Despite the fundamental importance of the redox metabolism of mitochondria under normal and pathological conditions, our knowledge regarding the transport of vitamin C across mitochondrial membranes ...remains far from complete. We report here that human HEK-293 cells express a mitochondrial low-affinity ascorbic acid transporter that molecularly corresponds to SVCT2, a member of the sodium-coupled ascorbic acid transporter family 2. The transporter SVCT1 is absent from HEK-293 cells. Confocal colocalization experiments with anti-SVCT2 and anti-organelle protein markers revealed that most of the SVCT2 immunoreactivity was associated with mitochondria, with minor colocalization at the endoplasmic reticulum and very low immunoreactivity at the plasma membrane. Immunoblotting of proteins extracted from highly purified mitochondrial fractions confirmed that SVCT2 protein was associated with mitochondria, and transport analysis revealed a sigmoidal ascorbic acid concentration curve with an apparent ascorbic acid transport Km of 0.6mM. Use of SVCT2 siRNA for silencing SVCT2 expression produced a major decrease in mitochondrial SVCT2 immunoreactivity, and immunoblotting revealed decreased SVCT2 protein expression by approximately 75%. Most importantly, the decreased protein expression was accompanied by a concomitant decrease in the mitochondrial ascorbic acid transport rate. Further studies using HEK-293 cells overexpressing SVCT2 at the plasma membrane revealed that the altered kinetic properties of mitochondrial SVCT2 are due to the ionic intracellular microenvironment (low in sodium and high in potassium), with potassium acting as a concentration-dependent inhibitor of SVCT2. We discarded the participation of two glucose transporters previously described as mitochondrial dehydroascorbic acid transporters; GLUT1 is absent from mitochondria and GLUT10 is not expressed in HEK-293 cells. Overall, our data indicate that intracellular SVCT2 is localized in mitochondria, is sensitive to an intracellular microenvironment low in sodium and high in potassium, and functions as a low-affinity ascorbic acid transporter. We propose that the mitochondrial localization of SVCT2 is a property shared across cells, tissues, and species.
•We describe a general mechanism for the mitochondrial acquisition of ascorbic acid.•Mitochondria express a low-affinity form of the ascorbic acid transporter SVCT2.•SVCT2 transport kinetics is altered by the intracellular ionic microenvironment.•Mitochondrial SVCT2 shows an apparent transport Km of 0.6mM.•Mitochondrial SVCT2 is fully functional under normal high ascorbate concentrations.
Large Interferometer For Exoplanets (LIFE) Quanz, S. P.; Ottiger, M.; Fontanet, E. ...
Astronomy and astrophysics (Berlin),
08/2022, Letnik:
664
Journal Article, Web Resource
Recenzirano
Odprti dostop
Context.
One of the long-term goals of exoplanet science is the atmospheric characterization of dozens of small exoplanets in order to understand their diversity and search for habitable worlds and ...potential biosignatures. Achieving this goal requires a space mission of sufficient scale that can spatially separate the signals from exoplanets and their host stars and thus directly scrutinize the exoplanets and their atmospheres.
Aims.
We seek to quantify the exoplanet detection performance of a space-based mid-infrared (MIR) nulling interferometer that measures the thermal emission of exoplanets. We study the impact of various parameters and compare the performance with that of large single-aperture mission concepts that detect exoplanets in reflected light.
Methods.
We have developed an instrument simulator that considers all major astrophysical noise sources and coupled it with Monte Carlo simulations of a synthetic exoplanet population around main-sequence stars within 20 pc of the Sun. This allows us to quantify the number (and types) of exoplanets that our mission concept could detect. Considering single visits only, we discuss two different scenarios for distributing 2.5 yr of an initial search phase among the stellar targets. Different apertures sizes and wavelength ranges are investigated.
Results.
An interferometer consisting of four 2 m apertures working in the 4–18.5 μ.m wavelength range with a total instrument throughput of 5% could detect up to ≈550 exoplanets with radii between 0.5 and 6
R
⊕
with an integrated
S
/
N
≥ 7. At least ≈160 of the detected exoplanets have radii ≤1.5
R
⊕
. Depending on the observing scenario, ≈25–45 rocky exoplanets (objects with radii between 0.5 and 1.5
R
⊕
) orbiting within the empirical habitable zone (eHZ) of their host stars are among the detections. With four 3.5 m apertures, the total number of detections can increase to up to ≈770, including ≈60–80 rocky eHZ planets. With four times 1 m apertures, the maximum detection yield is ≈315 exoplanets, including ≤20 rocky eHZ planets. The vast majority of small, temperate exoplanets are detected around M dwarfs. The impact of changing the wavelength range to 3–20 μm or 6–17 μm on the detection yield is negligible.
Conclusions.
