Canonical α/β T-cell receptors (TCRs) bind to human leukocyte antigen (HLA) displaying antigenic peptides to elicit T cell-mediated cytotoxicity. TCR-engineered T-cell immunotherapies targeting ...cancer-specific peptide-HLA complexes (pHLA) are generating exciting clinical responses, but owing to HLA restriction they are only able to target a subset of antigen-positive patients. More recently, evidence has been published indicating that naturally occurring α/β TCRs can target cell surface proteins other than pHLA, which would address the challenges of HLA restriction. In this proof-of-concept study, we sought to identify and engineer so-called HLA-independent TCRs (HiTs) against the tumor-associated antigen mesothelin.
Using phage display, we identified a HiT that bound well to mesothelin, which when expressed in primary T cells, caused activation and cytotoxicity. We subsequently engineered this HiT to modulate the T-cell response to varying levels of mesothelin on the cell surface.
The isolated HiT shows cytotoxic activity and demonstrates killing of both mesothelin-expressing cell lines and patient-derived xenograft models. Additionally, we demonstrated that HiT-transduced T cells do not require CD4 or CD8 co-receptors and, unlike a TCR fusion construct, are not inhibited by soluble mesothelin. Finally, we showed that HiT-transduced T cells are highly efficacious in vivo, completely eradicating xenografted human solid tumors.
HiTs can be isolated from fully human TCR-displaying phage libraries against cell surface-expressed antigens. HiTs are able to fully activate primary T cells both in vivo and in vitro. HiTs may enable the efficacy seen with pHLA-targeting TCRs in solid tumors to be translated to cell surface antigens.
High-quality stoichiometric Co2Al–NO3 and Co2Al–CO3 layered double hydroxides (LDHs) have been obtained by precipitation followed by anion exchange, both high-power-sonication assisted. Application ...of high-power ultrasound has been demonstrated to result in a considerable acceleration of the crystallization process and the anion-exchange reaction. Two independent approaches were used to form bulk and 2-D samples of Co2Al–NO3 with the oriented crystallites, namely uniaxial pressing of deposits from sonicated LDH slurries and magnetic field-assisted arrangement of LDH crystallites precipitating on glass substrates. A convenient way of preparation of semi-transparent compacts with relatively big blocks of oriented crystallites have been demonstrated. Thin dense transparent films of highly-ordered crystallites of Co2Al–NO3 LDH have been produced and characterized.
The imbalance between reactive oxygen species (ROS) production and their elimination by antioxidants leads to oxidative stress. Depending on their concentration, ROS can trigger apoptosis or ...stimulate cell proliferation. We hypothesized that oxidative stress and mitochondrial dysfunction may participate not only in apoptosis detected in some myelodysplastic syndrome (MDS) patients, but also in increasing proliferation in other patients. We investigated the involvement of oxidative stress and mitochondrial dysfunction in MDS pathogenesis, as well as assessed their diagnostic and prognostic values. Intracellular peroxides, superoxide, superoxide/peroxides ratio, reduced glutathione (GSH), and mitochondrial membrane potential (Δψ
mit
) levels were analyzed in bone marrow cells from 27 MDS patients and 12 controls, by flow cytometry. We observed that all bone marrow cell types from MDS patients had increased intracellular peroxide levels and decreased GSH content, compared with control cells. Moreover, oxidative stress levels were MDS subtype- and risk group-dependent. Low-risk patients had the highest ROS levels, which can be related with their high apoptosis; and intermediate-2-risk patients had high Δψ
mit
that may be associated with their proliferative potential. GSH levels were negatively correlated with transfusion dependency, and peroxide levels were positively correlated with serum ferritin level. GSH content proved to be an accurate parameter to discriminate patients from controls. Finally, patients with high ROS or low GSH levels, as well as high superoxide/peroxides ratio had lower overall survival. Our results suggest that oxidative stress and mitochondrial dysfunction are involved in MDS development, and that oxidative stress parameters may constitute novel diagnosis and/or prognosis biomarkers for MDS.
