Cell Type-Specific Intralocus Interactions Reveal Oligodendrocyte Mechanisms in MS.

Multiple sclerosis (MS) is an autoimmune disease characterized by attack on oligodendrocytes within the central nervous system (CNS). Despite widespread use of immunomodulatory therapies, patients may still face progressive disability because of failure of myelin regeneration and loss of neurons, suggesting additional cellular pathologies. Here, we describe a general approach for identifying specific cell types in which a disease allele exerts a pathogenic effect. Applying this approach to MS risk loci, we pinpoint likely pathogenic cell types for 70%. In addition to T cell loci, we unexpectedly identified myeloid- and CNS-specific risk loci, including two sites that dysregulate transcriptional pause release in oligodendrocytes. Functional studies demonstrated inhibition of transcriptional elongation is a dominant pathway blocking oligodendrocyte maturation. Furthermore, pause release factors are frequently dysregulated in MS brain tissue. These data implicate cell-intrinsic aberrations outside of the immune system and suggest new avenues for therapeutic development. VIDEO ABSTRACT.

Drug-based modulation of endogenous stem cells promotes functional remyelination in vivo.

Multiple sclerosis involves an aberrant autoimmune response and progressive failure of remyelination in the central nervous system. Prevention of neural degeneration and subsequent disability requires remyelination through the generation of new oligodendrocytes, but current treatments exclusively target the immune system. Oligodendrocyte progenitor cells are stem cells in the central nervous system and the principal source of myelinating oligodendrocytes. These cells are abundant in demyelinated regions of patients with multiple sclerosis, yet fail to differentiate, thereby representing a cellular target for pharmacological intervention. To discover therapeutic compounds for enhancing myelination from endogenous oligodendrocyte progenitor cells, we screened a library of bioactive small molecules on mouse pluripotent epiblast stem-cell-derived oligodendrocyte progenitor cells. Here we show seven drugs function at nanomolar doses selectively to enhance the generation of mature oligodendrocytes from progenitor cells in vitro. Two drugs, miconazole and clobetasol, are effective in promoting precocious myelination in organotypic cerebellar slice cultures, and in vivo in early postnatal mouse pups. Systemic delivery of each of the two drugs significantly increases the number of new oligodendrocytes and enhances remyelination in a lysolecithin-induced mouse model of focal demyelination. Administering each of the two drugs at the peak of disease in an experimental autoimmune encephalomyelitis mouse model of chronic progressive multiple sclerosis results in striking reversal of disease severity. Immune response assays show that miconazole functions directly as a remyelinating drug with no effect on the immune system, whereas clobetasol is a potent immunosuppressant as well as a remyelinating agent. Mechanistic studies show that miconazole and clobetasol function in oligodendrocyte progenitor cells through mitogen-activated protein kinase and glucocorticoid receptor signalling, respectively. Furthermore, both drugs enhance the generation of human oligodendrocytes from human oligodendrocyte progenitor cells in vitro. Collectively, our results provide a rationale for testing miconazole and clobetasol, or structurally modified derivatives, to enhance remyelination in patients.

Modeling the Mutational and Phenotypic Landscapes of Pelizaeus-Merzbacher Disease with Human iPSC-Derived Oligodendrocytes.

