A Mitochondria-Dependent Pathway Mediates the Apoptosis of GSE-Induced Yeast

Grapefruit seed extract (GSE), which has powerful anti-fungal activity, can induce apoptosis in S. cerevisiae. The yeast cells underwent apoptosis as determined by testing for apoptotic markers of DNA cleavage and typical chromatin condensation by Terminal Deoxynucleotidyl Transferase–mediated dUTP Nick End Labeling (TUNEL) and 4,6′-diaminidino-2-phenylindole (DAPI) staining and electron microscopy. The changes of ΔΨmt (mitochondrial transmembrane potential) and ROS (reactive oxygen species) indicated that the mitochondria took part in the apoptotic process. Changes in this process detected by metabonomics and proteomics revealed that the yeast cells tenaciously resisted adversity. Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased. In this study, the relative changes in the levels of proteins and metabolites showed the tenacious resistance of yeast cells. However, GSE induced apoptosis in the yeast cells by destruction of the mitochondrial 60 S ribosomal protein, L14-A, and prevented the conversion of pantothenic acid to coenzyme A (CoA). The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS). We found that the changes of the metabolites and the protein changes had relevant consistency

The clinical relevance of oliguria in the critically ill patient : Analysis of a large observational database

Funding Information: Marc Leone reports receiving consulting fees from Amomed and Aguettant; lecture fees from MSD, Pfizer, Octapharma, 3 M, Aspen, Orion; travel support from LFB; and grant support from PHRC IR and his institution. JLV is the Editor-in-Chief of Critical Care. The other authors declare that they have no relevant financial interests. Publisher Copyright: © 2020 The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Background: Urine output is widely used as one of the criteria for the diagnosis and staging of acute renal failure, but few studies have specifically assessed the role of oliguria as a marker of acute renal failure or outcomes in general intensive care unit (ICU) patients. Using a large multinational database, we therefore evaluated the occurrence of oliguria (defined as a urine output 16 years) patients in the ICON audit who had a urine output measurement on the day of admission were included. To investigate the association between oliguria and mortality, we used a multilevel analysis. Results: Of the 8292 patients included, 2050 (24.7%) were oliguric during the first 24 h of admission. Patients with oliguria on admission who had at least one additional 24-h urine output recorded during their ICU stay (n = 1349) were divided into three groups: transient - oliguria resolved within 48 h after the admission day (n = 390 [28.9%]), prolonged - oliguria resolved > 48 h after the admission day (n = 141 [10.5%]), and permanent - oliguria persisting for the whole ICU stay or again present at the end of the ICU stay (n = 818 [60.6%]). ICU and hospital mortality rates were higher in patients with oliguria than in those without, except for patients with transient oliguria who had significantly lower mortality rates than non-oliguric patients. In multilevel analysis, the need for RRT was associated with a significantly higher risk of death (OR = 1.51 [95% CI 1.19-1.91], p = 0.001), but the presence of oliguria on admission was not (OR = 1.14 [95% CI 0.97-1.34], p = 0.103). Conclusions: Oliguria is common in ICU patients and may have a relatively benign nature if only transient. The duration of oliguria and need for RRT are associated with worse outcome.publishersversionPeer reviewe

Unlikely SARS-CoV-2 vertical transmission from mother to child: A case report

As the 2019 novel coronavirus disease (COVID-19) rapidly spread across China and to more than 70 countries, an increasing number of pregnant women were affected. The vertical transmission potential of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of great concern to the obstetrics, neonatologists, and public health agencies. Though some studies indicated the risk of vertical transmission is low, few cases have been reported with comprehensive serial tests from multiple specimens. In this case, a female preterm infant was born to a mother with confirmed COVID-19. She presented with mild respiratory distress and received general management and a short period of nasal continuous positive airway pressure support. During her stay at the hospital, a series of SARS-CoV-2 nucleic test from her throat and anal swab, serum, bronchoalveolar lavage fluid, and urine were negative. The nucleic acid test from the mother's amniotic fluid, vaginal secretions, cord blood, placenta, serum, anal swab, and breast milk were also negative. The most comprehensively tested case reported to date confirmed that the vertical transmission of COVID is unlikely, but still, more evidence is needed

A novel mutation in the RPE65 gene causing Leber congenital amaurosis and its transcriptional expression in vitro.

