Changes in Ocular Surface Characteristics after Switching from Benzalkonium Chloride-Preserved Latanoprost to Preservative-Free Tafluprost or Benzalkonium Chloride-Preserved Tafluprost

Purpose. The aim of the present study was to examine the effects of switching from Latanoprost ophthalmic solution containing a preservative to preservative-free Tafluprost ophthalmic solution or Tafluprost containing a preservative on ocular surfaces. Materials and Methods. Forty patients (40 eyes) with glaucoma (mean age: 62.0 ± 10.9 years) using Latanoprost with preservative for six months or longer were assigned either to a Tafluprost-containing-preservative group (20 eyes) or preservative-free-Tafluprost group (20 eyes). The intraocular pressure, corneal epithelial barrier function (fluorescein uptake concentration with fluorophotometer FL-500), superficial punctate keratopathy (AD classification), and tear film breakup time (TBUT) were assessed before switching and at 12 weeks after switching. Results. No significant differences in intraocular pressure were noted after switching in either group. Corneal epithelial barrier function was improved significantly after switching in both the Tafluprost-containing-preservative and the preservative-free-Tafluprost groups. There were no significant differences in AD scores after switching in the Tafluprost-containing-preservative group, but significant improvements were noted in the preservative-free-Tafluprost group. No significant differences in TBUT were noted in the Tafluprost-containing-preservative or preservative-free-Tafluprost groups after switching. Conclusion. After switching from preservative Latanoprost to Tafluprost containing-preservative or preservative-free Tafluprost, corneal epithelial barrier function was improved while the intraocular pressure reduction was retained

Secondary outflow driven by the protostar Ser-emb 15 in Serpens

We present the detection of a secondary outflow associated with a Class I source, Ser-emb 15, in the Serpens Molecular Cloud. We reveal two pairs of molecular outflows consisting of three lobes, namely primary and secondary outflows, using ALMA 12CO and SiO line observations at a resolution of 318 au. The secondary outflow is elongated approximately perpendicular to the axis of the primary outflow in the plane of the sky. We also identify two compact structures, Sources A and B, within an extended structure associated with Ser-emb 15 in the 1.3 mm continuum emission at a resolution of 40 au. The projected sizes of Sources A and B are 137 au and 60 au, respectively. Assuming a dust temperature of 20 K, we estimate the dust mass to be 0.0024 Msun for Source A and 0.00033 Msun for Source B. C18O line data imply the existence of rotational motion around the extended structure, however, cannot resolve rotational motion in Source A and/or B, due to insufficient angular and frequency resolutions. Therefore, we cannot conclude whether Ser-emb 15 is a single or binary system. Thus, either Source A or B could drive the secondary outflow. We discuss two scenarios to explain the driving mechanism of the primary and secondary outflows: the Ser-emb 15 system is (1) a binary system composed of Source A and B or (2) a single star system composed of only Source A. In either case, the system could be a suitable target for investigating the disk and/or binary formation processes in complicated environments. Detecting these outflows should contribute to understanding complex star-forming environments, which may be common in the star-formation processes.Comment: 21 pages, 10 figures, Accepted for publication in the Astrophysical Journa

Crescent-Shaped Molecular Outflow from the Intermediate-mass Protostar DK Cha Revealed by ALMA

We report on an Atacama Large Millimeter/submillimeter Array (ALMA) study of the Class I or II intermediate-mass protostar DK Cha in the Chamaeleon II region. The 12CO (J=2-1) images have an angular resolution of ~1'' (~250 au) and show high-velocity blueshifted (>70 km s-1) and redshifted (>50 km s-1) emissions which have 3000 au scale crescent-shaped structures around the protostellar disk traced in the 1.3mm continuum. Because the high-velocity components of the CO emission are associated with the protostar, we concluded that the emission traces the pole-on outflow. The blueshifted outflow lobe has a clear layered velocity gradient with a higher velocity component located on the inner side of the crescent shape, which can be explained by a model of an outflow with a higher velocity in the inner radii. Based on the directly driven outflow scenario, we estimated the driving radii from the observed outflow velocities and found that the driving region extends over two orders of magnitude. The 13CO emission traces a complex envelope structure with arc-like substructures with lengths of ~1000au. We identified the arc-like structures as streamers because they appear to be connected to a rotating infalling envelope. DK Cha is useful for understanding characteristics that are visible by looking at nearly face-on configurations of young protostellar systems, providing an alternative perspective for studying the star-formation process.Comment: Accepted for publication in ApJ. 12 pages, 5 figure

