Single Injection AAV2-FGF18 Gene Therapy Reduces Cartilage Loss and Subchondral Bone Damage in a Mechanically Induced Model of Osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a highly debilitating, degenerative pathology of cartilaginous joints affecting over 500 million people worldwide. The global economic burden of OA is estimated at $260-519 billion and growing, driven by aging global population and increasing rates of obesity. To date, only the multi-injection chondroanabolic treatment regimen of Fibroblast Growth Factor 18 (FGF18) has demonstrated clinically meaningful disease-modifying efficacy in placebo-controlled human trials. Our work focuses on the development of a novel single injection disease-modifying gene therapy, based on FGF18's chondroanabolic activity. METHODS: OA was induced in Sprague-Dawley rats using destabilization of the medial meniscus (DMM) (3 weeks), followed by intra-articular treatment with 3 dose levels of AAV2-FGF18, rh- FGF18 protein, and PBS. Durability, redosability, and biodistribution were measured by quantifying nLuc reporter bioluminescence. Transcriptomic analysis was performed by RNA-seq on cultured human chondrocytes and rat knee joints. Morphological analysis was performed on knee joints stained with Safranin O/Fast Green and anti-PRG antibody. RESULTS: Dose-dependent reductions in cartilage defect size were observed in the AAV2-FGF18- treated joints relative to the vehicle control. Total defect width was reduced by up to 76% and cartilage thickness in the thinnest zone was increased by up to 106%. Morphologically, the vehicle- treated joints exhibited pronounced degeneration, ranging from severe cartilage erosion and bone void formation, to subchondral bone remodeling and near-complete subchondral bone collapse. In contrast, AAV2-FGF18-treated joints appeared more anatomically normal, with only regional glycosaminoglycan loss and marginal cartilage erosion. While effective at reducing cartilage lesions, treatment with rhFGF18 injections resulted in significant joint swelling (19% increase in diameter), as well as a decrease in PRG4 staining uniformity and intensity. In contrast to early-timepoint in vitro RNA-seq analysis, which showed a high degree of concordance between protein- and gene therapy-treated chondrocytes, in vivo transcriptomic analysis, revealed few gene expression changes following protein treatment. On the other hand, the gene therapy treatment exhibited a high degree of durability and localization over the study period, upregulating several chondroanabolic genes while downregulating OA- and fibrocartilage-associated markers. CONCLUSION: FGF18 gene therapy treatment of OA joints can provide benefits to both cartilage and subchondral bone, with a high degree of localization and durability.

Development and characterization of a first-in-class adjustable-dose gene therapy system.

INTRODUCTION: Despite significant potential, gene therapy has been relegated to the treatment of rare diseases, due in part to an inability to adjust dosage following initial administration. Other significant constraints include cost, specificity, antigenicity, and systemic toxicity of current generation technologies. To overcome these challenges, we developed a first-in-class adjustable-dose gene therapy system, with optimized biocompatibility, localization, durability, and cost. METHODS: A lipid nanoparticle (LNP) delivery system was developed and characterized by dynamic light scattering for size, zeta potential, and polydispersity. Cytocompatibility and transfection efficiency were optimized in vitro using primary human adipocytes and preadipocytes. Durability, immunogenicity, and adjustment of expression were evaluated in C57BL/6 and B6 albino mice using in vivo bioluminescence imaging. Biodistribution was assessed by qPCR and immunohistochemistry; therapeutic protein expression was quantified by ELISA. RESULTS: Following LNP optimization, in vitro transfection efficiency of primary human adipocytes reached 81.3 % ± 8.3 % without compromising cytocompatibility. Critical physico-chemical properties of the system (size, zeta potential, polydispersity) remained stable over a broad range of genetic cassette sizes (1,871-6,203 bp). Durable expression was observed in vivo over 6 months, localizing to subcutaneous adipose tissues at the injection site with no detectable transgene in the liver, heart, spleen, or kidney. Gene expression was adjustable using several physical and pharmacological approaches, including cryolipolysis, focused ultrasound, and pharmacologically inducible apoptosis. The ability of transfected adipocytes to express therapeutic transgenes ranging from peptides to antibodies, at potentially clinically relevant levels, was confirmed in vitro and in vivo. CONCLUSION: We report the development of a novel, low-cost therapeutic platform, designed to enable the replacement of subcutaneously administered protein treatments with a single-injection, adjustable-dose gene therapy.

