Cellulose degradation by polysaccharide monooxygenases.

Polysaccharide monooxygenases (PMOs), also known as lytic PMOs (LPMOs), enhance the depolymerization of recalcitrant polysaccharides by hydrolytic enzymes and are found in the majority of cellulolytic fungi and actinomycete bacteria. For more than a decade, PMOs were incorrectly annotated as family 61 glycoside hydrolases (GH61s) or family 33 carbohydrate-binding modules (CBM33s). PMOs have an unusual surface-exposed active site with a tightly bound Cu(II) ion that catalyzes the regioselective hydroxylation of crystalline cellulose, leading to glycosidic bond cleavage. The genomes of some cellulolytic fungi contain more than 20 genes encoding cellulose-active PMOs, suggesting a diversity of biological activities. PMOs show great promise in reducing the cost of conversion of lignocellulosic biomass to fermentable sugars; however, many questions remain about their reaction mechanism and biological function. This review addresses, in depth, the structural and mechanistic aspects of oxidative depolymerization of cellulose by PMOs and considers their biological function and phylogenetic diversity.

Scan-less 3D microscopy based on spatiotemporal encoding on a single-cavity dual-comb laser.

Dual-comb microscopy enables high-speed and high-precision optical sampling by simultaneously extracting both amplitude and phase information from the interference signals with frequency division multiplexing. In this Letter, we introduce a spatiotemporal encoding approach for dual-comb microscopy that overcomes previous limitations such as mechanical scanning, low sampling efficiency, and system complexity. By employing free-space angular-chirp-enhanced delay (FACED) and a low-noise single-cavity dual-comb laser, we achieve scan-less 3D imaging with nanometer precision and a 3D distance-imaging rate of 330 Hz, restricted only by the repetition rate difference of the dual-comb laser. Specifically, the FACED unit linearly arranges the laser beam into an array. A grating subsequently disperses this array transversely into lines, facilitating ultrafast spectroscopic applications that are 1-2 orders of magnitude quicker than traditional dual-comb methods. This spatiotemporal encoding also eases the stringent conditions on various dual-comb laser parameters, such as repetition rates, coherence, and stability. Through carefully designed experiments, we demonstrate that our scan-less system can measure 3D profiles of microfabricated structures at a rate of 7 million pixels per second. Our method significantly enhances measurement speed while maintaining high precision, using a compact light source. This advancement has the potential for broad applications, including phase imaging, surface topography, distance ranging, and spectroscopy.

THz-TDS with gigahertz Yb-based dual-comb lasers: noise analysis and mitigation strategies.

We investigate terahertz time-domain spectroscopy using a low-noise dual-frequency-comb laser based on a single spatially multiplexed laser cavity. The laser cavity includes a reflective biprism, which enables generation of a pair of modelocked output pulse trains with slightly different repetition rates and highly correlated noise characteristics. These two pulse trains are used to generate the THz waves and detect them by equivalent time sampling. The laser is based on Yb:CALGO, operates at a nominal repetition rate of 1.18 GHz, and produces 110 mW per comb with 77 fs pulses around 1057 nm. We perform THz measurements with Fe-doped photoconductive antennas, operating these devices with gigahertz 1 µm lasers for the first time, to our knowledge, and obtain THz signal currents approximately as strong as those from reference measurements at 1.55 µm and 80 MHz. We investigate the influence of the laser's timing noise properties on THz measurements, showing that the laser's timing jitter is quantitatively explained by power-dependent shifts in center wavelength. We demonstrate reduction in noise by simple stabilization of the pump power and show up to 20 dB suppression in noise by the combination of shared pumping and shared cavity architecture. The laser's ultra-low-noise properties enable averaging of the THz waveform for repetition rate differences from 1 kHz to 22 kHz, resulting in a dynamic range of 55 dB when operating at 1 kHz and averaging for 2 s. We show that the obtained dynamic range is competitive and can be well explained by accounting for the measured optical delay range, integration time, as well as the measurement bandwidth dependence of the noise from transimpedance amplification. These results will help enable a new approach to high-resolution THz-TDS enabled by low-noise gigahertz dual-comb lasers.

Ultrafast Yb:YAG laser oscillator with gigahertz repetition rate.

