Mcm5 Represses Endodermal Migration through Cxcr4a-itgb1b Cascade Instead of Cell Cycle Control

Minichromosome maintenance protein 5 (MCM5) is a critical cell cycle regulator; its role in DNA replication is well known, but whether it is involved in the regulation of organogenesis in a cell cycle-independent way, is far from clear. In this study, we found that a loss of mcm5 function resulted in a mildly smaller liver, but that mcm5 overexpression led to liver bifida. Further, the data showed that mcm5 overexpression delayed endodermal migration in the ventral–dorsal axis and induced the liver bifida. Cell cycle analysis showed that a loss of mcm5 function, but not overexpression, resulted in cell cycle delay and increased cell apoptosis during gastrulation, implying that liver bifida was not the result of a cell cycle defect. In terms of its mechanism, our data proves that mcm5 represses the expression of cxcr4a, which sequentially causes a decrease in the expression of itgb1b during gastrulation. The downregulation of the cxcr4a-itgb1b cascade leads to an endodermal migration delay during gastrulation, as well as to the subsequent liver bifida during liver morphogenesis. In conclusion, our results suggest that in a cell cycle-independent way, mcm5 works as a gene expression regulator, either partially and directly, or indirectly repressing the expression of cxcr4a and the downstream gene itgb1b, to coordinate endodermal migration during gastrulation and liver location during liver organogenesis

Atrasentan and renal events in patients with type 2 diabetes and chronic kidney disease (SONAR): a double-blind, randomised, placebo-controlled trial

Background: Short-term treatment for people with type 2 diabetes using a low dose of the selective endothelin A receptor antagonist atrasentan reduces albuminuria without causing significant sodium retention. We report the long-term effects of treatment with atrasentan on major renal outcomes. Methods: We did this double-blind, randomised, placebo-controlled trial at 689 sites in 41 countries. We enrolled adults aged 18–85 years with type 2 diabetes, estimated glomerular filtration rate (eGFR)25–75 mL/min per 1·73 m 2 of body surface area, and a urine albumin-to-creatinine ratio (UACR)of 300–5000 mg/g who had received maximum labelled or tolerated renin–angiotensin system inhibition for at least 4 weeks. Participants were given atrasentan 0·75 mg orally daily during an enrichment period before random group assignment. Those with a UACR decrease of at least 30% with no substantial fluid retention during the enrichment period (responders)were included in the double-blind treatment period. Responders were randomly assigned to receive either atrasentan 0·75 mg orally daily or placebo. All patients and investigators were masked to treatment assignment. The primary endpoint was a composite of doubling of serum creatinine (sustained for ≥30 days)or end-stage kidney disease (eGFR <15 mL/min per 1·73 m 2 sustained for ≥90 days, chronic dialysis for ≥90 days, kidney transplantation, or death from kidney failure)in the intention-to-treat population of all responders. Safety was assessed in all patients who received at least one dose of their assigned study treatment. The study is registered with ClinicalTrials.gov, number NCT01858532. Findings: Between May 17, 2013, and July 13, 2017, 11 087 patients were screened; 5117 entered the enrichment period, and 4711 completed the enrichment period. Of these, 2648 patients were responders and were randomly assigned to the atrasentan group (n=1325)or placebo group (n=1323). Median follow-up was 2·2 years (IQR 1·4–2·9). 79 (6·0%)of 1325 patients in the atrasentan group and 105 (7·9%)of 1323 in the placebo group had a primary composite renal endpoint event (hazard ratio [HR]0·65 [95% CI 0·49–0·88]; p=0·0047). Fluid retention and anaemia adverse events, which have been previously attributed to endothelin receptor antagonists, were more frequent in the atrasentan group than in the placebo group. Hospital admission for heart failure occurred in 47 (3·5%)of 1325 patients in the atrasentan group and 34 (2·6%)of 1323 patients in the placebo group (HR 1·33 [95% CI 0·85–2·07]; p=0·208). 58 (4·4%)patients in the atrasentan group and 52 (3·9%)in the placebo group died (HR 1·09 [95% CI 0·75–1·59]; p=0·65). Interpretation: Atrasentan reduced the risk of renal events in patients with diabetes and chronic kidney disease who were selected to optimise efficacy and safety. These data support a potential role for selective endothelin receptor antagonists in protecting renal function in patients with type 2 diabetes at high risk of developing end-stage kidney disease. Funding: AbbVie

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049

Effects of different concentrations of dimethylsulfoxide on development of zebrafish embryos and dopamine neurons