Ring Gap Structure around Class I Protostar WL 17

WL 17 is a Class I object and was considered to have a ring-hole structure. We analyzed the structure around WL 17 to investigate the detailed properties of WL 17. We used ALMA archival data, which have a higher angular resolution than previous observations. We investigated the WL 17 system with the 1.3 mm dust continuum and 12CO and C18O (J = 2-1) line emissions. The dust continuum emission showed a clear ring structure with inner and outer edges of ~11 and ~21 au, respectively. In addition, we detected an inner disk of < 5 au radius enclosing the central star within the ring, the first observation of this structure. Thus, WL 17 has a ring-gap structure, not a ring-hole structure. We did not detect any marked emission in either the gap or inner disk, indicating that there is no sign of a planet, circumplanetary disk, or binary companion. We identified the base of both blue-shifted and red-shifted outflows based on the 12CO emission, which is clearly associated with the disk around WL 17. The outflow mass ejection rate is ~3.6x10^-7 Msun yr-1 and the dynamical timescale is as short as ~ 10^4 yr. The C18O emission showed that an inhomogeneous infalling envelope, which can induce episodic mass accretion, is distributed in the region within ~1000 au from the central protostar. With these new findings, we can constrain the planet formation and dust growth scenarios in the accretion phase of star formation.Comment: 22 pages, 9 figures, Accepted for publication in the Astrophysical Journa

The Detection of Higher-Order Millimeter Hydrogen Recombination Lines in the Large Magellanic Cloud

We report the first extragalactic detection of the higher-order millimeter hydrogen recombination lines ($\Delta n>2$). The $\gamma$-, $\epsilon$-, and $\eta$-transitions have been detected toward the millimeter continuum source N105-1A in the star-forming region N105 in the Large Magellanic Cloud (LMC) with the Atacama Large Millimeter/submillimeter Array (ALMA). We use the H40$\alpha$ line, the brightest of the detected recombination lines (H40$\alpha$, H36$\beta$, H50$\beta$, H41$\gamma$, H57$\gamma$, H49$\epsilon$, H53$\eta$, and H54$\eta$), and/or the 3 mm free-free continuum emission to determine the physical parameters of N105-1A (the electron temperature, emission measure, electron density, and size) and study ionized gas kinematics. We compare the physical properties of N105-1A to a large sample of Galactic compact and ultracompact (UC) H II regions and conclude that N105-1A is similar to the most luminous ($L>10^5$ $L_{\odot}$) UC H II regions in the Galaxy. N105-1A is ionized by an O5.5 V star, it is deeply embedded in its natal molecular clump, and likely associated with a (proto)cluster. We incorporate high-resolution molecular line data including CS, SO, SO$_2$, and CH$_3$OH ($\sim$0.12 pc), and HCO$^{+}$ and CO ($\sim$0.087 pc) to explore the molecular environment of N105-1A. Based on the CO data, we find evidence for a cloud-cloud collision that likely triggered star formation in the region. We find no clear outflow signatures, but the presence of filaments and streamers indicates on-going accretion onto the clump hosting the UC H II region. Sulfur chemistry in N105-1A is consistent with the accretion shock model predictions.Comment: 51 pages, 30 figures, 2 tables (including appendices); accepted for publication in The Astrophysical Journal (ApJ

Safety and tolerability of bosutinib in patients with amyotrophic lateral sclerosis (iDReAM study) : A multicentre, open-label, dose-escalation phase 1 trial

