A single N6-methyladenosine site regulates lncRNA HOTAIR function in breast cancer cells.

N6-methyladenosine (m6A) modification of RNA regulates normal and cancer biology, but knowledge of its function on long noncoding RNAs (lncRNAs) remains limited. Here, we reveal that m6A regulates the breast cancer-associated human lncRNA HOTAIR. Mapping m6A in breast cancer cell lines, we identify multiple m6A sites on HOTAIR, with 1 single consistently methylated site (A783) that is critical for HOTAIR-driven proliferation and invasion of triple-negative breast cancer (TNBC) cells. Methylated A783 interacts with the m6A "reader" YTHDC1, promoting chromatin association of HOTAIR, proliferation and invasion of TNBC cells, and gene repression. A783U mutant HOTAIR induces a unique antitumor gene expression profile and displays loss-of-function and antimorph behaviors by impairing and, in some cases, causing opposite gene expression changes induced by wild-type (WT) HOTAIR. Our work demonstrates how modification of 1 base in an lncRNA can elicit a distinct gene regulation mechanism and drive cancer-associated phenotypes.

Funding and Implementation of Recovery Oriented Treatment Programs in the US from 2006-2020.

Background: Not much is known about funding for and implementation of Person-centered, long-term services - referred to as "recovery services." Methods: SAMSHA funding archives from 2004-2020 were analyzed using Latent Class Analysis (LCA). Results: All 50 states (plus DC and Guam) received about 482 recovery-based grants from 2004-2020 (total from 2004-2020 = $425 million vs. 63.3 ± 29.1 million in total SAMSHA funding per year on average). LCA showed 4 trends: peer focused (Pr(Class) = .09, 95%CI = 0.08, 0.10), treatment focused (Pr(Class) = 0.14, 95%CI = 0.12, 0.18), system focused (Pr(Class) = 0.57, 95% CI = 0.54, 0.59) and consumer focused (Pr(Class) = 0.19 (0.17, 0.21). Conclusions: Funding for recovery makes up a relatively low percentage of overall funding for substance prevention and treatment. Implications are discussed.

Rapid Characterization of Macroplastic Input and Leakage in the Ganges River Basin.

Efforts to understand macroplastic pollution have primarily focused on coastal and marine environments to the exclusion of freshwater, terrestrial, and urban ecosystems. To better understand macroplastics in the environment and their sources, a dual approach examining plastic input and leakage can be used. In this study, litter aggregation pathways at 40 survey sites with varying ambient population counts in the Ganges River Basin were surveyed in pre- and postmonsoon seasons. We examine active litter leakage using transect surveys of on-the-ground items, in conjunction with assessments of single-use plastic consumer products at the point of sale. We find that sites with low populations have a significantly higher number of littered items per 1,000 people than those with mid to high populations. Over 75% of litter items were plastics or multimaterial items containing plastic, and tobacco products and plastic food wrappers were the most recorded items. There was no significant variation of litter densities pre- and postmonsoon. Most single-use plastic consumer products were manufactured in-country, but approximately 40% of brands were owned by international companies. Stratified sampling of active litter input and consumer products provides a rapid, replicable snapshot of plastic use and leakage.

Cover crops differentially influenced nitrogen and phosphorus loss in tile drainage and surface runoff from agricultural fields in Ohio, USA.

