Polysaccharide monooxygenases (PMOs) are secreted metalloenzymes that catalyze the oxidative degradation of polysaccharides in a copper-, oxygen-, and reductant-dependent manner. Cellulose-active fungal PMOs degrade cellulosic substrates to be utilized as a carbon source for fungal growth. To gain insight into the PMO mechanism, the role of conserved residues in the copper coordination sphere was investigated. Here, we report active-site hydrogen-bonding motifs in the secondary copper coordination sphere of MtPMO3*, a C1-oxidizing PMO from the ascomycete fungus Myceliophthora thermophila. A series of point substitutions that disrupt this conserved network are used to interrogate its function. Activity assays, in conjunction with EPR spectroscopy, demonstrate that residues H161 and Q167 are involved in stabilizing bound oxygen, and H161 appears to play a role in proton transfer. Additionally, Q167 increases the ligand donor strength of Y169 to the copper via a hydrogen-bonding interaction. Altogether, H161 and Q167 are important for oxygen activation, and the results are suggestive of a copper-oxyl active intermediate.

Purpose

To assess the utility of furosemide diuresis and the role of an improved scatter correction algorithm in reducing scatter artifact severity on Ga-68- Prostate-specific membrane antigen (PSMA)-11 positron emission tomography (PET).

Materials and methods

A total of 139 patients underwent Ga-68-PSMA-11 PET imaging for prostate cancer: 47 non-time-of-flight (non-TOF) PET/computed tomography, 51 PET/magnetic resonance imaging (MRI) using the standard TOF scatter correction algorithm, and 41 PET/MRI using an improved TOF scatter correction algorithm. Whole-body PET acquisitions were subdivided into 3 regions: around kidneys; between kidneys and bladder; and around bladder. The images were reviewed, and scatter artifact severity was rated using a Likert-type scale.

Results

The worst scatter occurred when using non-TOF scatter correction without furosemide, where 42.1% of patients demonstrated severe scatter artifacts in 1 or more regions. Improved TOF scatter correction resulted in the smallest percentage of studies with severe scatter (6.5%). Scatter ratings by region were lowest using improved TOF scatter correction. Furosemide reduced mean scatter severity when using non-TOF and standard TOF.

Conclusions

Both furosemide and scatter correction algorithm play a role in reducing scatter in PSMA PET. Improved TOF scatter correction resulted in the lowest scatter severity.

¹⁵O-H₂O PET imaging is an accurate method to measure cerebral blood flow (CBF) but it requires an arterial input function (AIF). Historically, image-derived AIF estimation suffers from low temporal resolution, spill-in, and spill-over problems. Here, we optimized tracer dose on a time-of-flight PET/MR according to the acquisition-specific noise-equivalent count rate curve. An optimized dose of 850 MBq of ¹⁵O-H₂O was determined, which allowed sufficient counts to reconstruct a short time-frame PET angiogram (PETA) during the arterial phase. This PETA enabled the measurement of the extent of spill-over, while an MR angiogram was used to measure the true arterial volume for AIF estimation. A segment of the high cervical arteries outside the brain was chosen, where the measured spill-in effects were minimal. CBF studies were performed twice with separate [15O]-H₂O injections in 10 healthy subjects, yielding values of 88 ± 16, 44 ± 9, and 58 ± 11 mL/min/100 g for gray matter, white matter, and whole brain, with intra-subject CBF differences of 5.0 ± 4.0%, 4.1 ± 3.3%, and 4.5 ± 3.7%, respectively. A third CBF measurement after the administration of 1 g of acetazolamide showed 35 ± 23%, 29 ± 20%, and 33 ± 22% increase in gray matter, white matter, and whole brain, respectively. Based on these findings, the proposed noninvasive AIF method provides robust CBF measurement with ¹⁵O-H₂O PET.

Catalytic modules of assembly-line polyketide synthases (PKSs) have previously been observed in two very different conformations-an "extended" architecture and an "arch-shaped" architecture-although the catalytic relevance of neither has been directly established. By the use of a fully human naïve antigen-binding fragment (F_ab) library, a high-affinity antibody was identified that bound to the extended conformation of a PKS module, as verified by X-ray crystallography and tandem size-exclusion chromatography-small-angle X-ray scattering (SEC-SAXS). Kinetic analysis proved that this antibody-stabilized module conformation was fully competent for catalysis of intermodular polyketide chain translocation as well as intramodular polyketide chain elongation and functional group modification of a growing polyketide chain. Thus, the extended conformation of a PKS module is fully competent for all of its essential catalytic functions.

