The alarmin interleukin-33 promotes the expansion and preserves the stemness of Tcf-1+ CD8+ T cells in chronic viral infection.

T cell factor 1 (Tcf-1) expressing CD8+ T cells exhibit stem-like self-renewing capacity, rendering them key for immune defense against chronic viral infection and cancer. Yet, the signals that promote the formation and maintenance of these stem-like CD8+ T cells (CD8+SL) remain poorly defined. Studying CD8+ T cell differentiation in mice with chronic viral infection, we identified the alarmin interleukin-33 (IL-33) as pivotal for the expansion and stem-like functioning of CD8+SL as well as for virus control. IL-33 receptor (ST2)-deficient CD8+ T cells exhibited biased end differentiation and premature loss of Tcf-1. ST2-deficient CD8+SL responses were restored by blockade of type I interferon signaling, suggesting that IL-33 balances IFN-I effects to control CD8+SL formation in chronic infection. IL-33 signals broadly augmented chromatin accessibility in CD8+SL and determined these cells' re-expansion potential. Our study identifies the IL-33-ST2 axis as an important CD8+SL-promoting pathway in the context of chronic viral infection.

Molecular and environmental determinants of biomolecular condensate formation.

Biomolecular condensate formation has been implicated in a host of biological processes and has found relevance in biology and disease. Understanding the physical principles and underlying characteristics of how these macromolecular assemblies form and are regulated has become a central focus of the field. In this Review, we introduce features of phase-separating biomolecules from a general physical viewpoint and highlight how molecular features, including affinity, valence and a competition between inter- and intramolecular contacts, affect phase separation. We then discuss sequence properties of proteins that serve to mediate intermolecular interactions. Finally, we review how the intracellular environment can affect structural and sequence determinants of proteins and modulate physical parameters of their phase transitions. The works reviewed highlight that a complex interplay exists between structure, sequence and environmental determinants in the formation of biomolecular condensates.

Blocking interleukin-23 ameliorates neuromuscular and thymic defects in myasthenia gravis.

Acetylcholine receptor (AChR) myasthenia gravis (MG) is a chronic autoimmune disease characterized by muscle weakness. The AChR+ autoantibodies are produced by B-cells located in thymic ectopic germinal centers (eGC). No therapeutic approach is curative. The inflammatory IL-23/Th17 pathway is activated in the thymus as well as in the blood and the muscle, contributing to the MG pathogenic events. We aimed to study a potential new therapeutic approach that targets IL-23p19 (IL-23) in the two complementary preclinical MG models: the classical experimental MG mouse model (EAMG) based on active immunization and the humanized mouse model featuring human MG thymuses engrafted in NSG mice (NSG-MG). In both preclinical models, the anti-IL-23 treatment ameliorated MG clinical symptoms. In the EAMG, the treatment reduced IL-17 related inflammation, anti-AChR IgG2b antibody production, activated transduction pathway involved in muscle regeneration and ameliorated the signal transduction at the neuromuscular junction. In the NSG-MG model, the treatment reduced pathogenic Th17 cell population and expression of genes involved in eGC stabilization and B-cell development in human MG thymus biopsies. Altogether, these data suggest that a therapy targeting IL-23p19 may promote significant clinical ameliorations in AChR+ MG disease due to concomitant beneficial effects on the thymus and skeletal muscle defects.

Computational Design of Single-Peptide Nanocages with Nanoparticle Templating.

Protein complexes perform a diversity of functions in natural biological systems. While computational protein design has enabled the development of symmetric protein complexes with spherical shapes and hollow interiors, the individual subunits often comprise large proteins. Peptides have also been applied to self-assembly, and it is of interest to explore such short sequences as building blocks of large, designed complexes. Coiled-coil peptides are promising subunits as they have a symmetric structure that can undergo further assembly. Here, an α-helical 29-residue peptide that forms a tetrameric coiled coil was computationally designed to assemble into a spherical cage that is approximately 9 nm in diameter and presents an interior cavity. The assembly comprises 48 copies of the designed peptide sequence. The design strategy allowed breaking the side chain conformational symmetry within the peptide dimer that formed the building block (asymmetric unit) of the cage. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) techniques showed that one of the seven designed peptide candidates assembled into individual nanocages of the size and shape. The stability of assembled nanocages was found to be sensitive to the assembly pathway and final solution conditions (pH and ionic strength). The nanocages templated the growth of size-specific Au nanoparticles. The computational design serves to illustrate the possibility of designing target assemblies with pre-determined specific dimensions using short, modular coiled-coil forming peptide sequences.

Deterministic access of broadband frequency combs in microresonators using cnoidal waves in the soliton crystal limit.