Nitrogen (N) and phosphorus (P) loss from crop production agriculture is transported to adjacent and downstream water bodies, resulting in negative environmental impacts including harmful and nuisance algal blooms. Cover crops are a conservation management practice that replaces bare soil with vegetation outside of the cash crop growing season, purportedly reducing N and P loss by increasing water and nutrient demand in agroecosystems. In this study, we compared nitrate (NO3--N), total N (TN), dissolved reactive P (DRP), and total P (TP) loads in subsurface (tile) drainage and surface runoff from fields with cover crop management (CC) and fields without cover crop management (NoCC) using continuous monitoring data from 40 agricultural fields located throughout northcentral Ohio, United States (US). We found that average monthly tile NO3--N and TN loads from CC fields were ~50% less than NoCC fields, while average monthly tile discharge, DRP, and TP loads did not differ between CC and NoCC fields. Cover crops also did not significantly influence average monthly surface metrics. Cover crops reduced monthly totals of tile NO3--N and TN loads by ~1.0-2.6 kg N ha-1 from January to June (winter and spring), coinciding with critical periods of nutrient loss from agroecosystems in the midwestern US, but increased monthly totals of tile DRP (by 0.4-12.1 g DRP ha-1) and TP (by 1.2-31.6 g TP ha-1) loads during some months. We found similar patterns at the annual time scale whereby CC fields had lesser cumulative annual totals of tile NO3--N and TN but greater cumulative annual totals of tile DRP and TP. These results show that the influence of cover crops on N loads, but not P, were consistent across temporal scales of examination, demonstrating that cover crops effectively increased N demand and mitigated N losses from agricultural fields. The variable influence of cover crops on P loads underscores the need for greater understanding of the factors and mechanisms that control P loss in systems that include cover crop management. Furthermore, these findings stress the importance of identifying and selecting conservation management practices tailored to the natural resource concern.

Recruitment and allosteric stimulation of a histone-deubiquitinating enzyme during heterochromatin assembly.

Heterochromatin formation in budding yeast is regulated by the silent information regulator (SIR) complex. The SIR complex comprises the NAD-dependent deacetylase Sir2, the scaffolding protein Sir4, and the nucleosome-binding protein Sir3. Transcriptionally active regions present a challenge to SIR complex-mediated de novo heterochromatic silencing due to the presence of antagonistic histone post-translational modifications, including acetylation and methylation. Methylation of histone H3K4 and H3K79 is dependent on monoubiquitination of histone H2B (H2B-Ub). The SIR complex cannot erase H2B-Ub or histone methylation on its own. The deubiquitinase (DUB) Ubp10 is thought to promote heterochromatic silencing by maintaining low H2B-Ub at sub-telomeres. Here, we biochemically characterized the interactions between Ubp10 and the SIR complex machinery. We demonstrate that a direct interaction between Ubp10 and the Sir2/4 sub-complex facilitates Ubp10 recruitment to chromatin via a co-assembly mechanism. Using hydrolyzable H2B-Ub analogs, we show that Ubp10 activity is lower on nucleosomes compared with H2B-Ub in solution. We find that Sir2/4 stimulates Ubp10 DUB activity on nucleosomes, likely through a combination of targeting and allosteric regulation. This coupling mechanism between the silencing machinery and its DUB partner allows erasure of active PTMs and the de novo transition of a transcriptionally active DNA region to a silent chromatin state.

Microplastic ingestion ubiquitous in marine turtles.

Despite concerns regarding the environmental impacts of microplastics, knowledge of the incidence and levels of synthetic particles in large marine vertebrates is lacking. Here, we utilize an optimized enzymatic digestion methodology, previously developed for zooplankton, to explore whether synthetic particles could be isolated from marine turtle ingesta. We report the presence of synthetic particles in every turtle subjected to investigation (n = 102) which included individuals from all seven species of marine turtle, sampled from three ocean basins (Atlantic [ATL]: n = 30, four species; Mediterranean (MED): n = 56, two species; Pacific (PAC): n = 16, five species). Most particles (n = 811) were fibres (ATL: 77.1% MED: 85.3% PAC: 64.8%) with blue and black being the dominant colours. In lesser quantities were fragments (ATL: 22.9%: MED: 14.7% PAC: 20.2%) and microbeads (4.8%; PAC only; to our knowledge the first isolation of microbeads from marine megavertebrates). Fourier transform infrared spectroscopy (FT-IR) of a subsample of particles (n = 169) showed a range of synthetic materials such as elastomers (MED: 61.2%; PAC: 3.4%), thermoplastics (ATL: 36.8%: MED: 20.7% PAC: 27.7%) and synthetic regenerated cellulosic fibres (SRCF; ATL: 63.2%: MED: 5.8% PAC: 68.9%). Synthetic particles being isolated from species occupying different trophic levels suggest the possibility of multiple ingestion pathways. These include exposure from polluted seawater and sediments and/or additional trophic transfer from contaminated prey/forage items. We assess the likelihood that microplastic ingestion presents a significant conservation problem at current levels compared to other anthropogenic threats.

