Association between benzodiazepine coprescription and mortality in people on opioid replacement therapy: a population-based cohort study.

OBJECTIVE: To investigate the association between opioid replacement therapy (ORT) and benzodiazepine (BZD) coprescription and all-cause mortality compared with the prescription of ORT alone. DESIGN: Population-based cohort study. SETTING: Scotland, UK. PARTICIPANTS: Participants were people prescribed ORT between January 2010 and end of December 2020 aged 18 years or above. MAIN OUTCOME MEASURES: All-cause mortality, drug-related deaths and non-drug related deaths. SECONDARY OUTCOME: ORT continuous treatment duration. ANALYSIS: Cox regression with time-varying covariates. RESULTS: During follow-up, 5776 of 46 899 participants died: 1398 while on coprescription and 4378 while on ORT only. The mortality per 100 person years was 3.11 during coprescription and 2.34 on ORT only. The adjusted HR for all-cause mortality was 1.17 (1.10 to 1.24). The adjusted HR for drug-related death was 1.14 (95% CI, 1.04 to 1.24) and the hazard for death not classified as drug-related was 1.19 (95% CI, 1.09 to 1.30). CONCLUSION: Coprescription of BZDs in ORT was associated with an increased risk of all-cause mortality, although with a small effect size than the international literature. Coprescribing was also associated with longer retention in treatment. Risk from BZD coprescription needs to be balanced against the risk from illicit BZDs and unplanned treatment discontinuation. A randomised controlled trial is urgently needed to provide a clear clinical direction. TRIAL REGISTRATION NUMBER: NCT04622995.

Baseline characteristics of people experiencing homelessness with a recent drug overdose in the PHOENIx pilot randomised controlled trial.

BACKGROUND: Drug-related deaths in Scotland are the highest in Europe. Half of all deaths in people experiencing homelessness are drug related, yet we know little about the unmet health needs of people experiencing homelessness with recent non-fatal overdose, limiting a tailored practice and policy response to a public health crisis. METHODS: People experiencing homelessness with at least one non-fatal street drug overdose in the previous 6 months were recruited from 20 venues in Glasgow, Scotland, and randomised into PHOENIx plus usual care, or usual care. PHOENIx is a collaborative assertive outreach intervention by independent prescriber NHS Pharmacists and third sector homelessness workers, offering repeated integrated, holistic physical, mental and addictions health and social care support including prescribing. We describe comprehensive baseline characteristics of randomised participants. RESULTS: One hundred and twenty-eight participants had a mean age of 42 years (SD 8.4); 71% male, homelessness for a median of 24 years (IQR 12-30). One hundred and eighteen (92%) lived in large, congregate city centre temporary accommodation. A quarter (25%) were not registered with a General Practitioner. Participants had overdosed a mean of 3.2 (SD 3.2) times in the preceding 6 months, using a median of 3 (IQR 2-4) non-prescription drugs concurrently: 112 (87.5%) street valium (benzodiazepine-type new psychoactive substances); 77 (60%) heroin; and 76 (59%) cocaine. Half (50%) were injecting, 50% into their groins. 90% were receiving care from Alcohol and Drug Recovery Services (ADRS), and in addition to using street drugs, 90% received opioid substitution therapy (OST), 10% diazepam for street valium use and one participant received heroin-assisted treatment. Participants had a mean of 2.2 (SD 1.3) mental health problems and 5.4 (SD 2.5) physical health problems; 50% received treatment for physical or mental health problems. Ninety-one per cent had at least one mental health problem; 66% had no specialist mental health support. Participants were frail (70%) or pre-frail (28%), with maximal levels of psychological distress, 44% received one or no daily meal, and 58% had previously attempted suicide. CONCLUSIONS: People at high risk of drug-related death continue to overdose repeatedly despite receiving OST. High levels of frailty, multimorbidity, unsuitable accommodation and unmet mental and physical health care needs require a reorientation of services informed by evidence of effectiveness and cost-effectiveness. Trial registration UK Clinical Trials Registry identifier: ISRCTN 10585019.

Pharmacist and Homeless Outreach Engagement and Non-medical Independent prescribing Rx (PHOENIx): a study protocol for a pilot randomised controlled trial.

