai, Hiroshi

    2012-04-01

    Trichocysts are ejectile organelles found in cryptomonads, dinoflagellates, and peniculine ciliates. The fine structure of trichocysts differs considerably among lineages, and their evolutionary relationships are unclear. The biochemical makeup of the trichocyst constituents has been studied in the ciliate Paramecium, but there have been no investigations of cryptomonads and dinoflagellates. Furthermore, morphological similarity between the contents of cryptomonad trichocysts and the R-bodies of the endosymbiotic bacteria of Paramecium has been reported. In this study, we identified the proteins of the trichocyst constituents in a red cryptomonad, Pyrenomonas helgolandii, and found their closest relationships to be with rebB that comprises the R-bodies of Caedibacter taeniospiralis (gammaproteobacteria), which is an endosymbiont of Paramecium. In addition, the biochemical makeups of the trichocysts are entirely different between cryptomonads and peniculine ciliates, and therefore, cryptomonad trichocysts have an evolutionary origin independent from the peniculine ciliate trichocysts.

  17. Sampling strategies for improving tree accuracy and phylogenetic analyses: a case study in ciliate protists, with notes on the genus Paramecium.

    PubMed

    Yi, Zhenzhen; Strüder-Kypke, Michaela; Hu, Xiaozhong; Lin, Xiaofeng; Song, Weibo

    2014-02-01

    In order to assess how dataset-selection for multi-gene analyses affects the accuracy of inferred phylogenetic trees in ciliates, we chose five genes and the genus Paramecium, one of the most widely used model protist genera, and compared tree topologies of the single- and multi-gene analyses. Our empirical study shows that: (1) Using multiple genes improves phylogenetic accuracy, even when their one-gene topologies are in conflict with each other. (2) The impact of missing data on phylogenetic accuracy is ambiguous: resolution power and topological similarity, but not number of represented taxa, are the most important criteria of a dataset for inclusion in concatenated analyses. (3) As an example, we tested the three classification models of the genus Paramecium with a multi-gene based approach, and only the monophyly of the subgenus Paramecium is supported. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. ISOLATION AND PARTIAL CHARACTERIZATION OF MACRO- AND MICRONUCLEI FROM PARAMECIUM AURELIA

    PubMed Central

    Cummings, Donald J.

    1972-01-01

    A method was developed for the isolation of macro- and micronuclei from Paramecium aurelia. This method utilized ionic and nonionic detergents to rupture the intact cells, calcium ions and spermidine were employed to protect the nuclei, and the nuclei were purified by centrifugation. Macronuclei consisted of 22% DNA, 10% RNA, and 68% protein. Micronuclei were composed of 9% DNA, 11% RNA, and 80% protein. DNA from both macro- and micronuclei had a density of 1.687 g/cc in CsCl and 1.417 g/cc in Cs2SO4. These values corresponded to G + C content of about 23%. The RNA of macronuclei was examined by gel electrophoresis, and two high molecular weight species were identified having molecular Weights of 1.3 x 106 and 2.8 x 106 daltons. Three syngens were studied, and in each case the conditions for isolation of the nuclei were the same and no differences were observed in the properties of the nuclei. PMID:4552140

  19. Photoreactivation in Paramecium tetraurelia under conditions of various degrees of ozone layer depletion.

    PubMed

    Takahashi, Akihisa; Kumatani, Toshihiro; Usui, Saori; Tsujimura, Ryoko; Seki, Takaharu; Morimoto, Kouichi; Ohnishi, Takeo

    2005-01-01

    Photoreactivation (PR) is an efficient survival mechanism that helps protect cells against the harmful effects of solar-ultraviolet (UV) radiation. The PR mechanism involves photolyase, just one enzyme, and can repair DNA damage, such as cyclobutane-pyrimidine dimers (CPD) induced by near-UV/blue light, a component of sunlight. Although the balance of near-UV/blue light and far-UV light reaching the Earth's surface could be altered by the atmospheric ozone layer's depletion, experiments simulating this environmental change and its possible effects on life have not yet been performed. To quantify the strength of UVB in sunlight reaching the Earth's surface, we measured the number of CPD generated in plasmid DNA after UVB irradiation or exposure to sunlight. To simulate the increase of solar-UV radiation resulting from the ozone layer depletion, Paramecium tetraurelia was exposed to UVB and/or sunlight in clear summer weather. PR recovery after exposure to sunlight was complete at a low dose rate of 0.2 J/m2 x s, but was less efficient when the dose rate was increased by a factor of 2.5 to 0.5 J/m2 x s. It is suggested that solar-UV radiation would not influence the cell growth of P. tetraurelia for the reason of high PR activity even when the ozone concentration was decreased 30% from the present levels.

  20. Ionic Control of the Reversal Response of Cilia in Paramecium caudatum

    PubMed Central

    Naitoh, Yutaka

    1968-01-01

    The duration of ciliary reversal of Paramecium caudatum in response to changes in external ionic factors was determined with various ionic compositions of both equilibration and stimulation media. The reversal response was found to occur when calcium ions bound by an inferred cellular cation exchange system were liberated in exchange for externally applied cations other than calcium. Factors which affect the duration of the response were (a) initial amount of calcium bound by the cation exchange system, (b) final amount of calcium bound by the system after equilibration with the stimulation medium, and (c) concentration of calcium ions in the stimulation medium. An empirical equation is presented which relates the duration of the response to these three factors. On the basis of these and previously published data, the following hypothesis is proposed for the mechanism underlying ciliary reversal in response to cationic stimulation: Ca++ liberated from the cellular cation exchange system activates a contractile system which is energized by ATP. Contraction of this component results in the reversal of effective beat direction of cilia by a mechanism not yet understood. The duration of reversal in live paramecia is related to the time course of bound calcium release. PMID:4966766

  1. Side-effects of fenazaquin on a cellular model of Paramecium.

    PubMed

    Benbouzid, Houneïda; Berrebbah, Houria; Djebar, Mohammed-Réda; Smagghe, Guy

    2009-01-01

    Our biodiversity has long been preserved, but the main constituents of our environment have been particularly affected by the addition of molecules resulting from agricultural and industrial activities. It is well accepted that these changes may stress some species, making them more vulnerable. In this project, we determined the disruptive side-effects of a pesticide on several biochemical endpoints and the behaviour of a microorganism as the ciliate protist Paramecium sp. Here we used fenazaquin [4-(4-tert-butylphenethoxy)quinazoline] that belongs to the quinazoline class of chemicals and that is a pesticide intended to control mites and insects; its route of exposure is ingestion and dermal, and its mode of action is the disruption of the biochemistry of insect mitochondria. In our experiments with fenazaquin at 40, 60 and 80 nM, we recorded disturbances in protein and glutathione, in glutathione S-transferase, and a decrease in consumption of oxygen. The results are discussed in relation to potential risks and mechanisms of action. In addition, the data can be used as reference values in further testing with other pesticides and chemistries.

  2. Spliced DNA Sequences in the Paramecium Germline: Their Properties and Evolutionary Potential

    PubMed Central

    Catania, Francesco; McGrath, Casey L.; Doak, Thomas G.; Lynch, Michael

    2013-01-01

    Despite playing a crucial role in germline-soma differentiation, the evolutionary significance of developmentally regulated genome rearrangements (DRGRs) has received scant attention. An example of DRGR is DNA splicing, a process that removes segments of DNA interrupting genic and/or intergenic sequences. Perhaps, best known for shaping immune-system genes in vertebrates, DNA splicing plays a central role in the life of ciliated protozoa, where thousands of germline DNA segments are eliminated after sexual reproduction to regenerate a functional somatic genome. Here, we identify and chronicle the properties of 5,286 sequences that putatively undergo DNA splicing (i.e., internal eliminated sequences [IESs]) across the genomes of three closely related species of the ciliate Paramecium (P. tetraurelia, P. biaurelia, and P. sexaurelia). The study reveals that these putative IESs share several physical characteristics. Although our results are consistent with excision events being largely conserved between species, episodes of differential IES retention/excision occur, may have a recent origin, and frequently involve coding regions. Our findings indicate interconversion between somatic—often coding—DNA sequences and noncoding IESs, and provide insights into the role of DNA splicing in creating potentially functional genetic innovation. PMID:23737328

  3. Epiplasmins and epiplasm in paramecium: the building of a submembraneous cytoskeleton.

    PubMed

    Aubusson-Fleury, Anne; Bricheux, Geneviève; Damaj, Raghida; Lemullois, Michel; Coffe, Gérard; Donnadieu, Florence; Koll, France; Viguès, Bernard; Bouchard, Philippe

    2013-07-01

    In ciliates, basal bodies and associated appendages are bound to a submembrane cytoskeleton. In Paramecium, this cytoskeleton takes the form of a thin dense layer, the epiplasm, segmented into regular territories, the units where basal bodies are inserted. Epiplasmins, the main component of the epiplasm, constitute a large family of 51 proteins distributed in 5 phylogenetic groups, each characterized by a specific molecular design. By GFP-tagging, we analyzed their differential localisation and role in epiplasm building and demonstrated that: 1) The epiplasmins display a low turnover, in agreement with the maintenance of an epiplasm layer throughout the cell cycle; 2) Regionalisation of proteins from different groups allows us to define rim, core, ring and basal body epiplasmins in the interphase cell; 3) Their dynamics allows definition of early and late epiplasmins, detected early versus late in the duplication process of the units. Epiplasmins from each group exhibit a specific combination of properties. Core and rim epiplasmins are required to build a unit; ring and basal body epiplasmins seem more dispensable, suggesting that they are not required for basal body docking. We propose a model of epiplasm unit assembly highlighting its implication in structural heredity in agreement with the evolutionary history of epiplasmins. Copyright © 2013 Elsevier GmbH. All rights reserved.

  4. The Evolutionary Relationships between Endosymbiotic Green Algae of Paramecium bursaria Syngens Originating from Different Geographical Locations.

    PubMed

    Zagata, Patrycja; Greczek-Stachura, Magdalena; Tarcz, Sebastian; Rautian, Maria

    2016-01-01

    Paramecium bursaria (Ehrenberg 1831), a freshwater ciliate, typically harbors hundreds of green algal symbionts inside the cell. The aim of present study was the molecular identification of newly analyzed P. bursaria symbionts. The second aspect of the present survey was testing a hypothesis whether endosymbionts prefer the specified syngen of the host, and the specified geographical distribution. Ten strains of endosymbionts isolated from strains of P. bursaria originating from different geographical locations were studied. We analyzed for the first time, both the fragment of plastid genome containing 3'rpl36-5' infA genes and a fragment of a nuclear gene encoding large subunit ribosomal RNA (LSU rDNA). The analysis of the LSU rDNA sequences showed the existence of 3 haplotypes and the haplotype diversity of 0.733, and 8 haplotypes for the 3'rpl36-5' infA gene fragment and haplotype diversity of 0.956. The endosymbionts isolated from P. bursaria strains were identified as Chlorella vulgaris, Ch. variabilis and Micractinium conductrix. There was no correlation between the syngen of P. bursaria and the species of endosymbiont.

  5. Paramecium bursaria chlorella virus 1 proteome reveals novel architectural and regulatory features of a giant virus.

    PubMed

    Dunigan, David D; Cerny, Ronald L; Bauman, Andrew T; Roach, Jared C; Lane, Leslie C; Agarkova, Irina V; Wulser, Kurt; Yanai-Balser, Giane M; Gurnon, James R; Vitek, Jason C; Kronschnabel, Bernard J; Jeanniard, Adrien; Blanc, Guillaume; Upton, Chris; Duncan, Garry A; McClung, O William; Ma, Fangrui; Van Etten, James L

    2012-08-01

    The 331-kbp chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) genome was resequenced and annotated to correct errors in the original 15-year-old sequence; 40 codons was considered the minimum protein size of an open reading frame. PBCV-1 has 416 predicted protein-encoding sequences and 11 tRNAs. A proteome analysis was also conducted on highly purified PBCV-1 virions using two mass spectrometry-based protocols. The mass spectrometry-derived data were compared to PBCV-1 and its host Chlorella variabilis NC64A predicted proteomes. Combined, these analyses revealed 148 unique virus-encoded proteins associated with the virion (about 35% of the coding capacity of the virus) and 1 host protein. Some of these proteins appear to be structural/architectural, whereas others have enzymatic, chromatin modification, and signal transduction functions. Most (106) of the proteins have no known function or homologs in the existing gene databases except as orthologs with proteins of other chloroviruses, phycodnaviruses, and nuclear-cytoplasmic large DNA viruses. The genes encoding these proteins are dispersed throughout the virus genome, and most are transcribed late or early-late in the infection cycle, which is consistent with virion morphogenesis.

  6. On the nature of species: insights from Paramecium and other ciliates

    PubMed Central

    Hall, Meaghan S.; Katz, Laura A.

    2011-01-01

    The multiple species concepts currently in use by the scientific community (e.g. Morphological, Biological, Phylogenetic) are united in that they all aim to capture the process of divergence between populations. For example, the Biological Species Concept (BSC) defines a species as a natural group of organisms that is reproductively isolated from other such groups. Here we synthesize nearly a century of research on the ciliate genus Paramecium that highlights the shortcomings of our prevailing notions on the nature of species. In this lineage, there is discordance between morphology, mating behavior, and genetics, features assumed to be correlated, at least after sufficient time has passed, under all species concepts. Intriguingly, epigenetic phenomena are well documented in ciliates where they influence features such as germline/soma differentiation and mating type determination. Consequently, we hypothesize that divergence within ciliate populations is due to a dynamic interaction between genetic and epigenetic factors. The growing list of examples of epigenetic phenomena that potentially impact speciation (i.e. by influencing the dynamics of sex chromosomes, fate of hybrids, zygotic drive and genomic conflicts) suggests that interactions between genetics and epigenetics may also drive divergence in other eukaryotic lineages. PMID:21505762

  7. Sas-4 proteins are required during basal body duplication in Paramecium

    PubMed Central

    Gogendeau, Delphine; Hurbain, Ilse; Raposo, Graca; Cohen, Jean; Koll, France; Basto, Renata

    2011-01-01

    Centrioles and basal bodies are structurally related organelles composed of nine microtubule (MT) triplets. Studies performed in Caenorhabditis elegans embryos have shown that centriole duplication takes place in sequential way, in which different proteins are recruited in a specific order to assemble a procentriole. ZYG-1 initiates centriole duplication by triggering the recruitment of a complex of SAS-5 and SAS-6, which then recruits the final player, SAS-4, to allow the incorporation of MT singlets. It is thought that a similar mechanism (that also involves additional proteins) is present in other animal cells, but it remains to be investigated whether the same players and their ascribed functions are conserved during basal body duplication in cells that exclusively contain basal bodies. To investigate this question, we have used the multiciliated protist Paramecium tetraurelia. Here we show that in the absence of PtSas4, two types of defects in basal body duplication can be identified. In the majority of cases, the germinative disk and cartwheel, the first structures assembled during duplication, are not detected. In addition, if daughter basal bodies were formed, they invariably had defects in MT recruitment. Our results suggest that PtSas4 has a broader function than its animal orthologues. PMID:21289083

  8. Virus-host interactions: insights from the replication cycle of the large Paramecium bursaria chlorella virus.

    PubMed

    Milrot, Elad; Mutsafi, Yael; Fridmann-Sirkis, Yael; Shimoni, Eyal; Rechav, Katya; Gurnon, James R; Van Etten, James L; Minsky, Abraham

    2016-01-01

    The increasing interest in cytoplasmic factories generated by eukaryotic-infecting viruses stems from the realization that these highly ordered assemblies may contribute fundamental novel insights to the functional significance of order in cellular biology. Here, we report the formation process and structural features of the cytoplasmic factories of the large dsDNA virus Paramecium bursaria chlorella virus 1 (PBCV-1). By combining diverse imaging techniques, including scanning transmission electron microscopy tomography and focused ion beam technologies, we show that the architecture and mode of formation of PBCV-1 factories are significantly different from those generated by their evolutionary relatives Vaccinia and Mimivirus. Specifically, PBCV-1 factories consist of a network of single membrane bilayers acting as capsid templates in the central region, and viral genomes spread throughout the host cytoplasm but excluded from the membrane-containing sites. In sharp contrast, factories generated by Mimivirus have viral genomes in their core, with membrane biogenesis region located at their periphery. Yet, all viral factories appear to share structural features that are essential for their function. In addition, our studies support the notion that PBCV-1 infection, which was recently reported to result in significant pathological outcomes in humans and mice, proceeds through a bacteriophage-like infection pathway. © 2015 John Wiley & Sons Ltd.

  9. Characteristics of the digestive vacuole membrane of the alga-bearing ciliate Paramecium bursaria.

    PubMed

    Kodama, Yuuki; Fujishima, Masahiro

    2012-07-01

    Cells of the ciliate Paramecium bursaria harbor symbiotic Chlorella spp. in their cytoplasm. To establish endosymbiosis with alga-free P. bursaria, symbiotic algae must leave the digestive vacuole (DV) to appear in the cytoplasm by budding of the DV membrane. This budding was induced not only by intact algae but also by boiled or fixed algae. However, this budding was not induced when food bacteria or India ink were ingested into the DVs. These results raise the possibility that P. bursaria can recognize sizes of the contents in the DVs. To elucidate this possibility, microbeads with various diameters were mixed with alga-free P. bursaria and traced their fate. Microbeads with 0.20μm diameter did not induce budding of the DVs. Microbeads with 0.80μm diameter produced DVs of 5-10μm diameter at 3min after mixing; then the DVs fragmented and became vacuoles of 2-5μm diameter until 3h after mixing. Each microbead with a diameter larger than 3.00μm induced budding similarly to symbiotic Chlorella. These observations reveal that induction of DV budding depends on the size of the contents in the DVs. Dynasore, a dynamin inhibitor, greatly inhibited DV budding, suggesting that dynamin might be involved in DV budding. Copyright © 2011 Elsevier GmbH. All rights reserved.

  10. Spliced DNA sequences in the Paramecium germline: their properties and evolutionary potential.

    PubMed

    Catania, Francesco; McGrath, Casey L; Doak, Thomas G; Lynch, Michael

    2013-01-01

    Despite playing a crucial role in germline-soma differentiation, the evolutionary significance of developmentally regulated genome rearrangements (DRGRs) has received scant attention. An example of DRGR is DNA splicing, a process that removes segments of DNA interrupting genic and/or intergenic sequences. Perhaps, best known for shaping immune-system genes in vertebrates, DNA splicing plays a central role in the life of ciliated protozoa, where thousands of germline DNA segments are eliminated after sexual reproduction to regenerate a functional somatic genome. Here, we identify and chronicle the properties of 5,286 sequences that putatively undergo DNA splicing (i.e., internal eliminated sequences [IESs]) across the genomes of three closely related species of the ciliate Paramecium (P. tetraurelia, P. biaurelia, and P. sexaurelia). The study reveals that these putative IESs share several physical characteristics. Although our results are consistent with excision events being largely conserved between species, episodes of differential IES retention/excision occur, may have a recent origin, and frequently involve coding regions. Our findings indicate interconversion between somatic--often coding--DNA sequences and noncoding IESs, and provide insights into the role of DNA splicing in creating potentially functional genetic innovation.

  11. A Ca sup 2+ influx associated with exocytosis is specifically abolished in a Paramecium exocytotic mutant

    SciTech Connect

    Kerboeuf, D.; Cohen, J. )

    1990-12-01

    A Paramecium possesses secretory organelles called trichocysts which are docked beneath the plasma membrane awaiting an external stimulus that triggers their exocytosis. Membrane fusion is the sole event provoked by the stimulation and can therefore be studied per se. Using 3 microM aminoethyl dextran as a vital secretagogue, we analyzed the movements of calcium (Ca{sup 2+}) during the discharge of trichocysts. We showed that (a) external Ca{sup 2+}, at least at 3 X 10(-7) M, is necessary for AED to induce exocytosis; (b) a dramatic and transient influx of Ca{sup 2+} as measured from {sup 45}Ca uptake is induced by AED; (c) this influx is independent of the well-characterized voltage-operated Ca{sup 2+} channels of the ciliary membranes since it persists in a mutant devoid of these channels; and (d) this influx is specifically abolished in one of the mutants unable to undergo exocytosis, nd12. We propose that the Ca{sup 2+} influx induced by AED reflects an increase in membrane permeability through the opening of novel Ca{sup 2+} channel or the activation of other Ca{sup 2+} transport mechanism in the plasma membrane. The resulting rise in cytosolic Ca{sup 2+} concentration would in turn induce membrane fusion. The mutation nd12 would affect a gene product involved in the control of plasma membrane permeability to Ca{sup 2+}, specifically related to membrane fusion.

  12. Timing of Developmentally Programmed Excision and Circularization of Paramecium Internal Eliminated Sequences

    PubMed Central

    Bétermier, Mireille; Duharcourt, Sandra; Seitz, Hervé; Meyer, Eric

    2000-01-01

    Paramecium internal eliminated sequences (IESs) are short AT-rich DNA elements that are precisely eliminated from the germ line genome during development of the somatic macronucleus. They are flanked by one 5′-TA-3′ dinucleotide on each side, a single copy of which remains at the donor site after excision. The timing of their excision was examined in synchronized conjugating cells by quantitative PCR. Significant amplification of the germ line genome was observed prior to IES excision, which starts 12 to 14 h after initiation of conjugation and extends over a 2- to 4-h period. Following excision, two IESs were shown to form extrachromosomal circles that can be readily detected on Southern blots of genomic DNA from cells undergoing macronuclear development. On these circular molecules, covalently joined IES ends are separated by one copy of the flanking 5′-TA-3′ repeat. The similar structures of the junctions formed on the excised and donor molecules point to a central role for this dinucleotide in IES excision. PMID:10669733

  13. Maintenance of algal endosymbionts in Paramecium bursaria: a simple model based on population dynamics.

    PubMed

    Iwai, Sosuke; Fujiwara, Kenji; Tamura, Takuro

    2016-09-01

    Algal endosymbiosis is widely distributed in eukaryotes including many protists and metazoans, and plays important roles in aquatic ecosystems, combining phagotrophy and phototrophy. To maintain a stable symbiotic relationship, endosymbiont population size in the host must be properly regulated and maintained at a constant level; however, the mechanisms underlying the maintenance of algal endosymbionts are still largely unknown. Here we investigate the population dynamics of the unicellular ciliate Paramecium bursaria and its Chlorella-like algal endosymbiont under various experimental conditions in a simple culture system. Our results suggest that endosymbiont population size in P. bursaria was not regulated by active processes such as cell division coupling between the two organisms, or partitioning of the endosymbionts at host cell division. Regardless, endosymbiont population size was eventually adjusted to a nearly constant level once cells were grown with light and nutrients. To explain this apparent regulation of population size, we propose a simple mechanism based on the different growth properties (specifically the nutrient requirements) of the two organisms, and based from this develop a mathematical model to describe the population dynamics of host and endosymbiont. The proposed mechanism and model may provide a basis for understanding the maintenance of algal endosymbionts. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  14. Microtubules mediate germ-nuclear behavior after meiosis in conjugation of Paramecium caudatum.

    PubMed

    Nakajima, Yuka; Ishida, Masaki; Mikami, Kazuyuki

    2002-01-01

    Microtubule dynamics in Paramecium caudatum were investigated with an anti-alpha-tubulin antibody and a microinjection technique to determine the function of microtubules on micronuclear behavior during conjugation. After meiosis, all four haploid micronuclei were connected by microtubular filaments to the paroral region and moved close to this region. This nuclear movement was micronucleus-specific, because some small macronuclear fragments transplanted from exconjugants never moved to the region. Only one of the four germ nuclei moved into the paroral cone and was covered by microtubule assembly (the so-called first assembly of microtubules, AM-I). This nucleus survived there, while the other three not in this region degenerated. The movement of germ nucleus was inhibited by the injection of the anti-alpha-tubulin antibody. The surviving germ nucleus divided once and produced a migratory pronucleus and a stationary pronucleus. Prior to the reciprocal exchange of the migratory nuclei, microtubules assembled around the migratory pronuclei again (the so-called second assembly of microtubules, AM-II). Then, the migratory pronucleus moved into the partner cell and fused with the stationary pronucleus. Thus, microtubules appear to be indispensable for nuclear behavior: they enable migration of postmeiotic nuclei to the paroral region and they permit the survival of the nucleus at the paroral cone.

  15. Variability of autogamy-maturation pattern in genetically identical populations of Paramecium tetraurelia.

    PubMed

    Komori, Rie; Harumoto, Terue; Fujisawa, Hiromi; Takagi, Yoshiomi

    2002-11-01

    Autogamy in Paramecium tetraurelia is a form of sexual reproduction in a single cell that results in homozygosity in every genetic locus. Autogamy becomes inducible by natural starvation several fissions after the previous autogamy, and percent autogamy increases gradually with clonal age to reach 100%. We here report the degree of variability of the autogamy-maturation pattern, and how it is inherited through autogamous generations. We assessed the autogamy-maturation pattern by monitoring percent autogamy at the ages of 9, 18 and 27 fissions in the wild-type stock 51. To determine how the autogamy-maturation pattern is inherited, clones that showed the lowest and the highest percent autogamy at age 18 in a given autogamous generation (Gn) were examined for their percent autogamy in the next autogamous generation (Gn+1). This procedure was repeated through successive autogamous generations. We found that percent autogamy at ages 9 and 27 was rather stable (low and high, respectively), while it was extremely variable at age 18 ranging from 3% to 100%. We also found that percent autogamy at age 18 in the progeny clones was variable irrespective of percent autogamy at age 18 in the parental clones; there was no regular rule such as producing progeny with higher (or lower) percent autogamy from parents with lower (or higher) percent autogamy.

  16. The molecular basis for the alternative stable phenotype in a behavioral mutant of Paramecium tetraurelia.

    PubMed

    Matsuda, A; Takahashi, M

    2001-10-01

    In the sexual reproduction of Paramecium tetraurelia, the somatic nucleus (macronucleus) undergoes massive genomic rearrangement, including gene amplification and excision of internal eliminated sequences (IESs), in its normal developmental process. Strain d4-662, one of the pawn mutants, is a behavioral mutant of P. tetraurelia that carries a recessive allele of pwB662. ThepwB gene in the macronucleus of the strain has an insertion of the IES because a base substitution within the IES prevents its excision during gene rearrangement. Cultures of this strain frequently contain cells reverting to the wild type in the behavioral phenotype. The mutant and revertant cells maintained stable clonal phenotypes under the various environmental conditions examined unless they underwent sexual reproduction. After sexual reproduction, both mutant and revertant produced 2.7-7.1% reverted progeny. A molecular analysis performed on the macronuclear DNA of the mutant and revertant of d4-662 showed that much less than 1% of the mutant IES was precisely excised at every sexual reproduction of the strain. Therefore, the alternative phenotype of strain d4-662 seems to be caused by an alternative excision of the mutant IES.

  17. Reverse evolution: selection against costly resistance in disease-free microcosm populations of Paramecium caudatum.

    PubMed

    Duncan, Alison B; Fellous, Simon; Kaltz, Oliver

    2011-12-01

    Evolutionary costs of parasite resistance arise if genes conferring resistance reduce fitness in the absence of parasites. Thus, parasite-mediated selection may lead to increased resistance and a correlated decrease in fitness, whereas relaxed parasite-mediated selection may lead to reverse evolution of increased fitness and a correlated decrease in resistance. We tested this idea in experimental populations of the protozoan Paramecium caudatum and the parasitic bacterium Holospora undulata. After eight years, resistance to infection and asexual reproduction were compared among paramecia from (1) "infected" populations, (2) uninfected "naive" populations, and (3) previously infected, parasite-free "recovered" populations. Paramecia from "infected" populations were more resistant (+12%), but had lower reproduction (-15%) than "naive" paramecia, indicating an evolutionary trade-off between resistance and fitness. Recovered populations showed similar reproduction to naive populations; however, resistance of recently (<3 years) recovered populations was similar to paramecia from infected populations, whereas longer (>3 years) recovered populations were as susceptible as naive populations. This suggests a weak, convex trade-off between resistance and fitness, allowing recovery of fitness, without complete loss of resistance, favoring the maintenance of a generalist strategy of intermediate fitness and resistance. Our results indicate that (co)evolution with parasites can leave a genetic signature in disease-free populations. © 2011 The Author(s). Evolution© 2011 The Society for the Study of Evolution.

  18. A Suppressor Gene Involved in Chemical Induction of Conjugation in PARAMECIUM AURELIA

    PubMed Central

    Cronkite, Donald L.

    1975-01-01

    A dominant gene Su(kau–2) partially suppresses the effect of an apparently unlinked recessive kau–2. Gene kau–2 blocks chemical induction of conjugation in Paramecium aurelia syngen 8 when solutions of acriflavine + either KCl or MgCl 2 are used. Wild-type cells are induced to conjugate in either solution. When cells homozygous for kau–2 also have gene Su(kau–2) , they are still uninducible in the solution containing KCl, but become inducible in the MgCl2 solution. Analysis of the concentrations of solutions which are effective in induction of conjugation of various genotypes shows that the action of Su(kau–2) is not a simple restoration of wild-type phenotype since certain novel features of the suppressor can be seen. Analysis of the duration of ciliary reversal of various genotypes suggests that one necessary step in chemical induction of conjugation is a certain magnitude of deplorization of the surface membrane of the cell. PMID:1137967

  19. Epigenetic self-regulation of developmental excision of an internal eliminated sequence on Paramecium tetraurelia.

    PubMed

    Duharcourt, S; Butler, A; Meyer, E

    1995-08-15

    Differentiation of the somatic macronucleus of ciliates after sexual events involves the programmed excision of thousands of single-copy internal eliminated sequences (IESs) from the germ-line genome. We have studied two cell lines of Paramecium tetraurelia that have identical germ-line genomes but differ in their macronuclear genomes. In the IES- cell line, a 222-bp IES interrupting a coding sequence is reproducibly excised during macronuclear differentiation, whereas it is not in the IES+ cell line. In a cross between the two lines, the developmental alternative in maternally inherited, suggesting that it is epigenetically controlled by the old (prezygotic) macronucleus in each cell. Transformation of the macronucleus of both lines with plasmids carrying fragments of either version of the gene shows that the presence of the IES sequence in the old macronucleus results in retention of the IES in the new macronuclear genome of sexual progeny. This could be attributable to (1) inhibition of excision, or (2) repair of a double-strand gap left in the genomic sequence after constitutive excision of the IES, by a polymerization mechanism using a homologous IES+ template from the old macronucleus. The latter possibility is ruled out by experiments showing that modified IESs can inhibit excision without being copied in the new macronuclear genome. Possible mechanisms are discussed in the light of a quantitative analysis of excision inhibition by the maternal IES sequence.

  20. Silencing-associated and meiosis-specific small RNA pathways in Paramecium tetraurelia.

    PubMed

    Lepère, Gersende; Nowacki, Mariusz; Serrano, Vincent; Gout, Jean-François; Guglielmi, Gérard; Duharcourt, Sandra; Meyer, Eric

    2009-02-01

    Distinct small RNA pathways are involved in the two types of homology-dependent effects described in Paramecium tetraurelia, as shown by a functional analysis of Dicer and Dicer-like genes and by the sequencing of small RNAs. The siRNAs that mediate post-transcriptional gene silencing when cells are fed with double-stranded RNA (dsRNA) were found to comprise two subclasses. DCR1-dependent cleavage of the inducing dsRNA generates approximately 23-nt primary siRNAs from both strands, while a different subclass of approximately 24-nt RNAs, characterized by a short untemplated poly-A tail, is strictly antisense to the targeted mRNA, suggestive of secondary siRNAs that depend on an RNA-dependent RNA polymerase. An entirely distinct pathway is responsible for homology-dependent regulation of developmental genome rearrangements after sexual reproduction. During early meiosis, the DCL2 and DCL3 genes are required for the production of a highly complex population of approximately 25-nt scnRNAs from all types of germline sequences, including both strands of exons, introns, intergenic regions, transposons and Internal Eliminated Sequences. A prominent 5'-UNG signature, and a minor fraction showing the complementary signature at positions 21-23, indicate that scnRNAs are cleaved from dsRNA precursors as duplexes with 2-nt 3' overhangs at both ends, followed by preferential stabilization of the 5'-UNG strand.

  1. Non-Mendelian inheritance induced by gene amplification in the germ nucleus of Paramecium tetraurelia.

    PubMed

    Matsuda, Atsushi; Takahashi, Mihoko

    2005-01-01

    A genetic investigation of strain d4-95, which carries a recessive mutant allele (pwB(95)) of pawn-B, one of the controlling elements of voltage-dependent calcium channels in Paramecium tetraurelia, revealed a non-Mendelian feature. Progeny of the cross between d4-95 and wild type often expressed a clonally stable mutant phenotype, even when they had a wild-type gene. The mutant phenotype was also expressed after self-fertilization of theoretical wild-type homozygotes recovered from the cross. Our molecular analysis demonstrated that the copy number of the mutant pwB gene in the micro- and macronucleus of d4-95 was much greater than that of the wild type. Most of the amplified, extra pwB gene copies in d4-95 were heritable independently from the original pwB locus. Repeated backcrossing of d4-95 with the wild type to dilute extra pwB genes in the strain produced segregants with a completely normal Mendelian trait in testcrosses. These results strongly suggest that a non-Mendelian inheritance of d4-95 was induced by gene amplification in the micronucleus.

  2. Vitamin E supplementation and intense selection increase clonal life span in Paramecium tetraurelia.

    PubMed

    Thomas, J; Nyberg, D

    1988-01-01

    Vitamin E added to standard Cerophyl medium at 0.025 mg/ml significantly increased the mean clonal life span of 32 lines of Paramecium tetraurelia from 52.5 days and 187 fissions to 59.9 days and 209 fissions (p less than .05) when compared to unsupplemented, paired controls. The age-specific death rates increased exponentially. Regression analyses found a significantly lower rate of increasing mortality for the supplemented lines compared to controls. Weekly fission rates of supplemented lines declined linearly; more slowly than controls, but not significantly so. A second experiment tested two higher levels of supplemental vitamin E (0.10 and 1.00 mg/ml). Sublines receiving 1.00 mg/ml of supplemental vitamin E had higher rates of mortality and lower fission rates initially, but the mortality rates increased and the fission rates decreased more slowly than in sublines receiving 0.10 mg/ml supplemental vitamin E, resulting in maximum clonal life spans of 141 days and 330 fissions (1.00 mg/ml sublines), compared to 74 days and 271 fissions (0.10 mg/ml sublines). Survivorship of the last individual cells (nondividing) of each clone followed an exponential decline, with a significant increase in mean survival time for supplemented compared to unsupplemented cells (p less than .05).

  3. Silencing-associated and meiosis-specific small RNA pathways in Paramecium tetraurelia

    PubMed Central

    Lepère, Gersende; Nowacki, Mariusz; Serrano, Vincent; Gout, Jean-François; Guglielmi, Gérard; Duharcourt, Sandra; Meyer, Eric

    2009-01-01

    Distinct small RNA pathways are involved in the two types of homology-dependent effects described in Paramecium tetraurelia, as shown by a functional analysis of Dicer and Dicer-like genes and by the sequencing of small RNAs. The siRNAs that mediate post-transcriptional gene silencing when cells are fed with double-stranded RNA (dsRNA) were found to comprise two subclasses. DCR1-dependent cleavage of the inducing dsRNA generates ∼23-nt primary siRNAs from both strands, while a different subclass of ∼24-nt RNAs, characterized by a short untemplated poly-A tail, is strictly antisense to the targeted mRNA, suggestive of secondary siRNAs that depend on an RNA-dependent RNA polymerase. An entirely distinct pathway is responsible for homology-dependent regulation of developmental genome rearrangements after sexual reproduction. During early meiosis, the DCL2 and DCL3 genes are required for the production of a highly complex population of ∼25-nt scnRNAs from all types of germline sequences, including both strands of exons, introns, intergenic regions, transposons and Internal Eliminated Sequences. A prominent 5′-UNG signature, and a minor fraction showing the complementary signature at positions 21–23, indicate that scnRNAs are cleaved from dsRNA precursors as duplexes with 2-nt 3′ overhangs at both ends, followed by preferential stabilization of the 5′-UNG strand. PMID:19103667

  4. Genetic evidence of "American" and "European" type symbiotic algae of Paramecium bursaria Ehrenberg.

    PubMed

    Hoshina, R; Kato, Y; Kamako, S; Imamura, N

    2005-09-01

    Paramecium bursaria is composed of a "host" ciliate and a "symbiont" green alga. Based upon physiology, DNA hybridization and virus infection, two types of symbionts, called "American" type and "European" type, have been reported to date. Here, we determined the 18S rDNA and internal transcribed spacer 2 (ITS2) regions for both "American" and "European" types. Sequence features clearly separated into two lineages; NC64A (USA), Syngen 2-3 (USA), Cs2 (Chinese), MRBG1 (Australian), and Japanese strains belong to the "American", whereas PB-SW1 (German) and CCAP 1660/11 (British) strains belong to the "European". In "American" 18S rDNA, three introns were inserted in the same positions as for previously described Japanese symbionts. In "European" 18S rDNA, a single intron occurred in a different position than in the "American". Between the types, sequence differences were seven or eight nucleotides (0.39 %) in the 18S rDNA exon, and more than 48 nucleotides (19.2 %) in ITS2 regions. We subsequently sequenced the host 18S rDNA. As a result, two groups: Cs2, MRBG1, and Japanese strains, and PB-SW1 and CCAP 1660/11 strains, were separated (with 23 substitutions and 4 insertions or deletions between the groups). The congruent separations between hosts and symbionts may imply that the type of symbiont depends on the host type.

  5. Cis-acting signals modulate the efficiency of programmed DNA elimination in Paramecium tetraurelia

    PubMed Central

    Ferro, Diana; Lepennetier, Gildas; Catania, Francesco

    2015-01-01

    In Paramecium, the regeneration of a functional somatic genome at each sexual event relies on the elimination of thousands of germline DNA sequences, known as Internal Eliminated Sequences (IESs), from the zygotic nuclear DNA. Here, we provide evidence that IESs’ length and sub-terminal bases jointly modulate IES excision by affecting DNA conformation in P. tetraurelia. Our study reveals an excess of complementary base pairing between IESs’ sub-terminal and contiguous sites, suggesting that IESs may form DNA loops prior to cleavage. The degree of complementary base pairing between IESs’ sub-terminal sites (termed Cin-score) is positively associated with IES length and is shaped by natural selection. Moreover, it escalates abruptly when IES length exceeds 45 nucleotides (nt), indicating that only sufficiently large IESs may form loops. Finally, we find that IESs smaller than 46 nt are favored targets of the cellular surveillance systems, presumably because of their relatively inefficient excision. Our findings extend the repertoire of cis-acting determinants for IES recognition/excision and provide unprecedented insights into the distinct selective pressures that operate on IESs and somatic DNA regions. This information potentially moves current models of IES evolution and of mechanisms of IES recognition/excision forward. PMID:26304543

  6. The native structure of cytoplasmic dynein at work translocating vesicles in Paramecium.

    PubMed

    Ishida, Masaki; Aihara, Marilynn S; Allen, Richard D; Fok, Agnes K

    2011-01-01

    In Paramecium multimicronucleatum, the discoidal vesicles, the acidosomes and the 100-nm carrier vesicles are involved in phagosome formation, phagosome acidification and endosomal processing, respectively. Numerous cross bridges link these vesicles to the kinetic side of the microtubules of a cytopharyngeal microtubular ribbon. Vesicles are translocated along these ribbons in a minus-end direction towards the cytopharynx. A monoclonal antibody specific for the light vanadate-photocleaved fragment of the heavy chain of cytoplasmic dynein was used to show that this dynein is located between the discoidal vesicles and the ribbons as well as on the cytosolic surface of the acidosomes and the 100-nm carrier vesicles. This antibody inhibited the docking of the vesicles to the microtubular ribbons so that the transport of discoidal vesicles and acidosomes were reduced by 60% and 70%, respectively. It had little effect on the dynein's velocity of translocation. These results show that cytoplasmic dynein is the motor for vesicle translocation and its location, between the vesicles and the ribbons, indicates that the cross bridges seen at this location in thin sections and in quick-frozen, deep-etched replicas are apparently the working dyneins. Such a working dynein cross bridge, as preserved by ultra-rapid freezing, is 54 nm long and has two legs arising from a globular head that appears to be firmly bound to its cargo vesicle and each leg consists of ≥3 beaded subunits with the last subunit making contact with the microtubular ribbon.

  7. Effect of Japanese Paramecium bursaria extract on photosynthetic carbon fixation of symbiotic algae.

    PubMed

    Kamako, Shin-ichiro; Imamura, Nobutaka

    2006-01-01

    To investigate the relationship between the Japanese Paramecium bursaria host and its symbiont, we studied the effect of a host cell-free extract on carbon fixation and photosynthate release of the symbiont. The host extract enhanced symbiotic algal carbon fixation about 3-fold at an increased concentration; however, release of photosynthate hardly changed. Since the enhancing effect was not affected by elimination of carbon dioxide from the host extract, the existence of a host factor that stimulates algal carbon fixation was made clear. The host factor is a heat-stable, low molecular weight substance. In relation to the pH dependence, the extract improved carbon fixation at acidic and neutral pH and showed almost no effect at pH 9.0. Therefore, the stimulation of carbon fixation by the host factor is unlikely to be caused by intracellular pH change. The extract also improved carbon fixation of several Chlorella species, symbiotic and free-living, and apparently exhibited no species specificity. Therefore, the host seems to regulate the photosynthesis of the symbiont via a specific compound.

  8. A Ca2+-activated ATPase specifically released by Ca2+ shock from Paramecium tetraurelia.

    PubMed

    Riddle, L M; Rauh, J J; Nelson, D L

    1982-06-14

    Deciliation of Paramecium tetraurelia by a Ca2+ shock procedure releases a discrete set of proteins which represent about 1% of the total cell protein. Marker enzymes for cytoplasm (hexokinase), endoplasmic reticulum (glucose-6-phosphatase), peroxisomes (catalase), and lysosomes (acid phosphatase) were not released by this treatment. Among the proteins selectively released is a Ca2+-dependent ATPase. This enzyme has a broad substrate specificity which includes GTP, ATP, and UTP, and it can be activated by Ca2+, Sr2+, or Ba2+, but not by Mg2+ or by monovalent cations. The crude enzyme has a specific activity of 2-3 mumol/min per mg; the optimal pH for activity is 7.5. ATPase, GTPase, and UTPase all reside in the same protein, which is inhibited by ruthenium red, is irreversibly denatured at 50 degrees C, and which has a sedimentation coefficient of 8-10 S. This enzyme is compared with other surface-derived ATPases of ciliated protozoans, and its possible roles are discussed.

