Polarized P-glycoprotein expression by the immortalised human brain endothelial cell line, hCMEC/D3, restricts apical-to-basolateral permeability to rhodamine 123

P-glycoprotein (P-gp) expression at the blood-brain barrier prevents unwanted blood-borne toxins and signalling molecules from entering the brain. Primary and immortalised human brain endothelial cells (BECs) represent two suitable options for studying P-gp function in vitro. The limited supply of primary human BECs and their instability over passage number makes this choice unattractive for medium/high throughput studies. The aim of this study was to further characterise the expression of P-gp by an immortalised human BEC line, hCMEC/D3, in order to evaluate their use as an in vitro human blood-brain barrier model. P-gp expression was stable over a high passage number (up to passage 38) and was polarised on the apical plasma membrane, consistent with human BECs in vivo. In addition, hCMEC/D3 cell P-gp expression was comparable, albeit slightly lower to that observed in primary isolated human BECs although P-gp function was similar in both cell lines. The P-gp inhibitors tariquidar and vinblastine prevented the efflux of rhodamine 123 (rh123) from hCMEC/D3 cells, indicative of functional P-gp expression. hCMEC/D3 cells also displayed polarised P-gp transport, since both tariquidar and vinblasine selectively increased the apical-to-basolateral permeability of hCMEC/D3 cells to rh123. The results presented here demonstrate that hCMEC/D3 cells are a suitable model to investigate substrate specificity of P-gp in BECs of human origin

Proteomics of industrial fungi: trends and insights for biotechnology

Filamentous fungi are widely known for their industrial applications, namely, the production of food-processing enzymes and metabolites such as antibiotics and organic acids. In the past decade, the full genome sequencing of filamentous fungi increased the potential to predict encoded proteins enormously, namely, hydrolytic enzymes or proteins involved in the biosynthesis of metabolites of interest. The integration of genome sequence information with possible phenotypes requires, however, the knowledge of all the proteins in the cell in a system-wise manner, given by proteomics. This review summarises the progress of proteomics and its importance for the study of biotechnological processes in filamentous fungi. A major step forward in proteomics was to couple protein separation with high-resolution mass spectrometry, allowing accurate protein quantification. Despite the fact that most fungal proteomic studies have been focused on proteins from mycelial extracts, many proteins are related to processes which are compartmentalised in the fungal cell, e.g. β-lactam antibiotic production in the microbody. For the study of such processes, a targeted approach is required, e.g. by organelle proteomics. Typical workflows for sample preparation in fungal organelle proteomics are discussed, including homogenisation and sub-cellular fractionation. Finally, examples are presented of fungal organelle proteomic studies, which have enlarged the knowledge on areas of interest to biotechnology, such as protein secretion, energy production or antibiotic biosynthesis

Corneal confocal microscopy detects a reduction in corneal endothelial cells and nerve fibres in patients with acute ischemic stroke

YesEndothelial dysfunction and damage underlie cerebrovascular disease and ischemic stroke. We undertook corneal confocal microscopy (CCM) to quantify corneal endothelial cell and nerve morphology in 146 patients with an acute ischemic stroke and 18 age-matched healthy control participants. Corneal endothelial cell density was lower (P<0.001) and endothelial cell area (P<0.001) and perimeter (P<0.001) were higher, whilst corneal nerve fbre density (P<0.001), corneal nerve branch density (P<0.001) and corneal nerve fbre length (P=0.001) were lower in patients with acute ischemic stroke compared to controls. Corneal endothelial cell density, cell area and cell perimeter correlated with corneal nerve fber density (P=0.033, P=0.014, P=0.011) and length (P=0.017, P=0.013, P=0.008), respectively. Multiple linear regression analysis showed a signifcant independent association between corneal endothelial cell density, area and perimeter with acute ischemic stroke and triglycerides. CCM is a rapid non-invasive ophthalmic imaging technique, which could be used to identify patients at risk of acute ischemic stroke.Qatar National Research Fund Grant BMRP2003865

Variability in Fitness Trade-Offs Amongst Coral Juveniles With Mixed Genetic Backgrounds Held in the Wild