African Mountain Thistles: Three New Genera in the Carduus-Cirsium Group.

The floras on the highest mountains in tropical eastern Africa are among the most unique floras in the world. Despite the exceptionally high concentration of endemic species, these floras remain understudied from an evolutionary point of view. In this study, we focus on the Carduus-Cirsium group (subtribe Carduinae) to unravel the evolutionary relationships of the species endemic to the tropical Afromontane and Afroalpine floras, aiming to improve the systematics of the group. We applied the Hyb-Seq approach using the Compositae1061 probe set on 190 samples (159 species), encompassing representatives of all genera of Carduinae. We used two recently developed pipelines that enabled the processing of raw sequence reads, identification of paralogous sequences and segregation into orthologous alignments. After the implementation of a missing data filter, we retained sequences from 986 nuclear loci and 177 plastid regions. Phylogenomic analyses were conducted using both concatenated and summary-coalescence methods. The resulting phylogenies were highly resolved and revealed three distinct evolutionary lineages consisting of the African species traditionally referred to as Carduus and Cirsium. Consequently, we propose the three new genera Afrocarduus, Afrocirsium and Nuriaea; the latter did notably not belong to the Carduus-Cirsium group. We detected some incongruences between the phylogenies based on concatenation vs. coalescence and on nuclear vs. plastid datasets, likely attributable to incomplete lineage sorting and/or hybridization.

Transition to Self-compatibility Associated With Dominant S-allele in a Diploid Siberian Progenitor of Allotetraploid Arabidopsis kamchatica Revealed by Arabidopsis lyrata Genomes.

A transition to selfing can be beneficial when mating partners are scarce, for example, due to ploidy changes or at species range edges. Here, we explain how self-compatibility evolved in diploid Siberian Arabidopsis lyrata, and how it contributed to the establishment of allotetraploid Arabidopsis kamchatica. First, we provide chromosome-level genome assemblies for two self-fertilizing diploid A. lyrata accessions, one from North America and one from Siberia, including a fully assembled S-locus for the latter. We then propose a sequence of events leading to the loss of self-incompatibility in Siberian A. lyrata, date this independent transition to ∼90 Kya, and infer evolutionary relationships between Siberian and North American A. lyrata, showing an independent transition to selfing in Siberia. Finally, we provide evidence that this selfing Siberian A. lyrata lineage contributed to the formation of the allotetraploid A. kamchatica and propose that the selfing of the latter is mediated by the loss-of-function mutation in a dominant S-allele inherited from A. lyrata.

Adeno-Associated Virus-Delivered Fibroblast Growth Factor 18 Gene Therapy Promotes Cartilage Anabolism.