bjective To study the effect of exposures to different concentrations of dimethyl sulfoxide (DMSO) on the development of zebrafish embryos and dopaminergic (DA) neurons. Methods Tg(Vmat2 :GFP) zebrafish embryos of the same generation were randomized into 10 groups at 6 h post-fertilization for exposures to 9 concentrations (0.1%, 0.3%, 0.5%, 0.8%, 1.0%, 1.5%, 2.0%, 2.5%, and 3.0%; V/V) of DMSO for 72 h or without any exposure (blank control group). The survival rate, deformity rate, hatching rate and behavioral changes of the embryos were analyzed. The development of DA neurons was observed using laser confocal microscopy, and the cell proliferation and apoptosis were detected using PH3 antibody and an apoptosis detection kit, respectively. Results Exposure to 2.5% DMSO significantly decreased the survival rate of the zebrafish embryos (P < 0.01); at the concentration of 2.0%, DMSO exposure caused an obvious increase in the malformation rate (P < 0.01) and a decrease in the hatching rate of the embryos (P < 0.05); DMSO at 2.5% significantly decreased the frequency of tail swing of the zebrafish embryos (P < 0.05). Zebrafish embryos exposed to 0.5% DMSO showed a significantly reduced number of DA neurons (P < 0.05). Exposure of the zebrafish embryos to DMSO at a concentration above 0.3% significantly reduced the proliferation of the diencephalon cells (P < 0.01), and a DMSO concentration above 0.8% could obviously increase apoptosis of the dopaminergic neurons. Conclusion DMSO exposure causes developmental disorders in zebrafish embryos, and its neurotoxicity can result in loss of dopaminergic neurons in the diencephalon and suppress the cell proliferation

Prediction of sinus node dysfunction in patients with paroxysmal atrial fibrillation and sinus pause

Background and Objectives: Sinus pause association of paroxysmal atrial fibrillation (PAP) is generally considered a sinus node dysfunction (SND) known as tachycardia-bradycardia syndrome (TBS). Cure of tachyarrhythmias in some patients with prolonged pauses on termination of tachyarrhythmia resulted in an improvement in sinus node function and/or a return to normal ranges thus avoiding the need for permanent pacing. The purpose of this study was to investigate the electrophysiological and clinical characteristics as well as their sinus node function in patients with prolonged pauses and PAP. Methods and Results: Of 1266 PAP patients undergoing radiofrequency catheter ablation (RFCA), 122 patients with pauses (>2 s) were studied. The mean maximum symptomatic prolonged pause on termination of tachycardias was 2.67 ± 1.4 s (2.0–12.6 s). SND developed in 32 patients. There was no difference between the patients with and without SND in terms of the age and sex. Patients were divided into two groups based on the intracardiac electrophysiology study after the RFCA procedure. Eleven patients (8 men, 3 women; age 30–72 years, mean 48.2 ± 9.3 years) were placed in a normal sinus node function (NSF) group, and 17 patients (12 men, 5 women; age 37–75 years, mean 62.2 ± 7.5 years) were in an SND group. There was no significant difference in gender, left atrial dimension, left ventricular ejection fraction, blood pressure, or the longest pause between the two groups. RFCA was successful in 28 patients. Electrophysiological data show that corrected sinus node recovery time (CSNRT) was 409 ± 152 ms in NSF group patients and 558 ± 178 ms in SND group before the ablation procedure (P 2 cycle lengths in 12 patients. In the multivariate analysis, prolonged pauses on termination of tachyarrhythmia, frequency of pauses, and mean heart rate after ablation were independent predictors of SND. Conclusions: (1) Catheter ablation of atrial fibrillation (AF) is effective in treating paroxysmal AF-related TBS; (2) in some patients, there is progressive improvement of sinus node function after elimination of AF, an indication that these sinus pauses may be a manifestation of tachycardia-mediated remodeling of the sinus node, and that permanent pacemaker implantation is unnecessary; (3) among the extrinsic and intrinsic causes of SND, intrinsic causes are seldom reversible and some causes of extrinsic SND may be reversible; and (4) prolonged pauses on termination of PAF, frequency of pauses (>2.0 s), and mean heart rate after ablation were independent predictive factors of SND

Spatial and Temporal Distribution of Dopaminergic Neurons during Development in Zebrafish

As one of the model organisms of Parkinson’s disease (PD) research, the zebrafish has its advantages, such as the 87% homology with human genome and transparent embryos which make it possible to observe the development of dopaminergic neurons in real time. However, there is no midbrain dopaminergic system in zebrafish when compared with mammals, and the location and projection of the dopaminergic neurons is seldom reported. In this study, Vmat2:GFP transgenic zebrafish was used to observe the development and distribution of dopaminergic neurons in real time. We found that diencephalon (DC) 2 and DC4 neuronal populations were detected at 24 hpf. All DC neuronal populations as well as those in locus coeruleus, raphe nuclei and telencephalon were detected at 48 hpf. Axons were detected at 72 hpf. At 96 hpf, all the neuronal populations were detected. For the first time we reported axons from the posterior tubercle of ventral diencephalon projected to subpallium in vivo. However, when compared with results from whole mount tyrosine hydroxylase immunofluorescence staining in wild type zebrafish, we found that DC2 and DC4 neuronal populations were mainly dopaminergic, while DC1, DC3, DC5 and DC6 might not. Neurons in pretectum and telencephalon were mainly dopaminergic, while neurons in locus coeruleus and raphe nuclei might be noradrenergic. Our study makes some corrections and modifications on the development, localization and distribution of zebrafish dopaminergic neurons, and provides some experimental evidences for the construction of the zebrafish PD model

JOURNAL OF NEUROINFLAMMATION CORRECTION Open Access

adapter-inducing interferon-β (TRIF) deficiency promotes retinal ganglion cell survival and axon regeneration via nuclear factor-κ