Molecular insights into RBR E3 ligase ubiquitin transfer mechanisms.

RING-in-between-RING (RBR) ubiquitin (Ub) ligases are a distinct class of E3s, defined by a RING1 domain that binds E2 Ub-conjugating enzyme and a RING2 domain that contains an active site cysteine similar to HECT-type E3s. Proposed to function as RING/HECT hybrids, details regarding the Ub transfer mechanism used by RBRs have yet to be defined. When paired with RING-type E3s, E2s perform the final step of Ub ligation to a substrate. In contrast, when paired with RBR E3s, E2s must transfer Ub onto the E3 to generate a E3~Ub intermediate. We show that RBRs utilize two strategies to ensure transfer of Ub from the E2 onto the E3 active site. First, RING1 domains of HHARI and RNF144 promote open E2~Ubs. Second, we identify a Ub-binding site on HHARI RING2 important for its recruitment to RING1-bound E2~Ub. Mutations that ablate Ub binding to HHARI RING2 also decrease RBR ligase activity, consistent with RING2 recruitment being a critical step for the RBR Ub transfer mechanism. Finally, we demonstrate that the mechanism defined here is utilized by a variety of RBRs.

Intrinsic disorder drives N-terminal ubiquitination by Ube2w.

Ubiquitination of the αN-terminus of protein substrates has been reported sporadically since the early 1980s. However, the identity of an enzyme responsible for this unique ubiquitin (Ub) modification has only recently been elucidated. We show the Ub-conjugating enzyme (E2) Ube2w uses a unique mechanism to facilitate the specific ubiquitination of the α-amino group of its substrates that involves recognition of backbone atoms of intrinsically disordered N termini. We present the NMR-based solution ensemble of full-length Ube2w that reveals a structural architecture unlike that of any other E2 in which its C terminus is partly disordered and flexible to accommodate variable substrate N termini. Flexibility of the substrate is critical for recognition by Ube2w, and either point mutations in or the removal of the flexible C terminus of Ube2w inhibits substrate binding and modification. Mechanistic insights reported here provide guiding principles for future efforts to define the N-terminal ubiquitome in cells.

Microplastics in beach sediments of the Azores archipelago, NE Atlantic.

Oceanic islands are exposed to plastic debris that has accumulated in the open ocean, particularly in the subtropical gyres. This study investigates the abundance and typology of microplastics (from 0.1 to 5 mm) on 19 sandy beaches spread across 8 oceanic islands of the Azores archipelago. Between January and April 2016, a total of 341 particles retrieved from all beaches, were identified as microplastics. The highest concentration (50.19 ± 21.93 particles kg-1 dw) was found in Terceira Island. Beach morphology and grain size were important factors explaining microplastic concentration. Fibres were the most dominant morphology recovered (80.9 %), followed by fragments (12.3 %). Fourier transform infrared spectroscopy (FTIR) revealed that 41 % of the fibres consisted of polyester and 60 % of the fragments were polyethylene. This research underlines the widespread contamination of microplastics in oceanic islands of the Atlantic Ocean.

Cory's shearwater as a key bioindicator for monitoring floating plastics.

The potential of using organisms as bioindicators of marine litter has been an area of general interest in multiple scientific and monitoring programs across the globe. Procellariiformes seabirds are particularly vulnerable to plastic contamination, which makes them a research focus group. This study investigated plastic ingestion in deceased fledglings and adults Cory's shearwaters (Calonectris borealis) collected over eight years (2015 to 2022) at two Atlantic archipelagos: the Azores and the Canaries. Necropsies were carried out in a total of 1,238 individuals showing a high prevalence of plastic ingestion (90%), with approximately 80% of items recovered from the gizzard. Fledglings carried greater plastic loads compared to adults, yet plastic morphologies were similar between both age classes. The temporal analyses conducted with generalised additive mixed-effect models revealed a distinct temporal trend in plastic numbers, but not in terms of plastic mass. In addition, the spatial analyses showed that Cory's shearwaters from the Canary Islands ingest a higher quantity of plastic and a greater proportion of threadlike items than the Azorean birds. These results suggest higher contamination at the NW Africa foraging grounds next to the Canaries and highlight fisheries as a potential source of marine litter in that region. On the other hand, the information gathered from the Azorean birds suggests they would be able to monitor changes in the composition of the plastic items floating in the North Atlantic Subtropical Gyre. Overall, our outcomes support the use of Cory's shearwater fledglings that are victims of light pollution as a key bioindicator of plastic contamination in the North Atlantic. For its policy application, the presented threshold value in combination with the assessment method will enable effective tracking of floating plastic litter in the framework of the MSFD and OSPAR.