Pelizaeus-Merzbacher disease (PMD) is a pediatric disease of myelin in the central nervous system and manifests with a wide spectrum of clinical severities. Although PMD is a rare monogenic disease, hundreds of mutations in the X-linked myelin gene proteolipid protein 1 (PLP1) have been identified in humans. Attempts to identify a common pathogenic process underlying PMD have been complicated by an incomplete understanding of PLP1 dysfunction and limited access to primary human oligodendrocytes. To address this, we generated panels of human induced pluripotent stem cells (hiPSCs) and hiPSC-derived oligodendrocytes from 12 individuals with mutations spanning the genetic and clinical diversity of PMD-including point mutations and duplication, triplication, and deletion of PLP1-and developed an in vitro platform for molecular and cellular characterization of all 12 mutations simultaneously. We identified individual and shared defects in PLP1 mRNA expression and splicing, oligodendrocyte progenitor development, and oligodendrocyte morphology and capacity for myelination. These observations enabled classification of PMD subgroups by cell-intrinsic phenotypes and identified a subset of mutations for targeted testing of small-molecule modulators of the endoplasmic reticulum stress response, which improved both morphologic and myelination defects. Collectively, these data provide insights into the pathogeneses of a variety of PLP1 mutations and suggest that disparate etiologies of PMD could require specific treatment approaches for subsets of individuals. More broadly, this study demonstrates the versatility of a hiPSC-based panel spanning the mutational heterogeneity within a single disease and establishes a widely applicable platform for genotype-phenotype correlation and drug screening in any human myelin disorder.

Colloidal crystal order and structure revealed by tabletop extreme ultraviolet scattering and coherent diffractive imaging.

Colloidal crystals with specific electronic, optical, magnetic, vibrational properties, can be rationally designed by controlling fundamental parameters such as chemical composition, scale, periodicity and lattice symmetry. In particular, silica nanospheres -which assemble to form colloidal crystals- are ideal for this purpose, because of the ability to infiltrate their templates with semiconductors or metals. However characterization of these crystals is often limited to techniques such as grazing incidence small-angle scattering that provide only global structural information and also often require synchrotron sources. Here we demonstrate small-angle Bragg scattering from nanostructured materials using a tabletop-scale setup based on high-harmonic generation, to reveal important information about the local order of nanosphere grains, separated by grain boundaries and discontinuities. We also apply full-field quantitative ptychographic imaging to visualize the extended structure of a silica close-packed nanosphere multilayer, with thickness information encoded in the phase. These combined techniques allow us to simultaneously characterize the silica nanospheres size, their symmetry and distribution within single colloidal crystal grains, the local arrangement of nearest-neighbor grains, as well as to quantitatively determine the number of layers within the sample. Key to this advance is the good match between the high harmonic wavelength used (13.5nm) and the high transmission, high scattering efficiency, and low sample damage of the silica colloidal crystal at this wavelength. As a result, the relevant distances in the sample - namely, the interparticle distance (≈124nm) and the colloidal grains local arrangement (≈1µm) - can be investigated with Bragg coherent EUV scatterometry and ptychographic imaging within the same experiment simply by tuning the EUV spot size at the sample plane (5µm and 15µm respectively). In addition, the high spatial coherence of high harmonics light, combined with advances in imaging techniques, makes it possible to image near-periodic structures quantitatively and nondestructively, and enables the observation of the extended order of quasi-periodic colloidal crystals, with a spatial resolution better than 20nm. In the future, by harnessing the high time-resolution of tabletop high harmonics, this technique can be extended to dynamically image the three-dimensional electronic, magnetic, and transport properties of functional nanosystems.

Underexplored biodiversity of Eastern Mediterranean biota: systematics and evolutionary history of the genus Aubrieta (Brassicaceae).