OBJECTIVE: To determine the characterization of chondrogenic properties of adeno-associated virus type 2 (AAV2)-delivered hFGF18, via analysis of effects on primary human chondrocyte proliferation, gene expression, and in vivo cartilage thickness changes in the tibia and meniscus. DESIGN: Chondrogenic properties of AAV2-FGF18 were compared with recombinant human FGF18 (rhFGF18) in vitro relative to phosphate-buffered saline (PBS) and AAV2-GFP negative controls. Transcriptome analysis was performed using RNA-seq on primary human chondrocytes treated with rhFGF18 and AAV2-FGF18, relative to PBS. Durability of gene expression was assessed using AAV2-nLuc and in vivo imaging. Chondrogenesis was evaluated by measuring weight-normalized thickness in the tibial plateau and the white zone of the anterior horn of the medial meniscus in Sprague-Dawley rats. RESULTS: AAV2-FGF18 elicits chondrogenesis by promoting proliferation and upregulation of hyaline cartilage-associated genes, including COL2A1 and HAS2, while downregulating fibrocartilage-associated COL1A1. This activity translates to statistically significant, dose-dependent increases in cartilage thickness in vivo within the area of the tibial plateau, following a single intra-articular injection of the AAV2-FGF18 or a regimen of 6 twice-weekly injections of rhFGF18 protein relative to AAV2-GFP. In addition, we observed AAV2-FGF18-induced and rhFGF18-induced increases in cartilage thickness of the anterior horn of the medial meniscus. Finally, the single-injection AAV2-delivered hFGF18 offers a potential safety advantage over the multi-injection protein treatment as evidenced by reduced joint swelling over the study period. CONCLUSION: AAV2-delivered hFGF18 represents a promising strategy for the restoration of hyaline cartilage by promoting extracellular matrix production, chondrocyte proliferation, and increasing articular and meniscal cartilage thickness in vivo after a single intra-articular injection.

From everywhere all at once: Several colonization routes available to Svalbard in the early Holocene.

For many arctic species, the spatial (re-)colonization patterns after the last Pleistocene glaciation have been described. However, the temporal aspects of their colonization are largely missing. Did one route prevail early, while another was more important later? The high Arctic archipelago Svalbard represents a good model system to address timeframe of postglacial plant colonization. Svalbard was almost fully glaciated during last glacial maximum and (re-)colonization of vascular plants began in early Holocene. Early Holocene climatic optimum (HCO) supported an expanded establishment of a partly thermophilic vegetation. Today, we find remnants of this vegetation in sheltered regions referred to as "Arctic biodiversity hotspots". The oldest record of postglacial plant colonization to Svalbard is found in Ringhorndalen-Flatøyrdalen. Even though thermophilic species could establish also later in Holocene, only HCO was favorable for vast colonization, and only hotspots offered stable conditions for thermophilic populations throughout Holocene. Thus, these relic populations may reflect colonization patterns of HCO. We investigate whether the colonization direction of thermophilic plants (Arnica angustifolia, Campanula uniflora, Pinguicula alpina, Tofieldia pusilla, and Vaccinium uliginosum ssp. microphyllum) in Ringhorndalen-Flatøyrdalen was uniform and different from later colonization events in other localities and non-thermophilic plants (Arenaria humifusa, Bistorta vivipara, Juncus biglumis, Oxyria digyna, and Silene acaulis). We analyzed plastid haplotypes of the 10 taxa from Ringhorndalen-Flatøyrdalen, from later-colonized localities in Svalbard, and from putative source regions outside Svalbard. Only rare and thermophilic taxa Campanula uniflora and Vaccinium uliginosum ssp. microphyllum provided results suggesting at least two colonization events from different source regions. Tofieldia pusilla and all the non-thermophilic plants showed no clear phylogeographically differentiation within Svalbard. Two of the thermophilic species showed no sequence variation. Based on the results, a uniform colonization direction to Svalbard in early Holocene is not probable; several source areas and dispersal directions were contemporarily involved.

Comparative evaluation of rhFGF18 and rhGDF11 treatment in a transient ischemia stroke model.