We present a SESAM modelocked Yb:YAG solid-state laser providing low-noise narrowband pulses with a pulse duration of 606 fs at a 1.09-GHz repetition rate, delivering up to 2.5 W of average output power. This laser provides access to a new parameter space that could previously not be reached by solid-state lasers and, to the best of our knowledge, is the first modelocked solid-state Yb:YAG laser in the gigahertz regime. This is achieved by introducing a single additional intracavity element, specifically a nonlinear birefringent YVO4 crystal, for soliton formation, polarization selection, and cavity intensity clamping. The isotropic pump absorption in Yb:YAG allows for stable and low-noise operation with multimode fiber pumping. This laser is ideally suited as a seed source for many commercial high-power Yb-doped amplification systems operating at a center wavelength around 1.03 µm. The laser exhibits a high power per comb line of 5.0 mW which also makes it interesting for applications in frequency comb spectroscopy, especially if it is used to pump an optical parametric oscillator. We measure a relative intensity noise (RIN) of 0.03%, integrated from 1 Hz to 10 MHz. Furthermore, we show that the laser timing jitter for noise frequencies >2 kHz is fully explained by a power-dependent shift in the center wavelength of 0.38 nm/W due to the quasi-three-level laser gain material. The narrow gain bandwidth of Yb:YAG reduces this contribution to noise in comparison to other SESAM modelocked Yb-doped lasers.

Single-cavity dual-modelocked 2.36-µm laser.

We present the first dual-modelocked femtosecond oscillator operating beyond 2 µm wavelength. This new class of laser is based on a Cr:ZnS gain medium, an InGaSb SESAM for modelocking, and a two-surface reflective device for spatial duplexing of the two modelocked pulse trains (combs). The laser operates at 2.36 µm, and for each comb, we have achieved a FWHM spectral bandwidth of 30 nm, an average power of over 200 mW, and a pulse duration close to 200 fs. The nominal repetition rate is 242 MHz with a sufficiently large repetition rate difference of 4.17 kHz. We also found that the laser is able to produce stable modelocked pulses over a wide range of output powers. This result represents a significant step towards realizing dual-comb applications directly above 2 µm using a single free-running laser.

Free-running Yb:KYW dual-comb oscillator in a MOPA architecture.

Single-cavity dual-combs comprise a rapidly emerging technology platform suitable for a wide range of applications like optical ranging, equivalent time sampling, and spectroscopy. However, it remains a challenging task to develop a dual-comb system that exhibits low relative frequency fluctuations to allow for comb line resolved measurements, while simultaneously offering high average power and short pulse durations. Here we combine a passively cooled and compact dual-comb solid-state oscillator with a pair of core-pumped Yb-fiber-based amplifiers in a master-oscillator power-amplifier (MOPA) architecture. The Yb:KYW oscillator operates at 250 MHz and uses polarization multiplexing for dual-comb generation. To the best of our knowledge, this is the first demonstration of a single-cavity dual-comb based on this gain material. As the pulse timing characteristics inherent to the oscillator are preserved in the amplification process, the proposed hybrid approach leverages the benefit of both the ultra-low noise solid-state laser and the advantages inherent to fiber amplifier systems such as straight-forward power scaling. The amplifier is optimized for minimal pulse broadening while still providing significant amplification and spectral broadening. We obtain around 1 W of power per output beam with pulses then compressed down to sub-90 fs using a simple grating compressor, while no pre-chirping or other dispersion management is needed. The full-width half-maximum (FWHM) of the radio-frequency comb teeth is 700 Hz for a measurement duration of 100 ms, which is much less than the typical repetition rate difference, making this passively stable source well-suited for indefinite coherent signal averaging via computational phase tracking.

Gene Expression and Pathway Analysis in Rat Brain and Liver After Exposure to Royal Demolition Explosive (Hexahydro-1,3,5-Trinitro-1,3,5-Triazine).

