scholarly journals Kinetochore motility after severing between sister centromeres using laser microsurgery: evidence that kinetochore directional instability and position is regulated by tension

1995 ◽  
Vol 108 (7) ◽  
pp. 2537-2548 ◽  
Author(s):  
R.V. Skibbens ◽  
C.L. Rieder ◽  
E.D. Salmon
Keyword(s):  
Laser Ablation ◽  
Constant Velocity ◽  
Somatic Cells ◽  
Metaphase Plate ◽  
Fundamental Property ◽  
The Other ◽  
Centromeric Chromatin ◽  
Laser Microbeam ◽  
Differential Movement ◽  
Sister Kinetochore

During mitosis in vertebrate somatic cells, the single attached kinetochore on a mono-oriented chromosome exhibits directional instability: abruptly and independently switching between constant velocity poleward and away from the pole motility states. When the non-attached sister becomes attached to the spindle (chromosome bi-orientation), the motility of the sister kinetochores becomes highly coordinated, one moving poleward while the other moves away from the pole, allowing chromosomes to congress to the spindle equator. In our kinetochore-tensiometer model, we hypothesized that this coordinated behavior is regulated by tension across the centromere produced by kinetochore movement relative to the sister kinetochore and bulk of the chromosome arms. To test this model, we severed or severely weakened the centromeric chromatin between sister kinetochores on bi-oriented newt lung cell chromosomes with a laser microbeam. This procedure converted a pair of tightly linked sister kinetochores into two mono-oriented single kinetochore-chromatin fragments that were tethered to their chromosome arms by thin compliant chromatin strands. These single kinetochore-chromatin fragments moved substantial distances off the metaphase plate, stretching their chromatin strands, before the durations of poleward and away from the pole movement again became similar. In contrast, the severed arms remained at or moved closer to the spindle equator. The poleward and away from the pole velocities of single kinetochore-chromatin fragments in prometaphase were typical of velocities exhibited by sister kinetochores on intact chromosomes from prometaphase through midanaphase A. However, severing the chromatin between sister kinetochores uncoupled the normally coordinated motility of sister kinetochores. Laser ablation also uncoupled the motilities of the single kinetochore fragments from the bulk of the arms. These results reveal that kinetochore directional instability is a fundamental property of the kinetochore and that the motilities of sister kinetochores are coordinated during congression by a stiff centromere linkage. We conclude that kinetochores act as tensiometers that sense centromere tension generated by differential movement of sister kinetochores and their chromosome arms to control switching between constant velocity P and AP motility states.

scholarly journals Condensin Regulates the Stiffness of Vertebrate Centromeres

2009 ◽  
Vol 20 (9) ◽  
pp. 2371-2380 ◽  
Author(s):  
Susana A. Ribeiro ◽  
Jesse C. Gatlin ◽  
Yimin Dong ◽  
Ajit Joglekar ◽  
Lisa Cameron ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Metaphase Plate ◽  
Centromeric Chromatin ◽  
Anaphase Onset ◽  
Sister Kinetochore ◽  
Pulling Forces ◽  
Important Regulator ◽  
Chromatin Folding ◽  
And Function ◽  
Rest Length

When chromosomes are aligned and bioriented at metaphase, the elastic stretch of centromeric chromatin opposes pulling forces exerted on sister kinetochores by the mitotic spindle. Here we show that condensin ATPase activity is an important regulator of centromere stiffness and function. Condensin depletion decreases the stiffness of centromeric chromatin by 50% when pulling forces are applied to kinetochores. However, condensin is dispensable for the normal level of compaction (rest length) of centromeres, which probably depends on other factors that control higher-order chromatin folding. Kinetochores also do not require condensin for their structure or motility. Loss of stiffness caused by condensin-depletion produces abnormal uncoordinated sister kinetochore movements, leads to an increase in Mad2(+) kinetochores near the metaphase plate and delays anaphase onset.

scholarly journals Microtubule assembly and kinetochore directional instability in vertebrate monopolar spindles: implications for the mechanism of chromosome congression

1994 ◽  
Vol 107 (1) ◽  
pp. 285-297 ◽  
Author(s):  
L. Cassimeris ◽  
C.L. Rieder ◽  
E.D. Salmon
Keyword(s):  
Constant Velocity ◽  
Metaphase Plate ◽  
Lung Epithelial Cells ◽  
Microtubule Assembly ◽  
Pole Motion ◽  
Chromosome Congression ◽  
Chromosome Position ◽  
Lung Epithelial ◽  
Sister Kinetochore ◽  
Bipolar Spindles