INTRODUCTION: The number of people experiencing homelessness (PEH) is increasing worldwide. Systematic reviews show high levels of multimorbidity and mortality. Integrated health and social care outreach interventions may improve outcomes. No previous studies have targeted PEH with recent drug overdose despite high levels of drug-related deaths and few data describe their health/social care problems. Feasibility work suggests a collaborative health and social care intervention (Pharmacist and Homeless Outreach Engagement and Non-medical Independent prescribing Rx, PHOENIx) is potentially beneficial. We describe the methods of a pilot randomised controlled trial (RCT) with parallel process and economic evaluation of PEH with recent overdose. METHODS AND ANALYSIS: Detailed health and social care information will be collected before randomisation to care-as-usual plus visits from a pharmacist and a homeless outreach worker (PHOENIx) for 6-9 months or to care-as-usual. The outcomes are the rates of presentations to emergency department for overdose or other causes and whether to progress to a definitive RCT: recruitment of ≥100 participants within 4 months, ≥60% of patients remaining in the study at 6 and 9 months, ≥60% of patients receiving the intervention, and ≥80% of patients with data collected. The secondary outcomes include health-related quality of life, hospitalisations, treatment uptake and patient-reported measures. Semistructured interviews will explore the future implementation of PHOENIx, the reasons for overdose and protective factors. We will assess the feasibility of conducting a cost-effectiveness analysis. ETHICS AND DISSEMINATION: The study was approved by South East Scotland National Health Service Research Ethics Committee 01. Results will be made available to PEH, the study funders and other researchers. TRIAL REGISTRATION NUMBER: ISRCTN10585019.

Novel Anti-Neuroinflammatory Properties of a Thiosemicarbazone-Pyridylhydrazone Copper(II) Complex.

Neuroinflammation has a major role in several brain disorders including Alzheimer's disease (AD), yet at present there are no effective anti-neuroinflammatory therapeutics available. Copper(II) complexes of bis(thiosemicarbazones) (CuII(gtsm) and CuII(atsm)) have broad therapeutic actions in preclinical models of neurodegeneration, with CuII(atsm) demonstrating beneficial outcomes on neuroinflammatory markers in vitro and in vivo. These findings suggest that copper(II) complexes could be harnessed as a new approach to modulate immune function in neurodegenerative diseases. In this study, we examined the anti-neuroinflammatory action of several low-molecular-weight, charge-neutral and lipophilic copper(II) complexes. Our analysis revealed that one compound, a thiosemicarbazone-pyridylhydrazone copper(II) complex (CuL5), delivered copper into cells in vitro and increased the concentration of copper in the brain in vivo. In a primary murine microglia culture, CuL5 was shown to decrease secretion of pro-inflammatory cytokine macrophage chemoattractant protein 1 (MCP-1) and expression of tumor necrosis factor alpha (Tnf), increase expression of metallothionein (Mt1), and modulate expression of Alzheimer's disease-associated risk genes, Trem2 and Cd33. CuL5 also improved the phagocytic function of microglia in vitro. In 5xFAD model AD mice, treatment with CuL5 led to an improved performance in a spatial working memory test, while, interestingly, increased accumulation of amyloid plaques in treated mice. These findings demonstrate that CuL5 can induce anti-neuroinflammatory effects in vitro and provide selective benefit in vivo. The outcomes provide further support for the development of copper-based compounds to modulate neuroinflammation in brain diseases.

Where the wild things were: intrinsic and extrinsic extinction predictors in the world's most depleted mammal fauna.

Preventing extinctions requires understanding macroecological patterns of vulnerability or persistence. However, correlates of risk can be nonlinear, within-species risk varies geographically, and current-day threats cannot reveal drivers of past losses. We investigated factors that regulated survival or extinction in Caribbean mammals, which have experienced the globally highest level of human-caused postglacial mammalian extinctions, and included all extinct and extant Holocene island populations of non-volant species (219 survivals or extinctions across 118 islands). Extinction selectivity shows a statistically detectable and complex body mass effect, with survival probability decreasing for both mass extremes, indicating that intermediate-sized species have been more resilient. A strong interaction between mass and age of first human arrival provides quantitative evidence of larger mammals going extinct on the earliest islands colonized, revealing an extinction filter caused by past human activities. Survival probability increases on islands with lower mean elevation (mostly small cays acting as offshore refugia) and decreases with more frequent hurricanes, highlighting the risk of extreme weather events and rising sea levels to surviving species on low-lying cays. These findings demonstrate the interplay between intrinsic biology, regional ecology and specific local threats, providing insights for understanding drivers of biodiversity loss across island systems and fragmented habitats worldwide.