Background: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease caused by the loss of motor neurons, and development of effective medicines is urgently required. Induced pluripotent stem cell (iPSC)-based drug repurposing identified the Src/c-Abl inhibitor bosutinib, which is approved for the treatment of chronic myelogenous leukemia (CML), as a candidate for the molecular targeted therapy of ALS. Methods: An open-label, multicentre, dose-escalation phase 1 study using a 3 + 3 design was conducted in 4 hospitals in Japan to evaluate the safety and tolerability of bosutinib in patients with ALS. Furthermore, the exploratory efficacy was evaluated using Revised ALS Functional Rating Scale (ALSFRS-R), predictive biomarkers including plasma neurofilament light chain (NFL) were explored, and single-cell RNA sequencing of iPSC-derived motor neurons was conducted. Patients, whose total ALSFRS-R scores decreased by 1–3 points during the 12 week, received escalating doses starting from 100 mg quaque die (QD) up to 400 mg QD based on dose-limiting toxicity (DLT) occurrence, and all participants who received one dose of the study drug were included in the primary analysis. This trial is registered with ClinicalTrials.gov, NCT04744532, as Induced pluripotent stem cell-based Drug Repurposing for Amyotrophic Lateral Sclerosis Medicine (iDReAM) study. Findings: Between March 29, 2019 and May 7, 2021, 20 patients were enrolled, 13 of whom received bosutinib treatment and 12 were included in the safety and efficacy analyses. No DLTs were observed up to 300 mg QD, but DLTs were observed in 3/3 patients of the 400 mg QD cohort. In all patients receiving 100 mg–400 mg, the prevalent adverse events (AEs) were gastrointestinal AEs in 12 patients (92.3%), liver function related AEs in 7 patients (53.8%), and rash in 3 patients (23.1%). The safety profile was consistent with that known for CML treatment, and ALS-specific AEs were not observed. A subset of patients (5/9 patients) was found to respond well to bosutinib treatment over the 12-week treatment period. It was found that the treatment-responsive patients could be distinguished by their lower levels of plasma NFL. Furthermore, single-cell RNA sequencing of iPSC-derived motor neurons revealed the pathogenesis related molecular signature in patients with ALS showing responsiveness to bosutinib. Interpretation: This is the first trial of a Src/c-Abl inhibitor, bosutinib, for patients with ALS. The safety and tolerability of bosutinib up to 300 mg, not 400 mg, in ALS were described, and responsiveness of patients on motor function was observed. Since this was an open-label trial within a short period with a limited number of patients, further clinical trials will be required

Safety and tolerability of bosutinib in patients with amyotrophic lateral sclerosis (iDReAM study): A multicentre, open-label, dose-escalation phase 1 trial

筋萎縮性側索硬化症（ALS）患者さんを対象とした ボスチニブ第1相試験；iDReAM試験の成果報告 （論文発表）. 京都大学プレスリリース. 2022-10-26.Phase I clinical trial of bosutinib for amyotrophic lateral sclerosis (ALS); iDReAM study. 京都大学プレスリリース. 2022-11-28.[Background] Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease caused by the loss of motor neurons, and development of effective medicines is urgently required. Induced pluripotent stem cell (iPSC)-based drug repurposing identified the Src/c-Abl inhibitor bosutinib, which is approved for the treatment of chronic myelogenous leukemia (CML), as a candidate for the molecular targeted therapy of ALS. [Methods] An open-label, multicentre, dose-escalation phase 1 study using a 3 + 3 design was conducted in 4 hospitals in Japan to evaluate the safety and tolerability of bosutinib in patients with ALS. Furthermore, the exploratory efficacy was evaluated using Revised ALS Functional Rating Scale (ALSFRS-R), predictive biomarkers including plasma neurofilament light chain (NFL) were explored, and single-cell RNA sequencing of iPSC-derived motor neurons was conducted. Patients, whose total ALSFRS-R scores decreased by 1–3 points during the 12-week, received escalating doses starting from 100 mg quaque die (QD) up to 400 mg QD based on dose-limiting toxicity (DLT) occurrence, and all participants who received one dose of the study drug were included in the primary analysis. This trial is registered with ClinicalTrials.gov, NCT04744532, as Induced pluripotent stem cell-based Drug Repurposing for Amyotrophic Lateral Sclerosis Medicine (iDReAM) study. [Findings] Between March 29, 2019 and May 7, 2021, 20 patients were enrolled, 13 of whom received bosutinib treatment and 12 were included in the safety and efficacy analyses. No DLTs were observed up to 300 mg QD, but DLTs were observed in 3/3 patients of the 400 mg QD cohort. In all patients receiving 100 mg–400 mg, the prevalent adverse events (AEs) were gastrointestinal AEs in 12 patients (92.3%), liver function related AEs in 7 patients (53.8%), and rash in 3 patients (23.1%). The safety profile was consistent with that known for CML treatment, and ALS-specific AEs were not observed. A subset of patients (5/9 patients) was found to respond well to bosutinib treatment over the 12-week treatment period. It was found that the treatment-responsive patients could be distinguished by their lower levels of plasma NFL. Furthermore, single-cell RNA sequencing of iPSC-derived motor neurons revealed the pathogenesis related molecular signature in patients with ALS showing responsiveness to bosutinib. [Interpretation] This is the first trial of a Src/c-Abl inhibitor, bosutinib, for patients with ALS. The safety and tolerability of bosutinib up to 300 mg, not 400 mg, in ALS were described, and responsiveness of patients on motor function was observed. Since this was an open-label trial within a short period with a limited number of patients, further clinical trials will be required