  9. The Paramecium germline genome provides a niche for intragenic parasitic DNA: evolutionary dynamics of internal eliminated sequences.

    PubMed

    Arnaiz, Olivier; Mathy, Nathalie; Baudry, Céline; Malinsky, Sophie; Aury, Jean-Marc; Denby Wilkes, Cyril; Garnier, Olivier; Labadie, Karine; Lauderdale, Benjamin E; Le Mouël, Anne; Marmignon, Antoine; Nowacki, Mariusz; Poulain, Julie; Prajer, Malgorzata; Wincker, Patrick; Meyer, Eric; Duharcourt, Sandra; Duret, Laurent; Bétermier, Mireille; Sperling, Linda

    2012-01-01

    Insertions of parasitic DNA within coding sequences are usually deleterious and are generally counter-selected during evolution. Thanks to nuclear dimorphism, ciliates provide unique models to study the fate of such insertions. Their germline genome undergoes extensive rearrangements during development of a new somatic macronucleus from the germline micronucleus following sexual events. In Paramecium, these rearrangements include precise excision of unique-copy Internal Eliminated Sequences (IES) from the somatic DNA, requiring the activity of a domesticated piggyBac transposase, PiggyMac. We have sequenced Paramecium tetraurelia germline DNA, establishing a genome-wide catalogue of -45,000 IESs, in order to gain insight into their evolutionary origin and excision mechanism. We obtained direct evidence that PiggyMac is required for excision of all IESs. Homology with known P. tetraurelia Tc1/mariner transposons, described here, indicates that at least a fraction of IESs derive from these elements. Most IES insertions occurred before a recent whole-genome duplication that preceded diversification of the P. aurelia species complex, but IES invasion of the Paramecium genome appears to be an ongoing process. Once inserted, IESs decay rapidly by accumulation of deletions and point substitutions. Over 90% of the IESs are shorter than 150 bp and present a remarkable size distribution with a -10 bp periodicity, corresponding to the helical repeat of double-stranded DNA and suggesting DNA loop formation during assembly of a transpososome-like excision complex. IESs are equally frequent within and between coding sequences; however, excision is not 100% efficient and there is selective pressure against IES insertions, in particular within highly expressed genes. We discuss the possibility that ancient domestication of a piggyBac transposase favored subsequent propagation of transposons throughout the germline by allowing insertions in coding sequences, a fraction of the

  10. Multigene Family Encoding 3′,5′-Cyclic-GMP-Dependent Protein Kinases in Paramecium tetraurelia Cells

    PubMed Central

    Kissmehl, Roland; Krüger, Tim P.; Treptau, Tilman; Froissard, Marine; Plattner, Helmut

    2006-01-01

    In the ciliate Paramecium tetraurelia, 3′,5′-cyclic GMP (cGMP) is one of the second messengers involved in several signal transduction pathways. The enzymes for its production and degradation are well established for these cells, whereas less is known about the potential effector proteins. On the basis of a current Paramecium genome project, we have identified a multigene family with at least 35 members, all of which encode cGMP-dependent protein kinases (PKGs). They can be classified into 16 subfamilies with several members each. Two of the genes, PKG1-1 and PKG2-1, were analyzed in more detail after molecular cloning. They encode monomeric enzymes of 770 and 819 amino acids, respectively, whose overall domain organization resembles that in higher eukaryotes. The enzymes contain a regulatory domain of two tandem cyclic nucleotide-binding sites flanked by an amino-terminal region for intracellular localization and a catalytic domain with highly conserved regions for ATP binding and catalysis. However, some Paramecium PKGs show a different structure. In Western blots, PKGs are detected both as cytosolic and as structure-bound forms. Immunofluorescence labeling shows enrichment in the cell cortex, notably around the dense-core secretory vesicles (trichocysts), as well as in cilia. Immunogold electron microscopy analysis reveals consistent labeling of ciliary membranes, of the membrane complex composed of cell membrane and cortical Ca2+ stores, and of regions adjacent to ciliary basal bodies, trichocysts, and trafficking vesicles. Since PKGs (re)phosphorylate the exocytosis-sensitive phosphoprotein pp63/pf upon stimulation, the role of PKGs during stimulated exocytosis is discussed, in addition to a role in ciliary beat regulation. PMID:16400170

  11. Rare Freshwater Ciliate Paramecium chlorelligerum Kahl, 1935 and Its Macronuclear Symbiotic Bacterium “Candidatus Holospora parva”

    PubMed Central

    Lebedeva, Natalia; Migunova, Alexandra; Petroni, Giulio

    2016-01-01

    Ciliated protists often form symbioses with many diverse microorganisms. In particular, symbiotic associations between ciliates and green algae, as well as between ciliates and intracellular bacteria, are rather wide-spread in nature. In this study, we describe the complex symbiotic system between a very rare ciliate, Paramecium chlorelligerum, unicellular algae inhabiting its cytoplasm, and novel bacteria colonizing the host macronucleus. Paramecium chlorelligerum, previously found only twice in Germany, was retrieved from a novel location in vicinity of St. Petersburg in Russia. Species identification was based on both classical morphological methods and analysis of the small subunit rDNA. Numerous algae occupying the cytoplasm of this ciliate were identified with ultrastructural and molecular methods as representatives of the Meyerella genus, which before was not considered among symbiotic algae. In the same locality at least fifteen other species of “green” ciliates were found, thus it is indeed a biodiversity hot-spot for such protists. A novel species of bacterial symbionts living in the macronucleus of Paramecium chlorelligerum cells was morphologically and ultrastructurally investigated in detail with the description of its life cycle and infection capabilities. The new endosymbiont was molecularly characterized following the full-cycle rRNA approach. Furthermore, phylogenetic analysis confirmed that the novel bacterium is a member of Holospora genus branching basally but sharing all characteristics of the genus except inducing connecting piece formation during the infected host nucleus division. We propose the name “Candidatus Holospora parva” for this newly described species. The described complex system raises new questions on how these microorganisms evolve and interact in symbiosis. PMID:27992463

  12. The Paramecium Germline Genome Provides a Niche for Intragenic Parasitic DNA: Evolutionary Dynamics of Internal Eliminated Sequences

    PubMed Central

    Arnaiz, Olivier; Mathy, Nathalie; Baudry, Céline; Malinsky, Sophie; Aury, Jean-Marc; Denby Wilkes, Cyril; Garnier, Olivier; Labadie, Karine; Lauderdale, Benjamin E.; Le Mouël, Anne; Marmignon, Antoine; Nowacki, Mariusz; Poulain, Julie; Prajer, Malgorzata; Wincker, Patrick; Meyer, Eric; Duharcourt, Sandra; Duret, Laurent; Bétermier, Mireille; Sperling, Linda

    2012-01-01

    Insertions of parasitic DNA within coding sequences are usually deleterious and are generally counter-selected during evolution. Thanks to nuclear dimorphism, ciliates provide unique models to study the fate of such insertions. Their germline genome undergoes extensive rearrangements during development of a new somatic macronucleus from the germline micronucleus following sexual events. In Paramecium, these rearrangements include precise excision of unique-copy Internal Eliminated Sequences (IES) from the somatic DNA, requiring the activity of a domesticated piggyBac transposase, PiggyMac. We have sequenced Paramecium tetraurelia germline DNA, establishing a genome-wide catalogue of ∼45,000 IESs, in order to gain insight into their evolutionary origin and excision mechanism. We obtained direct evidence that PiggyMac is required for excision of all IESs. Homology with known P. tetraurelia Tc1/mariner transposons, described here, indicates that at least a fraction of IESs derive from these elements. Most IES insertions occurred before a recent whole-genome duplication that preceded diversification of the P. aurelia species complex, but IES invasion of the Paramecium genome appears to be an ongoing process. Once inserted, IESs decay rapidly by accumulation of deletions and point substitutions. Over 90% of the IESs are shorter than 150 bp and present a remarkable size distribution with a ∼10 bp periodicity, corresponding to the helical repeat of double-stranded DNA and suggesting DNA loop formation during assembly of a transpososome-like excision complex. IESs are equally frequent within and between coding sequences; however, excision is not 100% efficient and there is selective pressure against IES insertions, in particular within highly expressed genes. We discuss the possibility that ancient domestication of a piggyBac transposase favored subsequent propagation of transposons throughout the germline by allowing insertions in coding sequences, a fraction of the

  13. The conjugation-specific Die5 protein is required for development of the somatic nucleus in both Paramecium and Tetrahymena.

    PubMed

    Matsuda, Atsushi; Shieh, Annie Wan-Yi; Chalker, Douglas L; Forney, James D

    2010-07-01

    Development in ciliated protozoa involves extensive genome reorganization within differentiating macronuclei, which shapes the somatic genome of the next vegetative generation. Major events of macronuclear differentiation include excision of internal eliminated sequences (IESs), chromosome fragmentation, and genome amplification. Proteins required for these events include those with homology throughout eukaryotes as well as proteins apparently unique to ciliates. In this study, we identified the ciliate-specific Defective in IES Excision 5 (DIE5) genes of Paramecium tetraurelia (PtDIE5) and Tetrahymena thermophila (TtDIE5) as orthologs that encode nuclear proteins expressed exclusively during development. Abrogation of PtDie5 protein (PtDie5p) function by RNA interference (RNAi)-mediated silencing or TtDie5p by gene disruption resulted in the failure of developing macronuclei to differentiate into new somatic nuclei. Tetrahymena DeltaDIE5 cells arrested late in development and failed to complete genome amplification, whereas RNAi-treated Paramecium cells highly amplified new macronuclear DNA before the failure in differentiation, findings that highlight clear differences in the biology of these distantly related species. Nevertheless, IES excision and chromosome fragmentation failed to occur in either ciliate, which strongly supports that Die5p is a critical player in these processes. In Tetrahymena, loss of zygotic expression during development was sufficient to block nuclear differentiation. This observation, together with the finding that knockdown of Die5p in Paramecium still allows genome amplification, indicates that this protein acts late in macronuclear development. Even though DNA rearrangements in these two ciliates look to be quite distinct, analysis of DIE5 establishes the action of a conserved mechanism within the genome reorganization pathway.

  14. The Conjugation-Specific Die5 Protein Is Required for Development of the Somatic Nucleus in both Paramecium and Tetrahymena▿

    PubMed Central

    Matsuda, Atsushi; Shieh, Annie Wan-Yi; Chalker, Douglas L.; Forney, James D.

    2010-01-01

    Development in ciliated protozoa involves extensive genome reorganization within differentiating macronuclei, which shapes the somatic genome of the next vegetative generation. Major events of macronuclear differentiation include excision of internal eliminated sequences (IESs), chromosome fragmentation, and genome amplification. Proteins required for these events include those with homology throughout eukaryotes as well as proteins apparently unique to ciliates. In this study, we identified the ciliate-specific Defective in IES Excision 5 (DIE5) genes of Paramecium tetraurelia (PtDIE5) and Tetrahymena thermophila (TtDIE5) as orthologs that encode nuclear proteins expressed exclusively during development. Abrogation of PtDie5 protein (PtDie5p) function by RNA interference (RNAi)-mediated silencing or TtDie5p by gene disruption resulted in the failure of developing macronuclei to differentiate into new somatic nuclei. Tetrahymena ΔDIE5 cells arrested late in development and failed to complete genome amplification, whereas RNAi-treated Paramecium cells highly amplified new macronuclear DNA before the failure in differentiation, findings that highlight clear differences in the biology of these distantly related species. Nevertheless, IES excision and chromosome fragmentation failed to occur in either ciliate, which strongly supports that Die5p is a critical player in these processes. In Tetrahymena, loss of zygotic expression during development was sufficient to block nuclear differentiation. This observation, together with the finding that knockdown of Die5p in Paramecium still allows genome amplification, indicates that this protein acts late in macronuclear development. Even though DNA rearrangements in these two ciliates look to be quite distinct, analysis of DIE5 establishes the action of a conserved mechanism within the genome reorganization pathway. PMID:20495055

  15. Consensus inverted terminal repeat sequence of Paramecium IESs: resemblance to termini of Tc1-related and Euplotes Tec transposons.

    PubMed Central

    Klobutcher, L A; Herrick, G

    1995-01-01

    During the formation of a transcriptionally active macronucleus, ciliated protozoa excise large numbers of interstitial segments of DNA (internal eliminated sequences; IESs) from their chromosomes. In this study we analyze the published sequences of 20 IESs that interrupt surface protein genes of Paramecium and identify a consensus inverted terminal repeat. This sequence is similar to the ends of the Tc1-related transposons found in nematodes and other metazoans, as well as to both the ends of the Tec transposons and at least some of the IESs in the distantly related ciliate Euplotes crassus. The results of these analyses bolster previous proposals that IESs were created by transposition. PMID:7596830

  16. [The route of a bacterium Holospora in the cell of Paramecium (Ciliophora, Protista) from phagosome to the nucleus].

    PubMed

    Sabaneeva, E V; Fokin, S I; Kornilova, E S

    2002-01-01

    Problems encountered at the initial stages of stable symbiotic system formation are discussed in the review. The most studied models for interaction between pathogenic bacteria and metazoan cells are compared with a similar system including Paramecium (a ciliatte)--Holospora (a bacterium). Literary and our own data on the infection of P. caudatum with specific endocytobionts inhabiting the nuclear apparatus (H. obtusa in the macronucleus), and H. undulata (in the micronucleus) are analysed with respect to the modern understanding of the intracellular vesicle trafficking.

  17. Forced symbiosis between Synechocystis spp. PCC 6803 and apo-symbiotic Paramecium bursaria as an experimental model for evolutionary emergence of primitive photosynthetic eukaryotes.

    PubMed

    Ohkawa, Hiroshi; Hashimoto, Naoko; Furukawa, Shunsuke; Kadono, Takashi; Kawano, Tomonori

    2011-06-01

    Single-cell green paramecia (Paramecium bursaria) is a swimming vehicle that carries several hundred cells of endo-symbiotic green algae. Here, a novel model for endo-symbiosis, prepared by introducing and maintaining the cells of cyanobacterium (Synechocystis spp. PCC 6803) in the apo-symbiotic cells of P. bursaria is described.

  18. Characterization of multigene families in the micronuclear genome of Paramecium tetraurelia reveals a germline specific sequence in an intron of a centrin gene.

    PubMed Central

    Vayssié, L; Sperling, L; Madeddu, L

    1997-01-01

    In Paramecium, as in other ciliates, the transcriptionally active macronucleus is derived from the germline micronucleus by programmed DNA rearrangements, which include the precise excision of thousands of germline-specific sequences (internal eliminated sequences, IESs). We report the characterization of micronuclear versions of genes encoding Paramecium secretory granule proteins (trichocyst matrix proteins, TMPs) and Paramecium centrins. TMP and centrin multigene families, previously studied in the macronuclear genome, consist of genes that are co-expressed to provide mixtures of related polypeptides that co-assemble to form respectively the crystalline trichocyst matrix and the infraciliary lattice, a contractile cytoskeletal network. We present evidence that TMP and centrin genes identified in the macronucleus are also present in the micronucleus, ruling out the possibility that these novel multigene families are generated by somatic rearrangements during macronuclear development. No IESs were found in TMP genes, however, four IESs in or near germline centrin genes were characterized. The only intragenic IES is 75 bp in size, interrupts a 29 bp intron and is absent from at least one other closely related centrin gene. This is the first report of an IES in an intron in Paramecium. PMID:9023115

  19. Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum.

    PubMed

    Cui, Pengfei; Dong, Yuan; Li, Zhijian; Zhang, Yubo; Zhang, Shicui

    2016-07-01

    The global ever-growing concerns about multi-drug resistant (MDR) microbes leads to urgent demands for exploration of new antibiotics including antimicrobial peptides (AMPs). Here we demonstrated that a cDNA from Ciliata Paramecium caudatum, designated Pcamp1, coded for a protein with features characteristic of AMPs, which is not homologous to any AMPs currently known. Both the C-terminal 91 amino acid residues of PcAMP1, cPcAMP1, expressed in Escherichia coli and the C-terminal 26 amino acid residues (predicted mature AMP), cPcAMP1/26, synthesized, underwent a coil-to-helix transition in the presence of TFE, SDS or DPC. Functional assays revealed that cPcAMP1 and cPcAMP1/26 were both able to kill Aeromonas hydrophila and Staphylococcus aureus. ELISA showed that cPcAMP1 and cPcAMP1/26 were able to bind to microbe-associated molecular pattern molecules LPS and LTA, which was further corroborated by the observations that cPcAMP1 could deposit onto the bacterial membranes. Importantly, both cPcAMP1 and cPcAMP1/26 were able to induce bacterial membrane permeabilization and depolarization, and to increase intracellular ROS levels. Additionally, cPcAMP1 and cPcAMP1/26 were not cytotoxic to mammalian cells. Taken together, our results show that PcAMP1 is a potential AMP with a membrane selectivity towards bacterial cells, which renders it a promising template for the design of novel peptide antibiotics against MDR microbes. It also shows that use of signal conserved sequence of AMPs can be an effective tool to identify potential AMPs across different animal classes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Highly precise and developmentally programmed genome assembly in Paramecium requires ligase IV-dependent end joining.

    PubMed

    Kapusta, Aurélie; Matsuda, Atsushi; Marmignon, Antoine; Ku, Michael; Silve, Aude; Meyer, Eric; Forney, James D; Malinsky, Sophie; Bétermier, Mireille

    2011-04-01

    During the sexual cycle of the ciliate Paramecium, assembly of the somatic genome includes the precise excision of tens of thousands of short, non-coding germline sequences (Internal Eliminated Sequences or IESs), each one flanked by two TA dinucleotides. It has been reported previously that these genome rearrangements are initiated by the introduction of developmentally programmed DNA double-strand breaks (DSBs), which depend on the domesticated transposase PiggyMac. These DSBs all exhibit a characteristic geometry, with 4-base 5' overhangs centered on the conserved TA, and may readily align and undergo ligation with minimal processing. However, the molecular steps and actors involved in the final and precise assembly of somatic genes have remained unknown. We demonstrate here that Ligase IV and Xrcc4p, core components of the non-homologous end-joining pathway (NHEJ), are required both for the repair of IES excision sites and for the circularization of excised IESs. The transcription of LIG4 and XRCC4 is induced early during the sexual cycle and a Lig4p-GFP fusion protein accumulates in the developing somatic nucleus by the time IES excision takes place. RNAi-mediated silencing of either gene results in the persistence of free broken DNA ends, apparently protected against extensive resection. At the nucleotide level, controlled removal of the 5'-terminal nucleotide occurs normally in LIG4-silenced cells, while nucleotide addition to the 3' ends of the breaks is blocked, together with the final joining step, indicative of a coupling between NHEJ polymerase and ligase activities. Taken together, our data indicate that IES excision is a "cut-and-close" mechanism, which involves the introduction of initiating double-strand cleavages at both ends of each IES, followed by DSB repair via highly precise end joining. This work broadens our current view on how the cellular NHEJ pathway has cooperated with domesticated transposases for the emergence of new mechanisms

  1. Functional study of genes essential for autogamy and nuclear reorganization in Paramecium.

    PubMed

    Nowak, Jacek K; Gromadka, Robert; Juszczuk, Marek; Jerka-Dziadosz, Maria; Maliszewska, Kamila; Mucchielli, Marie-Hélène; Gout, Jean-François; Arnaiz, Olivier; Agier, Nicolas; Tang, Thomas; Aggerbeck, Lawrence P; Cohen, Jean; Delacroix, Hervé; Sperling, Linda; Herbert, Christopher J; Zagulski, Marek; Bétermier, Mireille

    2011-03-01

    Like all ciliates, Paramecium tetraurelia is a unicellular eukaryote that harbors two kinds of nuclei within its cytoplasm. At each sexual cycle, a new somatic macronucleus (MAC) develops from the germ line micronucleus (MIC) through a sequence of complex events, which includes meiosis, karyogamy, and assembly of the MAC genome from MIC sequences. The latter process involves developmentally programmed genome rearrangements controlled by noncoding RNAs and a specialized RNA interference machinery. We describe our first attempts to identify genes and biological processes that contribute to the progression of the sexual cycle. Given the high percentage of unknown genes annotated in the P. tetraurelia genome, we applied a global strategy to monitor gene expression profiles during autogamy, a self-fertilization process. We focused this pilot study on the genes carried by the largest somatic chromosome and designed dedicated DNA arrays covering 484 genes from this chromosome (1.2% of all genes annotated in the genome). Transcriptome analysis revealed four major patterns of gene expression, including two successive waves of gene induction. Functional analysis of 15 upregulated genes revealed four that are essential for vegetative growth, one of which is involved in the maintenance of MAC integrity and another in cell division or membrane trafficking. Two additional genes, encoding a MIC-specific protein and a putative RNA helicase localizing to the old and then to the new MAC, are specifically required during sexual processes. Our work provides a proof of principle that genes essential for meiosis and nuclear reorganization can be uncovered following genome-wide transcriptome analysis.

  2. Genome-wide evolutionary analysis of the noncoding RNA genes and noncoding DNA of Paramecium tetraurelia

    PubMed Central

    Chen, Chun-Long; Zhou, Hui; Liao, Jian-You; Qu, Liang-Hu; Amar, Laurence

    2009-01-01

    The compact genome of the unicellular eukaryote Paramecium tetraurelia contains noncoding DNA (ncDNA) distributed into >39,000 intergenic sequences and >90,000 introns of 390 base pairs (bp) and 25 bp on average, respectively. Here we analyzed the molecular features of the ncRNA genes, introns, and intergenic sequences of this genome. We mainly used computational programs and comparative genomics possible because the P. tetraurelia genome had formed throughout whole-genome duplications (WGDs). We characterized 417 5S rRNA, snRNA, snoRNA, SRP RNA, and tRNA putative genes, 415 of which map within intergenic sequences, and two, within introns. The evolution of these ncRNA genes appears to have mainly involved purifying selection and gene deletion. We then compared the introns that interrupt the protein-coding gene duplicates arisen from the recent WGD and identified a population of a few thousands of introns having evolved under most stringent constraints (>95% of identity). We also showed that low nucleotide substitution levels characterize the 50 and 80–115 base pairs flanking, respectively, the stop and start codons of the protein-coding genes. Lower substitution levels mark the base pairs flanking the highly transcribed genes, or the start codons of the genes of the sets with a high number of WGD-related sequences. Finally, adjacent to protein-coding genes, we characterized 32 DNA motifs able to encode stable and evolutionary conserved RNA secondary structures and defining putative expression controlling elements. Fourteen DNA motifs with similar properties map distant from protein-coding genes and may encode regulatory ncRNAs. PMID:19218550

  3. Paramecium caudatum enhances transmission and infectivity of Mycobacterium marinum and M. chelonae in zebrafish Danio rerio.

    PubMed

    Peterson, Tracy S; Ferguson, Jayde A; Watral, Virginia G; Mutoji, K Nadine; Ennis, Don G; Kent, Michael L

    2013-11-06

    Mycobacterial infections in laboratory zebrafish Danio rerio are common and widespread in research colonies. Mycobacteria within free-living amoebae have been shown to be transmission vectors for mycobacteriosis. Paramecium caudatum are commonly used as a first food for zebrafish, and we investigated this ciliate's potential to serve as a vector of Mycobacterium marinum and M. chelonae. The ability of live P. caudatum to transmit these mycobacteria to larval, juvenile and adult zebrafish was evaluated. Infections were defined by histologic observation of granulomas containing acid-fast bacteria in extraintestinal locations. In both experiments, fish fed paramecia containing mycobacteria became infected at a higher incidence than controls. Larvae (exposed at 4 d post hatch) fed paramecia with M. marinum exhibited an incidence of 30% (24/80) and juveniles (exposed at 21 d post hatch) showed 31% incidence (14/45). Adult fish fed a gelatin food matrix containing mycobacteria within paramecia or mycobacteria alone for 2 wk resulted in infections when examined 8 wk after exposure as follows: M. marinum OSU 214 47% (21/45), M. marinum CH 47% (9/19), and M. chelonae 38% (5/13). In contrast, fish feed mycobacteria alone in this diet did not become infected, except for 2 fish (5%) in the M. marinum OSU 214 low-dose group. These results demonstrate that P. caudatum can act as a vector for mycobacteria. This provides a useful animal model for evaluation of natural mycobacterial infections and demonstrates the possibility of mycobacterial transmission in zebrafish facilities via contaminated paramecia cultures.

  4. Secretory protein decondensation as a distinct, Ca2+-mediated event during the final steps of exocytosis in Paramecium cells

    PubMed Central

    1981-01-01

    The contents of secretory vesicles ("trichocysts") were isolated in the condensed state from Paramecium cells. It is well known that the majority portion of trichocysts perform a rapid decondensation process during exocytosis, which is visible in the light microscope. We have analyzed this condensed leads to decondensed transition in vitro and determined some relevant parameters. In the condensed state, free phosphate (and possibly magnesium) ions screen local surplus charges. This is supported by x-ray spectra recorded from individual trichocysts (prepared by physical methods) in a scanning transmission electron microscope. Calcium, as well as other ions that eliminate phosphate by precipitation, produces decondensation in vitro. Under in vivo conditions, Ca2+ enters the vesicle lumen from the outside medium, once an exocytic opening has been formed. Consequently, within the intact cell, membrane fusion and protein decondensation take place with optimal timing. Ca2+ might then trigger decondensation in the same way by precipitating phosphate ions (as it does in vitro) and, indeed, such precipitates (again yielding Ca and P signals in x-ray spectra) can be recognized in situ under trigger conditions. As decondensation is a unidirectional, rapid process in Paramecium cells, it would contribute to drive the discharge of the secretory contents to the outside. Further implications on the energetics of exocytosis are discussed. PMID:7204486

  5. Identification, Localization, and Functional Implications of the Microdomain-Forming Stomatin Family in the Ciliated Protozoan Paramecium tetraurelia

    PubMed Central

    Stuermer, Claudia A. O.; Plattner, Helmut

    2013-01-01

    The SPFH protein superfamily is assumed to occur universally in eukaryotes, but information from protozoa is scarce. In the Paramecium genome, we found only Stomatins, 20 paralogs grouped in 8 families, STO1 to STO8. According to cDNA analysis, all are expressed, and molecular modeling shows the typical SPFH domain structure for all subgroups. For further analysis we used family-specific sequences for fluorescence and immunogold labeling, gene silencing, and functional tests. With all family members tested, we found a patchy localization at/near the cell surface and on vesicles. The Sto1p and Sto4p families are also associated with the contractile vacuole complex. Sto4p also makes puncta on some food vacuoles and is abundant on vesicles recycling from the release site of spent food vacuoles to the site of nascent food vacuole formation. Silencing of the STO1 family reduces mechanosensitivity (ciliary reversal upon touching an obstacle), thus suggesting relevance for positioning of mechanosensitive channels in the plasmalemma. Silencing of STO4 members increases pulsation frequency of the contractile vacuole complex and reduces phagocytotic activity of Paramecium cells. In summary, Sto1p and Sto4p members seem to be involved in positioning specific superficial and intracellular microdomain-based membrane components whose functions may depend on mechanosensation (extracellular stimuli and internal osmotic pressure). PMID:23376944

  6. Intraspecific differentiation of Paramecium novaurelia strains (Ciliophora, Protozoa) inferred from phylogenetic analysis of ribosomal and mitochondrial DNA variation.

    PubMed

    Tarcz, Sebastian

    2013-01-01

    Paramecium novaurelia Beale and Schneller, 1954, was first found in Scotland and is known to occur mainly in Europe, where it is the most common species of the P. aurelia complex. In recent years, two non-European localities have been described: Turkey and the United States of America. This article presents the analysis of intraspecific variability among 25 strains of P. novaurelia with the application of ribosomal and mitochondrial loci (ITS1-5.8S-ITS2, 5' large subunit rDNA (5'LSU rDNA) and cytochrome c oxidase subunit 1 (COI) mtDNA). The mean distance observed for all of the studied P. novaurelia sequence pairs was p=0.008/0.016/0.092 (ITS1-5.8S-ITS2/5'LSU rDNA/COI). Phylogenetic trees (NJ/MP/BI) based on a comparison of all of the analysed sequences show that the studied strains of P. novaurelia form a distinct clade, separate from the P. caudatum outgroup, and are divided into two clusters (A and B) and two branches (C and D). The occurrence of substantial genetic differentiation within P. novaurelia, confirmed by the analysed DNA fragments, indicates a rapid evolution of particular species within the Paramecium genus. Copyright © 2012 Elsevier GmbH. All rights reserved.

  7. Polymorphism of Paramecium pentaurelia (Ciliophora, Oligohymenophorea) strains revealed by rDNA and mtDNA sequences.

    PubMed

    Przyboś, Ewa; Tarcz, Sebastian; Greczek-Stachura, Magdalena; Surmacz, Marta

    2011-05-01

    Paramecium pentaurelia is one of 15 known sibling species of the Paramecium aurelia complex. It is recognized as a species showing no intra-specific differentiation on the basis of molecular fingerprint analyses, whereas the majority of other species are polymorphic. This study aimed at assessing genetic polymorphism within P. pentaurelia including new strains recently found in Poland (originating from two water bodies, different years, seasons, and clones of one strain) as well as strains collected from distant habitats (USA, Europe, Asia), and strains representing other species of the complex. We compared two DNA fragments: partial sequences (349 bp) of the LSU rDNA and partial sequences (618 bp) of cytochrome B gene. A correlation between the geographical origin of the strains and the genetic characteristics of their genotypes was not observed. Different genotypes were found in Kraków in two types of water bodies (Opatkowice-natural pond; Jordan's Park-artificial pond). Haplotype diversity within a single water body was not recorded. Likewise, seasonal haplotype differences between the strains within the artificial water body, as well as differences between clones originating from one strain, were not detected. The clustering of some strains belonging to different species was observed in the phylogenies. Copyright © 2010 Elsevier GmbH. All rights reserved.

  8. Early stages of functional diversification in the Rab GTPase gene family revealed by genomic and localization studies in Paramecium species

    PubMed Central

    Bright, Lydia J.; Gout, Jean-Francois; Lynch, Michael

    2017-01-01

    New gene functions arise within existing gene families as a result of gene duplication and subsequent diversification. To gain insight into the steps that led to the functional diversification of paralogues, we tracked duplicate retention patterns, expression-level divergence, and subcellular markers of functional diversification in the Rab GTPase gene family in three Paramecium aurelia species. After whole-genome duplication, Rab GTPase duplicates are more highly retained than other genes in the genome but appear to be diverging more rapidly in expression levels, consistent with early steps in functional diversification. However, by localizing specific Rab proteins in Paramecium cells, we found that paralogues from the two most recent whole-genome duplications had virtually identical localization patterns, and that less closely related paralogues showed evidence of both conservation and diversification. The functionally conserved paralogues appear to target to compartments associated with both endocytic and phagocytic recycling functions, confirming evolutionary and functional links between the two pathways in a divergent eukaryotic lineage. Because the functionally diversifying paralogues are still closely related to and derived from a clade of functionally conserved Rab11 genes, we were able to pinpoint three specific amino acid residues that may be driving the change in the localization and thus the function in these proteins. PMID:28251922

  9. Synchronous induction of detachment and reattachment of symbiotic Chlorella spp. from the cell cortex of the host Paramecium bursaria.

    PubMed

    Kodama, Yuuki; Fujishima, Masahiro

    2013-09-01

    Paramecium bursaria harbor several hundred symbiotic Chlorella spp. Each alga is enclosed in a perialgal vacuole membrane, which can attach to the host cell cortex. How the perialgal vacuole attaches beneath the host cell cortex remains unknown. High-speed centrifugation (> 1000×g) for 1min induces rapid detachment of the algae from the host cell cortex and concentrates the algae to the posterior half of the host cell. Simultaneously, most of the host acidosomes and lysosomes accumulate in the anterior half of the host cell. Both the detached algae and the dislocated acidic vesicles recover their original positions by host cyclosis within 10min after centrifugation. These recoveries were inhibited if the host cytoplasmic streaming was arrested by nocodazole. Endosymbiotic algae during the early reinfection process also show the capability of desorption after centrifugation. These results demonstrate that adhesion of the perialgal vacuole beneath the host cell cortex is repeatedly inducible, and that host cytoplasmic streaming facilitates recovery of the algal attachment. This study is the first report to illuminate the mechanism of the induction to desorb for symbiotic algae and acidic vesicles, and will contribute to the understanding of the mechanism of algal and organelle arrangements in Paramecium. Copyright © 2013 Elsevier GmbH. All rights reserved.

  10. Inactivation of Ca2+-induced ciliary reversal by high-salt extraction in the cilia of Paramecium.

    PubMed

    Kutomi, Osamu; Seki, Makoto; Nakamura, Shogo; Kamachi, Hiroyuki; Noguchi, Munenori

    2013-10-01

    Intracellular Ca(2+) induces ciliary reversal and backward swimming in Paramecium. However, it is not known how the Ca(2+) signal controls the motor machinery to induce ciliary reversal. We found that demembranated cilia on the ciliated cortical sheets from Paramecium caudatum lost the ability to undergo ciliary reversal after brief extraction with a solution containing 0.5 M KCl. KNO(3), which is similar to KCl with respect to chaotropic effect; it had the same effect as that of KCl on ciliary response. Cyclic AMP antagonizes Ca(2+)-induced ciliary reversal. Limited trypsin digestion prevents endogenous A-kinase and cAMP-dependent phosphorylation of an outer arm dynein light chain and induces ciliary reversal. However, the trypsin digestion prior to the high-salt extraction did not affect the inhibition of Ca(2+)-induced ciliary reversal caused by the high-salt extraction. Furthermore, during the course of the high-salt extraction, some axonemal proteins were extracted from ciliary axonemes, suggesting that they may be responsible for Ca(2+)-induced ciliary reversal.

  11. Paramecium: a promising non-animal bioassay to study the effect of 808 nm infrared diode laser photobiomodulation.

    PubMed

    Amaroli, Andrea; Parker, Steven; Dorigo, Gianluca; Benedicenti, Alberico; Benedicenti, Stefano

    2015-01-01

    Photobiostimulation and photobiomodulation (PBM) are terms applied to the manipulation of cellular behavior using low intensity light sources, which works on the principle of inducing a biological response through energy transfer. The aim of this investigation was to identify a laboratory assay to test the effect of an infrared diode laser light (808 nm) on cell fission rate. Sixty cells of Paramecium primaurelia were divided in two groups of 30. The first group (test group) was irradiated, at a temperature of 24°C, for 50 sec by a 808 nm diode laser with a flat top handpiece [1 cm of spot diameter, 1 W in continuous wave (CW), 50 sec irradiation time, 64 J/cm(2) of fluence]. The second group (control group) received no laser irradiation. All cells were transferred onto a depression slide, fed, and incubated in a moist chamber at a temperature of 24°C. The cells were exposed and monitored for 10 consecutive fission rates. Changes in temperature and pH were also evaluated. The exposed cells had a fission rate rhythm faster than the control cells, showing a binary fission significantly (p<0.05) shorter than unexposed cells. No significant effects of laser irradiation on pH and temperature of Paramecium's lettuce infusion medium were observed. The 808 nm infrared diode laser light, at the irradiation parameters used in our work, results in a precocious fission rate in P. primaurelia cells, probably through an increase in metabolic activity, secondary to an energy transfer.

  12. Functional specialization of Piwi proteins in Paramecium tetraurelia from post-transcriptional gene silencing to genome remodelling.

    PubMed

    Bouhouche, Khaled; Gout, Jean-François; Kapusta, Aurélie; Bétermier, Mireille; Meyer, Eric

    2011-05-01

    Proteins of the Argonaute family are small RNA carriers that guide regulatory complexes to their targets. The family comprises two major subclades. Members of the Ago subclade, which are present in most eukaryotic phyla, bind different classes of small RNAs and regulate gene expression at both transcriptional and post-transcriptional levels. Piwi subclade members appear to have been lost in plants and fungi and were mostly studied in metazoa, where they bind piRNAs and have essential roles in sexual reproduction. Their presence in ciliates, unicellular organisms harbouring both germline micronuclei and somatic macronuclei, offers an interesting perspective on the evolution of their functions. Here, we report phylogenetic and functional analyses of the 15 Piwi genes from Paramecium tetraurelia. We show that four constitutively expressed proteins are involved in siRNA pathways that mediate gene silencing throughout the life cycle. Two other proteins, specifically expressed during meiosis, are required for accumulation of scnRNAs during sexual reproduction and for programmed genome rearrangements during development of the somatic macronucleus. Our results indicate that Paramecium Piwi proteins have evolved to perform both vegetative and sexual functions through mechanisms ranging from post-transcriptional mRNA cleavage to epigenetic regulation of genome rearrangements.

  13. Functional specialization of Piwi proteins in Paramecium tetraurelia from post-transcriptional gene silencing to genome remodelling

    PubMed Central

    Bouhouche, Khaled; Gout, Jean-François; Kapusta, Aurélie; Bétermier, Mireille; Meyer, Eric

    2011-01-01

    Proteins of the Argonaute family are small RNA carriers that guide regulatory complexes to their targets. The family comprises two major subclades. Members of the Ago subclade, which are present in most eukaryotic phyla, bind different classes of small RNAs and regulate gene expression at both transcriptional and post-transcriptional levels. Piwi subclade members appear to have been lost in plants and fungi and were mostly studied in metazoa, where they bind piRNAs and have essential roles in sexual reproduction. Their presence in ciliates, unicellular organisms harbouring both germline micronuclei and somatic macronuclei, offers an interesting perspective on the evolution of their functions. Here, we report phylogenetic and functional analyses of the 15 Piwi genes from Paramecium tetraurelia. We show that four constitutively expressed proteins are involved in siRNA pathways that mediate gene silencing throughout the life cycle. Two other proteins, specifically expressed during meiosis, are required for accumulation of scnRNAs during sexual reproduction and for programmed genome rearrangements during development of the somatic macronucleus. Our results indicate that Paramecium Piwi proteins have evolved to perform both vegetative and sexual functions through mechanisms ranging from post-transcriptional mRNA cleavage to epigenetic regulation of genome rearrangements. PMID:21216825

  14. A scanning electron-microscopic study of the local degeneration of cilia during sexual reproduction in Paramecium.

    PubMed

    Watanabe, T

    1978-08-01

    The location and extent of local degeneration of cilia during sexual reproduction of Paramecium was studied using scanning electron microscopy to examine cells undergoing conjugation and autogamy. At some time during the mating reaction, but prior to conjugant pair formation, ciliary degeneration begins at the antero-ventral tip of cells and proceeds posteriorly along the suture. In the anterior part of the cell, degeneration occurs on both sides of the suture, but in the posterior part it is restricted to the right side of the suture. In 5 species of Paramecium examined, degeneration occurred in nearly the same region. No degeneration of cilia is observed in natural autogamy of P. tetraurelia, whereas in chemically induced autogamy of P. caudatum degeneration occurs as in ordinary conjugation. Conjugant pairs never expose any deciliated cell surface except at the postero-ventral tip. The maximum extent of ciliary degeneration is best seen in the chemically induced autogamous cells: 7 kinetics (rows of unit teritories) at the anterior-left, 4 kinetics at the anterior-right, 10 or more kinetics at the posterior-right and the right wall of the vestibule of the mouth. Before complete disappearance of the cilia, many short cilia are observed. This suggests that ciliary degeneration is due to resorption. Degeneration extends more rapidly in cells with stronger mating reactivity. The relations between mating reactivity, ciliary degeneration and nuclear activation are discussed.

  15. Primary and secondary siRNA synthesis triggered by RNAs from food bacteria in the ciliate Paramecium tetraurelia

    PubMed Central

    Carradec, Quentin; Götz, Ulrike; Arnaiz, Olivier; Pouch, Juliette; Simon, Martin; Meyer, Eric; Marker, Simone

    2015-01-01

    In various organisms, an efficient RNAi response can be triggered by feeding cells with bacteria producing double-stranded RNA (dsRNA) against an endogenous gene. However, the detailed mechanisms and natural functions of this pathway are not well understood in most cases. Here, we studied siRNA biogenesis from exogenous RNA and its genetic overlap with endogenous RNAi in the ciliate Paramecium tetraurelia by high-throughput sequencing. Using wild-type and mutant strains deficient for dsRNA feeding we found that high levels of primary siRNAs of both strands are processed from the ingested dsRNA trigger by the Dicer Dcr1, the RNA-dependent RNA polymerases Rdr1 and Rdr2 and other factors. We further show that this induces the synthesis of secondary siRNAs spreading along the entire endogenous mRNA, demonstrating the occurrence of both 3′-to-5′ and 5′-to-3′ transitivity for the first time in the SAR clade of eukaryotes (Stramenopiles, Alveolates, Rhizaria). Secondary siRNAs depend on Rdr2 and show a strong antisense bias; they are produced at much lower levels than primary siRNAs and hardly contribute to RNAi efficiency. We further provide evidence that the Paramecium RNAi machinery also processes single-stranded RNAs from its bacterial food, broadening the possible natural functions of exogenously induced RNAi in this organism. PMID:25593325

  16. Two isoforms of eukaryotic phospholipase C in Paramecium affecting transport and release of GPI-anchored proteins in vivo.