A large space-based MIR nulling interferometer will be able to directly detect hundreds of small, nearby exoplanets, tens of which would be habitable world candidates. This shows that such a mission can compete with large single-aperture reflected light missions. Further increasing the number of habitable world candidates, in particular around solar-type stars, appears possible via the implementation of a multi-visit strategy during the search phase. The high median S/N of most of the detected planets will allow for first estimates of their radii and effective temperatures and will help prioritize the targets for a second mission phase to obtain high-S/N thermal emission spectra, leveraging the superior diagnostic power of the MIR regime compared to shorter wavelengths.
phytase (AppA) is widely used as an exogenous enzyme in monogastric animal feed mainly because of its ability to degrade phytic acid or its salt (phytate), a natural source of phosphorus. Currently, ...successful recombinant production of soluble AppA has been achieved by gene overexpression using both bacterial and yeast systems. However, some methods for the biomembrane immobilization of phytases (including AppA), such as surface display on yeast cells and bacterial spores, have been investigated to avoid expensive enzyme purification processes. This study explored a homologous protein production approach for displaying AppA on the cell surface of
by engineering its outer membrane (OM) for extracellular expression. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of total bacterial lysates and immunofluorescence microscopy of non-permeabilized cells revealed protein expression, whereas activity assays using whole cells or OM fractions indicated functional enzyme display, as evidenced by consistent hydrolytic rates on typical substrates (i.e., p-nitrophenyl phosphate and phytic acid). Furthermore, the in vitro results obtained using a simple method to simulate the gastrointestinal tract of poultry suggest that the whole-cell biocatalyst has potential as a feed additive. Overall, our findings support the notion that biomembrane-immobilized enzymes are reliable for the hydrolysis of poorly digestible substrates relevant to animal nutrition.
Context.
The theory of remote sensing shows that observing a planet at multiple phase angles (
α
) is a powerful strategy to characterize its atmosphere. Here, we study this observing strategy as ...applied to future disc-integrated direct imaging of exoplanets in reflected starlight.
Aims.
We analyse how the information contained in reflected-starlight spectra of exoplanets depends on the phase angle and the potential of multi-phase measurements to better constrain the atmospheric properties and the planet radius (
R
p
).
Methods.
We simulate spectra (500−900 nm) at
α
= 37°, 85°, and 123° with a spectral resolution of
R
~ 125−225 and signal-to-noise ratio (
S
∕
N
) = 10, consistent with the expected capabilities of future direct-imaging space telescopes. Assuming a H
2
-He atmosphere, we use a seven-parameter model that includes the atmospheric methane abundance (
f
CH
4
), the optical properties of a cloud layer and
R
p
. All these parameters are assumed to be unknown a priori and are explored with a Markov chain Monte Carlo retrieval method.
Results.
No single-phase observation can robustly identify whether the atmosphere has clouds or not. A single-phase observation at
α
= 123° and
S
∕
N
= 10 can constrain
R
p
with a maximum error of 35%, regardless of the cloud coverage. We find that combining small (37°) and large (123°) phase angles is a generally effective strategy to break multiple parameter degeneracies. This enables us to determine the presence or absence of a cloud and its main properties,
f
CH
4
and
R
p
, with higher confidence in all the explored scenarios. Other strategies, such as doubling
S
∕
N
to 20 for a single-phase observation or combining small (37°) and moderate (85°) phase angles, fail to achieve this. We show that the improvements in multi-phase retrievals are associated with the shape of the scattering phase function of the cloud aerosols and that the improvement is more modest for isotropically scattering aerosols. We finally discuss that misidentifying the background gas in the retrievals of super-Earth observations leads to systematic underestimation of the absorbing gas abundance.
Conclusions.
Exoplanets with wide ranges of observable phase angles should be prioritized for atmospheric characterization in reflected starlight.
Phosphatases are hydrolytic enzymes that cleave the phosphoester bond of numerous substrates containing phosphorylated residues. The typical classification divides them into acid or alkaline ...depending on the pH at which they have optimal activity. The histidine phosphatase (HP) superfamily is a large group of functionally diverse enzymes characterized by having an active-site His residue that becomes phosphorylated during catalysis. HP enzymes are relevant biomolecules due to their current and potential application in medicine and biotechnology.
, the causative agent of human amoebiasis, contains a gene (
) that encodes a putative secretory acid phosphatase (
HAPp49), exhibiting sequence similarity to histidine acid phosphatase (HAP)/phytase enzymes, i.e., branch-2 of HP superfamily. To assess whether it has the potential as a biocatalyst in removing phosphate groups from natural substrates, we studied the
HAPp49 structural and functional features using a computational-experimental approach. Although the combined outcome of computational analyses confirmed its structural similarity with HP branch-2 proteins, the experimental results showed that the recombinant enzyme (r
HAPp49) has negligible HAP/phytase activity. Nonetheless, results from supplementary activity evaluations revealed that r
HAPp49 exhibits Mg
-dependent alkaline pyrophosphatase activity. To our knowledge, this study represents the first computational-experimental characterization of
HAPp49, which offers further insights into the structure-function relationship and the basis for future research.
