Microtubules do not promote mitotic slippage when the spindle assembly checkpoint cannot be satisfied

© 2008 Brito et al. This article is distributed under the terms of the Creative Commons Attribution 3.0 License. The definitive version was published in Journal of Cell Biology 182 (2009): 623-629, doi:10.1083/jcb.200805072.When the spindle assembly checkpoint (SAC) cannot be satisfied, cells exit mitosis via mitotic slippage. In microtubule (MT) poisons, slippage requires cyclin B proteolysis, and it appears to be accelerated in drug concentrations that allow some MT assembly. To determine if MTs accelerate slippage, we followed mitosis in human RPE-1 cells exposed to various spindle poisons. At 37°C, the duration of mitosis in nocodazole, colcemid, or vinblastine concentrations that inhibit MT assembly varied from 20 to 30 h, revealing that different MT poisons differentially depress the cyclin B destruction rate during slippage. The duration of mitosis in Eg5 inhibitors, which induce monopolar spindles without disrupting MT dynamics, was the same as in cells lacking MTs. Thus, in the presence of numerous unattached kinetochores, MTs do not accelerate slippage. Finally, compared with cells lacking MTs, exit from mitosis is accelerated over a range of spindle poison concentrations that allow MT assembly because the SAC becomes satisfied on abnormal spindles and not because slippage is accelerated.This research was supported by The National Institutes of Health (GMS 40198 to C.L. Rieder) and a doctoral research fellowship (SFRH/ BD/13663/2003) from Fundacao para a Ciencia e a Tecnologia (to D.A. Brito)

Cells satisfy the mitotic checkpoint in Taxol, and do so faster in concentrations that stabilize syntelic attachments

© The Authors, 2009 . This article is distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License. The definitive version was published in Journal of Cell Biology 186 (2009): 675-684, doi:10.1083/jcb.200906150.To determine why the duration of mitosis (DM) is less in Taxol than in nocodazole or Eg5 inhibitors we studied the relationship between Taxol concentration, the DM, and the mitotic checkpoint. We found that unlike for other spindle poisons, in Taxol the DM becomes progressively shorter as the concentration surpasses ~0.5 µM. Studies on RPE1 and PtK2 expressing GFP/cyclin B or YFP/Mad2 revealed that cells ultimately satisfy the checkpoint in Taxol and do so faster at concentrations >0.5 µM. Inhibiting the aurora-B kinase in Taxol-treated RPE1 cells accelerates checkpoint satisfaction by stabilizing syntelic kinetochore attachments and reduces the DM to ~1.5 h regardless of drug concentration. A similar stabilization of syntelic attachments by Taxol itself appears responsible for accelerated checkpoint satisfaction at concentrations >0.5 µM. Our results provide a novel conceptual framework for how Taxol prolongs mitosis and caution against using it in checkpoint studies. They also offer an explanation for why some cells are more sensitive to lower versus higher Taxol concentrations.This work was supported by National Institutes of Health GMS grant 40198 to C.L. Rieder

Nudel functions in membrane traffic mainly through association with Lis1 and cytoplasmic dynein

Nudel and Lis1 appear to regulate cytoplasmic dynein in neuronal migration and mitosis through direct interactions. However, whether or not they regulate other functions of dynein remains elusive. Herein, overexpression of a Nudel mutant defective in association with either Lis1 or dynein heavy chain is shown to cause dispersions of membranous organelles whose trafficking depends on dynein. In contrast, the wild-type Nudel and the double mutant that binds to neither protein are much less effective. Time-lapse microscopy for lysosomes reveals significant reduction in both frequencies and velocities of their minus end–directed motions in cells expressing the dynein-binding defective mutant, whereas neither the durations of movement nor the plus end–directed motility is considerably altered. Moreover, silencing Nudel expression by RNA interference results in Golgi apparatus fragmentation and cell death. Together, it is concluded that Nudel is critical for dynein motor activity in membrane transport and possibly other cellular activities through interactions with both Lis1 and dynein heavy chain

Influence of a groove-structured vortex generator on the drag reduction characteristics of a multiphase pump