Marine turtles as bio-indicators of plastic pollution in the eastern Mediterranean.

The loggerhead turtle (Caretta caretta) has been suggested as a bio-indicator species for plastic pollution. However, detailed investigations in the eastern Mediterranean are limited. Here, we present data from loggerhead turtles (2012-2022; n = 131) of which 42.7 % (n = 57) had ingested macroplastic (pieces ≥ 5 mm). Frequency of occurrence (%) was not found to have changed over time, with body size (CCL cm), between stranded or bycaught turtles, or with levels of digesta present. The characteristics of ingested plastic (n = 492) were largely sheetlike (62 %), clear (41 %) or white (25 %) and the most common polymers identified were Polypropylene (37 %) and Polyethylene (35 %). Strong selectivity was displayed towards certain types, colours and shapes. Data are also presented for posthatchling turtles (n = 4), an understudied life stage. Much larger sample sizes will be needed for this species to be an effective bio-indicator, with the consideration of monitoring green turtles (Chelonia mydas) for the eastern Mediterranean recommended allowing a more holistic picture to be gathered.

Cetaceans playing with single-use plastics (SUPs): A widespread interaction with likely severe impacts.

Play is a common behaviour in wild cetaceans that includes the manipulation of natural, as well as artificial objects such as marine debris. Yet, very little is known about these interactions despite the potential impacts on cetacean health. We combined a detailed review of the scientific literature and social media with 12 years of observations to examine cetacean interactions with plastic litter. A total of 11 odontocete species (Tursiops truncatus, Stenella longirostris, Delphinus delphis, Grampus griseus, Steno bredanensis, Stenella frontalis, Sotalia guianensis, Pseudorca crassidens, Orcinus orca, Globicephala melas and Physeter macrocephalus) were documented in 59 events carrying or throwing plastic litter with their head and/or flippers suggesting a form of play. Interactions occurred in the Atlantic, Pacific, Indian Ocean, Mediterranean, and Red Sea, with single-use plastics composing the main typology registered. While these interactions appeared harmless to the observers, they can pose a significant risk through subsequent entanglement or ingestion.

Plastic pollution on remote islands: A baseline study of Príncipe, Gulf of Guinea.

Plastic is the most abundant marine anthropogenic debris in the ocean and is of serious global environmental concern. Projects aimed at monitoring plastic pollution quantities are key in evaluating the success of reduction. This study provides baseline information on the quantities of marine debris found on the island of Príncipe in the Gulf of Guinea in the Atlantic coast of Central Africa. A total of 13,196 items were collected with 64.5% of these items being plastic. Plastic pieces the size of 2.5-50 cm made up 20.8% of all plastic items, followed by plastic bags (13.3%) and fishing material (11.39%). The density of mesoplastic items (<25 and >5mm) ranged between 0 and 6.78 items m-2. Beach location, community presence, and beach level (for turtle-nesting beaches) were important factors in explaining differences in plastic abundance and density around the island. Our results highlight the significance of high plastic pollution accumulation on remote islands.

Connecting soil characteristics to edge-of-field water quality in Ohio.