Background: Pharmacological treatments for ischemic stroke remain limited to thrombolysis, which is associated with increased risk of potentially fatal hemorrhage. Treatments with Recombinant Human Fibroblast Growth Factor 18 (rhFGF18) and Growth and Differentiation Factor 11 (rhGDF11) appear promising based on different preclinical models. The goal of this study was to compare the effects of rhFGF18 and rhGDF11 directly on survival, behavioral deficits, and histological fingerprint of cerebral ischemia in the Wistar rat middle cerebral artery occlusion (MCAO) model of stroke. Methods: Ischemia-reperfusion injury was induced using a 2-hour transient MCAO. Animals were administered rhFGF18 (infusion), rhGDF11 (multi-injection), or Phosphate Buffered Saline (PBS) vehicle control and followed for 42 days. Motor-Cognitive deficits were evaluated using the Morris Water Maze at Days 0 (pre-MCAO), 7, 21, and 42. Histopathological assessments were performed on Days 21 and 42. Results: Day 7 post-ischemia water maze performance times increased 38.3%, 2.1%, and 23.1% for PBS, rhFGF18, and rhGDF11-treated groups, respectively. Fraction of neurons with abnormal morphology (chromatolysis, pyknotic nuclei, somal degeneration) decreased in all groups toward Day 42 and was lowest for rhFGF18. AChE-positive fiber density and activity increased over time in the rhFGF18 group, remained unchanged in the rhGDF11 treatment arm, and declined in the PBS control. Metabolic increases were greatest in rhGDF11 treated animals, with both rhFGF18 and rhGDF11 achieving improvements over PBS, as evidenced by increased succinate dehydrogenase and lactate dehydrogenase activity. Finally, rhFGF18 treatment exhibited a trend for reduced mortality relative to PBS (5.6%, 95% CI [27.3%, 0.1% ] vs. 22.2%, 95% CI [47.6%, 6.4% ]). Conclusions: rhFGF18 treatment appears promising in improving survival and promoting motor-cognitive recovery following cerebral ischemia-reperfusion injury.

The benefits of contributing to the citizen science platform iNaturalist as an identifier.

As the number of observations submitted to the citizen science platform iNaturalist continues to grow, it is increasingly important that these observations can be identified to the finest taxonomic level, maximizing their value for biodiversity research. Here, we explore the benefits of acting as an identifier on iNaturalist.

Effect of Magnetic Impurities on Superconductivity in LaH10.

Polyhydrides are a novel class of superconducting materials with extremely high critical parameters, which is very promising for sensor applications. On the other hand, a complete experimental study of the best so far known superconductor, lanthanum superhydride LaH10 , encounters a serious complication because of the large upper critical magnetic field HC2 (0), exceeding 120-160 T. It is found that partial replacement of La atoms by magnetic Nd atoms results in significant suppression of superconductivity in LaH10 : each at% of Nd causes a decrease in TC by 10-11 K, helping to control the critical parameters of this compound. Strong pulsed magnetic fields up to 68 T are used to study the Hall effect, magnetoresistance, and the magnetic phase diagram of ternary metal polyhydrides for the first time. Surprisingly, (La,Nd)H10 demonstrates completely linear HC2 (T) âˆ |T - TC |, which calls into question the applicability of the Werthamer-Helfand-Hohenberg model for polyhydrides. The suppression of superconductivity in LaH10 by magnetic Nd atoms and the robustness of TC with respect to nonmagnetic impurities (e.g., Y, Al, C) under Anderson's theorem gives new experimental evidence of the isotropic (s-wave) character of conventional electron-phonon pairing in lanthanum decahydride.

MHA Herbarium: Collections of mosses from Yana-Indigirka Region, Yakutia, Russia.

BACKGROUND: The Skvortsov Herbarium of the Tsitsin Main Botanical Garden, Russian Academy of Sciences (MHA) in the 1945-1980s dealt with vascular plants and only scattered occasional collections of bryophytes and lichens were accumulated there without special arrangement. Since the late 1980s, the bryophyte studies in the MHA Herbarium became permanent and several projects were started since then, including the currently conducted "Moss Flora of Russia". There are many white spots on the map of bryophyte exploration of Russia, but one of the most conspicuous was Yakutia, the largest administrative unit of Russia, covering 3,081,000 km2. Yana-Indigirka Region, originally defined as a floristic region, includes Verkhoyansky Range and some smaller adjacent mountain areas. It is the largest amongst the bryofloristic regions in Russia, but exploration of its territory, which is difficult to access, remains far from complete. NEW INFORMATION: Several expeditions of the Institute for Biological Problems of Cryolithozone, Siberian Branch of Russian Academy of Sciences, and the Main Botanical Garden, Russian Academy of Sciences in 2000-2018 yielded in many bryophyte specimens, partly published in a number of papers. This dataset comprehensively represents the diversity of mosses of the Region. It includes 7,738 records of moss specimens preserved in the MHA Herbarium.

