Mechanisms of manganese(II) oxidation by filamentous ascomycete fungi vary with species and time as a function of secretome composition

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Zeiner, C. A., Purvine, S. O., Zink, E., Wu, S., Pasa-Tolic, L., Chaput, D. L., Santelli, C. M., & Hansel, C. M. Mechanisms of manganese(II) oxidation by filamentous ascomycete fungi vary with species and time as a function of secretome composition. Frontiers in Microbiology, 12, (2021): 610497, https://doi.org/10.3389/fmicb.2021.610497.Manganese (Mn) oxides are among the strongest oxidants and sorbents in the environment, and Mn(II) oxidation to Mn(III/IV) (hydr)oxides includes both abiotic and microbially-mediated processes. While white-rot Basidiomycete fungi oxidize Mn(II) using laccases and manganese peroxidases in association with lignocellulose degradation, the mechanisms by which filamentous Ascomycete fungi oxidize Mn(II) and a physiological role for Mn(II) oxidation in these organisms remain poorly understood. Here we use a combination of chemical and in-gel assays and bulk mass spectrometry to demonstrate secretome-based Mn(II) oxidation in three phylogenetically diverse Ascomycetes that is mechanistically distinct from hyphal-associated Mn(II) oxidation on solid substrates. We show that Mn(II) oxidative capacity of these fungi is dictated by species-specific secreted enzymes and varies with secretome age, and we reveal the presence of both Cu-based and FAD-based Mn(II) oxidation mechanisms in all 3 species, demonstrating mechanistic redundancy. Specifically, we identify candidate Mn(II)-oxidizing enzymes as tyrosinase and glyoxal oxidase in Stagonospora sp. SRC1lsM3a, bilirubin oxidase in Stagonospora sp. and Paraconiothyrium sporulosum AP3s5-JAC2a, and GMC oxidoreductase in all 3 species, including Pyrenochaeta sp. DS3sAY3a. The diversity of the candidate Mn(II)-oxidizing enzymes identified in this study suggests that the ability of fungal secretomes to oxidize Mn(II) may be more widespread than previously thought.This work was supported by the National Science Foundation, grant numbers EAR-1249489 and CBET-1336496, both awarded to CH, by a JGI-EMSL Collaborative Science Initiative grant (proposal number 48100) awarded to CH and CS, and by the University of St. Thomas. Personal support for CZ was also provided by Harvard University and by a Ford Foundation Predoctoral Fellowship administered by the National Academies. A portion of this research was performed under the Facilities Integrating Collaborations for User Science (FICUS) program and used resources at the DOE Joint Genome Institute and the Environmental Molecular Sciences Laboratory (grid.436923.9), which are DOE Office of Science User Facilities. Both facilities are sponsored by the Biological and Environmental Research Program and operated under Contract Nos. DE-AC02-05CH11231 (JGI) and DE-AC05-76RL01830 (EMSL). Part of this research was performed at the Bauer Core Facility of the FAS Center for Systems Biology at Harvard University. A portion of the bioinformatics analysis was performed at Harvard’s FAS Research Computing facility

Validation of monoclonal antibody F99/97.6.1 for immunohistochemical staining of brain and tonsil in mule deer (\u3ci\u3eOdocoileus hemionus\u3c/i\u3e) with chronic wasting disease

A new monoclonal antibody (MAb), F99/97.6.1, that has been used to demonstrate scrapieassociated prion protein PrPSc in brain and lymphoid tissues of domestic sheep with scrapie was used in an immunohistochemistry assay for diagnosis of chronic wasting disease (CWD) in mule deer (Odocoileus hemionus). The MAb F99/97.6.1 immunohistochemistry assay was evaluated in brain and tonsil tissue from 100 mule deer that had spongiform encephalopathy compatible with CWD and from 1,050 mule deer outside the CWD-endemic area. This MAb demonstrated abnormal protease-resistant prion protein (PrPres) in brains of all of the 100 mule deer and in 99 of the 100 tonsil samples. No immunostaining was seen in samples collected from deer outside the endemic area. MAb F99/97.6.1 demonstrated excellent properties for detection of PrPres in fresh, frozen, or mildly to moderately autolytic samples of brain and tonsil. This immunohistochemistry assay is a sensitive, specific, readily standardized diagnostic test for CWD in deer

Validation of monoclonal antibody F99/97.6.1 for immunohistochemical staining of brain and tonsil in mule deer (\u3ci\u3eOdocoileus hemionus\u3c/i\u3e) with chronic wasting disease

A new monoclonal antibody (MAb), F99/97.6.1, that has been used to demonstrate scrapieassociated prion protein PrPSc in brain and lymphoid tissues of domestic sheep with scrapie was used in an immunohistochemistry assay for diagnosis of chronic wasting disease (CWD) in mule deer (Odocoileus hemionus). The MAb F99/97.6.1 immunohistochemistry assay was evaluated in brain and tonsil tissue from 100 mule deer that had spongiform encephalopathy compatible with CWD and from 1,050 mule deer outside the CWD-endemic area. This MAb demonstrated abnormal protease-resistant prion protein (PrPres) in brains of all of the 100 mule deer and in 99 of the 100 tonsil samples. No immunostaining was seen in samples collected from deer outside the endemic area. MAb F99/97.6.1 demonstrated excellent properties for detection of PrPres in fresh, frozen, or mildly to moderately autolytic samples of brain and tonsil. This immunohistochemistry assay is a sensitive, specific, readily standardized diagnostic test for CWD in deer

Experimental investigation of homogeneous freezing of sulphuric acid particles in the aerosol chamber AIDA