Soil health and water quality improvement are major goals of sustainable agricultural management systems, yet the connections between soil health and water quality impacts remain unclear. In this study we conducted an initial exploratory assessment of the relationships between soil chemical, physical, and biological properties and edge-of-field water quality across a network of 40 fields in Ohio, USA. Discharge, dissolved reactive P (DRP), total P (TP), and nitrate (NO3 ) losses associated with precipitation events via surface runoff and tile drainage were monitored. Agronomic soil tests and a suite of soil health indicators were measured, then predictive relationships between the field average soil properties and tile drainage and surface runoff discharge and DRP, TP, and nitrate loads were explored with random forest and multiple linear regression approaches. Among the soil health indicators, water extractable C and N were consistently found to be positively related to tile nitrate loads, but other soil health indicators had little or inconsistent importance for water quality impacts. Several other soil properties were important predictors, particularly soil P pools for surface and tile DRP and TP losses as well as Mehlich-3 (M3) extractable Fe and Al for surface and tile discharge. Thus, we did not observe strong evidence that soil health was associated with improved edge-of-field water quality across the edge-of-field monitoring network. However, additional studies are needed to definitively test the relationships between a broader array of soil health metrics and water quality outcomes.

The distribution and characterisation of microplastics in air, surface water and sediment within a major river system.

Rivers are key pathways for the transfer of microplastics (MP) to marine environments. However, there are considerable uncertainties about the amount of microplastics transported by rivers to the ocean; this results in inaccuracies in our understanding of microplastic quantity and transport by freshwater systems. Additionally, it has been suggested that rivers may represent long-term sinks, with microplastics accumulating in sediment due to their high density or other biological, chemical, and physical factors. The atmosphere is also an important pathway by which airborne microplastics may enter aquatic habitats. Here, we compare for first time microplastics type and concentration in these key environmental mediums (air, water and sediment) along a major river (Ganges), from sea to source to understand 1) the abundance, 2) the spatial distribution, and 3) characteristics. Mean microplastic abundance settling from the atmosphere was 41.12 MP m2 day-1; while concentrations in sediment were 57.00 MP kg-1 and in water were 0.05 MP L-1. Across all sites and environmental mediums, rayon (synthetically altered cellulose) was the dominant polymer (54-82 %), followed by acrylic (6-23 %) and polyester (9-17 %). Fibres were the dominant shape (95-99 %) and blue was the most common colour (48-79 %). Across water and sediment environmental mediums, the number of microplastics per sample increased from the source of the Ganges to the sea. Additionally, higher population densities correlated with increased microplastic abundance for air and water samples. We suggest that clothing is likely to be the prominent source of microplastics to the river system, influenced by atmospheric deposition, wastewater and direct input (e.g. handwashing of clothes in the Ganges), especially in high density population areas. However, we suggest that subsequent microplastic release to the marine environment is strongly influenced by polymer type and shape, with a large proportion of denser microplastics settling in sediment prior to the river discharging to the ocean.

Predicting the distribution and abundance of abandoned, lost or discarded fishing gear (ALDFG) in the deep sea of the Azores (North Atlantic).

Abandoned, lost, or discarded fishing gear (ALDFG), represents a significant percentage of the global plastic pollution, currently considered one of the major sources from sea-based activities. However, there is still limited understanding of the quantities of ALDFG present on the seafloor and their impacts. In this study, data on the presence of ALDFG was obtained from a large archive of seafloor video footage (351 dives) collected by different imaging platforms in the Azores region over 15 years (2006-2020). Most ALDFG items observed in the images relate to the local bottom longline fishery operating in the region, and include longlines but also anchors, weights, cables and buoys. A generalized additive mixed model (GAMM) was used to predict the distribution and abundance of ALDFG over the seafloor within the limits of the Azores Exclusive Economic Zone (EEZ) using a suite of environmental and anthropogenic variables. We estimated an average of 113 ± 310 items km-2 (597 ± 756 per km-2 above 1000 m depth), which could imply that over 20 million ALDFG items are present on the deep seafloor of the Azores EEZ. The resulting model identified potential hotspots of ALDFG along the seabed, some of them located over sensitive benthic habitats, such as specific seamounts. In addition, the interactions between ALDFG and benthic organisms were also analysed. Numerous entanglements were observed with several species of large anthozoans and sponges. The use of predictive distribution modelling for ALDFG should be regarded as a useful tool to support ecosystem-based management, which can provide indirect information about fishing pressure and allow the identification of potential high-risk areas. Additional knowledge about the sources, amounts, fates and impacts of ALDFG will be key to address the global issue of plastic pollution and the effects of fishing on marine ecosystems.

Interactions between marine megafauna and plastic pollution in Southeast Asia.

