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Objectives: Allied Health Professionals (AHPs) have a crucial role in reducing health inequalities. However, there is a lack of evidence regarding the ways they can fulfil this role. This rapid review explores the ways in which AHPs can decrease health care or health outcome inequalities; address inequalities in the social determinants of health; and support disadvantaged groups at an individual, organisational and system level. Study design: Rapid review following Cochrane criteria and narrative synthesis. Methods: MEDLINE, EMBASE, CINAHL, Web of Science and AMED were searched combined with grey literature, to identify quantitative or qualitative review articles published between January 2010 and February 2021. Results: From 8727 references, 36 met the inclusion criteria. The methodological quality of the studies was assessed with the AMSTAR tool and was generally low. Meta-analysis was not possible due to the heterogeneity of the studies, and a narrative synthesis was produced. Three themes emerged at patient and organisational level: 1) access to AHP services; 2) quality of care; and 3) social determinants of health. Two themes emerged at system level: 1) unequal workforce distribution and 2) lack of inclusive clinical guidelines. Conclusions: This rapid review offers a broad range of evidence on the ways AHPs can contribute to the reduction of inequalities in health care, both in terms of access and quality of care and in health outcomes. More research is needed to further understand the impact of AHPs on inequalities affecting specific groups and their contribution to equitable distribution of social determinants of health.

The silver-lipped pearl oyster, Pinctada maxima, is an important aquaculture species extensively farmed in tropical Australia and Southeast Asia to produce “South Sea” pearls. The pearling industry in Australia is in the process of implementing breeding programs targeting improved disease resistance, growth and pearl quality, and as a result genetic tools that can be readily implemented into breeding programs have been developed. However, to date there is no routine single nucleotide polymorphism (SNP) pedigree reconstruction tool publicly available.In this study, we isolated and assessed a panel of 934 genome-wide DArTcap™ SNPs for their utility and discriminatory power in resolving pedigree relationships in P. maxima. The panel was assessed for its power to assign parentage through in silico simulations of 1,000 progeny in a mass-spawning scenario, based on genotypes of 100 broodstock (1:1 sex ratio) and allowing for random missing genotypes in individuals (0–40%). Assignment success was then assessed across various pedigree panel sizes (25–934 SNPs ranked from the highest minor allele frequency). Simulations showed that a SNP panel comprising 50 or more SNPs had high pedigree resolution and the power to correctly assign progeny to a parent-progeny trio with 95 % confidence. In the simulation examining the effect of missing genotypes on assignment success, even where 40 % of alleles were missing in individuals only 150 SNPs were required in the panel to accurately assign parentage. Progeny from a commercial hatchery mass spawn involving 184 candidate parents were also assigned to their parents using the SNP panel and identified 79 full-sib families and genetic contributions from 33 sires and 11 dams. A highly skewed representation of family distributions in the cohort was observed which highlights the challenge of using a mass-spawning approach to the capture and identification of genetic diversity in P. maxima breeding programs.

Conventional methods for detecting differences in microsatellite repeat lengths rely on electrophoretic fractionation on long denaturing polyacrylamide gels, a time-consuming and labor-intensive method. Therefore, there is a need for the development of new and rapid approaches to routinely detect such length polymorphisms. The advent of techniques allowing the coupling of DNA molecules to solid surfaces has provided new prospects in the area of mutation detection. We describe here the development and optimization of the ligase-assisted spacer addition (LASA) method, a novel and rapid procedure based on an ELISA format to measure microsatellite repeat lengths. The LASA assay was successfully applied to a set of 11 bird samples to assess its capabilities as a genotyping method.

PCR-based cancer diagnosis requires detection of rare mutations in k-ras, p53 or other genes. The assumption has been that mutant and wild-type sequences amplify with near equal efficiency, so that they are eventually present in proportions representative of the starting material. Work on factor IX suggests that this assumption is invalid for one case of near-sequence identity. To test the generality of this phenomenon and its relevance to cancer diagnosis, primers distant from point mutations in p53 and k-ras were used to amplify wild-type and mutant sequences from these genes. A substantial bias against PCR amplification of mutants was observed for two regions of the p53 gene and one region of k-ras. For kras and p53, bias was observed when the wild-type and mutant sequences were amplified separately or when mixed in equal proportions before PCR. Bias was present with proofreading and non-proofreading polymerases. Mutant and wild-type segments of the factor V, cystic fibrosis transmembrane conductance regulator and prothrombin genes were amplified and did not exhibit PCR bias. Therefore, the assumption of equal PCR efficiency for point mutant and wild-type sequences is invalid in several systems. Quantitative or diagnostic PCR will require validation for each locus, and enrichment strategies may be needed to optimize detection of mutants.

