Microtubule sliding activity of a kinesin-8 promotes spindle assembly and spindle length control

Molecular motors play critical roles in the formation of mitotic spindles, either through controlling the stability of individual microtubules, or by cross-linking and sliding microtubule arrays. Kinesin-8 motors are best known for their regulatory roles in controlling microtubule dynamics. They contain microtubule-destabilizing activities, and restrict spindle length in a wide variety of cell types and organisms. Here, we report for the first time on an anti-parallel microtubule-sliding activity of the budding yeast kinesin-8, Kip3. The in vivo importance of this sliding activity was established through the identification of complementary Kip3 mutants that separate the sliding activity and microtubule destabilizing activity. In conjunction with kinesin-5/Cin8, the sliding activity of Kip3 promotes bipolar spindle assembly and the maintenance of genome stability. We propose a “slide-disassemble” model where Kip3’s sliding and destabilizing activity balance during pre-anaphase. This facilitates normal spindle assembly. However, Kip3’s destabilizing activity dominates in late anaphase, inhibiting spindle elongation and ultimately promoting spindle disassembly

Nurses' perceptions of aids and obstacles to the provision of optimal end of life care in ICU

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MAP65/Ase1 promote microtubule flexibility.

International audienceMicrotubules (MTs) are dynamic cytoskeletal elements involved in numerous cellular processes. Although they are highly rigid polymers with a persistence length of 1-8 mm, they may exhibit a curved shape at a scale of few micrometers within cells, depending on their biological functions. However, how MT flexural rigidity in cells is regulated remains poorly understood. Here we ask whether MT-associated proteins (MAPs) could locally control the mechanical properties of MTs. We show that two major cross-linkers of the conserved MAP65/PRC1/Ase1 family drastically decrease MT rigidity. Their MT-binding domain mediates this effect. Remarkably, the softening effect of MAP65 observed on single MTs is maintained when MTs are cross-linked. By reconstituting physical collisions between growing MTs/MT bundles, we further show that the decrease in MT stiffness induced by MAP65 proteins is responsible for the sharp bending deformations observed in cells when they coalign at a steep angle to create bundles. Taken together, these data provide new insights into how MAP65, by modifying MT mechanical properties, may regulate the formation of complex MT arrays

Veno-arterial extracorporeal membrane oxygenation for drug intoxications: A single center, 14-year experience

International audienceBACKGROUND AND AIM OF THE STUDY: Acute cardiovascular failure remains a leading cause of death in severe poisonings. Veno-arterial extracorporeal membrane oxygenation (VA-ECMO) has been increasingly used as a rescue therapeutic option for those cases refractory to optimal conventional treatment. We sought to evaluate the outcomes after VA-ECMO used for drug intoxications in a single-center experience. METHODS: We performed an observational analysis of our prospective institutional database. The primary endpoint was survival to hospital discharge. RESULTS: Between January 2007 and December 2020, 32 patients (mean age: 45.4 ± 15.8 years; 62.5% female) received VA-ECMO for drug intoxication-induced refractory cardiogenic shock (n = 25) or cardiac arrest (n = 7). Seven (21.8%) patients developed lower limb ischemia during VA-ECMO support. Twenty-six (81.2%) patients were successfully weaned after a mean VA-ECMO support of 2.9 ± 1.3 days. One (3.1%) patient died after VA-ECMO weaning for multiorgan failure and survival to hospital discharge was 78.1% (n = 25). In-hospital survivors were discharged from hospital with a good neurological status. Survival to hospital discharge was not statistically different according to sex (male = 75.0% vs. female = 80.0%; p = .535), type of intoxication (single drug = 81.8% vs. multiple drugs = 76.1%; p = .544) and location of VA-ECMO implantation (within our center = 75% vs. peripheral hospital using our Mobile Unit of Mechanical Circulatory Support = 100%; p = .352). Survival to hospital discharge was significantly lower in patients receiving VA-ECMO during on-going cardiopulmonary resuscitation (42.8% vs. 88.0%; p = .026). CONCLUSIONS: VA-ECMO appears to be a feasible therapeutic option with a satisfactory survival rate and acceptable complications rate in poisonings complicated by refractory cardiogenic shock or cardiac arrest

Outcomes after extracorporeal life support for postcardiotomy cardiogenic shock

BACKGROUND AND AIM OF THE STUDY: Extracorporeal life support (ECLS) may be necessary in refractory postcardiotomy cardiogenic shock (PCS) unresponsive to optimal medical treatment. We sought to analyze the results and temporal outcomes of ECLS for PCS. METHODS: We performed an observational analysis of our prospective database. In order to analyze the temporal trends of ECLS for PCS, patients were divided into two groups according to the period of ECLS implantation: Group I from January 2007-June 2012, Group II from July 2012-December 2017. The primary endpoint was survival to hospital discharge. RESULTS: During the study period, 90 patients required ECLS for PCS (Group I n = 29, 32%; Group II n = 61, 68%). Mean age was 57.5 +/- 15.0 years with 62% of males. Preoperative characteristics were comparable over the two periods. A high proportion of patients were in NYHA class III/IV (61%) or cardiogenic shock (22%). Group II showed a significantly higher proportion of miscellaneous cardiac surgery operations (23 vs 3%, P = 0.031). Crossclamp and cardiopulmonary bypass times were significantly shorter in Group II (85.4 vs 114.2 min, P = 0.023 and 135.2 vs 184.2 min, P = 0.022, respectively). The complication rate during ECLS support was comparable between both groups. Successful weaning from ECLS could be accomplished in 45 (50%) patients (Group I = 52% vs Group II = 49%, P = 0.822) after a mean support of 6.4 days. Thirty-five (39%) patients survived to hospital discharge (Group I = 41% vs Group II = 38%, P = 0.738). CONCLUSIONS: Outcomes following ECLS remained stable over an 11-year period. ECLS may be limited in patients with severe preoperative cardiac dysfunction. Our data suggest that these patients may be better served with less invasive, percutaneous procedures

