Ryefreeman2687
Intracellular organelles change their size during trafficking and maturation. This requires the transport of ions and water across their membranes. Macropinocytosis, a ubiquitous form of endocytosis of particular importance for immune and cancer cells, generates large vacuoles that can be followed optically. Shrinkage of macrophage macropinosomes depends on TPC-mediated Na+ efflux and Cl- exit through unknown channels. Relieving osmotic pressure facilitates vesicle budding, positioning osmotic shrinkage upstream of vesicular sorting and trafficking. Here we identify the missing macrophage Cl- channel as the proton-activated Cl- channel ASOR/TMEM206. ASOR activation requires Na+-mediated depolarization and luminal acidification by redundant transporters including H+-ATPases and CLC 2Cl-/H+ exchangers. As corroborated by mathematical modelling, feedback loops requiring the steep voltage and pH dependencies of ASOR and CLCs render vacuole resolution resilient towards transporter copy numbers. TMEM206 disruption increased albumin-dependent survival of cancer cells. Our work suggests a function for the voltage and pH dependence of ASOR and CLCs, provides a comprehensive model for ion-transport-dependent vacuole maturation and reveals biological roles of ASOR.
To compare Kaplan-Meier survival curves and funnel plots for the audit of surgeon-specific corneal transplantation outcomes.
We obtained data on all patients with Fuchs endothelial dystrophy (FED) receiving a first corneal transplant in one eye between January 2012 and December 2017. We produced 2-year Kaplan-Meier graft survival curves to compare a simulated individual surgeon's graft survival rate to national pooled data. We used funnel plots to compare all surgeon outcomes to the national graft survival rate with superimposed 95 and 99.8% confidence limits. We defined an outlier as a surgeon who performed ≥10 transplants and had graft survival below the 99.8% national lower limit. To assess the effect of the surgeon case mix, we also compared unadjusted and risk-adjusted graft survival rates.
There were 3616 first corneal transplants for FED patients with complete data, performed or overseen by 196 surgeons. The 2-year national graft survival rate was 88%. The median change from the unadjusted to the risk-adjusted graft survival rate for individual surgeons was 0% (IQR 0%--2%). Of the 108 surgeons who had performed ≥10 transplants, we identified two outliers based on the unadjusted graft survival funnel plot, compared to four outliers based on the risk-adjusted graft survival funnel plot.
Funnel plots provide a visually accessible method for comparing individual graft survival rates to the national rate. Risk-adjustment accounts for clinical factors, and this has advantages for audit and clinical governance.
Funnel plots provide a visually accessible method for comparing individual graft survival rates to the national rate. Risk-adjustment accounts for clinical factors, and this has advantages for audit and clinical governance.
To investigate choroidal vascularity index (CVI) changes after oral eplerenone treatment in chronic central serous chorioretinopathy (cCSC) using the Spectral-domain (SD)-Optical Coherence Tomography (OCT) with enhanced depth imaging (EDI) mode.
Thirty-six eyes of 18 patients suffering from cCSC with monolateral foveal subretinal fluid (FSRF) successfully treated with oral eplerenone treatment and 18 age-matched healthy subjects were enroled in this retrospective study. EDI-OCT images obtained using Heidelberg Spectralis OCT device in patients with cCSC and FSRF (group 1); fellow eye (group 2) or healthy patients (healthy) were exported and then imported into image analysis ImageJ software for subsequent quantitative analysis. The main outcome measures were luminal area (LA) and CVI.
A higher value of CVI was detected in group 1 compared to healthy eyes (p = 0.006). SAR405 LA and CVI significantly reduced during follow up in group 1 and group 2. LA at 120 days was significantly lower compared to all previous time points both in group 1 and group 2 (p < 0.001). Median and [1st -3rd quartile] CVI values were 0.8 [0.7; 1.1] at baseline, 0.8 [0.7; 0.9] at 30 days; 0.7 [0.6; 0.9] at 60 and 90 days and 0.6 [0.5; 0.8] at 120 days in group 1 (p = 0.007) and 0.7 [0.6; 0.9] at baseline, 0.7 [0.7; 0.8] at 30 days; 0.7 [0.6; 0.7] at 60 and 90 days and 0.6 [0.6; 0.7] at 120 days in group 2 (p = 0.018).
Choroidal vascularity index reduced in cCSC patients after oral eplerenone treatment during follow up both in eyes with SRF and fellow eyes thus demonstrating the effectiveness of mineral corticoid receptor antagonists in recovering choroidal morphology.
Choroidal vascularity index reduced in cCSC patients after oral eplerenone treatment during follow up both in eyes with SRF and fellow eyes thus demonstrating the effectiveness of mineral corticoid receptor antagonists in recovering choroidal morphology.
Standard corneal collagen cross-linking (S-CXL) is an effective treatment to arrest Keratoconus (KC) progression in children. Less is known on the long-term efficacy of accelerated CXL (A-CXL) in paediatric populations.
A historical cohort analysis of paediatric patients (≤18 years) with KC who underwent S-CXL and A-CXL at two tertiary referral centres in Israel between 2010-2017. Preoperative and 3-year postoperative evaluation included changes in visual acuity (best spectacle corrected [BSCVA]) and uncorrected [UCVA]), refractive errors, and keratometric data.