Southeast (SE) Asia is a highly biodiverse region, yet it is also estimated to cumulatively contribute a third of the total global marine plastic pollution. This threat is known to have adverse impacts on marine megafauna, however, understanding of its impacts has recently been highlighted as a priority for research in the region. To address this knowledge gap, a structured literature review was conducted for species of cartilaginous fishes, marine mammals, marine reptiles, and seabirds present in SE Asia, collating cases on a global scale to allow for comparison, coupled with a regional expert elicitation to gather additional published and grey literature cases which would have been omitted during the structured literature review. Of the 380 marine megafauna species present in SE Asia, but also studied elsewhere, we found that 9.1 % and 4.5 % of all publications documenting plastic entanglement (n = 55) and ingestion (n = 291) were conducted in SE Asian countries. At the species level, published cases of entanglement from SE Asian countries were available for 10 % or less of species within each taxonomic group. Additionally, published ingestion cases were available primarily for marine mammals and were lacking entirely for seabirds in the region. The regional expert elicitation led to entanglement and ingestion cases from SE Asian countries being documented in 10 and 15 additional species respectively, highlighting the utility of a broader approach to data synthesis. While the scale of the plastic pollution in SE Asia is of particular concern for marine ecosystems, knowledge of its interactions and impacts on marine megafauna lags behind other areas of the world, even after the inclusion of a regional expert elicitation. Additional funding to help collate baseline data are critically needed to inform policy and solutions towards limiting the interactions of marine megafauna and plastic pollution in SE Asia.

Ubiquitylation by Rab40b/Cul5 regulates Rap2 localization and activity during cell migration.

Cell migration is a complex process that involves coordinated changes in membrane transport and actin cytoskeleton dynamics. Ras-like small monomeric GTPases, such as Rap2, play a key role in regulating actin cytoskeleton dynamics and cell adhesions. However, how Rap2 function, localization, and activation are regulated during cell migration is not fully understood. We previously identified the small GTPase Rab40b as a regulator of breast cancer cell migration. Rab40b contains a suppressor of cytokine signaling (SOCS) box, which facilitates binding to Cullin5, a known E3 ubiquitin ligase component responsible for protein ubiquitylation. In this study, we show that the Rab40b/Cullin5 complex ubiquitylates Rap2. Importantly, we demonstrate that ubiquitylation regulates Rap2 activation as well as recycling of Rap2 from the endolysosomal compartment to the lamellipodia of migrating breast cancer cells. Based on these data, we propose that Rab40b/Cullin5 ubiquitylates and regulates Rap2-dependent actin dynamics at the leading edge, a process that is required for breast cancer cell migration and invasion.

Correction: Message in a bottle: Open source technology to track the movement of plastic pollution.

[This corrects the article DOI: 10.1371/journal.pone.0242459.].

Monitoring Different Social Media Platforms to Report Unplanned School Closures Due to Wildfires in California, October and December 2017.

OBJECTIVE: Researchers at the Centers for Disease Control and Prevention monitor unplanned school closure (USC) reports through online systematic searches (OSS) to assist public health emergency responses. We counted the additional reports identified through social media along with OSS to improve USC monitoring. METHODS: Facebook and Twitter data of public-school districts and private schools in counties affected by California wildfires in October and December of 2017 and January of 2018 were retrieved. We computed descriptive statistics and performed multivariable logistic regression for both OSS and social media data. RESULTS: Among the 362 public-school districts in wildfire-affected counties, USCs were identified for 115 (32%) districts, of which OSS identified 104 (90%), Facebook, 59 (52%), and Twitter, 37 (32%). These data correspond to 4622 public schools, among which USCs were identified for 888 (19.2%) schools, of which OSS identified 722 (81.3%), Facebook, 496 (55.9%), and Twitter, 312 (35.1%). Among 1289 private schools, USCs were identified for 104 schools, of which OSS identified 47 (45.2%), Facebook, 67 (64.4%), and Twitter, 29 (27.9%). USC announcements identified via social media, in addition to those via OSS, were 11 public school districts, 166 public schools, and 57 private schools. CONCLUSION: Social media complements OSS as additional resources for USC monitoring during disasters.