Fleroff goes digital: georeferenced records from "Flora des Gouvernements Wladimir" (Fleroff, 1902).

BACKGROUND: Global Biodiversity Information Facility (GBIF) has uneven data coverage across taxonomic, spatial and temporal dimensions. Temporal imbalances in the data coverage are particularly dramatic. Thus, 188.3M GBIF records were made in 2020, more than the whole lot of the currently available pre-1986 electronic data. This underscores the importance of reliable and precise biodiversity spatial data collected in early times. Biological collections certainly play a key role in our knowledge of biodiversity in the past. However, digitisation of historical literature is underway, being a modern trend in biodiversity data mining. The grid dataset for the flora of Vladimir Oblast, Russia, includes many historical records borrowed from the "Flora des Gouvernements Wladimir" by Alexander F. Fleroff (also known as Flerov or Flerow). Intensive study of Fleroff's collections and field surveys exactly in the same localities where he worked, showed that the quality of his data is superb. Species lists collected across hundreds of localities form a unique source of reliable information on the floristic diversity of Vladimir Oblast and adjacent areas for the period from 1894 to 1901. Since the grid dataset holds generalised data, we made precise georeferencing of Fleroff's literature records and published them in the form of a GBIF-mediated dataset. NEW INFORMATION: A dataset, based on "Flora des Gouvernements Wladimir. I. Pflanzengeographische Beschreibung des Gouvernements Wladimir" by Fleroff (1902), includes 8,889 records of 654 taxa (mainly species) from 366 localities. The majority of records originate from Vladimir Oblast (4,611 records of 534 taxa from 195 localities) and Yaroslavl Oblast (2,013 records of 409 taxa from 66 localities), but also from Nizhny Novgorod Oblast (942 records), Ivanovo Oblast (667 records) and Moscow Oblast (656 records). The leading second-level administrative units by the number of records are Pereslavsky District (2,013 records), Aleksandrovsky District (1,318 records) and Sergievo-Posadsky District (599 records). Georeferencing was carried out, based on the expert knowledge of the area, analysis of modern satellite images and old topographic maps. For 2,460 records, the georeferencing accuracy is 1,000 m or less (28%), whereas for 6,070 records it is 2,000 m or less (68%). The mean accuracy of records of the entire dataset is 2,447 m. That accuracy is unattainable for most herbarium collections of the late 19th century. Some localities of rare plants discovered by Fleroff and included into the dataset were completely lost in the 20th century due to either peat mining or development of urban areas.

A grid-based database on vascular plant distribution in the Meshchersky National Park, Ryazan Oblast, Russia.

BACKGROUND: Ryazan Oblast, situated in the central part of European Russia, has a long tradition of biodiversity research. Large distributional, ecological and phenological data on various taxonomic groups are available from this territory, mainly in the form of paper publications items, undigitised museum collections and archival sources. The purpose of this dataset is to deliver floristic materials, collected by the authors in the Meshchera Lowlands in the form of GBIF-mediated electronic data, to a wider audience. The dataset covers wild tracheophytes (native species, naturalised aliens and casuals) of the Meshchersky National Park. In 2020, it was used for the production of grid maps in "Flora of the Meschchersky National Park: checklist and atlas". NEW INFORMATION: The dataset contains 14,476 grid records of 817 taxa (806 species and hybrids, ten species aggregates and one genus). Most of the records (82.4%) were made in the field by A.V. Shcherbakov, M.V. Kazakova, N.V. Lyubeznova and A.D. Pastushenko in 2017 and 2018. The dataset includes only one occurrence per species per grid square. Georeferences are based on the WGS84 grid scheme with 55 squares measuring ca. 25 km2 (2.5' lat. × 5' long.). Each occurrence is linked to the corresponding grid square centroid; therefore, actual coordinates, habitat details and voucher information are unavailable. As of September 2021, the dataset on the flora of the Meshchersky National Park represents the second largest dataset on the biodiversity of Ryazan Oblast, Russia, published in GBIF.