    PubMed

    Klöppel, Christine; Müller, Alexandra; Marker, Simone; Simon, Martin

    2009-10-01

    Surface proteins anchored by a glycosylphosphatidylinositol (GPI) residue in the cell membrane are widely distributed among eukaryotic cells. The GPI anchor is cleavable by a phospholipase C (PLC) leading to the release of such surface proteins, and this process is postulated to be essential in several systems. For higher eukaryotes, the responsible enzymes have not been characterized in any detail as yet. Here we characterize six PLCs in the ciliated protozoan, Paramecium, which, in terms of catalytic domains and architecture, all show characteristics of PLCs involved in signal transduction in higher eukaryotes. We show that some of these endogenous PLCs can release GPI-anchored surface proteins in vitro: using RNA(i) to reduce PLC expression results in the same effects as the application of PLC inhibitors. With two enzymes, PLC2 and PLC6, RNA(i) phenotypes show strong defects in release of GPI-anchored surface proteins in vivo. Moreover, these RNA(i) lines also show abnormal surface protein distribution, suggesting that GPI cleavage may influence trafficking of anchored proteins. As we find GFP fusion proteins in the cytosol and in the surface protein extracts, these PLCs obviously show unconventional translocation mechanisms. This is the first molecular data on endogenous Paramecium PLCs with the described properties affecting GPI anchors in vitro and in vivo.

  17. Primary and secondary siRNA synthesis triggered by RNAs from food bacteria in the ciliate Paramecium tetraurelia.

    PubMed

    Carradec, Quentin; Götz, Ulrike; Arnaiz, Olivier; Pouch, Juliette; Simon, Martin; Meyer, Eric; Marker, Simone

    2015-02-18

    In various organisms, an efficient RNAi response can be triggered by feeding cells with bacteria producing double-stranded RNA (dsRNA) against an endogenous gene. However, the detailed mechanisms and natural functions of this pathway are not well understood in most cases. Here, we studied siRNA biogenesis from exogenous RNA and its genetic overlap with endogenous RNAi in the ciliate Paramecium tetraurelia by high-throughput sequencing. Using wild-type and mutant strains deficient for dsRNA feeding we found that high levels of primary siRNAs of both strands are processed from the ingested dsRNA trigger by the Dicer Dcr1, the RNA-dependent RNA polymerases Rdr1 and Rdr2 and other factors. We further show that this induces the synthesis of secondary siRNAs spreading along the entire endogenous mRNA, demonstrating the occurrence of both 3'-to-5' and 5'-to-3' transitivity for the first time in the SAR clade of eukaryotes (Stramenopiles, Alveolates, Rhizaria). Secondary siRNAs depend on Rdr2 and show a strong antisense bias; they are produced at much lower levels than primary siRNAs and hardly contribute to RNAi efficiency. We further provide evidence that the Paramecium RNAi machinery also processes single-stranded RNAs from its bacterial food, broadening the possible natural functions of exogenously induced RNAi in this organism.

  18. Tolerance of ciliated protozoan Paramecium bursaria (Protozoa, Ciliophora) to ammonia and nitrites

    NASA Astrophysics Data System (ADS)

    Xu, Henglong; Song, Weibo; Lu, Lu; Alan, Warren

    2005-09-01

    The tolerance to ammonia and nitrites in freshwater ciliate Paramecium bursaria was measured in a conventional open system. The ciliate was exposed to different concentrations of ammonia and nitrites for 2h and 12h in order to determine the lethal concentrations. Linear regression analysis revealed that the 2h-LC50 value for ammonia was 95.94 mg/L and for nitrite 27.35 mg/L using probit scale method (with 95% confidence intervals). There was a linear correlation between the mortality probit scale and logarithmic concentration of ammonia which fit by a regression equation y=7.32 x 9.51 ( R 2=0.98; y, mortality probit scale; x, logarithmic concentration of ammonia), by which 2 h-LC50 value for ammonia was found to be 95.50 mg/L. A linear correlation between mortality probit scales and logarithmic concentration of nitrite is also followed the regression equation y=2.86 x+0.89 ( R 2=0.95; y, mortality probit scale; x, logarithmic concentration of nitrite). The regression analysis of toxicity curves showed that the linear correlation between exposed time of ammonia-N LC50 value and ammonia-N LC50 value followed the regression equation y=2 862.85 e -0.08 x ( R 2=0.95; y, duration of exposure to LC50 value; x, LC50 value), and that between exposed time of nitrite-N LC50 value and nitrite-N LC50 value followed the regression equation y=127.15 e -0.13 x ( R 2=0.91; y, exposed time of LC50 value; x, LC50 value). The results demonstrate that the tolerance to ammonia in P. bursaria is considerably higher than that of the larvae or juveniles of some metozoa, e.g. cultured prawns and oysters. In addition, ciliates, as bacterial predators, are likely to play a positive role in maintaining and improving water quality in aquatic environments with high-level ammonium, such as sewage treatment systems.

  19. Highly Precise and Developmentally Programmed Genome Assembly in Paramecium Requires Ligase IV–Dependent End Joining

    PubMed Central

    Marmignon, Antoine; Ku, Michael; Silve, Aude; Meyer, Eric; Forney, James D.; Malinsky, Sophie; Bétermier, Mireille

    2011-01-01

    During the sexual cycle of the ciliate Paramecium, assembly of the somatic genome includes the precise excision of tens of thousands of short, non-coding germline sequences (Internal Eliminated Sequences or IESs), each one flanked by two TA dinucleotides. It has been reported previously that these genome rearrangements are initiated by the introduction of developmentally programmed DNA double-strand breaks (DSBs), which depend on the domesticated transposase PiggyMac. These DSBs all exhibit a characteristic geometry, with 4-base 5′ overhangs centered on the conserved TA, and may readily align and undergo ligation with minimal processing. However, the molecular steps and actors involved in the final and precise assembly of somatic genes have remained unknown. We demonstrate here that Ligase IV and Xrcc4p, core components of the non-homologous end-joining pathway (NHEJ), are required both for the repair of IES excision sites and for the circularization of excised IESs. The transcription of LIG4 and XRCC4 is induced early during the sexual cycle and a Lig4p-GFP fusion protein accumulates in the developing somatic nucleus by the time IES excision takes place. RNAi–mediated silencing of either gene results in the persistence of free broken DNA ends, apparently protected against extensive resection. At the nucleotide level, controlled removal of the 5′-terminal nucleotide occurs normally in LIG4-silenced cells, while nucleotide addition to the 3′ ends of the breaks is blocked, together with the final joining step, indicative of a coupling between NHEJ polymerase and ligase activities. Taken together, our data indicate that IES excision is a “cut-and-close” mechanism, which involves the introduction of initiating double-strand cleavages at both ends of each IES, followed by DSB repair via highly precise end joining. This work broadens our current view on how the cellular NHEJ pathway has cooperated with domesticated transposases for the emergence of new

  20. Three Group-I introns in 18S rDNA of Endosymbiotic Algae of Paramecium bursaria from Japan

    NASA Astrophysics Data System (ADS)

    Hoshina, Ryo; Kamako, Shin-ichiro; Imamura, Nobutaka

    2004-08-01

    In the nuclear encoded small subunit ribosomal DNA (18S rDNA) of symbiotic alga of Paramecium bursaria (F36 collected in Japan) possesses three intron-like insertions (Hoshina et al., unpubl. data, 2003). The present study confirmed these exact lengths and insertion sites by reverse transcription-PCR. Two of them were inserted at Escherichia coli 16S rRNA genic position 943 and 1512 that are frequent intron insertion positions, but another insertion position (nearly 1370) was the first finding. Their secondary structures suggested they belong to Group-I intron; one belongs to subgroup IE, others belong to subgroup IC1. Similarity search indicated these introns are ancestral ones.

  1. Phylogenetic placement of two previously described intranuclear bacteria from the ciliate Paramecium bursaria (Protozoa, Ciliophora): 'Holospora acuminata' and 'Holospora curviuscula'.

    PubMed

    Rautian, Maria S; Wackerow-Kouzova, Natalia D

    2013-05-01

    'Holospora acuminata' infects micronuclei of Paramecium bursaria (Protozoa, Ciliophora), whereas 'Holospora curviuscula' infects the macronucleus in other clones of the same host species. Because these micro-organisms have not been cultivated, their description has been based only on some morphological properties and host and nuclear specificities. One16S rRNA gene sequence of 'H. curviuscula' is present in databases. The systematic position of the representative strain of 'H. curviuscula', strain MC-3, was determined in this study. Moreover, for the first time, two strains of 'H. acuminata', KBN10-1 and AC61-10, were investigated. Phylogenetic analysis indicated that all three strains belonged to the genus Holospora, family Holosporaceae, order Rickettsiales within the Alphaproteobacteria.

  2. X-ray microanalysis in cryosections of natively frozen Paramecium caudatum with regard to ion distribution in ciliates

    SciTech Connect

    Schmitz, M.; Meyer, R.; Zierold, K.

    1985-01-01

    Cells of Paramecium caudatum were shock-frozen without pretreatment for cryoultramicrotomy and freeze-dried for subsequent X-ray microanalysis. Na, Mg, P, S, Cl, K, and Ca were detected in different amounts in several subcellular compartments. In particular, calcium was localized below the cell surface (pellicle). Trichocysts were found to contain significant amounts of Na in their base but not in the tip. Na, Mg, P, S, Cl, K, Ca were found in electron dense deposits within the lumen of the contractile vacuole. A small K concentration was found in the cytoplasm and in the mitochondria. X-ray microanalysis of the element distribution in different subcellular compartments provides information for the understanding of cellular functions such as exocytosis, locomotion, and ion regulation.

  3. Molecular Identification of Paramecium bursaria Syngens and Studies on Geographic Distribution using Mitochondrial Cytochrome C Oxidase Subunit I (COI).

    PubMed

    Zagata, Patrycja; Greczek-Stachura, Magdalena; Tarcz, Sebastian; Rautian, Maria

    2015-01-01

    Paramecium bursaria is composed of five syngens that are morphologically indistinguishable but sexually isolated. The aim of the present study was to confirm by molecular methods (analyses of mitochondrial COI) the identification of P. bursaria syngens originating from different geographical locations. Phylograms constructed using both the neighbor-joining and maximum-likelihood methods based on a comparison of 34 sequences of P. bursaria strains and P. multimicronucleatum, P. caudatum and P.calkinsi strains used as outgroups revealed five clusters which correspond to results obtained previously by mating reaction. Our analysis shows the existence of 24 haplotypes for the COI gene sequence in the studied strains. The interspecies haplotype diversity was Hd = 0.967. We confirmed genetic differentiation between strains of P. bursaria and the occurrence of a correlation between geographical distribution and the correspondent syngen.

  4. Commitment to autogamy in Paramecium blocks mating reactivity: implications for regulation of the sexual pathway and the breeding system.

    PubMed

    Berger, J D; Rahemtullah, S

    1990-03-01

    Commitment to autogamy blocks mating reactivity in Paramecium. Cells which had previously developed mating reactivity, lost reactivity 30-90 min prior to the preautogamous fission. Mating reactivity develops at a standard level of starvation when cells are allowed to exhaust their food supply naturally. In abruptly starved cultures, mating reactivity appears 3.3 h after downshift. Autogamy is also triggered by starvation. The level of starvation required for initiation of autogamy decreases progressively as cells age. When the autogamy starvation threshold drops to such a low level that all cells become committed to autogamy before any of them develop mating reactivity, reactivity does not occur under natural starvation conditions and the period of maturity for conjugation has come to an end. There is no absolute immature period for autogamy.

  5. Early stages of functional diversification in the Rab GTPase gene family revealed by genomic and localization studies in Paramecium species.

    PubMed

    Bright, Lydia J; Gout, Jean-Francois; Lynch, Michael

    2017-04-15

    New gene functions arise within existing gene families as a result of gene duplication and subsequent diversification. To gain insight into the steps that led to the functional diversification of paralogues, we tracked duplicate retention patterns, expression-level divergence, and subcellular markers of functional diversification in the Rab GTPase gene family in three Paramecium aurelia species. After whole-genome duplication, Rab GTPase duplicates are more highly retained than other genes in the genome but appear to be diverging more rapidly in expression levels, consistent with early steps in functional diversification. However, by localizing specific Rab proteins in Paramecium cells, we found that paralogues from the two most recent whole-genome duplications had virtually identical localization patterns, and that less closely related paralogues showed evidence of both conservation and diversification. The functionally conserved paralogues appear to target to compartments associated with both endocytic and phagocytic recycling functions, confirming evolutionary and functional links between the two pathways in a divergent eukaryotic lineage. Because the functionally diversifying paralogues are still closely related to and derived from a clade of functionally conserved Rab11 genes, we were able to pinpoint three specific amino acid residues that may be driving the change in the localization and thus the function in these proteins. © 2017 Bright et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  6. Flow cytometry sorting of nuclei enables the first global characterization of Paramecium germline DNA and transposable elements.

    PubMed

    Guérin, Frédéric; Arnaiz, Olivier; Boggetto, Nicole; Denby Wilkes, Cyril; Meyer, Eric; Sperling, Linda; Duharcourt, Sandra

    2017-04-26

    DNA elimination is developmentally programmed in a wide variety of eukaryotes, including unicellular ciliates, and leads to the generation of distinct germline and somatic genomes. The ciliate Paramecium tetraurelia harbors two types of nuclei with different functions and genome structures. The transcriptionally inactive micronucleus contains the complete germline genome, while the somatic macronucleus contains a reduced genome streamlined for gene expression. During development of the somatic macronucleus, the germline genome undergoes massive and reproducible DNA elimination events. Availability of both the somatic and germline genomes is essential to examine the genome changes that occur during programmed DNA elimination and ultimately decipher the mechanisms underlying the specific removal of germline-limited sequences. We developed a novel experimental approach that uses flow cell imaging and flow cytometry to sort subpopulations of nuclei to high purity. We sorted vegetative micronuclei and macronuclei during development of P. tetraurelia. We validated the method by flow cell imaging and by high throughput DNA sequencing. Our work establishes the proof of principle that developing somatic macronuclei can be sorted from a complex biological sample to high purity based on their size, shape and DNA content. This method enabled us to sequence, for the first time, the germline DNA from pure micronuclei and to identify novel transposable elements. Sequencing the germline DNA confirms that the Pgm domesticated transposase is required for the excision of all ~45,000 Internal Eliminated Sequences. Comparison of the germline DNA and unrearranged DNA obtained from PGM-silenced cells reveals that the latter does not provide a faithful representation of the germline genome. We developed a flow cytometry-based method to purify P. tetraurelia nuclei to high purity and provided quality control with flow cell imaging and high throughput DNA sequencing. We identified 61

  7. Ku-mediated coupling of DNA cleavage and repair during programmed genome rearrangements in the ciliate Paramecium tetraurelia.

    PubMed

    Marmignon, Antoine; Bischerour, Julien; Silve, Aude; Fojcik, Clémentine; Dubois, Emeline; Arnaiz, Olivier; Kapusta, Aurélie; Malinsky, Sophie; Bétermier, Mireille

    2014-08-01

    During somatic differentiation, physiological DNA double-strand breaks (DSB) can drive programmed genome rearrangements (PGR), during which DSB repair pathways are mobilized to safeguard genome integrity. Because of their unique nuclear dimorphism, ciliates are powerful unicellular eukaryotic models to study the mechanisms involved in PGR. At each sexual cycle, the germline nucleus is transmitted to the progeny, but the somatic nucleus, essential for gene expression, is destroyed and a new somatic nucleus differentiates from a copy of the germline nucleus. In Paramecium tetraurelia, the development of the somatic nucleus involves massive PGR, including the precise elimination of at least 45,000 germline sequences (Internal Eliminated Sequences, IES). IES excision proceeds through a cut-and-close mechanism: a domesticated transposase, PiggyMac, is essential for DNA cleavage, and DSB repair at excision sites involves the Ligase IV, a specific component of the non-homologous end-joining (NHEJ) pathway. At the genome-wide level, a huge number of programmed DSBs must be repaired during this process to allow the assembly of functional somatic chromosomes. To understand how DNA cleavage and DSB repair are coordinated during PGR, we have focused on Ku, the earliest actor of NHEJ-mediated repair. Two Ku70 and three Ku80 paralogs are encoded in the genome of P. tetraurelia: Ku70a and Ku80c are produced during sexual processes and localize specifically in the developing new somatic nucleus. Using RNA interference, we show that the development-specific Ku70/Ku80c heterodimer is essential for the recovery of a functional somatic nucleus. Strikingly, at the molecular level, PiggyMac-dependent DNA cleavage is abolished at IES boundaries in cells depleted for Ku80c, resulting in IES retention in the somatic genome. PiggyMac and Ku70a/Ku80c co-purify as a complex when overproduced in a heterologous system. We conclude that Ku has been integrated in the Paramecium DNA cleavage

  8. Ku-Mediated Coupling of DNA Cleavage and Repair during Programmed Genome Rearrangements in the Ciliate Paramecium tetraurelia

    PubMed Central

    Marmignon, Antoine; Bischerour, Julien; Silve, Aude; Fojcik, Clémentine; Dubois, Emeline; Arnaiz, Olivier; Kapusta, Aurélie; Malinsky, Sophie; Bétermier, Mireille

    2014-01-01

    During somatic differentiation, physiological DNA double-strand breaks (DSB) can drive programmed genome rearrangements (PGR), during which DSB repair pathways are mobilized to safeguard genome integrity. Because of their unique nuclear dimorphism, ciliates are powerful unicellular eukaryotic models to study the mechanisms involved in PGR. At each sexual cycle, the germline nucleus is transmitted to the progeny, but the somatic nucleus, essential for gene expression, is destroyed and a new somatic nucleus differentiates from a copy of the germline nucleus. In Paramecium tetraurelia, the development of the somatic nucleus involves massive PGR, including the precise elimination of at least 45,000 germline sequences (Internal Eliminated Sequences, IES). IES excision proceeds through a cut-and-close mechanism: a domesticated transposase, PiggyMac, is essential for DNA cleavage, and DSB repair at excision sites involves the Ligase IV, a specific component of the non-homologous end-joining (NHEJ) pathway. At the genome-wide level, a huge number of programmed DSBs must be repaired during this process to allow the assembly of functional somatic chromosomes. To understand how DNA cleavage and DSB repair are coordinated during PGR, we have focused on Ku, the earliest actor of NHEJ-mediated repair. Two Ku70 and three Ku80 paralogs are encoded in the genome of P. tetraurelia: Ku70a and Ku80c are produced during sexual processes and localize specifically in the developing new somatic nucleus. Using RNA interference, we show that the development-specific Ku70/Ku80c heterodimer is essential for the recovery of a functional somatic nucleus. Strikingly, at the molecular level, PiggyMac-dependent DNA cleavage is abolished at IES boundaries in cells depleted for Ku80c, resulting in IES retention in the somatic genome. PiggyMac and Ku70a/Ku80c co-purify as a complex when overproduced in a heterologous system. We conclude that Ku has been integrated in the Paramecium DNA cleavage

  9. Trichocysts-Paramecium's Projectile-like Secretory Organelles: Reappraisal of their Biogenesis, Composition, Intracellular Transport, and Possible Functions.

    PubMed

    Plattner, Helmut

    2017-01-01

    This review summarizes biogenesis, composition, intracellular transport, and possible functions of trichocysts. Trichocyst release by Paramecium is the fastest dense core-secretory vesicle exocytosis known. This is enabled by the crystalline nature of the trichocyst "body" whose matrix proteins (tmp), upon contact with extracellular Ca(2+) , undergo explosive recrystallization that propagates cooperatively throughout the organelle. Membrane fusion during stimulated trichocyst exocytosis involves Ca(2+) mobilization from alveolar sacs and tightly coupled store-operated Ca(2+) -influx, initiated by activation of ryanodine receptor-like Ca(2+) -release channels. Particularly, aminoethyldextran perfectly mimics a physiological function of trichocysts, i.e. defense against predators, by vigorous, local trichocyst discharge. The tmp's contained in the main "body" of a trichocyst are arranged in a defined pattern, resulting in crossstriation, whose period expands upon expulsion. The second part of a trichocyst, the "tip", contains secretory lectins which diffuse upon discharge. Repulsion from predators may not be the only function of trichocysts. We consider ciliary reversal accompanying stimulated trichocyst exocytosis (also in mutants devoid of depolarization-activated Ca(2+) channels) a second, automatically superimposed defense mechanism. A third defensive mechanism may be effectuated by the secretory lectins of the trichocyst tip; they may inhibit toxicyst exocytosis in Dileptus by crosslinking surface proteins (an effect mimicked in Paramecium by antibodies against cell surface components). Some of the proteins, body and tip, are glycosylated as visualized by binding of exogenous lectins. This reflects the biogenetic pathway, from the endoplasmic reticulum via the Golgi apparatus, which is also supported by details from molecular biology. There are fragile links connecting the matrix of a trichocyst with its membrane; these may signal the filling state, full or

  10. Structural studies demonstrating a bacteriophage-like replication cycle of the eukaryote-infecting Paramecium bursaria chlorella virus-1

    PubMed Central

    Shimoni, Eyal; Dadosh, Tali; Rechav, Katya; Unger, Tamar

    2017-01-01

    A fundamental stage in viral infection is the internalization of viral genomes in host cells. Although extensively studied, the mechanisms and factors responsible for the genome internalization process remain poorly understood. Here we report our observations, derived from diverse imaging methods on genome internalization of the large dsDNA Paramecium bursaria chlorella virus-1 (PBCV-1). Our studies reveal that early infection stages of this eukaryotic-infecting virus occurs by a bacteriophage-like pathway, whereby PBCV-1 generates a hole in the host cell wall and ejects its dsDNA genome in a linear, base-pair-by-base-pair process, through a membrane tunnel generated by the fusion of the virus internal membrane with the host membrane. Furthermore, our results imply that PBCV-1 DNA condensation that occurs shortly after infection probably plays a role in genome internalization, as hypothesized for the infection of some bacteriophages. The subsequent perforation of the host photosynthetic membranes presumably enables trafficking of viral genomes towards host nuclei. Previous studies established that at late infection stages PBCV-1 generates cytoplasmic organelles, termed viral factories, where viral assembly takes place, a feature characteristic of many large dsDNA viruses that infect eukaryotic organisms. PBCV-1 thus appears to combine a bacteriophage-like mechanism during early infection stages with a eukaryotic-like infection pathway in its late replication cycle. PMID:28850602

  11. Paramecium tetraurelia growth stimulation under low-level chronic irradiation: investigations on a possible mechanism. [/sup 60/Co

    SciTech Connect

    Croute, F.; Soleilhavoup, J.P.; Vidal, S.; Dupouy, D.; Planel, H.

    1982-12-01

    Experiments were carried out to demonstrate the effects of low-level chronic irradiation on Paramecium tetraurelia proliferation. Biological effects were strongly dependent on the bacterial density of culture medium and more exactly on the catalase content of the medium. Significant growth stimulation was found under /sup 60/Co chronic irradiation at a dose rate of 2 rad/year when paramecia were grown in a medium containing a high bacterial concentration (2.5 x 10/sup 2/ cells/m) or supplemented with catalase (300 U/ml). In a medium with a low bacterial density (1 x 10/sup 6/ cell/ml) or supplemented with a catalase activity inhibitor, growth simulation was preceded by a transitory inhibiting effect which could be correlated with extracellularly radioproduced H/sub 2/O/sub 2/. H/sub 2/O/sub 2/ addition appeared to be able to simulate the biological effects of chronic irradiation. A possible mechanism is discussed.We proposed that the stimulating effects were the result of intracellular enzymatic scavenging of radioproduced H/sub 2/O/sub 2/.

  12. Stage-specific appearance of cytoplasmic microtubules around the surviving nuclei during the third prezygotic division of Paramecium.

    PubMed

    Wang, Yi-Wen; Yuan, Jin-Qiang; Gao, Xin; Yang, Xian-Yu

    2012-12-01

    There are six micronuclear divisions during conjugation of Paramecium caudatum: three prezygotic and three postzygotic divisions. Four haploid nuclei are formed during the first two meiotic prezygotic divisions. Usually only one meiotic product is located in the paroral cone (PC) region at the completion of meiosis, which survives and divides mitotically to complete the third prezygotic division to yield a stationary and a migratory pronucleus. The remaining three located outside of the PC degenerate. The migratory pronuclei are then exchanged between two conjugants and fuse with the stationary pronuclei to form synkarya, which undergo three successive divisions (postzygotic divisions). However, little is known about the surviving mechanism of the PC nuclei. In the current study, stage-specific appearance of cytoplasmic microtubules (cMTs) was indicated during the third prezygotic division by immunofluorescence labeling with anti-alpha tubulin antibodies surrounding the surviving nuclei, including the PC nuclei and the two types of prospective pronuclei. This suggested that cMTs were involved in the formation of a physical barrier, whose function may relate to sequestering and protecting the surviving nuclei from the major cytoplasm, where degeneration of extra-meiotic products occurs, another important nuclear event during the third prezygotic division.

  13. Variation in ribosomal and mitochondrial DNA sequences demonstrates the existence of intraspecific groups in Paramecium multimicronucleatum (Ciliophora, Oligohymenophorea).

    PubMed

    Tarcz, Sebastian; Potekhin, Alexey; Rautian, Maria; Przyboś, Ewa

    2012-05-01

    This is the first phylogenetic study of the intraspecific variability within Paramecium multimicronucleatum with the application of two-loci analysis (ITS1-5.8S-ITS2-5'LSU rDNA and COI mtDNA) carried out on numerous strains originated from different continents. The species has been shown to have a complex structure of several sibling species within taxonomic species. Our analysis revealed the existence of 10 haplotypes for the rDNA fragment and 15 haplotypes for the COI fragment in the studied material. The mean distance for all of the studied P. multimicronucleatum sequence pairs was p=0.025/0.082 (rDNA/COI). Despite the greater variation of the COI fragment, the COI-derived tree topology is similar to the tree topology constructed on the basis of the rDNA fragment. P. multimicronucleatum strains are divided into three main clades. The tree based on COI fragment analysis presents a greater resolution of the studied P. multimicronucleatum strains. Our results indicate that the strains of P. multimicronucleatum that appear in different clades on the trees could belong to different syngens. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Basal body duplication in Paramecium: the key role of Bld10 in assembly and stability of the cartwheel.

    PubMed

    Jerka-Dziadosz, Maria; Gogendeau, Delphine; Klotz, Catherine; Cohen, Jean; Beisson, Janine; Koll, France

    2010-03-01

    Basal bodies which nucleate cilia and flagella, and centrioles which organize centrosomes share the same architecture characterized by the ninefold symmetry of their microtubular shaft. Among the conserved proteins involved in the biogenesis of the canonical 9-triplet centriolar structures, Sas-6 and Bld10 proteins have been shown to play central roles in the early steps of assembly and in establishment/stabilization of the ninefold symmetry. Using fluorescent tagged proteins and RNAi to study the localization and function of these two proteins in Paramecium, we focused on the early effects of their depletion, the consequences of their overexpression and their functional interdependence. We find that both genes are essential and their depletion affects cartwheel assembly and hence basal body duplication. We also show that, contrary to Sas6p, Bld10p is not directly responsible for the establishment of the ninefold symmetry, but is required not only for new basal body assembly and stability but also for Sas6p maintenance at mature basal bodies. Finally, ultrastructural analysis of cells overexpressing either protein revealed two types of early assembly intermediates, hub-like structures and generative discs, suggesting a conserved scaffolding process. (c) 2010 Wiley-Liss, Inc.

  15. Symbiotic Chlorella vulgaris of the ciliate Paramecium bursaria plays an important role in maintaining perialgal vacuole membrane functions.

    PubMed

    Kodama, Yuuki; Inouye, Isao; Fujishima, Masahiro

    2011-04-01

    Treatment of symbiotic alga-bearing Paramecium bursaria cells with a protein synthesis inhibitor, cycloheximide, induces synchronous swelling of all perialgal vacuoles at about 24h after treatment under a constant light condition. Subsequently, the vacuoles detach from the host cell cortex. The algae in the vacuoles are digested by the host's lysosomal fusion to the vacuoles. To elucidate the timing of algal degeneration, P. bursaria cells were treated with cycloheximide under a constant light condition. Then the cells were observed using transmission electron microscopy. Results show that algal chloroplasts and nuclei degenerated within 9h after treatment, but before the synchronous swelling of the perialgal vacuole and appearance of acid phosphatase activity in the perialgal vacuole by lysosomal fusion. Treatment with cycloheximide under a constant dark condition and treatment with chloramphenicol under a constant light condition induced neither synchronous swelling of the vacuoles nor digestion of the algae inside the vacuoles. These results demonstrate that algal proteins synthesized during photosynthesis are necessary to maintain chloroplastic and nuclear structures, and that inhibition of protein synthesis induces rapid lysis of these organelles, after which synchronous swelling of the perialgal vacuole and fusion occur with the host lysosomes. Copyright © 2010 Elsevier GmbH. All rights reserved.

  16. The first European stand of Paramecium sonneborni (P. aurelia complex), a species known only from North America (Texas, USA).

    PubMed

    Przyboś, Ewa; Tarcz, Sebastian; Rautian, Maria; Lebedeva, Natalia

    2014-06-01

    P. aurelia is currently defined as a complex of 15 sibling species including 14 species designated by Sonneborn (1975) and one, P. sonneborni, by Aufderheide et al. (1983). The latter was known from only one stand (Texas, USA). The main reason for the present study was a new stand of Paramecium in Cyprus, with strains recognized as P. sonneborni based on the results of strain crosses, cytological slides, and molecular analyses of three loci (ITS1-5.8S-ITS2-5'LSU rDNA, COI, CytB). The new stand of P. sonneborni in Europe shows that the species, previously considered endemic, may have a wider range. This demonstrates the impact of under-sampling on the knowledge of the biogeography of microbial eukaryotes. Phylogenetic trees based on all the studied fragments revealed that P. sonneborni forms a separate cluster that is closer to P. jenningsi and P. schewiakoffi than to the other members of the P. aurelia complex. Copyright © 2014 Elsevier GmbH. All rights reserved.

  17. Developmentally regulated excision of a 28-base-pair sequence from the Paramecium genome requires flanking DNA.

    PubMed

    Ku, M; Mayer, K; Forney, J D

    2000-11-01

    The micronuclear DNA of Paramecium tetraurelia is estimated to contain over 50,000 short DNA elements that are precisely removed during the formation of the transcriptionally active macronucleus. Each internal eliminated sequence (IES) is bounded by 5'-TA-3' dinucleotide repeats, a feature common to some classes of DNA transposons. We have developed an in vivo assay to analyze these highly efficient and precise DNA excision events. The microinjection of a cloned IES into mating cells results in accurately spliced products, and the transformed cells maintain the injected DNA as extrachromosomal molecules. A series of deletions flanking one side of a 28-bp IES were constructed and analyzed with the in vivo assay. Whereas 72 bp of DNA flanking the eliminated region is sufficient for excision, lengths of 31 and 18 bp result in reduced excision and removal of all wild-type sequences adjacent to the TA results in complete failure of excision. In contrast, nucleotide mutations within the middle of the 28-bp IES do not prevent excision. The results are consistent with a functional role for perfect inverted repeats flanking the IES.

  18. Evidence for the presence of a mammalian-like cholinesterase in Paramecium primaurelia (Protista, Ciliophora) developmental cycle.

    PubMed

    Delmonte Corrado, M U; Politi, H; Trielli, F; Angelini, C; Falugi, C

    1999-01-01

    By histochemical and immunohistochemical methods, the presence of cholinergic-like molecules has previously been demonstrated in Paramecium primaurelia, and their functional role in mating-cell pairing was suggested. In this work, both true acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities were electrophoretically investigated, and the presence of molecules immunologically related to BuChE was checked by immunoblotting. The AChE activity, shown in the membrane protein fraction of mating-competent cells and in the cytoplasmic fraction of immature cells, is due to a 260-kDa molecular form, similar to the membrane-bound tetrameric form present in human erythrocytes. This AChE activity does not appear in either the cytoplasmic fraction of mating-competent cells or in the membrane protein fraction of immature cells. No evidence was found for the presence or the activity of BuChE-like molecules. The role of AChE in P. primaurelia developmental cycle is discussed.

  19. Odd mating-type substances may work as precursor molecules of even mating-type substances in Paramecium caudatum.

    PubMed

    Xu, X; Kumakura, M; Kaku, E; Takahashi, M

    2001-01-01

    Mating-type substances are key molecules in the sexual recognition of the odd (O) and even (E) complementary mating-type cells in Paramecium caudatum. Indirect evidence suggested that the substances were proteins and were located on ventral surface cilia. Monoclonal antibodies inhibiting the mating reactivity of the O cells have been obtained. Using these antibodies, we tried to detect antigen molecules as dot-blot signals. Strong dot-blot signals of antigens were only detected from the mating reactive cells, but they were not detected from the well-fed and starved cells without mating reactivity. In addition to identifying the antigen on cilia and cytoplasm of the O cell, the antigen was detected from the cytoplasm of the E cells but never from their cilia. Furthermore, extracts of the E cells induced mating reaction with the living E cells but not with O cells. Thus, the O mating-type substances exist in the cytoplasm of the E mating-type cells, supporting strongly the hypothesis that O mating-type substances are precursor molecules of the E mating-type substances.

  20. Developmentally Regulated Excision of a 28-Base-Pair Sequence from the Paramecium Genome Requires Flanking DNA†

    PubMed Central

    Ku, Michael; Mayer, Kimberly; Forney, James D.

    2000-01-01

    The micronuclear DNA of Paramecium tetraurelia is estimated to contain over 50,000 short DNA elements that are precisely removed during the formation of the transcriptionally active macronucleus. Each internal eliminated sequence (IES) is bounded by 5′-TA-3′ dinucleotide repeats, a feature common to some classes of DNA transposons. We have developed an in vivo assay to analyze these highly efficient and precise DNA excision events. The microinjection of a cloned IES into mating cells results in accurately spliced products, and the transformed cells maintain the injected DNA as extrachromosomal molecules. A series of deletions flanking one side of a 28-bp IES were constructed and analyzed with the in vivo assay. Whereas 72 bp of DNA flanking the eliminated region is sufficient for excision, lengths of 31 and 18 bp result in reduced excision and removal of all wild-type sequences adjacent to the TA results in complete failure of excision. In contrast, nucleotide mutations within the middle of the 28-bp IES do not prevent excision. The results are consistent with a functional role for perfect inverted repeats flanking the IES. PMID:11046136

  1. Identification and characterization of a 38 kDa glycoprotein functionally associated with mating activity of Paramecium primaurelia.

    PubMed

    Ognibene, Marzia; Della Giovampaola, Cinzia; Trielli, Francesca; Focarelli, Riccardo; Rosati, Floriana; Umberta Delmonte Corrado, Maria

    2008-05-01

    In Paramecium primaurelia mating interactions take place immediately after mixing mating-competent cells of opposite mating types. The cells clump in clusters (mating reaction) and then separate in pairs. Previous results have shown that sialic acid-containing glycoconjugates are present on the cell surface and are involved in mating-cell pairing. In order to identify the sialic acid-containing glycoprotein(s), we first metabolically radiolabelled non-mating-competent cells with D-[6-(3)H]galactose, and then analyzed the radiolabelled proteins by anion exchange chromatography. We characterized a 38 kDa (gp38) sialic acid-containing glycoprotein and raised the corresponding polyclonal antibody by means of which we localized the antigen at the level of the oral region of non-mating-competent cells and on the ciliary surface of mating-competent cells. Immunoblot analysis of the ciliary protein fraction showed that the anti-gp38 serum interacted with a 38 kDa protein in both mating types I and II cells. We also demonstrated the functional activity of gp38 in the mating reaction by means of anti-gp38 antibody competition assays.

  2. Outer Dynein Arm Light Chain 1 Is Essential for Controlling the Ciliary Response to Cyclic AMP in Paramecium tetraurelia

    PubMed Central

    Kutomi, Osamu; Hori, Manabu; Ishida, Masaki; Tominaga, Takashi; Kamachi, Hiroyuki; Koll, France; Cohen, Jean; Yamada, Norico

    2012-01-01

    The individual role of the outer dynein arm light chains in the molecular mechanisms of ciliary movements in response to second messengers, such as Ca2+ and cyclic nucleotides, is unclear. We examined the role of the gene termed the outer dynein arm light chain 1 (LC1) gene of Paramecium tetraurelia (ODAL1), a homologue of the outer dynein arm LC1 gene of Chlamydomonas reinhardtii, in ciliary movements by RNA interference (RNAi) using a feeding method. The ODAL1-silenced (ODAL1-RNAi) cells swam slowly, and their swimming velocity did not increase in response to membrane-hyperpolarizing stimuli. Ciliary movements on the cortical sheets of ODAL1-RNAi cells revealed that the ciliary beat frequency was significantly lower than that of control cells in the presence of ≥1 mM Mg2+-ATP. In addition, the ciliary orientation of ODAL1-RNAi cells did not change in response to cyclic AMP (cAMP). A 29-kDa protein phosphorylated in a cAMP-dependent manner in the control cells disappeared in the axoneme of ODAL1-RNAi cells. These results indicate that ODAL1 is essential for controlling the ciliary response by cAMP-dependent phosphorylation. PMID:22427431

  3. Temperature-dependent transmission and latency of Holospora undulata, a micronucleus-specific parasite of the ciliate Paramecium caudatum

    PubMed Central

    Fels, Daniel; Kaltz, Oliver

    2006-01-01

    Transmission of parasites to new hosts crucially depends on the timing of production of transmission stages and their capacity to start an infection. These parameters may be influenced by genetic factors, but also by the environment. We tested the effects of temperature and host genotype on infection probability and latency in experimental populations of the ciliate Paramecium caudatum, after exposure to infectious forms of its bacterial parasite Holospora undulata. Temperature had a significant effect on the expression of genetic variation for transmission and maintenance of infection. Overall, low temperature (10 °C) increased levels of (multiple) infection, but arrested parasite development; higher temperatures (23 and 30 °C) accelerated the onset of production of infectious forms, but limited transmission success. Viability of infectious forms declined rapidly at 23 and 30 °C, thereby narrowing the time window for transmission. Thus, environmental conditions can generate trade-offs between transmission relevant parameters and alter levels of multiple infection or parasite-mediated selection, which may affect evolutionary trajectories of parasite life history or virulence. PMID:16627290

  4. Purification, in vitro reassembly, and preliminary sequence analysis of epiplasmins, the major constituent of the membrane skeleton of Paramecium.

    PubMed

    Coffe, G; Le Caer, J P; Lima, O; Adoutte, A

    1996-01-01

    The epiplasmic layer, a continuous rigid granulo-fibrillar sheet directly subtending the surface membranes of Paramecium, is one of the outermost of the various cytoskeletal networks that compose it cortex. We have previously shown that the epiplasm consists of a set of 30 to 50 protein bands on SDS-PAGE in the range 50 to 33 kDa, the epiplasmins. We report a purification procedure for the set of epiplasmic proteins, a description of their physicochemical and reassembly properties, and a preliminary characterization of their sequence. The conditions for solubilization of the epiplasm and for in vitro reassembly of its purified constituents ar described. Reassembly of the entire set of proteins and of some (but not all) subsets are shown to yield filamentous aggregates. Microsequences of two purified bands of epiplasmins reveal a striking amino acid sequence consisting of heptad repeats of only three main amino acids, P, V, and Q. These repeats were confirmed by DNA sequencing of polymerase chain reaction products. The motif is QPVQ-h, in which h is a hydrophobic residue. This may constitute the core of the epiplasmin sequence and, in view of the tendency of such a sequence to form a coiled-coil, may account for the remarkable self-aggregation properties of epiplasmins.

  5. The mechanism of the nephridial apparatus of Paramecium multimicronucleatum. II. The filling of the vesicle by action of the ampullae.

    PubMed

    Organ, A E; Bovee, E C; Jahn, T L

    1969-02-01

    Our recent analysis of the nephridial apparatus of Paramecium multimicronucleatum by high-speed cinematography (300 fps at X 250) indicates that before the water expulsion vesicle ("contractile vacuole") is completely voided of fluid during expulsion, the ampullae surrounding and confluent with the vesicle swell with fluid entering from their respective nephridial tubules. Once the membranes of the excretory pore at the base of the excretory canal (leading from the vesicle proper to the outside) have constricted and resealed the excretory pore, the up till then constricted injection tubules of the ampullae which conduct fluid to the vesicle open as waves of contraction along the coacervate gel around the ampulla and proceed along each ampulla from distal to proximal end. The coacervate gel around any one ampulla does not necessarily contract in phase with that of any other ampulla. Each ampulla acts independently. The fluid from the ampullae is thus pumped sequentially, but not in predetermined order, into the water expulsion vesicle, refilling and distending it. Our previous studies (Organ et al., 1968a) suggest that an actomyosinoid ATP-using mechanism may be functional in the ampullary contractions.

  6. Phylogenetically close group I introns with different positions among Paramecium bursaria photobionts imply a primitive stage of intron diversification.

    PubMed

    Hoshina, Ryo; Imamura, Nobutaka

    2009-06-01

    Group I introns are a distinct RNA group that catalyze their excision from precursor RNA transcripts and ligate the exons. Group I introns have a sporadic and highly biased distribution due to the two intron transfer mechanisms of homing and reverse splicing. These transfer pathways recognize assigned sequences even when introns are transferred beyond the species level. Consequently, introns at homologous gene sites between different host organisms are more related than those at heterologous sites within an organism. We describe the subgroup IE introns of two Chlorella species that are symbiotic green algae (photobionts) of a ciliate, Paramecium bursaria. One strain Chlorella sp. SW1-ZK (Csw.) had two IE introns at S651 and L2449, and the other strain Chlorella sp. OK1-ZK (Cok.) had four IE introns at S943, L1688, L1926, and L2184 (numbering reflects their homologous position in Escherichia coli rRNA gene: S = small subunit rRNA, L = large subunit rRNA). Despite locating on six heterologous sites, the introns formed a monophyletic clade independent of other groups. Phylogenetic and structural analyses of the introns indicated that Csw.L2449 has an archaic state, and the other introns are assumed to be originated from this intron. Some of the introns shared common internal guide sequences, which are necessary for misdirected transfer (i.e., transposition) via reverse splicing. Other introns, however, shared similar sequence fragments further upstream, after the insertions. We propose a hypothetical model to explain how these intron transpositions may have occurred in these photobionts; they transposed by a combination of homing-like event requiring relaxed sequence homology of recognition sequences and reverse splicing. This case study may represent a key to describe how group I intron explores new insertion sites.

