Optogenetic EB1 inactivation shortens metaphase spindles by disrupting cortical force-producing interactions with astral microtubules

Chromosome segregation is accomplished by the mitotic spindle, a bipolar micromachine built primarily from microtubules. Different microtubule populations contribute to spindle function: Kinetochore microtubules attach and transmit forces to chromosomes, antiparallel interpolar microtubules support spindle structure, and astral microtubules connect spindle poles to the cell cortex [1,2]. In mammalian cells, End Binding (EB) proteins associate with all growing microtubule plus ends throughout the cell cycle and serve as adaptors for a diverse group of +TIPs that control microtubule dynamics and interactions with other intracellular structures [3]. Because binding of many +TIPs to EB1 and thus microtubule-end association is switched off by mitotic phosphorylation [4-6] the mitotic function of EBs remains poorly understood. To analyze how EB1 and associated +TIPs on different spindle microtubule populations contribute to mitotic spindle dynamics, we use a light sensitive EB1 variant, π-EB1, that allows local, acute and reversible inactivation of +TIP association with growing microtubule ends in live cells [7]. We find that acute π-EB1 photoinactivation results in rapid and reversible metaphase spindle shortening and transient relaxation of tension across the central spindle. However, in contrast to interphase, π-EB1 photoinactivation does not inhibit microtubule growth in metaphase, but instead increases astral microtubule length and number. Yet, in the absence of EB1 activity astral microtubules fail to engage the cortical dynein/dynactin machinery and spindle poles move away from regions of π-EB1 photoinactivation. In conclusion, our optogenetic approach reveals mitotic EB1 functions that remain hidden in genetic experiments likely due to compensatory molecular systems regulating vertebrate spindle dynamics.

Inhibitory Effect of Nasal Intermittent Positive Pressure Ventilation on Gastroesophageal Reflux

Non-invasive intermittent positive pressure ventilation can lead to esophageal insufflations and in turn to gastric distension. The fact that the latter induces transient relaxation of the lower esophageal sphincter implies that it may increase gastroesophageal refluxes. We previously reported that nasal Pressure Support Ventilation (nPSV), contrary to nasal Neurally-Adjusted Ventilatory Assist (nNAVA), triggers active inspiratory laryngeal closure. This suggests that esophageal insufflations are more frequent in nPSV than in nNAVA. The objectives of the present study were to test the hypotheses that: i) gastroesophageal refluxes are increased during nPSV compared to both control condition and nNAVA; ii) esophageal insufflations occur more frequently during nPSV than nNAVA. Polysomnographic recordings and esophageal multichannel intraluminal impedance pHmetry were performed in nine chronically instrumented newborn lambs to study gastroesophageal refluxes, esophageal insufflations, states of alertness, laryngeal closure and respiration. Recordings were repeated without sedation in control condition, nPSV (15/4 cmH2O) and nNAVA (~ 15/4 cmH2O). The number of gastroesophageal refluxes recorded over six hours, expressed as median (interquartile range), decreased during both nPSV (1 (0, 3)) and nNAVA [1 (0, 3)] compared to control condition (5 (3, 10)), (p < 0.05). Meanwhile, the esophageal insufflation index did not differ between nPSV (40 (11, 61) h-1) and nNAVA (10 (9, 56) h-1) (p = 0.8). In conclusion, nPSV and nNAVA similarly inhibit gastroesophageal refluxes in healthy newborn lambs at pressures that do not lead to gastric distension. In addition, the occurrence of esophageal insufflations is not significantly different between nPSV and nNAVA. The strong inhibitory effect of nIPPV on gastroesophageal refluxes appears identical to that reported with nasal continuous positive airway pressure.

Inhibitory Effect of Nasal Intermittent Positive Pressure Ventilation on Gastroesophageal Reflux