Contribution to the "Atlas of the Russian Flora": Twelve local floras of European Russia.

BACKGROUND: The purpose of this dataset is to deliver to a wider audience in the form of GBIF-mediated data vast floristic materials collected by the author across various localities of European Russia from 2001-2019 (Arkhangelsk, Tver, Vladimir, Tula, Lipetsk, Voronezh Oblasts, Krasnodar Krai, City of Moscow and Komi Republic). Taxonomic data on vascular plants for ten locations were mobilised from the papers and technical reports published in Russian and standardised. Floristic treatments for two locations (Yasnaya Polyana and Tsaritsyno) have never been published before. NEW INFORMATION: The newly-prepared dataset includes 5,309 species records, i.e. one species record per each local flora. These are either native or alien (fully naturalised and casual) species. All records within one local flora have the same centroid coordinates and coordinate uncertainty in metres. Floristic inventories from the following locations were mobilised: 01. Ustya, Arkhangelsk Oblast (543 species, 1,500 km2); 02. Zaseki, Tula Oblast (593 species, 60 km2); 03. Polibino, Lipetsk Oblast (553 species, 70 km2); 04. Khrenovoye, Voronezh Oblast (665 species, 200 km2); 05. Troyeruchitsa, Tver Oblast (501 species, 10 km2); 06. Man-Pupu-Ner, Komi Republic (182 species, ca. 300 km2); 07. Middle Lyaga, Komi Republic (143 species, ca. 300 km2); 08. Utrish, Krasnodar Krai (933 species, 195 km2); 09. Yasnaya Polyana, Tula Oblast (236 species, 2.2 km2); 10. Bogolyubovsky Lug, Vladimir Oblast (289 species, 1.7 km2); 11. Tsaritsyno, City of Moscow (359 species, 5.3 km2); 12. Patakino, Vladimir Oblast (312 species, 1.1 km2). According to the GBIF taxonomic backbone, the dataset covers 1,806 species, 669 genera and 127 families of tracheophytes.

Flora of Vladimir Oblast, Russia: an updated grid dataset (1867-2020).

BACKGROUND: The dataset covers wild tracheophytes (native species, naturalised aliens and casuals) of Vladimir Oblast, Russia. It includes only one occurrence per species per grid square, thereby recently confirmed earlier records are not duplicated. Georeferences are based on the WGS84 grid scheme with 342 squares with areas ranging from 94.7 km2 in the northernmost part to 98.2 km2 on the southern boundary (5' lat. × 10' long.). Each occurrence is linked to the corresponding grid square centroid, therefore actual coordinates, habitat details and voucher information are unavailable. In late 2011, the earlier version of the dataset was used for the production of grid maps in the standard "Flora of Vladimir Oblast: checklist and atlas". Additional records, obtained during field excursions of 2012 and 2013, were fully included in the "Flora of Vladimir Oblast: grid data analysis". The stable version of the dataset with 123,054 grid records (as of 1867-2013) was published in GBIF in 2017. NEW INFORMATION: Data obtained in the field during 2014-2020, as well as those extracted from recently published sources, were digitised, structured and finally published in GBIF in April 2021. The last update added 7,000 new grid records. Currently, "Flora of Vladimir Oblast, Russia: an updated grid dataset (1867-2020)" contains 130,054 unique occurrences of 1,465 vascular plant taxa (species, hybrids, species aggregates) from Vladimir Oblast and tiny parts of the adjacent areas. The average number of grid records has grown over the seven years from 363 to 380 species. The grid occurrences are largely based on the field studies by the author, performed during 1999-2020 (121,737 records), as well as on data extracted from the relevant literature, unpublished sources, herbarium collections and citizen science projects (8,317 records). The taxonomic backbone of the occurrence grid dataset follows the accompanying checklist dataset to ensure correct cross-linking of the names. As of April 2021, the dataset on the Vladimir Oblast flora represents the fourth largest dataset on vascular plants of Russia published in GBIF.

