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Azobenzene derivatives, which show light‐induced reversible trans ↔ cis isomerization, have gained increasing attention in the area of protein science. p ‐(Phenylazo)‐L‐phenylalanine (Pap) was recently employed to enable the light‐controlled affinity purification of biosynthetic proteins as part of the Azo‐tag. Specific supramolecular complex formation with immobilized α ‐cyclodextrin ( α ‐CD) groups is mediated by the Pap side chain in its low‐energy trans ‐configuration, whereas photoisomerization to the cis ‐state leads to immediate dissociation. Here, we describe the X‐ray crystallographic analysis of super‐folder green fluorescent protein (sfGFP) displaying Pap at amino acid position 39 on its surface in complex with α ‐CD. While this experimental structure generally confirms the mode of host–guest interaction predicted by molecular modeling, there are two unexpected observations: (i) the conically shaped α ‐CD binds with its narrow end toward the aminoacyl moiety of Pap, despite appearing sterically more demanding, and (ii) the azobenzene side chain shows a considerably twisted conformation of its two phenyl rings, which contrasts with the fully coplanar arrangement usually anticipated for unmodified azobenzene and its chemical derivatives. Thus, this crystal structure of the photoswitchable noncanonical amino acid Pap (also known as AzoF or AzoPhe) provides valuable insight for future molecular engineering endeavors to endow proteins with light‐controllable functions.

The lack of a non-invasive test for malignant thyroid nodules makes the diagnosis of thyroid cancer (TC) challenging. Human galectin-3 (hGal3) has emerged as a promising target for medical TC imaging and diagnosis because of its exclusive overexpression in malignant thyroid tissues. We previously developed a human-chimeric αhGal3 Fab fragment derived from the rat monoclonal antibody (mAb) M3/38 with optimized clearance characteristics using PASylation technology. Here, we describe the elucidation of the hGal3 epitope recognized by mAb M3/38, X-ray crystallographic analysis of its complex with the chimeric Fab and, based on the three-dimensional structure, the rational humanization of the Fab by CDR grafting. Four CDR-grafted versions were designed using structurally most closely related fully human immunoglobulin V H /V L regions of which one—employing the acceptor framework regions of the HIV-1 neutralizing human antibody m66—showed the highest antigen affinity. By introducing two additional back-mutations to the rodent donor sequence, an affinity toward hGal3 indistinguishable from the chimeric Fab was achieved (K D  = 0.34 ± 0.02 nM in SPR). The PASylated humanized Fab was site-specifically labelled with the fluorescent dye Cy7 and applied for the immuno-histochemical staining of human tissue sections representative for different TCs. The same protein was conjugated with the metal chelator Dfo, followed by radiolabelling with 89 Zr(IV). The resulting protein tracer allowed the highly sensitive and specific PET/CT imaging of orthotopic tumors in mice, which was confirmed by quantitative analysis of radiotracer accumulation. Thus, the PASylated humanized αhGal3 Fab offers clinical potential for the diagnostic imaging of TC.

Most known archaeal DNA polymerases belong to the type B family, which also includes the DNA replication polymerases of eukaryotes, but maintain high fidelity at extreme conditions. We describe here the 2.5 angstrom resolution crystal structure of a DNA polymerase from the Archaea Thermococcus gorgonarius and identify structural features of the fold and the active site that are likely responsible for its thermostable function. Comparison with the mesophilic B type DNA polymerase gp43 of the bacteriophage RB69 highlights thermophilic adaptations, which include the presence of two disulfide bonds and an enhanced electrostatic complementarity at the DNA-protein interface. In contrast to gp43, several loops in the exonuclease and thumb domains are more closely packed; this apparently blocks primer binding to the exonuclease active site. A physiological role of this \"closed\" conformation is unknown but may represent a polymerase mode, in contrast to an editing mode with an open exonuclease site. This archaeal B DNA polymerase structure provides a starting point for structure-based design of polymerases or ligands with applications in biotechnology and the development of antiviral or anticancer agents.

The Front Cover illustrates the specific host/guest complex formation between the light‐switchable noncanonical amino acid /p/‐(phenylazo)‐L‐phenylalanine (Pap) in its low‐energy /trans/‐state and α‐cyclodextrin (α‐CD), which is elucidated by X‐ray crystallographic analysis of super‐folder green fluorescent protein (sfGFP) as carrier for the Pap side chain. Upon illumination with mild UV‐A light, Pap switches to the /cis/‐configuration and the complex dissociates, which can be utilized for the affinity purification of sfGFP(39Pap) on an α‐CD affinity column. More information can be found in the Research Article by Arne Skerra and co‐workers (DOI: 10.1002/open.202500471).

In this case series, investigators report a very rare but life-threatening sequela of ChAdOx1 nCoV-19 vaccination. Beginning 5 to 16 days after a first injection, some patients had symptoms consistent with thrombocytopenia, disseminated intravascular coagulation, and thromboses, including cerebral venous sinus thrombosis with catastrophic outcome. An anti–PF4 antibody capable of platelet activation appears to be the cause. Intravenous immune globulin may be therapeutic.

