Systemic Human ILC Precursors Provide a Substrate for Tissue ILC Differentiation.

Innate lymphoid cells (ILCs) represent innate versions of T helper and cytotoxic T cells that differentiate from committed ILC precursors (ILCPs). How ILCPs give rise to mature tissue-resident ILCs remains unclear. Here, we identify circulating and tissue ILCPs in humans that fail to express the transcription factors and cytokine outputs of mature ILCs but have these signature loci in an epigenetically poised configuration. Human ILCPs robustly generate all ILC subsets in vitro and in vivo. While human ILCPs express low levels of retinoic acid receptor (RAR)-related orphan receptor C (RORC) transcripts, these cells are found in RORC-deficient patients and retain potential for EOMES+ natural killer (NK) cells, interferon gamma-positive (IFN-γ+) ILC1s, interleukin (IL)-13+ ILC2s, and for IL-22+, but not for IL-17A+ ILC3s. Our results support a model of tissue ILC differentiation ("ILC-poiesis"), whereby diverse ILC subsets are generated in situ from systemically distributed ILCPs in response to local environmental signals.

BCL11B and the NuRD complex cooperatively guard T-cell fate and inhibit OPA1-mediated mitochondrial fusion in T cells.

The nucleosome remodeling and histone deacetylase (NuRD) complex physically associates with BCL11B to regulate murine T-cell development. However, the function of NuRD complex in mature T cells remains unclear. Here, we characterize the fate and metabolism of human T cells in which key subunits of the NuRD complex or BCL11B are ablated. BCL11B and the NuRD complex bind to each other and repress natural killer (NK)-cell fate in T cells. In addition, T cells upregulate the NK cell-associated receptors and transcription factors, lyse NK-cell targets, and are reprogrammed into NK-like cells (ITNKs) upon deletion of MTA2, MBD2, CHD4, or BCL11B. ITNKs increase OPA1 expression and exhibit characteristically elongated mitochondria with augmented oxidative phosphorylation (OXPHOS) activity. OPA1-mediated elevated OXPHOS enhances cellular acetyl-CoA levels, thereby promoting the reprogramming efficiency and antitumor effects of ITNKs via regulating H3K27 acetylation at specific targets. In conclusion, our findings demonstrate that the NuRD complex and BCL11B cooperatively maintain T-cell fate directly by repressing NK cell-associated transcription and indirectly through a metabolic-epigenetic axis, providing strategies to improve the reprogramming efficiency and antitumor effects of ITNKs.

Measuring the α-particle charge radius with muonic helium-4 ions.

The energy levels of hydrogen-like atomic systems can be calculated with great precision. Starting from their quantum mechanical solution, they have been refined over the years to include the electron spin, the relativistic and quantum field effects, and tiny energy shifts related to the complex structure of the nucleus. These energy shifts caused by the nuclear structure are vastly magnified in hydrogen-like systems formed by a negative muon and a nucleus, so spectroscopy of these muonic ions can be used to investigate the nuclear structure with high precision. Here we present the measurement of two 2S-2P transitions in the muonic helium-4 ion that yields a precise determination of the root-mean-square charge radius of the α particle of 1.67824(83) femtometres. This determination from atomic spectroscopy is in excellent agreement with the value from electron scattering1, but a factor of 4.8 more precise, providing a benchmark for few-nucleon theories, lattice quantum chromodynamics and electron scattering. This agreement also constrains several beyond-standard-model theories proposed to explain the proton-radius puzzle2-5, in line with recent determinations of the proton charge radius6-9, and establishes spectroscopy of light muonic atoms and ions as a precise tool for studies of nuclear properties.

Human Monocytes Engage an Alternative Inflammasome Pathway.