Luminore CopperTouch Surface Coating Effectively Inactivates SARS-CoV-2, Ebola Virus, and Marburg Virus In Vitro.

We investigated the ability of Luminore CopperTouch copper and copper-nickel surfaces to inactivate filoviruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The copper and copper-nickel surfaces inactivated 99.9% of Ebola and Marburg viruses after 30 min, and the copper surfaces inactivated 99% of SARS-CoV-2 in 2 h. These data reveal that Ebola virus, Marburg virus, and SARS-CoV-2 are inactivated by exposure to copper ions, validating Luminore CopperTouch as an efficacious tool for infection control.

MHA Herbarium: Eastern European collections of vascular plants.

BACKGROUND: World herbaria with 387.5M specimens (Thiers 2019) are being rapidly digitised. At least 79.9M plant specimens (20.6%) are already databased throughout the globe in the standard form of GBIF-mediated data. The contribution of smaller herbaria has been steadily growing over the last few years due to cost reduction, usage of platforms and solutions developed by the leaders. A web-resource the Moscow Digital Herbarium (Seregin 2020b) was launched by the Lomonosov Moscow State University in October, 2016 for publication of specimens imaged and databased in the Moscow University Herbarium (MW). As of 31 December 2018, the web-portal included 968,031 images of 971,732 specimens digitised in MW. This dataset is available in GBIF (Seregin 2020). The global trend is largely the same in Russia, where a dozen herbaria started to scan their holdings after imaging of the nation's second largest herbarium (Kislov et al. 2017, Kovtonyuk et al. 2019, Seregin 2020a). In 2019, we started to use Moscow Digital Herbarium as a web-repository for digitised herbarium specimens from some Russian collections, starting with the Herbarium of Tsitsin Main Botanical Gaden, Russian Academy of Sciences (MHA). Due to this, a single-university system became a multi-institutional consortium in April 2019 (Seregin 2020a). The dataset of the Moscow collections and partly of the Eastern European collections of the MHA Herbarium is now available in GBIF (Seregin and Stepanova 2020). NEW INFORMATION: MHA Herbarium imaged 64,008 specimens from Moscow Region and partly from other regions of Eastern Europe at 600 dpi and provided key metadata. These data are now fully available in the Moscow Digital Herbarium and GBIF. Complete georeferencing of the specimens from the City of Moscow was a key task in 2020. As of May 2020, 50,324 specimens, including 49,732 specimens from Russia, have been georeferenced (78.6%) and 39,448 specimens have fully-captured label transcriptions (61.6%). Based on these data, we give a detailed overview of the collections including spatial, temporal and taxonomic description of the dataset.

"Flora of Russia" on iNaturalist: a dataset.

BACKGROUND: The "Flora of Russia" project on iNaturalist brought together professional scientists and amateur naturalists from all over the country. Over 10,000 people were involved in the data collection. NEW INFORMATION: Within 20 months, the participants accumulated 750,143 photo observations of 6,857 species of the Russian flora. This constitutes the largest dataset of open spatial data on the country's biodiversity and a leading source of data on the current state of the national flora. About 87% of all project data, i.e. 652,285 observations, are available under free licences (CC0, CC-BY, CC-BY-NC) and can be freely used in scientific, educational and environmental activities.

A single mutation (V64G) within the RING Domain of Z attenuates Junin virus.