We present a method to deterministically obtain broad bandwidth frequency combs in microresonators. These broadband frequency combs correspond to cnoidal waves in the limit when they can be considered soliton crystals or single solitons. The method relies on moving adiabatically through the (frequency detuning)×(pump amplitude) parameter space, while avoiding the chaotic regime. We consider in detail Si3N4 microresonators with small or intermediate dimensions and an SiO2 microresonator with large dimensions, corresponding to prior experimental work. We also discuss the impact of thermal effects on the stable regions for the cnoidal waves. Their principal effect is to increase the detuning for all the stable regions, but they also skew the stable regions, since higher pump power corresponds to higher power and hence increased temperature and detuning. The change in the detuning is smaller for single solitons than it is for soliton crystals. Without temperature effects, the stable regions for single solitons and soliton crystals almost completely overlap. When thermal effects are included, the stable region for single solitons separates from the stable regions for the soliton crystals, explaining in part the effectiveness of backwards-detuning to obtaining single solitons.

Il-23/Th17 cell pathway: A promising target to alleviate thymic inflammation maintenance in myasthenia gravis.

IL-23/Th17 pathway has been identified to sustain inflammatory condition in several autoimmune diseases and therefore being targeted in various therapeutic and effective approaches. Patients affected with autoimmune myasthenia gravis exhibit a disease effector tissue, the thymus, that harbors ectopic germinal centers that sustain production of auto-antibodies, targeting proteins located in the neuromuscular junction, cause of the organ-specific chronic autoimmune disease. The present study aims to investigate the IL-23/Th17 cell pathway in the thymic inflammatory and pathogenic events. We found that thymuses of MG patients displayed overexpression of Interleukin-17, signature cytokine of activated Th17 cells. This activation was sustained by a higher secretion of Interleukin-23 by TEC, in addition to the increased expression of cytokines involved in Th17 cell development. The overexpression of Interleukin-23 was due to a dysregulation of interferon type I pathway. Besides, Interleukin-17 secreted, and Th17 cells were localized around thymic ectopic germinal centers. These cells expressed podoplanin, a protein involved in B-cell maturation and antibody secretion. Finally, production of Interleukin-23 was also promoted by Interleukin-17 secreted itself by Th17 cells, highlighting a chronic loop of inflammation sustained by thymic cell interaction. Activation of the IL-23/Th17 pathway in the thymus of autoimmune myasthenia gravis patients creates an unstoppable loop of inflammation that may participate in ectopic germinal center maintenance. To alleviate the physio-pathological events in myasthenia gravis patients, this pathway may be considered as a new therapeutic target.

Characterizing pump line phase offset of a single-soliton Kerr comb by dual comb interferometry.

We report phase retrieval of a single-soliton Kerr comb using electric field cross-correlation implemented via dual-comb interferometry. The phase profile of the Kerr comb is acquired through the heterodyne beat between the Kerr comb and an electro-optic comb with a pre-characterized phase profile. The soliton Kerr comb has a nearly flat phase profile, and the pump line is observed to show a phase offset which depends on the pumping parameters. The experimental results are in agreement with numerical simulations.

Infinite Assembly of Folded Proteins in Evolution, Disease, and Engineering.

Mutations and changes in a protein's environment are well known for their potential to induce misfolding and aggregation, including amyloid formation. Alternatively, such perturbations can trigger new interactions that lead to the polymerization of folded proteins. In contrast to aggregation, this process does not require misfolding and, to highlight this difference, we refer to it as agglomeration. This term encompasses the amorphous assembly of folded proteins as well as the polymerization in one, two, or three dimensions. We stress the remarkable potential of symmetric homo-oligomers to agglomerate even by single surface point mutations, and we review the double-edged nature of this potential: how aberrant assemblies resulting from agglomeration can lead to disease, but also how agglomeration can serve in cellular adaptation and be exploited for the rational design of novel biomaterials.

50-GHz-spaced comb of high-dimensional frequency-bin entangled photons from an on-chip silicon nitride microresonator.

Quantum frequency combs from chip-scale integrated sources are promising candidates for scalable and robust quantum information processing (QIP). However, to use these quantum combs for frequency domain QIP, demonstration of entanglement in the frequency basis, showing that the entangled photons are in a coherent superposition of multiple frequency bins, is required. We present a verification of qubit and qutrit frequency-bin entanglement using an on-chip quantum frequency comb with 40 mode pairs, through a two-photon interference measurement that is based on electro-optic phase modulation. Our demonstrations provide an important contribution in establishing integrated optical microresonators as a source for high-dimensional frequency-bin encoded quantum computing, as well as dense quantum key distribution.

Thermophilic Ferritin 24mer Assembly and Nanoparticle Encapsulation Modulated by Interdimer Electrostatic Repulsion.