Mean age of air (AoA) is a diagnostic of transport along the stratospheric Brewer–Dobson circulation. While models consistently show negative trends, long-term time series (1975–2016) of AoA derived from observations show non-significant positive trends in mean AoA in the Northern Hemisphere. This discrepancy between observed and modelled mean AoA trends is still not resolved. There are uncertainties and assumptions required when deriving AoA from trace gas observations. At the same time, AoA from climate models is subject to uncertainties, too.In this paper, we focus on the uncertainties due to the parameter selection in the method that is used to derive mean AoA from SF6 measurements in . To correct for the non-linear increase in SF6 concentrations, a quadratic fit to the time series at the reference location, i.e. the tropical surface, is used. For this derivation, the width of the AoA distribution (age spectrum) has to be assumed. In addition, to choose the number of years the quadratic fit is performed for, the fraction of the age spectrum to be considered has to be assumed. Even though the uncertainty range due to all different aspects has already been taken into account for the total errors in the AoA values, the systematic influence of the parameter selection on AoA trends is described for the first time in the present study.For this, we use the EMAC (ECHAM MESSy Atmospheric Chemistry) climate model as a test bed, where AoA derived from a linear tracer is available as a reference and modelled age spectra exist to diagnose the actual spatial age spectra widths. The comparison of mean AoA from the linear tracer with mean AoA from a SF6 tracer shows systematic deviations specifically in the trends due to the selection of the parameters. However, for an appropriate parameter selection, good agreement for both mean AoA and its trend can be found, with deviations of about 1 % in mean AoA and 12 % in AoA trend.In addition, a method to derive mean AoA is evaluated that applies a convolution to the reference time series. The resulting mean AoA and its trend only depend on an assumption about the ratio of moments. Also in that case, it is found that the larger the ratio of moments, the more the AoA trend gravitates towards the negative. The linear tracer and SF6 AoA are found to agree within 0.3 % in the mean and 6 % in the trend.The different methods and parameter selections were then applied to the balloon-borne SF6 and CO2 observations. We found the same systematic changes in mean AoA trend dependent on the specific selection. When applying a parameter choice that is suggested by the model results, the AoA trend is reduced from 0.15 to 0.07 years per decade. It illustrates that correctly constraining those parameters is crucial for correct mean AoA and trend estimates and still remains a challenge in the real atmosphere.

The International Space Station (ISS) is a unique habitat for humans and microorganisms. Here, we report the results of the ISS experiment EXTREMOPHILES, including the analysis of microbial communities from several areas aboard at three time points. We assess microbial diversity, distribution, functional capacity and resistance profile using a combination of cultivation-independent analyses (amplicon and shot-gun sequencing) and cultivation-dependent analyses (physiological and genetic characterization of microbial isolates, antibiotic resistance tests, co-incubation experiments). We show that the ISS microbial communities are highly similar to those present in ground-based confined indoor environments and are subject to fluctuations, although a core microbiome persists over time and locations. The genomic and physiological features selected by ISS conditions do not appear to be directly relevant to human health, although adaptations towards biofilm formation and surface interactions were observed. Our results do not raise direct reason for concern with respect to crew health, but indicate a potential threat towards material integrity in moist areas. The International Space Station is a unique habitat for humans and microbes. Here, Mora et al. analyze microbial communities from several areas aboard, finding similarities with those of ground-based indoor environments, as well as adaptations towards biofilm formation but not necessarily relevant to human health.

Gravity waves (GWs) are among the key drivers of the meridional overturning circulation in the mesosphere and upper stratosphere. Their representation in climate models suffers from insufficient resolution and limited observational constraints on their parameterizations. This obscures assessments of middle atmospheric circulation changes in a changing climate. This study presents a comprehensive analysis of stratospheric GW activity above and downstream of the Andes from 1 to 15 August 2019, with special focus on GW representation ranging from an unprecedented kilometer-scale global forecast model (1.4 km ECMWF IFS), ground-based Rayleigh lidar (CORAL) observations, modern reanalysis (ERA5), to a coarse-resolution climate model (EMAC). Resolved vertical flux of zonal GW momentum (GWMF) is found to be stronger by a factor of at least 2–2.5 in IFS compared to ERA5. Compared to resolved GWMF in IFS, parameterizations in ERA5 and EMAC continue to inaccurately generate excessive GWMF poleward of 60°S, yielding prominent differences between resolved and parameterized GWMFs. A like-to-like validation of GW profiles in IFS and ERA5 reveals similar wave structures. Still, even at ∼1 km resolution, the resolved waves in IFS are weaker than those observed by lidar. Further, GWMF estimates across datasets reveal that temperature-based proxies, based on midfrequency approximations for linear GWs, overestimate GWMF due to simplifications and uncertainties in GW wavelength estimation from data. Overall, the analysis provides GWMF benchmarks for parameterization validation and calls for three-dimensional GW parameterizations, better upper-boundary treatment, and vertical resolution increases commensurate with increases in horizontal resolution in models, for a more realistic GW analysis.

Comprehensive studies of hydrogen embrittlement in high-strength austenitic alloys under cryogenic conditions are scarce, leaving the combined effect of hydrogen charging and extreme temperatures largely unexplored. Given the demands of cryogenic applications such as hydrogen storage and transport, understanding material behavior under these conditions is crucial. Here, we present the first systematic study of hydrogen’s effect at liquid helium temperature (4.2 K) on the mechanical properties of precipitation hardened austenitic alloys, specifically the nickel-based Alloy 718 and austenitic stainless steel A286. Both materials were subjected to pressurized hydrogen charging at 473 K followed by slow strain rate tensile testing at room temperature and at 4.2 K. Hydrogen charging caused significant ductility loss at room temperature in both alloys. In contrast, testing at 4.2 K resulted in increased strength and no evidence of hydrogen embrittlement. Notably, materials pre-charged with hydrogen and tested at 4.2 K exhibited higher stress drop amplitudes and increased strain accumulation during serration events, suggesting persistent hydrogen–dislocation interactions and possible enhanced dislocation pinning by obstacles such as Lomer–Cottrell locks. These results indicate that while hydrogen influences plasticity mechanisms at cryogenic temperatures, embrittlement is suppressed, providing new insight into the safe development of austenitic alloys in cryogenic hydrogen environments.