BACKGROUND AND AIMS: Aubrieta is a taxonomically difficult genus from the Brassicaceae family with approximately 20 species centred in Turkey and Greece. Species boundaries and their evolutionary history are poorly understood. Therefore, we analysed bio- and phylogeographic relationships and evaluated morphological variation to study the evolution of this genus. METHODS: Phylogenetic analyses of DNA sequence variation of nuclear-encoded loci and plastid DNA were used to unravel phylogeographic patterns. Morphometric analyses were conducted to study species delimitation. DNA sequence-based mismatch distribution and climate-niche analyses were performed to explain various radiations in space and time during the last 2·5 million years. KEY RESULTS: Species groups largely show non-overlapping distribution patterns in the eastern Mediterranean and Asia Minor. We recognized 20 species and provide evidence for overlooked species, thereby highlighting taxonomical difficulties but also demonstrating underexplored species diversity. The centre of origin of Aubrieta is probably Turkey, from which various clades expanded independently towards Asia Minor, south to Lebanon and west to Greece and the Balkans during the Pleistocene. CONCLUSIONS: Pleistocene climatic fluctuations had a pronounced effect on Aubrieta speciation and radiation during the last 1·1 million years in the Eastern Mediterranean and Asia Minor. In contrast to many other Brassicaceae, speciation processes did not involve excessive formation of polyploids, but displayed formation of diploids with non-overlapping present-day distribution areas. Expansions from the Aubrieta centre of origin and primary centre of species diversity showed adaptation trends towards higher temperature and drier conditions. However, later expansion and diversification of taxa from within the second centre of species diversity in Greece started ∼0·19 Mya and were associated with a general transition of species adaptation towards milder temperatures and less dry conditions.

Quantitative Chemically Specific Coherent Diffractive Imaging of Reactions at Buried Interfaces with Few Nanometer Precision.

We demonstrate quantitative, chemically specific imaging of buried nanostructures, including oxidation and diffusion reactions at buried interfaces, using nondestructive tabletop extreme ultraviolet (EUV) coherent diffractive imaging (CDI). Copper nanostructures inlaid in SiO2 are coated with 100 nm of aluminum, which is opaque to visible light and thick enough that neither visible microscopy nor atomic force microscopy can image the buried interface. Short wavelength high harmonic beams can penetrate the aluminum layer, yielding high-contrast images of the buried structures. Quantitative analysis shows that the reflected EUV light is extremely sensitive to the formation of multiple oxide layers, as well as interdiffusion of materials occurring at the metal-metal and metal-insulator boundaries deep within the nanostructure with few nanometers precision.

Ptychographic hyperspectral spectromicroscopy with an extreme ultraviolet high harmonic comb.

We report a proof-of-principle demonstration of a new scheme of spectromicroscopy in the extreme ultraviolet (EUV) spectral range, where the spectral response of the sample at different wavelengths is imaged simultaneously. This scheme is enabled by combining ptychographic information multiplexing (PIM) with a tabletop EUV source based on high harmonic generation, where four spectrally narrow harmonics near 30 nm form a spectral comb structure. Extending PIM from previously demonstrated visible wavelengths to the EUV/X-ray wavelengths promises much higher spatial resolution and a more powerful spectral contrast mechanism, making PIM an attractive spectromicroscopy method in both microscopy and spectroscopy aspects. In addition to spectromicroscopy, this method images the multicolor EUV beam in situ, making this a powerful beam characterization technique. In contrast to other methods, the techniques described here use no hardware to separate wavelengths, leading to efficient use of the EUV radiation.

Spatial, spectral, and polarization multiplexed ptychography.

We introduce a novel coherent diffraction imaging technique based on ptychography that enables simultaneous full-field imaging of multiple, spatially separate, sample locations. This technique only requires that diffracted light from spatially separated sample sites be mutually incoherent at the detector, which can be achieved using multiple probes that are separated either by wavelength or by orthogonal polarization states. This approach enables spatially resolved polarization spectroscopy from a single ptychography scan, as well as allowing a larger field of view to be imaged without loss in spatial resolution. Further, we compare the numerical efficiency of the multi-mode ptychography algorithm with a single mode algorithm.

Charge-induced equilibrium dynamics and structure at the Ag(001)-electrolyte interface.

The applied potential dependent rate of atomic step motion of the Ag(001) surface in weak NaF electrolyte has been measured using a new extension of the technique of X-ray Photon Correlation Spectroscopy (XPCS). For applied potentials between hydrogen evolution and oxidation, the surface configuration completely changes on timescales of 10(2)-10(4) seconds depending upon the applied potential. These dynamics, directly measured over large areas of the sample surface simultaneously, are related to the surface energy relative to over or under potential. Concurrent specular X-ray scattering measurements reveal how the ordering of the water layers at the interface correlates with the dynamics.