Identifying the possibilities and pitfalls of conducting IUCN Red List assessments from remotely sensed habitat information based on insights from poorly known Cuban mammals.

The International Union for Conservation of Nature's Red List of Threatened Species (RLS) is the key global tool for objective, repeatable assessment of species' extinction risk status, and plays an essential role in tracking biodiversity loss and guiding conservation action. Satellite remote sensing (SRS) data sets on global ecosystem distributions and functioning show exciting potential for informing range-based RLS assessment, but their incorporation has been restricted by low temporal resolution and coverage of data sets, lack of incorporation of degradation-driven habitat loss, and noninclusion of assumptions related to identification of changing habitat distributions for taxa with varying habitat dependency and ecologies. For poorly known mangrove-associated Cuban hutias (Mesocapromys spp.), we tested the impact of possible assumptions regarding these issues on range-based RLS assessment outcomes. Specifically, we used annual (1985-2018) Landsat data and land-cover classification and habitat degradation analyses across different internal time series slices to simulate range-based RLS assessments for our case study taxa to explore potential assessment uncertainty arising from temporal SRS data set coverage, incorporating proxies of (change in) habitat quality, and assumptions on spatial scaling of habitat extent for RLS parameter generation. We found extensive variation in simulated species-specific range-based RLS assessments, and this variation was mostly associated with the time series over which parameters were estimated. However, results of some species-specific assessments differed by up to 3 categories (near threatened to critically endangered) within the same time series, due to the effects of incorporating habitat quality and the spatial scaling used in RLS parameter estimation. Our results showed that a one-size-fits-all approach to incorporating SRS information in RLS assessment is inappropriate, and we urge caution in conducting range-based assessments with SRS for species for which habitat dependence on specific ecosystem types is incompletely understood. We propose novel revisions to parameter spatial scaling guidelines to improve integration of existing time series data on ecosystem change into the RLS assessment process.

La Lista Roja de Especies Amenazadas (LREA) de la Unión Internacional para la Conservación de la Naturaleza es la herramienta mundial más importante para la evaluación objetiva y repetible del estado de riesgo de extinción de una especie y juega un papel esencial en el seguimiento de la pérdida de la biodiversidad y en la orientación de las acciones de conservación. Los conjuntos de datos obtenidos por telemetría satelital (SRS) sobre la distribución y funcionamiento de los ecosistemas globales tienen un potencial emocionante para informar las evaluaciones de la LREA basadas en la extensión de la distribución de la especie, pero su incorporación dentro de los estudios ha estado restringida por la baja resolución temporal y la poca cobertura de los conjuntos de datos, la falta de inclusión de la pérdida de hábitat causada por la degradación y la nula inclusión de las suposiciones relacionadas con la identificación del cambio de hábitat de distribución para los taxones con una ecología y una dependencia por el hábitat variantes. Analizamos el impacto de las posibles suposiciones con respecto a los tres temas anteriores sobre los resultados de la evaluación de la LREA basada en la distribución de la jutía cubana (Mesocapromys spp.), una especie poco conocida y asociada con manglares. Específicamente, usamos los datos anuales (1985-2018) de Landsat y de la clasificación del uso de suelo y los análisis de degradación del hábitat en diferentes porciones de series temporales internas para simular las evaluaciones de la LREA basadas en la extensión para nuestro taxón de estudio y así explorar la incertidumbre potencial de la evaluación que surge de la cobertura del conjunto de datos SRS temporales. También incorporamos sustitutos de (cambio en) la calidad del hábitat y suposiciones sobre la escala espacial de la extensión del hábitat para la generación de parámetros de la LREA. Encontramos una variación extensa en las evaluaciones simuladas de la LREA específicas de especie y basadas en la extensión. Esta variación estuvo principalmente asociada con la serie temporal sobre la cual se estimaron los parámetros. Sin embargo, los resultados de algunas evaluaciones específicas de especie difirieron hasta en tres categorías (de casi amenazada hasta en peligro crítico) dentro de la misma serie temporal debido a los efectos de la incorporación de la calidad del hábitat y la escala espacial usadas en la estimación de parámetros de la LREA. Nuestros resultados muestran que un enfoque genérico para incorporar la información de SRS en la evaluación de la LREA es inapropiado e instamos precaución al realizar evaluaciones basadas en la extensión con datos SRS para especies cuya dependencia de hábitat por tipos específicos de ecosistemas no está entendida por completo. Proponemos que existan revisiones novedosas de las pautas para los parámetros de las escalas espaciales y así mejorar la integración de los datos existentes de series temporales sobre el cambio en el ecosistema dentro de los procesos de evaluación de RLS. Identificación de las Posibilidades y las Dificultades para la Realización de Evaluaciones de la Lista Roja de la UICN a partir de Información de Hábitat Detectada Remotamente con base en la Información sobre Mamíferos Cubanos Poco Conocidos.