Ninety-three eyes of 93 patients were analysed (A-CXL n = 39; S-CXL n = 54). Baseline characteristics were similar between groups. Both groups showed a significant improvement in visual acuity compared to baseline (S-CXL 0.810-0.602 LogMAR UCVA; A-CXL 0.890-0.306 LogMAR UCVA, p < 0.05 for both). Improvement in BSCVA and UCVA following A-CXL was non-inferior to S-CXL (< ± 0.2 LogMAR). Kmax decreased by a mean of 0.98 ± 5.56 dioptres following S-CXL (p = 0.02) and by 1.48 ± 8.4 dioptres following A-CXL (p = 0.015). Thinnest pachymetry decreased following both treatments (S-CXL by 26.8 ± 40.7 µm, p = 0.001, A-CXL by 10.2 ± 13.4 µm, p = 0.028), the difference between groups was within the non-inferiority margin (< ± 10 µm).
Paediatric patients followed for three years after A-CXL showed improved visual function, reduced corneal astigmatism and Kmax, and decreased thinnest corneal thickness. A-CXL was non-inferior to S-CXL at three years in terms of best-corrected and uncorrected visual acuity, thinnest pachymetry, and astigmatism. For Kmax, non-inferiority could not be concluded.
Paediatric patients followed for three years after A-CXL showed improved visual function, reduced corneal astigmatism and Kmax, and decreased thinnest corneal thickness. A-CXL was non-inferior to S-CXL at three years in terms of best-corrected and uncorrected visual acuity, thinnest pachymetry, and astigmatism. For Kmax, non-inferiority could not be concluded.
To study the outcomes of transcanalicular laser dacryocystorhinostomy (TCL-DCR) with endonasal augmentation in acute versus post-acute dacryocystitis and compare it with external DCR in post-acute settings.
A prospective, randomised study was conducted in 90 adult cases of Acute dacryocystitis. All the patients were started on systemic antibiotics and a 4 mm × 4 mm osteotomy was created using TCL-DCR. The osteotomy was enlarged to 8 mm × 8 mm by endonasal augmentation at the same sitting in group 1, after 10 days in group 2 and after 10 days with external DCR in group 3. The cases were assessed for symptomatic relief and complications. Success was defined as functional and anatomical patency at 36 months.
The mean age was 45.33 ± 15.06 years and the male female ratio was 12. The presenting complaints were painful swelling (100%), epiphora or discharge (88.8%), fistula (33%) and fever (6%). The average number of acute episodes was 2.96. The intra-group pain reduction from day 1 to day 4, was significant in all three groups (p = 0.000). Intra-operative (p = 0.015, χ
= 8.37) and post-operative complications (p = 0.002, χ
= 0.002) were higher in group. Anatomical success was achieved in all the three groups, however, the functional success in Group 3, Group 2 and Group 1 was 100%, 86.7% and 66.7% respectively (p = 0.002, χ
= 12.86).
The creation of osteotomy using TCL-DCR provides early relief in symptoms. Single-stage surgery in inflamed tissues is associated with higher complication rates. External DCR in post-acute settings gives the best outcomes with minimal complications, endoscopic augmentation requires a close follow-up.
The creation of osteotomy using TCL-DCR provides early relief in symptoms. Single-stage surgery in inflamed tissues is associated with higher complication rates. External DCR in post-acute settings gives the best outcomes with minimal complications, endoscopic augmentation requires a close follow-up.Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ORF6 is an antagonist of interferon (IFN)-mediated antiviral signaling, achieved through the prevention of STAT1 nuclear localization. However, the exact mechanism through which ORF6 prevents STAT1 nuclear trafficking remains unclear. Herein, we demonstrate that ORF6 directly binds to STAT1 with or without IFN stimulation, resulting in the nuclear exclusion of STAT1. ORF6 also recognizes importin α subtypes with different modes, in particular, high affinity to importin α1 but a low affinity to importin α5. Although ORF6 potentially disrupts the importin α/importin β1-mediated nuclear transport, thereby suppressing the nuclear translocation of the other classical nuclear localization signal-containing cargo proteins, the inhibitory effect of ORF6 is modest when compared with that of STAT1. The results indicate that the drastic nuclear exclusion of STAT1 is attributed to the specific binding with ORF6, which is a distinct strategy for the importin α1-mediated pathway. Combined with the results from a newly-produced replicon system and a hamster model, we conclude that SARS-CoV-2 ORF6 acts as a virulence factor via regulation of nucleocytoplasmic trafficking to accelerate viral replication, resulting in disease progression.Mycobacterium tuberculosis (Mtb) is responsible for approximately 1.5 million deaths each year. Though 10% of patients develop tuberculosis (TB) after infection, 90% of these infections are latent. Further, mice are nearly uniformly susceptible to Mtb but their M1-polarized macrophages (M1-MΦs) can inhibit Mtb in vitro, suggesting that M1-MΦs may be able to regulate anti-TB immunity. We sought to determine whether human MΦ heterogeneity contributes to TB immunity. Here we show that IFN-γ-programmed M1-MΦs degrade Mtb through increased expression of innate immunity regulatory genes (Inregs). In contrast, IL-4-programmed M2-polarized MΦs (M2-MΦs) are permissive for Mtb proliferation and exhibit reduced Inregs expression. M1-MΦs and M2-MΦs express pro- and anti-inflammatory cytokine-chemokines, respectively, and M1-MΦs show nitric oxide and autophagy-dependent degradation of Mtb, leading to increased antigen presentation to T cells through an ATG-RAB7-cathepsin pathway. Despite Mtb infection, M1-MΦs show increased histone acetylation at the ATG5 promoter and pro-autophagy phenotypes, while increased histone deacetylases lead to decreased autophagy in M2-MΦs.