We have proposed previously a kinetochore motor-polar ejection model for chromosome congression to the metaphase plate where forces generated at the kinetochore are antagonized by away-from-the pole forces generated within each half-spindle on the chromosome arms. This model was based in large part on observations of the behavior of chromosomes on monopolar spindles. In these cells chromosomes typically become attached to the pole by only one kinetochore fiber. These mono-oriented chromosomes move to positions away from the pole even though they are pulled poleward at their kinetochores. Their arms are also ejected away from the pole when severed from the centromere. Here we have characterized further the properties of monopolar spindles in newt lung epithelial cells to determine the similarities between monopolar and bipolar spindles. We found no significant differences between monopolar and bipolar spindles over the parameters examined, which included: microtubule dynamics as measured by fluorescence redistribution after photobleaching; the ability of polar microtubule arrays to push chromosome arms away from the pole; the dependence of chromosome position relative to the pole on microtubule assembly; the number of kinetochore microtubules per kinetochore; and the directional instability of kinetochore motion during chromosome oscillations poleward and away-from-the-pole. As in bipolar spindles, kinetochore directional instability is characterized by abrupt switching between constant velocity phases of poleward and away-from-the-pole motion. From these data we conclude that the mechanism(s) responsible for chromosome positioning in monopolar spindles are fundamentally the same as those in bipolar spindles; only the geometry of the two spindle forms and the interplay between sister kinetochore directional instabilities are different. We also found no correlation in the kinetochore-to-pole distance with kinetochore microtubule number in monopolar spindles, but a strong qualitative correlation with microtubule density. This finding indicates that oscillations of mono-oriented chromosomes in both monopolar and bipolar spindles occur because chromosomes persist in poleward motion until they reach a density of polar microtubules sufficiently high to promote switching to away-from-the-pole motion. As the kinetochore and chromosome arms move away-from-the-pole, microtubule density decreases and the kinetochore switches to poleward motion, pulling the chromosome arms back into regions of higher microtubule density. The mechanism regulating kinetochore switching between poleward and away-from-the-pole motion is poorly understood, but may depend on tension at the kinetochore generated by pushing forces on the chromosome arms produced by the polar microtubule arrays.

Regulation And Effect Of Telomerase And Telomeric Length In Stem Cells

2020 ◽  
Vol 15 ◽  
Author(s):  
Basak Celtikci ◽  
Gulnihal Kulaksiz Erkmen ◽  
Zeliha Gunnur Dikmen
Keyword(s):  
Telomere Length ◽  
Somatic Cells ◽  
Telomere Maintenance ◽  
The Other ◽  
Telomerase Reverse Transcriptase ◽  
Human Telomerase Reverse Transcriptase ◽  
Telomere Shortening ◽  
Maintenance Mechanism ◽  
Associated Diseases ◽  
Human Telomerase

: Telomeres are the protective end caps of eukaryotic chromosomes and they decide the proliferative lifespan of somatic cells, as the guardians of the cell replication. Telomere length in leucocytes reflects telomere length in other somatic cells. Leucocyte telomere length can be a biomarker of human ageing. The risk of diseases, which are associated with reduced cell proliferation and tissue degeneration, including aging or aging-associated diseases, such as dyskeratosis congenita, cardiovascular diseases, pulmonary fibrosis and aplastic anemia, are correlated with an increase in short telomeres. On the other hand, the risk of diseases, which are associated with increased proliferative growth, including major cancers, is correlated with long telomeres. In most of the cancers, a telomere maintenance mechanism during DNA replication is essential. The reactivation of the functional ribonucleoprotein holoenzyme complex [telomerase] starts the cascade from normal and premalignant somatic cells to advanced malignant cells. Telomerase is overexpressed during the development of cancer and embryonic stem cells, through controlling genome integrity, cancer formation and stemness. Cancer cells have mechanisms to maintain telomeres to avoid initiation of cellular senescence or apoptosis, and halting cell division by critically short telomeres. Modulation of the human telomerase reverse transcriptase is the ratelimiting step for the production of functional telomerase and the telomere maintenance. Human telomerase reverse transcriptase promoter promotes its gene expression only in tumor cells, but not in normal cells. Some cancers activate an alternative lengthening of telomeres maintenance mechanism via DNA recombination to unshorten their telomeres. Not only heritability but also oxidative stress, inflammation, environmental factors, and therapeutic interventions have an effect on telomere shortening, explaining the variability in telomere length across individuals. There have been a large number of publications, which correlate human diseases with progressive telomere shortening. Telomere length of an individual at birth is also important to follow up telomere shortening, and it can be used as biomarkers for healthy aging. On the other hand, understanding of cellular stress factors, which affect stem cell behavior, will be useful in regeneration or treatment in cancer and age-associated diseases. In this review, we will understand the connection between stem cell and telomere biology, cancer, and aging-associated diseases. This connection may be useful for discovering novel drug targets and improve outcomes for patients having cancer and aging-associated diseases.