  7. Coping with Temperature at the Warm Edge – Patterns of Thermal Adaptation in the Microbial Eukaryote Paramecium caudatum

    PubMed Central

    Krenek, Sascha; Petzoldt, Thomas; Berendonk, Thomas U.

    2012-01-01

    Background Ectothermic organisms are thought to be severely affected by global warming since their physiological performance is directly dependent on temperature. Latitudinal and temporal variations in mean temperatures force ectotherms to adapt to these complex environmental conditions. Studies investigating current patterns of thermal adaptation among populations of different latitudes allow a prediction of the potential impact of prospective increases in environmental temperatures on their fitness. Methodology/Principal Findings In this study, temperature reaction norms were ascertained among 18 genetically defined, natural clones of the microbial eukaryote Paramecium caudatum. These different clones have been isolated from 12 freshwater habitats along a latitudinal transect in Europe and from 3 tropical habitats (Indonesia). The sensitivity to increasing temperatures was estimated through the analysis of clone specific thermal tolerances and by relating those to current and predicted temperature data of their natural habitats. All investigated European clones seem to be thermal generalists with a broad thermal tolerance and similar optimum temperatures. The weak or missing co-variation of thermal tolerance with latitude does not imply local adaptation to thermal gradients; it rather suggests adaptive phenotypic plasticity among the whole European subpopulation. The tested Indonesian clones appear to be locally adapted to the less variable, tropical temperature regime and show higher tolerance limits, but lower tolerance breadths. Conclusions/Significance Due to the lack of local temperature adaptation within the European subpopulation, P. caudatum genotypes at the most southern edge of their geographic range seem to suffer from the predicted increase in magnitude and frequency of summer heat waves caused by climate change. PMID:22427799

  8. Identification of Two Nickel Ion-Induced Genes, NCI16 and PcGST1, in Paramecium caudatum

    PubMed Central

    Haga, Nobuyuki; Nakano, Takanari; Ikeda, Masaaki; Katayama, Shigehiro; Awata, Takuya

    2014-01-01

    Here, we describe the isolation of two nickel-induced genes in Paramecium caudatum, NCI16 and PcGST1, by subtractive hybridization. NCI16 encoded a predicted four-transmembrane domain protein (∼16 kDa) of unknown function, and PcGST1 encoded glutathione S-transferase (GST; ∼25 kDa) with GST and glutathione peroxidase (GPx) activities. Exposing cells to cobalt chloride also caused the moderate upregulation of NCI16 and PcGST1 mRNAs. Both nickel sulfate and cobalt chloride dose dependently induced NCI16 and PcGST1 mRNAs, but with different profiles. Nickel treatment caused a continuous increase in PcGST1 and NCI16 mRNA levels for up to 3 and 6 days, respectively, and a notable increase in H2O2 concentrations in P. caudatum. NCI16 expression was significantly enhanced by incubating cells with H2O2, implying that NCI16 induction in the presence of nickel ions is caused by reactive oxygen species (ROS). On the other hand, PcGST1 was highly induced by the antioxidant tert-butylhydroquinone (tBHQ) but not by H2O2, suggesting that different mechanisms mediate the induction of NCI16 and PcGST1. We introduced a luciferase reporter vector with an ∼0.42-kb putative PcGST1 promoter into cells and then exposed the transformants to nickel sulfate. This resulted in significant luciferase upregulation, indicating that the putative PcGST1 promoter contains a nickel-responsive element. Our nickel-inducible system also may be applicable to the efficient expression of proteins that are toxic to host cells or require temporal control. PMID:25001407

  9. "Candidatus Gortzia shahrazadis", a Novel Endosymbiont of Paramecium multimicronucleatum and a Revision of the Biogeographical Distribution of Holospora-Like Bacteria.

    PubMed

    Serra, Valentina; Fokin, Sergei I; Castelli, Michele; Basuri, Charan K; Nitla, Venkatamahesh; Verni, Franco; Sandeep, Bhagavatula V; Kalavati, Chaganti; Petroni, Giulio

    2016-01-01

    Holospora spp. and "Candidatus Gortzia infectiva", known as Holospora-like bacteria (HLB), are commonly found as nuclear endosymbionts of ciliates, especially the Paramecium genus. HLB are related by phylogenetic relationships, morphological features, and life-cycles, which involve two alternating morphotypes: reproductive and infectious forms (RF, IF). In this paper we describe a novel species belonging to the "Ca. Gortzia" genus, detected in P. multimicronucleatum, a ciliate for which infection by an HLB has not been reported, discovered in India. This novel endosymbiont shows unusual and surprising features with respect to other HLB, such as large variations in IF morphology and the occasional ability to reproduce in the host cytoplasm. We propose the name of "Candidatus Gortzia shahrazadis" for this novel HLB. Moreover, we report two additional species of HLB from Indian Paramecium populations: "Ca. Gortzia infectiva" (from P. jenningsi), and H. obtusa (from P. caudatum); the latter is the first record of Holospora from a tropical country. Although tropical, we retrieved H. obtusa at an elevation of 706 m corresponding to a moderate climate not unlike conditions where Holospora are normally found, suggesting the genus Holospora does exist in tropical countries, but restricted to higher elevations.

  10. Bug22p, a Conserved Centrosomal/Ciliary Protein Also Present in Higher Plants, Is Required for an Effective Ciliary Stroke in Paramecium ▿ †

    PubMed Central

    Laligné, C.; Klotz, C.; Garreau de Loubresse, N.; Lemullois, M.; Hori, M.; Laurent, F. X.; Papon, J. F.; Louis, B.; Cohen, J.; Koll, F.

    2010-01-01

    Centrioles, cilia, and flagella are ancestral conserved organelles of eukaryotic cells. Among the proteins identified in the proteomics of ciliary proteins in Paramecium, we focus here on a protein, Bug22p, previously detected by cilia and basal-body high-throughput studies but never analyzed per se. Remarkably, this protein is also present in plants, which lack centrioles and cilia. Bug22p sequence alignments revealed consensus positions that distinguish species with centrioles/cilia from plants. In Paramecium, antibody and green fluorescent protein (GFP) fusion labeling localized Bug22p in basal bodies and cilia, and electron microscopy immunolabeling refined the localization to the terminal plate of the basal bodies, the transition zone, and spots along the axoneme, preferentially between the membrane and the microtubules. RNA interference (RNAi) depletion of Bug22p provoked a strong decrease in swimming speed, followed by cell death after a few days. High-speed video microscopy and morphological analysis of Bug22p-depleted cells showed that the protein plays an important role in the efficiency of ciliary movement by participating in the stroke shape and rigidity of cilia. The defects in cell swimming and growth provoked by RNAi can be complemented by expression of human Bug22p. This is the first reported case of complementation by a human gene in a ciliate. PMID:20118210

  11. Effect of 808 nm Diode Laser on Swimming Behavior, Food Vacuole Formation and Endogenous ATP Production of Paramecium primaurelia (Protozoa).

    PubMed

    Amaroli, Andrea; Ravera, Silvia; Parker, Steven; Panfoli, Isabella; Benedicenti, Alberico; Benedicenti, Stefano

    2015-01-01

    Photobiomodulation (PBM) has been used in clinical practice for more than 40 years. To clarify the mechanisms of action of PBM at cellular and organism levels, we investigated its effect on Paramecium primaurelia (Protozoa) irradiated by an 808 nm infrared diode laser with a flat-top handpiece (1 W in CW). Our results led to the conclusion that: (1) the 808 nm laser stimulates the P. primaurelia without a thermal effect, (2) the laser effect is demonstrated by an increase in swimming speed and in food vacuole formation, (3) the laser treatment affects endogenous adenosine triphosphate (ATP) production in a positive way, (4) the effects of irradiation dose suggest an optimum exposure time of 50 s (64 J cm(-2) of fluence) to stimulate the Paramecium cells; irradiation of 25 s shows no effect or only mild effects and irradiation up to 100 s does not increase the effect observed with 50 s of treatment, (5) the increment of endogenous ATP concentration highlights the positive photobiomodulating effect of the 808 nm laser and the optimal irradiation conditions by the flat-top handpiece. © 2015 The American Society of Photobiology.

  12. Paramecium tetraurelia chromatin assembly factor-1-like protein PtCAF-1 is involved in RNA-mediated control of DNA elimination.

    PubMed

    Ignarski, Michael; Singh, Aditi; Swart, Estienne C; Arambasic, Miroslav; Sandoval, Pamela Y; Nowacki, Mariusz

    2014-10-29

    Genome-wide DNA remodelling in the ciliate Paramecium is ensured by RNA-mediated trans-nuclear crosstalk between the germline and the somatic genomes during sexual development. The rearrangements include elimination of transposable elements, minisatellites and tens of thousands non-coding elements called internally eliminated sequences (IESs). The trans-nuclear genome comparison process employs a distinct class of germline small RNAs (scnRNAs) that are compared against the parental somatic genome to select the germline-specific subset of scnRNAs that subsequently target DNA elimination in the progeny genome. Only a handful of proteins involved in this process have been identified so far and the mechanism of DNA targeting is unknown. Here we describe chromatin assembly factor-1-like protein (PtCAF-1), which we show is required for the survival of sexual progeny and localizes first in the parental and later in the newly developing macronucleus. Gene silencing shows that PtCAF-1 is required for the elimination of transposable elements and a subset of IESs. PTCAF-1 depletion also impairs the selection of germline-specific scnRNAs during development. We identify specific histone modifications appearing during Paramecium development which are strongly reduced in PTCAF-1 depleted cells. Our results demonstrate the importance of PtCAF-1 for the epigenetic trans-nuclear cross-talk mechanism. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  13. An ion-current mutant of Paramecium tetraurelia with defects in the primary structure and post-translational N-methylation of calmodulin

    SciTech Connect

    Wallen-Friedman, M.A.

    1988-01-01

    My work on pantophobiac A{sup 2} (pntA{sup 2}), a behavioral mutant of Paramecium tetraurelia, suggest that the Ca{sup ++}-binding protein calmodulin (CaM), and post-translation N-methylation of CaM, are important for Ca{sup ++}-related ion-current function. Calmodulin from wild-type Paramecium has two sites of lysine-N-methylation. Both of these sites are almost fully methylated in vivo; thus wild-type calmodulin is a poor substrate for N-methylation in vitro. In contrast, pntA/{sup 2} CaM can be heavily N-methylated in vitro, suggesting that the mutant calmodulin is under-methylated in vivo. Amino-acid composition analysis showed that CaM lysine 115 is undermethylated in pntA{sup 2}. Once pntA{sup 2} CaM is N-methylated, the (methyl-{sup 3}H) group does not turn over in either wild-type or pntA{sup 2} cytoplasmic fractions. The methylating enzymes in pntA{sup 2} high-speed supernatant fractions are active, but may be less robust than those of the wild type, suggesting a possible control of these enzymes by CaM.

  14. Bug22p, a conserved centrosomal/ciliary protein also present in higher plants, is required for an effective ciliary stroke in Paramecium.

    PubMed

    Laligné, C; Klotz, C; de Loubresse, N Garreau; Lemullois, M; Hori, M; Laurent, F X; Papon, J F; Louis, B; Cohen, J; Koll, F

    2010-04-01

    Centrioles, cilia, and flagella are ancestral conserved organelles of eukaryotic cells. Among the proteins identified in the proteomics of ciliary proteins in Paramecium, we focus here on a protein, Bug22p, previously detected by cilia and basal-body high-throughput studies but never analyzed per se. Remarkably, this protein is also present in plants, which lack centrioles and cilia. Bug22p sequence alignments revealed consensus positions that distinguish species with centrioles/cilia from plants. In Paramecium, antibody and green fluorescent protein (GFP) fusion labeling localized Bug22p in basal bodies and cilia, and electron microscopy immunolabeling refined the localization to the terminal plate of the basal bodies, the transition zone, and spots along the axoneme, preferentially between the membrane and the microtubules. RNA interference (RNAi) depletion of Bug22p provoked a strong decrease in swimming speed, followed by cell death after a few days. High-speed video microscopy and morphological analysis of Bug22p-depleted cells showed that the protein plays an important role in the efficiency of ciliary movement by participating in the stroke shape and rigidity of cilia. The defects in cell swimming and growth provoked by RNAi can be complemented by expression of human Bug22p. This is the first reported case of complementation by a human gene in a ciliate.

  15. Paramecium putrinum (Ciliophora, Protozoa): the first insight into the variation of two DNA fragments - molecular support for the existence of cryptic species.

    PubMed

    Tarcz, Sebastian; Rautian, Maria; Potekhin, Alexey; Sawka, Natalia; Beliavskaya, Alexandra; Kiselev, Andrey; Nekrasova, Irina; Przyboś, Ewa

    2014-04-01

    Paramecium putrinum (Claparede & Lachmann 1858) is one of the smallest (80-140 μm long) species of the genus Paramecium. Although it commonly occurs in freshwater reservoirs, no molecular studies of P. putrinum have been conducted to date. Herein we present an assessment of molecular variation in 27 strains collected from widely separated populations by using two selected DNA fragments (ITS1-5.8S-ITS2-5'LSU rDNA and COI mtDNA). Both the trees and haplotype networks reconstructed for both genome fragments show that the studied strains of P. putrinum form five main haplogroups. The mean distance between the studied strains is p-distance=0.007/0.068 (rDNA/COI) and exhibits similar variability as that between P. bursaria syngens. Based on these data, one could hypothesize that the clusters revealed in the present study may correspond to previously reported syngens and that there are at least five cryptic species within P. putrinum. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. “Candidatus Gortzia shahrazadis”, a Novel Endosymbiont of Paramecium multimicronucleatum and a Revision of the Biogeographical Distribution of Holospora-Like Bacteria

    PubMed Central

    Serra, Valentina; Fokin, Sergei I.; Castelli, Michele; Basuri, Charan K.; Nitla, Venkatamahesh; Verni, Franco; Sandeep, Bhagavatula V.; Kalavati, Chaganti; Petroni, Giulio

    2016-01-01

    Holospora spp. and “Candidatus Gortzia infectiva”, known as Holospora-like bacteria (HLB), are commonly found as nuclear endosymbionts of ciliates, especially the Paramecium genus. HLB are related by phylogenetic relationships, morphological features, and life-cycles, which involve two alternating morphotypes: reproductive and infectious forms (RF, IF). In this paper we describe a novel species belonging to the “Ca. Gortzia” genus, detected in P. multimicronucleatum, a ciliate for which infection by an HLB has not been reported, discovered in India. This novel endosymbiont shows unusual and surprising features with respect to other HLB, such as large variations in IF morphology and the occasional ability to reproduce in the host cytoplasm. We propose the name of “Candidatus Gortzia shahrazadis” for this novel HLB. Moreover, we report two additional species of HLB from Indian Paramecium populations: “Ca. Gortzia infectiva” (from P. jenningsi), and H. obtusa (from P. caudatum); the latter is the first record of Holospora from a tropical country. Although tropical, we retrieved H. obtusa at an elevation of 706 m corresponding to a moderate climate not unlike conditions where Holospora are normally found, suggesting the genus Holospora does exist in tropical countries, but restricted to higher elevations. PMID:27867371

  17. The protozoan, Paramecium primaurelia, as a non-sentient model to test laser light irradiation: The effects of an 808nm infrared laser diode on cellular respiration.

    PubMed

    Amaroli, Andrea; Ravera, Silvia; Parker, Steven; Panfoli, Isabella; Benedicenti, Alberico; Benedicenti, Stefano

    2015-07-01

    Photobiomodulation (PBM) has been used in clinical practice for more than 40 years. Unfortunately, conflicting literature has led to the labelling of PBM as a complementary or alternative medicine approach. However, past and ongoing clinical and research studies by reputable investigators have re-established the merits of PBM as a genuine medical therapy, and the technique has, in the last decade, seen an exponential increase in the numbers of clinical instruments available, and their applications. This resurgence has led to a clear need for appropriate experimental models to test the burgeoning laser technology being developed for medical applications. In this context, an ethical model that employs the protozoan, Paramecium primaurelia, is proposed. We studied the possibility of using the measure of oxygen consumption to test PBM by irradiation with an infrared or near-infrared laser. The results show that an 808nm infrared laser diode (1W; 64J/cm²) affects cellular respiration in P. primaurelia, inducing, in the irradiated cells, a significantly (p < 0.05) increased oxygen consumption of about 40%. Our findings indicate that Paramecium can be an excellent tool in biological assays involving infrared and near-infrared PBM, as it combines the advantages of in vivo results with the practicality of in vitro testing. This test represents a fast, inexpensive and straightforward assay, which offers an alternative to both traditional in vivo testing and more expensive mammalian cellular cultures. 2015 FRAME.

  18. Isolation and characterization of a virus (CvV-BW1) that infects symbiotic algae of Paramecium bursaria in Lake Biwa, Japan

    PubMed Central

    2010-01-01

    Background We performed an environmental study of viruses infecting the symbiotic single-celled algae of Paramecium bursaria (Paramecium bursaria Chlorella virus, PBCV) in Lake Biwa, the largest lake in Japan. The viruses detected were all Chlorella variabilis virus (CvV = NC64A virus). One of them, designated CvV-BW1, was subjected to further characterization. Results CvV-BW1 formed small plaques and had a linear DNA genome of 370 kb, as judged by pulsed-field gel electrophoresis. Restriction analysis indicated that CvV-BW1 DNA belongs to group H, one of the most resistant groups among CvV DNAs. Based on a phylogenetic tree constructed using the dnapol gene, CvV was classified into two clades, A and B. CvV-BW1 belonged to clade B, in contrast to all previously identified virus strains of group H that belonged to clade A. Conclusions We conclude that CvV-BW1 composes a distinct species within C. variabilis virus. PMID:20831832

  19. Distinct RNA-dependent RNA polymerases are required for RNAi triggered by double-stranded RNA versus truncated transgenes in Paramecium tetraurelia

    PubMed Central

    Marker, Simone; Le Mouël, Anne; Meyer, Eric; Simon, Martin

    2010-01-01

    In many eukaryotes, RNA-dependent RNA polymerases (RdRPs) play key roles in the RNAi pathway. They have been implicated in the recognition and processing of aberrant transcripts triggering the process, and in amplification of the silencing response. We have tested the functions of RdRP genes from the ciliate Paramecium tetraurelia in experimentally induced and endogenous mechanisms of gene silencing. In this organism, RNAi can be triggered either by high-copy, truncated transgenes or by directly feeding cells with double-stranded RNA (dsRNA). Surprisingly, dsRNA-induced silencing depends on the putatively functional RDR1 and RDR2 genes, which are required for the accumulation of both primary siRNAs and a distinct class of small RNAs suggestive of secondary siRNAs. In contrast, a third gene with a highly divergent catalytic domain, RDR3, is required for siRNA accumulation when RNAi is triggered by truncated transgenes. Our data further implicate RDR3 in the accumulation of previously described endogenous siRNAs and in the regulation of the surface antigen gene family. While only one of these genes is normally expressed in any clonal cell line, the knockdown of RDR3 leads to co-expression of multiple antigens. These results provide evidence for a functional specialization of Paramecium RdRP genes in distinct RNAi pathways operating during vegetative growth. PMID:20200046

  20. Paramecium tetraurelia chromatin assembly factor-1-like protein PtCAF-1 is involved in RNA-mediated control of DNA elimination

    PubMed Central

    Ignarski, Michael; Singh, Aditi; Swart, Estienne C.; Arambasic, Miroslav; Sandoval, Pamela Y.; Nowacki, Mariusz

    2014-01-01

    Genome-wide DNA remodelling in the ciliate Paramecium is ensured by RNA-mediated trans-nuclear crosstalk between the germline and the somatic genomes during sexual development. The rearrangements include elimination of transposable elements, minisatellites and tens of thousands non-coding elements called internally eliminated sequences (IESs). The trans-nuclear genome comparison process employs a distinct class of germline small RNAs (scnRNAs) that are compared against the parental somatic genome to select the germline-specific subset of scnRNAs that subsequently target DNA elimination in the progeny genome. Only a handful of proteins involved in this process have been identified so far and the mechanism of DNA targeting is unknown. Here we describe chromatin assembly factor-1-like protein (PtCAF-1), which we show is required for the survival of sexual progeny and localizes first in the parental and later in the newly developing macronucleus. Gene silencing shows that PtCAF-1 is required for the elimination of transposable elements and a subset of IESs. PTCAF-1 depletion also impairs the selection of germline-specific scnRNAs during development. We identify specific histone modifications appearing during Paramecium development which are strongly reduced in PTCAF-1 depleted cells. Our results demonstrate the importance of PtCAF-1 for the epigenetic trans-nuclear cross-talk mechanism. PMID:25270876

  1. The ionic composition of the contractile vacuole fluid of Paramecium mirrors ion transport across the plasma membrane.

    PubMed

    Stock, Christian; Grønlien, Heidi K; Allen, Richard D

    2002-09-01

    In vivo K+, Na+, Ca2+, Cl- and H+ activities in the cytosol and the contractile vacuole fluid, the overall cytosolic osmolarity, the fluid segregation rate per contractile vacuole and the membrane potential of the contractile vacuole complex of Paramecium multimicronucleatum were determined in cells adapted to 24 or 124 mosm l(-1) solutions containing as the monovalent cation(s): 1) 2 mmol l(-1) K+; 2) 2 mmol l(-1) Na+; 3) 1 mmol l(-1) K+ plus 1 mmol l(-1) Na+; or 4) 2 mmol l(-1) choline. In cells adapted to a given external osmolarity i) the fluid segregation rate was the same if adapted to either K+ or Na+, twice as high when adapted to solutions containing both K+ and Na+, and reduced by 50% or more in solutions containing only choline, ii) the fluid of the contractile vacuole was always hypertonic to the cytosol while the sum of the ionic activities measured in the fluid of the contractile vacuole was the same in cells adapted to either K+ or Na+, at least 25% higher in cells adapted to solutions containing both K+ and Na+, and was reduced by 55% or more in solutions containing only choline, iii) the cytosolic osmolarity was the same in cells adapted to K+ alone, to Na+ alone or to both K+ and Na+, whereas it was significantly lower in cells adapted to choline. At a given external osmolarity, a positive relationship between the osmotic gradient across the membrane of the contractile vacuole complex and the fluid segregation rate was observed. We conclude that both the plasma membrane and the membrane of the contractile vacuole complex play roles in fluid segregation. The presence of external Na+ moderated K+ uptake and caused the Ca2+ activity in the contractile vacuole fluid to rise dramatically. Thus, Ca2+ can be eliminated through the contractile vacuole complex when Na+ is present externally. The membrane potential of the contractile vacuole complex remained essentially the same regardless of the external ionic conditions and the ionic composition of the

  2. [Macronuclear DNA and total protein contents of mating types I and II of Paramecium primaurelia, during the phase of maturity and the transition to senescence. Preliminary observations].

    PubMed

    Delmonte Corrado, M U; Crippa Franceschi, T

    1992-01-01

    Concerning the studies on mating type differentiation and life cycle development in Paramecium primaurelia stock 90, both macronuclear DNA and total protein contents have been measured cytofluorometrically in mating type I and mating type II isogenic cell lines growing in logarithmic phase, throughout their maturity period and transition to senescence. The target was to investigate whether the two mating types undergo clonal decline in different times, as the previous studies suggested. The results indicate that, throughout the maturity period, macronuclear DNA and total protein contents vary both in mating type I and mating type II cell lines; moreover, aged phenotypes as the dramatic decrease of both contents, firstly occur in mating type II which, therefore, appears to be submitted to clonal decline before mating type I.

  3. Immunological studies of isolated particulates of Paramecium aurella. I. Antigenic relationships between cytoplasmic organelles and evidence for mitochondrial variations as demonstrated by gel diffusion.

    PubMed

    FINGER, I; KABACK, M; KITTNER, P; HELLER, C

    1960-12-01

    Mitochondria and other particulates-cilia, trichocysts, and "small granules"-have been isolated from several stocks of Paramecium aurelia, syngen 2. Antisera against these particles and against breis have been used to characterize the fractions by diffusion in gel. Evidence is presented for the relationship of particles, as demonstrated by immunologic cross-reactivity of the soluble antigens extracted from them. Although some antigens are unique for a fraction, cross-reacting antigens in two or more fractions, as determined by "spur" formation in agar, suggest a relationship between morphologically diverse particles. A procedure for studying cross-reactions in gels is described using the specific immobilization antigens as a model. The localization of these antigens within cilia, and perhaps trichocysts, has been confirmed. Other organelles, specifically mitochondria and "small granules," appear to alter their specificity spontaneously and reversibly during cell reproduction, a pattern reminiscent of the immobilization serotypes which can transform to one another during clonal growth.

  4. Microsurgical analysis of the clonal age and the cell-cycle stage required for the onset of autogamy in Paramecium tetraurelia.

    PubMed

    Mikami, K; Koizumi, S

    1983-11-01

    When autogamy was induced in competent cells of Paramecium tetraurelia by depriving them of food, the onset of autogamy was preceded by a critical fission which occurred in the starvation medium. When the cells were fed again immediately after the fission, they did not undergo autogamy. However, they did undergo autogamy when they were fed later than 1 hr after the critical fission. The irreversible differentiation for autogamy seems to be at about 1 hr after the critical fission. This procedure thus provides the opportunity to induce autogamy synchronously. The result of macronuclear transplantation demonstrated that autogamy was under the control of macronucleus. Moreover, the clonal age required for autogamy was found to be shortened by repetitive elimination of a part of the macronucleus. The result can be explained by the hypothesis that clonal age is measured in rounds of chromosome replication or DNA synthesis rather than cell divisions.

  5. Molecular aspects of calcium signalling at the crossroads of unikont and bikont eukaryote evolution--the ciliated protozoan Paramecium in focus.

    PubMed

    Plattner, Helmut

    2015-03-01

    The ciliated protozoan, Paramecium tetraurelia has a high basic Ca(2+) leakage rate which is counteracted mainly by export through a contractile vacuole complex, based on its V-type H(+)-ATPase activity. In addition Paramecium cells dispose of P-type Ca(2+)-ATPases, i.e. a plasmamembrane and a sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase (PMCA, SERCA). Antiporter systems are to be expected, as inferred from indirect evidence. Among the best known cytosolic Ca(2+)-binding proteins, calmodulin activates Ca(2+) influx channels in the somatic cell membrane, but inactivates Ca(2+) influx channels in cilia, where it, thus, ends ciliary reversal induced by depolarization via channels in the somatic cell membrane. Centrin inactivates Ca(2+) signals after stimulation by its high capacity/low affinity binding sites, whereas its high affinity sites regulate some other functions. Cortical Ca(2+) stores (alveolar sacs) are activated during stimulated trichocyst exocytosis and thereby mediate store-operated Ca(2+) entry (SOCE). Ca(2+) release channels (CRCs) localised to alveoli and underlying SOCE are considered as Ryanodine receptor-like proteins (RyR-LPs) which are members of a CRC family with 6 subfamilies. These also encompass genuine inositol 1,4,5-trisphosphate receptors (IP3Rs) and intermediates between the two channel types. All IP3R/RyR-type CRCs possess six carboxyterminal transmembrane domains (TMD), with a pore domain between TMD 5 and 6, endowed with a characteristic selectivity filter. There are reasons to assume a common ancestor molecule for such channels and diversification further on in evolution. The distinct distribution of specific CRCs in the different vesicles undergoing intracellular trafficking suggests constitutive formation of very locally restricted Ca(2+) signals during vesicle-vesicle interaction. In summary, essential steps of Ca(2+) signalling already occur at this level of evolution, including an unexpected multitude of CRCs. For dis

  6. Cell surface complexes ('cortices') isolated from Paramecium tetraurelia cells as a model system for analysing exocytosis in vitro in conjunction with microinjection studies.

    PubMed Central

    Lumpert, C J; Kersken, H; Plattner, H

    1990-01-01

    Cortex preparations isolated from Paramecium tetraurelia cells consist of surface with secretory organelles (trichocysts) still attached. In the absence of nucleotides, in media with a pCa of 5-5.5 and a pH of greater than or equal to 6.5, maximal exocytosis occurred when the Mg2+ concentration was lowered from 10 to 0.5 mM. ATP, as well as its non-hydrolysable analogues adenosine 5'-[gamma-thio]triphosphate (ATP[S]) and adenosine 5'[beta gamma-imido]triphosphate (App[NH]p), inhibited exocytosis at a concentration equivalent to that occurring in vivo (as determined by h.p.l.c.), but preincubation with ATP augmented the exocytotic response. GTP and its analogues only slightly stimulated exocytosis in vitro, but sensitivity to Ca2+ was increased significantly, in particular with GTP. These effects of nucleotides were rapidly reversible. Intracellular GTP concentrations (0.35 mM) would suffice for full activation with the pCai values assumed to occur in these cells during activation. On microinjection, ATP inhibited the secretagogue response in intact cells. Whereas microinjected GTP stimulated exocytosis (membrane fusion) without a secretagogue added, Gpp[NH]p remained without any effect; GTP[S] permanently abolished any triggered secretory response. Concomitantly, h.p.l.c. analysis of triggered and untriggered cells showed that GTP hydrolysis occurs immediately after synchronous (1 s) exocytosis in vivo. The precise site(s) of action of GTP during signal transduction in Paramecium cells remain to be determined. Images Fig. 4. PMID:2390058

  7. Genetic analysis of membrane differentiation in Paramecium. Freeze- fracture study of the trichocyst cycle in wild-type and mutant strains

    PubMed Central

    1976-01-01

    Using a series of mutants of Paramecium tetraurelia, we demonstrate, for the first time, changes in the internal structure of the cell membrane, as revealed by freeze-fracture, that correspond to specific single gene mutations. On the plasma membrane of Paramecium circular arrays of particles mark the sites of attachment of the tips of the intracellular secretory organelles-trichocysts. In wild-type paramecia, where attached trichocysts can be expelled by exocytosis under various stimuli, the plasma membrane array is composed of a double outer ring of particles (300 nm in diameter) and inside the ring a central rosette (fusion rosette) of particles (76 nm in diameter). Mutant nd9, characterized by a thermosensitive ability to discharge trichocysts, shows the same organization in cells grown at the permissive temperature (18 degrees C), while in cells grown at the nonpermissive temperature (27 degrees C) the rosette is missing. In mutant tam 8, characterized by normal but unattached trichocysts, and in mutant tl, completely devoid of trichocysts, no rosette is formed and the outer rings always show a modified configuration called "parentheses", also found in wild-type and in nd9 (18 degrees C) cells. From this comparison between wild type and mutants, we conclude: (a) that the formation of parentheses is a primary differentiation of the plasma membrane, independent of the presence of trichocysts, while the secondary transformation of parentheses into circular arrays and the formation of the rosette are triggered by interaction between trichocysts and plasma membranes; and (b) that the formation of the rosette is a prerequisite for trichocyst exocytosis. PMID:1254639

  8. Functional and fluorochrome analysis of an exocytotic mutant yields evidence of store-operated Ca2+ influx in Paramecium.

    PubMed

    Mohamed, I; Klauke, N; Hentschel, J; Cohen, J; Plattner, H

    2002-05-01

    A non-discharge mutant of Paramecium tetraurelia (nd12-35 degrees C, lacking exocytotic response upon stimulation with the nonpermeable polycationic secretagogue aminoethyldextran, AED), in the pawnA genetic context (d4-500r, lacking ciliary voltage-dependent Ca2+ influx), was shown to lack (45)Ca2+ entry from outside upon AED stimulation. In contrast, cells grown at 25 degrees C behave like the wildtype. To check the functional properties in more detail, fluorochrome-loaded 35 degrees C cells were stimulated, not only with AED (EC(100) = 10(-6) M in wildtype cells), but also with 4-chloro-meta-cresol, (4CmC, 0.5 mM), a permeable activator of ryanodine receptor-type Ca2+ release channels, usually at extracellular [Ca2+] of 50 microM, and eventually with a Ca2+ chelator added. We confirm that pwA-nd12(35 degrees C) cells lack any Ca2+ influx and any exocytosis of trichocysts in response to any stimulus. As we determined by x-ray microanalysis, total calcium content in alveolar sacs (subplasmalemmal stores) known to be mobilized upon exocytosis stimulation in wild-type cells, contain about the same total calcium in 35 degrees C as in 25 degrees C cells, and Ca2+ mobilization from alveoli by AED or 4CmC is also nearly the same. Due to the absence of any AED-induced Ca2+ influx in 35 degrees C cells and normal Ca2+ release from stores found by x-ray microanalysis one can exclude a "CICR"-type mechanism (Ca2+-induced Ca2+ release) and imply that normally a store-operated Ca2+ ("SOC") influx would occur (as in 25 degrees C cells). Furthermore, 35 degrees C cells display a significantly lower basal intracellular [Ca2+], so that any increase upon stimulation may be less expressed or even remain undetected. Under these conditions, any mobilization of Ca2+ from stores cannot compensate for the lack of Ca2+ influx, particularly since normally both components have to cooperate to achieve full exocytotic response. Also striking is our finding that 35 degrees C cells are unable

  9. Calcium signalling in the ciliated protozoan model, Paramecium: strict signal localisation by epigenetically controlled positioning of different Ca²⁺-channels.

    PubMed

    Plattner, Helmut

    2015-03-01

    The Paramecium tetraurelia cell is highly organised, with regularly spaced elements pertinent to Ca(2+) signalling under epigenetic control. Vesicles serving as stationary Ca(2+) stores or undergoing trafficking contain Ca(2+)-release channels (PtCRCs) which, according to sequence and domain comparison, are related either to inositol 1,4,5-trisphosphate (InsP3) receptors (IP3R) or to ryanodine receptor-like proteins (RyR-LP) or to both, with intermediate characteristics or deviation from conventional domain structure. Six groups of such PtCRCs have been found. The ryanodine-InsP3-receptor homology (RIH) domain is not always recognisable, in contrast to the channel domain with six trans-membrane domains and the pore between transmembrane domain 5 and 6. Two CRC subtypes tested more closely, PtCRC-II and PtCRC-IV, with and without an InsP3-binding domain, reacted to InsP3 and to caffeine, respectively, and hence represent IP3Rs and RyR-LPs. IP3Rs occur in the contractile vacuole complex where they allow for stochastic constitutive Ca(2+) reflux into the cytosol. RyR-LPs are localised to cortical Ca(2+) stores; they are engaged in dense core-secretory vesicle exocytosis by Ca(2+) release, superimposed by Ca(2+)-influx via non-ciliary Ca(2+)-channels. One or two different types of PtCRCs also occur in other vesicles undergoing trafficking. Since the PtCRCs described combine different features they are considered derivatives of primitive precursors. The highly regular, epigenetically controlled design of a Paramecium cell allows it to make Ca(2+) available very locally, in a most efficient way, along predetermined trafficking pathways, including regulation of exocytosis, endocytosis, phagocytosis and recycling phenomena. The activity of cilia is also regulated by Ca(2+), yet independently from any CRCs, by de- and hyperpolarisation of the cell membrane potential. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Disentangling the Taxonomy of Rickettsiales and Description of Two Novel Symbionts ("Candidatus Bealeia paramacronuclearis" and "Candidatus Fokinia cryptica") Sharing the Cytoplasm of the Ciliate Protist Paramecium biaurelia.

    PubMed

    Szokoli, Franziska; Castelli, Michele; Sabaneyeva, Elena; Schrallhammer, Martina; Krenek, Sascha; Doak, Thomas G; Berendonk, Thomas U; Petroni, Giulio

    2016-12-15

    In the past 10 years, the number of endosymbionts described within the bacterial order Rickettsiales has constantly grown. Since 2006, 18 novel Rickettsiales genera inhabiting protists, such as ciliates and amoebae, have been described. In this work, we characterize two novel bacterial endosymbionts from Paramecium collected near Bloomington, IN. Both endosymbiotic species inhabit the cytoplasm of the same host. The Gram-negative bacterium "Candidatus Bealeia paramacronuclearis" occurs in clumps and is frequently associated with the host macronucleus. With its electron-dense cytoplasm and a distinct halo surrounding the cell, it is easily distinguishable from the second smaller symbiont, "Candidatus Fokinia cryptica," whose cytoplasm is electron lucid, lacks a halo, and is always surrounded by a symbiontophorous vacuole. For molecular characterization, the small-subunit rRNA genes were sequenced and used for taxonomic assignment as well as the design of species-specific oligonucleotide probes. Phylogenetic analyses revealed that "Candidatus Bealeia paramacronuclearis" clusters with the so-called "basal" Rickettsiales, and "Candidatus Fokinia cryptica" belongs to "Candidatus Midichloriaceae." We obtained tree topologies showing a separation of Rickettsiales into at least two groups: one represented by the families Rickettsiaceae, Anaplasmataceae, and "Candidatus Midichloriaceae" (RAM clade), and the other represented by "basal Rickettsiales," including "Candidatus Bealeia paramacronuclearis." Therefore, and in accordance with recent publications, we propose to limit the order Rickettsiales to the RAM clade and to raise "basal Rickettsiales" to an independent order, Holosporales ord. nov., inside Alphaproteobacteria, which presently includes four family-level clades. Additionally, we define the family "Candidatus Hepatincolaceae" and redefine the family Holosporaceae IMPORTANCE: In this paper, we provide the characterization of two novel bacterial symbionts

  11. Molecular characterization of a sarco(endo)plasmic reticulum Ca2+-ATPase gene from Paramecium tetraurelia and localization of its gene product to sub-plasmalemmal calcium stores.

    PubMed Central

    Hauser, K; Pavlovic, N; Kissmehl, R; Plattner, H

    1998-01-01

    A cDNA encoding the gene for a sarco(endo)plasmic reticulum-type Ca2+-ATPase (SERCA) was isolated from a cDNA library of Paramecium tetraurelia by using degenerated primers according to conserved domains of SERCA-type ATPases. The identified nucleotide sequence (PtSERCA) is 3114 nucleotides in length with an open reading frame of 1037 amino acids. An intron of only 22 nucleotides occurs. Homology searches for the deduced amino acid sequence revealed 38-49% similarity to SERCA-type ATPases from organisms ranging from protozoans to mammals, with no more similarity to some parasitic protozoa of the same phylum. The calculated molecular mass of the encoded protein is 114.7 kDa. It contains the typical 10 transmembrane domains of SERCA-type ATPases and other conserved domains, such as the phosphorylation site and the ATP binding site. However, there are no binding sites for phospholamban and thapsigargin present in the PtSERCA. Antibodies raised against a cytoplasmic loop peptide between the phosphorylation site and the ATP binding site recognize on Western blots a protein of 106 kDa, exclusively in the fraction of sub-plasmalemmal calcium stores ('alveolar sacs'). In immunofluorescence studies the antibodies show labelling exclusively in the cell cortex of permeabilized cells in a pattern characteristic of the arrangement of alveolar sacs. When alveolar sacs where tested for phosphoenzyme-intermediate formation a phosphoprotein of the same molecular mass (106 kDa) could be identified. PMID:9693098

  12. Endosymbiosis of Chlorella species to the ciliate Paramecium bursaria alters the distribution of the host's trichocysts beneath the host cell cortex.

    PubMed

    Kodama, Yuuki; Fujishima, Masahiro

    2011-04-01

    Each symbiotic Chlorella of the ciliate Paramecium bursaria is enclosed in a perialgal vacuole membrane derived from the host digestive vacuole membrane. Alga-free paramecia and symbiotic algae can grow independently. Mixing them experimentally can cause reinfection. Earlier, we reported that the symbiotic algae appear to push the host trichocysts aside to become fixed beneath the host cell cortex during the algal reinfection process. Indirect immunofluorescence microscopy with a monoclonal antibody against the trichocysts demonstrates that the trichocysts change their locality to form algal attachment sites and decrease their density beneath the host cell cortex through algal reinfection. Transmission electron microscopy to detect acid phosphatase activity showed that some trichocysts near the host cell cortex are digested by the host lysosomal fusion during algal reinfection. Removal of algae from the host cell using cycloheximide recovers the trichocyst's arrangement and number near the host cell cortex. These results indicate that symbiotic algae compete for their attachment sites with preexisting trichocysts and that the algae have the ability to ensure algal attachment sites beneath the host cell cortex.

  13. Genomic Characterization of Variable Surface Antigens Reveals a Telomere Position Effect as a Prerequisite for RNA Interference-Mediated Silencing in Paramecium tetraurelia

    PubMed Central

    Baranasic, Damir; Oppermann, Timo; Cheaib, Miriam; Cullum, John; Schmidt, Helmut

    2014-01-01

    ABSTRACT Antigenic or phenotypic variation is a widespread phenomenon of expression of variable surface protein coats on eukaryotic microbes. To clarify the mechanism behind mutually exclusive gene expression, we characterized the genetic properties of the surface antigen multigene family in the ciliate Paramecium tetraurelia and the epigenetic factors controlling expression and silencing. Genome analysis indicated that the multigene family consists of intrachromosomal and subtelomeric genes; both classes apparently derive from different gene duplication events: whole-genome and intrachromosomal duplication. Expression analysis provides evidence for telomere position effects, because only subtelomeric genes follow mutually exclusive transcription. Microarray analysis of cultures deficient in Rdr3, an RNA-dependent RNA polymerase, in comparison to serotype-pure wild-type cultures, shows cotranscription of a subset of subtelomeric genes, indicating that the telomere position effect is due to a selective occurrence of Rdr3-mediated silencing in subtelomeric regions. We present a model of surface antigen evolution by intrachromosomal gene duplication involving the maintenance of positive selection of structurally relevant regions. Further analysis of chromosome heterogeneity shows that alternative telomere addition regions clearly affect transcription of closely related genes. Consequently, chromosome fragmentation appears to be of crucial importance for surface antigen expression and evolution. Our data suggest that RNAi-mediated control of this genetic network by trans-acting RNAs allows rapid epigenetic adaptation by phenotypic variation in combination with long-term genetic adaptation by Darwinian evolution of antigen genes. PMID:25389173

  14. "Candidatus Sonnebornia yantaiensis", a member of candidate division OD1, as intracellular bacteria of the ciliated protist Paramecium bursaria (Ciliophora, Oligohymenophorea).