A single bubble was generated and levitated in a high-intensity sound field within a spherical flask excited in its fundamental mode. Under optimum experimental conditions the bubble was observed to emit light in the form of short flashes. This phenomenon is known as single bubble sonoluminescence (SBSL). Using this process, the emitted light from the bubble was monitored when solutions containing fluorescein, quinine and sodium, potassium and copper salts were placed in the cell. The results obtained indicated that reproducible signals related directly to the concentration of the species present in solution could be achieved using single bubble sonoluminescence. The results for the molecular species were compared with those obtained by fluorescence spectroscopy and, in the case of quinine, parallel determinations of concentration in a test solution were performed with consistent results. SBSL signals were also observed to exhibit a linear correlation with the concentration of several trace metal salts introduced to the solution in the measurement cell. However, it was not possible to demonstrate that the SBSL signals were derived from stimulated atomic emission or fluorescence, and it was concluded that the effect may result from an indirect effect involving the bubble excitation mechanism. [ABSTRACT FROM AUTHOR]

Donald R. Barnard from the USDA-Agricultural Research Service Center for Medical, Agricultural, and Veterinary Entomology at Gainesville in Florida, USA, describes West Nile virus, an emerging mosquito-borne disease agent in North America where it threatens human and animal health. Historically, it has caused encephalitis and death in Africa, Europe, the Middle East, west and central Asia, and Oceania. Preventing future outbreaks of WN fever requires effective virus surveillance and mosquito control technology. [ABSTRACT FROM AUTHOR]

Abstract Background Declining gait performance is seen in aging individuals, due to neural and systemic factors. Plasma biomarkers provide an accessible way to assess evolving brain changes; non-specific neurodegeneration (NfL, GFAP) or evolving Alzheimer’s disease (Aβ 42/40 ratio, P-Tau181). Methods In a population-based cohort of older adults, we evaluate the hypothesis that plasma biomarkers of neurodegeneration and Alzheimer’s Disease pathology are associated with worse gait performance. A sample of 2641 Mayo Clinic Study of Aging participants with measurements of plasma biomarkers and gait parameters was analyzed in this cross-sectional study. Linear regression models using plasma biomarkers as predictors of gait parameters and adjusted for age, sex, BMI, Charlson Comorbidity Index, and cognitive diagnosis were evaluated. Results In this study multiple statistically significant relationships are observed for GFAP, NfL, and P-Tau181 with gait parameters. Each standard deviation increase in GFAP, NfL, and P-Tau181 is associated with a reduction in velocity of 2.100 (95% CI: −3.004, −1.196; p = 5.4 × 10−6), 4.400 (−5.292, -3.507; p = 9.5 × 10−22), and 2.617 (−3.414, −1.819; p = 1.5 × 10−10) cm/sec, respectively. Overall, NfL has the strongest associations with poor gait performance. Models with age interactions show that the strength of associations between the plasma biomarkers and the gait parameters became stronger with increasing age. There are no specific gait parameters that associate with individual plasma biomarkers. Conclusion Plasma biomarkers of neurodegeneration and Alzheimer’s Disease pathology are not only markers of cognitive decline but also indicate motor decline in the aging population.