Non-invasive intermittent positive pressure ventilation can lead to esophageal insufflations and in turn to gastric distension. The fact that the latter induces transient relaxation of the lower esophageal sphincter implies that it may increase gastroesophageal refluxes. We previously reported that nasal Pressure Support Ventilation (nPSV), contrary to nasal Neurally-Adjusted Ventilatory Assist (nNAVA), triggers active inspiratory laryngeal closure. This suggests that esophageal insufflations are more frequent in nPSV than in nNAVA. The objectives of the present study were to test the hypotheses that: i) gastroesophageal refluxes are increased during nPSV compared to both control condition and nNAVA; ii) esophageal insufflations occur more frequently during nPSV than nNAVA. Polysomnographic recordings and esophageal multichannel intraluminal impedance pHmetry were performed in nine chronically instrumented newborn lambs to study gastroesophageal refluxes, esophageal insufflations, states of alertness, laryngeal closure and respiration. Recordings were repeated without sedation in control condition, nPSV (15/4 cmH2O) and nNAVA (~ 15/4 cmH2O). The number of gastroesophageal refluxes recorded over six hours, expressed as median (interquartile range), decreased during both nPSV (1 (0, 3)) and nNAVA [1 (0, 3)] compared to control condition (5 (3, 10)), (p < 0.05). Meanwhile, the esophageal insufflation index did not differ between nPSV (40 (11, 61) h-1) and nNAVA (10 (9, 56) h-1) (p = 0.8). In conclusion, nPSV and nNAVA similarly inhibit gastroesophageal refluxes in healthy newborn lambs at pressures that do not lead to gastric distension. In addition, the occurrence of esophageal insufflations is not significantly different between nPSV and nNAVA. The strong inhibitory effect of nIPPV on gastroesophageal refluxes appears identical to that reported with nasal continuous positive airway pressure.

Usefulness of The Reflux Symptom Index in the Management of Laryngopharyngeal Reflux

Introduction Laryngopharyngeal Reflux (LPR) is highly prevalent in the general population and its impact on health systems is growing dramatically by the day. The contents of the stomach flowing back into the oesophagus, pharynx and larynx because of a transient relaxation of the lower oesophageal sphincter leads to a spectrum of symptoms diagnosed as LPR and Gastroesophageal Reflux Disease (GERD). The aim was to study in detail the symptoms of LPR and to ascertain if and how they hamper the routine of an individual by using the Reflux Symptom Index (RSI). Materials and Methods The Reflux Symptom Index (RSI) is a self-administered nine-item outcomes instrument for LPR. Ninety-one patients with clinically diagnosed LPR were taken up for this study and were issued the RSI (translated into the local language for better results) before and after treatment. Data was assessed at the end of 3 months and 6 months. Results The tabulated data showed significant improvement in the symptomatic index after treatment. Conclusion It can thus be concluded that RSI is still highly valid in the follow-up for patients with LPR because it can be easily administered and gives accurate results with excellent validation.

Dielectric polarization effect and transient relaxation in FAPbBr3 films before and after PMMA passivation

In organic–inorganic hybrid lead halide perovskites with a naturally arranged layered structure, the dielectric polarization effect caused by the dielectric mismatch between the organic and inorganic layers takes effect in their optical responses.

Some Aspects of Time-Reversal in Chemical Kinetics

Chemical kinetics govern the dynamics of chemical systems leading towards chemical equilibrium. There are several general properties of the dynamics of chemical reactions such as the existence of disparate time scales and the fact that most time scales are dissipative. This causes a transient relaxation to lower dimensional attracting manifolds in composition space. In this work, we discuss this behavior and investigate how a time reversal effects this behavior. For this, both macroscopic chemical systems as well as microscopic chemical systems (elementary reactions) are considered.

Transposon Reactivation in the Germline May Be Useful for Both Transposons and Their Host Genomes

Transposable elements (TEs) are long-term residents of eukaryotic genomes that make up a large portion of these genomes. They can be considered as perfectly fine members of genomes replicating with resident genes and being transmitted vertically to the next generation. However, unlike regular genes, TEs have the ability to send new copies to new sites. As such, they have been considered as parasitic members ensuring their own replication. In another view, TEs may also be considered as symbiotic sequences providing shared benefits after mutualistic interactions with their host genome. In this review, we recall the relationship between TEs and their host genome and discuss why transient relaxation of TE silencing within specific developmental windows may be useful for both.