RadioAstron space-ground very long baseline interferometry observations of the pulsar B0950+08, conducted with the 10 m Space Radio Telescope in conjunction with the Arecibo 300 m telescope and the Westerbork Synthesis Radio Telescope at a frequency of 324 MHz were analyzed in order to investigate plasma inhomogeneities in the direction of this nearby pulsar. The observations were conducted at a spacecraft distance of 330, 000 km, resulting in a projected baseline of 220, 000 km, providing the greatest angular resolution ever achieved at meter wavelengths. Our analysis is based on fundamental behavior of structure and coherence functions. We find that the pulsar shows scintillation on two frequency scales, both much less than the observing frequency, but modulation is less than 100%. We infer that the scattering is weak, but a refracting wedge disperses the scintillation pattern. The refraction angle of this "cosmic prism" is measured as θ0 = 1.1-4.4 mas, with the refraction direction being approximately perpendicular to the observer velocity. We show that the observed parameters of scintillation effects indicate that two plasma layers lie along the line of sight to the pulsar, at distances of 4.4-16.4 pc and 26-170 pc, and traveling in different directions relative to the line of sight. Spectra of turbulence for the two layers are found to follow a power law with the indices γ1 = γ2 = 3.00 ± 0.08, significantly different from the index expected for a Kolmogorov spectrum of turbulence, γ = 11/3.

FRB 190608 was detected by ASKAP and localized to a spiral galaxy at $z_{host}=0.11778$ in the SDSS footprint. The burst has a large dispersion measure ($DM_{FRB}=339.8$ $pc/cm^3$) compared to the expected cosmic average at its redshift. It also has a large rotation measure ($RM_{FRB}=353$ $rad/m^2$) and scattering timescale ($\tau=3.3$ $ms$ at $1.28$ $GHz$). Chittidi et al (2020) perform a detailed analysis of the ultraviolet and optical emission of the host galaxy and estimate the host DM contribution to be $110\pm 37$ $pc/cm^3$. This work complements theirs and reports the analysis of the optical data of galaxies in the foreground of FRB 190608 to explore their contributions to the FRB signal. Together, the two manuscripts delineate an observationally driven, end-to-end study of matter distribution along an FRB sightline; the first study of its kind. Combining KCWI observations and public SDSS data, we estimate the expected cosmic dispersion measure $DM_{cosmic}$ along the sightline to FRB 190608. We first estimate the contribution of hot, ionized gas in intervening virialized halos ($DM_{halos} \approx 7-28$ $pc/cm^3$). Then, using the Monte Carlo Physarum Machine (MCPM) methodology, we produce a 3D map of ionized gas in cosmic web filaments and compute the DM contribution from matter outside halos ($DM_{IGM} \approx 91-126$ $pc/cm^3$). This implies a greater fraction of ionized gas along this sightline is extant outside virialized halos. We also investigate whether the intervening halos can account for the large FRB rotation measure and pulse width and conclude that it is implausible. Both the pulse broadening and the large Faraday rotation likely arise from the progenitor environment or the host galaxy.

Purpose

There are several important positron emission tomography (PET) imaging scenarios that require imaging with very low photon statistics, for which both quantitative accuracy and visual quality should not be neglected. For example, PET imaging with the low photon statistics is closely related to active efforts to significantly reduce radiation exposure from radiopharmaceuticals. We investigated two examples of low-count PET imaging: (a) imaging [⁹⁰Y]microsphere radioembolization that suffers the very small positron emission fraction of Y-90's decay processes, and (b) cancer imaging with [⁶⁸Ga]citrate with uptake time of 3-4 half-lives, necessary for visualizing tumors. In particular, we investigated a type of penalized likelihood reconstruction algorithm, block sequential regularized expectation maximization (BSREM), for improving both image quality and quantitative accuracy of these low-count PET imaging cases.

Procedures

The NEMA/IEC Body phantom filled with aqueous solution of Y-90 or Ga-68 was scanned to mimic the low-count scenarios of corresponding patient data acquisitions on a time-of-flight (TOF) PET/magnetic resonance imaging system. Contrast recovery, background variation, and signal-to-noise ratio were evaluated in different sets of count densities using both conventional TOF ordered subset expectation (TOF-OSEM) and TOF-BSREM algorithms. The regularization parameter, beta, in BSREM that controls the tradeoff between image noise and resolution was evaluated to find a value for improved confidence in image interpretation. Visual quality assessment of the images obtained from patients administered with [⁶⁸Ga]citrate (n = 6) was performed. We also made preliminary visual image quality assessment for one patient with [⁹⁰Y]microspheres. In Y-90 imaging, the effect of 511-keV energy window selection for minimizing the number of random events was also evaluated.