Protein cage self-assembly enables encapsulation and sequestration of small molecules, macromolecules, and nanomaterials for many applications in bionanotechnology. Notably, wild-type thermophilic ferritin from Archaeoglobus fulgidus (AfFtn) exists as a stable dimer of four-helix bundle proteins at a low ionic strength, and the protein forms a hollow assembly of 24 protomers at a high ionic strength (∼800 mM NaCl). This assembly process can also be initiated by highly charged gold nanoparticles (AuNPs) in solution, leading to encapsulation. These data suggest that salt solutions or charged AuNPs can shield unfavorable electrostatic interactions at AfFtn dimer-dimer interfaces, but specific "hot-spot" residues controlling assembly have not been identified. To investigate this further, we computationally designed three AfFtn mutants (E65R, D138K, and A127R) that introduce a single positive charge at sites along the dimer-dimer interface. These proteins exhibited different assembly kinetics and thermodynamics, which were ranked in order of increasing 24mer propensity: A127R < wild type < D138K ≪ E65R. E65R assembled into the 24mer across a wide range of ionic strengths (0-800 mM NaCl), and the dissociation temperature for the 24mer was 98 °C. X-ray crystal structure analysis of the E65R mutant identified a more compact, closed-pore cage geometry. A127R and D138K mutants exhibited wild-type ability to encapsulate and stabilize 5 nm AuNPs, whereas E65R did not encapsulate AuNPs at the same high yields. This work illustrates designed protein cages with distinct assembly and encapsulation properties.

Soliton repetition rate in a silicon-nitride microresonator.

The repetition rate of a Kerr comb composed of a single soliton in an anomalous group velocity dispersion silicon-nitride microcavity is measured as a function of pump frequency. By comparing operation in the soliton and non-soliton states, the contributions from the Raman soliton self-frequency shift (SSFS) and the thermal effects are evaluated; the SSFS is found to dominate the changes in the repetition rate, similar to silica cavities. The relationship between the changes in the repetition rate and the pump frequency detuning is found to be independent of the nonlinearity coefficient and dispersion of the cavity. Modeling of the repetition rate change by using the generalized Lugiato-Lefever equation is discussed; the Kerr shock is found to have only a minor effect on repetition rate for cavity solitons with duration down to ∼50 fs.

Direct soliton generation in microresonators.

We investigate, numerically and experimentally, the effect of thermo-optical (TO) chaos on soliton generation dynamics in microresonators. Numerical simulations that include the thermal dynamics show that the generated solitons can either survive or annihilate when the pump laser is scanned from blue to red and then stop at a fixed wavelength; the outcome is stochastic and is strongly related to the number of solitons generated. The random fluctuations of the cavity resonance occurring under TO chaos are also found to trigger delayed spontaneous soliton generation after the laser scan ends, which could enable soliton excitation with slow laser tuning speed. Stochastic soliton annihilation/survival, as well as delayed spontaneous soliton generation, is observed experimentally in a silicon-nitride microresonator.

Improving target amino acid selectivity in a permissive aminoacyl tRNA synthetase through counter-selection.

The amino acid acridon-2-ylalanine (Acd) can be a valuable probe of protein dynamics, either alone or as part of a Förster resonance energy transfer (FRET) or photo-induced electron transfer (eT) probe pair. We have previously reported the genetic incorporation of Acd by an aminoacyl tRNA synthetase (RS). However, this RS, developed from a library of permissive RSs, also incorporates N-phenyl-aminophenylalanine (Npf), a trace byproduct of one Acd synthetic route. We have performed negative selections in the presence of Npf and analyzed the selectivity of the resulting AcdRSs by in vivo protein expression and detailed kinetic analyses of the purified RSs. We find that selection conferred a ∼50-fold increase in selectivity for Acd over Npf, eliminating incorporation of Npf contaminants, and allowing one to use a high yielding Acd synthetic route for improved overall expression of Acd-containing proteins. More generally, our report also provides a cautionary tale on the use of permissive RSs, as well as a strategy for improving selectivity for the target amino acid.

Intracavity characterization of micro-comb generation in the single-soliton regime.

Soliton formation in on-chip micro-comb generation balances cavity dispersion and nonlinearity and allows coherent, low-noise comb operation. We study the intracavity waveform of an on-chip microcavity soliton in a silicon nitride microresonator configured with a drop port. Whereas combs measured at the through port are accompanied by a very strong pump line which accounts for >99% of the output power, our experiments reveal that inside the microcavity, most of the power is in the soliton. Time-domain measurements performed at the drop port provide information that directly reflects the intracavity field. Data confirm a train of bright, close to bandwidth-limited pulses, accompanied by a weak continuous wave (CW) background with a small phase shift relative to the comb.

Integrated line-by-line optical pulse shaper for high-fidelity and rapidly reconfigurable RF-filtering.