Abstract Objective Prion disease classically presents with rapidly progressive dementia, leading to death within months of diagnosis. Advances in diagnostic testing have improved recognition of patients with atypical presentations and protracted disease courses, raising key questions surrounding the relationship between patterns of neurodegeneration and survival. We assessed the contribution of fluorodeoxyglucose (FDG‐PET) imaging for this purpose. Methods FDG‐PET were performed in 40 clinic patients with prion disease. FDG‐PET images were projected onto latent factors generated in an external dataset to yield patient‐specific eigenvalues. Eigenvalues were input into a clustering algorithm to generate data‐driven clusters, which were compared by survival time. Results Median age at FDG‐PET was 65.3 years (range 23–85). Median time from FDG‐PET to death was 3.7 months (range 0.3–19.0). Four data‐driven clusters were generated, termed “Neocortical” (n = 7), “Transitional” (n = 12), “Temporo‐parietal” (n = 13), and “Deep nuclei” (n = 6). Deep nuclei and transitional clusters had a shorter survival time than the neocortical cluster. Subsequent analyses suggested that this difference was driven by greater hypometabolism of deep nuclei relative to neocortical areas. FDG‐PET‐patterns were not associated with demographic (age and sex) or clinical (CSF total‐tau, 14‐3‐3) variables. Interpretation Greater hypometabolism within deep nuclei relative to neocortical areas associated with more rapid decline in patients with prion disease and vice versa. FDG‐PET informs large‐scale network physiology and may inform the relationship between spreading pathology and survival in patients with prion disease. Future studies should consider whether FDG‐PET may enrich multimodal prion disease prognostication models.

Abstract Two major barriers hinder the holistic understanding of subsurface critical zone (CZ) evolution and its impacts: (a) an inability to measure, define, and share information and (b) a societal structure that inhibits inclusivity and creativity. In contrast to the aboveground portion of the CZ, which is visible and measurable, the bottom boundary is difficult to access and quantify. In the context of these barriers, we aim to expand the spatial reach of the CZ by highlighting existing and effective tools for research as well as the “human reach” of CZ science by expanding who performs such science and who it benefits. We do so by exploring the diversity of vocabularies and techniques used in relevant disciplines, defining terminology, and prioritizing research questions that can be addressed. Specifically, we explore geochemical, geomorphological, geophysical, and ecological measurements and modeling tools to estimate CZ base and thickness. We also outline the importance of and approaches to developing a diverse CZ workforce that looks like and harnesses the creativity of the society it serves, addressing historical legacies of exclusion. Looking forward, we suggest that to grow CZ science, we must broaden the physical spaces studied and their relationships with inhabitants, measure the “deep” CZ and make data accessible, and address the bottlenecks of scaling and data‐model integration. What is needed—and what we have tried to outline—are common and fundamental structures that can be applied anywhere and used by the diversity of researchers involved in investigating and recording CZ processes from a myriad of perspectives.

CXOM31 J004252.030+413107.87 is one of the brightest X-ray sources within the D25 region of M31, and associated with a globular cluster known as B135; we therefore call this X-ray source XB135. XB135 is a low-mass X-ray binary (LMXB) that apparently exhibited hard state characteristics at 0.3–10 keV luminosities 4–6 erg s−1, and the hard state is only observed below ∼10% Eddington. If true, the accretor would be a high-mass black hole (BH) (≳50 ); such a BH may be formed from direct collapse of a metal-poor, high-mass star, and the very low metallicity of B135 (0.015 ) makes such a scenario plausible. We have obtained new XMM-Newton and Chandra HRC observations to shed light on the nature of this object. We find from the HRC observation that XB135 is a single point source located close to the center of B135. The new XMM-Newton spectrum is consistent with a rapidly spinning ∼10–20 BH in the steep power law or thermal dominant state, but inconsistent with the hard state that we previously assumed. We cannot formally reject three component emission models that have been associated with high luminosity neutron star (NS) LMXBs (known as Z-sources); however, we prefer a BH accretor. We note that deeper observation of XB135 could discriminate against an NS accretor. [ABSTRACT FROM AUTHOR]

Genetic mutations causative of frontotemporal lobar degeneration (FTLD) are highly predictive of a specific proteinopathy, but there exists substantial inter-individual variability in their patterns of network degeneration and clinical manifestations. We collected clinical and 18Fluorodeoxyglucose-positron emission tomography (FDG-PET) data from 39 patients with genetic FTLD, including 11 carrying the C9orf72 hexanucleotide expansion, 16 carrying a MAPT mutation and 12 carrying a GRN mutation. We performed a spectral covariance decomposition analysis between FDG-PET images to yield unbiased latent patterns reflective of whole brain patterns of metabolism (“eigenbrains” or EBs). We then conducted linear discriminant analyses (LDAs) to perform EB-based predictions of genetic mutation and predominant clinical phenotype (i.e., behavior/personality, language, asymptomatic). Five EBs were significant and explained 58.52 % of the covariance between FDG-PET images. EBs indicative of hypometabolism in left frontotemporal and temporo-parietal areas distinguished GRN mutation carriers from other genetic mutations and were associated with predominant language phenotypes. EBs indicative of hypometabolism in prefrontal and temporopolar areas with a right hemispheric predominance were mostly associated with predominant behavioral phenotypes and distinguished MAPT mutation carriers from other genetic mutations. The LDAs yielded accuracies of 79.5 % and 76.9 % in predicting genetic status and predominant clinical phenotype, respectively. A small number of EBs explained a high proportion of covariance in patterns of network degeneration across FTLD-related genetic mutations. These EBs contained biological information relevant to the variability in the pathophysiological and clinical aspects of genetic FTLD, and for offering valuable guidance in complex clinical decision-making, such as decisions related to genetic testing.