    PubMed

    Gong, Jun; Qing, Yao; Guo, Xiaohong; Warren, Alan

    2014-02-01

    An intracellular bacterium was discovered in an isolate of Paramecium bursaria from a freshwater pond in Yantai, China. The bacteria were abundant and exclusively found in the cytoplasm of the host which, along with the green alga Chlorella, formed a three-partner consortium that could survive in pure water for at least one week. Cloning, sequencing and phylogenetic analysis of the bacterial 16S rRNA gene showed that the bacterium belonged to the uncultured candidate division OD1, which usually forms part of the rare biosphere. Transmission electron microscopy and fluorescence in situ hybridization (FISH) with specific probes showed that the bacteria were usually located close to the perialgal membranes of endosymbiotic Chlorella cells, and occasionally irregularly distributed throughout the host cytoplasm. The name "Candidatus Sonnebornia yantaiensis" gen. nov., sp. nov. is proposed for the new bacterium. A strongly supported monophyletic subclade, OD1-p, which included the new species, was recognized and this study highlights that protists can be important hosts for rare bacterial taxa. Copyright © 2013 Elsevier GmbH. All rights reserved.

  15. Timing of initiation of macronuclear DNA synthesis is set during the preceding cell cycle in Paramecium tetraurelia: analysis of the effects of abrupt changes in nutrient level

    SciTech Connect

    Ching, A.S.L.; Berger, J.D.

    1986-11-01

    In many eukaryotic organisms, initiation of DNA synthesis is associated with a major control point within the cell cycle and reflects the commitment of the cell to the DNA replication-division portion of the cell cycle. In paramecium, the timing of DNA synthesis initiation is established prior to fission during the preceding cell cycle. DNA synthesis normally starts at 0.25 in the cell cycle. When dividing cells are subjected to abrupt nutrient shift-up by transfer from a chemostat culture to medium with excess food, or shift-down from a well-fed culture to exhausted medium, DNA synthesis initiation in the post-shift cell cycle occurs at 0.25 of the parental cell cycle and not at either 0.25 in the post-shift cell cycle or at 0.25 in the equilibrium cell cycle produced under the post-shift conditions. The long delay prior to initiation of DNA synthesis following nutritional shift-up is not a consequence of continued slow growth because the rate of protein synthesis increases rapidly to the normal level after shift-up. Analysis of the relation between increase in cell mass and initiation of DNA synthesis following nutritional shifts indicates that increase in cell mass, per se, is neither a necessary nor a sufficient condition for initiation of DNA synthesis, in spite of the strong association between accumulation of cell mass and initiation of DNA synthesis in cells growing under steady-state conditions.

  16. Cell division and density of symbiotic Chlorella variabilis of the ciliate Paramecium bursaria is controlled by the host's nutritional conditions during early infection process.

    PubMed

    Kodama, Yuuki; Fujishima, Masahiro

    2012-10-01

    The association of ciliate Paramecium bursaria with symbiotic Chlorella sp. is a mutualistic symbiosis. However, both the alga-free paramecia and symbiotic algae can still grow independently and can be reinfected experimentally by mixing them. Effects of the host's nutritional conditions against the symbiotic algal cell division and density were examined during early reinfection. Transmission electron microscopy revealed that algal cell division starts 24 h after mixing with alga-free P. bursaria, and that the algal mother cell wall is discarded from the perialgal vacuole membrane, which encloses symbiotic alga. Labelling of the mother cell wall with Calcofluor White Stain, a cell-wall-specific fluorochrome, was used to show whether alga had divided or not. Pulse labelling of alga-free P. bursaria cells with Calcofluor White Stain-stained algae with or without food bacteria for P. bursaria revealed that the fluorescence of Calcofluor White Stain in P. bursaria with bacteria disappeared within 3 days after mixing, significantly faster than without bacteria. Similar results were obtained both under constant light and dark conditions. This report is the first describing that the cell division and density of symbiotic algae of P. bursaria are controlled by the host's nutritional conditions during early infection. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  17. Nowa1p and Nowa2p: novel putative RNA binding proteins involved in trans-nuclear crosstalk in Paramecium tetraurelia.

    PubMed

    Nowacki, Mariusz; Zagorski-Ostoja, Wlodzimierz; Meyer, Eric

    2005-09-20

    The germline genome of ciliates is extensively rearranged during development of a new somatic macronucleus from the germline micronucleus, a process that follows sexual events. In Paramecium tetraurelia, single-copy internal eliminated sequences (IESs) and multicopy transposons are eliminated, whereas cellular genes are amplified to approximately 800 n. For a subset of IESs, introduction of the IES sequence into the maternal (prezygotic) macronucleus specifically inhibits excision of the homologous IES in the developing zygotic macronucleus. This and other homology-dependent maternal effects have suggested that rearrangement patterns are epigenetically determined by an RNA-mediated, trans-nuclear comparison, involving the RNA interference pathway, of germline and somatic genomes. We report the identification of novel developmentally regulated RNA binding proteins, Nowa1p and Nowa2p, which are required for the survival of sexual progeny. Green fluorescent protein (GFP) fusions show that Nowa1p accumulates into the maternal macronucleus shortly before meiosis of germline micronuclei and is later transported to developing macronuclei. Nowa1p/2p depletion impairs the elimination of transposons and of those IESs that are controlled by maternal effects, confirming the existence of distinct IES classes. The results indicate that Nowa proteins are essential components of the trans-nuclear-crosstalk mechanism that is responsible for epigenetic programming of genome rearrangements. We discuss implications for the current models of genome scanning in ciliates, a process related to the formation of heterochromatin by RNA interference in other eukaryotes.

  18. ND9P, a novel protein with armadillo-like repeats involved in exocytosis: physiological studies using allelic mutants in paramecium.

    PubMed Central

    Froissard, M; Keller, A M; Cohen, J

    2001-01-01

    In Paramecium, a number of mutants affected in the exocytotic membrane fusion step of the regulated secretory pathway have been obtained. Here, we report the isolation of one of the corresponding genes, ND9, previously suspected to encode a soluble protein interacting with both plasma and trichocyst membranes. Nd9p is a novel polypeptide that contains C-terminal Armadillo-like repeats. Point mutations were found in the first N-terminal quarter of the molecule and in the last putative Armadillo repeat, respectively, for the two thermosensitive mutants, nd9-1 and nd9-2. The different behaviors of these mutants in recovery experiments upon temperature shifts suggest that the N-terminal domain of the molecule may be involved in membrane binding activity, whereas the C-terminal domain is a candidate for protein-protein interactions. The nonsense nd9-3 mutation that produces a short N-terminal peptide has a dominant negative effect on the nd9-1 allele. We show here that, when overexpressed, the dominant negative effect can be produced even on the wild-type allele, suggesting competition for a common target. We suggest that Nd9p could act, like some SNARE proteins, at the membrane-cytosol interface to promote membrane fusion. PMID:11156983

  19. Marked amplification and diversification of products of ras genes from rat brain, Rab GTPases, in the ciliates Tetrahymena thermophila and Paramecium tetraurelia.

    PubMed

    Saito-Nakano, Yumiko; Nakahara, Tohru; Nakano, Kentaro; Nozaki, Tomoyoshi; Numata, Osamu

    2010-01-01

    Small GTPase Rab (products of ras genes from rat brain) is a widely conserved molecular switch among eukaryotes and regulates membrane trafficking pathways. It is generally considered that the number of Rab encoded in the genome correlates with multicellularity; however, we found that unicellular ciliates Tetrahymena thermophila (Tt) and Paramecium tetraurelia (Pt) possess many more Rab genes in their genome than the 64 HsRab genes in the human genome. We succeeded in isolating 86 cDNA clones of 88 TtRab genes in the Tetrahymena genome. By comparing the amino acid sequence of Rab in humans and the budding yeast Saccharomyces cerevisiae, 42 TtRab belonged to subfamilies functionally characterized and designated as conventional Rab, while the remaining 44 TtRab were considered to be species-specific. To examine the diversity of Rab in ciliates, we searched for Rab genes in the genome database of P. tetraurelia. Overall, 229 PtRab genes were found and categorized as 157 conventional and 72 species-specific PtRab, respectively. Among them, nine PtRab genes showed high homology to seven TtRab, suggesting the conservation of ciliate-specific Rab. These data suggested that the range of Rab is markedly amplified and diversified in ciliates, which may support the elaborate cellular structures and vigorous phagocytosis of those organisms.

  20. Protein phosphatase 2B (PP2B, calcineurin) in Paramecium: partial characterization reveals that two members of the unusually large catalytic subunit family have distinct roles in calcium-dependent processes.

    PubMed

    Fraga, D; Sehring, I M; Kissmehl, R; Reiss, M; Gaines, R; Hinrichsen, R; Plattner, H

    2010-07-01

    We characterized the calcineurin (CaN) gene family, including the subunits CaNA and CaNB, based upon sequence information obtained from the Paramecium genome project. Paramecium tetraurelia has seven subfamilies of the catalytic CaNA subunit and one subfamily of the regulatory CaNB subunit, with each subfamily having two members of considerable identity on the amino acid level (>or=55% between subfamilies, >or=94% within CaNA subfamilies, and full identity in the CaNB subfamily). Within CaNA subfamily members, the catalytic domain and the CaNB binding region are highly conserved and molecular modeling revealed a three-dimensional structure almost identical to a human ortholog. At 14 members, the size of the CaNA family is unprecedented, and we hypothesized that the different CaNA subfamily members were not strictly redundant and that at least some fulfill different roles in the cell. This was tested by selecting two phylogenetically distinct members of this large family for posttranscriptional silencing by RNA interference. The two targets resulted in differing effects in exocytosis, calcium dynamics, and backward swimming behavior that supported our hypothesis that the large, highly conserved CaNA family members are not strictly redundant and that at least two members have evolved diverse but overlapping functions. In sum, the occurrence of CaN in Paramecium spp., although disputed in the past, has been established on a molecular level. Its role in exocytosis and ciliary beat regulation in a protozoan, as well as in more complex organisms, suggests that these roles for CaN were acquired early in the evolution of this protein family.

  1. Protein Phosphatase 2B (PP2B, Calcineurin) in Paramecium: Partial Characterization Reveals That Two Members of the Unusually Large Catalytic Subunit Family Have Distinct Roles in Calcium-Dependent Processes▿‡

    PubMed Central

    Fraga, D.; Sehring, I. M.; Kissmehl, R.; Reiss, M.; Gaines, R.; Hinrichsen, R.; Plattner, H.

    2010-01-01

    We characterized the calcineurin (CaN) gene family, including the subunits CaNA and CaNB, based upon sequence information obtained from the Paramecium genome project. Paramecium tetraurelia has seven subfamilies of the catalytic CaNA subunit and one subfamily of the regulatory CaNB subunit, with each subfamily having two members of considerable identity on the amino acid level (≥55% between subfamilies, ≥94% within CaNA subfamilies, and full identity in the CaNB subfamily). Within CaNA subfamily members, the catalytic domain and the CaNB binding region are highly conserved and molecular modeling revealed a three-dimensional structure almost identical to a human ortholog. At 14 members, the size of the CaNA family is unprecedented, and we hypothesized that the different CaNA subfamily members were not strictly redundant and that at least some fulfill different roles in the cell. This was tested by selecting two phylogenetically distinct members of this large family for posttranscriptional silencing by RNA interference. The two targets resulted in differing effects in exocytosis, calcium dynamics, and backward swimming behavior that supported our hypothesis that the large, highly conserved CaNA family members are not strictly redundant and that at least two members have evolved diverse but overlapping functions. In sum, the occurrence of CaN in Paramecium spp., although disputed in the past, has been established on a molecular level. Its role in exocytosis and ciliary beat regulation in a protozoan, as well as in more complex organisms, suggests that these roles for CaN were acquired early in the evolution of this protein family. PMID:20435698

  2. Evolutionary rescue and local adaptation under different rates of temperature increase: a combined analysis of changes in phenotype expression and genotype frequency in Paramecium microcosms.

    PubMed

    Killeen, Joshua; Gougat-Barbera, Claire; Krenek, Sascha; Kaltz, Oliver

    2017-04-01

    Evolutionary rescue (ER) occurs when populations, which have declined due to rapid environmental change, recover through genetic adaptation. The success of this process and the evolutionary trajectory of the population strongly depend on the rate of environmental change. Here we investigated how different rates of temperature increase (from 23 to 32 °C) affect population persistence and evolutionary change in experimental microcosms of the protozoan Paramecium caudatum. Consistent with theory on ER, we found that those populations experiencing the slowest rate of temperature increase were the least likely to become extinct and tended to be the best adapted to the new temperature environment. All high-temperature populations were more tolerant to severe heat stress (35, 37 °C), indicating a common mechanism of heat protection. High-temperature populations also had superior growth rates at optimum temperatures, leading to the absence of a pattern of local adaptation to control (23 °C) and high-temperature (32 °C) environments. However, high-temperature populations had reduced growth at low temperatures (5-9 °C), causing a shift in the temperature niche. In part, the observed evolutionary change can be explained by selection from standing variation. Using mitochondrial markers, we found complete divergence between control and high-temperature populations in the frequencies of six initial founder genotypes. Our results confirm basic predictions of ER and illustrate how adaptation to an extreme local environment can produce positive as well as negative correlated responses to selection over the entire range of the ecological niche. © 2017 John Wiley & Sons Ltd.

  3. Synthesis of the signal molecule acetylcholine during the developmental cycle of Paramecium primaurelia (Protista, Ciliophora) and its possible function in conjugation.

    PubMed

    Delmonte Corrado, M U; Politi, H; Ognibene, M; Angelini, C; Trielli, F; Ballarini, P; Falugi, C

    2001-06-01

    We recently discovered, in mating-competent Paramecium primaurelia, the presence of functionally related molecules of the cholinergic system: the neurotransmitter acetylcholine (ACh), both its nicotinic and muscarinic receptors and its lytic enzyme acetylcholinesterase (AChE). Our results on the inhibition of mating-cell pairing in vivo in mating-competent cells treated with cholinomimetic drugs support the hypothesis that the cholinergic system plays a role in cell-to-cell adhesion. To investigate the possible function of the signal molecule ACh in conjugation in P. primaurelia, we attempted to detect the intracellular sites of ACh synthesis by localizing the ACh biosynthetic enzyme choline acetyltransferase (ChAT). Using immunocytochemical and histochemical methods, we have demonstrated the presence and activity of ChAT principally on the surface membrane of mating-competent cells and of mature but non-mating-competent cells. No evidence for ChAT activity was found in immature cells. Immunoblot analysis revealed the presence of immunoreactive bands, ranging in molecular mass from 42 to 133 kDa, as reported for ChAT isolated from higher organisms. In vivo experiments showed that inhibition of ChAT activity by Congo Red, known to be a potent competitive inhibitor of acetyl coenzyme A, did not affect mating-cell pairing. Conversely, inhibition of AChE with BW 284c51 or eserine, which block enzyme activity by reacting with a specific serine within the catalytic centre, significantly inhibited mating-cell pairing. Our results suggest that ACh has a negative modulating effect on conjugation in P. primaurelia.

  4. Application of fluorescence resonance energy transfer techniques to the study of lectin-binding site distribution on Paramecium primaurelia (Protista, Ciliophora) cell surface.

    PubMed

    Locatelli, D; Delmonte Corrado, M U; Politi, H; Bottiroli, G

    1998-01-01

    Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon occurring between the molecules of two fluorochromes with suitable spectral characteristics (donor-acceptor dye pair), and consisting in an excitation energy migration through a non-radiative process. Since the efficiency of the process is strictly dependent on the distance and reciprocal orientation of the donor and acceptor molecules, FRET-based techniques can be successfully applied to the study of biomolecules and cell component organisation and distribution. These techniques have been employed in studying Paramecium primaurelia surface membrane for the reciprocal distribution of N-acetylneuraminic acid (NeuAc) and N-acetylglucosamine (GlcNAc) glycosidic residues, which were found to be involved in mating cell pairing. NeuAc and GlcNAc were detected by their specific binding lectins, Limulus polyphemus agglutinin (LPA) and wheat germ agglutinin (WGA), respectively. Microspectrofluorometric analysis afforded the choice of fluorescein isothiocyanate and Texas red conjugated with LPA and WGA, respectively, as a suitable donor-acceptor couple efficiently activating FRET processes. Studies performed both in solution and in cells allowed to define the experimental conditions favourable for a FRET analysis. The comparative study carried out both on the conjugating-region and the non conjugating region of the surface membrane, indicates that FRET distribution appears quite homogeneous in mating-competent mating type (mt) I, whereas, in mating-competent mt II cells, FRET distribution seems to be preferentially localised on the conjugating-region functionally involved in mating cell pairing. This difference in the distribution of lectin-binding sites is suggested to be related to mating-competence acquisition.

  5. Harm reduction

    PubMed Central

    Normand, Jacques; Li, Jih-Heng; Thomson, Nicholas; Jarlais, Don Des

    2014-01-01

    The “Harm Reduction” session was chaired by Dr. Jacques Normand, Director of the AIDS Research Program of the U.S. National Institute on Drug Abuse. The three presenters (and their presentation topics) were: Dr. Don Des Jarlais (High Coverage Needle/Syringe Programs for People Who Inject Drugs in Low and Middle Income Countries: A Systematic Review), Dr. Nicholas Thomson (Harm Reduction History, Response, and Current Trends in Asia), and Dr. Jih-Heng Li (Harm Reduction Strategies in Taiwan). PMID:25278732

  6. KIN241: a gene involved in cell morphogenesis in Paramecium tetraurelia reveals a novel protein family of cyclophilin-RNA interacting proteins (CRIPs) conserved from fission yeast to man.

    PubMed

    Krzywicka, A; Beisson, J; Keller, A M; Cohen, J; Jerka-Dziadosz, M; Klotz, C

    2001-10-01

    In this study, we report cloning, by functional complementation of the KIN241 gene involved in Paramecium cell morphogenesis, cortical organization and nuclear reorganization. This gene is predicted to encode a protein of a novel type, comprising a cyclophilin-type, peptidyl-prolyl isomerase domain, an RNA recognition motif, followed by a region rich in glutamate and lysine (EK domain) and a C-terminal string of serines. As homologues of this protein are present in the genomes of Schizosaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Arabidopsis thaliana and Homo sapiens, the Kin241p predicted sequence defines a new family of proteins that we propose to call 'CRIP', for cyclophilin-RNA interacting protein. We demonstrate that, in Paramecium, Kin241p is localized in the nucleus and that deletion of some nuclear localization signals (NLSs) decreases transport of the protein into the nucleus. No Kin241-1 protein is present in mutant cells, suggesting that the C-terminal serine-rich region is responsible for protein stability.

  7. Waste Reduction.

    ERIC Educational Resources Information Center

    Bray, Marilyn; And Others

    1996-01-01

    Presents activities that focus on waste reduction in the school and community. The ideas are divided into grade level categories. Sample activities include Techno-Trash, where children use tools to take apart broken appliances or car parts, then reassemble them or build new creations. Activities are suggested for areas including language arts and…

  8. Reduction Corporoplasty

    PubMed Central

    Hakky, Tariq S.; Martinez, Daniel; Yang, Christopher; Carrion, Rafael E.

    2015-01-01

    Objective Here we present the first video demonstration of reduction corporoplasty in the management of phallic disfigurement in a 17 year old man with a history sickle cell disease and priapism. Introduction Surgical management of aneurysmal dilation of the corpora has yet to be defined in the literature. Materials and Methods: We preformed bilateral elliptical incisions over the lateral corpora as management of aneurysmal dilation of the corpora to correct phallic disfigurement. Results The patient tolerated the procedure well and has resolution of his corporal disfigurement. Conclusions Reduction corporoplasty using bilateral lateral elliptical incisions in the management of aneurysmal dilation of the corpora is a safe an feasible operation in the management of phallic disfigurement. PMID:26005988

  9. Nitrate reduction

    DOEpatents

    Dziewinski, Jacek J.; Marczak, Stanislaw

    2000-01-01

    Nitrates are reduced to nitrogen gas by contacting the nitrates with a metal to reduce the nitrates to nitrites which are then contacted with an amide to produce nitrogen and carbon dioxide or acid anions which can be released to the atmosphere. Minor amounts of metal catalysts can be useful in the reduction of the nitrates to nitrites. Metal salts which are formed can be treated electrochemically to recover the metals.

  10. Simulation model of Cryptomonas ovata population dynamics in southern Kootenay Lake, British Columbia

    USGS Publications Warehouse

    Cloern, James E.

    1978-01-01

    The model simulates well the timing and magnitude of all observed population changes and, more importantly, it gives insight into the important mechanisms which regulate population density of C. ovata in this natural system.

  11. Effects of diuron and carbofuran pesticides in their pure and commercial forms on Paramecium caudatum: The use of protozoan in ecotoxicology.

    PubMed

    Mansano, Adrislaine S; Moreira, Raquel A; Pierozzi, Mayara; Oliveira, Thiessa M A; Vieira, Eny M; Rocha, Odete; Regali-Seleghim, Mirna H

    2016-06-01

    Toxic effects of diuron and carbofuran on Paramecium caudatum were evaluated. Acute and chronic tests were conducted with diuron and carbofuran active ingredients and their commercial formulations, Diuron Nortox(®) 500 SC and Furadan(®) 350 SC, respectively. The sensitivity range of P. caudatum to reference substance sodium chloride was established. A preliminary risk assessment of diuron and carbofuran for Brazilian water bodies was performed. The tests indicated that toxicity of pure diuron and its commercial formulation was similar, while the commercial product carbofuran was more toxic than its pure form. In acute tests, readings were carried out at 2, 3, 4 and 6 h and showed an increase of mortality with increasing exposure time. The sensitivity of P. caudatum to NaCl ranged from 3.31 to 4.44 g L(-1), averaging 3.88 g L(-1). For diuron, the 6 h LC50 was 64.6 ± 3.3 mg L(-1) for its pure form and 62.4 ± 2.5 mg L(-1) for its commercial formulation. Carbofuran active ingredient was less toxic than that of diuron, presenting a 6 h LC50 of 142.0 ± 2.4 mg L(-1) for its pure form and 70.4 ± 2.2 mg L(-1) for its commercial product. Chronic tests showed that these pesticides cause significant decrease on population growth, generation number and biomass of P. caudatum. The 24 h IC50 was 7.10 ± 0.58 mg L(-1) for pure diuron, 6.78 ± 0.92 mg L(-1) for commercial diuron, 22.95 ± 3.57 mg L(-1) for pure carbofuran and 4.98 ± 0.62 mg L(-1) for commercial carbofuran. Preliminary risk assessment indicated that diuron and carbofuran present potential ecological risks for Brazilian water bodies. P. caudatum was a suitable and sensitive test organism to evaluate diuron and carbofuran toxicity to freshwater protozooplankton and, taking into account the relevant role of protozoans in aquatic environments, we strongly recommend its inclusion in ecotoxicological studies.

  12. A key function of non-planar membranes and their associated microtubular ribbons in contractile vacuole membrane dynamics is revealed by electrophysiologically controlled fixation of Paramecium.

    PubMed

    Tominaga, T; Naitoh, Y; Allen, R D

    1999-11-01

    The contractile vacuole complex of the fresh water protozoan Paramecium multimicronucleatum exhibits periodic exocytotic activity. This keeps cytosolic osmolarity at a constant value. The contractile vacuole, the central exocytotic vesicle of the complex, becomes disconnected from its surrounding radial arms and rounds before its fluid content is expelled. We previously proposed a hypothesis that the rounding of the contractile vacuole corresponds to an increase in its membrane tension and that a periodic increase in membrane tension governs the exocytotic cycle. We also proposed a hypothesis that transformation of excess planar membrane of the contractile vacuole into 40 nm diameter tubules, that remain continuous with the contractile vacuole membrane, is a primary cause for the tension development in the planar membrane. In order to investigate tension development further, we have examined electron microscopically the contractile vacuole membrane at the rounding phase. To do this, we developed a computer-aided system to fix the cell precisely at the time that the contractile vacuole exhibited rounding. In this system a decrease in the electrical potential across the contractile vacuole membrane that accompanied the vacuole's rounding was monitored through a fine-tipped microelectrode inserted directly into the in vivo contractile vacuole. A decrease in membrane potential was used to generate an electric signal that activated an injector for injecting a fixative through a microcapillary against the cell at the precise time of rounding. Subsequent electron micrographs of the contractile vacuole membrane clearly demonstrated that numerous approximately 40 nm membrane-bound tubules formed in the vicinity of the vacuole's microtubule ribbons when the vacuole showed rounding. This finding suggested that membrane tubulation was the cause for topographical isolation of excess membrane from the planar membrane during the periodic rounding of the contractile vacuole. This

  13. Two distinct classes of mitotic cyclin homologues, Cyc1 and Cyc2, are involved in cell cycle regulation in the ciliate Paramecium tetraurelia.

    PubMed

    Zhang, H; Adl, S M; Berger, J D

    1999-01-01

    The eukaryotic cell cycle is regulated by the sequential activation of different CDK/cyclin complexes. Two distinct classes of mitotic cyclin homologues, CYC1 and CYC2, have been identified and cloned for the first time in the ciliate Paramecium. Cyc1 is 324 amino acids long with a predicted molecular mass of 38 kDa, whereas Cyc2 is 336 amino acids long with a predicted molecular mass of 40 kDa. They display 42-51% sequence identity to other eukaryotic mitotic cyclins within the 'cyclin box' region. The conserved 'cyclin box' and 'destruction box' elements can be identified within each of the sequences. Genomic Southern blot analysis indicated that the CYC1 gene has two isoforms, with 92.3% and 85.9% identify at the amino acid level and at the nucleotide level, respectively. Both Cyc1 and Cyc2 proteins showed characteristic patterns of accumulation and destruction during the vegetative cell cycle, with Cyc1 peaking at the point of commitment to division (PCD), and Cyc2 reaching the maximal level late in the cell cycle. Immunoprecipitation experiments with antibodies specific to Cyc1 and Cyc2 indicated that Cyc1 and Cyc2 associate with distinct CDK homologues. Both immunoprecipitates exhibited histone H1 kinase activity that oscillated in the cell cycle in parallel with the respective amount of cyclins present. Histone H1 kinase activity associated with Cyc1 reached a peak at PCD while Cyc2 showed maximal activity when about 75% cells have completed cytokinesis. We propose that Cyc1 may be involved in commitment to division, in association with the CDK that binds to p13suc1, Cdk3, and that the Cyc2/Cdk2 complex may regulate cytokinesis. PCR-amplification revealed similar sequences in Tetrahymena, Sterkiella, Colpoda and Blepharisma, suggesting the conservation of the cyclin genes within ciliates. Although cell cycle regulation in ciliates differs in some respects from that of other eukaryotes, the cyclin motifs have clearly been conserved during evolution.

  14. The role of the micronucleus in stomatogenesis in sexual reproduction of Paramecium tetraurelia: laser microbeam irradiation of the micronucleus.

    PubMed

    Tam, L W; Ng, S F

    1986-12-01

    Fifteen amicronucleate cell lines and 22 cell lines with defective micronuclei were obtained following selective laser microbeam irradiation of the micronucleus. The amicronucleate cell lines showed reduced growth rate and formed abnormal oral apparatuses in asexual reproduction, and failed to produce any oral apparatus in autogamy, in agreement with previous observations on amicronucleate cells obtained by micropipetting. The 22 cell lines with defective micronucleus exhibited various abnormalities of the oral apparatus newly formed during autogamy. These abnormalities included the arrest of membranelle assembly, reduction in the length of the buccal cavity and oral membranelles, disruption of the organization of the membranelles, quadrulation of the dorsal peniculus, and failure of addition of membranellar basal body rows. Hence the micronucleus plays multiple roles in sexual stomatogenesis, deciding early steps of oral membranelle assembly and affecting their subsequent patterning. Our results agree with the notion that the micronucleus acts during a critical period between the second meiotic division and up to the formation of the zygotic nucleus to control the early stage of oral membranelle assembly. Laser microbeam irradiation might have created recessive mutations and/or chromosomal aberrations, which were expressed during this critical period with the formation of abnormal postmeiotic nuclei.

  15. Physiological consequences of mitochondrial antibiotic-resistant mutations in Paramecium: growth-rates, cytochromic defects and cyanide-insensitive respiration of mutant and erythromycin-treated wild-type strains.

    PubMed

    Adoutte, A; Doussiere, J

    1978-05-03

    A set of mitochondrial antibiotic-resistant mutants of Paramecium have been analyzed with respect to their growth-rates, cytochromic content and respiratory properties. The mutants could be arranged in a continuous series ranging from strains equivalent to wild-type to severely affected ones; affected strains display longer generation times, reduced amount of cytochrome oxidase and very high levels of cyanideinsensitive respiration. Perfect phenocopies of the mutants were obtained by treating wild-type cells with low concentrations of erythromycin suggesting that the mutations exert their pleiotropic effect by perturbating mitochondria protein synthesis in agreement with the idea that these mutations affect the mitochondrial ribosomes. In the mitochondria of some of the mutants, electrons can be channelled with equal efficiency into the "classical" cyanide-sensitive pathway and the alternate cyanide insensitive (and SHAM-sensitive) one, providing direct demonstration of the branching of these two respiratory pathways. In the absence of any added inhibitor, however, electrons tend to be channelled in the cyanide-sensitive pathway. All the physiological data fit perfectly the genetic data concerning the "stability" of the various mutations in "mixed mitochondrial populations", i.e., markers that were known to be strongly counter-selected with respect to wild-type in such populations correspond to severely affected strains, while markers that were known to be "stable" correspond to "healthy" strains. A more quantitative analysis of the data shows that that there is little or no "complementation" between wild-type and mutated mitochondria in mixed cells indicating a high extent of functional autonomy of mitochondria in Paramecium.

  16. Site-Directed Mutagenesis, in Vivo Electroporation and Mass Spectrometry in Search for Determinants of the Subcellular Targeting of Rab7b Paralogue in the Model Eukaryote Paramecium Octaurelia

    PubMed Central

    Wyroba, E.; Kwaśniak, P.; Miller, K.; Kobyłecki, K.; Osińska, M.

    2016-01-01

    Protein products of paralogous genes resulting from whole genome duplication may acquire new functions. The role of post-translational modifications (PTM) in proper targeting of Paramecium Rab7b paralogue (distinct from that of Rab7a directly involved in phagocytosis) was studied using point mutagenesis, proteomic analysis and double immunofluorescence after in vivo electroporation of the mutagenized protein. Here we show that substitution of Thr200 by Ala diminished the incorporation of [P32] by 37% and of [C14-]UDP-glucose by 24% into recombinant Rab7b_200 in comparison to the non-mutagenized control. Double confocal imaging revealed that Rab7b_200 was mistargeted upon electroporation into living cells in contrast to non-mutagenized recombinant Rab7b correctly incorporated in the cytostome area. Using nano LC-MS/MS to compare the peptide map of Rab7b with that after deglycosylation with a mixture of five enzymes of different specificity we identified a peptide ion at m/z=677.63+ representing a glycan group attached to Thr200. Based on its mass and quantitative assays with [P32] and [C14]UDP-glucose, the suggested composition of the adduct attached to Thr200 is (Hex)1(HexNAc)1(Phos)3 or (HexNAc)1 (Deoxyhexose)1 (Phos)1 (HexA)1. These data indicate that PTM of Thr200 located in the hypervariable C-region of Paramecium octaurelia Rab7b is crucial for the proper localization/function of this protein. Moreover, the two Rab7 paralogues differ also in another PTM: substantially more phosphorylated amino acid residues are in Rab7b than in Rab7a. PMID:27349314

  17. Site-directed mutagenesis, in vivo electroporation and mass spectrometry in search for determinants of the subcellular targeting of Rab7b paralogue in the model eukaryote Paramecium octaurelia.

    PubMed

    Wyroba, E; Kwaśniak, P; Miller, K; Kobyłecki, K; Osińska, M

    2016-04-11

    Protein products of the paralogous genes resulting from the whole genome duplication may acquire new function. The role of post-translational modifications (PTM) in proper targeting of Paramecium Rab7b paralogue - distinct from that of Rab7a directly involved in phagocytosis - was studied using point mutagenesis, proteomic analysis and double immunofluorescence after in vivo electroporation of the mutagenized protein. Here we show that substitution of Thr200 by Ala200 resulted in diminished incorporation of [P32] by 37.4% and of 32 [C14-]UDP-glucose by 24%, respectively, into recombinant Rab7b_200 in comparison to the non-mutagenized control. Double confocal imaging revealed that Rab7b_200 was mistargeted upon electroporation into living cells contrary to non- mutagenized recombinant Rab7b correctly incorporated in the cytostome area. We identified the peptide ion at m/z=677.63+ characteristic for the glycan group attached to Thr200 in Rab7b using nano LC-MS/MS and comparing the peptide map of this protein with that after deglycosylation with the mixture of five enzymes of different specificity. Based on the mass of this peptide ion and quantitative radioactive assays with [P32]and  [C14-]UDP- glucose, the suggested composition of the adduct attached to Thr200 might be (Hex)1(HexNAc)1(Phos)3 or (HexNAc)1 (Deoxyhexose)1 (Phos)1 (HexA)1. These data indicate that PTM of Thr200 located in the hypervariable C-region of Rab7b in Paramecium is crucial for the proper localization/function of this protein. Moreover, these proteins differ also in other PTM: the number of phosphorylated amino acids in Rab7b is much higher than in Rab7a.

  18. Disentangling the Taxonomy of Rickettsiales and Description of Two Novel Symbionts (“Candidatus Bealeia paramacronuclearis” and “Candidatus Fokinia cryptica”) Sharing the Cytoplasm of the Ciliate Protist Paramecium biaurelia

    PubMed Central

    Szokoli, Franziska; Castelli, Michele; Sabaneyeva, Elena; Schrallhammer, Martina; Krenek, Sascha; Doak, Thomas G.; Berendonk, Thomas U.

    2016-01-01

    ABSTRACT In the past 10 years, the number of endosymbionts described within the bacterial order Rickettsiales has constantly grown. Since 2006, 18 novel Rickettsiales genera inhabiting protists, such as ciliates and amoebae, have been described. In this work, we characterize two novel bacterial endosymbionts from Paramecium collected near Bloomington, IN. Both endosymbiotic species inhabit the cytoplasm of the same host. The Gram-negative bacterium “Candidatus Bealeia paramacronuclearis” occurs in clumps and is frequently associated with the host macronucleus. With its electron-dense cytoplasm and a distinct halo surrounding the cell, it is easily distinguishable from the second smaller symbiont, “Candidatus Fokinia cryptica,” whose cytoplasm is electron lucid, lacks a halo, and is always surrounded by a symbiontophorous vacuole. For molecular characterization, the small-subunit rRNA genes were sequenced and used for taxonomic assignment as well as the design of species-specific oligonucleotide probes. Phylogenetic analyses revealed that “Candidatus Bealeia paramacronuclearis” clusters with the so-called “basal” Rickettsiales, and “Candidatus Fokinia cryptica” belongs to “Candidatus Midichloriaceae.” We obtained tree topologies showing a separation of Rickettsiales into at least two groups: one represented by the families Rickettsiaceae, Anaplasmataceae, and “Candidatus Midichloriaceae” (RAM clade), and the other represented by “basal Rickettsiales,” including “Candidatus Bealeia paramacronuclearis.” Therefore, and in accordance with recent publications, we propose to limit the order Rickettsiales to the RAM clade and to raise “basal Rickettsiales” to an independent order, Holosporales ord. nov., inside Alphaproteobacteria, which presently includes four family-level clades. Additionally, we define the family “Candidatus Hepatincolaceae” and redefine the family Holosporaceae. IMPORTANCE In this paper, we provide the

  19. Breast Reduction Surgery

    MedlinePlus

    ... considering breast reduction surgery, consult a board-certified plastic surgeon. It's important to understand what breast reduction ... risk of complications from breast reduction surgery. Your plastic surgeon will likely: Evaluate your medical history and ...

  20. Symbiotic Chlorella variabilis incubated under constant dark conditions for 24 hours loses the ability to avoid digestion by host lysosomal enzymes in digestive vacuoles of host ciliate Paramecium bursaria.

    PubMed

    Kodama, Yuuki; Fujishima, Masahiro

    2014-12-01

    Endosymbiosis between symbiotic Chlorella and alga-free Paramecium bursaria cells can be induced by mixing them. To establish the endosymbiosis, algae must acquire temporary resistance to the host lysosomal enzymes in the digestive vacuoles (DVs). When symbiotic algae isolated from the alga-bearing paramecia are kept under a constant dark conditions for 24 h before mixing with the alga-free paramecia, almost all algae are digested in the host DVs. To examine the cause of algal acquisition to the host lysosomal enzymes, the isolated algae were kept under a constant light conditions with or without a photosynthesis inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea for 24 h, and were mixed with alga-free paramecia. Unexpectedly, most of the algae were not digested in the DVs irrespective of the presence of the inhibitor. Addition of 1 mM maltose, a main photosynthetic product of the symbiotic algae or of a supernatant of the isolated algae kept for 24 h under a constant light conditions, did not rescue the algal digestion in the DVs. These observations reveal that unknown factors induced by light are a prerequisite for algal resistance to the host lysosomal enzymes. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  1. Differences in infectivity between endosymbiotic Chlorella variabilis cultivated outside host Paramecium bursaria for 50 years and those immediately isolated from host cells after one year of reendosymbiosis

    PubMed Central

    Kodama, Y.; Fujishima, M.

    2016-01-01

    ABSTRACT Chlorella variabilis strain NC64A is an intracellular photobiont of the ciliate Paramecium bursaria. NC64A was isolated from P. bursaria nearly 50 years ago and was thereafter cultivated outside the host. This study was undertaken to detect changes in its infectivity to P. bursaria and its auxotrophy for growth outside the host induced during long-term cultivation. NC64A can grow in Modified Bold's Basal Medium but not in C medium, whereas another symbiotic Chlorella variabilis strain, 1N, that was recently isolated from the host grew in C medium but not in Modified Bold's Basal Medium. With regards infectivity, NC64A in the logarithmic phase of growth showed low infectivity to alga-removed P. bursaria cells, whereas those in the early stationary phase showed high infectivity of about 30%. Those in the decay phase of growth showed no infectivity. Results show that NC64A has infectivity, but the infection rate depends on their culture age in the growth curve. Furthermore, NC64A that had been re-infected to P. bursaria for more than one year and isolated from the host showed a nearly 100% infection rate, which indicates that NC64A can recover its infectivity by re-infection to P. bursaria. PMID:26718931

  2. Reductive dissolution of goethite by phenolic reductants

    SciTech Connect

    LaKind, J.S.; Stone, A.T. )

    1989-05-01

    The reductive dissolution of goethite ({alpha}-FeOOH) and hematite ({alpha}-Fe{sub 2}O{sub 3}) by phenolic reductants has been examined in order to improve the understanding of iron transformations in soils, sediments and aquifers. Rates of goethite reductive dissolution by hydroquinone increased as the pH was increased from pH 1.8 to 4.65, and the following reaction stoichiometry was obeyed: 2{alpha}-FeOOH + QH{sub 2} = 2Fe{sup 2+} + Q + 4OH{sup {minus}}. As the pH was increased from pH 4.5 to 6.0, the reductive dissolution rate decreased to below the detection limit. At pH 3.4, the reductive dissolution of hematite was two orders of magnitude slower than goethite. The relationship between structure and reactivity was examined for a series of mono-, di-, and tri-hydroxybenzene reductants. Rates of reductive dissolution decreased in the following order: catechol {approx equal} hydroquinone > 3,4-dihydroxybenzoic acid > resorcinol-phenol-4-hydroxybenzoic acid.

  3. Local reduction in physics

    NASA Astrophysics Data System (ADS)

    Rosaler, Joshua

    2015-05-01

    A conventional wisdom about the progress of physics holds that successive theories wholly encompass the domains of their predecessors through a process that is often called "reduction." While certain influential accounts of inter-theory reduction in physics take reduction to require a single "global" derivation of one theory's laws from those of another, I show that global reductions are not available in all cases where the conventional wisdom requires reduction to hold. However, I argue that a weaker "local" form of reduction, which defines reduction between theories in terms of a more fundamental notion of reduction between models of a single fixed system, is available in such cases and moreover suffices to uphold the conventional wisdom. To illustrate the sort of fixed-system, inter-model reduction that grounds inter-theoretic reduction on this picture, I specialize to a particular class of cases in which both models are dynamical systems. I show that reduction in these cases is underwritten by a mathematical relationship that follows a certain liberalized construal of Nagel/Schaffner reduction, and support this claim with several examples. Moreover, I show that this broadly Nagelian analysis of inter-model reduction encompasses several cases that are sometimes cited as instances of the "physicist's" limit-based notion of reduction.

  4. Reduction Mechanisms in Manganese Ore Reduction

    NASA Astrophysics Data System (ADS)

    Coetsee, Theresa; Reinke, Christian; Nell, Johannes; Pistorius, Petrus Christiaan

    2015-12-01

    Manganese ores are highly heterogeneous and contain various minerals with different levels of contained manganese and iron and therefore the ore reduction behavior is not uniform. Both phase chemistry and phase morphology at the reaction interface, at micron scale, must be investigated to understand the reaction mechanism effects in manganese ore reduction. This approach is applied here to reacted material mixture samples taken from the AlloyStream pilot plant furnace over a period of 4 months. The mineralogical features are reported and discussed. Deductions are made on the likely dominant reduction mechanism in this reaction system, given the phase morphology observations presented.

  5. Breast reduction (mammoplasty) - slideshow

    MedlinePlus

    ... page: //medlineplus.gov/ency/presentations/100189.htm Breast reduction (mammoplasty) - series—Indications To use the sharing features ... to slide 4 out of 4 Overview Breast reduction is usually performed for enlarged breasts (macromastia), but ...

  6. Solving Problems Reductively

    ERIC Educational Resources Information Center

    Armoni, Michal; Gal-Ezer, Judith; Tirosh, Dina

    2005-01-01

    Solving problems by reduction is an important issue in mathematics and science education in general (both in high school and in college or university) and particularly in computer science education. Developing reductive thinking patterns is an important goal in any scientific discipline, yet reduction is not an easy subject to cope with. Still,…

  7. Drag reduction in nature

    NASA Technical Reports Server (NTRS)

    Bushnell, D. M.; Moore, K. J.