Using Low Temperature, Cost-effective and Green Methods to Carbon Nanohorn Synthesis

2019 ◽  
Vol 9 (2) ◽  
pp. 157-160
Author(s):  
Ali Hasani
Keyword(s):  
Laser Ablation ◽  
Melting Point ◽  
Palm Olein ◽  
Arc Discharge ◽  
Low Cost ◽  
The Other ◽  
High Yield ◽  
Reaction Field ◽  
Carbon Nanohorns ◽  
Graphite Rod

Background: Laser ablation method has high-yield and pure SWCNHs. On the other hand, arc discharge methods have low-cost production of SWCNHs. However, these techniques have more desirable features, they need special expertness to use high power laser or high current discharge that either of them produces very high temperature. As for the researches, the temperatures of these techniques are higher than 4727°C to vaporize the graphite. So, to become aware of the advantages of SWCNHs, it is necessary to find a new way to synthesize SWCNHs at a lower temperature. In other words, reaction field can be expandable at a moderate temperature. This paper reports a new way to synthesize SWCNHs at an extremely reduced temperature. Methods: According to this study, the role of N2 is the protection of the copper holder supporting the graphite rod by increasing heat transfer from the holder. After the current of 70 A was supplied to the system, the temperature of graphite rod was raised to 1600°C. It is obvious that this temperature is somehow higher than the melting point of palladium, 1555°C, and much lower than graphite melting point, 3497°C. Results: Based on the results, there are transitional precursors simultaneous with the SWCNHs. This composition can be created by distortion of the primary SWCNTs at the higher temperature. Subsequently, each SWCNTs have a tendency to be broken into individual horns. With increasing the concentration of the free horns, bud-like SWCNHs can be produced. Moreover, there are individual horns almost separated from the mass of single wall carbon nanohorns. This structure is not common in SWCNHs synthesized by the usual method such as arc discharge or laser ablation. Through these regular techniques, SWCNHs are synthesized as cumulative particles with diameters about 30-150 nm. Conclusion: A simple heating is needed for SWCNTs transformation to SWCNHs with the presence of palladium as catalyst. The well-thought-out mechanism for this transformation is that SWCNTs were initially changed to highly curled shape, and after that were formed into small independent horns. The other rout to synthesize SWCNHs is the pyrolysis of palm olein at 950°C with the assistance of zinc nitrate and ferrocene. Palm olein was used as a promising, bio-renewable and inexpensive carbon source for the production of carbon nanohorns.

Development of the Quadripolar Meiotic Cytoskeleton in Spore Mother Cells of the Moss Funaria Hygrometrica

1988 ◽  
Vol 91 (1) ◽  
pp. 127-137
Author(s):  
C. H. BUSBY ◽  
B.E. S. GUNNING
Keyword(s):  
Meiotic Prophase ◽  
Metaphase Plate ◽  
Plant Cells ◽  
Preprophase Band ◽  
The Other ◽  
Higher Plant ◽  
Funaria Hygrometrica ◽  
Condensed Form ◽  
Prophase Nucleus ◽  
Mother Cells