Estimating changes and trends in ecosystem extent with dense time-series satellite remote sensing.

Quantifying trends in ecosystem extent is essential to understanding the status of ecosystems. Estimates of ecosystem loss are widely used to track progress toward conservation targets, monitor deforestation, and identify ecosystems undergoing rapid change. Satellite remote sensing has become an important source of information for estimating these variables. Despite regular acquisition of satellite data, many studies of change in ecosystem extent use only static snapshots, which ignores considerable amounts of data. This approach limits the ability to explicitly estimate trend uncertainty and significance. Assessing the accuracy of multiple snapshots also requires time-series reference data which is often very costly and sometimes impossible to obtain. We devised a method of estimating trends in ecosystem extent that uses all available Landsat satellite imagery. We used a dense time series of classified maps that explicitly accounted for covariates that affect extent estimates (e.g., time, cloud cover, and seasonality). We applied this approach to the Hukaung Valley Wildlife Sanctuary, Myanmar, where rapid deforestation is greatly affecting the lowland rainforest. We applied a generalized additive mixed model to estimate forest extent from more than 650 Landsat image classifications (1999-2018). Forest extent declined significantly at a rate of 0.274%/year (SE = 0.078). Forest extent declined from 91.70% (SE = 0.02) of the study area in 1999 to 86.52% (SE = 0.02) in 2018. Compared with the snapshot method, our approach improved estimated trends of ecosystem loss by allowing significance testing with confidence intervals and incorporation of nonlinear relationships. Our method can be used to identify significant trends over time, reduces the need for extensive reference data through time, and provides quantitative estimates of uncertainty.

Estimación de los Cambios y Tendencias en la Extensión de los Ecosistemas Mediante Teledetección Satelital de Series Temporales Densas Resumen Las tendencias de cuantificación de la extensión de los ecosistemas es esencial para el entendimiento de su estado. Las estimaciones de pérdidas de los ecosistemas se usan con amplitud para rastrear el progreso hacia los objetivos de conservación, monitorear la deforestación e identificar a los ecosistemas que están experimentando un cambio rápido. La teledetección satelital se ha transformado en una fuente importante de información para la estimación de estas variables. A pesar de la obtención de datos satelitales, muchos estudios sobre el cambio en la extensión de los ecosistemas usan solamente capturas estáticas, lo cual ignora cantidades considerables de datos. Esta estrategia limita la habilidad que se tiene para estimar explícitamente la incertidumbre e importancia de la tendencia. La valoración de la precisión de múltiples capturas también requiere datos de referencia de series temporales, lo cual es muy costoso e imposible de conseguir en algunos casos. Diseñamos un método para estimar las tendencias en la extensión de los ecosistemas que usa todas las imágenes satelitales disponibles en Landsat. Usamos una serie temporal densa de los mapas clasificados que considera explícitamente a las covarianzas que afectan a las estimaciones de la extensión (p.ej.: tiempo, cobertura de nubes y estacionalidad). Aplicamos esta estrategia en el Santuario de Vida Silvestre del Valle de Huakaung en Myanmar, en donde la deforestación acelerada está afectando enormemente a la selva de tierras bajas. Aplicamos también un modelo mixto, aditivo y generalizado para estimar la extensión del bosque a partir de más de 650 clasificaciones de imágenes en Landsat (1999 - 2018). La extensión del bosque declinó significativamente a una tasa de 0.274%/año (SE 0.078). La extensión del bosque declinó del 91.70% (SE 0.02) del área de estudio en 1999 a 86.52% (SE 0.02) en 2018. Si la comparamos con la estrategia de las capturas, nuestra estrategia mejoró las tendencias estimadas de la pérdida del ecosistema al permitir la evaluación de significancia con intervalos de confianza y la incorporación de relaciones no lineales. Nuestro método puede usarse para identificar las tendencias significativas a lo largo del tiempo; también reduce la necesidad de tener datos de referencia extensos a lo largo del tiempo y proporciona estimaciones cuantitativas de la incertidumbre.