    1991-01-01

    Recent studies on the drag-reducing shapes, structures, and behaviors of swimming and flying animals are reviewed, with an emphasis on potential analogs in vehicle design. Consideration is given to form drag reduction (turbulent flow, vortex generation, mass transfer, and adaptations for body-intersection regions), skin-friction drag reduction (polymers, surfactants, and bubbles as surface 'additives'), reduction of the drag due to lift, drag-reduction studies on porpoises, and drag-reducing animal behavior (e.g., leaping out of the water by porpoises). The need for further research is stressed.

  8. Drag reduction in nature

    NASA Technical Reports Server (NTRS)

    Bushnell, D. M.; Moore, K. J.

    1991-01-01

    Recent studies on the drag-reducing shapes, structures, and behaviors of swimming and flying animals are reviewed, with an emphasis on potential analogs in vehicle design. Consideration is given to form drag reduction (turbulent flow, vortex generation, mass transfer, and adaptations for body-intersection regions), skin-friction drag reduction (polymers, surfactants, and bubbles as surface 'additives'), reduction of the drag due to lift, drag-reduction studies on porpoises, and drag-reducing animal behavior (e.g., leaping out of the water by porpoises). The need for further research is stressed.

  9. CARBON DIOXIDE REDUCTION SYSTEM.

    DTIC Science & Technology

    CARBON DIOXIDE , *SPACE FLIGHT, RESPIRATION, REDUCTION(CHEMISTRY), RESPIRATION, AEROSPACE MEDICINE, ELECTROLYSIS, INSTRUMENTATION, ELECTROLYTES, VOLTAGE, MANNED, YTTRIUM COMPOUNDS, ZIRCONIUM COMPOUNDS, NICKEL.

  10. Managing Faculty Reductions

    ERIC Educational Resources Information Center

    Alm, Kent F.; And Others

    1977-01-01

    A process for the management of reductions in the number of faculty positions available to a university is described. It considers staffing by projections, the evolution of personnel planning, and the balance of reductions in faculty and administration, along with coping strategies and advice growing out of five years of enrollment decline…

  11. Microbial reductive dehalogenation.

    PubMed Central

    Mohn, W W; Tiedje, J M

    1992-01-01

    A wide variety of compounds can be biodegraded via reductive removal of halogen substituents. This process can degrade toxic pollutants, some of which are not known to be biodegraded by any other means. Reductive dehalogenation of aromatic compounds has been found primarily in undefined, syntrophic anaerobic communities. We discuss ecological and physiological principles which appear to be important in these communities and evaluate how widely applicable these principles are. Anaerobic communities that catalyze reductive dehalogenation appear to differ in many respects. A large number of pure cultures which catalyze reductive dehalogenation of aliphatic compounds are known, in contrast to only a few organisms which catalyze reductive dehalogenation of aromatic compounds. Desulfomonile tiedjei DCB-1 is an anaerobe which dehalogenates aromatic compounds and is physiologically and morphologically unusual in a number of respects, including the ability to exploit reductive dehalogenation for energy metabolism. When possible, we use D. tiedjei as a model to understand dehalogenating organisms in the above-mentioned undefined systems. Aerobes use reductive dehalogenation for substrates which are resistant to known mechanisms of oxidative attack. Reductive dehalogenation, especially of aliphatic compounds, has recently been found in cell-free systems. These systems give us an insight into how and why microorganisms catalyze this activity. In some cases transition metal complexes serve as catalysts, whereas in other cases, particularly with aromatic substrates, the catalysts appear to be enzymes. Images PMID:1406492

  12. Does Source Reduction Work?

    ERIC Educational Resources Information Center

    Allaway, David

    1992-01-01

    Suggests that quantification is essential to establish the cost-effectiveness of source reduction (SR). Presents case studies of monitoring methods for seven different kinds of SR efforts: (1) packaging changes, (2) SR businesses, (3) waste exchanges, (4) individual nonresidential efforts, (5) variable garbage rates, (6) yard waste reduction, and…

  13. Does Source Reduction Work?

    ERIC Educational Resources Information Center

    Allaway, David

    1992-01-01

    Suggests that quantification is essential to establish the cost-effectiveness of source reduction (SR). Presents case studies of monitoring methods for seven different kinds of SR efforts: (1) packaging changes, (2) SR businesses, (3) waste exchanges, (4) individual nonresidential efforts, (5) variable garbage rates, (6) yard waste reduction, and…

  14. Bayesian supervised dimensionality reduction.

    PubMed

    Gönen, Mehmet

    2013-12-01

    Dimensionality reduction is commonly used as a preprocessing step before training a supervised learner. However, coupled training of dimensionality reduction and supervised learning steps may improve the prediction performance. In this paper, we introduce a simple and novel Bayesian supervised dimensionality reduction method that combines linear dimensionality reduction and linear supervised learning in a principled way. We present both Gibbs sampling and variational approximation approaches to learn the proposed probabilistic model for multiclass classification. We also extend our formulation toward model selection using automatic relevance determination in order to find the intrinsic dimensionality. Classification experiments on three benchmark data sets show that the new model significantly outperforms seven baseline linear dimensionality reduction algorithms on very low dimensions in terms of generalization performance on test data. The proposed model also obtains the best results on an image recognition task in terms of classification and retrieval performances.

  15. Microbial reduction of uranium

    USGS Publications Warehouse

    Lovley, D.R.; Phillips, E.J.P.; Gorby, Y.A.; Landa, E.R.

    1991-01-01

    REDUCTION of the soluble, oxidized form of uranium, U(VI), to insoluble U(IV) is an important mechanism for the immobilization of uranium in aquatic sediments and for the formation of some uranium ores1-10. U(VI) reduction has generally been regarded as an abiological reaction in which sulphide, molecular hydrogen or organic compounds function as the reductant1,2,5,11. Microbial involvement in U(VI) reduction has been considered to be limited to indirect effects, such as microbial metabolism providing the reduced compounds for abiological U(VI) reduction and microbial cell walls providing a surface to stimulate abiological U(VI) reduction1,12,13. We report here, however, that dissimilatory Fe(III)-reducing microorganisms can obtain energy for growth by electron transport to U(VI). This novel form of microbial metabolism can be much faster than commonly cited abiological mechanisms for U(VI) reduction. Not only do these findings expand the known potential terminal electron acceptors for microbial energy transduction, they offer a likely explanation for the deposition of uranium in aquatic sediments and aquifers, and suggest a method for biological remediation of environments contaminated with uranium.

  16. Reduction of astrometric plates

    NASA Technical Reports Server (NTRS)

    Stock, J.

    1984-01-01

    A rapid and accurate method for the reduction of comet or asteroid plates is described. Projection equations, scale length correction, rotation of coordinates, linearization, the search for additional reference stars, and the final solution are examined.

  17. Reduction of astrometric plates

    NASA Technical Reports Server (NTRS)

    Stock, J.

    1984-01-01

    A rapid and accurate method for the reduction of comet or asteroid plates is described. Projection equations, scale length correction, rotation of coordinates, linearization, the search for additional reference stars, and the final solution are examined.

  18. Waste Reduction Model

    EPA Pesticide Factsheets

    To help solid waste planners and organizations track/report GHG emissions reductions from various waste management practices. To assist in calculating GHG emissions of baseline and alternative waste management practices and provide the history of WARM.

  19. Reduction of bone strength

    NASA Technical Reports Server (NTRS)

    Bingham, Cindy

    1990-01-01

    Viewgraphs on reduction of bone strength are presented. WEHI 231 B growth rates, experimental chambers used to apply the electric field to the cell cultures, and a mouse suspended by rotating cuff in electromagnetic field are shown.

  20. Dissimilatory metal reduction.

    PubMed

    Lovley, D R

    1993-01-01

    Microorganisms can enzymatically reduce a variety of metals in metabolic processes that are not related to metal assimilation. Some microorganisms can conserve energy to support growth by coupling the oxidation of simple organic acids and alcohols, H2, or aromatic compounds to the reduction of Fe(III) or Mn(IV). This dissimilatory Fe(III) and Mn(IV) reduction influences the organic as well as the inorganic geochemistry of anaerobic aquatic sediments and ground water. Microorganisms that use U(VI) as a terminal electron acceptor play an important role in uranium geochemistry and may be a useful tool for removing uranium from contaminated environments. Se(VI) serves as a terminal electron acceptor to support anaerobic growth of some microorganisms. Reduction of Se(VI) to Se(O) is an important mechanism for the precipitation of selenium from contaminated waters. Enzymatic reduction of Cr(VI) to the less mobile and less toxic Cr(III), and reduction of soluble Hg(II) to volatile Hg(O) may affect the fate of these compounds in the environment and might be used as a remediation strategy. Microorganisms can also enzymatically reduce other metals such as technetium, vanadium, molybdenum, gold, silver, and copper, but reduction of these metals has not been studied extensively.

  1. The cyclic reduction algorithm

    NASA Astrophysics Data System (ADS)

    Bini, Dario; Meini, Beatrice

    2009-05-01

    Cyclic reduction is an algorithm invented by G.H. Golub and R. W. Hockney in the mid 1960s for solving linear systems related to the finite differences discretization of the Poisson equation over a rectangle. Among the algorithms of Gene Golub, it is one of the most versatile and powerful ever created. Recently, it has been applied to solve different problems from different applicative areas. In this paper we survey the main features of cyclic reduction, relate it to properties of analytic functions, recall its extension to solving more general finite and infinite linear systems, and different kinds of nonlinear matrix equations, including algebraic Riccati equations, with applications to Markov chains, queueing models and transport theory. Some new results concerning the convergence properties of cyclic reduction and its applicability are proved under very weak assumptions. New formulae for overcoming breakdown are provided.

  2. Time, Chance, and Reduction

    NASA Astrophysics Data System (ADS)

    Ernst, Gerhard; Hüttemann, Andreas

    2010-01-01

    List of contributors; 1. Introduction Gerhard Ernst and Andreas Hütteman; Part I. The Arrows of Time: 2. Does a low-entropy constraint prevent us from influencing the past? Mathias Frisch; 3. The part hypothesis meets gravity Craig Callender; 4. Quantum gravity and the arrow of time Claus Kiefer; Part II. Probability and Chance: 5. The natural-range conception of probability Jacob Rosenthal; 6. Probability in Boltzmannian statistical mechanics Roman Frigg; 7. Humean mechanics versus a metaphysics of powers Michael Esfeld; Part III. Reduction: 8. The crystallisation of Clausius's phenomenological thermodynamics C. Ulises Moulines; 9. Reduction and renormalization Robert W. Batterman; 10. Irreversibility in stochastic dynamics Jos Uffink; Index.

  3. REDUCTIONS WITHOUT REGRET: SUMMARY

    SciTech Connect

    Swegle, J.; Tincher, D.

    2013-09-16

    This paper briefly summarizes the series in which we consider the possibilities for losing, or compromising, key capabilities of the U.S. nuclear force in the face of modernization and reductions. The first of the three papers takes an historical perspective, considering capabilities that were eliminated in past force reductions. The second paper is our attempt to define the needed capabilities looking forward in the context of the current framework for force modernization and the current picture of the evolving challenges of deterrence and assurance. The third paper then provides an example for each of our undesirable outcomes: the creation of roach motels, box canyons, and wrong turns.

  4. Discrete reductive perturbation technique

    SciTech Connect

    Levi, Decio; Petrera, Matteo

    2006-04-15

    We expand a partial difference equation (P{delta}E) on multiple lattices and obtain the P{delta}E which governs its far field behavior. The perturbative-reductive approach is here performed on well-known nonlinear P{delta}Es, both integrable and nonintegrable. We study the cases of the lattice modified Korteweg-de Vries (mKdV) equation, the Hietarinta equation, the lattice Volterra-Kac-Van Moerbeke equation and a nonintegrable lattice KdV equation. Such reductions allow us to obtain many new P{delta}Es of the nonlinear Schroedinger type.

  5. Financing Class Size Reduction

    ERIC Educational Resources Information Center

    Achilles, C. M.

    2005-01-01

    Class size reduction has been shown to, among other things, improve academic achievement for all students and particularly for low-income and minority students. With the No Child Left Behind Act's heavy emphasis on scientifically based research, adequate yearly progress, and disaggregated results, one wonders why all children aren't enrolled in…

  6. Exercise and Fat Reduction.

    ERIC Educational Resources Information Center

    Clarke, H. Harrison, Ed.

    1975-01-01

    This document analyzes the problems encountered by the obese individual and the effects of regular exercise on weight loss and fat reduction. Part one compares the psychological traits of obese children with age groups of normal weight and discusses the organic disorders and social attitudes which plague the overweight individual. Part two states…

  7. Nagel on reduction.

    PubMed

    Sarkar, Sahotra

    2015-10-01

    This paper attempts a critical reappraisal of Nagel's (1961, 1970) model of reduction taking into account both traditional criticisms and recent defenses. This model treats reduction as a type of explanation in which a reduced theory is explained by a reducing theory after their relevant representational items have been suitably connected. In accordance with the deductive-nomological model, the explanation is supposed to consist of a logical deduction. Nagel was a pluralist about both the logical form of the connections between the reduced and reducing theories (which could be conditionals or biconditionals) and their epistemological status (as analytic connections, conventions, or synthetic claims). This paper defends Nagel's pluralism on both counts and, in the process, argues that the multiple realizability objection to reductionism is misplaced. It also argues that the Nagel model correctly characterizes reduction as a type of explanation. However, it notes that logical deduction must be replaced by a broader class of inferential techniques that allow for different types of approximation. Whereas Nagel (1970), in contrast to his earlier position (1961), recognized the relevance of approximation, he did not realize its full import for the model. Throughout the paper two case studies are used to illustrate the arguments: the putative reduction of classical thermodynamics to the kinetic theory of matter and that of classical genetics to molecular biology. Copyright © 2015. Published by Elsevier Ltd.

  8. Solar array cost reduction

    NASA Technical Reports Server (NTRS)

    Bernatowicz, D. T.

    1972-01-01

    A brief description is given of the cost of solar power systems over the last decade and means by which cost reductions may be achieved in the future. Costs were broken down into nonrecurring and recurring costs for solar array, battery, and power conditioning. Correlation of costs with power were poor; however, costs correlated reasonably well with the array area.

  9. Reduction in Force.

    ERIC Educational Resources Information Center

    Phay, Robert

    Chapter 2 in a book on school law discusses the reasons for reduction in force (RIF) and presents a set of model regulations for school districts as the best means of minimizing legal problems resulting from RIF. The reasons for RIF include declining student enrollments; reduced turnover among teachers; changes in programs; and more constrained…

  10. Simple Spectroscopy Reductions

    NASA Astrophysics Data System (ADS)

    Clayton, Martin; Holloway, Anthony

    This Cookbook describes the basic concepts and methods used in optical astronomical spectroscopy; it is aimed at those new to the field. Complete worked example reductions for both one- and two-dimensional longslit spectra, using real datasets, are described. Common problems and their solutions are discussed. A section on related resources is included, as is a glossary of commonly used terms.

  11. Parenteral packaging waste reduction.

    PubMed

    Baetz, B W

    1990-08-01

    The consumption of pharmaceutical products generates waste materials which can cause significant environmental impact when incinerated or landfilled. The purpose of this work is to stimulate discussion among hospital pharmacists and purchasing managers relating to the waste management aspects of their purchasing decisions. As a case study example, a number of commercially available "single use" parenterals are evaluated from a waste reduction perspective, for both the product container and for the packaging of these containers. Glass vials are non-incinerable, and are currently non-recyclable due to the higher melting temperatures required for borosilicate glass. However, plastic vials are potentially both incinerable and recyclable. Packaging quantities are considerably lower for plastic vials on a unit container basis, and also vary to a measurable degree between different manufacturers for a given type of container material. From an environmental perspective, waste reduction potential should become an important criterion in the selection of pharmaceutical products for hospital use.

  12. Injury reduction at Fermilab

    SciTech Connect

    Griffing, Bill; /Fermilab

    2005-06-01

    In a recent DOE Program Review, Fermilab's director presented results of the laboratory's effort to reduce the injury rate over the last decade. The results, shown in the figure below, reveal a consistent and dramatic downward trend in OSHA recordable injuries at Fermilab. The High Energy Physics Program Office has asked Fermilab to report in detail on how the laboratory has achieved the reduction. In fact, the reduction in the injury rate reflects a change in safety culture at Fermilab, which has evolved slowly over this period, due to a series of events, both planned and unplanned. This paper attempts to describe those significant events and analyze how each of them has shaped the safety culture that, in turn, has reduced the rate of injury at Fermilab to its current value.

  13. Fully Awake Breast Reduction.

    PubMed

    Filson, Simon A; Yarhi, Danielle; Ramon, Yitzhak

    2016-11-01

    The authors present 25 cases and an in-depth 4-minute video of fully awake aesthetic breast reduction, which was made possible by thoracic epidural anesthesia. There are obvious and important advantages to this technique. Not only does this allow for intraoperative patient cooperation (i.e., patient self-positioning and opinion for comparison of breasts), meaning a shorter and more efficient intraoperative time, there also is a reduction in postoperative pain, complications, recovery, and discharge times. The authors have also enjoyed great success and no complications with this technique in over 150 awake abdominoplasty/total body lift patients. The authors feel that the elimination of the need for general anesthesia by thoracic epidural sensorial-only anesthesia is a highly effective and efficient technique, with very few disadvantages/complications, providing advantages to both patients and surgeons. Therapeutic, IV.

  14. Aluminum reduction cell electrode

    DOEpatents

    Goodnow, Warren H.; Payne, John R.

    1982-01-01

    The invention is directed to cathode modules comprised of refractory hard metal materials, such as TiB.sub.2, for an electrolytic cell for the reduction of alumina wherein the modules may be installed and replaced during operation of the cell and wherein the structure of the cathode modules is such that the refractory hard metal materials are not subjected to externally applied forces or rigid constraints.

  15. Reduction of astrographic catalogues

    NASA Technical Reports Server (NTRS)

    Stock, J.; Prugna, F. D.; Cova, J.

    1984-01-01

    An automatic program for the reduction of overlapping Carte du Ciel plates is described. The projection and transformation equations are given and the RAA subprogram flow is outlined. The program was applied to two different sets of data, namely to nine overlapping plates of the Cape Zone of the CdC, and to fifteen plates taken with the CIDA-refractor of the open cluster Tr10.

  16. Somatic reduction in cycads.

    PubMed

    Storey, W B

    1968-02-09

    Recurrent somatic reduction is a normal ontogenetic process in apogeotropic roots of cycads, which develop into dichotomously branching coralloid masses. The reduced cells make up part of a ring of differentiated cortical tissue lying midway between the pericycle and the epidermis; they serve as fillers among the large cells and become charged with slime. The differentiated tissue is colonized by a species of blue-green algae.

  17. Drag reduction strategies

    NASA Technical Reports Server (NTRS)

    Hill, D. Christopher

    1994-01-01

    previously a description was given of an active control scheme using wall transpiration that leads to a 15% reduction in surface skin friction beneath a turbulent boundary layer, according to direct numerical simulation. In this research brief further details of that scheme and its variants are given together with some suggestions as to how sensor/actuator arrays could be configured to reduce surface drag. The research which is summarized here was performed during the first half of 1994. This research is motivated by the need to understand better how the dynamics of near-wall turbulent flow can be modified so that skin friction is reduced. The reduction of turbulent skin friction is highly desirable in many engineering applications. Experiments and direct numerical simulations have led to an increased understanding of the cycle of turbulence production and transport in the boundary layer and raised awareness of the possibility of disrupting the process with a subsequent reduction in turbulent skin friction. The implementation of active feedback control in a computational setting is a viable approach for the investigation of the modifications to the flow physics that can be achieved. Bewley et al. and Hill describe how ideas from optimal control theory are employed to give 'sub-optimal' drag reduction schemes. The objectives of the work reported here is to investigate in greater detail the assumptions implicit within such schemes and their limitations. It is also our objective to describe how an array of sensors and actuators could be arranged and interconnected to form a 'smart' surface which has low skin friction.

  18. Reduction of astrographic catalogues

    NASA Technical Reports Server (NTRS)

    Stock, J.; Prugna, F. D.; Cova, J.

    1984-01-01

    An automatic program for the reduction of overlapping Carte du Ciel plates is described. The projection and transformation equations are given and the RAA subprogram flow is outlined. The program was applied to two different sets of data, namely to nine overlapping plates of the Cape Zone of the CdC, and to fifteen plates taken with the CIDA-refractor of the open cluster Tr10.

  19. Oxidation, Reduction, and Deoxygenation

    NASA Astrophysics Data System (ADS)

    Madsen, Robert

    In this chapter, methods for oxidation, reduction, and deoxygenation of carbohydrates are presented. In most cases, the reactions have been used on aldoses and their derivatives including glycosides, uronic acids, glycals, and other unsaturated monosaccharides. A number of reactions have also been applied to aldonolactones. The methods include both chemical and enzymatic procedures and some of these can be applied for regioselective transformation of unprotected or partially protected carbohydrates.

  20. Television noise reduction device

    NASA Technical Reports Server (NTRS)

    Gordon, B. L.; Stamps, J. C. (Inventor)

    1975-01-01

    A noise reduction system that divides the color video signal into its luminance and chrominance components is reported. The luminance component of a given frame is summed with the luminance component of at least one preceding frame which was stored on a disc recorder. The summation is carried out so as to achieve a signal amplitude equivalent to that of the original signal. The averaged luminance signal is then recombined with the chrominance signal to achieve a noise-reduced television signal.

  1. Aluminum reduction cell electrode

    DOEpatents

    Goodnow, W.H.; Payne, J.R.

    1982-09-14

    The invention is directed to cathode modules comprised of refractory hard metal materials, such as TiB[sub 2], for an electrolytic cell for the reduction of alumina wherein the modules may be installed and replaced during operation of the cell and wherein the structure of the cathode modules is such that the refractory hard metal materials are not subjected to externally applied forces or rigid constraints. 9 figs.

  2. Uranium Reduction by Clostridia

    SciTech Connect

    Francis, A.J.; Dodge, Cleveland J.; Gillow, Jeffrey B.

    2006-04-05

    The FRC groundwater and sediment contain significant concentrations of U and Tc and are dominated by low pH, and high nitrate and Al concentrations where dissimilatory metal reducing bacterial activity may be limited. The presence of Clostridia in Area 3 at the FRC site has been confirmed and their ability to reduce uranium under site conditions will be determined. Although the phenomenon of uranium reduction by Clostridia has been firmly established, the molecular mechanisms underlying such a reaction are not very clear. The authors are exploring the hypothesis that U(VI) reduction occurs through hydrogenases and other enzymes (Matin and Francis). Fundamental knowledge of metal reduction using Clostridia will allow us to exploit naturally occurring processes to attenuate radionuclide and metal contaminants in situ in the subsurface. The outline for this report are as follows: (1) Growth of Clostridium sp. under normal culture conditions; (2) Fate of metals and radionuclides in the presence of Clostridia; (3) Bioreduction of uranium associated with nitrate, citrate, and lepidocrocite; and (4) Utilization of Clostridium sp. for immobilization of uranium at the FRC Area 3 site.

  3. Thermochemical nitrate reduction

    SciTech Connect

    Cox, J.L.; Lilga, M.A.; Hallen, R.T.

    1992-09-01

    A series of preliminary experiments was conducted directed at thermochemically converting nitrate to nitrogen and water. Nitrates are a major constituent of the waste stored in the underground tanks on the Hanford Site, and the characteristics and effects of nitrate compounds on stabilization techniques must be considered before permanent disposal operations begin. For the thermochemical reduction experiments, six reducing agents (ammonia, formate, urea, glucose, methane, and hydrogen) were mixed separately with {approximately}3 wt% NO{sub 3}{sup {minus}} solutions in a buffered aqueous solution at high pH (13); ammonia and formate were also mixed at low pH (4). Reactions were conducted in an aqueous solution in a batch reactor at temperatures of 200{degrees}C to 350{degrees}C and pressures of 600 to 2800 psig. Both gas and liquid samples were analyzed. The specific components analyzed were nitrate, nitrite, nitrous oxide, nitrogen, and ammonia. Results of experimental runs showed the following order of nitrate reduction of the six reducing agents in basic solution: formate > glucose > urea > hydrogen > ammonia {approx} methane. Airnmonia was more effective under acidic conditions than basic conditions. Formate was also effective under acidic conditions. A more thorough, fundamental study appears warranted to provide additional data on the mechanism of nitrate reduction. Furthermore, an expanded data base and engineering feasibility study could be used to evaluate conversion conditions for promising reducing agents in more detail and identify new reducing agents with improved performance characteristics.

  4. Reduction of turbomachinery noise

    NASA Technical Reports Server (NTRS)

    Waitz, Ian A. (Inventor); Brookfield, John M. (Inventor); Sell, Julian (Inventor); Hayden, Belva J. (Inventor); Ingard, K. Uno (Inventor)

    1999-01-01

    In the invention, propagating broad band and tonal acoustic components of noise characteristic of interaction of a turbomachine blade wake, produced by a turbomachine blade as the blade rotates, with a turbomachine component downstream of the rotating blade, are reduced. This is accomplished by injection of fluid into the blade wake through a port in the rotor blade. The mass flow rate of the fluid injected into the blade wake is selected to reduce the momentum deficit of the wake to correspondingly increase the time-mean velocity of the wake and decrease the turbulent velocity fluctuations of the wake. With this fluid injection, reduction of both propagating broad band and tonal acoustic components of noise produced by interaction of the blade wake with a turbomachine component downstream of the rotating blade is achieved. In a further noise reduction technique, boundary layer fluid is suctioned into the turbomachine blade through a suction port on the side of the blade that is characterized as the relatively low-pressure blade side. As with the fluid injection technique, the mass flow rate of the fluid suctioned into the blade is here selected to reduce the momentum deficit of the wake to correspondingly increase the time-mean velocity of the wake and decrease the turbulent velocity fluctuations of the wake; reduction of both propagating broad band and tonal acoustic components of noise produced by interaction of the blade wake with a turbomachine component downstream of the rotating blade is achieved with this suction technique. Blowing and suction techniques are also provided in the invention for reducing noise associated with the wake produced by fluid flow around a stationary blade upstream of a rotating turbomachine.

  5. Aluminum reduction cell electrode

    DOEpatents

    Payne, John R.

    1983-09-20

    The invention is directed to an anode-cathode structure for an electrolytic cell for the reduction of alumina wherein the structure is comprised of a carbon anode assembly which straddles a wedge-shaped refractory hard metal cathode assembly having steeply sloped cathodic surfaces, each cathodic surface being paired in essentially parallel planar relationship with an anode surface. The anode-cathode structure not only takes into account the structural weakness of refractory hard metal materials but also permits the changing of the RHM assembly during operation of the cell. Further, the anode-cathode structure enhances the removal of anode gas from the interpolar gap between the anode and cathode surfaces.

  6. Electrolytic oxide reduction system

    DOEpatents

    Wiedmeyer, Stanley G; Barnes, Laurel A; Williamson, Mark A; Willit, James L; Berger, John F

    2015-04-28

    An electrolytic oxide reduction system according to a non-limiting embodiment of the present invention may include a plurality of anode assemblies, a plurality of cathode assemblies, and a lift system configured to engage the anode and cathode assemblies. The cathode assemblies may be alternately arranged with the anode assemblies such that each cathode assembly is flanked by two anode assemblies. The lift system may be configured to selectively engage the anode and cathode assemblies so as to allow the simultaneous lifting of any combination of the anode and cathode assemblies (whether adjacent or non-adjacent).

  7. Aircraft engine pollution reduction

    NASA Technical Reports Server (NTRS)

    Rudey, R. A.

    1972-01-01

    The effect of engine operation on the types and levels of the major aircraft engine pollutants is described and the major factors governing the formation of these pollutants during the burning of hydrocarbon fuel are discussed. Methods which are being explored to reduce these pollutants are discussed and their application to several experimental research programs are pointed out. Results showing significant reductions in the levels of carbon monoxide, unburned hydrocarbons, and oxides of nitrogen obtained from experimental combustion research programs are presented and discussed to point out potential application to aircraft engines.

  8. NSF grant reductions

    NASA Astrophysics Data System (ADS)

    Jones, R.

    Many National Science Foundation grants will be reduced this year as a result of a provision in H.R. 3299. The provision stems from disagreement between the Congress and the administration on how to make budget deficit cuts required by the Gramm-Rudman-Hollings budget law. An agreement was made to cut $4.55 billion through a reduction in discretionary spending, by what amounts to 1.4% across-the-board. The cuts will affect all discretionary federal domestic and defense programs.

  9. Reduction operators of Burgers equation

    PubMed Central

    Pocheketa, Oleksandr A.; Popovych, Roman O.

    2013-01-01

    The solution of the problem on reduction operators and nonclassical reductions of the Burgers equation is systematically treated and completed. A new proof of the theorem on the special “no-go” case of regular reduction operators is presented, and the representation of the coefficients of operators in terms of solutions of the initial equation is constructed for this case. All possible nonclassical reductions of the Burgers equation to single ordinary differential equations are exhaustively described. Any Lie reduction of the Burgers equation proves to be equivalent via the Hopf–Cole transformation to a parameterized family of Lie reductions of the linear heat equation. PMID:23576819

  10. Reduction operators of Burgers equation.

    PubMed

    Pocheketa, Oleksandr A; Popovych, Roman O

    2013-02-01

    The solution of the problem on reduction operators and nonclassical reductions of the Burgers equation is systematically treated and completed. A new proof of the theorem on the special "no-go" case of regular reduction operators is presented, and the representation of the coefficients of operators in terms of solutions of the initial equation is constructed for this case. All possible nonclassical reductions of the Burgers equation to single ordinary differential equations are exhaustively described. Any Lie reduction of the Burgers equation proves to be equivalent via the Hopf-Cole transformation to a parameterized family of Lie reductions of the linear heat equation.

  11. Microbial reduction of iodate

    USGS Publications Warehouse

    Councell, T.B.; Landa, E.R.; Lovley, D.R.

    1997-01-01

    The different oxidation species of iodine have markedly different sorption properties. Hence, changes in iodine redox states can greatly affect the mobility of iodine in the environment. Although a major microbial role has been suggested in the past to account for these redox changes, little has been done to elucidate the responsible microorganisms or the mechanisms involved. In the work presented here, direct microbial reduction of iodate was demonstrated with anaerobic cell suspensions of the sulfate reducing bacterium Desulfovibrio desulfuricans which reduced 96% of an initial 100 ??M iodate to iodide at pH 7 in 30 mM NaHCO3 buffer, whereas anaerobic cell suspensions of the dissimilatory Fe(III)-reducing bacterium Shewanella putrefaciens were unable to reduce iodate in 30 mM NaHCO3 buffer (pH 7). Both D. desulfuricans and S. putrefaciens were able to reduce iodate at pH 7 in 10 mM HEPES buffer. Both soluble ferrous iron and sulfide, as well as iron monosulfide (FeS) were shown to abiologically reduce iodate to iodide. These results indicate that ferric iron and/or sulfate reducing bacteria are capable of mediating both direct, enzymatic, as well as abiotic reduction of iodate in natural anaerobic environments. These microbially mediated reactions may be important factors in the fate and transport of 129I in natural systems.

  12. Core Noise Reduction

    NASA Technical Reports Server (NTRS)

    Hultgren, Lennart S.

    2011-01-01

    This presentation is a technical summary of and outlook for NASA-internal and NASA-sponsored external research on core (combustor and turbine) noise funded by the Fundamental Aeronautics Program Subsonic Fixed Wing (SFW) Project. Sections of the presentation cover: the SFW system-level noise metrics for the 2015, 2020, and 2025 timeframes; turbofan design trends and their aeroacoustic implications; the emerging importance of core noise and its relevance to the SFW Reduce-Perceived-Noise Technical Challenge; and the current research activities in the core noise area. Recent work1 on the turbine-transmission loss of combustor noise is briefly described, two2,3 new NRA efforts in the core-noise area are outlined, and an effort to develop CMC-based acoustic liners for broadband noise reduction suitable for turbofan-core application is delineated. The NASA Fundamental Aeronautics Program has the principal objective of overcoming today's national challenges in air transportation. The reduction of aircraft noise is critical to enabling the anticipated large increase in future air traffic. The Subsonic Fixed Wing Project's Reduce-Perceived-Noise Technical Challenge aims to develop concepts and technologies to dramatically reduce the perceived aircraft noise outside of airport boundaries.

  13. UCAC3: ASTROMETRIC REDUCTIONS

    SciTech Connect

    Finch, Charlie T.; Zacharias, Norbert; Wycoff, Gary L.

    2010-06-15

    Presented here are the details of the astrometric reductions from the x, y data to mean right ascension (R.A.), declination (decl.) coordinates of the third U.S. Naval Observatory CCD Astrograph Catalog (UCAC3). For these new reductions we used over 216,000 CCD exposures. The Two-Micron All-Sky Survey (2MASS) data are used extensively to probe for coordinate and coma-like systematic errors in UCAC data mainly caused by the poor charge transfer efficiency of the 4K CCD. Errors up to about 200 mas have been corrected using complex look-up tables handling multiple dependences derived from the residuals. Similarly, field distortions and sub-pixel phase errors have also been evaluated using the residuals with respect to 2MASS. The overall magnitude equation is derived from UCAC calibration field observations alone, independent of external catalogs. Systematic errors of positions at the UCAC observing epoch as presented in UCAC3 are better corrected than in the previous catalogs for most stars. The Tycho-2 catalog is used to obtain final positions on the International Celestial Reference Frame. Residuals of the Tycho-2 reference stars show a small magnitude equation (depending on declination zone) that might be inherent in the Tycho-2 catalog.

  14. Harm Reduction From Below

    PubMed Central

    Van Schipstal, Inge; Berning, Moritz; Murray, Hayley

    2016-01-01

    This article focuses on how recreational drug users in the Netherlands and in online communities navigate the risks and reduce the harms they associate with psychoactive drug use. To do so, we examined the protective practices they invent, use, and share with their immediate peers and with larger drug experimenting communities online. The labor involved in protective practices and that which ultimately informs harm reduction from below follows three interrelated trajectories: (1) the handling and sharing of drugs to facilitate hassle-free drug use, (2) creating pleasant and friendly spaces that we highlight under the practices of drug use attunements, and (3) the seeking and sharing of information in practices to spread the good high. We focus not only on users’ concerns but also on how these concerns shape their approach to drugs, what young people do to navigate uncertainties, and how they reach out to and create different sources of knowledge to minimize adversities and to improve highs. Harm reduction from below, we argue, can best be seen in the practices of sharing around drug use and in the caring for the larger community of drug-using peers. PMID:27721525

  15. Islam and harm reduction.

    PubMed

    Kamarulzaman, A; Saifuddeen, S M

    2010-03-01

    Although drugs are haram and therefore prohibited in Islam, illicit drug use is widespread in many Islamic countries throughout the world. In the last several years increased prevalence of this problem has been observed in many of these countries which has in turn led to increasing injecting drug use driven HIV/AIDS epidemic across the Islamic world. Whilst some countries have recently responded to the threat through the implementation of harm reduction programmes, many others have been slow to respond. In Islam, The Quran and the Prophetic traditions or the Sunnah are the central sources of references for the laws and principles that guide the Muslims' way of life and by which policies and guidelines for responses including that of contemporary social and health problems can be derived. The preservation and protection of the dignity of man, and steering mankind away from harm and destruction are central to the teachings of Islam. When viewed through the Islamic principles of the preservation and protection of the faith, life, intellect, progeny and wealth, harm reduction programmes are permissible and in fact provide a practical solution to a problem that could result in far greater damage to the society at large if left unaddressed. Copyright (c) 2009. Published by Elsevier B.V.

  16. Sonoassisted microbial reduction of chromium.

    PubMed

    Kathiravan, Mathur Nadarajan; Karthick, Ramalingam; Muthu, Naggapan; Muthukumar, Karuppan; Velan, Manickam

    2010-04-01

    This study presents sonoassisted microbial reduction of hexavalent chromium (Cr(VI)) using Bacillus sp. isolated from tannery effluent contaminated site. The experiments were carried out with free cells in the presence and absence of ultrasound. The optimum pH and temperature for the reduction of Cr(VI) by Bacillus sp. were found to be 7.0 and 37 degrees C, respectively. The Cr(VI) reduction was significantly influenced by the electron donors and among the various electron donors studied, glucose offered maximum reduction. The ultrasound-irradiated reduction of Cr(VI) with Bacillus sp. showed efficient Cr(VI) reduction. The percent reduction was found to increase with an increase in biomass concentration and decrease with an increase in initial concentration. The changes in the functional groups of Bacillus sp., before and after chromium reduction were observed with FTIR spectra. Microbial growth was described with Monod and Andrews model and best fit was observed with Andrews model.

  17. Tritium Emissions Reduction Facility

    SciTech Connect

    Wieneke, R.E.; Bowser, R.P.; Hedley, W.H.; Kissner, T.J.; Lamberger, P.H.; Morgan, F.G.; Van Patten, J.F.; Williams, M.A.

    1988-01-01

    The Tritium Emissions Reduction Facility (TERF) will be a system for the continuous processing of tritium containing gases collected from various operations at Mound. The basis of the system operation will be the oxidation of elemental hydrogen isotopes and organic molecules at elevated temperatures on precious metal catalyst beds, and the adsorption of the resulting oxide (water) on molecular sieve dryers. The TERF will be expected to handle from 400,000 to 1,000,000 curies of tritium per year in the process gas stream and release no more than 200 curies per year to the atmosphere. Consequently, the TERF will need to convert and capture tritium at low concentrations in gas efficiently and reliably. 5 refs., 2 figs.

  18. Aircraft engine pollution reduction.

    NASA Technical Reports Server (NTRS)

    Rudey, R. A.

    1972-01-01

    The effect of engine operation on the types and levels of the major aircraft engine pollutants is described and the major factors governing the formation of these pollutants during the burning of hydrocarbon fuel are discussed. Methods which are being explored to reduce these pollutants are discussed and their application to several experimental research programs are pointed out. Results showing significant reductions in the levels of carbon monoxide, unburned hydrocarbons, and oxides of nitrogen obtained from experimental combustion research programs are presented and discussed to point out potential application to aircraft engines. An experimental program designed to develop and demonstrate these and other advanced, low pollution combustor design methods is described. Results that have been obtained to date indicate considerable promise for reducing advanced engine exhaust pollutants to levels significantly below current engines.

  19. Active noise reduction

    NASA Astrophysics Data System (ADS)

    Geyer, Carolyn R.

    Active noise reduction (ANR) techniques are described with reference to their application to crewmembers during aircraft operation to enhance productivity and safety. ANR concepts and theory are explained, and the development of protective ANR systems for direct implementation are described. Sound attenuation testing was conducted to study the feasibility of aircraft-powered ANR systems, and the positive results spurred their development for compatibility with flight helmets. The Helmets Limited ANR system uses a bypass mode at times of limited available power and complements the use of passive sound attenuation. Subjective testing results show that the device is effective, and a planned program of intensive evaluation is discussed. The aircraft that require an ANR system are listed, and key areas of implementation include battery power and the combination of ANR circuitry and helmet oxygen masks. It is suggested that ANR techniques can positively impact the efficiency and performance of crewmembers in high-noise-level aircraft.

  20. Reduction of polysymplectic manifolds

    NASA Astrophysics Data System (ADS)

    Marrero, Juan Carlos; Román-Roy, Narciso; Salgado, Modesto; Vilariño, Silvia

    2015-02-01

    The aim of this paper is to generalize the classical Marsden-Weinstein reduction procedure for symplectic manifolds to polysymplectic manifolds in order to obtain quotient manifolds which inherit the polysymplectic structure. This generalization allows us to reduce polysymplectic Hamiltonian systems with symmetries, such as those appearing in certain kinds of classical field theories. As an application of this technique, an analogue to the Kirillov-Kostant-Souriau theorem for polysymplectic manifolds is obtained and some other mathematical examples are also analyzed. Our procedure corrects some mistakes and inaccuracies in previous papers (Günther 1987 J. Differ. Geom. 25 23-53 Munteanu et al 2004 J. Math. Phys. 45 1730-51) on this subject.

  1. Aluminum reduction cell electrode

    DOEpatents

    Payne, J.R.

    1983-09-20

    The invention is directed to an anode-cathode structure for an electrolytic cell for the reduction of alumina wherein the structure is comprised of a carbon anode assembly which straddles a wedge-shaped refractory hard metal cathode assembly having steeply sloped cathodic surfaces, each cathodic surface being paired in essentially parallel planar relationship with an anode surface. The anode-cathode structure not only takes into account the structural weakness of refractory hard metal materials but also permits the changing of the RHM assembly during operation of the cell. Further, the anode-cathode structure enhances the removal of anode gas from the interpolar gap between the anode and cathode surfaces. 10 figs.

  2. Final reduction gear apparatus

    SciTech Connect

    Yasui, Y.; Hori, H.

    1987-04-21

    A final reduction gear apparatus is described comprising: a differential carrier which houses a gear assembly; an oil seal attached to a side gear shaft opening in the differential carrier, the oil seal having a main lip which may contact a periphery of a side gear shaft; and a guide member located outside of the oil seal at the side gear shaft opening, the guide member being formed as a member separate from the oil seal, the guide member having a slightly larger inner diameter than that of the main lip of the oil seal, and having guide surface concentric to the main lip, wherein 1/2 of the difference between the inner diameter of the guide member and the inner diameter of the main lip of the oil seal is within the limit of the elastic deformability of the main lip.

  3. Dose Reduction Techniques

    SciTech Connect

    WAGGONER, L.O.

    2000-05-16

    As radiation safety specialists, one of the things we are required to do is evaluate tools, equipment, materials and work practices and decide whether the use of these products or work practices will reduce radiation dose or risk to the environment. There is a tendency for many workers that work with radioactive material to accomplish radiological work the same way they have always done it rather than look for new technology or change their work practices. New technology is being developed all the time that can make radiological work easier and result in less radiation dose to the worker or reduce the possibility that contamination will be spread to the environment. As we discuss the various tools and techniques that reduce radiation dose, keep in mind that the radiological controls should be reasonable. We can not always get the dose to zero, so we must try to accomplish the work efficiently and cost-effectively. There are times we may have to accept there is only so much you can do. The goal is to do the smart things that protect the worker but do not hinder him while the task is being accomplished. In addition, we should not demand that large amounts of money be spent for equipment that has marginal value in order to save a few millirem. We have broken the handout into sections that should simplify the presentation. Time, distance, shielding, and source reduction are methods used to reduce dose and are covered in Part I on work execution. We then look at operational considerations, radiological design parameters, and discuss the characteristics of personnel who deal with ALARA. This handout should give you an overview of what it takes to have an effective dose reduction program.