The nitramine explosive, hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is associated with acute and chronic toxicity in mammals and targets both the central nervous system and liver. After a single oral dose of RDX in male rats, the systemic distribution of RDX and the toxicodynamic response was measured using clinical chemistry and Affymetrix Rat Genome® 230 2.0 gene expression arrays, respectively. Nominal doses of 0, 9 and 36 mg/kg pure RDX were administered to animals followed by liver, cerebral cortex, and hippocampus gene expression analysis at 0, 3.5, 24, and 48 hours. RDX quickly entered the liver and brain, increasing up to 24 hours. For the 36 mg/kg dose, RDX was still measurable in liver and brain at 48 hours, but was non-detectible for the 9 mg/kg dose. At 3.5 hours, the time within which most convulsions reportedly occur after RDX ingestion, the hippocampus displayed the highest response for both gene expression and pathways, while the cortex was relatively non-responsive. The top 2 impacted pathways, primarily involved in neurotransmission, were the GABAergic and glutamatergic pathways. High numbers of genes also responded to RDX in the liver with P450 metabolism pathways significantly involved. Compared to the liver, the hippocampus displayed more consistent biological effects across dose and time with neurotransmission pathways predominating. Overall, based on gene expression data, RDX responses were high in both the hippocampus and liver, but were minimal in the cerebral cortex. These results identify the hippocampus as an important target for RDX based on gene expression.

Influence of disk aberrations on high-power thin-disk laser cavities.

We present a systematic study on the influence of thin-disk aberrations on the performance of thin-disk laser oscillators. To evaluate these effects, we have developed a spatially resolved numerical model supporting arbitrary phase profiles on the intracavity components that estimates the intracavity beam shape and the output power of thin-disk laser oscillators. By combining this model with the experimentally determined phase profile of the thin-disk (measured with interferometry), we can predict the operation mode of high-power thin-disk lasers, including mode degradation, higher-order mode coupling, and stability zone shrinking, all of which are in good agreement with experiment. Our results show that one of the main mechanisms limiting the performance is the small deviation of the disk's phase profile from perfect radial symmetry. This result is an important step to scaling modelocked thin-disk oscillators to the kW-level and will be important in the design of future active multi-pass cavity arrangements.

Dynamic and precise long-distance ranging using a free-running dual-comb laser.

Long-distance ranging is a crucial tool for both industrial and scientific applications. Laser-based distance metrology offers unprecedented precision making it the ideal approach for many deployments. In particular, dual-comb ranging is favorable due to its inherently high precision and sampling rate. To make high-performance long-range dual-comb LiDAR more accessible by reducing both cost and complexity, here we demonstrate a fiber-based dual-comb LiDAR frontend combined with a free-running diode-pumped solid-state dual-comb laser that allows for sub-µm measurement precision while offering a theoretical ambiguity range of more than 200 km. Our system simultaneously measures distance with the role of each comb interchanged, thereby enabling Vernier-based determination of the number of ambiguity ranges. As a proof-of-principle experiment, we measure the distance to a moving target over more than 10 m with sub-µm precision and high update rate, corresponding to a relative precision of 10-7. For a static target at a similar distance, we achieve an instantaneous precision of 0.29 µm with an update time of 1.50 ms. With a longer averaging time of 200 ms, we reach a precision of around 33 nm, which corresponds to a relative precision of about 3·10-9 with a time-of-flight-based approach.

High-power low-noise 2-GHz femtosecond laser oscillator at 2.4†µm.

Femtosecond lasers with high repetition rates are attractive for spectroscopic applications with high sampling rates, high power per comb line, and resolvable lines. However, at long wavelengths beyond 2 µm, current laser sources are either limited to low output power or repetition rates below 1 GHz. Here we present an ultrafast laser oscillator operating with high output power at multi-GHz repetition rate. The laser produces transform-limited 155-fs pulses at a repetition rate of 2 GHz, and an average power of 0.8 W, reaching up to 0.7 mW per comb line at the center wavelength of 2.38 µm. We have achieved this milestone via a Cr2+-doped ZnS solid-state laser modelocked with an InGaSb/GaSb SESAM. The laser is stable over several hours of operation. The integrated relative intensity noise is 0.15% rms for [10 Hz, 100 MHz], and the laser becomes shot noise limited (-160 dBc/Hz) at frequencies above 10 MHz. Our timing jitter measurements reveal contributions from pump laser noise and relaxation oscillations, with a timing jitter of 100 fs integrated over [3 kHz, 100 MHz]. These results open up a path towards fast and sensitive spectroscopy directly above 2 µm.