Interleukin-1ß (IL-1ß) is a cytokine whose bioactivity is controlled by activation of the inflammasome. However, in response to lipopolysaccharide, human monocytes secrete IL-1ß independently of classical inflammasome stimuli. Here, we report that this constituted a species-specific response that is not observed in the murine system. Indeed, in human monocytes, lipopolysaccharide triggered an "alternative inflammasome" that relied on NLRP3-ASC-caspase-1 signaling, yet was devoid of any classical inflammasome characteristics including pyroptosome formation, pyroptosis induction, and K(+) efflux dependency. Genetic dissection of the underlying signaling pathway in a monocyte transdifferentiation system revealed that alternative inflammasome activation was propagated by TLR4-TRIF-RIPK1-FADD-CASP8 signaling upstream of NLRP3. Importantly, involvement of this signaling cascade was limited to alternative inflammasome activation and did not extend to classical NLRP3 activation. Because alternative inflammasome activation embraces both sensitivity and promiscuity of TLR4, we propose a pivotal role for this signaling cascade in TLR4-driven, IL-1ß-mediated immune responses and immunopathology in humans.

Transcription factors and 3D genome conformation in cell-fate decisions.

How cells adopt different identities has long fascinated biologists. Signal transduction in response to environmental cues results in the activation of transcription factors that determine the gene-expression program characteristic of each cell type. Technological advances in the study of 3D chromatin folding are bringing the role of genome conformation in transcriptional regulation to the fore. Characterizing this role of genome architecture has profound implications, not only for differentiation and development but also for diseases including developmental malformations and cancer. Here we review recent studies indicating that the interplay between transcription and genome conformation is a driving force for cell-fate decisions.

SEGCOND predicts putative transcriptional condensate-associated genomic regions by integrating multi-omics data.

MOTIVATION: The compartmentalization of biochemical reactions, involved in the activation of gene expression in the eukaryotic nucleus, leads to the formation of membraneless bodies through liquid-liquid phase separation. These formations, called transcriptional condensates, appear to play important roles in gene regulation as they are assembled through the association of multiple enhancer regions in 3D genomic space. To date, we are still lacking efficient computational methodologies to identify the regions responsible for the formation of such condensates, based on genomic and conformational data. RESULTS: In this work, we present SEGCOND, a computational framework aiming to highlight genomic regions involved in the formation of transcriptional condensates. SEGCOND is flexible in combining multiple genomic datasets related to enhancer activity and chromatin accessibility, to perform a genome segmentation. It then uses this segmentation for the detection of highly transcriptionally active regions of the genome. At a final step, and through the integration of Hi-C data, it identifies regions of putative transcriptional condensates (PTCs) as genomic domains where multiple enhancer elements coalesce in 3D space. SEGCOND identifies a subset of enhancer segments with increased transcriptional activity. PTCs are also found to significantly overlap highly interconnected enhancer elements and super enhancers obtained through two independent approaches. Application of SEGCOND on data from a well-defined system of B-cell to macrophage transdifferentiation leads to the identification of previously unreported genes with a likely role in the process. AVAILABILITY AND IMPLEMENTATION: Source code and details for the implementation of SEGCOND is available at https://github.com/AntonisK95/SEGCOND. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Bending of Lloyd's mirror to eliminate the period chirp in the fabrication of diffraction gratings.

We present a new technique to prevent the detrimental period chirp that appears in optical gratings fabricated by laser interference lithography (LIL). The idea is to bend the Lloyd's mirror in the lithographic setup to eliminate the period chirp already at the step of the grating's exposure. A new mathematical model was developed to describe the required bending geometry of the mirror. It is shown that this geometry can be described by multiple cross-sections of the mirror, each obtained by the solution of an implicit first-order differential equation. The proposed approach is illustrated on the basis of a concrete example. By slightly bending the Lloyd's mirror (by ≈ 3.5 mm of maximum deflection over an area of 142 mm × 215 mm) the period chirp of the exposed grating can be eliminated completely.

Compact 20-pass thin-disk multipass amplifier stable against thermal lensing effects and delivering 330 mJ pulses with M2 < 1.17.

We report on an Yb:YAG thin-disk multipass amplifier delivering 100 ns long pulses at a central wavelength of 1030 nm with an energy of 330 mJ at a repetition rate of 100 Hz. The beam quality factor at the maximum energy was measured to be M2 < 1.17. The small signal gain is 21.7, and the gain at 330 mJ was measured to be 6.9. The 20-pass amplifier is designed as a concatenation of stable resonator segments in which the beam is alternately Fourier transformed and relay-imaged back to the disk by a 4f-imaging optical scheme stage. The Fourier transform propagation makes the output beam robust against spherical phase front distortions, while the 4f-stage is used to compensate the thermal lens of the thin-disk and to reduce the footprint of the amplifier.