Junin virus (JUNV) is a New World arenavirus that is the causative agent of Argentine hemorrhagic fever (AHF). Candid#1 (Can) is a live-attenuated vaccine strain of JUNV that since its introduction has resulted in a marked decrease in AHF incidence within the endemic regions of the Pampas in Argentina. Originally, the viral determinants and mechanisms of Can attenuation were not well understood. Recent work has identified the glycoprotein as the major attenuating factor for Can. The establishment of attenuating strategies based on any of the other viral proteins, however, has not been pursued. Here, we document the role of Can Z resulting in incompatibilities with wild type JUNV that results in decreased growth in vitro. In addition, this incompatibility results in attenuation of the virus in the guinea pig model. Further, we identify a single mutation (V64G) in the Z protein that is able to confer this demonstrated attenuation. By establishing and characterizing a novel attenuation strategy for New World mammarenaviruses, we hope to aid future vaccine development for related emerging pathogens including Machupo virus (MACV), Guanarito virus (GTOV), and Sabia virus (SABV).

Luminore CopperTouch™ surface coating effectively inactivates SARS-CoV-2, Ebola and Marburg viruses in vitro.

We investigated the ability of Luminore CopperTouch™ copper and copper-nickel surfaces to inactivate filoviruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). For this purpose, we compared viral titers in Vero cells from viral droplets exposed to copper surfaces for 30 min. The copper and copper-nickel surfaces inactivated 99.9% of the viral titer of both Ebola and Marburg viruses. The copper surfaces also inactivated 99% of SARS-CoV-2 titers in 2 hours to close to the limit of detection. These data add Ebolavirus, Marburgvirus, and SARS-CoV-2 (COVID-19) to the list of pathogens that can be inactivated by exposure to copper ions, validating Luminore CopperTouch™ technology (currently the only Environmental Protection Agency [EPA]-registered cold spray antimicrobial surface technology) as an efficacious, cost-friendly tool to improve infection control in hospitals, long-term care facilities, schools, hotels, buses, trains, airports, and other highly trafficked areas.

The Glycoprotein of the Live-Attenuated Junin Virus Vaccine Strain Induces Endoplasmic Reticulum Stress and Forms Aggregates prior to Degradation in the Lysosome.

Argentine hemorrhagic fever is a potentially lethal disease that is caused by Junin virus (JUNV). There are currently around 5 million individuals at risk of infection within regions of endemicity in Argentina. The live attenuated vaccine strain Candid #1 (Can) is approved for use in regions of endemicity and has substantially decreased the number of annual Argentine hemorrhagic fever (AHF) cases. The glycoprotein (GPC) gene is primarily responsible for attenuation of the Can strain, and we have shown that the absence of an N-linked glycosylation motif in the subunit G1 of the glycoprotein complex of Can, which is otherwise present in the wild-type pathogenic JUNV, causes GPC retention in the endoplasmic reticulum (ER). Here, we show that Can GPC aggregates in the ER of infected cells, forming incorrect cross-chain disulfide bonds, which results in impaired GPC processing into G1 and G2. The GPC fails to cleave into its G1 and G2 subunits and is targeted for degradation within lysosomes. Cells infected with the wild-type Romero (Rom) strain do not produce aggregates that are observed in Can infection, and the stress on the ER remains minimal. While the mutation of the N-linked glycosylation motif (T168A) is primarily responsible for the formation of aggregates, other mutations within G1 that occurred earlier in the passage history of the Can strain also contribute to aggregation of the GPC within the ER.IMPORTANCE The development of vaccines and therapeutics to combat viral hemorrhagic fevers remains a top priority within the Implementation Plan of the U.S. Department of Health and Human Services Public Health Emergency Medical Countermeasures Enterprise. The Can strain, derived from the pathogenic XJ strain of JUNV, has been demonstrated to be both safe and protective against AHF. While the vaccine strain is approved for use in regions of endemicity within Argentina, the mechanisms of Can attenuation have not been elucidated. A better understanding of the viral genetic determinants of attenuation will improve our understanding of the mechanisms contributing to disease pathogenesis and provide critical information for the rational design of live attenuated vaccine candidates for other viral hemorrhagic fevers.