Absolute SESAM characterization via polarization-resolved non-collinear equivalent time sampling.

Semiconductor saturable absorber mirrors (SESAMs) have enabled a wide variety of modelocked laser systems, which makes measuring their nonlinear properties an important step in laser design. Here, we demonstrate complete characterization of SESAMs using an equivalent time sampling apparatus. The light source is a free-running dual-comb laser, which produces a pair of sub-150-fs modelocked laser outputs at 1051 nm from a single cavity. The average pulse repetition rate is 80.1 MHz, and the full time window is scanned at 240 Hz. Cross-correlation between the beams is used to calibrate the time axis of the measurements, and we use a non-collinear pump-probe geometry on the sample. The measurements enable fast and robust determination of all the nonlinear reflectivity and recovery time parameters of the devices from a single setup, and show good agreement with conventional nonlinear reflectivity measurements. We compare measurements to a rate equation model, showing good agreement up to high pulse fluence values and revealing that the samples tested exhibit a slightly slower recovery at higher fluence values. Lastly, we examine the polarization dependence of the reflectivity, revealing a reduced rollover if cross-polarized beams are used or if the sample is oriented optimally around the beam axis.

Isotropization of a Rotating and Longitudinally Expanding ϕ4 Scalar System.

We study numerically the evolution of an expanding system of scalar fields. The initial configuration is non-isotropic and rotating. We calculate the energy-momentum tensor and angular momentum vector of the system. We compare the time scales associated with the isotropization of the transverse and longitudinal pressures, and the decay of the initial angular momentum. We show that even a fairly large initial angular momentum decays significantly faster than the pressure anisotropy.

Dual-comb ranging with frequency combs from single cavity free-running laser oscillators.

Laser ranging (LIDAR) with dual optical frequency combs enables high-resolution distance measurements over long ranges with fast update rates. However, the high complexity of stabilized dual optical frequency comb systems makes it challenging to use this technique in industrial applications. To address this issue, here we demonstrate laser ranging directly from the output of both a free-running dual-comb diode-pumped semiconductor and solid-state laser oscillator. Dual-comb operation from a single cavity is achieved via polarization duplexing with intracavity birefringent crystals. We perform ranging experiments with two implementations of this scheme: a modelocked integrated external cavity surface-emitting laser (MIXSEL) and a Yb:CaF2 solid-state laser. For these proof of principle demonstrations, we measure the distance to a moving mirror mounted on a home-made shaker. The MIXSEL laser has a repetition rate of 2.736 GHz and a repetition rate difference of 52 kHz, and yields a measurement resolution of 1.36 µm. The Yb:CaF2 laser has a repetition rate of 137 MHz and a repetition rate difference of 952 Hz, and yields a measurement resolution of 0.55 µm. In both cases the resolution is inferred by a parallel measurement with a HeNe interferometer. These results represent the first laser ranging with free-running dual-comb solid-state oscillators. With further optimization, resolution well below 1 µm and range well above 1 km are expected with this technique.

Picosecond ultrasonics with a free-running dual-comb laser.

We present a free-running 80-MHz dual-comb polarization-multiplexed solid-state laser which delivers 1.8 W of average power with 110-fs pulse duration per comb. With a high-sensitivity pump-probe setup, we apply this free-running dual-comb laser to picosecond ultrasonic measurements. The ultrasonic signatures in a semiconductor multi-quantum-well structure originating from the quantum wells and superlattice regions are revealed and discussed. We further demonstrate ultrasonic measurements on a thin-film metalized sample and compare these measurements to ones obtained with a pair of locked femtosecond lasers. Our data show that a free-running dual-comb laser is well-suited for picosecond ultrasonic measurements and thus it offers a significant reduction in complexity and cost for this widely adopted non-destructive testing technique.

Impact of excessive alcohol abuse on age prediction using the VISAGE enhanced tool for epigenetic age estimation in blood.