  4. Reductant injection and mixing system

    SciTech Connect

    Reeves, Matt; Henry, Cary A.; Ruth, Michael J.

    2016-02-16

    A gaseous reductant injection and mixing system is described herein. The system includes an injector for injecting a gaseous reductant into an exhaust gas stream, and a mixer attached to a surface of the injector. The injector includes a plurality of apertures through which the gaseous reductant is injected into an exhaust gas stream. The mixer includes a plurality of fluid deflecting elements.

  5. Jet Noise Reduction

    NASA Technical Reports Server (NTRS)

    Kenny, Patrick

    2004-01-01

    The Acoustics Branch is responsible for reducing noise levels for jet and fan components on aircraft engines. To do this, data must be measured and calibrated accurately to ensure validity of test results. This noise reduction is accomplished by modifications to hardware such as jet nozzles, and by the use of other experimental hardware such as fluidic chevrons, elliptic cores, and fluidic shields. To insure validity of data calibration, a variety of software is used. This software adjusts the sound amplitude and frequency to be consistent with data taken on another day. Both the software and the hardware help make noise reduction possible. work properly. These software programs were designed to make corrections for atmosphere, shear, attenuation, electronic, and background noise. All data can be converted to a one-foot lossless condition, using the proper software corrections, making a reading independent of weather and distance. Also, data can be transformed from model scale to full scale for noise predictions of a real flight. Other programs included calculations of Over All Sound Pressure Level (OASPL), Effective Perceived Noise Level (EPNL). OASPL is the integration of sound with respect to frequency, and EPNL is weighted for a human s response to different sound frequencies and integrated with respect to time. With the proper software correction, data taken in the NATR are useful in determining ways to reduce noise. display any difference between two or more data files. Using this program and graphs of the data, the actual and predicted data can be compared. This software was tested on data collected at the Aero Acoustic Propulsion Laboratory (AAPL) using a variety of window types and overlaps. Similarly, short scripts were written to test each individual program in the software suite for verification. Each graph displays both the original points and the adjusted points connected with lines. During this summer, data points were taken during a live experiment

  6. Size reduction machine

    SciTech Connect

    Fricke, V.

    1999-12-15

    The Size Reduction Machine (SRM) is a mobile platform capable of shearing various shapes and types of metal components at a variety of elevations. This shearing activity can be performed without direct physical movement and placement of the shear head by the operator. The base unit is manually moved and roughly aligned to each cut location. The base contains the electronics: hydraulic pumps, servos, and actuators needed to move the shear-positioning arm. The movable arm allows the shear head to have six axes of movement and to cut to within 4 inches of a wall surface. The unit has a slick electrostatic capture coating to assist in external decontamination. Internal contamination of the unit is controlled by a high-efficiency particulate air (HEPA) filter on the cooling inlet fan. The unit is compact enough to access areas through a 36-inch standard door opening. This paper is an Innovative Technology Summary Report designed to provide potential users with the information they need to quickly determine if a technology would apply to a particular environmental management problem. They also are designed for readers who may recommend that a technology be considered by prospective users.

  7. Heart Failure Readmission Reduction.

    PubMed

    Drozda, Joseph P; Smith, Donna A; Freiman, Paul C; Pursley, Janet; VanSlette, Jeffrey A; Smith, Timothy R

    Little is known regarding effectiveness of readmission reduction programs over time. The Heart Failure Management Program (HFMP) of St. John's Physician Group Practice (PGP) Demonstration provided an opportunity to assess outcomes over an extended period. Data from an electronic health record, an inpatient database, a disease registry, and the Social Security Death Master File were analyzed for patients admitted with heart failure (HF) for 5 years before (Period 1) and 5 years after (Period 2) inception of PGP. HF admissions decreased (Period 1, 58.3/month; Period 2, 52.4/month, P = .007). Thirty-day all-cause readmission rate dropped from Period 1 (annual average 18.8% [668/3545]) to year 1 of Period 2 (16.9% [136/804], P = .04) and remained stable thereafter (annual average 16.8% [589/3503]). Thirty-day mortality rate was flat throughout. HFMP was associated with decreased readmissions, primarily related to outpatient case management, while mortality remained stable.

  8. Smokeless Tobacco Reduction Program.

    PubMed

    Glidden, L; Whigam, K

    1987-01-01

    To reduce the incidence and prevalence of oral cancer, the Smokeless Tobacco Reduction Program will consist of a mass media campaign, public oral screening, and a week-long school health program for 350 students in the seventh, eighth, and ninth grades in Willows, Glenn County, CA. Mass media will include radio, television, newspapers, posters, and literature. The program will use resources of the public health department and junior high school; it will also depend on 8 teachers and 25 peer leaders, all trained in the program. Reducing the use of smokeless tobacco is the program's objective. If that goal is achieved, the program will reduce oral cancers in the target population by 75 percent within 10 years. The incidence of leukoplakia will be reduced by 50 percent within 3 years of the end of the program. By the end of the program, 90 percent of the target population will be able to identify warning signs of oral cancer and leukoplakia, and 85 percent of the students will no longer believe that use of smokeless tobacco is less harmful than smoking. As a result of the program, use of smokeless tobacco will not be viewed favorably by 80 percent of the target population; usage will be regarded as socially unacceptable.

  9. Principal Components as a Data Reduction and Noise Reduction Technique

    NASA Technical Reports Server (NTRS)

    Imhoff, M. L.; Campbell, W. J.

    1982-01-01

    The potential of principal components as a pipeline data reduction technique for thematic mapper data was assessed and principal components analysis and its transformation as a noise reduction technique was examined. Two primary factors were considered: (1) how might data reduction and noise reduction using the principal components transformation affect the extraction of accurate spectral classifications; and (2) what are the real savings in terms of computer processing and storage costs of using reduced data over the full 7-band TM complement. An area in central Pennsylvania was chosen for a study area. The image data for the project were collected using the Earth Resources Laboratory's thematic mapper simulator (TMS) instrument.

  10. Model reduction for flexible structures

    NASA Technical Reports Server (NTRS)

    Gawronski, Wodek; Juang, Jer-Nan

    1990-01-01

    Several conditions for a near-optimal reduction of general dynamic systems are presented focusing on the reduction in balanced and modal coordinates. It is shown that model and balanced reductions give very different results for the flexible structure with closely-spaced natural frequencies. In general, balanced reduction is found to give better results. A robust model reduction technique was developed to study the sensitivity of modeling error to variations in the damping of a structure. New concepts of grammians defined over a finite time and/or a frequency interval are proposed including computational procedures for evaluating them. Application of the model reduction technique to these grammians is considered to lead to a near-optimal reduced model which closely reproduces the full system output in the time and/or frequency interval.

  11. Emerging Community Noise Reduction Approaches

    NASA Technical Reports Server (NTRS)

    Envia, Edmane

    2012-01-01

    An overview of the current NASA research portfolio in the area of aircraft noise reduction is presented. The emphasis of the research described herein is on meeting the aggressive near- and mid-term national goals for reducing aircraft noise emissions, which NASA internal studies have shown to be feasible using noise reduction technologies currently being developed in-house or in partnership with NASA s industry and academic partners. While NASA has an active research effort in airframe noise reduction, this overview focuses on propulsion noise reduction only.

  12. Routh reduction and Cartan mechanics

    NASA Astrophysics Data System (ADS)

    Capriotti, S.

    2017-04-01

    In the present work a Cartan mechanics version for Routh reduction is considered, as an intermediate step towards Routh reduction in field theory. Motivation for this generalization comes from a scheme for integrable systems (Fehér and Gábor, 2002), used for understanding the occurrence of Toda field theories in so called Hamiltonian reduction of WZNW field theories (Fehér et al., 1992). As a way to accomplish with this intermediate aim, this article also contains a formulation of the Lagrangian Adler-Kostant-Symes systems discussed in Fehér and Gábor (2002) in terms of Routh reduction.

  13. Fraction Reduction in Membrane Systems

    PubMed Central

    Zhang, Hong

    2014-01-01

    Fraction reduction is a basic computation for rational numbers. P system is a new computing model, while the current methods for fraction reductions are not available in these systems. In this paper, we propose a method of fraction reduction and discuss how to carry it out in cell-like P systems with the membrane structure and the rules with priority designed. During the application of fraction reduction rules, synchronization is guaranteed by arranging some special objects in these rules. Our work contributes to performing the rational computation in P systems since the rational operands can be given in the form of fraction. PMID:24772037

  14. Cochlear implant optimized noise reduction

    NASA Astrophysics Data System (ADS)

    Mauger, Stefan J.; Arora, Komal; Dawson, Pam W.

    2012-12-01

    Noise-reduction methods have provided significant improvements in speech perception for cochlear implant recipients, where only quality improvements have been found in hearing aid recipients. Recent psychoacoustic studies have suggested changes to noise-reduction techniques specifically for cochlear implants, due to differences between hearing aid recipient and cochlear implant recipient hearing. An optimized noise-reduction method was developed with significantly increased temporal smoothing of the signal-to-noise ratio estimate and a more aggressive gain function compared to current noise-reduction methods. This optimized noise-reduction algorithm was tested with 12 cochlear implant recipients over four test sessions. Speech perception was assessed through speech in noise tests with three noise types; speech-weighted noise, 20-talker babble and 4-talker babble. A significant speech perception improvement using optimized noise reduction over standard processing was found in babble noise and speech-weighted noise and over a current noise-reduction method in speech-weighted noise. Speech perception in quiet was not degraded. Listening quality testing for noise annoyance and overall preference found significant improvements over the standard processing and over a current noise-reduction method in speech-weighted and babble noise types. This optimized method has shown significant speech perception and quality improvements compared to the standard processing and a current noise-reduction method.

  15. Cochlear implant optimized noise reduction.

    PubMed

    Mauger, Stefan J; Arora, Komal; Dawson, Pam W

    2012-12-01

    Noise-reduction methods have provided significant improvements in speech perception for cochlear implant recipients, where only quality improvements have been found in hearing aid recipients. Recent psychoacoustic studies have suggested changes to noise-reduction techniques specifically for cochlear implants, due to differences between hearing aid recipient and cochlear implant recipient hearing. An optimized noise-reduction method was developed with significantly increased temporal smoothing of the signal-to-noise ratio estimate and a more aggressive gain function compared to current noise-reduction methods. This optimized noise-reduction algorithm was tested with 12 cochlear implant recipients over four test sessions. Speech perception was assessed through speech in noise tests with three noise types; speech-weighted noise, 20-talker babble and 4-talker babble. A significant speech perception improvement using optimized noise reduction over standard processing was found in babble noise and speech-weighted noise and over a current noise-reduction method in speech-weighted noise. Speech perception in quiet was not degraded. Listening quality testing for noise annoyance and overall preference found significant improvements over the standard processing and over a current noise-reduction method in speech-weighted and babble noise types. This optimized method has shown significant speech perception and quality improvements compared to the standard processing and a current noise-reduction method.

  16. Noise Reduction by Signal Accumulation

    ERIC Educational Resources Information Center

    Kraftmakher, Yaakov

    2006-01-01

    The aim of this paper is to show how the noise reduction by signal accumulation can be accomplished with a data acquisition system. This topic can be used for student projects. In many cases, the noise reduction is an unavoidable part of experimentation. Several techniques are known for this purpose, and among them the signal accumulation is the…

  17. Aircraft Noise Reduction Subproject Overview

    NASA Technical Reports Server (NTRS)

    Fernandez, Hamilton; Nark, Douglas M.; Van Zante, Dale E.

    2016-01-01

    The material presents highlights of propulsion and airframe noise research being completed for the Advanced Air Transport Technology Project. The basis of noise reduction plans along with representative work for the airframe, propulsion, and propulsion-airframe integration is discussed for the Aircraft Noise reduction Subproject.

  18. Reduction-Fired Seedpod Bowls.

    ERIC Educational Resources Information Center

    Beyke, Rod

    2001-01-01

    Focuses on a reduction-firing process with an aim of producing high-quality blackware similar to the black-on-black pottery of Maria Martinez and other American Indian potters. Includes a lesson on creating reduction-fired seedpod bowls, lists of instructional resources and materials, and the objectives and evaluation. (CMK)

  19. Workforce Reductions: A Responsible Approach.

    ERIC Educational Resources Information Center

    National Alliance of Business, Inc., Washington, DC.

    This guide is intended to serve as a reference tool to individuals responsible for planning and implementing a work force reduction program. The information included in the guide represents a synthesis of practices that have worked for a number of companies, individuals, and communities that have had to cope with a work force reduction. The first…

  20. Noise Reduction by Signal Accumulation

    ERIC Educational Resources Information Center

    Kraftmakher, Yaakov

    2006-01-01

    The aim of this paper is to show how the noise reduction by signal accumulation can be accomplished with a data acquisition system. This topic can be used for student projects. In many cases, the noise reduction is an unavoidable part of experimentation. Several techniques are known for this purpose, and among them the signal accumulation is the…

  1. Reduction of chemical reaction models

    NASA Technical Reports Server (NTRS)

    Frenklach, Michael

    1991-01-01

    An attempt is made to reconcile the different terminologies pertaining to reduction of chemical reaction models. The approaches considered include global modeling, response modeling, detailed reduction, chemical lumping, and statistical lumping. The advantages and drawbacks of each of these methods are pointed out.

  2. Biochemistry of Catabolic Reductive Dehalogenation.

    PubMed

    Fincker, Maeva; Spormann, Alfred M

    2017-06-20

    A wide range of phylogenetically diverse microorganisms couple the reductive dehalogenation of organohalides to energy conservation. Key enzymes of such anaerobic catabolic pathways are corrinoid and Fe-S cluster-containing, membrane-associated reductive dehalogenases. These enzymes catalyze the reductive elimination of a halide and constitute the terminal reductases of a short electron transfer chain. Enzymatic and physiological studies revealed the existence of quinone-dependent and quinone-independent reductive dehalogenases that are distinguishable at the amino acid sequence level, implying different modes of energy conservation in the respective microorganisms. In this review, we summarize current knowledge about catabolic reductive dehalogenases and the electron transfer chain they are part of. We review reaction mechanisms and the role of the corrinoid and Fe-S cluster cofactors and discuss physiological implications.

  3. Technologies for Aircraft Noise Reduction

    NASA Technical Reports Server (NTRS)

    Huff, Dennis L.

    2006-01-01

    Technologies for aircraft noise reduction have been developed by NASA over the past 15 years through the Advanced Subsonic Technology (AST) Noise Reduction Program and the Quiet Aircraft Technology (QAT) project. This presentation summarizes highlights from these programs and anticipated noise reduction benefits for communities surrounding airports. Historical progress in noise reduction and technologies available for future aircraft/engine development are identified. Technologies address aircraft/engine components including fans, exhaust nozzles, landing gear, and flap systems. New "chevron" nozzles have been developed and implemented on several aircraft in production today that provide significant jet noise reduction. New engines using Ultra-High Bypass (UHB) ratios are projected to provide about 10 EPNdB (Effective Perceived Noise Level in decibels) engine noise reduction relative to the average fleet that was flying in 1997. Audio files are embedded in the presentation that estimate the sound levels for a 35,000 pound thrust engine for takeoff and approach power conditions. The predictions are based on actual model scale data that was obtained by NASA. Finally, conceptual pictures are shown that look toward future aircraft/propulsion systems that might be used to obtain further noise reduction.

  4. Bromate Reduction by Denitrifying Bacteria

    PubMed Central

    Hijnen, W.; Voogt, R.; Veenendaal, H. R.; van der Jagt, H.; van der Kooij, D.

    1995-01-01

    In the presence of bromide, ozonation as applied in water treatment results in the formation of bromate, an ion with carcinogenic properties. The reduction of bromate by mixed bacterial populations as well as pure cultures was studied under laboratory conditions. Bromate was reduced to bromide by a mixed bacterial population with and without a preceding nitrate reduction step in an anaerobically incubated medium with ethanol as the energy and carbon source at 20 and 25 deg C. The predominating bacteria isolated from the batches showing bromate reduction were identified as Pseudomonas spp. Strains of Pseudomonas fluorescens reduced BrO(inf3)(sup-) to Br(sup-) but at a much lower rate than the mixed bacterial population did. Nitrate is a preferred electron acceptor for the bromate-reducing bacteria. Bromate reduction did not occur in the presence of NO(inf3)(sup-), and the rate of bromate reduction was at least 100 times lower than the rate of nitrate reduction. Bromate was completely converted to Br(sup-), indicating that intermediates, e.g., BrO(inf2)(sup-), did not accumulate during bromate reduction. PMID:16534907

  5. Drugs, prisons, and harm reduction.

    PubMed

    Hughes, Rhidian

    2003-01-01

    The use of drugs in society raises important considerations for health and social policy. Critical health and social care issues arise when drugs are used inside prisons. This paper argues that there is an urgent need for prison drug policies to adopt the principles of harm reduction. However, current policy orthodoxy emphasises the control of drugs and punishment for drug taking. Key components of harm reduction are operationalised in this article by exploring the potential for harm reduction in prison within the context of English drug policy. Whilst the focus is on English policy debates, the discussion will have wider international resonance. Copyright 2003 The Haworth Press, Inc.

  6. Subsolidus reduction of lunar spinels.

    NASA Technical Reports Server (NTRS)

    Haggerty, S. E.

    1971-01-01

    Discussion of evidence that some lunar basalts must have exceeded the lower limit of crystallization oxygen fugacity (fO2) by several orders of magnitude. The evidence is based primarily on the decomposition of Cr-Al-ulvospinel, and is further supported in one case by the decomposition of olivine. The data suggest that some rocks have undergone intense nonequilibrium subsolidus reduction. The reduction phenomenon is widespread, and is considered to have developed either during initial deuteric cooling or as a result of a postcrystallization reduction event.

  7. Geometric Quantization and Foliation Reduction

    NASA Astrophysics Data System (ADS)

    Skerritt, Paul

    A standard question in the study of geometric quantization is whether symplectic reduction interacts nicely with the quantized theory, and in particular whether "quantization commutes with reduction." Guillemin and Sternberg first proposed this question, and answered it in the affirmative for the case of a free action of a compact Lie group on a compact Kahler manifold. Subsequent work has focused mainly on extending their proof to non-free actions and non-Kahler manifolds. For realistic physical examples, however, it is desirable to have a proof which also applies to non-compact symplectic manifolds. In this thesis we give a proof of the quantization-reduction problem for general symplectic manifolds. This is accomplished by working in a particular wavefunction representation, associated with a polarization that is in some sense compatible with reduction. While the polarized sections described by Guillemin and Sternberg are nonzero on a dense subset of the Kahler manifold, the ones considered here are distributional, having support only on regions of the phase space associated with certain quantized, or "admissible", values of momentum. We first propose a reduction procedure for the prequantum geometric structures that "covers" symplectic reduction, and demonstrate how both symplectic and prequantum reduction can be viewed as examples of foliation reduction. Consistency of prequantum reduction imposes the above-mentioned admissibility conditions on the quantized momenta, which can be seen as analogues of the Bohr-Wilson-Sommerfeld conditions for completely integrable systems. We then describe our reduction-compatible polarization, and demonstrate a one-to-one correspondence between polarized sections on the unreduced and reduced spaces. Finally, we describe a factorization of the reduced prequantum bundle, suggested by the structure of the underlying reduced symplectic manifold. This in turn induces a factorization of the space of polarized sections that agrees

  8. Fan Noise Reduction: An Overview

    NASA Technical Reports Server (NTRS)

    Envia, Edmane

    2001-01-01

    Fan noise reduction technologies developed as part of the engine noise reduction element of the Advanced Subsonic Technology Program are reviewed. Developments in low-noise fan stage design, swept and leaned outlet guide vanes, active noise control, fan flow management, and scarfed inlet are discussed. In each case, a description of the method is presented and, where available, representative results and general conclusions are discussed. The review concludes with a summary of the accomplishments of the AST-sponsored fan noise reduction research and a few thoughts on future work.

  9. Reductive Degradation: Versatile, Low Cost.

    ERIC Educational Resources Information Center

    Water and Sewage Works, 1979

    1979-01-01

    This article discusses the use of reductive degradation as an economical and effective treatment of chlorinated hydrocarbons. Comparisons with activated carbon treatment show lower capital equipment and treatment costs. (CS)

  10. Gearless speed-reduction motor

    NASA Technical Reports Server (NTRS)

    Madey, J.

    1977-01-01

    Proposed rolling electric motor has output shaft speed reductions of 1000 to 1 or better. Light compact unit uses no gears or pulleys to reduce speed presenting less bulk and frictional loss, and more efficiency.

  11. Reductive Degradation: Versatile, Low Cost.

    ERIC Educational Resources Information Center

    Water and Sewage Works, 1979

    1979-01-01

    This article discusses the use of reductive degradation as an economical and effective treatment of chlorinated hydrocarbons. Comparisons with activated carbon treatment show lower capital equipment and treatment costs. (CS)

  12. Trends in breast reduction technique.

    PubMed

    Wolf, Omer; Westreich, Melvyn; Shalom, Avshalom

    2012-05-01

    There are two main approaches to breast reduction surgery today: the traditional long scar ("Wise-pattern") technique and the more recent short ("vertical") scar technique, which is becoming more popular. During the last two decades there has been a gradual shift between the two techniques, including at our institute. To evaluate the evidence behind this obvious trend. We retrospectively collected data fromarchived hospital charts of all patients who underwent breast reduction surgery during the period 1995-2007. Epidemicological, clinical and postoperative data were analyzed and compared between patients in whom the short scar vs. the long scar techniques was used. During the study period 91 patients underwent breast reduction surgery in our department: 34 with the Wise-pattern breast reduction technique and 57 with the short-scar procedure. There was no significant difference in operative and postoperative data, including length of hospital stay. In some of the categories there was even a slight advantage (but not statistically significant) to the former. The only significant difference was the size of reduction, with a tendencyto prefer the long scar technique for larger reductions; however, with experience gained the limit for short scar reductions was gradually extended to a maximum of 1470 g. We noticed a sharp increase in the safe and uneventful practice of the short scar technique in breast reduction of < or = 1400 g--especially in young women without extreme ptosis. This observation, together with other advantages, namely, reduced scar length, prolonged shape preservation and better breast projection, support the use of this technique.

  13. Metal reduction kinetics in Shewanella.

    PubMed

    Lall, Raman; Mitchell, Julie

    2007-10-15

    Metal reduction kinetics have been studied in cultures of dissimilatory metal reducing bacteria which include the Shewanella oneidensis strain MR-1. Estimation of system parameters from time-series data faces obstructions in the implementation depending on the choice of the mathematical model that captures the observed dynamics. The modeling of metal reduction is often based on Michaelis-Menten equations. These models are often developed using initial in vitro reaction rates and seldom match with in vivo reduction profiles. For metal reduction studies, we propose a model that is based on the power law representation that is effectively applied to the kinetics of metal reduction. The method yields reasonable parameter estimates and is illustrated with the analysis of time-series data that describes the dynamics of metal reduction in S.oneidensis strain MR-1. In addition, mixed metal studies involving the reduction of Uranyl (U(VI)) to the relatively insoluble tetravalent form (U(IV)) by S. alga strain (BR-Y) were studied in the presence of environmentally relevant iron hydrous oxides. For mixed metals, parameter estimation and curve fitting are accomplished with a generalized least squares formulation that handles systems of ordinary differential equations and is implemented in Matlab. It consists of an optimization algorithm (Levenberg-Marquardt, LSQCURVEFIT) and a numerical ODE solver. Simulation with the estimated parameters indicates that the model captures the experimental data quite well. The model uses the estimated parameters to predict the reduction rates of metals and mixed metals at varying concentrations. Supplementary data are available at Bioinformatics online.

  14. 2dfdr: Data reduction software

    NASA Astrophysics Data System (ADS)

    AAO software Team

    2015-05-01

    2dfdr is an automatic data reduction pipeline dedicated to reducing multi-fibre spectroscopy data, with current implementations for AAOmega (fed by the 2dF, KOALA-IFU, SAMI Multi-IFU or older SPIRAL front-ends), HERMES, 2dF (spectrograph), 6dF, and FMOS. A graphical user interface is provided to control data reduction and allow inspection of the reduced spectra.

  15. Different phycobilin antenna organisations affect the balance between light use and growth rate in the cyanobacterium Microcystis aeruginosa and in the cryptophyte Cryptomonas ovata.

    PubMed

    Kunath, Christfried; Jakob, Torsten; Wilhelm, Christian

    2012-03-01

    During the recent years, wide varieties of methodologies have been developed up to the level of commercial use to measure photosynthetic electron transport by modulated chlorophyll a-in vivo fluorescence. It is now widely accepted that the ratio between electron transport rates and new biomass (P (Fl)/B (C)) is not fixed and depends on many factors that are also taxonomically variable. In this study, the balance between photon absorption and biomass production has been measured in two phycobilin-containing phototrophs, namely, a cyanobacterium and a cryptophyte, which differ in their antenna organization. It is demonstrated that the different antenna organization exerts influence on the regulation of the primary photosynthetic reaction and the dissipation of excessively absorbed radiation. Although, growth rates and the quantum efficiency of biomass production of both phototrophs were comparable, the ratio P (Fl)/B (C) was twice as high in the cryptophyte in comparison to the cyanobacterium. It is assumed that this discrepancy is because of differences in the metabolic regulation of cell growth. In the cryptophyte, absorbed photosynthetic energy is used to convert assimilated carbon directly into proteins and lipids, whereas in the cyanobacterium, the photosynthetic energy is preferentially stored as carbohydrates.

  16. [Substances considered addictive: prohibition, harm reduction and risk reduction].

    PubMed

    Menéndez, Eduardo

    2012-01-01

    Latin America is currently the region with the highest rate of homicides worldwide, and a large part of the killings are linked to so-called organized crime, especially drug trafficking. The trafficking of drugs is a consequence of the illegality of certain substances which - at least presently - is based in and legitimated by biomedical criteria that turns the production, commercialization and often the consumption of certain substances considered addictive into "offenses against health." This text briefly analyzes the two policies formulated and implemented thus far in terms of prohibition and harm reduction, considering the failure of prohibitionism as well as the limitations of harm reduction proposals. The constant and multiple inconsistencies and contradictions of prohibitionism are noted, indicating the necessity of regarding cautiously repeated comments about its "failure." The text proposes the implementation of a policy of risk reduction that includes not only the behavior of individuals and groups, but also the structural dimension, both in economic-political and cultural terms.

  17. Evolutionary Dynamics of Cryptophyte Plastid Genomes

    PubMed Central

    Kim, Jong Im; Moore, Christa E.; Archibald, John M.; Bhattacharya, Debashish; Yi, Gangman

    2017-01-01

    Abstract Cryptophytes are an ecologically important group of largely photosynthetic unicellular eukaryotes. This lineage is of great interest to evolutionary biologists because their plastids are of red algal secondary endosymbiotic origin and the host cell retains four different genomes (host nuclear, mitochondrial, plastid, and red algal nucleomorph). Here, we report a comparative analysis of plastid genomes from six representative cryptophyte genera. Four newly sequenced cryptophyte plastid genomes of Chroomonas mesostigmatica, Ch. placoidea, Cryptomonas curvata, and Storeatula sp. CCMP1868 share a number of features including synteny and gene content with the previously sequenced genomes of Cryptomonas paramecium, Rhodomonas salina, Teleaulax amphioxeia, and Guillardia theta. Our analysis of these plastid genomes reveals examples of gene loss and intron insertion. In particular, the chlB/chlL/chlN genes, which encode light-independent (dark active) protochlorophyllide oxidoreductase (LIPOR) proteins have undergone recent gene loss and pseudogenization in cryptophytes. Comparison of phylogenetic trees based on plastid and nuclear genome data sets show the introduction, via secondary endosymbiosis, of a red algal derived plastid in a lineage of chlorophyll-c containing algae. This event was followed by additional rounds of eukaryotic endosymbioses that spread the red lineage plastid to diverse groups such as haptophytes and stramenopiles. PMID:28854597

  18. A Virtual Aluminum Reduction Cell

    NASA Astrophysics Data System (ADS)

    Zhang, Hongliang; Zhou, Chenn Q.; Wu, Bing; Li, Jie

    2013-11-01

    The most important component in the aluminum industry is the aluminum reduction cell; it has received considerable interests and resources to conduct research to improve its productivity and energy efficiency. The current study focused on the integration of numerical simulation data and virtual reality technology to create a scientifically and practically realistic virtual aluminum reduction cell by presenting complex cell structures and physical-chemical phenomena. The multiphysical field simulation models were first built and solved in ANSYS software (ANSYS Inc., Canonsburg, PA, USA). Then, the methodology of combining the simulation results with virtual reality was introduced, and a virtual aluminum reduction cell was created. The demonstration showed that a computer-based world could be created in which people who are not analysis experts can see the detailed cell structure in a context that they can understand easily. With the application of the virtual aluminum reduction cell, even people who are familiar with aluminum reduction cell operations can gain insights that make it possible to understand the root causes of observed problems and plan design changes in much less time.

  19. Midwestern Greenhouse Gas Reduction Accord

    SciTech Connect

    2007-07-01

    The Midwestern Greenhouse Gas Reduction Acccord, or Midwestern Greenhouse gas Accord (MGA), is a regional agreement by governors of the states in the US Midwest and one Canadian province to reduce greenhouse gas emissions to combat climate change. Signatories to the accord include the US states of Minnesota, Wisconsin, Illinois, Indiana, Iowa, Michigan, Kansas, Ohio and South Dakota, and the Canadian Province of Manitoba. The accord, signed on November 15, 2007, established the Midwestern Greenhouse Gas Reduction Program, which aims to: establish greenhouse gas reduction targets and timeframes consistent with MGA member states' targets; develop a market-based and multi-sector cap-and-trade mechanism to help achieve those reduction targets; establish a system to enable tracking, management, and crediting for entities that reduce greenhouse gas emissions; and develop and implement additional steps as needed to achieve the reduction targets, such as a low-carbon fuel standards and regional incentives and funding mechanisms. The GHG registry will be managed by the Climate Registry, which manages the registry for other US state schemes. One of the first actions was to convene an Energy Security under Climate Stewardship Platform to guide future development of the Midwest's energy economy.

  20. Manual reduction in acute haemorrhoids.

    PubMed

    Gaj, F; Candeloro, L; Biviano, I

    2016-01-01

    In prolapsed internal hemorrhoids exposed outside the anus, manually reducing the prolapse with 48 hours of commencement of anal pain, decreased the progression of thrombosis. The aim of our study was to evaluate the effects of manual reduction of the inflamed piles hemorrhoids. Eleven patients, 7 males and 4 pregnant females (in early post partum) with an average age of 34 years ± 8 (range 23- 52) were enrolled with anal pain cause by haemorrhoidal congestion, but prior to full blown thrombosis. Patients underwent a manual reduction of the external prolapsed haemorrhoidal plexus. In the 48 hours following the procedure, patients were instructed on how to insert any prolapsed hemorrhoid (piles) themselves. Pain intensity was measured using the visual analog scale (VAS) at time of consultation and then 10 days after the reduction. At day 10 following treatment we observed a statistically significant reduction in anal swelling (11 vs 1, n° pzt; p = 0.001), anal pain (11 vs 2, n° pzt; p = 0.001) and VAS score (8.6 ± 0.7 vs 0.4 ± 1.2; p = 0.001). Two patients (18%) underwent surgical haemorrhoidectomy sec. Milligan Morgan and 1 patient (9%) underwent excision of thrombosed external hemorrhoids. 73% of patients did not require surgery. Manual reduction of the prolapsed piles outside the anus decreased pain immediately and it also allows postponement of surgery or any other treatment.

  1. Microbial reduction of iron ore

    DOEpatents

    Hoffmann, Michael R.; Arnold, Robert G.; Stephanopoulos, Gregory

    1989-01-01

    A process is provided for reducing iron ore by treatment with microorganisms which comprises forming an aqueous mixture of iron ore, microorganisms operable for reducing the ferric iron of the iron ore to ferrous iron, and a substrate operable as an energy source for the microbial reduction; and maintaining the aqueous mixture for a period of time and under conditions operable to effect the reduction of the ore. Preferably the microorganism is Pseudomonas sp. 200 and the reduction conducted anaerobically with a domestic wastewater as the substrate. An aqueous solution containing soluble ferrous iron can be separated from the reacted mixture, treated with a base to precipitate ferrous hydroxide which can then be recovered as a concentrated slurry.

  2. Microbial reduction of iron ore

    DOEpatents

    Hoffmann, M.R.; Arnold, R.G.; Stephanopoulos, G.

    1989-11-14

    A process is provided for reducing iron ore by treatment with microorganisms which comprises forming an aqueous mixture of iron ore, microorganisms operable for reducing the ferric iron of the iron ore to ferrous iron, and a substrate operable as an energy source for the microbial reduction; and maintaining the aqueous mixture for a period of time and under conditions operable to effect the reduction of the ore. Preferably the microorganism is Pseudomonas sp. 200 and the reduction conducted anaerobically with a domestic wastewater as the substrate. An aqueous solution containing soluble ferrous iron can be separated from the reacted mixture, treated with a base to precipitate ferrous hydroxide which can then be recovered as a concentrated slurry. 11 figs.

  3. Noise reduction for vocal pathologies.

    PubMed

    Matassini, L; Manfredi, C

    2002-01-01

    A noise reduction scheme, particularly suited for the correction of vocal pathologies, is proposed. The filter makes use of concepts originated within the theory of dynamical systems and deterministic chaos. In particular, the idea of embedding scalar data in order to reconstruct a phase space is of fundamental importance here. Furthermore, the concept of an attractor as a result of dynamical constraints is exploited. In order to perform noise reduction one needs redundancy and the human voice provides it even within a phoneme, namely the smallest structural unit of speech. Due to several repetitions of a pattern called pitch inside a phoneme, separation between the pure voice signal and the noise is possible, provided the latter is uncorrelated with the former. With a proper parameter tuning, different kinds of noise can be removed. We describe the idea behind the noise reduction algorithm and present applications to vocal pathologies.

  4. Thermodynamics of lunar ilmenite reduction

    NASA Technical Reports Server (NTRS)

    Altenberg, B. H.; Franklin, H. A.; Jones, C. H.

    1993-01-01

    With the prospect of returning to the moon, the development of a lunar occupation would fulfill one of the goals of the Space Exploration Initiative (SEI) of the late 1980's. Processing lunar resources into useful products, such as liquid oxygen for fuel and life support, would be one of many aspects of an active lunar base. ilmenite (FeTiO3) is found on the lunar surface and can be used as a feed stock to produce oxygen. Understanding the various ilmenite-reduction reactions elucidates many processing options. Defining the thermodynamic chemical behavior at equilibrium under various conditions of temperature and pressures can be helpful in specifying optimal operating conditions. Differences between a previous theoretical analysis and experimentally determined results has sparked interest in trying to understand the effect of operating pressure on the hydrogen-reduction-of-ilmenite reaction. Various aspects of this reduction reaction are discussed.

  5. Thermodynamics of lunar ilmenite reduction

    NASA Technical Reports Server (NTRS)

    Altenberg, B. H.; Franklin, H. A.; Jones, C. H.

    1993-01-01

    With the prospect of returning to the moon, the development of a lunar occupation would fulfill one of the goals of the Space Exploration Initiative (SEI) of the late 1980's. Processing lunar resources into useful products, such as liquid oxygen for fuel and life support, would be one of many aspects of an active lunar base. ilmenite (FeTiO3) is found on the lunar surface and can be used as a feed stock to produce oxygen. Understanding the various ilmenite-reduction reactions elucidates many processing options. Defining the thermodynamic chemical behavior at equilibrium under various conditions of temperature and pressures can be helpful in specifying optimal operating conditions. Differences between a previous theoretical analysis and experimentally determined results has sparked interest in trying to understand the effect of operating pressure on the hydrogen-reduction-of-ilmenite reaction. Various aspects of this reduction reaction are discussed.

  6. Uranium isotopes fingerprint biotic reduction

    SciTech Connect

    Stylo, Malgorzata; Neubert, Nadja; Wang, Yuheng; Monga, Nikhil; Romaniello, Stephen J.; Weyer, Stefan; Bernier-Latmani, Rizlan

    2015-04-20

    Knowledge of paleo-redox conditions in the Earth’s history provides a window into events that shaped the evolution of life on our planet. The role of microbial activity in paleo-redox processes remains unexplored due to the inability to discriminate biotic from abiotic redox transformations in the rock record. The ability to deconvolute these two processes would provide a means to identify environmental niches in which microbial activity was prevalent at a specific time in paleo-history and to correlate specific biogeochemical events with the corresponding microbial metabolism. Here, we demonstrate that the isotopic signature associated with microbial reduction of hexavalent uranium (U), i.e., the accumulation of the heavy isotope in the U(IV) phase, is readily distinguishable from that generated by abiotic uranium reduction in laboratory experiments. Thus, isotope signatures preserved in the geologic record through the reductive precipitation of uranium may provide the sought-after tool to probe for biotic processes. Because uranium is a common element in the Earth’s crust and a wide variety of metabolic groups of microorganisms catalyze the biological reduction of U(VI), this tool is applicable to a multiplicity of geological epochs and terrestrial environments. The findings of this study indicate that biological activity contributed to the formation of many authigenic U deposits, including sandstone U deposits of various ages, as well as modern, Cretaceous, and Archean black shales. In addition, engineered bioremediation activities also exhibit a biotic signature, suggesting that, although multiple pathways may be involved in the reduction, direct enzymatic reduction contributes substantially to the immobilization of uranium.

  7. Uranium isotopes fingerprint biotic reduction

    DOE PAGES

    Stylo, Malgorzata; Neubert, Nadja; Wang, Yuheng; ...

    2015-04-20

    Knowledge of paleo-redox conditions in the Earth’s history provides a window into events that shaped the evolution of life on our planet. The role of microbial activity in paleo-redox processes remains unexplored due to the inability to discriminate biotic from abiotic redox transformations in the rock record. The ability to deconvolute these two processes would provide a means to identify environmental niches in which microbial activity was prevalent at a specific time in paleo-history and to correlate specific biogeochemical events with the corresponding microbial metabolism. Here, we demonstrate that the isotopic signature associated with microbial reduction of hexavalent uranium (U),more » i.e., the accumulation of the heavy isotope in the U(IV) phase, is readily distinguishable from that generated by abiotic uranium reduction in laboratory experiments. Thus, isotope signatures preserved in the geologic record through the reductive precipitation of uranium may provide the sought-after tool to probe for biotic processes. Because uranium is a common element in the Earth’s crust and a wide variety of metabolic groups of microorganisms catalyze the biological reduction of U(VI), this tool is applicable to a multiplicity of geological epochs and terrestrial environments. The findings of this study indicate that biological activity contributed to the formation of many authigenic U deposits, including sandstone U deposits of various ages, as well as modern, Cretaceous, and Archean black shales. In addition, engineered bioremediation activities also exhibit a biotic signature, suggesting that, although multiple pathways may be involved in the reduction, direct enzymatic reduction contributes substantially to the immobilization of uranium.« less

  8. Technologies for Turbofan Noise Reduction

    NASA Technical Reports Server (NTRS)

    Huff, Dennis

    2005-01-01

    An overview presentation of NASA's engine noise research since 1992 is given for subsonic commercial aircraft applications. Highlights are included from the Advanced Subsonic Technology (AST) Noise Reduction Program and the Quiet Aircraft Technology (QAT) project with emphasis on engine source noise reduction. Noise reduction goals for 10 EPNdB by 207 and 20 EPNdB by 2022 are reviewed. Fan and jet noise technologies are highlighted from the AST program including higher bypass ratio propulsion, scarf inlets, forward-swept fans, swept/leaned stators, chevron nozzles, noise prediction methods, and active noise control for fans. Source diagnostic tests for fans and jets that have been completed over the past few years are presented showing how new flow measurement methods such as Particle Image Velocimetry (PIV) have played a key role in understanding turbulence, the noise generation process, and how to improve noise prediction methods. Tests focused on source decomposition have helped identify which engine components need further noise reduction. The role of Computational AeroAcoustics (CAA) for fan noise prediction is presented. Advanced noise reduction methods such as Hershel-Quincke tubes and trailing edge blowing for fan noise that are currently being pursued n the QAT program are also presented. Highlights are shown form engine validation and flight demonstrations that were done in the late 1990's with Pratt & Whitney on their PW4098 engine and Honeywell on their TFE-731-60 engine. Finally, future propulsion configurations currently being studied that show promise towards meeting NASA's long term goal of 20 dB noise reduction are shown including a Dual Fan Engine concept on a Blended Wing Body aircraft.