Abstract Understanding the severity and extent of near surface critical zone (CZ) disturbances and their ecosystem response is a pressing concern in the face of increasing human and natural disturbances. Predicting disturbance severity and recovery in a changing climate requires comprehensive understanding of ecosystem feedbacks among vegetation and the surrounding environment, including climate, hydrology, geomorphology, and biogeochemistry. Field surveys and satellite remote sensing have limited ability to effectively capture the spatial and temporal variability of disturbance and CZ properties. Technological advances in remote sensing using new sensors and new platforms have improved observations of changes in vegetation canopy structure and productivity; however, integrating measures of forest disturbance from various sensing platforms is complex. By connecting the potential for remote sensing technologies to observe different CZ disturbance vectors, we show that lower severity disturbance and slower vegetation recovery are more difficult to quantify. Case studies in montane forests from the western United States highlight new opportunities, including evaluating post‐disturbance forest recovery at multiple scales, shedding light on understory vegetation regrowth, detecting specific physiological responses, and refining ecohydrological modeling. Learning from regional CZ disturbance case studies, we propose future directions to synthesize fragmented findings with (a) new data analysis using new or existing sensors, (b) data fusion across multiple sensors and platforms, (c) increasing the value of ground‐based observations, (d) disturbance modeling, and (e) synthesis to improve understanding of disturbance.

Objectives: The aim of this study was to evaluate the value of digital subtraction angiography (DSA) in the diagnostic evaluation of a highly selected patient population presenting with pulse-synchronous tinnitus (PST). Methods: We retrospectively reviewed the charts of all patients referred for evaluation of possible vascular etiology of pulsatile tinnitus. Patients were evaluated with regards to presenting signs, comorbidities, non-invasive imaging results, angiographic findings and outcomes. Results: Fifteen patients underwent cerebral DSA. Dural arteriovenous fistula (dAVF) was identified in six patients, and five patients had other significant vascular pathology identified on DSA. Seven patients with ‘negative’ non-invasive imaging were found to have significant pathology on DSA. Conclusions: Catheter angiography may have a significant yield in appropriately selected patients presenting with pulse synchronous tinnitus.

Biogeochemical properties of soils play a crucial role in soil and stream chemistry throughout a watershed. How water interacts with soils during subsurface flow can have impacts on water quality, thus, it is fundamental to understand where and how certain soil water chemical processes occur within a catchment. In this study, ~200 soil samples were evaluated throughout a small catchment in the Front Range of Colorado, USA to examine spatial and vertical patterns in major soil solutes among different landscape units: riparian areas, alluvial/colluvial fans, and steep hillslopes. Solutes were extracted from the soil samples in the laboratory and analyzed for major cation (Li, K, Mg, Br, and Ca) and anion (F, Cl, NO2, NO3, PO4, and SO4) concentrations using ion chromatography. Concentrations of most solutes were greater in near surface soils (10 cm) than in deeper soils (100 cm) across all landscape units, except for F which increased with depth, suggestive of surface accumulation processes such as dust deposition or enrichment due to biotic cycling. Potassium had the highest variation between depths, ranging from 1.04 mg/l (100 cm) to 3.13 mg/l (10 cm) sampled from riparian landscape units. Nearly every solute was found to be enriched in riparian areas where vegetation was visibly denser, with higher mean concentrations than the hillslopes and fans, except for NO3 which had higher concentrations in the fans. Br, NO2, and PO4 concentrations were often below the detectable limit, and Li and Na were not variable between depths or landscape units. Ratioed stream water concentrations (K:Na, Ca:Mg, and NO3:Cl) vs. discharge relationships compared to the soil solute ratios indicated a hydraulic disconnection between the shallow soils (

Rhodopsin-related dominant retinitis pigmentosa is a degenerative disease of the retina of the eye for which there is no current treatment. In this study, the authors use a novel form of RNA-interference- artificial mirtrons- to slow retinal degeneration in a mouse model of the disease.