A world-wide perspective on crucifer speciation and evolution: phylogenetics, biogeography and trait evolution in tribe Arabideae.

BACKGROUND AND AIMS: Tribe Arabideae are the most species-rich monophyletic lineage in Brassicaceae. More than 500 species are distributed in the majority of mountain and alpine regions worldwide. This study provides the first comprehensive phylogenetic analysis for the species assemblage and tests for association of trait and characters, providing the first explanations for the enormous species radiation since the mid Miocene. METHODS: Phylogenetic analyses of DNA sequence variation of nuclear encoded loci and plastid DNA are used to unravel a reliable phylogenetic tree. Trait and ancestral area reconstructions were performed and lineage-specific diversification rates were calculated to explain various radiations in the last 15 Myr in space and time. KEY RESULTS: A well-resolved phylogenetic tree demonstrates the paraphyly of the genus Arabis and a new systematic concept is established. Initially, multiple radiations involved a split between lowland annuals and mountain/alpine perennial sister species. Subsequently, increased speciation rates occur in the perennial lineages. The centre of origin of tribe Arabideae is most likely the Irano-Turanian region from which the various clades colonized the temperate mountain and alpine regions of the world. CONCLUSIONS: Mid Miocene early diversification started with increased speciation rates due to the emergence of various annual lineages. Subsequent radiations were mostly driven by diversification within perennial species during the Pliocene, but increased speciation rates also occurred during that epoch. Taxonomic concepts in Arabis are still in need of a major taxonomic revision to define monophyletic groups.

Nuances in the effect of types of intimate partner violence on aspects of child development: Evidence from the Philippines.

BACKGROUND: Most studies on the effect of intimate partner violence (IPV) do not distinguish between the effects of the different types of IPVs on the various aspects of child development. OBJECTIVE: This study explores the effect of multiple dimensions of IPV, specifically physical violence, controlling behavior, and emotional abuse, on children's physical and cognitive development. PARTICIPANTS AND SETTING: The study uses data from the Cebu Longitudinal Health and Nutritional Survey (CLHNS). The study uses a final sample of n = 1506 index children in Cebu, Philippines. METHODS: We use information pertaining to: (1) presence of IPV in the household, (2) demographic characteristics, (3) mother's status and autonomy, (4) anthropometric data, and (5) children's scores on mathematics, English, language, and nonverbal reasoning tests. We employ a Propensity Score Matching (PSM) technique to analyze the effect of IPV on child development outcomes, conditional on observed characteristics. RESULTS: Maternal exposure to physical violence significantly reduces child's body mass index (BMI) (ATE: -0.344, p ≤ 0.05). On the other hand, mother's vulnerability to controlling behavior decreases children's test scores in mathematics (ATE: -3.346, p ≤ 0.01), English (ATE: -2.289, p ≤ 0.01), and nonverbal reasoning (ATE: -2.103, p ≤ 0.01). We do not find a significant link between emotional abuse and height, body mass index, and test scores. CONCLUSIONS: Our study shows that the deleterious effect of IPV on child development is IPV-specific. Exposure to physical violence tends to affect children's physical development while cognitive abilities are negatively associated with exposure to controlling behavior. These results suggest different types of ex-post interventions on both children and mothers to ensure their physical and mental well-being.

Three-dimensional topological magnetic monopoles and their interactions in a ferromagnetic meta-lattice.