P3510Cardiac function and Ca2+-cycling are different according to the level of cardiac-specific FKBP12.6 overexpression

Cardiac ryanodine receptors (RyR2) have a key role in excitation-contraction coupling by releasing Ca2+ from sarcoplasmic reticulum (SR). In cardiomyocytes, two FK506 binding protein (FKBP) isoforms have been shown to bind and to stabilize RyR2 opening: FKBP12 and FKBP12.6, the later having a stronger affinity for RyR2 despite its lower abundance. Cardiac-specific FKBP12.6 overexpressing mice have fewer arrhythmias induced by β-adrenergic stimulation than wild type (WT) mice, suggesting an implication of FKBP12.6 in an antiarrhythmic mechanism. Heart failure (HF) syndrome has a high incidence of arrhythmias, which may be explained by a decrease of FKBP isoform expression. The precise mechanism of the antiarrhythmic effect of FKBP12.6 overexpression remains unknown. To gain insight into this mechanism, we developed 2 transgenic mouse lines with cardiac-specific moderate- (TG1) and high- (TG2) FKBP12.6 overexpression levels. We characterized cardiac function, [Ca2+]i cycling and its response to β-adrenergic stimulation in both mouse lines. TG1 and TG2 mice developed mild and marked cardiac hypertrophy, respectively, associated with basal cardiac function increase in TG1 mice only. In stimulated cardiomyocytes, [Ca2+]i transient amplitude, measured by confocal microscopy, was higher in TG1 than in WT mice, without a significant difference in their SR Ca2+ content. The effect of β-adrenergic stimulation (50 nM isoproterenol) was attenuated in TG1 mice compared to WT mice, in association with the prevention of pro-arrhythmogenic Ca2+ release events, such as Ca2+ waves. In contrast, TG2 mice showed [Ca2+]i handling characteristics similar to HF, with slower [Ca2+]i transient relaxation. Interestingly, and contrary to HF, pro-arrhythmogenic Ca2+ release events were also reduced in TG2. These results indicate that the level of FKBP12.6 overexpression has distinct effects on cardiac function and on Ca2+-cycling and its response to β-adrenergic stimulation.

A Patient with Uncontrollable Belching: What to Do?

Aim.In this paper, we discuss the clinical significance of belching and present a clinical case with the description of the belching differential diagnosis.Main findings.Belching may be either a physiological or a pathological phenomenon. Pathological belching requires an adequate diagnostic approach in order to clarify the nature of its occurrence. Gastric belching occurs reflexively at the moment of stomach stretching by excess air trapped during swallowing, which initiates the transient relaxation of the lower esophageal sphincter. As a consequence, the swallowed air escapes from the stomach first into the esophagus and then into the throat. Supragastric belching is a phenomenon, in which the air entering the esophagus does not reach the stomach, but rapidly returns to the throat. In this case, unlike aerophagia, the air entering the esophagus is not accompanied by the act of swallowing. Supragastric belching is not a reflex process; rather, it is considered to be a manifestation of behavioural disorders. 24-hour pH-impedancemetry and high-resolution esophageal manometry are highly informative methods for detecting various types of belching (gastric and supragastric), as well as their mechanisms. We present a clinical observation of a 47-year-old patient suffering from supragastric belching and treated by speech therapy under the supervision of a speech therapist.Conclusion.24-hour pH-impedancemetry and high-resolution esophageal manometry (also in combination with impedancemetry) provide a complete differential diagnosis of belching and allow the most effective patient management strategy to be selected.

Synchronized mechanical oscillations at the cell–matrix interface in the formation of tensile tissue

The formation of uniaxial fibrous tissues with defined viscoelastic properties implies the existence of an orchestrated mechanical interaction between the cytoskeleton and the extracellular matrix. This study addresses the nature of this interaction. The hypothesis is that this mechanical interplay underpins the mechanical development of the tissue. In embryonic tendon tissue, an early event in the development of a mechanically robust tissue is the interaction of the pointed tips of extracellular collagen fibrils with the fibroblast plasma membrane to form stable interface structures (fibripositors). Here, we used a fibroblast-generated tissue that is structurally and mechanically matched to embryonic tendon to demonstrate homeostasis of cell-derived and external strain-derived tension over repeated cycles of strain and relaxation. A cell-derived oscillatory tension component is evident in this matrix construct. This oscillatory tension involves synchronization of individual cell forces across the construct and is induced in each strain cycle by transient relaxation and transient tensioning of the tissue. The cell-derived tension along with the oscillatory component is absent in the presence of blebbistatin, which disrupts actinomyosin force generation of the cell. The time period of this oscillation (60–90 s) is well-defined in each tissue sample and matches a primary viscoelastic relaxation time. We hypothesize that this mechanical oscillation of fibroblasts with plasma membrane anchored collagen fibrils is a key factor in mechanical sensing and feedback regulation in the formation of tensile tissues.