It is unknown whether individuals at high cardiovascular risk sustain a benefit in cardiovascular disease from increased olive oil consumption. The aim was to assess the association between total ...olive oil intake, its varieties (extra virgin and common olive oil) and the risk of cardiovascular disease and mortality in a Mediterranean population at high cardiovascular risk.
We included 7,216 men and women at high cardiovascular risk, aged 55 to 80 years, from the PREvención con DIeta MEDiterránea (PREDIMED) study, a multicenter, randomized, controlled, clinical trial. Participants were randomized to one of three interventions: Mediterranean Diets supplemented with nuts or extra-virgin olive oil, or a control low-fat diet. The present analysis was conducted as an observational prospective cohort study. The median follow-up was 4.8 years. Cardiovascular disease (stroke, myocardial infarction and cardiovascular death) and mortality were ascertained by medical records and National Death Index. Olive oil consumption was evaluated with validated food frequency questionnaires. Multivariate Cox proportional hazards and generalized estimating equations were used to assess the association between baseline and yearly repeated measurements of olive oil intake, cardiovascular disease and mortality.
During follow-up, 277 cardiovascular events and 323 deaths occurred. Participants in the highest energy-adjusted tertile of baseline total olive oil and extra-virgin olive oil consumption had 35% (HR: 0.65; 95% CI: 0.47 to 0.89) and 39% (HR: 0.61; 95% CI: 0.44 to 0.85) cardiovascular disease risk reduction, respectively, compared to the reference. Higher baseline total olive oil consumption was associated with 48% (HR: 0.52; 95% CI: 0.29 to 0.93) reduced risk of cardiovascular mortality. For each 10 g/d increase in extra-virgin olive oil consumption, cardiovascular disease and mortality risk decreased by 10% and 7%, respectively. No significant associations were found for cancer and all-cause mortality. The associations between cardiovascular events and extra virgin olive oil intake were significant in the Mediterranean diet intervention groups and not in the control group.
Olive oil consumption, specifically the extra-virgin variety, is associated with reduced risks of cardiovascular disease and mortality in individuals at high cardiovascular risk.
This study was registered at controlled-trials.com (http://www.controlled-trials.com/ISRCTN35739639). International Standard Randomized Controlled Trial Number (ISRCTN): 35739639. Registration date: 5 October 2005.
Context.
The direct imaging of exoplanets in reflected starlight will represent a major advance in the study of cold and temperate exoplanet atmospheres. Understanding how basic planet and ...atmospheric properties may affect the measured spectra is key to their interpretation.
Aims.
We have investigated the information content in reflected-starlight spectra of exoplanets. We apply our analysis to Barnard’s Star b candidate super-Earth, for which we assume a radius 0.6 times that of Neptune, an atmosphere dominated by H
2
–He, and a CH
4
volume mixing ratio of 5 × 10
−3
. The main conclusions of our study are however planet-independent.
Methods.
We set up a model of the exoplanet described by seven parameters including its radius, atmospheric methane abundance, and basic properties of a cloud layer. We generated synthetic spectra at zero phase (full disc illumination) from 500 to 900 nm and a spectral resolution
R
~ 125–225. We simulated a measured spectrum with a simplified, wavelength-independent noise model at a signal-to-noise ratio of 10. With a retrieval methodology based on Markov chain Monte Carlo sampling, we analysed which planet and atmosphere parameters can be inferred from the measured spectrum and the theoretical correlations amongst them. We considered limiting cases in which the planet radius is either known or completely unknown, and intermediate cases in which the planet radius is partly constrained.
Results.
If the planet radius is known, we can generally discriminate between cloud-free and cloudy atmospheres, and constrain the methane abundance to within two orders of magnitude. If the planet radius is unknown, new correlations between model parameters occur and the accuracy of the retrievals decreases. Without a radius determination, it is challenging to discern whether the planet has clouds, and the estimates on methane abundance degrade. However, we find the planet radius is constrained to within a factor of two for all the cases explored. Having a priori information on the planet radius, even if approximate, helps improve the retrievals.
Conclusions.
Reflected-starlight measurements will open a new avenue for characterizing long-period exoplanets, a population that remains poorly studied. For this task to be complete, direct-imaging observations should be accompanied by other techniques. We urge exoplanet detection efforts to extend the population of long-period planets with mass and radius determinations.
Bi2Se3, Bi2Te3, and Sb2Te3 are narrow bandgap semiconductors with tetradymite crystal structure (R‐3m) which have been extensively studied along with their alloys due to their promising operation as ...thermoelectric materials in the temperature range between 300 and 500 K. Studies on these layered semiconductors have increased tremendously in the last years since they have been recently predicted and demonstrated to behave as 3D topological insulators. In particular, a number of high‐pressure studies have been done in the recent years in these materials. In this work we summarize the main results of the high‐pressure studies performed in this family of semiconductors to date. In particular, we review recent results that address the main characteristics of the pressure‐induced electronic topological transition and structural phase transitions observed in this family of compounds. Future high‐pressure studies to be performed on these 3D topological insulators are also commented.
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