The homogeneous freezing of supercooled H₂SO₄/H₂O solution droplets was investigated in the aerosol chamber AIDA (Aerosol Interactions and Dynamics in the Atmosphere) of Forschungszentrum Karlsruhe. 24 freezing experiments were performed at temperatures between 189 and 235 K with aerosol particles in the diameter range 0.05 to 1 µm. Individual experiments started at homogeneous temperatures and ice saturation ratios between 0.9 and 0.95. Cloud cooling rates up to -2.8 K min^-1 were simulated dynamically in the chamber by expansion cooling using a mechanical pump. Depending on the cooling rate and starting temperature, freezing threshold relative humidities were exceeded after expansion time periods between about 1 and 10 min. The onset of ice formation was measured with three independent methods showing good agreement among each other. Ice saturation ratios measured at the onset of ice formation increased from about 1.4 at 231 K  to about 1.75 at 189 K. The experimental data set including thermodynamic parameters as well as physical and chemical aerosol analysis provides a good basis for microphysical model applications

Dynamics of Weak First Order Phase Transitions

The dynamics of weak vs. strong first order phase transitions is investigated numerically for 2+1 dimensional scalar field models. It is argued that the change from a weak to a strong transition is itself a (second order) phase transition, with the order parameter being the equilibrium fractional population difference between the two phases at the critical temperature, and the control parameter being the coefficient of the cubic coupling in the free-energy density. The critical point is identified, and a power law controlling the relaxation dynamics at this point is obtained. Possible applications are briefly discussed.Comment: 11 pages, 4 figures in uuencoded compressed file (see instructions in main text), RevTeX, DART-HEP-94/0

Stability of general-relativistic accretion disks

Self-gravitating relativistic disks around black holes can form as transient structures in a number of astrophysical scenarios such as binary neutron star and black hole-neutron star coalescences, as well as the core-collapse of massive stars. We explore the stability of such disks against runaway and non-axisymmetric instabilities using three-dimensional hydrodynamics simulations in full general relativity using the THOR code. We model the disk matter using the ideal fluid approximation with a $\Gamma$-law equation of state with $\Gamma=4/3$. We explore three disk models around non-rotating black holes with disk-to-black hole mass ratios of 0.24, 0.17 and 0.11. Due to metric blending in our initial data, all of our initial models contain an initial axisymmetric perturbation which induces radial disk oscillations. Despite these oscillations, our models do not develop the runaway instability during the first several orbital periods. Instead, all of the models develop unstable non-axisymmetric modes on a dynamical timescale. We observe two distinct types of instabilities: the Papaloizou-Pringle and the so-called intermediate type instabilities. The development of the non-axisymmetric mode with azimuthal number m = 1 is accompanied by an outspiraling motion of the black hole, which significantly amplifies the growth rate of the m = 1 mode in some cases. Overall, our simulations show that the properties of the unstable non-axisymmetric modes in our disk models are qualitatively similar to those in Newtonian theory.Comment: 30 pages, 21 figure

Thermodynamic properties of excess-oxygen-doped La2CuO4.11 near a simultaneous transition to superconductivity and long-range magnetic order

We have measured the specific heat and magnetization {\it versus} temperature in a single crystal sample of superconducting La$_{2}$CuO$_{4.11}$ and in a sample of the same material after removing the excess oxygen, in magnetic fields up to 15 T. Using the deoxygenated sample to subtract the phonon contribution, we find a broad peak in the specific heat, centered at 50 K. This excess specific heat is attributed to fluctuations of the Cu spins possibly enhanced by an interplay with the charge degrees of freedom, and appears to be independent of magnetic field, up to 15 T. Near the superconducting transition $T_{c}$($H$=0)= 43 K, we find a sharp feature that is strongly suppressed when the magnetic field is applied parallel to the crystallographic c-axis. A model for 3D vortex fluctuations is used to scale magnetization measured at several magnetic fields. When the magnetic field is applied perpendicular to the c-axis, the only observed effect is a slight shift in the superconducting transition temperature.Comment: 8 pages, 8 figure

A systematic review of methods used to study fish in saltmarsh flats

There is a growing body of research highlighting the importance of saltmarshes as habitats for fish for feeding, refuge from predation and reproduction. However, more work is needed on fish on vegetated marsh flats (or surfaces). We reviewed 60 studies that used 21 methods to sample fish assemblages on saltmarsh flats. Drop samplers, fyke nets and pop nets were most frequently employed, with considerably more studies being conducted in graminoid than succulent marsh. Reporting of sampling temporal and tidal details, environmental variables and fish attributes was inconsistent. Most of the papers focussed on one or more of conservation management, comparisons among habitat types, and the use of saltmarsh (including fish activity type or residency status). Important potential areas of research include the relationships between the fish assemblages of saltmarsh flats and coastal fisheries, the effects of invasive plant species and marsh restoration efforts in areas outside the United States, and the potential effects of sea-level rise on vegetated flats as fish habitat. Sampling methods that provide density measures are likely to be most useful for most of this research. Thus, drop samplers and pop nets are an appropriate choice, the former in graminoid saltmarshes and the latter in succulent saltmarshes

Ramification theory for varieties over a local field

We define generalizations of classical invariants of wild ramification for coverings on a variety of arbitrary dimension over a local field. For an l-adic sheaf, we define its Swan class as a 0-cycle class supported on the wild ramification locus. We prove a formula of Riemann-Roch type for the Swan conductor of cohomology together with its relative version, assuming that the local field is of mixed characteristic. We also prove the integrality of the Swan class for curves over a local field as a generalization of the Hasse-Arf theorem. We derive a proof of a conjecture of Serre on the Artin character for a group action with an isolated fixed point on a regular local ring, assuming the dimension is 2.Comment: 159 pages, some corrections are mad