Evidence presented in the accompanying paper that plastids function as microtubule (MT)-organizing centres for development of the quadripolar cytoskeleton of pre-meiotic spore mother cells (SMCs) in the moss Funaria hygrometrica is complemented here by observations on the MT system in these cells. Early in meiotic prophase numerous MTs align progressively along the two plastids as they elongate. Concomitant with (and perhaps causal for) plastid rotation, new MT arrays grow from each tip of each plastid to both tips of the other plastid. The ‘along-plastid’ and ‘between-plastid’ arrays ultimately form the edges of a tetrahedron, enclosing the prophase nucleus. MT breakdown at the centre of each edge leaves four cones of MTs, one emanating from each vertex, located at the plastid tips. These partially fuse in between-plastid pairs to give a twisted spindle with broad knife-edge poles oriented at right angles to one another, i.e. a condensed form of the quadripolar precursor. The twist causes the metaphase plate and the subsequent phragmoplast and organelle band to be saddle-shaped, and the daughter nuclei to be elongated perpendicular to one another along the two knife edges. The tetrahedral array returns during interkinesis and again breaks down into four cones of MTs centred on the plastid tips; these, however, now become individual half spindles for the two perpendicularly arranged second division spindles. When meiosis is completed the four haploid nuclei thus come to lie at the vertices of a tetrahedron that was established by MT-mediated plastid positioning during meiotic prophase. The tetrahedral cage of MTs precedes meiosis yet predicts the planes of division, and in these two respects it is the meiotic counterpart of the preprophase band of MTs, which develops before mitosis in most higher plant cells.

scholarly journals The influence of vortices upon the resitance experienced by solids moving through a liquid

1928 ◽  
Vol 119 (781) ◽  
pp. 146-156 ◽  
Keyword(s):  
Perfect Fluid ◽  
Relative Motion ◽  
Constant Velocity ◽  
Fluid Motion ◽  
Resultant Force ◽  
The Other ◽  
Two Dimensional ◽  
Perfect Fluids ◽  
Potential Problems ◽  
The Moment

(1) It is not so long ago that it was generally believed that the "classical" hydrodynamics, as dealing with perfect fluids, was, by reason of the very limitations implied in the term "perfect," incapable of explaining many of the observed facts of fluid motion. The paradox of d'Alembert, that a solid moving through a liquid with constant velocity experienced no resultant force, was in direct contradiction with the observed facts, and, among other things, made the lift on an aeroplane wing as difficult to explain as the drag. The work of Lanchester and Prandtl, however, showed that lift could be explained if there was "circulation" round the aerofoil. Of course, in a truly perfect fluid, this circulation could not be produced—it does need viscosity to originate it—but once produced, the lift follows from the theory appropriate to perfect fluids. It has thus been found possible to explain and calculate lift by means of the classical theory, viscosity only playing a significant part in the close neighbourhood ("grenzchicht") of the solid. It is proposed to show, in the present paper, how the presence of vortices in the fluid may cause a force to act on the solid, with a component in the line of motion, and so, at least partially, explain drag. It has long been realised that a body moving through a fluid sets up a train of eddies. The formation of these needs a supply of energy, ultimately dissipated by viscosity, which qualitatively explains the resistance experienced by the solid. It will be shown that the effect of these eddies is not confined to the moment of their birth, but that, so long as they exist, the resultant of the pressure on the solid does not vanish. This idea is not absolutely new; it appears in a recent paper by W. Müller. Müller uses some results due to M. Lagally, who calculates the resultant force on an immersed solid for a general fluid motion. The result, as far as it concerns vortices, contains their velocities relative to the solid. Despite this, the term — ½ ρq 2 only was used in the pressure equation, although the other term, ρ ∂Φ / ∂t , must exist on account of the motion. (There is, by Lagally's formulæ, no force without relative motion.) The analysis in the present paper was undertaken partly to supply this omission and partly to check the result of some work upon two-dimensional potential problems in general that it is hoped to publish shortly.

scholarly journals Using Kalman Filters to Reduce Noise from RFID Location System

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Pedro Henriques Abreu ◽  
José Xavier ◽  
Daniel Castro Silva ◽  
Luís Paulo Reis ◽  
Marcelo Petry
Keyword(s):  
Kalman Filter ◽  
Kalman Filters ◽  
Constant Velocity ◽  
Research Work ◽  
The Other ◽  
Full Potential ◽  
Comparison Study ◽  
Location System ◽  
Oval Shape ◽  
Location Systems

Nowadays, there are many technologies that support location systems involving intrusive and nonintrusive equipment and also varying in terms of precision, range, and cost. However, the developers some time neglect the noise introduced by these systems, which prevents these systems from reaching their full potential. Focused on this problem, in this research work a comparison study between three different filters was performed in order to reduce the noise introduced by a location system based on RFID UWB technology with an associated error of approximately 18 cm. To achieve this goal, a set of experiments was devised and executed using a miniature train moving at constant velocity in a scenario with two distinct shapes—linear and oval. Also, this train was equipped with a varying number of active tags. The obtained results proved that the Kalman Filter achieved better results when compared to the other two filters. Also, this filter increases the performance of the location system by 15% and 12% for the linear and oval paths respectively, when using one tag. For a multiple tags and oval shape similar results were obtained (11–13% of improvement).