In atherosclerotic lesions, vascular smooth muscle cells (VSMCs) represent half of the foam cell population, which is characterized by an aberrant accumulation of undigested lipids within lysosomes. ...Loss of lysosome function impacts VSMC homeostasis and disease progression. Understanding the molecular mechanisms underlying lysosome dysfunction in these cells is, therefore, crucial. We identify cholesteryl hemiazelate (ChA), a stable oxidation end-product of cholesteryl-polyunsaturated fatty acid esters, as an inducer of lysosome malfunction in VSMCs. ChA-treated VSMCs acquire a foam-cell-like phenotype, characterized by enlarged lysosomes full of ChA and neutral lipids. The lysosomes are perinuclear and exhibit degradative capacity and cargo exit defects. Lysosome luminal pH is also altered. Even though the transcriptional response machinery and autophagy are not activated by ChA, the addition of recombinant lysosomal acid lipase (LAL) is able to rescue lysosome dysfunction. ChA significantly affects VSMC proliferation and migration, impacting atherosclerosis. In summary, this work shows that ChA is sufficient to induce lysosomal dysfunction in VSMCs, that, in ChA-treated VSMCs, neither lysosome biogenesis nor autophagy are triggered, and, finally, that recombinant LAL can be a therapeutic approach for lysosomal dysfunction.
Fire is a significant agent of landscape transformation on Earth, and a dynamic and ephemeral process that is challenging to map. Difficulties include the seasonality of native vegetation in areas ...affected by fire, the high levels of spectral heterogeneity due to the spatial and temporal variability of the burned areas, distinct persistence of the fire signal, increase in cloud and smoke cover surrounding burned areas, and difficulty in detecting understory fire signals. To produce a large-scale time-series of burned area, a robust number of observations and a more efficient sampling strategy is needed. In order to overcome these challenges, we used a novel strategy based on a machine-learning algorithm to map monthly burned areas from 1985 to 2020 using Landsat-based annual quality mosaics retrieved from minimum NBR values. The annual mosaics integrated year-round observations of burned and unburned spectral data (i.e., RED, NIR, SWIR-1, and SWIR-2), and used them to train a Deep Neural Network model, which resulted in annual maps of areas burned by land use type for all six Brazilian biomes. The annual dataset was used to retrieve the frequency of the burned area, while the date on which the minimum NBR was captured in a year, was used to reconstruct 36 years of monthly burned area. Results of this effort indicated that 19.6% (1.6 million km2) of the Brazilian territory was burned from 1985 to 2020, with 61% of this area burned at least once. Most of the burning (83%) occurred between July and October. The Amazon and Cerrado, together, accounted for 85% of the area burned at least once in Brazil. Native vegetation was the land cover most affected by fire, representing 65% of the burned area, while the remaining 35% burned in areas dominated by anthropogenic land uses, mainly pasture. This novel dataset is crucial for understanding the spatial and long-term temporal dynamics of fire regimes that are fundamental for designing appropriate public policies for reducing and controlling fires in Brazil.
We identify and explore a high orbital angular momentum (OAM) harmonics generation and amplification mechanism that manipulates the OAM independently of any other laser property, by preserving the ...initial laser wavelength, through stimulated Raman backscattering in a plasma. The high OAM harmonics spectra can extend at least up to the limiting value imposed by the paraxial approximation. We show with theory and particle-in-cell simulations that the orders of the OAM harmonics can be tuned according to a selection rule that depends on the initial OAM of the interacting waves. We illustrate the high OAM harmonics generation in a plasma using several examples including the generation of prime OAM harmonics. The process can also be realized in any nonlinear optical Kerr media supporting three-wave interactions.