Complementarity, completeness and quality of long-term faunal archives in an Asian biodiversity hotspot.

Long-term baselines on biodiversity change through time are crucial to inform conservation decision-making in biodiversity hotspots, but environmental archives remain unavailable for many regions. Extensive palaeontological, zooarchaeological and historical records and indigenous knowledge about past environmental conditions exist for China, a megadiverse country experiencing large-scale biodiversity loss, but their potential to understand past human-caused faunal turnover is not fully assessed. We investigate a series of complementary environmental archives to evaluate the quality of the Holocene-historical faunal record of Hainan Island, China's southernmost province, for establishing new baselines on postglacial mammalian diversity and extinction dynamics. Synthesis of multiple archives provides an integrated model of long-term biodiversity change, revealing that Hainan has experienced protracted and ongoing human-caused depletion of its mammal fauna from prehistory to the present, and that past baselines can inform practical conservation management. However, China's Holocene-historical archives exhibit substantial incompleteness and bias at regional and country-wide scales, with limited taxonomic representation especially for small-bodied species, and poor sampling of high-elevation landscapes facing current-day climate change risks. Establishing a clearer understanding of the quality of environmental archives in threatened ecoregions, and their ability to provide a meaningful understanding of the past, is needed to identify future conservation-relevant historical research priorities. This article is part of a discussion meeting issue 'The past is a foreign country: how much can the fossil record actually inform conservation?'

Changes in levels of cortical metabotropic glutamate 2 receptors with gender and suicide but not psychiatric diagnoses.

BACKGROUND: We previously reported that, compared to controls, there are lower levels of [3H]LY341495 binding to metabotropic 2/3 receptors (GRM2/3) in Brodmann's area (BA) 24, but not 17 or 46, from subjects with major depressive disorders (MDD) but not bipolar disorders (BD) or schizophrenia. To be able to better interpret these data we have now measured levels of GRM2 in two of these cortical regions. METHODS: Using a rabbit anti-metabotropic GRM2 monoclonal antibody with Western blotting we measured levels of GRM2 in BA 24 and 46 from subjects with MDD, BD, schizophrenia and controls (n = 15 per group). RESULTS: Compared to controls, levels of GRM2, normalised to ß-actin, did not differ in BA 24 or 46 from subjects with MDD, BD or schizophrenia (p from 0.36 to 0.79). Levels of GRM2 in BA 46, but not BA 24, were significantly higher in males compared to females (p < 0.01) and in suicide completers (p < 0.01) compare to death by other causes. LIMITATIONS: Our cohort sizes, whilst being comparable to many postmortem CNS studies, are relatively low. CONCLUSIONS: Our data suggests levels of GRM2 are not altered in two cortical regions from subjects with mood disorders or schizophrenia. Given we have found lower levels of [3H]LY341495 binding to GRM2/3 in BA 24 from subjects with MDD, our new data argues the lower levels of radioligand binding was due to lower levels of GRM3. Our data also suggests that glutamatergic activity through GRM2 in BA 46 may differ with gender and suicide ideation.

CuII(atsm) Attenuates Neuroinflammation.

Background: Neuroinflammation and biometal dyshomeostasis are key pathological features of several neurodegenerative diseases, including Alzheimer's disease (AD). Inflammation and biometals are linked at the molecular level through regulation of metal buffering proteins such as the metallothioneins. Even though the molecular connections between metals and inflammation have been demonstrated, little information exists on the effect of copper modulation on brain inflammation. Methods: We demonstrate the immunomodulatory potential of the copper bis(thiosemicarbazone) complex CuII(atsm) in an neuroinflammatory model in vivo and describe its anti-inflammatory effects on microglia and astrocytes in vitro. Results: By using a sophisticated in vivo magnetic resonance imaging (MRI) approach, we report the efficacy of CuII(atsm) in reducing acute cerebrovascular inflammation caused by peripheral administration of bacterial lipopolysaccharide (LPS). CuII(atsm) also induced anti-inflammatory outcomes in primary microglia [significant reductions in nitric oxide (NO), monocyte chemoattractant protein 1 (MCP-1), and tumor necrosis factor (TNF)] and astrocytes [significantly reduced NO, MCP-1, and interleukin 6 (IL-6)] in vitro. These anti-inflammatory actions were associated with increased cellular copper levels and increased the neuroprotective protein metallothionein-1 (MT1) in microglia and astrocytes. Conclusion: The beneficial effects of CuII(atsm) on the neuroimmune system suggest copper complexes are potential therapeutics for the treatment of neuroinflammatory conditions.

