A photometric survey for Lyalpha-HeII dual emitters: Searching for Population III stars in high-redshift galaxies

We present a new photometric search for high-z galaxies hosting Population III (PopIII) stars based on deep intermediate-band imaging observations obtained in the Subaru Deep Field (SDF), by using Suprime-Cam on the Subaru Telescope. By combining our new data with the existing broad-band and narrow-band data, we searched for galaxies which emit strongly both in Ly_alpha and in HeII 1640 (``dual emitters'') that are promising candidates for PopIII-hosting galaxies, at 3.93<z<4.01 and 4.57<z<4.65. Although we found 10 ``dual emitters'', most of them turn out to be [OII]-[OIII] dual emitters or H_beta-(H_alpha+[NII]) dual emitters at z<1, as inferred from their broad-band colors and from the ratio of the equivalent widths. No convincing candidate of Ly_alpha-HeII dual emitter of SFR_PopIII > 2 Msun/yr was found by our photometric search in 4.03 x 10^5 Mpc^3 in the SDF. This result disfavors low feedback models for PopIII star clusters, and implies an upper-limit of the PopIII SFR density of SFRD_PopIII < 5 x 10^-6 Msun/yr/Mpc^3. This new selection method to search for PopIII-hosting galaxies should be useful in future narrow-band surveys to achieve the first observational detection of PopIII-hosting galaxies at high redshifts.Comment: 24 pages, 10 figures, accepted for publication in Ap

A Census of Star-Forming Galaxies at z = 1-3 in the Subaru Deep Field

Several UV and near-infrared color selection methods have identified galaxies at z = 1-3. Since each method suffers from selection biases, we have applied three leading techniques (Lyman break, BX/BM, and BzK selection) simultaneously in the Subaru Deep Field. This field has reliable ({\Delta}z/(1 + z) = 0.02--0.09) photometric redshifts for ~53,000 galaxies from 20 bands (1500{\AA}--2.2{\mu}m). The BzK, LBG, and BX/BM samples suffer contamination from z<1 interlopers of 6%, 8%, and 20%, respectively. Around the redshifts where it is most sensitive (z~1.9 for star-forming BzK, z~1.8 for z~2 LBGs, z~1.6 for BM, and z~2.3 for BX), each technique finds 60-80% of the census of the three methods. In addition, each of the color techniques shares 75-96% of its galaxies with another method, which is consistent with previous studies that adopt identical criteria on magnitudes and colors. Combining the three samples gives a comprehensive census that includes ~90% of z-phot = 1-3 galaxies, using standard magnitude limits similar to previous studies. In fact, we find that among z = 1-2.5 galaxies in the color selection census, 81-90% of them can be selected by just combining the BzK selection with one of the UV techniques (z~2 LBG or BX and BM). The average galaxy stellar mass, reddening and SFRs all decrease systematically from the sBzK population to the LBGs, and to the BX/BMs. The combined color selections yield a total cosmic SFR density of 0.18 $\pm$ 0.03 M_sun yr^{-1} Mpc^{-3} for K_AB <= 24. We find that 65% of the star formation is in galaxies with E(B-V) > 0.25 mag, even though they are only one-fourth of the census by number.Comment: 26 pages, 30 figures, 9 tables, emulateapj format. Modified to match the final ApJ versio

Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN− Ligands Derive from Tyrosine

[FeFe] hydrogenases are promising catalysts for producing hydrogen as a sustainable fuel and chemical feedstock, and they also serve as paradigms for biomimetic hydrogen-evolving compounds. Hydrogen formation is catalyzed by the H-cluster, a unique iron-based cofactor requiring three carbon monoxide (CO) and two cyanide (CN−) ligands as well as a dithiolate bridge. Three accessory proteins (HydE, HydF, and HydG) are presumably responsible for assembling and installing the H-cluster, yet their precise roles and the biosynthetic pathway have yet to be fully defined. In this report, we describe effective cell-free methods for investigating H-cluster synthesis and [FeFe] hydrogenase activation. Combining isotopic labeling with FTIR spectroscopy, we conclusively show that each of the CO and CN− ligands derive respectively from the carboxylate and amino substituents of tyrosine. Such in vitro systems with reconstituted pathways comprise a versatile approach for studying biosynthetic mechanisms, and this work marks a significant step towards an understanding of both the protein-protein interactions and complex reactions required for H-cluster assembly and hydrogenase maturation