DNA methylation-based clocks provide the most accurate age estimates with practical implications for clinical and forensic genetics. However, the effects of external factors that may influence the estimates are poorly studied. Here, we evaluated the effect of alcohol consumption on epigenetic age prediction in a cohort of extreme alcohol abusers. Blood samples from deceased alcohol abusers and age- and sex-matched controls were analyzed using the VISAGE enhanced tool for age prediction from somatic tissues that enables examination of 44 CpGs within eight age markers. Significantly altered DNA methylation was recorded for alcohol abusers in MIR29B2CHG. This resulted in a mean predicted age of 1.4 years higher compared to the controls and this trend increased in older individuals. The association of alcohol abuse with epigenetic age acceleration, as determined by the prediction analysis performed based on MIR29B2CHG, was small but significant (ß = 0.190; P-value = 0.007). However, the observed alteration in DNA methylation of MIR29B2CHG had a non-significant effect on age estimation with the VISAGE age prediction model. The mean absolute error in the alcohol-abusing cohort was 3.1 years, compared to 3.3 years in the control group. At the same time, upregulation of MIR29B2CHG expression may have a biological function, which merits further studies.

Changes in Pain Medicine Training Programs Associated With COVID-19: Survey Results.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic is a public health crisis of unprecedented proportions that has altered the practice of medicine. The pandemic has required pain clinics to transition from in-person visits to telemedicine, postpone procedures, and cancel face-to-face educational sessions. There are no data on how fellowship programs have adapted. METHODS: A 17-question survey was developed covering topics including changes in education, clinical care, and psychological stress due to the COVID pandemic. The initial survey was hosted by Qualtrics Inc and disseminated by the Association of Pain Program Directors on April 10, 2020, to program directors at Accreditation Council for Graduate Medical Education (ACGME)-accredited fellowships. Results are reported descriptively and stratified by COVID infection rate, which was calculated from Centers for Disease Control and Prevention data on state infections, and census data. RESULTS: Among 107 surveys distributed, 70 (65%) programs responded. Twenty-nine programs were located in states in the upper tertile for per capita infection rates, 17 in the middle third, and 23 in the lowest tertile. Nearly all programs (93%) reported a decreased workload, with 11 (16%) reporting a dramatic decrease (only urgent or emergent cases). Just more than half of programs had either already deployed (14%) or credentialed (39%) fellows to provide nonpain care. Higher state infection rates were significantly associated with reduced clinical demand (Rs = 0.31, 95% confidence interval [CI], 0.08-0.51; P = .011) and redeployment of fellows to nonpain areas (Rs = 0.30, 95% CI, 0.07-0.50; P = .013). Larger program size but not infection rate was associated with increased perceived anxiety level of trainees. CONCLUSIONS: We found a shift to online alternatives for clinical care and education, with correlations between per capita infection rates, and clinical care demands and redeployment, but not with overall trainee anxiety levels. It is likely that medicine in general, and pain medicine in particular, will change after COVID-19, with greater emphasis on telemedicine, virtual education, and greater national and international cooperation. Physicians should be prepared for these changes.

BLOOD LACTATE CONCENTRATIONS IN EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA) FOLLOWING CAPTURE BY A CANINE SEARCH TEAM.

Studies to assess wildlife health commonly evaluate clinical pathology changes, immune responses, pathogen presence, and contaminant exposure, but novel modalities are needed to characterize the unique physiologic responses of reptiles. Lactate is an indicator of hypoperfusion and/or anaerobic respiration and can be quickly and easily measured using a point-of-care analyzer. This study evaluated baseline blood lactate concentrations in free-living eastern box turtles (Terrapene carolina carolina, n = 116) using a point of care analyzer and then determined the effect of handling time, physical examination (PE) abnormalities, and quantitative polymerase chain reaction pathogen detection (Terrapene herpesvirus 1, Mycoplasma sp., Terrapene adenovirus) on lactate concentrations. Blood lactate concentrations were higher in turtles with Terrapene herpesvirus 1 (n = 11), quiet mentation, and increased packed cell volume (P < 0.05). Lactate concentrations increased between initial capture and PE, with peak values reaching 129 min after capture. Lactate at PE was positively associated with baseline lactate concentrations. Turtles with Terrapene herpesvirus 1 may have alterations in blood flow, oxygen delivery, or activity patterns, driving increases in baseline lactate. Increased handling time likely leads to more escape behaviors and/or breath holding, causing turtles to undergo anaerobic metabolism and raising lactate concentrations. Overall, lactate measured by a point of care analyzer shows variability caused by capture and health factors in eastern box turtles and may be a useful adjunctive diagnostic test in this species after full methodologic validation.