The HDAC7-TET2 epigenetic axis is essential during early B lymphocyte development.

Correct B cell identity at each stage of cellular differentiation during B lymphocyte development is critically dependent on a tightly controlled epigenomic landscape. We previously identified HDAC7 as an essential regulator of early B cell development and its absence leads to a drastic block at the pro-B to pre-B cell transition. More recently, we demonstrated that HDAC7 loss in pro-B-ALL in infants associates with a worse prognosis. Here we delineate the molecular mechanisms by which HDAC7 modulates early B cell development. We find that HDAC7 deficiency drives global chromatin de-condensation, histone marks deposition and deregulates other epigenetic regulators and mobile elements. Specifically, the absence of HDAC7 induces TET2 expression, which promotes DNA 5-hydroxymethylation and chromatin de-condensation. HDAC7 deficiency also results in the aberrant expression of microRNAs and LINE-1 transposable elements. These findings shed light on the mechanisms by which HDAC7 loss or misregulation may lead to B cell-based hematological malignancies.

General mathematical model for the period chirp in interference lithography.

We present a general analytical model for the calculation of the spatial distribution of the grating period, enabling the unification of all configurations of classical laser interference lithography (LIL) and scanning-beam interference lithography (SBIL) into one formalism. This is possible due to the consideration of Gaussian beams instead of point sources which allow for the accurate description of not only the laser's far-field but also its near-field. The proposed model enables the calculation of the grating period, the inclination and the slant of the grating lines on arbitrarily shaped substrates, originating from the interference of arbitrarily orientated and positioned Gaussian beams.

Simple spatially resolved period measurement of chirped pulse compression gratings.

We present an easy-to-implement and low-cost setup for the precise measurement of the period chirp of diffraction gratings offering a resolution of 15 pm and reasonable scan speeds of 2 seconds per measurement point. The principle of the measurement is illustrated on the example of two different pulse compression gratings, one fabricated by laser interference lithography (LIL) and the other by scanning beam interference lithography (SBIL). A period chirp of 0.22 pm/mm2 at a nominal period of 610 nm was measured for the grating fabricated with LIL, whereas no chirp was observed for the grating fabricated by SBIL, which had a nominal period of 586.2 nm.

Detrimental effects of period-chirped gratings in pulse compressors.

We present a comprehensive simulative and experimental investigation of how period-chirped pulse compression gratings affect the compressed pulses. A specifically developed ray-tracing tool was used for the simulative investigations. It is shown that the chirp creates a characteristic spatio-spectral error pattern, which leads to a degradation of the beam quality and an increase of the pulse duration. The experimental investigations, for which both a narrow-bandwidth continuous-wave and a pulsed laser beam were guided through a Treacy-compressor comprised of period-chirped gratings, confirm the simulation results and present methods on how to identify the chirp's characteristic error pattern in practice.

Injection-seeded high-power Yb:YAG thin-disk laser stabilized by the Pound-Drever-Hall method.

We demonstrate an injection-seeded thin-disk Yb:YAG laser at 1030 nm, stabilized by the Pound-Drever-Hall (PDH) method. We modified the PDH scheme to obtain an error signal free from Trojan locking points, which allowed robust re-locking of the laser and reliable long-term operation. The single-frequency pulses have 50 mJ energy (limited to avoid laser-induced damage) with a beam quality of M2 < 1.1 and an adjustable length of 55-110 ns. Heterodyne measurements confirmed a spectral linewidth of 3.7 MHz. The short pulse build-up time (850 ns) makes this laser suitable for laser spectroscopy of muonic hydrogen, pursued by the CREMA collaboration.

A new path to leukemia with WIT.

In this issue, Wang et al., 2015 describes that WT1 recruits TET2 to the DNA, an important feature of a new regulatory pathway linked to the development of acute myeloid leukemia (AML). This pathway consists of WT1, IDH1/2, and TET2 (WIT) genes, with exclusive mutations of the three genes inducing myeloid cell proliferation.

Theoretical investigation on the elimination of the period chirp by deliberate substrate deformations.