The retinal pigment epithelium-specific 65 kDa protein is an isomerase encoded by the RPE65 gene (MIM 180069) that is responsible for an essential enzymatic step required for the function of the visual cycle. Mutations in the RPE65 gene cause not only subtype II of Leber congenital amaurosis (LCA) but also early-onset severe retinal dystrophy (EOSRD). This study aims to investigate a Chinese case diagnosed as EOSRD and to characterize the polymorphisms of the RPE65 gene. A seven-year-old girl with clinical symptoms of EOSRD and her parents were recruited into this study. Ophthalmologic examinations, including best-corrected visual acuity, slit-lamp, Optical coherence tomography (OCT), and fundus examination with dilated pupils, were performed to determine the clinical characteristics of the whole family. We amplified and sequenced the entire coding region and adjacent intronic sequences of the coding regions of the RPE65 gene for the whole family to explore the possible mutation. Our results demonstrate that the patient exhibited the typical clinically features of EOSRD. Her bilateral decimal visual acuity was 0.3 and 0.4 in the left and right eyes, respectively. Spectral-domain optical coherence tomography (SD-OCT) was used to assess the retinal stratification for the whole family. All together, we identified four mutations within the RPE65 gene (c.1056G>A, c.1243+2T>A, c.1338+20A>C and c.1590C>A) in the patient. Among the four mutations, c.1056G>A and c.1338+20A>C had been reported previously and another two were found for the first time in this study. Her mother also carried the novel mutation (c.1243+2T>A). Either a single or a compound heterozygous or a homozygous one mutation is expected to cause EOSRD because mutations of RPE65 gene usually cause an autosomal recessive disease. Therefore, we speculate that the c.1590C>A mutation together with the c.1243+2T>A mutation may cause the patient's phenotype

Pedigree of the Chinese family and mutations of <i>RPE65</i> gene.

<p>In the family structure, male and female are represented by squares and circles, respectively. The filled square symbol represents the ESORD-affected daughter (c). One mutation (c.1338+20A>C) is detected in <i>RPE65</i> gene of her father (a); two other mutations (c.1243+2T>A and c.1590C>A) are detected in her mother (b); four mutation are found in the daughter (a). The red color highlights the novel mutation.</p

Fundus photographs of both eyes in the family.

<p>Color fundus photographs of both eyes (a: left eye; b: right eye) show mildly attenuated retinal vessels, some whitish dots, and numerous grayish deposits in the mid-peripheral retina of the patient. The inserted panels (g, h) show a magnification of the indicated areas. Whitish dots are marked by white arrowheads. Fundus photographs of her father (c, d) and her mother (e, f) show no whitish dots or grayish deposits in the mid-peripheral retina.</p

Structure of <i>RPE65 </i>minigene.

<p>The pCIneo minigenes of <i>RPE65</i> gene were constructed to contain three exons (exon 11, 12, and 13) and flanking intronic sequences (intron 11 and 12) from wild or mutant type (c.1243+2T>A) of <i>RPE65</i> gene. <i>Eco</i>RI and <i>Sal</i>I represent restriction enzyme sites. Horizontal arrows indicate the positions and the directions of the primers. The red 483 bp indicates the amplified product.</p

Primers used in the study.

<p>Notes: *denotes primers for amplification of the wild and mutant fragments of exons 11, 12, and 13 of <i>RPE65</i> gene. Bold and underlined sequences are restriction enzyme sites.</p><p>Primers used in the study.</p

Analysis of pre-mRNA splicing of pCIneo minigenes in the transfected 293T cell line.

<p>A. Graphic representation of pre-mRNA splicing of wild type and mutant (C.1243+2T>A) minigenes of <i>RPE65</i> gene. B. The isolated RNA of transfected cells was amplified by RT-PCR analysis. The different splicing products for wild type (264 bp) and mutant (358 bp) are shown on a 2% agarose gel. The mutant lane demonstrated that only a 358 bp band was obtained from the mutant-RPE65 minigene (a). The wild-type lane showed two different size DNA bands: one is a 264 bp band, and another is ∼200 bp band. The 264 bp DNA band is the expected size (a). The 200 bp band is the amplification caused by mispriming.</p