Dopaminergic neurotransmission in the nucleus accumbens, a central component of the mesolimbic system, has been associated with acute pain modulation. As there is a transition from acute to chronic ...pain (‘chronification’), modulatory structures may be involved in chronic pain development. Thus, this study aimed to elucidate the role of nucleus accumbens dopaminergic neurotransmission in chronification of pain. We used a rat model in which daily subcutaneous injection of prostaglandin E2 in the hindpaw for 14 days induces a long‐lasting state of nociceptor sensitization that lasts for at least 30 days following the end of the treatment. Our findings demonstrated that the increase of dopamine in the nucleus accumbens by local administration of GBR12909 (0.5 nmol/0.25 μL), a dopamine reuptake inhibitor, blocked prostaglandin E2‐induced acute hyperalgesia. This blockade was prevented by a dopamine D2 receptor antagonist (raclopride, 10 nmol/0.25 μL) but not changed by a D1 receptor antagonist (SCH23390, 0.5, 3 or 10 nmol/0.25 μL), both co‐administered with GBR12909 in the nucleus accumbens. In contrast, the induction of persistent hyperalgesia was facilitated by continuous infusion of GBR12909 in the nucleus accumbens (0.021 nmol/0.5 μL/h) over 7 days of prostaglandin E2 treatment. The development of persistent hyperalgesia was impaired by SCH23390 (0.125 nmol/0.5 μL/h) and raclopride (0.416 nmol/0.5 μL/h), both administered continuously in the nucleus accumbens over 7 days. Taken together, our data suggest that the chronification of pain involves the plasticity of dopaminergic neurotransmission in the nucleus accumbens, which switches its modulatory role from antinociceptive to pronociceptive.
Nucleus Accumbens (NAc) has been associated with pain modulation. Dopamine increase in the NAc blocks acute inflammatory hyperalgesia. This effect is mediated by dopamine D2 but not by D1 receptor. In contrast, the induction of persistent hyperalgesia is facilitated by persistent increase of dopamine in the NAc and dependent on dopamine D1 and D2 receptors. NAc dopaminergic neurotransmission is involved in the chronification of hyperalgesia and its modulatory role switches in the transition from acute to chronic condition, changing from anti to pro‐hyperalgesic.
We analyze the asymptotic behavior of the impedance (or immittance) spectroscopy response of an electrolytic cell in a finite-length situation obtained from the Poisson-Nernst-Planck (PNP) ...diffusional model and extensions by taking into account different surface effects. The analysis starts with the case characterized by perfect blocking electrodes and proceeds by considering non-blocking conditions on electrodes surface. We argue that the imaginary part of the impedance may be directly related to the boundary condition on the electrode surface, such as charge accumulation and/or transfer by electrochemical reaction or adsorption-desorption processes. We also compare the theoretical predictions with experimental data obtained for a weak electrolytic solution of KClO3.
ABSTRACT We present Chandra ACIS-S and Australia Telescope Compact Array (ATCA) radio continuum observations of the strongly lensed dusty, star-forming galaxy SPT-S J034640-5204.9 (hereafter ...SPT0346-52) at z = 5.656. This galaxy has also been observed with ALMA, HST, Spitzer, Herschel, Atacama Pathfinder EXperiment, and the Very Large Telescope. Previous observations indicate that if the infrared (IR) emission is driven by star formation, then the inferred lensing-corrected star formation rate (SFR) (∼4500 M☉ yr−1) and SFR surface density SFR (∼2000 M☉ yr−1 kpc−2) are both exceptionally high. It remained unclear from the previous data, however, whether a central active galactic nucleus (AGN) contributes appreciably to the IR luminosity. The Chandra upper limit shows that SPT0346-52 is consistent with being star formation dominated in the X-ray, and any AGN contribution to the IR emission is negligible. The ATCA radio continuum upper limits are also consistent with the FIR-to-radio correlation for star-forming galaxies with no indication of an additional AGN contribution. The observed prodigious intrinsic IR luminosity of (3.6 0.3) × 1013 L☉ originates almost solely from vigorous star formation activity. With an intrinsic source size of 0.61 0.03 kpc, SPT0346-52 is confirmed to have one of the highest SFR of any known galaxy. This high SFR, which approaches the Eddington limit for a radiation pressure supported starburst, may be explained by a combination of very high star formation efficiency and gas fraction.
The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The ...results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH● assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS●+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.
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