Topological magnetic monopoles (TMMs), also known as hedgehogs or Bloch points, are three-dimensional (3D) non-local spin textures that are robust to thermal and quantum fluctuations due to the topology protection1-4. Although TMMs have been observed in skyrmion lattices1,5, spinor Bose-Einstein condensates6,7, chiral magnets8, vortex rings2,9 and vortex cores10, it has been difficult to directly measure the 3D magnetization vector field of TMMs and probe their interactions at the nanoscale. Here we report the creation of 138 stable TMMs at the specific sites of a ferromagnetic meta-lattice at room temperature. We further develop soft X-ray vector ptycho-tomography to determine the magnetization vector and emergent magnetic field of the TMMs with a 3D spatial resolution of 10 nm. This spatial resolution is comparable to the magnetic exchange length of transition metals11, enabling us to probe monopole-monopole interactions. We find that the TMM and anti-TMM pairs are separated by 18.3 ± 1.6 nm, while the TMM and TMM, and anti-TMM and anti-TMM pairs are stabilized at comparatively longer distances of 36.1 ± 2.4 nm and 43.1 ± 2.0 nm, respectively. We also observe virtual TMMs created by magnetic voids in the meta-lattice. This work demonstrates that ferromagnetic meta-lattices could be used as a platform to create and investigate the interactions and dynamics of TMMs. Furthermore, we expect that soft X-ray vector ptycho-tomography can be broadly applied to quantitatively image 3D vector fields in magnetic and anisotropic materials at the nanoscale.

Systematics and evolution of Arctic-Alpine Arabis alpina (Brassicaceae) and its closest relatives in the eastern Mediterranean.

PREMISE OF THE STUDY: The high mountains in southern Anatolia and the eastern Mediterranean are assumed to play a major role as a primary center of genetic diversity and species richness in Eurasia. We tested this hypothesis by focusing on the widespread perennial arctic-alpine Arabis alpina and its sympatrically distributed closest relatives in the eastern Mediterranean. METHODS: Plastid (trnL intron, trnL-F intergenic spacer) and nuclear (ITS) DNA sequence analysis was used for phylogenetic reconstruction. Broad-scale plastid haplotype analyses were conducted to infer ancestral biogeographic patterns. KEY RESULTS: Five Arabis species, identified from the eastern Mediterranean (Turkey mainland and Cyprus), evolved directly and independently from A. alpina, leaving Arabis alpina as a paraphyletic taxon. These species are not affected by hybridization or introgression, and species divergence took place at the diploid level during the Pleistocene. CONCLUSIONS: Pleistocene climate fluctuations produced local altitudinal range-shifts among mountain glacial survival areas, resulting not only in the accumulation of intraspecific genotype diversity but also in the formation of five local species. We also show that the closest sister group of Arabis alpina consists exclusively of annuals/winter annuals and diverged prior to Pleistocene climatic fluctuations during the colonization of the lowland Mediterranean landscape. These findings highlight that Anatolia is not only a center of species richness but also a center for life-history diversification.

Survey data on energy and fuel use of firms in economic zones in the Philippines.

The data describe characteristics, operations, utilities, and fuels used in the production of 115 manufacturing and agro-industrial firms in Philippine special economic zones. The data include information on the firm's production, sales, and schedules; electricity sources, requirements, and uses; the importance of various conventional fuels, and the firms' fuel expenditure in their major production processes. The data also include their employee's aptitude, knowledge, considerations, and opinions on alternative fuels and primary energies, and experiences in using them. The data were gathered through a series of focus group discussions (FGDs) in June 2019 and an online survey conducted in August to September 2019. The data can be used in the analysis of energy consumption and expenditure of manufacturing and agro-industrial firms in the Philippines. The respondents' knowledge of and perceptions toward adopting alternative fuels in their firms' production processes are useful in the analysis of future energy demand.

Nondestructive, high-resolution, chemically specific 3D nanostructure characterization using phase-sensitive EUV imaging reflectometry.