We present a 32 channel indium phosphide integrated pulse shaper with 25 GHz channel spacing, where each channel is equipped with a semiconductor optical amplifier allowing for programmable line-by-line gain control with submicrosecond reconfigurability. We critically test the integrated pulse shaper by using it in comb-based RF-photonic filtering experiments where the precise gain control is leveraged to synthesize high-fidelity RF filters which we reconfigure on a microsecond time scale. Our on-chip pulse shaping demonstration is unmatched in its combination of speed, fidelity, and flexibility, and will likely open new avenues in the field of advanced broadband signal generation and processing.

Observation of Fermi-Pasta-Ulam Recurrence Induced by Breather Solitons in an Optical Microresonator.

We present, experimentally and numerically, the observation of Fermi-Pasta-Ulam recurrence induced by breather solitons in a high-Q SiN microresonator. Breather solitons can be excited by increasing the pump power at a relatively small pump phase detuning in microresonators. Out of phase power evolution is observed for groups of comb lines around the center of the spectrum compared to groups of lines in the spectral wings. The evolution of the power spectrum is not symmetric with respect to the spectrum center. Numerical simulations based on the generalized Lugiato-Lefever equation are in good agreement with the experimental results and unveil the role of stimulated Raman scattering in the symmetry breaking of the power spectrum evolution. Our results show that optical microresonators can be exploited as a powerful platform for the exploration of soliton dynamics.

Deterministic single soliton generation and compression in microring resonators avoiding the chaotic region.

A path within the parameter space of detuning and pump power is demonstrated in order to obtain a single cavity soliton (CS) with certainty in SiN microring resonators in the anomalous dispersion regime. Once the single CS state is reached, it is possible to continue a path to compress it, broadening the corresponding single free spectral range (FSR) Kerr frequency comb. The first step to achieve this goal is to identify the stable regions in the parameter space via numerical simulations of the Lugiato-Lefever equation (LLE). Later, using this identification, we define a path from the stable modulation instability (SMI) region to the stable cavity solitons (SCS) region avoiding the chaotic and unstable regions.

Tunable delay control of entangled photons based on dispersion cancellation.

We propose and demonstrate a novel approach for controlling the temporal position of the biphoton correlation function using pump frequency tuning and dispersion cancellation; precise waveguide engineering enables biphoton generation at different pump frequencies while the idea of nonlocal dispersion cancellation is used to create the relative signal-idler delay and simultaneously prevents broadening of their correlation. Experimental results for delay shifts up to ±15 times the correlation width are shown along with discussions of the performance metrics of this approach.

Comparative study of the oral absorption of microencapsulated ferric saccharate and ferrous sulfate in humans.

PURPOSE: Our objective was to compare the absorption of microencapsulated ferric saccharate (MFS) and ferrous sulfate (FS) in a fortified milk product, using a crossover design. METHODS: Seventeen non-iron-deficient healthy adults from both sexes participated in the study. On each intervention day (days 1 and 8), after an overnight fast, the volunteers consumed one type of product (test or control) and blood sampling was carried out at different times. The interventions days were separated by 7-day washout periods. This study was double blinded, crossover and randomized for nature of the test meals. The primary outcomes of the study were total serum iron and transferrin saturation. RESULTS: No significant differences could be observed in serum iron concentration during the 6-h postprandial study due to the type of milk product consumed, and there was neither an effect of time nor an interaction between the type of milk product and time. Transferrin saturation significantly increased after the intake of both products (P < 0.005), reaching a peak value between hours 2 and 4. No significant differences were detected between MFS and FS, indicating that iron absorption from MFS is equivalent to absorption from FS. CONCLUSIONS: MFS is a new ingredient that allows the fortification of a wide range of food products, including heat-processed and non-acidic products with similar absorption to FS, designed to produce neither organoleptic changes nor off-color development during storage of fortified food.

Prostaglandins as Candidate Ligands for a Per-ARNT-Sim (PAS) Domain of Steroid Receptor Coactivator 1 (SRC1).

Steroid receptor coactivators (SRCs) comprise a family of three paralogous proteins commonly recruited by eukaryotic transcription factors. Each SRC harbors two tandem Per-ARNT-Sim (PAS) domains that are broadly distributed that bind small molecules and regulate interactions. Using computational docking, solution NMR, mass spectrometry, and molecular dynamics simulations, we show that the SRC1 PAS-B domain can bind to certain prostaglandins (PGs) either non-covalently to a surface that overlaps with the site used to engage transcription factors or covalently to a single, specific, conserved cysteine residue next to a solvent accessible hydrophobic pocket. This pocket is in proximity to the canonical transcription factor binding site, but on the opposite side of the domain, suggesting a potential mode of regulating transcriptional activator-coactivator interactions.