High-Yield Expression of Heterologous [FeFe] Hydrogenases in Escherichia coli

BACKGROUND: The realization of hydrogenase-based technologies for renewable H(2) production is presently limited by the need for scalable and high-yielding methods to supply active hydrogenases and their required maturases. PRINCIPAL FINDINGS: In this report, we describe an improved Escherichia coli-based expression system capable of producing 8-30 mg of purified, active [FeFe] hydrogenase per liter of culture, volumetric yields at least 10-fold greater than previously reported. Specifically, we overcame two problems associated with other in vivo production methods: low protein yields and ineffective hydrogenase maturation. The addition of glucose to the growth medium enhances anaerobic metabolism and growth during hydrogenase expression, which substantially increases total yields. Also, we combine iron and cysteine supplementation with the use of an E. coli strain upregulated for iron-sulfur cluster protein accumulation. These measures dramatically improve in vivo hydrogenase activation. Two hydrogenases, HydA1 from Chlamydomonas reinhardtii and HydA (CpI) from Clostridium pasteurianum, were produced with this improved system and subsequently purified. Biophysical characterization and FTIR spectroscopic analysis of these enzymes indicate that they harbor the H-cluster and catalyze H(2) evolution with rates comparable to those of enzymes isolated from their respective native organisms. SIGNIFICANCE: The production system we describe will facilitate basic hydrogenase investigations as well as the development of new technologies that utilize these prolific H(2)-producing enzymes. These methods can also be extended for producing and studying a variety of oxygen-sensitive iron-sulfur proteins as well as other proteins requiring anoxic environments

Early mobilisation in critically ill COVID-19 patients: a subanalysis of the ESICM-initiated UNITE-COVID observational study

Background Early mobilisation (EM) is an intervention that may improve the outcome of critically ill patients. There is limited data on EM in COVID-19 patients and its use during the first pandemic wave. Methods This is a pre-planned subanalysis of the ESICM UNITE-COVID, an international multicenter observational study involving critically ill COVID-19 patients in the ICU between February 15th and May 15th, 2020. We analysed variables associated with the initiation of EM (within 72 h of ICU admission) and explored the impact of EM on mortality, ICU and hospital length of stay, as well as discharge location. Statistical analyses were done using (generalised) linear mixed-effect models and ANOVAs. Results Mobilisation data from 4190 patients from 280 ICUs in 45 countries were analysed. 1114 (26.6%) of these patients received mobilisation within 72 h after ICU admission; 3076 (73.4%) did not. In our analysis of factors associated with EM, mechanical ventilation at admission (OR 0.29; 95% CI 0.25, 0.35; p = 0.001), higher age (OR 0.99; 95% CI 0.98, 1.00; p ≤ 0.001), pre-existing asthma (OR 0.84; 95% CI 0.73, 0.98; p = 0.028), and pre-existing kidney disease (OR 0.84; 95% CI 0.71, 0.99; p = 0.036) were negatively associated with the initiation of EM. EM was associated with a higher chance of being discharged home (OR 1.31; 95% CI 1.08, 1.58; p = 0.007) but was not associated with length of stay in ICU (adj. difference 0.91 days; 95% CI − 0.47, 1.37, p = 0.34) and hospital (adj. difference 1.4 days; 95% CI − 0.62, 2.35, p = 0.24) or mortality (OR 0.88; 95% CI 0.7, 1.09, p = 0.24) when adjusted for covariates. Conclusions Our findings demonstrate that a quarter of COVID-19 patients received EM. There was no association found between EM in COVID-19 patients' ICU and hospital length of stay or mortality. However, EM in COVID-19 patients was associated with increased odds of being discharged home rather than to a care facility. Trial registration ClinicalTrials.gov: NCT04836065 (retrospectively registered April 8th 2021)

Comparing this work to previous reports for the production of the <i>C. reinhardtii</i> HydA1 hydrogenase.

1<p>Specific activities for H₂ production rates are expressed in units of µmol H₂·min⁻¹·mg⁻¹ of HydA1.</p>2<p>Purification yields are mg of HydA1 isolated per liter of cell culture.</p>3<p>Iron content is expressed in units of Fe atoms per HydA1 peptide; NR, not reported.</p

Effects of iron and cysteine supplementation as well as the <i>iscR</i> deletion on active hydrogenase expression.

<p>Iron (2 mM ferric ammonium citrate) and cysteine (2 mM) were added to cultures as indicated. Methyl viologen reducing activities (µmol MV reduced·min⁻¹·mg⁻¹ total protein) of active [FeFe] hydrogenase in cell lysates from <i>E. coli</i> strains BL21(DE3) (gray bars) and BL21(DE3) <i>ΔiscR</i> (black bars). Hydrogenase activities were measured after 16–18 hrs of anaerobic HydA1 expression (A) and CpI expression (B). Hydrogenase activities for the <i>ΔiscR</i>-derived samples are indicated above the respective columns, and all activities are the average for n = 2 cultures ± standard deviations.</p

Biophysical characterization of purified [FeFe] hydrogenases produced in <i>Escherichia coli</i>.

<p>Data are the average from n = 3–6 cultures examined ± standard deviations.</p>†<p>Specific activities of purified hydrogenases were determined using methyl viologen (MV) or PetF ferredoxin as the electron donating or accepting substrates.</p