  9. Uranium isotopes fingerprint biotic reduction

    PubMed Central

    Stylo, Malgorzata; Neubert, Nadja; Wang, Yuheng; Monga, Nikhil; Romaniello, Stephen J.; Weyer, Stefan; Bernier-Latmani, Rizlan

    2015-01-01

    Knowledge of paleo-redox conditions in the Earth’s history provides a window into events that shaped the evolution of life on our planet. The role of microbial activity in paleo-redox processes remains unexplored due to the inability to discriminate biotic from abiotic redox transformations in the rock record. The ability to deconvolute these two processes would provide a means to identify environmental niches in which microbial activity was prevalent at a specific time in paleo-history and to correlate specific biogeochemical events with the corresponding microbial metabolism. Here, we demonstrate that the isotopic signature associated with microbial reduction of hexavalent uranium (U), i.e., the accumulation of the heavy isotope in the U(IV) phase, is readily distinguishable from that generated by abiotic uranium reduction in laboratory experiments. Thus, isotope signatures preserved in the geologic record through the reductive precipitation of uranium may provide the sought-after tool to probe for biotic processes. Because uranium is a common element in the Earth’s crust and a wide variety of metabolic groups of microorganisms catalyze the biological reduction of U(VI), this tool is applicable to a multiplicity of geological epochs and terrestrial environments. The findings of this study indicate that biological activity contributed to the formation of many authigenic U deposits, including sandstone U deposits of various ages, as well as modern, Cretaceous, and Archean black shales. Additionally, engineered bioremediation activities also exhibit a biotic signature, suggesting that, although multiple pathways may be involved in the reduction, direct enzymatic reduction contributes substantially to the immobilization of uranium. PMID:25902522

  10. Uranium isotopes fingerprint biotic reduction.

    PubMed

    Stylo, Malgorzata; Neubert, Nadja; Wang, Yuheng; Monga, Nikhil; Romaniello, Stephen J; Weyer, Stefan; Bernier-Latmani, Rizlan

    2015-05-05

    Knowledge of paleo-redox conditions in the Earth's history provides a window into events that shaped the evolution of life on our planet. The role of microbial activity in paleo-redox processes remains unexplored due to the inability to discriminate biotic from abiotic redox transformations in the rock record. The ability to deconvolute these two processes would provide a means to identify environmental niches in which microbial activity was prevalent at a specific time in paleo-history and to correlate specific biogeochemical events with the corresponding microbial metabolism. Here, we demonstrate that the isotopic signature associated with microbial reduction of hexavalent uranium (U), i.e., the accumulation of the heavy isotope in the U(IV) phase, is readily distinguishable from that generated by abiotic uranium reduction in laboratory experiments. Thus, isotope signatures preserved in the geologic record through the reductive precipitation of uranium may provide the sought-after tool to probe for biotic processes. Because uranium is a common element in the Earth's crust and a wide variety of metabolic groups of microorganisms catalyze the biological reduction of U(VI), this tool is applicable to a multiplicity of geological epochs and terrestrial environments. The findings of this study indicate that biological activity contributed to the formation of many authigenic U deposits, including sandstone U deposits of various ages, as well as modern, Cretaceous, and Archean black shales. Additionally, engineered bioremediation activities also exhibit a biotic signature, suggesting that, although multiple pathways may be involved in the reduction, direct enzymatic reduction contributes substantially to the immobilization of uranium.

  11. 2008 world direct reduction statistics

    SciTech Connect

    2009-07-01

    This supplement discusses total direct reduced iron (DRI) production for 2007 and 2008 by process. Total 2008 production by MIDREX(reg sign) direct reduction process plants was over 39.8 million tons. The total of all coal-based processes was 17.6 million tons. Statistics for world DRI production are also given by region for 2007 and 2008 and by year (1970-2009). Capacity utilization for 2008 by process is given. World DRI production by region and by process is given for 1998-2008 and world DRI shipments are given from the 1970s to 2008. A list of world direct reduction plants is included.

  12. Magnesium reduction of uranium oxide

    SciTech Connect

    Elliott, G.R.B.

    1985-08-13

    A method and apparatus are provided for reducing uranium oxide with magnesium to form uranium metal. The reduction is carried out in a molten-salt solution of density greater than 3.4 grams per cubic centimeter, thereby allowing the uranium product to sink and the magnesium oxide byproduct to float, consequently allowing separation of product and byproduct.

  13. Background reduction in cryogenic detectors

    SciTech Connect

    Bauer, Daniel A.; /Fermilab

    2005-04-01

    This paper discusses the background reduction and rejection strategy of the Cryogenic Dark Matter Search (CDMS) experiment. Recent measurements of background levels from CDMS II at Soudan are presented, along with estimates for future improvements in sensitivity expected for a proposed SuperCDMS experiment at SNOLAB.

  14. Sex-work harm reduction.

    PubMed

    Rekart, Michael L

    2005-12-17

    Sex work is an extremely dangerous profession. The use of harm-reduction principles can help to safeguard sex workers' lives in the same way that drug users have benefited from drug-use harm reduction. Sex workers are exposed to serious harms: drug use, disease, violence, discrimination, debt, criminalisation, and exploitation (child prostitution, trafficking for sex work, and exploitation of migrants). Successful and promising harm-reduction strategies are available: education, empowerment, prevention, care, occupational health and safety, decriminalisation of sex workers, and human-rights-based approaches. Successful interventions include peer education, training in condom-negotiating skills, safety tips for street-based sex workers, male and female condoms, the prevention-care synergy, occupational health and safety guidelines for brothels, self-help organisations, and community-based child protection networks. Straightforward and achievable steps are available to improve the day-to-day lives of sex workers while they continue to work. Conceptualising and debating sex-work harm reduction as a new paradigm can hasten this process.

  15. Workforce Reductions. An Annotated Bibliography.

    ERIC Educational Resources Information Center

    Hickok, Thomas A.; Hickok, Thomas A.

    This report, which is based on a review of practitioner-oriented sources and scholarly journals, uses a three-part framework to organize annotated bibliographies that, together, list a total of 104 sources that provide the following three perspectives on work force reduction issues: organizational, organizational-individual relationship, and…

  16. Graded geometry and Poisson reduction

    SciTech Connect

    Cattaneo, A. S.; Zambon, M.

    2009-02-02

    The main result extends the Marsden-Ratiu reduction theorem in Poisson geometry, and is proven by means of graded geometry. In this note we provide the background material about graded geometry necessary for the proof. Further, we provide an alternative algebraic proof for the main result.

  17. Dimensional Reduction and Hadronic Processes

    SciTech Connect

    Signer, Adrian; Stoeckinger, Dominik

    2008-11-23

    We consider the application of regularization by dimensional reduction to NLO corrections of hadronic processes. The general collinear singularity structure is discussed, the origin of the regularization-scheme dependence is identified and transition rules to other regularization schemes are derived.

  18. Flavin reduction activates Drosophila cryptochrome.

    PubMed

    Vaidya, Anand T; Top, Deniz; Manahan, Craig C; Tokuda, Joshua M; Zhang, Sheng; Pollack, Lois; Young, Michael W; Crane, Brian R

    2013-12-17

    Entrainment of circadian rhythms in higher organisms relies on light-sensing proteins that communicate to cellular oscillators composed of delayed transcriptional feedback loops. The principal photoreceptor of the fly circadian clock, Drosophila cryptochrome (dCRY), contains a C-terminal tail (CTT) helix that binds beside a FAD cofactor and is essential for light signaling. Light reduces the dCRY FAD to an anionic semiquinone (ASQ) radical and increases CTT proteolytic susceptibility but does not lead to CTT chemical modification. Additional changes in proteolytic sensitivity and small-angle X-ray scattering define a conformational response of the protein to light that centers at the CTT but also involves regions remote from the flavin center. Reduction of the flavin is kinetically coupled to CTT rearrangement. Chemical reduction to either the ASQ or the fully reduced hydroquinone state produces the same conformational response as does light. The oscillator protein Timeless (TIM) contains a sequence similar to the CTT; the corresponding peptide binds dCRY in light and protects the flavin from oxidation. However, TIM mutants therein still undergo dCRY-mediated degradation. Thus, photoreduction to the ASQ releases the dCRY CTT and promotes binding to at least one region of TIM. Flavin reduction by either light or cellular reductants may be a general mechanism of CRY activation.

  19. Flavin reduction activates Drosophila cryptochrome

    PubMed Central

    Vaidya, Anand T.; Top, Deniz; Manahan, Craig C.; Tokuda, Joshua M.; Zhang, Sheng; Pollack, Lois; Young, Michael W.; Crane, Brian R.

    2013-01-01

    Entrainment of circadian rhythms in higher organisms relies on light-sensing proteins that communicate to cellular oscillators composed of delayed transcriptional feedback loops. The principal photoreceptor of the fly circadian clock, Drosophila cryptochrome (dCRY), contains a C-terminal tail (CTT) helix that binds beside a FAD cofactor and is essential for light signaling. Light reduces the dCRY FAD to an anionic semiquinone (ASQ) radical and increases CTT proteolytic susceptibility but does not lead to CTT chemical modification. Additional changes in proteolytic sensitivity and small-angle X-ray scattering define a conformational response of the protein to light that centers at the CTT but also involves regions remote from the flavin center. Reduction of the flavin is kinetically coupled to CTT rearrangement. Chemical reduction to either the ASQ or the fully reduced hydroquinone state produces the same conformational response as does light. The oscillator protein Timeless (TIM) contains a sequence similar to the CTT; the corresponding peptide binds dCRY in light and protects the flavin from oxidation. However, TIM mutants therein still undergo dCRY-mediated degradation. Thus, photoreduction to the ASQ releases the dCRY CTT and promotes binding to at least one region of TIM. Flavin reduction by either light or cellular reductants may be a general mechanism of CRY activation. PMID:24297896

  20. Palladium Catalyzed Reduction of Nitrobenzene.

    ERIC Educational Resources Information Center

    Mangravite, John A.

    1983-01-01

    Compares two palladium (Pd/C) reducing systems to iron/tin-hydrochloric acid (Fe/HCl and Sn/HCl) reductions and suggests an efficient, clean, and inexpensive procedures for the conversion of nitrobenzene to aniline. Includes laboratory procedures used and discussion of typical results obtained. (JN)

  1. Workforce Reductions. An Annotated Bibliography.

    ERIC Educational Resources Information Center

    Hickok, Thomas A.; Hickok, Thomas A.

    This report, which is based on a review of practitioner-oriented sources and scholarly journals, uses a three-part framework to organize annotated bibliographies that, together, list a total of 104 sources that provide the following three perspectives on work force reduction issues: organizational, organizational-individual relationship, and…

  2. GumTree: Data reduction

    NASA Astrophysics Data System (ADS)

    Rayner, Hugh; Hathaway, Paul; Hauser, Nick; Fei, Yang; Franceschini, Ferdi; Lam, Tony

    2006-11-01

    Access to software tools for interactive data reduction, visualisation and analysis during a neutron scattering experiment enables instrument users to make informed decisions regarding the direction and success of their experiment. ANSTO aims to enhance the experiment experience of its facility's users by integrating these data reduction tools with the instrument control interface for immediate feedback. GumTree is a software framework and application designed to support an Integrated Scientific Experimental Environment, for concurrent access to instrument control, data acquisition, visualisation and analysis software. The Data Reduction and Analysis (DRA) module is a component of the GumTree framework that allows users to perform data reduction, correction and basic analysis within GumTree while an experiment is running. It is highly integrated with GumTree, able to pull experiment data and metadata directly from the instrument control and data acquisition components. The DRA itself uses components common to all instruments at the facility, providing a consistent interface. It features familiar ISAW-based 1D and 2D plotting, an OpenGL-based 3D plotter and peak fitting performed by fityk. This paper covers the benefits of integration, the flexibility of the DRA module, ease of use for the interface and audit trail generation.

  3. A hartmann test reduction program.

    PubMed

    Schulte, D H

    1968-01-01

    A generalized Fortran program for the reduction of Hartmann test data has been written. A brief review of the mathematical echnique is given, along with a discussion of the measuring methods and the results of some tests of the accuracy of the program.

  4. Harm reduction in developing countries.

    PubMed

    Wong, C

    1998-06-29

    Manipur, a state in northeast India and one of its poorest areas, has widespread iv drug use and accompanying risky behavior. By the early 1990s, 50% of iv drug users (IDUs) there were infected with HIV. Initial efforts to implement harm reduction measures by the state government and nongovernmental organizations (NGOs) were unsuccessful because of concerns regarding community resistance. According to Khomdon Singh Lisam, Project Director of the Manipur State AIDS Control Society, it was necessary to lay the groundwork for harm reduction by involving community and professional groups; this led to the adoption of the Manipur State AIDS Policy (MSAP) by the State Cabinet. This policy permits the implementation of a number of services, including needle exchange and education about sterilizing drug equipment. According to Lisam, there is strong community support for the harm reduction program. In the city of Madras, in southern India, needle exchange is illegal (the MSAP is one of a kind in India), but needles can be bought for a few centimes at pharmacies. However, according to M. Suresh Kumar, of the Society for Aid and Help for Addictive Illnesses (SAHAI) in Madras, IDUs cannot afford money for the needles. SAHAI has been working together with influential people to develop outreach programs. Education of IDUs must be face-to-face, because illiteracy is so high; the social networks and families of IDUs are also educated about risk reduction.

  5. APPLICATION OF RADON REDUCTION METHODS

    EPA Science Inventory

    The document is intended to aid homeowners and contractors in diagnosing and solving indoor radon problems. It will also be useful to State and Federal regulatory officials and many other persons who provide advice on the selection, design and operation of radon reduction methods...

  6. Functionalization of Organotrifluoroborates: Reductive Amination

    PubMed Central

    Cooper, David J.

    2010-01-01

    Herein we report the conversion of aldehyde-containing potassium and tetrabutylammonium organotrifluoroborates to the corresponding amines through reductive amination protocols. Potassium formate facilitated by catalytic palladium acetate, sodium triacetoxyborohydride, and pyridine borane have all served as effective hydride donors, reducing the initially formed imines or iminium ions to provide the corresponding amines. PMID:18412389

  7. Palladium Catalyzed Reduction of Nitrobenzene.

    ERIC Educational Resources Information Center

    Mangravite, John A.

    1983-01-01

    Compares two palladium (Pd/C) reducing systems to iron/tin-hydrochloric acid (Fe/HCl and Sn/HCl) reductions and suggests an efficient, clean, and inexpensive procedures for the conversion of nitrobenzene to aniline. Includes laboratory procedures used and discussion of typical results obtained. (JN)

  8. HEAT ISLAND REDUCTION STRATEGIES GUIDEBOOK ...

    EPA Pesticide Factsheets

    This heat island reduction strategies guidebook provides an overview of urban heat islands and steps communities can take to reduce them. In particular, this guidebook provides background basics and answers the questions: “What is a heat island?” “What are its impacts?

  9. Reduction of postoperative adhesion development.

    PubMed

    Diamond, Michael P

    2016-10-01

    Despite use of meticulous surgical techniques, and regardless of surgical access via laparotomy or laparoscopy, postoperative adhesions develop in the vast majority of women undergoing abdominopelvic surgery. Such adhesions represent not only adhesion reformation at sites of adhesiolysis, but also de novo adhesion formation at sites of surgical procedures. Application of antiadhesion adjuvants compliment the benefits of meticulous surgical techniques, providing an opportunity to further reduce postoperative adhesion development. Improved understanding of the pathophysiology of adhesion development and distinguishing variations in the molecular biologic mechanisms from adhesion-free peritoneal repair represent future opportunities to improve the reduction of postoperative adhesions. Optimization of the reduction of postoperative adhesions will likely require identification of unique, personalized approaches in each individual, representing interindividual variation in peritoneal repair processes. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  10. Optical Waveguide Scattering Reduction. II.

    DTIC Science & Technology

    1980-12-01

    FAD-AOAR 815 BATTELLEWCOLUMBUS LABS ON F/S 20/6 OPTICAL WAVEGUIDE SCATTER ING REDUC TION. II.(U) 7 DEC 80 0 W VAHEY, N F HARTMAN, R C SHERMAN F3361... OPTICAL WAVEGUIDE SCATTERING REDUCTION II M BATTELLE COLUMBUS LABORATORIES 505 KING AVENUE COLUMBUS, OHIO 43201 DTIC ELECTEf MAY 12 198111 December...reviewed and is approved for publication. DOUGLAS AWIWILLE, Project Engineer KENNETH R. HUTCHINSON, Chief Electro- Optics Techniques and Electro- Optics

  11. Dimension Reduction for Object Ranking

    NASA Astrophysics Data System (ADS)

    Kamishima, Toshihiro; Akaho, Shotaro

    Ordered lists of objects are widely used as representational forms. Such ordered objects include Web search results and bestseller lists. Techniques for processing such ordinal data are being developed, particularly methods for an object ranking task: i.e., learning functions used to sort objects from sample orders. In this article, we propose two dimension reduction methods specifically designed to improve prediction performance in an object ranking task.

  12. Classification, Clustering and Dimensionality Reduction

    DTIC Science & Technology

    2008-07-08

    and scale remains unsolved. New and emerging applications, such as data mining , web searching, retrieval of multimedia data, face recognition and...algorithms, and (4) dimensionality reduction. Solution to these problems will advance the state-of-the-art in pattern recognition, data mining and machine...learning. These advances will also be useful to a number of pattern recognition and data mining applications of interest to the Navy.

  13. Budget Reduction in the Navy

    DTIC Science & Technology

    1990-12-01

    ORGANIZATION Naval Postgraduate School (If applicable ) Naval Postgraduate School 36 6c ADDRESS (City, State, and ZIP Code) 7b ADDRESS (City, State, and ZIP Code...ORGANIZATION (If applicable ) 8c ADDRESS (City, State, and ZIP Code) 10 SOURCE OF FUNDING NUMBERS Program fiement N. Project N. Task Nu Wor, unlt ACCewon...program reductions made by the Navy for fiscal years 1990, 1991 and beyound. Current and historical budget data were obtained from the Office of the

  14. Grounds Management Cost Reduction Strategies

    DTIC Science & Technology

    1993-06-01

    provided to flower beds. A-40 Use Plant Grouth Regulators (PGRs) None provided on semi-improved grounds. A-38-40 AIR FORCE GROUNDS NINTEIANCE COST REDUCTION...eral checklist with suggestions for renovation of grasslands. 149. A very economical way to control undesirable understory vegetation 4 in woods or to...34 (Environmental Laboratory 1986). This dou- cides, permitting economical weed control. Once ument specifically deals with the planning, layout, established

  15. Americium recovery from reduction residues

    DOEpatents

    Conner, W.V.; Proctor, S.G.

    1973-12-25

    A process for separation and recovery of americium values from container or bomb'' reduction residues comprising dissolving the residues in a suitable acid, adjusting the hydrogen ion concentration to a desired level by adding a base, precipitating the americium as americium oxalate by adding oxalic acid, digesting the solution, separating the precipitate, and thereafter calcining the americium oxalate precipitate to form americium oxide. (Official Gazette)

  16. Snowmelt Increase Through Albedo Reduction

    DTIC Science & Technology

    1988-12-01

    Studies of Snow and Ice in Hyvarinen, T. and J. Lammasnieme (1987) Mountain Regions, International Association of Infrared measurement of free-water...snow-climate feedback, and the reduction in albedo by darkening agents has been studied and practiced extensively. Although much is known about albedo...sometimes CHARACTERISTICS gets in the way of man’s activities and must be removed as quickly as possible. When snow is Many studies of crystal growth in snow

  17. Model Reduction by Manifold Boundaries

    NASA Astrophysics Data System (ADS)

    Transtrum, Mark K.; Qiu, Peng

    2014-08-01

    Understanding the collective behavior of complex systems from their basic components is a difficult yet fundamental problem in science. Existing model reduction techniques are either applicable under limited circumstances or produce "black boxes" disconnected from the microscopic physics. We propose a new approach by translating the model reduction problem for an arbitrary statistical model into a geometric problem of constructing a low-dimensional, submanifold approximation to a high-dimensional manifold. When models are overly complex, we use the observation that the model manifold is bounded with a hierarchy of widths and propose using the boundaries as submanifold approximations. We refer to this approach as the manifold boundary approximation method. We apply this method to several models, including a sum of exponentials, a dynamical systems model of protein signaling, and a generalized Ising model. By focusing on parameters rather than physical degrees of freedom, the approach unifies many other model reduction techniques, such as singular limits, equilibrium approximations, and the renormalization group, while expanding the domain of tractable models. The method produces a series of approximations that decrease the complexity of the model and reveal how microscopic parameters are systematically "compressed" into a few macroscopic degrees of freedom, effectively building a bridge between the microscopic and the macroscopic descriptions.

  18. The Airframe Noise Reduction Challenge

    NASA Technical Reports Server (NTRS)

    Lockard, David P.; Lilley, Geoffrey M.

    2004-01-01

    The NASA goal of reducing external aircraft noise by 10 dB in the near-term presents the acoustics community with an enormous challenge. This report identifies technologies with the greatest potential to reduce airframe noise. Acoustic and aerodynamic effects will be discussed, along with the likelihood of industry accepting and implementing the different technologies. We investigate the lower bound, defined as noise generated by an aircraft modified with a virtual retrofit capable of eliminating all noise associated with the high lift system and landing gear. However, the airframe noise of an aircraft in this 'clean' configuration would only be about 8 dB quieter on approach than current civil transports. To achieve the NASA goal of 10 dB noise reduction will require that additional noise sources be addressed. Research shows that energy in the turbulent boundary layer of a wing is scattered as it crosses trailing edge. Noise generated by scattering is the dominant noise mechanism on an aircraft flying in the clean configuration. Eliminating scattering would require changes to much of the aircraft, and practical reduction devices have yet to receive serious attention. Evidence suggests that to meet NASA goals in civil aviation noise reduction, we need to employ emerging technologies and improve landing procedures; modified landing patterns and zoning restrictions could help alleviate aircraft noise in communities close to airports.

  19. The reduction of packaging waste

    SciTech Connect

    Raney, E.A.; Hogan, J.J.; McCollom, M.L.; Meyer, R.J.

    1994-04-01

    Nationwide, packaging waste comprises approximately one-third of the waste disposed in sanitary landfills. the US Department of Energy (DOE) generated close to 90,000 metric tons of sanitary waste. With roughly one-third of that being packaging waste, approximately 30,000 metric tons are generated per year. The purpose of the Reduction of Packaging Waste project was to investigate opportunities to reduce this packaging waste through source reduction and recycling. The project was divided into three areas: procurement, onsite packaging and distribution, and recycling. Waste minimization opportunities were identified and investigated within each area, several of which were chosen for further study and small-scale testing at the Hanford Site. Test results, were compiled into five ``how-to`` recipes for implementation at other sites. The subject of the recipes are as follows: (1) Vendor Participation Program; (2) Reusable Containers System; (3) Shrink-wrap System -- Plastic and Corrugated Cardboard Waste Reduction; (4) Cardboard Recycling ; and (5) Wood Recycling.

  20. Measuring Substantial Reductions in Activity

    PubMed Central

    Schafer, Charles; Evans, Meredyth; Jason, Leonard A.; So, Suzanna; Brown, Abigail

    2015-01-01

    The case definitions for Myalgic Encephalomyelitis/chronic fatigue syndrome (ME/CFS), Myalgic Encephalomyelitis (ME), and chronic fatigue syndrome (CFS) each include a disability criterion requiring substantial reductions in activity in order to meet diagnostic criteria. Difficulties have been encountered in defining and operationalizing the substantial reduction disability criterion within these various illness definitions. The present study sought to relate measures of past and current activities in several domains including the SF-36, an objective measure of activity (e.g. actigraphy), a self-reported quality of life scale, and measures of symptom severity. Results of the study revealed that current work activities had the highest number of significant associations with domains such as the SF-36 subscales, actigraphy, and symptom scores. As an example, higher self-reported levels of current work activity were associated with better health. This suggests that current work related activities may provide a useful domain for helping operationalize the construct of substantial reductions in activity. PMID:25584524

  1. Model reduction of flexible manipulators

    NASA Astrophysics Data System (ADS)

    Yang, Jie; Xu, Yangsheng; Chen, C. S.

    1992-06-01

    Flexible manipulators can be characterized by a dynamic model with a large number of vibration modes, and the use of the model in the model-based control schemes requires reduction of model order. Balanced truncation is an effective method for model reduction of asymptotically stable systems by transforming the states to a coordinate system in which the controllability and observability Gramians are equal and diagonal, and eliminating the states which contribute weakly to the input-output map. An elastic flexible manipulator, however, is a marginally stable system and thus the balanced truncation method can not be directly applied. Herein, a method is presented of reducing the order of a marginally stable system based on the fact that translation transformations in the frequency domain preserve input-output properties of the system. The successful application is addressed of the method to model reduction of flexible manipulators with infinite-dimensional for finite-dimensional model. The method is also applicable for any other marginally stable model, such as elastic space trusswork and multi-dimensional space vehicle structure.

  2. Rotational Invariant Dimensionality Reduction Algorithms.

    PubMed

    Lai, Zhihui; Xu, Yong; Yang, Jian; Shen, Linlin; Zhang, David

    2016-06-30

    A common intrinsic limitation of the traditional subspace learning methods is the sensitivity to the outliers and the image variations of the object since they use the L₂ norm as the metric. In this paper, a series of methods based on the L₂,₁-norm are proposed for linear dimensionality reduction. Since the L₂,₁-norm based objective function is robust to the image variations, the proposed algorithms can perform robust image feature extraction for classification. We use different ideas to design different algorithms and obtain a unified rotational invariant (RI) dimensionality reduction framework, which extends the well-known graph embedding algorithm framework to a more generalized form. We provide the comprehensive analyses to show the essential properties of the proposed algorithm framework. This paper indicates that the optimization problems have global optimal solutions when all the orthogonal projections of the data space are computed and used. Experimental results on popular image datasets indicate that the proposed RI dimensionality reduction algorithms can obtain competitive performance compared with the previous L₂ norm based subspace learning algorithms.

  3. Portland outlines waste reduction program

    SciTech Connect

    Rifer, W.

    1986-09-01

    Plans for solid waste systems must reflect the preferences and values of the community they serve. The Metropolitan Service District (Metro), an elected regional government for the Portland (Oregon) area, has launched an ambitious effort to reduce the amount of solid waste being landfilled. The current solid waste system in the Portland area consists of: (1) a healthy private recycling program which is achieving a nearly 22% reduction of waste, (2) a private garbage collection system which is regulated by local governments (half franchised and half competitive), and (3) a regional disposal system operated by Metro which relies on landfilling. The system envisioned by the Metro Waste Reduction Program will add significant diversity to the disposal system, and will, by necessity, involve the collection, recycling and disposal systems in a coordinated partnership. The total volume of waste to be handled through the Waste Reduction Program is approximately 962,000 tons/year. Currently, 755,000 tons/year are disposed of in a landfill and the remainder is recycled through various methods.

  4. Dietary supplements in weight reduction.

    PubMed

    Dwyer, Johanna T; Allison, David B; Coates, Paul M

    2005-05-01

    We summarize evidence on the role of dietary supplements in weight reduction, with particular attention to their safety and benefits. Dietary supplements are used for two purposes in weight reduction: (a) providing nutrients that may be inadequate in calorie-restricted diets and (b) for their potential benefits in stimulating weight loss. The goal in planning weight-reduction diets is that total intake from food and supplements should meet recommended dietary allowance/adequate intake levels without greatly exceeding them for all nutrients, except energy. If nutrient amounts from food sources in the reducing diet fall short, dietary supplements containing a single nutrient/element or a multivitamin-mineral combination may be helpful. On hypocaloric diets, the addition of dietary supplements providing nutrients at a level equal to or below recommended dietary allowance/adequate intake levels or 100% daily value, as stated in a supplement's facts box on the label, may help dieters to achieve nutrient adequacy and maintain electrolyte balance while avoiding the risk of excessive nutrient intakes. Many botanical and other types of dietary supplements are purported to be useful for stimulating or enhancing weight loss. Evidence of their efficacy in stimulating weight loss is inconclusive at present. Although there are few examples of safety concerns related to products that are legal and on the market for this purpose, there is also a paucity of evidence on safety for this intended use. Ephedra and ephedrine-containing supplements, with or without caffeine, have been singled out in recent alerts from the Food and Drug Administration because of safety concerns, and use of products containing these substances cannot be recommended. Dietitians should periodically check the Food and Drug Administration Web site ( www.cfsan.fda.gov ) for updates and warnings and alert patients/clients to safety concerns. Dietetics professionals should also consult authoritative sources for

  5. Tobacco, nicotine and harm reduction.

    PubMed

    Le Houezec, Jacques; McNeill, Ann; Britton, John

    2011-03-01

    Tobacco smoking, sustained by nicotine dependence, is a chronic relapsing disorder, which in many cases results in lifelong cigarette use and consequent death of one out of two lifelong smokers from a disease caused by their smoking. Most toxicity due to cigarette smoking is related to the burning process. Models of harm reduction applied to tobacco suggest that use of non-combustible, less toxic, nicotine-containing products as a substitute for cigarette smoking would reduce the death toll arising from tobacco use. Available options include medicinal nicotine and smokeless tobacco products. The potential role of nicotine replacement therapy (NRT) products in a harm reduction strategy is currently severely restricted by strict regulations on dose, safety and potential addictiveness. As a result, NRT products are designed to provide much less nicotine, and deliver it to the brain more slowly, than cigarettes, which are widely accessible and poorly regulated. Smokeless tobacco (snus) has proved to be an acceptable reduced hazard alternative to smoking in Sweden, but supply of snus is illegal elsewhere in the European Union. To increase accessibility and reach more smokers, barriers to the use of NRT use need to be removed and more effective NRTs need urgently to be developed. Smokeless tobacco could also play an important role in harm reduction, but current European Union regulations and concerns over exploitation by tobacco companies currently preclude wider use. To improve public health there is an urgent need for an appropriate regulatory framework and regulatory authority at the European level, controlling both tobacco and nicotine products to ensure that the least harmful products are the most accessible. © 2011 Australasian Professional Society on Alcohol and other Drugs.

  6. CCD data reductions at ESO

    NASA Astrophysics Data System (ADS)

    Grosbol, Preben

    The image-processing and data-reduction functions of the IHAP and MIDAS software packages developed at ESO for CCD astronomy are briefly reviewed. IHAP and MIDAS perform the same basic operations on HP 1000 and VAX computers, respectively, and MIDAS is currently being modified to run in the UNIX operating system as well as in VAX VMS. Consideration is given to the special properties of CCD data, the removal of gross errors (due to bad pixels and cosmic-ray events), photometric correction for dark current and sensitivity variations, digital filtering and Fourier transforms, detection and classification algorithms for direct imaging, surface photometry of extended objects, function fitting, and image deconvolution.

  7. XRT -- ROSAT XRT Data Reduction

    NASA Astrophysics Data System (ADS)

    Davenhall, A. C.; Platon, R. T.

    XRT is a package for reducing data acquired with the ROSAT XRT instruments. The XRT (X-Ray Telescope) was the principal scientific payload of the ROSAT X-ray astronomy satellite. The XRT had two instruments: the PSPC (Position Sensitive Proportional Counter) and the HRI (High Resolution Imager). The XRT package operates on data produced by these instruments and can be used to transform them into calibrated images, spectra, time-series etc. XRT was created by taking the ROSAT XRT-specific functions in the ASTERIX general X-ray astronomy data reduction system and re-packaging them as stand-alone applications.

  8. Selective reduction of heavy metals

    SciTech Connect

    Bjorling, G.

    1984-12-11

    The present invention relates to selective reduction of heavy metals out of finey grained, substantially oxidic material by blowing the oxidic material into a furnace together with an amount of reducing agent required for obtaining desired selectivity while simultaneously heat energy is supplied by a gas heated in a plasma generator, the temperature being adjusted to such a level as to correspond to the oxygen potential at which the desired metals are transformed into a particular, isolatable phase as metal melt, metal vapor, speiss or matte and at which the remaining metals enter into a slag phase and can be isolated as slag melt.

  9. Structural dynamics system model reduction

    NASA Technical Reports Server (NTRS)

    Chen, J. C.; Rose, T. L.; Wada, B. K.

    1987-01-01

    Loads analysis for structural dynamic systems is usually performed by finite element models. Because of the complexity of the structural system, the model contains large number of degree-of-freedom. The large model is necessary since details of the stress, loads and responses due to mission environments are computed. However, a simplified model is needed for other tasks such as pre-test analysis for modal testing, and control-structural interaction studies. A systematic method of model reduction for modal test analysis is presented. Perhaps it will be of some help in developing a simplified model for the control studies.

  10. Digital Noise Reduction: An Overview

    PubMed Central

    Bentler, Ruth; Chiou, Li-Kuei

    2006-01-01

    Digital noise reduction schemes are being used in most hearing aids currently marketed. Unlike the earlier analog schemes, these manufacturer-specific algorithms are developed to acoustically analyze the incoming signal and alter the gain/output characteristics according to their predetermined rules. Although most are modulation-based schemes (ie, differentiating speech from noise based on temporal characteristics), spectral subtraction techniques are being applied as well. The purpose of this article is to overview these schemes in terms of their differences and similarities. PMID:16959731

  11. Reductive Elimination from Arylpalladium Cyanide Complexes

    PubMed Central

    Klinkenberg, Jessica L.; Hartwig, John F.

    2012-01-01

    We report the isolation and characterization of arylpalladium cyanide complexes that undergo reductive elimination to form arylnitriles. The rates of reductive elimination from a series of arylpalladium cyanide complexes reveal that the electronic effects on the reductive elimination from arylpalladium cyanide complexes are distinct from those on reductive reductive eliminations from arylpalladium alkoxo, amido, thiolate, and enolate complexes. Arylpalladium cyanide complexes containing aryl ligands with electron-donating substituents undergo faster reductive elimination of aromatic nitriles than complexes containing aryl ligands with electron-withdrawing substituents. In addition, the transition state for the reductive elimination of the aromatic nitrile is much different from that for reductive eliminations that occur from most other arylpalladium complexes. Computational studies indicate that the reductive elimination of an arylnitrile from Pd(II) occurs through a transition state more closely related in structure and electronic distribution to that for the insertion of CO into a palladiumaryl bond. PMID:22352451

  12. Equivalent beam modeling using numerical reduction techniques

    NASA Technical Reports Server (NTRS)

    Chapman, J. M.; Shaw, F. H.

    1987-01-01

    Numerical procedures that can accomplish model reductions for space trusses were developed. Three techniques are presented that can be implemented using current capabilities within NASTRAN. The proposed techniques accomplish their model reductions numerically through use of NASTRAN structural analyses and as such are termed numerical in contrast to the previously developed analytical techniques. Numerical procedures are developed that permit reductions of large truss models containing full modeling detail of the truss and its joints. Three techniques are presented that accomplish these model reductions with various levels of structural accuracy. These numerical techniques are designated as equivalent beam, truss element reduction, and post-assembly reduction methods. These techniques are discussed in detail.

  13. Reduction of nitrate in Shewanella

    SciTech Connect

    Gao, Haichun; Yang, Zamin Koo; Barua, Sumitra; Reed, SB; Nealson, Kenneth H.; Fredrikson, JK; Tiedje, James; Zhou, Jizhong

    2009-01-01

    In the genome of Shewanella oneidensis, a napDAGHB gene cluster encoding periplasmic nitrate reductase (NapA) and accessory proteins and an nrfA gene encoding periplasmic nitrite reductase (NrfA) have been identified. These two systems seem to be atypical because the genome lacks genes encoding cytoplasmic membrane electron transport proteins, NapC for NAP and NrfBCD/NrfH for NRF, respectively. Here, we present evidence that reduction of nitrate to ammonium in S. oneidensis is carried out by these atypical systems in a two-step manner. Transcriptional and mutational analyses suggest that CymA, a cytoplasmic membrane electron transport protein, is likely to be the functional replacement of both NapC and NrfH in S. oneidensis. Surprisingly, a strain devoid of napB encoding the small subunit of nitrate reductase exhibited the maximum cell density sooner than the wild type. Further characterization of this strain showed that nitrite was not detected as a free intermediate in its culture and NapB provides a fitness gain for S. oneidensis to compete for nitrate in the environments. On the basis results from mutational analyses of napA, napB, nrfA and napBnrfA in-frame deletion mutants, we propose that NapB is able to favor nitrate reduction by routing electrons to NapA exclusively.

  14. Transport suppression by shear reduction

    NASA Astrophysics Data System (ADS)

    Martinell, Julio; Del-Castillo-Negrete, Diego

    2009-11-01

    The relationship between transport and shear is a problem of considerable interest to magnetically confined plasmas. It is well known that there are cases in which an increase of flow shear can lead to a reduction of turbulent transport. However, this is not a generic result, and there are transport problems in which the opposite is the case. In particular, as originally discussed in Ref. footnotetextdel-Castillo-Negrete and Morrison, Phys. Fluids A 5, 948 (1993), barriers to chaotic transport typically form in regions of vanishing shear. This property, which is generic to the so-called non-twist Hamiltonian systems footnotetextdel-Castillo-Negrete, Greene, and Morrison, Physica D 91, 1 (1996), explains the observed resilience of transport barriers in non-monotonic zonal flows in plasmas and fluids and the robustness of shearless magnetic surfaces in reverse shear configurations. Here we study the role of finite Larmor radius (FLR) effects on the suppression of chaotic transport by shear reduction in a simplified model. Following Ref. footnotetextdel-Castillo-Negrete, Phys. Plasmas, 7, 1702 (2000) we consider a model consisting of a superposition of drift waves and a non-monotonic zonal flow. The FLR effects are incorporated by gyroaveraging the E xB velocity, and transport is studied by following the evolution of ensembles of test particles.

  15. Biocatalytic reduction of carboxylic acids.

    PubMed

    Napora-Wijata, Kamila; Strohmeier, Gernot A; Winkler, Margit

    2014-06-01

    An increasing demand for non-petroleum-based products is envisaged in the near future. Carboxylic acids such as citric acid, succinic acid, fatty acids, and many others are available in abundance from renewable resources and they could serve as economic precursors for bio-based products such as polymers, aldehyde building blocks, and alcohols. However, we are confronted with the problem that carboxylic acid reduction requires a high level of energy for activation due to the carboxylate's thermodynamic stability. Catalytic processes are scarce and often their chemoselectivity is insufficient. This review points at bio-alternatives: currently known enzyme classes and organisms that catalyze the reduction of carboxylic acids are summarized. Two totally distinct biocatalyst lines have evolved to catalyze the same reaction: aldehyde oxidoreductases from anaerobic bacteria and archea, and carboxylate reductases from aerobic sources such as bacteria, fungi, and plants. The majority of these enzymes remain to be identified and isolated from their natural background in order to evaluate their potential as industrial biocatalysts.

  16. Supersonic jet shock noise reduction

    NASA Technical Reports Server (NTRS)

    Stone, J. R.

    1984-01-01

    Shock-cell noise is identified to be a potentially significant problem for advanced supersonic aircraft at takeoff. Therefore NASA conducted fundamental studies of the phenomena involved and model-scale experiments aimed at developing means of noise reduction. The results of a series of studies conducted to determine means by which supersonic jet shock noise can be reduced to acceptable levels for advanced supersonic cruise aircraft are reviewed. Theoretical studies were conducted on the shock associated noise of supersonic jets from convergent-divergent (C-D) nozzles. Laboratory studies were conducted on the influence of narrowband shock screech on broadband noise and on means of screech reduction. The usefulness of C-D nozzle passages was investigated at model scale for single-stream and dual-stream nozzles. The effect of off-design pressure ratio was determined under static and simulated flight conditions for jet temperatures up to 960 K. Annular and coannular flow passages with center plugs and multi-element suppressor nozzles were evaluated, and the effect of plug tip geometry was established. In addition to the far-field acoustic data, mean and turbulent velocity distributions were measured with a laser velocimeter, and shadowgraph images of the flow field were obtained.

  17. Double shrinking sparse dimension reduction.

    PubMed

    Zhou, Tianyi; Tao, Dacheng

    2013-01-01

    Learning tasks such as classification and clustering usually perform better and cost less (time and space) on compressed representations than on the original data. Previous works mainly compress data via dimension reduction. In this paper, we propose "double shrinking" to compress image data on both dimensionality and cardinality via building either sparse low-dimensional representations or a sparse projection matrix for dimension reduction. We formulate a double shrinking model (DSM) as an l(1) regularized variance maximization with constraint ||x||(2)=1, and develop a double shrinking algorithm (DSA) to optimize DSM. DSA is a path-following algorithm that can build the whole solution path of locally optimal solutions of different sparse levels. Each solution on the path is a "warm start" for searching the next sparser one. In each iteration of DSA, the direction, the step size, and the Lagrangian multiplier are deduced from the Karush-Kuhn-Tucker conditions. The magnitudes of trivial variables are shrunk and the importances of critical variables are simultaneously augmented along the selected direction with the determined step length. Double shrinking can be applied to manifold learning and feature selections for better interpretation of features, and can be combined with classification and clustering to boost their performance. The experimental results suggest that double shrinking produces efficient and effective data compression.

  18. Conformal optics risk reduction demonstration

    NASA Astrophysics Data System (ADS)

    Knapp, David J.; Mills, James P.; Hegg, Ronald G.; Trotta, Patrick A.; Smith, Christopher B.

    2001-09-01

    For the past three years, the Precision Conformal Optics Consortium has been developing a revolutionary new class of optics. These optics are characterized by outer window elements that conform to aerodynamic rather than optical requirements. Conformal optical elements can greatly improve the aerodynamic performance of the host platform. To make conformal optics a reality, challenges had to be overcome in design, fabrication, and testing. This was accomplished in October 1999 when Raytheon demonstrated the world's first conformal optical system. This fineness ratio one system was a risk reduction effort to develop technology for later systems. It is comprised of a calcium fluoride conformal optical dome, a TI-1173 aspheric corrector, and a calcium fluoride solid catadioptric telescope. The design involved overcoming large amounts of aberration that varied with gimbal look angle. Efforts also included aligning the system to tight tolerances and testing highly aspheric optical elements. Overall, the actual system performance compared very favorably with the design model. With the proven success of this risk reduction demonstration, the path was cleared for new higher performance conformal optical systems.

  19. NOX REMOVAL WITH COMBINED SELECTIVE CATALYTIC REDUCTION AND SELECTIVE NONCATALYTIC REDUCTION: PILOT- SCALE TEST RESULTS

    EPA Science Inventory

    Pilot-scale tests were conducted to develop a combined nitrogen oxide (NOx) reduction technology using both selective catalytic reduction (SCR) and selective noncatalytic reduction (SNCR). A commercially available vanadium-and titatnium-based composite honeycomb catalyst and enh...

  20. NOX REMOVAL WITH COMBINED SELECTIVE CATALYTIC REDUCTION AND SELECTIVE NONCATALYTIC REDUCTION: PILOT- SCALE TEST RESULTS

    EPA Science Inventory

    Pilot-scale tests were conducted to develop a combined nitrogen oxide (NOx) reduction technology using both selective catalytic reduction (SCR) and selective noncatalytic reduction (SNCR). A commercially available vanadium-and titatnium-based composite honeycomb catalyst and enh...