Optimal soil carbon sampling designs to achieve cost-effectiveness: a case study in blue carbon ecosystems.

Researchers are increasingly studying carbon (C) storage by natural ecosystems for climate mitigation, including coastal 'blue carbon' ecosystems. Unfortunately, little guidance on how to achieve robust, cost-effective estimates of blue C stocks to inform inventories exists. We use existing data (492 cores) to develop recommendations on the sampling effort required to achieve robust estimates of blue C. Using a broad-scale, spatially explicit dataset from Victoria, Australia, we applied multiple spatial methods to provide guidelines for reducing variability in estimates of soil C stocks over large areas. With a separate dataset collected across Australia, we evaluated how many samples are needed to capture variability within soil cores and the best methods for extrapolating C to 1 m soil depth. We found that 40 core samples are optimal for capturing C variance across 1000's of kilometres but higher density sampling is required across finer scales (100-200 km). Accounting for environmental variation can further decrease required sampling. The within core analyses showed that nine samples within a core capture the majority of the variability and log-linear equations can accurately extrapolate C. These recommendations can help develop standardized methods for sampling programmes to quantify soil C stocks at national scales.

The Role of Herbivory in Structuring Tropical Seagrass Ecosystem Service Delivery.

Seagrass meadows support key ecosystem services, via provision of food directly for herbivores, and indirectly to their predators. The importance of herbivores in seagrass meadows has been well-documented, but the links between food webs and ecosystem services in seagrass meadows have not previously been made explicit. Herbivores interact with ecosystem services - including carbon sequestration, cultural values, and coastal protection. Interactions can be positive or negative and depend on a range of factors including the herbivore identity and the grazing type and intensity. There can be unintended consequences from management actions based on a poor understanding of trade-offs that occur with complex seagrass-herbivore interactions. Tropical seagrass meadows support a diversity of grazers spanning the meso-, macro-, and megaherbivore scales. We present a conceptual model to describe how multiple ecosystem services are influenced by herbivore pressure in tropical seagrass meadows. Our model suggests that a balanced ecosystem, incorporating both seagrass and herbivore diversity, is likely to sustain the broadest range of ecosystem services. Our framework suggests the pathway to achieve desired ecosystem services outcomes requires knowledge on four key areas: (1) how size classes of herbivores interact to structure seagrass; (2) desired community and management values; (3) seagrass responses to top-down and bottom-up controls; (4) the pathway from intermediate to final ecosystem services and human benefits. We suggest research should be directed to these areas. Herbivory is a major structuring influence in tropical seagrass systems and needs to be considered for effective management of these critical habitats and their services.

Effect of Structural Modifications to Glyoxal-bis(thiosemicarbazonato)copper(II) Complexes on Cellular Copper Uptake, Copper-Mediated ATP7A Trafficking, and P-Glycoprotein Mediated Efflux.

Bis(thiosemicarbazonato)copper(II) complexes are of interest as potential therapeutics for cancer and neurodegenerative diseases as well as imaging agents for positron emission tomography (PET). The cellular uptake of six bis(thiosemcarbazonato)copper(II)complexes derived from glyoxal, with different functional groups Cu(gtsx) where x = different functional groups, was investigated in SKOV-3, HEK293, and HEK293 P-gp cell lines. Treatment of the cells with the copper complexes increased intracellular copper and increased levels of p-ERK due to activation of the Ras-Raf-MEK-ERK pathway. Treatment of SKOV-3 cells with low concentrations (µM) of two of the copper complexes led to trafficking of the endogenous copper transporter ATP7A from the Golgi network to the cell membrane. Experiments in HEK293 and HEK293-P-gp cells suggest that Cu(gtsm) and Cu(gtse) are substrates for the P-gp efflux protein but the complex with a pyrrolidine functional group, Cu(gtspyr), is not. A PET experiment in mice showed that [64Cu]Cu(gtspyr) has reasonable brain uptake but high liver uptake.