Novel restoration methods are currently under consideration worldwide to help coral reefs recover or become more resilient to higher temperature stress. Critical field-based information concerning the paradigm of “local is best” is lacking for many methods; information which is essential to determine the risk and feasibility associated with restoration. One method involves breeding corals from different reef regions with expected variation in heat tolerance and moving those offspring to new locations to enhance offspring survival; thereby augmenting local stock to enhance survival for anticipated warming. In this study, surviving colonies from the 2016 to 2017 mass bleaching events on the Great Barrier Reef (GBR) were reproductively crossed and they included colonies sourced from northern (three) and central (two) reefs. The gravid colonies of Acropora tenuis were collected across 6° of latitude, and they were spawned to produce a total of 17 purebred and hybrid crosses. Juvenile corals (3,748 individual colonies settled on 1,474 terracotta tiles) were deployed to Davies reef in the central GBR after 4 months of aquarium rearing. Survival, growth, and coral colour (as a proxy for bleaching) were assessed after 0, 91, and 217 days of field deployment. Overall, a high percentage of juveniles (17% ± 2.5 SE) survived relative to expected survival at the final census. Survival was significantly higher for central purebred crosses, hybrid crosses had intermediate survival while northern purebreds had the lowest survival. Colour and growth rates (0.001−0.006 mm2 day–1) were not significantly different amongst central, northern, or hybrid crosses but were of a reverse pattern compared to survival. On average, northern purebred crosses grew the fastest, followed by hybrid crosses, and then central purebred crosses. Modelled growth trajectories suggest that northern purebreds would take 8 years to grow to reproductive size, hybrids would take nine, and central purebreds would require 12. All deployed juvenile corals paled over time in the field although the colour of A. tenuis juveniles did not differ significantly amongst central, northern, or hybrid crosses. Growth and survival trade-off analysis showed that although most crosses did not outperform the native central juveniles, two of the eight hybrid crosses (SBxLS, DRxCU) demonstrated faster time to reproductive age and increased survival. Overall, reduced time to reach reproductive size and minimal trade-offs in at least two of the eight hybrids suggest that these crosses may accelerate and supplement recovery through natural re-seeding of genes sourced from northern reefs

Data File 4 mitochondrial cob

Data File 4. Nexus file of aligned sequences of the mitochondrial cob gene used to generate phylogeny presented in Figure 2A

Sibling species of mutualistic Symbiodinium clade G from bioeroding sponges in the western Pacific and western Atlantic oceans

Dinoflagellates in the genus Symbiodinium associate with a broad array of metazoan and protistian hosts. Symbiodinium-based symbioses involving bioeroding sponge hosts have received less attention than those involving popular scleractinian hosts. Certain species of common Cliona harbor high densities of an ecologically restricted group of Symbiodinium, referred to as Clade G. Clade G Symbiodinium are also known to form stable and functionally important associations with Foraminifera and black corals (Antipatharia) Analyses of genetic evidence indicate that Clade G likely comprises several distinct species. Here, we use nucleotide sequence data in combination with ecological and geographic attributes to formally describe Symbiodinium endoclionum sp. nov. obtained from the Pacific boring sponge Cliona orientalis and Symbiodinium spongiolum sp. nov. from the congeneric western Atlantic sponge Cliona varians. These species appear to be part of an adaptive radiation comprising lineages of Clade G specialized to the metazoan phyla Porifera and Cnidaria, which began prior to the separation of the Pacific and Atlantic Oceans

Dosimetry and cost of imaging osseointegrated implants with film-based and computed tomography

Thermoluminescent dosimeters were used to measure radiation doses at craniofacial sites in a tissue-equivalent phantom during film-based multidirectional tomography with the Tomax Ultrascan (Incubation Industries, Ivyland, Pa.) and during computed tomography with the Elscint Excel 2400 (Elscint Corp., Tel Aviv, Israel). Mean absorbed doses for presurgical mandibular and maxillary canine and molar implant assessments were converted to equivalent doses, which were then multipied by published weighting factors and summed to give effective doses. The computed tomgraphy device consistently delivered higher doses than the Tomax Ultrascan to all anatomic locations; the differences were most pronounced when only one or two implant sites were evaluated. The reasons for the dose disparities are considered both anatomically and procedurally. A survey of examination cost revealed film-based multidirectional tomography to be less expensive than computed tomography

Data from: Sibling species of mutualistic Symbiodinium clade G from bioeroding sponges in the western Pacific and western Atlantic oceans

Dinoflagellates in the genus Symbiodinium associate with a broad array of metazoan and protistian hosts. Symbiodinium-based symbioses involving bioeroding sponge hosts have received less attention than those involving scleractinian hosts. Certain species of common Cliona harbor high densities of an ecologically restricted group of Symbiodinium, referred to as Clade G. The relationships of these unusual Clade G Symbiodinium with Foraminifera, sponges, and black coral (Antipatharia) are rarely studied. Nonetheless, analyses of genetic evidence indicate that Clade G likely comprises several distinct species. Here we use genetic data in combination with ecological and geographic evidence to formally describe Symbiodinium endoclionum sp. nov. obtained from the Pacific boring sponge Cliona orientalis and S. spongiolum sp. nov. from the congeneric western Atlantic sponge C. varians. These species appear to be part of an adaptive radiation of Clade G lineages specialized to the metazoan phyla Porifera and Cnidaria that began prior to the separation of the Pacific and Atlantic Oceans