The oil–gas mixture pump significantly contributes to marginal oil field extraction and remote transportation of deep-sea oil. Nevertheless, during the operation of the mixture pump, it is inevitable to encounter problems like the separation of the mixed media from the hydraulic components as well as the gas phase from the liquid phase, which leads to enhancing the flow resistance of the mixed media. Therefore, this study investigates the influence of a groove-structure vortex generator on the drag reduction characteristics of a helical axial-flow gas–liquid multiphase pump under the design flow rate condition and various inlet gas content rates. The findings show that the vortex generator with diverse groove depths can prevent the separation of the mixed media from the blade suction surface effectively and minimize the flow resistance of the media in the 1/10 of the blade inlet. In particular, excellent drag reduction results were gained with a maximum drag reduction rate of 36.7% when the relative depth was 3/40. In addition, the efficiency of the mixture pump increased by a maximum of 2.1%, and the head increased by a maximum of 4.3%. The significance of this study lies in its potential to further optimize the design and performance of gas–liquid multiphase pumps. It provides new insights into the design and application of vortex generators. It offers robust support for the optimization and enhancement of gas–liquid multiphase pumps

Development of an efficient numerical method for wind turbine flow, sound generation, and propagation under multi-wake conditions

The propagation of aerodynamic noise from multi-wind turbines is studied. An efficient hybrid method is developed to jointly predict the aerodynamic and aeroacoustics performances of wind turbines, such as blade loading, rotor power, rotor aerodynamic noise sources, and propagation of noise. This numerical method combined the simulations of wind turbine flow, noise source and its propagation which is solved for long propagation path and under complex flow environment. The results from computational fluid dynamics (CFD) calculations not only provide wind turbine power and thrust information, but also provide detailed wake flow. The wake flow is computed with a 2D actuator disc (AD) method that is based on the axisymmetric flow assumption. The relative inflow velocity and angle of attack (AOA) of each blade element form input data to the noise source model. The noise source is also the initial condition for the wave equation that solves long distance noise propagation in frequency domain. Simulations were conducted under different atmospheric conditions which showed that wake flow is an important part that has to be included in wind turbine noise propagation

Extra centrosomes and/or chromosomes prolong mitosis in human cells

Author Posting. © The Author(s), 2008. This is the author's version of the work. It is posted here by permission of Nature Publishing Group for personal use, not for redistribution. The definitive version was published in Nature Cell Biology 10 (2008): 748-751, doi:10.1038/ncb1738.Using laser microsurgery and cell fusion we have explored how additional centrosomes and/or chromosomes influence the duration of mitosis in human cells. We find that doubling the chromosome number adds ~10 minutes to a 20 minute division while doubling the number of centrosomes adds ~30 minutes more, and extra centrosomes and/or chromosomes prolong mitosis by delaying satisfaction of the spindle assembly checkpoint. Thus mitosis can be prolonged by non genetic means and extra chromosomes and centrosomes likely contribute to the elevated mitotic index seen in many tumors.This work was supported by National Institutes of General Medical Sciences grants 40198 (to C.L.R.) and 59363 (to A.K.)

A Study on the Oil-Bearing Stability of Salt-Resistant Foam and an Explanation of the Viscoelastic Phenomenon

Foam is a medium-stable system composed of gas and liquid phases, which has the advantages of low density at the gas phase and high viscosity at the liquid phase, and has a wide application in oil and gas field development and mineral flotation, but its special medium-stable system also brings many problems in industry applications. Scientists have carried out extensive analyses and research on the foam stability and bubble-bursting mechanism, which initially clarified the rules of bubble breakage caused by environmental factors such as temperature and pressure, but the mechanism of bubble bursting under the action of internal factors such as liquid mineralization and oil concentration of the films is still not clearly defined. In this paper, we propose a compound salt-resistant foaming agent, investigated the influence of the aggregation and adsorption behavior of oil droplets on the liquid films and boundaries, and established a relevant aggregation and adsorption model with the population balance equation. We put forward a liquid film drainage mechanism based on the distribution, aggregation, and transport of oil droplets in the liquid films, so as to explain the changes in foam stability under the action of oil droplets. On the other hand, the viscoelastic analysis of foam fluid is performed with a rheometer, and the results show that in comparison with conventional power-law fluid, foam fluid has a complex rheological behavior for low shear thickening, but high shear thinning