We present a theoretical investigation on the approach of deliberately bending the substrate during the exposure within laser interference lithography to compensate for the period chirp. It is shown that the yet undiscovered function of the surface geometry, necessary to achieve the zero-chirp case (i.e. having a perfectly constant period over the whole substrate) is determined by a first-order differential equation. As the direct analytical solution of this differential equation is difficult, a numerical approach is developed, based on the optimization of pre-defined functions towards the unknown analytical solution of the differential equation by means of a Nelder-Mead simplex algorithm. By applying this method to a concrete example, we show that an off-center placement of the substrate with respect to the point sources is advantageous both in terms of achievable period and substrate curvature and that a fourth-order polynomial can greatly satisfy the differential equation leading to a root-mean-square deviation of only 1.4 pm with respect to the targeted period of 610 nm.

Experimental analysis on CPA-free thin-disk multipass amplifiers operated in a helium-rich atmosphere.

We present an experimental investigation on the benefits of helium as an atmospheric gas in CPA-free thin-disk multipass amplifiers (TDMPAs) for the amplification to average powers exceeding 1 kW and pulse peak powers reaching 5 GW. Both the performance of the amplifier and the properties of the amplified sub-400 fs laser pulses centred at a wavelength of 1030 nm are compared for different helium concentrations in air, outlining and quantifying the benefits of a helium-rich atmosphere. The amplification of 100 µJ pulses in an atmosphere with 60% helium instead of air led to a maximum increase in efficiency from 24% to 29%. This translated into an increase of average output power and pulse energy of 34 W (i.e +19%) and 0.34 mJ (i.e. +19%) respectively. At the same time an improvement of the beam quality from M2 = 1.18 to M2 = 1.14 was achieved. For the amplification of 10 µJ pulses to over 1 kW of average power an atmosphere with 33% helium led to an improved beam pointing stability by a factor of 2. Moreover, the beam propagation factor M2 improved by 0.1, and the power stability improved by approximately 10%.

Simulation and compensation of thermal lensing in optical systems.

High-power optical systems are used in a number of industrial applications. One difficulty in designing such systems is that the beam itself is a significant source of heat, which changes the optical properties of the system. To reduce this effect, we propose a new thermal lensing compensation technique based on a detailed analysis of the optical properties of the high-power optical system. To this end, we have developed a new ray tracing simulation technique that accurately models optical propagation through inhomogeneous, anisotropic, and deformed media. This model enables the performance of systems in physically realistic situations to be evaluated efficiently. Experimental comparisons were conducted to validate the simulation. We found excellent agreement between the simulation and the measured data. We have validated the simulation technique for a single lens setup and a complex optical scanner system.

Nonlinear absorption in lithium triborate frequency converters for high-power ultrafast lasers.

We report on an analysis of the nonlinear absorption in lithium triborate (LBO) used for second and third harmonic generation of ultrashort laser pulses at average powers in the order of kW and with sub-picosecond pulse duration. Thermographic imaging of the LBO crystals together with a simple analytical model revealed the presence of nonlinear absorption in both harmonic generation processes. Subsequent processing with a numerical model considering the nonlinear mixing, the absorption, and the heat conduction was used to estimate the absorption coefficients. Average powers exceeding 100 W in the ultraviolet and 400 W in the visible spectral range were obtained while maintaining a good beam quality by avoiding excessive nonlinear absorption.

High-power quasi-CW diode-pumped 750-nm AlGaAs VECSEL emitting a peak power of 29.6†W and an average power of 8.5†W.

A peak output power of 29.6 W and an average output power of 8.5 W at a wavelength of 750 nm were demonstrated in quasi-CW multi-mode operation using an AlGaAs-based vertical external-cavity surface-emitting laser (VECSEL) diode-pumped at a wavelength of 675 nm. The comparatively low bandgap of the barrier material that was tuned to the pump-photon energy allowed a good compromise between low heat generation due to the quantum defect and strong absorptance of the pump radiation. The limitations for the average output power came mainly from insufficient heat flow from the intra-cavity heat spreader to the heat sink. These results show the potential for power scaling of diode-pumped VECSELs and the importance of effective heat removal.