Adamantyl- and other polycyclic cage-based conjugates of desferrioxamine B (DFOB) for treating iron-mediated toxicity in cell models of Parkinson's disease.

The death of dopaminergic neurons is a major pathological hallmark of Parkinson's disease (PD). Elevated iron within the substantia nigra of the PD brain is thought to catalyze this neuronal death through hydroxyl radical-derived oxidative damage. Removing this excess iron presents a potential therapeutic strategy for PD. Seventeen derivatives of the non-toxic iron chelator desferrioxamine B (DFOB) were prepared by the conjugation of adamantyl- (1-4, 8-12), deconstructed adamantyl units (5-7), norborna(e)ne- (13-16) or bicyclo[2.2.2]octane-based (17) ancillary fragments to the terminal amine group. The range of experimental logP values of 1-17 (logP=0.15-2.82) was greater than water soluble DFOB (logP -2.29), with the increased hydrophobicity designed to improve cell membrane carriage to facilitate intracellular iron sequestration. The first activity screen showed compounds with methyl-substituted adamantyl (1-3), noradamantyl (5), or 1-pentylbicyclo[2.2.2]octane (17) ancillary groups significantly rescued iron-mediated oxidative stress in confluent PD-relevant SK-N-BE2-M17 neuroblastoma cells (M17 cells) exposed to 1,1'-dimethyl-4,4'-bipyridinium (paraquat, PQ) or H2O2. The second dose-dependence screen ranked 1-3 and 17 as the top candidates (EC50 ∼10µM) in the rescue of PQ-treated M17 cells. The ancillary fragments of 1-3 and 17 clustered in a region defined by a close-to-zero dipole moment, logP values of 2-2.8 and a surface area:volume ratio of 0.60-0.61. Results of iron leaching studies indicate that the compounds may be operating via mechanisms beyond solely removing intracellular iron. The DFOB conjugates with methyl-substituted adamantyl ancillary groups (1-3) were the top and most consistent performers in this class of compound designed for PD.

TDP-43 mutations causing amyotrophic lateral sclerosis are associated with altered expression of RNA-binding protein hnRNP K and affect the Nrf2 antioxidant pathway.

TAR DNA binding protein 43 (TDP-43) is a major disease-associated protein involved in the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). Our previous studies found a direct association between TDP-43 and heterogeneous nuclear ribonucleoprotein K (hnRNP K). In this study, utilizing ALS patient fibroblasts harboring a TDP-43M337V mutation and NSC-34 motor neuronal cell line expressing TDP-43Q331K mutation, we show that hnRNP K expression is impaired in urea soluble extracts from mutant TDP-43 cell models. This was confirmed in vivo using TDP-43Q331K and inducible TDP-43A315T murine ALS models. We further investigated the potential pathological effects of mutant TDP-43-mediated changes to hnRNP K metabolism by RNA binding immunoprecipitation analysis. hnRNP K protein was bound to antioxidant NFE2L2 transcripts encoding Nrf2 antioxidant transcription factor, with greater enrichment in TDP-43M337V patient fibroblasts compared to healthy controls. Subsequent gene expression profiling revealed an increase in downstream antioxidant transcript expression of Nrf2 signaling in the spinal cord of TDP-43Q331K mice compared to control counterparts, yet the corresponding protein expression was not up-regulated in transgenic mice. Despite the elevated expression of antioxidant transcripts, we observed impaired levels of glutathione (downstream Nrf2 antioxidant) in TDP-43M337V patient fibroblasts and astrocyte cultures from TDP-43Q331K mice, indicative of elevated oxidative stress and failure of some upregulated antioxidant genes to be translated into protein. Our findings indicate that further exploration of the interplay between hnRNP K (or other hnRNPs) and Nrf2-mediated antioxidant signaling is warranted and may be an important driver for motor neuron degeneration in ALS.

How many remnant gibbon populations are left on Hainan? Testing the use of local ecological knowledge to detect cryptic threatened primates.