Central Versus Peripheral Arterial Pressure Monitoring in Patients Undergoing Cardiac Surgery: A Prospective Observational Study

OBJECTIVE: The aortic-to-radial arterial pressure gradient is described during and after cardiopulmonary bypass (CPB), and can lead to underestimating arterial blood pressure. The authors hypothesized that central arterial pressure monitoring would be associated with lower norepinephrine requirements than radial arterial pressure monitoring during cardiac surgery. DESIGN: An observational prospective cohort with propensity score analysis. SETTING: At a tertiary academic hospital's operating room and intensive care unit (ICU). PARTICIPANTS: A total of 286 consecutive adult patients undergoing cardiac surgery with CPB (central group: 109; radial group: 177) were enrolled and analyzed. INTERVENTIONS: To explore the hemodynamic effect of the measurement site, the authors divided the cohort into 2 groups according to a femoral/axillary (central group) or radial (radial group) site of arterial pressure monitoring. MEASUREMENT AND MAIN RESULTS: The primary outcome was the intraoperative amount of norepinephrine administered. Secondary outcomes included norepinephrine-free hours and ICU-free hours at postoperative day 2 (POD2). A logistic model with propensity score analysis was built to predict central arterial pressure monitoring use. The authors compared demographic, hemodynamic, and outcomes data before and after adjustment. Central group patients had a higher European System for Cardiac Operative Risk Evaluation. (EuroSCORE) compared to the radial group-7.9 ± 14.0 versus 3.8 ± 7.0, p \textless 0.001. After adjustment, both groups had similar patient EuroSCORE and arterial blood pressure levels. Intraoperative norepinephrine dose regimens were 0.10 ± 0.10 µg/kg/min in the central group and 0.11 ± 0.11 µg/kg/min in the radial group (p = 0.519). Norepinephrine-free hours at POD2 were 38 ± 17 hours versus 33 ± 19 hours in central and radial groups, respectively (p = 0.034). The ICU-free hours at POD2 were greater in the central group: 18 ± 13 hours versus 13 ± 13 hours, p = 0.008. Adverse events were less frequent in the central group than in the radial group-67% versus 50%, p = 0.007. CONCLUSIONS: No differences in the norepinephrine dose regimen were found according to the arterial measurement site during cardiac surgery. However, norepinephrine use and length of stay in the ICU were shorter, and adverse events were decreased when central arterial pressure monitoring was used

Molecular motors destroy microtubules and catalyze tubulin exchange within the lattice.

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Self-repair promotes microtubule rescue

International audienceThe dynamic instability of microtubules is characterized by slow growth phases stochastically interrupted by rapid depolymerizations called catastrophes. Rescue events can arrest the depolymerization and restore microtubule elongation. However, the origin of these rescue events remains unexplained. Here we show that microtubule lattice self-repair, in structurally damaged sites, is responsible for the rescue of microtubule growth. Tubulin photo-conversion in cells revealed that free tubulin dimers can incorporate along the shafts of microtubules, especially in regions where microtubules cross each other, form bundles or become bent due to mechanical constraints. These incorporation sites appeared to act as effective rescue sites ensuring microtubule rejuvenation. By securing damaged microtubule growth, the self-repair process supports a mechanosensitive growth by specifically promoting microtubule assembly in regions where they are subjected to physical constraints

Homogeneous multifocal excitation for high-throughput super-resolution imaging

Super-resolution microscopies have become an established tool in biological research. However, imaging throughput remains a main bottleneck in acquiring large datasets required for quantitative biology. Here we describe multifocal flat illumination for field-independent imaging (mfFIFI). By integrating mfFIFI into an instant structured illumination microscope (iSIM), we extend the field of view (FOV) to >100 x 100 mu m(2) while maintaining high-speed, multicolor, volumetric imaging at double the diffraction-limited resolution. We further extend the effective FOV by stitching adjacent images for fast live-cell super-resolution imaging of dozens of cells. Finally, we combine our flat-fielded iSIM with ultrastructure expansion microscopy to collect three-dimensional (3D) images of hundreds of centrioles in human cells, or thousands of purified Chlamydomonas reinhardtiic entrioles, per hour at an effective resolution of similar to 35 nm. Classification and particle averaging of these large datasets enables 3D mapping of posttranslational modifications of centriolar microtubules, revealing differences in their coverage and positioning