Contrasting signatures of introgression in North American box turtle (Terrapene spp.) contact zones.

Hybridization occurs differentially across the genome in a balancing act between selection and migration. With the unprecedented resolution of contemporary sequencing technologies, selection and migration can now be effectively quantified such that researchers can identify genetic elements involved in introgression. Furthermore, genomic patterns can now be associated with ecologically relevant phenotypes, given availability of annotated reference genomes. We do so in North American box turtles (Terrapene) by deciphering how selection affects hybrid zones at the interface of species boundaries and identifying genetic regions potentially under selection that may relate to thermal adaptations. Such genes may impact physiological pathways involved in temperature-dependent sex determination, immune system functioning and hypoxia tolerance. We contrasted these patterns across inter- and intraspecific hybrid zones that differ temporally and biogeographically. We demonstrate hybridization is broadly apparent in Terrapene, but with observed genomic cline patterns corresponding to species boundaries at loci potentially associated with thermal adaptation. These loci display signatures of directional introgression within intraspecific boundaries, despite a genome-wide selective trend against intergrades. In contrast, outlier loci for interspecific comparisons exhibited evidence of being under selection against hybrids. Importantly, adaptations coinciding with species boundaries in Terrapene overlap with climatic boundaries and highlight the vulnerability of these terrestrial ectotherms to anthropogenic pressures.

Femtosecond dual-comb Yb:CaF2 laser from a single free-running polarization-multiplexed cavity for optical sampling applications.

Dual optical frequency combs are an appealing solution to many optical measurement techniques due to their high spectral and temporal resolution, high scanning speed, and lack of moving parts. However, industrial and field-deployable applications of such systems are limited due to a high-cost factor and intricacy in the experimental setups, which typically require a pair of locked femtosecond lasers. Here, we demonstrate a single oscillator which produces two mode-locked output beams with a stable repetition rate difference. We achieve this via inserting two 45°-cut birefringent crystals into the laser cavity, which introduces a repetition rate difference between the two polarization states of the cavity. To mode-lock both combs simultaneously, we use a semiconductor saturable absorber mirror (SESAM). We achieve two simultaneously operating combs at 1050 nm with 175-fs duration, 3.2-nJ pulses and an average power of 440 mW in each beam. The average repetition rate is 137 MHz, and we set the repetition rate difference to 1 kHz. This laser system, which is the first SESAM mode-locked femtosecond solid-state dual-comb source based on birefringent multiplexing, paves the way for portable and high-power femtosecond dual-combs with flexible repetition rate. To demonstrate the utility of the laser for applications, we perform asynchronous optical sampling (ASOPS) on semiconductor thin-film structures with the free-running laser system, revealing temporal dynamics from femtosecond to nanosecond time scales.

Performance scaling of a 10-GHz solid-state laser enabling self-referenced CEO frequency detection without amplification.

A simple and compact straight-cavity laser oscillator incorporating a cascaded quadratic nonlinear crystal and a semiconductor saturable absorber mirror (SESAM) can deliver stable femtosecond modelocking at high pulse repetition rates >10 GHz. In this paper, we experimentally investigate the influence of intracavity dispersion, pump brightness, and cavity design on modelocking with high repetition rates, and use the resulting insights to demonstrate a 10.4-GHz straight-cavity SESAM-modelocked Yb:CALGO laser delivering 108-fs pulses with 812 mW of average output power. This result represents a record-level performance for diode-pumped femtosecond oscillators with repetition rates above 10 GHz. Using the oscillator output without any optical amplification, we demonstrate coherent octave-spanning supercontinuum generation (SCG) in a silicon nitride waveguide. Subsequent f-to-2f interferometry with a periodically poled lithium niobate waveguide enables the detection of a strong carrier-envelope offset (CEO) beat note with a 33-dB signal-to-noise ratio.