Quantitative Analysis of Phase Wave of Gene Expression in the Mammalian Central Circadian Clock Network

BACKGROUND: The suprachiasmatic nucleus (SCN), the master circadian clock, is a heterogeneous oscillator network, yet displays a robust synchronization dynamics. Recent single-cell bioluminescent imaging revealed temporal gradients in circadian clock gene expression in the SCN ex vivo. However, due to technical difficulty in biological approaches to elucidate the entire network structure of the SCN, characteristics of the gradient, which we refer to as phase wave, remain unknown. METHODOLOGY/PRINCIPAL FINDINGS: We implemented new approaches, i.e., quantitative analysis and model simulation to characterize the phase waves in Per2::Luciferase clock reporter gene expression of the rat SCN slice. Our quantitative study demonstrated not only a high degree of synchronization between the neurons and regular occurrence of the phase wave propagation, but also a significant amount of phase fluctuations contained in the wave. In addition, our simulations based on local coupling model suggest that the intercellular coupling strength estimated by the model simulations is significantly higher than the critical value for generating the phase waves. Model simulations also suggest that heterogeneity of the SCN neurons is one of the main factors causing the phase wave fluctuations. Furthermore, robustness of the SCN network against dynamical noise and variation of the natural frequencies inherent in these neurons was quantitatively assessed. CONCLUSIONS/SIGNIFICANCE: To our knowledge, this is the first quantitative evaluation of the phase wave and further characterization of the SCN neuronal network features generating the wave i.e., intercellular synchrony, phase fluctuation, strong local coupling, heterogeneous periodicity and robustness. Our present study provides an approach, which will lead to a comprehensive understanding of mechanistic and/or biological significance of the phase wave in the central circadian oscillatory system

The AAA-ATPase VPS4 Regulates Extracellular Secretion and Lysosomal Targeting of α-Synuclein

Many neurodegenerative diseases share a common pathological feature: the deposition of amyloid-like fibrils composed of misfolded proteins. Emerging evidence suggests that these proteins may spread from cell-to-cell and encourage the propagation of neurodegeneration in a prion-like manner. Here, we demonstrated that α-synuclein (αSYN), a principal culprit for Lewy pathology in Parkinson's disease (PD), was present in endosomal compartments and detectably secreted into the extracellular milieu. Unlike prion protein, extracellular αSYN was mainly recovered in the supernatant fraction rather than in exosome-containing pellets from the neuronal culture medium and cerebrospinal fluid. Surprisingly, impaired biogenesis of multivesicular body (MVB), an organelle from which exosomes are derived, by dominant-negative mutant vacuolar protein sorting 4 (VPS4) not only interfered with lysosomal targeting of αSYN but facilitated αSYN secretion. The hypersecretion of αSYN in VPS4-defective cells was efficiently restored by the functional disruption of recycling endosome regulator Rab11a. Furthermore, both brainstem and cortical Lewy bodies in PD were found to be immunoreactive for VPS4. Thus, VPS4, a master regulator of MVB sorting, may serve as a determinant of lysosomal targeting or extracellular secretion of αSYN and thereby contribute to the intercellular propagation of Lewy pathology in PD