DNA supercoiling and transcription: topological coupling of promoters

1996 ◽  
Vol 29 (3) ◽  
pp. 203-225 ◽  
Author(s):  
David M. J. Lilley ◽  
Dongrong Chen ◽  
Richard P. Bowater
Keyword(s):  
Fundamental Property ◽  
Dna Supercoiling ◽  
The Other ◽  
Linear Dna ◽  
Dna Molecule ◽  
Linking Number ◽  
Closed Circle ◽  
A Chain

DNA supercoiling is a consequence of the double-stranded nature of DNA. When a linear DNA molecule is ligated into a covalently closed circle, the two strands become intertwined like the links of a chain, and will remain so unless one of the strands is broken. The number of times one strand is linked with the other is described by a fundamental property of DNA supercoiling, the linking number (Lk).

scholarly journals The role of the centriolar region in animal cell mitosis. A laser microbeam study.

1977 ◽  
Vol 72 (2) ◽  
pp. 351-367 ◽  
Author(s):  
M W Berns ◽  
J B Rattner ◽  
S Brenner ◽  
S Meredith
Keyword(s):  
Animal Cell ◽  
Metaphase Plate ◽  
B Chromosome ◽  
Ultrastructural Analysis ◽  
Laser Microbeam ◽  
Membrane Breakdown ◽  
Pericentriolar Material ◽  
Ion Laser ◽  
Argon Ion

An argon ion laser microbeam (488 and 514 nm) was used to selectively irradiate one of the two centriolar regions of rat kangaroo Potorous tridactylis (PtK2) prophase cells in vitro. The cells were sensitized to the laser radiation by treatment with acridine orange (0.1-0.2 mug/ml). Ultrastructural examination of the irradiated centriolar regions demonstrated that the primary site of damage was the pericentriolar material. This result suggests that nucleic acid is present in the pericentriolar material. Behavioral and ultrastructural analysis demonstrated that cells with one damaged pericentriolar zone could undergo (a) nuclear membrane breakdown, (b) chromosome condensation, (c) metaphase plate formation, and (d) cytokinesis. However, the chromosomes neither separated nor exhibited any anaphase movements. Detailed ultrastructural analysis revealed the presence of kinetochore microtubules on both sides of chromosome mass and a lack of microtubules in the cytokinesis constriction. These results indicate that the pericentriolar material is important in spindle orgainization and essential for the formation of the interpolar microtubules.

Deburring Effect of Plasma Produced by Nanosecond Laser Ablation

2014 ◽  
Vol 136 (2) ◽  
Author(s):  
Yun Zhou ◽  
Yibo Gao ◽  
Benxin Wu ◽  
Sha Tao ◽  
Ze Liu
Keyword(s):  
Shock Wave ◽  
Laser Ablation ◽  
High Spatial Resolution ◽  
Shape Change ◽  
The Other ◽  
Nanosecond Laser ◽  
Laser Induced Plasma ◽  
Nanosecond Laser Ablation ◽  
Mechanical Breaking ◽  
The Sacrifice

This paper presents an interesting nanosecond (ns) laser-induced plasma deburring (LPD) effect (from microchannel sidewalls) discovered by the authors, which has been rarely reported before in the literature. Fast imagining study has been performed on plasma produced by ns laser ablation of the bottom of microchannels. It has been found that the plasma can effectively remove burrs from the sidewall of the channels, while on the other hand microscopic images taken in this study did not show any obvious size or shape change of the channel sidewall after LPD. LPD using a sacrifice plate has also been studied, where the plasma for deburring is generated by laser ablation of the sacrifice plate instead of the workpiece. The observed laser-induced plasma deburring effect has several potential advantages in practical micromanufacturing applications, such as high spatial resolution, noncontact and no tool wear, and less possibility of damaging or overmachining useful microfeatures when removing burrs from them. The fundamental mechanisms for the observed laser-induced plasma deburring effect still require lots of further work to completely understand, which may include mechanical breaking of burrs due to high kinetic energies carried by plasma and the associated shock wave, and/or thermal transport from plasma to burrs that may cause their heating and phase change, or other mechanisms.

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