Results

Quantitatively, phantom images reconstructed with TOF-BSREM showed improved contrast recovery, background variation, and signal-to-noise ratio values over images reconstructed with TOF-OSEM. Both phantom and patient studies of delayed imaging of [⁶⁸Ga]citrate show that TOF-BSREM with beta = 500 gives the best tradeoff between image noise and image resolution based on visual assessment by the readers. The NEMA-IQ phantom study with [⁹⁰Y]microspheres shows that the narrow energy window (460-562 keV) recovers activity concentrations in small spheres better than the regular energy window (425-650 keV) with the beta value of 2000 using the TOF-BSREM algorithm. For the images obtained from patients with [⁶⁸Ga]citrate using TOF-BSREM with beta = 500, the visual analogue scale (VAS) was improved by 17 % and the Likert score was increased by 1 point on average, both in comparison to corresponding scores for images reconstructed using TOF-OSEM.

Conclusion

Our investigation shows that the TOF-BSREM algorithm improves the image quality and quantitative accuracy in low-count PET imaging scenarios. However, the beta value in this algorithm needed to be adjusted for each radiopharmaceutical and counting statistics at the time of scans.

Continued timing observations of the double pulsar PSR J0737-3039A/B, which consists of two active radio pulsars (A and B) that orbit each other with a period of 2.45 h in a mildly eccentric (e=0.088) binary system, have led to large improvements in the measurement of relativistic effects in this system. With a 16-yr data span, the results enable precision tests of theories of gravity for strongly self-gravitating bodies and also reveal new relativistic effects that have been expected but are now observed for the first time. These include effects of light propagation in strong gravitational fields which are currently not testable by any other method. In particular, we observe the effects of retardation and aberrational light bending that allow determination of the spin direction of the pulsar. In total, we detect seven post-Keplerian parameters in this system, more than for any other known binary pulsar. For some of these effects, the measurement precision is now so high that for the first time we have to take higher-order contributions into account. These include the contribution of the A pulsar's effective mass loss (due to spin-down) to the observed orbital period decay, a relativistic deformation of the orbit, and the effects of the equation of state of superdense matter on the observed post-Keplerian parameters via relativistic spin-orbit coupling. We discuss the implications of our findings, including those for the moment of inertia of neutron stars, and present the currently most precise test of general relativity's quadrupolar description of gravitational waves, validating the prediction of general relativity at a level of 1.3×10-4 with 95% confidence. We demonstrate the utility of the double pulsar for tests of alternative theories of gravity by focusing on two specific examples and also discuss some implications of the observations for studies of the interstellar medium and models for the formation of the double pulsar system. Finally, we provide context to other types of related experiments and prospects for the future.

NANOG (from Irish mythology Tír na nÓg) transcription factor plays a central role in maintaining pluripotency, cooperating with OCT4 (also known as POU5F1 or OCT3/4), SOX2, and other pluripotency factors. Although the physiological roles of the NANOG protein have been extensively explored, biochemical and biophysical properties in relation to its structural analysis are poorly understood. Here we determined the crystal structure of the human NANOG homeodomain (hNANOG HD) bound to an OCT4 promoter DNA, which revealed amino acid residues involved in DNA recognition that are likely to be functionally important. We generated a series of hNANOG HD alanine substitution mutants based on the protein-DNA interaction and evolutionary conservation and determined their biological activities. Some mutant proteins were less stable, resulting in loss or decreased affinity for DNA binding. Overexpression of the orthologous mouse NANOG (mNANOG) mutants failed to maintain self-renewal of mouse embryonic stem cells without leukemia inhibitory factor. These results suggest that these residues are critical for NANOG transcriptional activity. Interestingly, one mutant, hNANOG L122A, conversely enhanced protein stability and DNA-binding affinity. The mNANOG L122A, when overexpressed in mouse embryonic stem cells, maintained their expression of self-renewal markers even when retinoic acid was added to forcibly drive differentiation. When overexpressed in epiblast stem cells or human induced pluripotent stem cells, the L122A mutants enhanced reprogramming into ground-state pluripotency. These findings demonstrate that structural and biophysical information on key transcriptional factors provides insights into the manipulation of stem cell behaviors and a framework for rational protein engineering.

We demonstrate a novel detection method for the cyclotron resonance frequency of an electron plasma in a Penning-Malmberg trap. With this technique, the electron plasma is used as an in situ diagnostic tool for the measurement of the static magnetic field and the microwave electric field in the trap. The cyclotron motion of the electron plasma is excited by microwave radiation and the temperature change of the plasma is measured non-destructively by monitoring the plasma's quadrupole mode frequency. The spatially resolved microwave electric field strength can be inferred from the plasma temperature change and the magnetic field is found through the cyclotron resonance frequency. These measurements were used extensively in the recently reported demonstration of resonant quantum interactions with antihydrogen. © 2014 IOP Publishing and Deutsche Physikalische Gesellschaft.