For Critically Endangered "species of extreme rarity," there is an urgent need to clarify the potential survival of remnant populations. Such populations can be difficult to detect using standard field methods. Local ecological knowledge (LEK) represents an important alternative source of information, but anecdotal reports of rare or possibly extinct species can contain uncertainty and error. The Hainan gibbon (Nomascus hainanus), the world's rarest primate species, is confirmed to only survive as a tiny remnant population in Bawangling National Nature Reserve, China, but unverified gibbon sightings have been reported from other forest areas on Hainan. We conducted a large-scale community interview survey to gather new data on patterns of primate LEK from 709 respondents around seven reserves across Hainan, to investigate the possibility of gibbon survival outside Bawangling and assess whether LEK can provide useful information for conservation management of cryptic remnant populations. Comparative LEK data for gibbons and macaques are consistent with independent data on the relative status of these species across Hainan. Local awareness and experience of gibbons was low across Hainan, including at Bawangling, but we recorded recent anecdotal gibbon reports from most reserves. A follow-up field survey at Limushan Provincial Nature Reserve did not detect gibbons, however, and documented intensive wildlife exploitation within this reserve. All other surveyed landscapes showed some statistically lower levels of respondent awareness, experience, or sighting histories of gibbons compared to Bawangling, and are therefore considered biologically unlikely to support gibbons. Unverified LEK data can provide important insights into the possible status of cryptic remnant populations when assessed carefully and critically in relation to data from known populations.

Rehabilitating mangrove ecosystem services: A case study on the relative benefits of abandoned pond reversion from Panay Island, Philippines.

Mangroves provide vital climate change mitigation and adaptation (CCMA) ecosystem services (ES), yet have suffered extensive tropics-wide declines. To mitigate losses, rehabilitation is high on the conservation agenda. However, the relative functionality and ES delivery of rehabilitated mangroves in different intertidal locations is rarely assessed. In a case study from Panay Island, Philippines, using field- and satellite-derived methods, we assess carbon stocks and coastal protection potential of rehabilitated low-intertidal seafront and mid- to upper-intertidal abandoned (leased) fishpond areas, against reference natural mangroves. Due to large sizes and appropriate site conditions, targeted abandoned fishpond reversion to former mangrove was found to be favourable for enhancing CCMA in the coastal zone. In a municipality-specific case study, 96.7% of abandoned fishponds with high potential for effective greenbelt rehabilitation had favourable tenure status for reversion. These findings have implications for coastal zone management in Asia in the face of climate change.

Pyrrolidine dithiocarbamate activates the Nrf2 pathway in astrocytes.

BACKGROUND: Endogenous defense against oxidative stress is controlled by nuclear factor erythroid 2-related factor 2 (Nrf2). The normal compensatory mechanisms to combat oxidative stress appear to be insufficient to protect against the prolonged exposure to reactive oxygen species during disease. Counterbalancing the effects of oxidative stress by up-regulation of Nrf2 signaling has been shown to be effective in various disease models where oxidative stress is implicated, including Alzheimer's disease. Stimulation of Nrf2 signaling by small-molecule activators is an appealing strategy to up-regulate the endogenous defense mechanisms of cells. METHODS: Here, we investigate Nrf2 induction by the metal chelator and known nuclear factor-κB inhibitor pyrrolidine dithiocarbamate (PDTC) in cultured astrocytes and neurons, and mouse brain. Nrf2 induction is further examined in cultures co-treated with PDTC and kinase inhibitors or amyloid-beta, and in Nrf2-deficient cultures. RESULTS: We show that PDTC is a potent inducer of Nrf2 signaling specifically in astrocytes and demonstrate the critical role of Nrf2 in PDTC-mediated protection against oxidative stress. This induction appears to be regulated by both Keap1 and glycogen synthase kinase 3ß. Furthermore, the presence of amyloid-beta magnifies PDTC-mediated induction of endogenous protective mechanisms, therefore suggesting that PDTC may be an effective Nrf2 inducer in the context of Alzheimer's disease. Finally, we show that PDTC increases brain copper content and glial expression of heme oxygenase-1, and decreases lipid peroxidation in vivo, promoting a more antioxidative environment. CONCLUSIONS: PDTC activates Nrf2 and its antioxidative targets in astrocytes but not neurons. These effects may contribute to the neuroprotection observed for PDTC in models of Alzheimer's disease.