Next-generation nano- and quantum devices have increasingly complex 3D structure. As the dimensions of these devices shrink to the nanoscale, their performance is often governed by interface quality or precise chemical or dopant composition. Here, we present the first phase-sensitive extreme ultraviolet imaging reflectometer. It combines the excellent phase stability of coherent high-harmonic sources, the unique chemical sensitivity of extreme ultraviolet reflectometry, and state-of-the-art ptychography imaging algorithms. This tabletop microscope can nondestructively probe surface topography, layer thicknesses, and interface quality, as well as dopant concentrations and profiles. High-fidelity imaging was achieved by implementing variable-angle ptychographic imaging, by using total variation regularization to mitigate noise and artifacts in the reconstructed image, and by using a high-brightness, high-harmonic source with excellent intensity and wavefront stability. We validate our measurements through multiscale, multimodal imaging to show that this technique has unique advantages compared with other techniques based on electron and scanning probe microscopies.

Multiple beam ptychography for large field-of-view, high throughput, quantitative phase contrast imaging.

The ability to record large field-of-view images without a loss in spatial resolution is of crucial importance for imaging science. For most imaging techniques however, an increase in field-of-view comes at the cost of decreased resolution. Here we present a novel extension to ptychographic coherent diffractive imaging that permits simultaneous full-field imaging of multiple locations by illuminating the sample with spatially separated, interfering probes. This technique allows for large field-of-view imaging in amplitude and phase while maintaining diffraction-limited resolution, without an increase in collected data i.e. diffraction patterns acquired.

Full-field imaging of thermal and acoustic dynamics in an individual nanostructure using tabletop high harmonic beams.

Imaging charge, spin, and energy flow in materials is a current grand challenge that is relevant to a host of nanoenhanced systems, including thermoelectric, photovoltaic, electronic, and spin devices. Ultrafast coherent x-ray sources enable functional imaging on nanometer length and femtosecond timescales particularly when combined with advances in coherent imaging techniques. Here, we combine ptychographic coherent diffractive imaging with an extreme ultraviolet high harmonic light source to directly visualize the complex thermal and acoustic response of an individual nanoscale antenna after impulsive heating by a femtosecond laser. We directly image the deformations induced in both the nickel tapered nanoantenna and the silicon substrate and see the lowest-order generalized Lamb wave that is partially confined to a uniform nanoantenna. The resolution achieved-sub-100 nm transverse and 0.5-Å axial spatial resolution, combined with ≈10-fs temporal resolution-represents a significant advance in full-field dynamic imaging capabilities. The tapered nanoantenna is sufficiently complex that a full simulation of the dynamic response would require enormous computational power. We therefore use our data to benchmark approximate models and achieve excellent agreement between theory and experiment. In the future, this work will enable three-dimensional functional imaging of opaque materials and nanostructures that are sufficiently complex that their functional properties cannot be predicted.

Occurrence and source identification of perfluoroalkyl acids (PFAAs) in the Metedeconk River Watershed, New Jersey.

The Brick Township Municipal Utilities Authority (BTMUA), which relies on the Metedeconk River as its primary source of water supply, initiated a perfluoroalkyl acid (PFAA) source trackdown study in collaboration with the New Jersey Department of Environmental Protection (NJDEP) after discovering that the concentration of one PFAA, perfluorooctanoic acid (PFOA), was elevated at their raw surface water intake. Water samples were collected over eight sampling events between September 2011 and July 2014. Samples included surface water, groundwater, stormwater, sanitary sewer water, and commercial/industrial process water. Each sample was analyzed for ten PFAAs. Results from a set of samples collected from the 80 km2 South Branch Metedeconk River watershed directed the focus of this study to a 7.5-km2 area of interest. Within this area, a high concentration of PFAA contamination was documented in a localized zone. Subsequent groundwater sampling led to the identification of a plume of groundwater contamination emanating from an industrial/business park. The suspected source of PFAA detected in the river and drinking water intake was identified to a small industrial facility that used materials containing PFOA. Groundwater PFOA concentrations as high as 70,000 ng/L were found in samples taken within 200 m of the parcel and surface water concentrations as high as 130 ng/L were observed in the river. While various PFAAs were detected in the samples, particularly in groundwater samples, PFOA was identified as the primary contaminant of concern with respect to the river and the BTMUA water supply.