Recent advances in recombinant adeno-associated virus (rAAV) gene therapy for choroideremia show gene replacement to be a promising approach. It is, however, well known that contact of vector solution with plastic materials in the surgical device may result in non-specific adsorption with resulting loss of physical titer and/or level of protein expression and activity. Here we assessed the biocompatibility and stability of rAAV2-REP1 (Rab Escort Protein-1) before and following passage through the injection device over a period of time to mimic the clinical scenario. Three identical devices were screened using two concentrations of vector: high (1E+12 DNase-resistant particles [DRP]/mL) and low (1E+11 DRP/mL), to mimic high- and low-dose administrations of vector product. The low dose was prepared using either formulation buffer that contained 0.001% of a non-ionic surfactant (PF68) or balanced salt solution (BSS). We observed significant losses in the genomic titer of samples diluted with BSS for all time points. The addition of 0.001% PF68 did not, however, affect rAAV physical titer, or REP1 protein expression and biological activity. Hence we observed that neither the genomic titer nor the biological activity of a rAAV2-REP1-containing solution was affected following passage through the surgical device when PF68 was present as a surfactant and this was maintained over a period up to 10 h.

The 2015–2016 El Niño event ranks as one of the most severe on record in terms of the magnitude and extent of sea surface temperature (SST) anomalies generated in the tropical Pacific Ocean. Corresponding global impacts on the climate were expected to rival, or even surpass, those of the 1997–1998 severe El Niño event, which had SST anomalies that were similar in size. However, the 2015–2016 event failed to meet expectations for hydrologic change in many areas, including those expected to receive well above normal precipitation. To better understand how climate anomalies during an El Niño event impact soil moisture, we investigate changes in soil moisture in the humid tropics (between ±25∘) during the three most recent super El Niño events of 1982–1983, 1997–1998 and 2015–2016, using data from the Global Land Data Assimilation System (GLDAS). First, we use in situ soil moisture observations obtained from 16 sites across five continents to validate and bias-correct estimates from GLDAS (r2=0.54). Next, we apply a k-means cluster analysis to the soil moisture estimates during the El Niño mature phase, resulting in four groups of clustered data. The strongest and most consistent decreases in soil moisture occur in the Amazon basin and maritime southeastern Asia, while the most consistent increases occur over eastern Africa. In addition, we compare changes in soil moisture to both precipitation and evapotranspiration, which showed a lack of agreement in the direction of change between these variables and soil moisture most prominently in the southern Amazon basin, the Sahel and mainland southeastern Asia. Our results can be used to improve estimates of spatiotemporal differences in El Niño impacts on soil moisture in tropical hydrology and ecosystem models at multiple scales.

ObjectiveSigmoid sinus (SS) stenosis is a complication of translabyrinthine approach. Velocity changes in the SS measured by intra-operative doppler ultrasound may help in identifying patients at risk for sinus occlusion.PatientsSS velocity was measured using doppler ultrasound prior to opening dura and again prior to placement of the abdominal fat graft.InterventionData collected included: patient age, surgical side, sinus dominance, tumor volume, intra-operative doppler ultrasound measurements, post-operative venous sinus imaging, anticoagulation, and morbidities and mortalities.Main Outcome MeasureSS patency and velocity.ResultsEight patients were included in the analysis (22 to 69 years). Four had left-sided and four had right-sided craniotomies. Sigmoid sinuses were either right-side dominant or co-dominant. The mean velocity ± standard deviation (SD) prior to dura opening and abdominal fat packing was 23.2 ± 11.3 and 25.5 ± 13.9 cm/s, respectively, p = 0.575. Post-operative Magnetic Resonance Venography (MRV) imaging showed four sigmoid sinus occlusions; seven patients showed sigmoid sinus stenosis, and one internal jugular vein occlusion. One patient had post-operative Computed Tomography Venography (CTV) only. Of the four patients with MRV occlusions, CTVs were performed with three showing occlusion and all four-showing stenosis. One patient with internal jugular vein occlusion on MRV received warfarin anticoagulation. There was one cerebrospinal fluid leak requiring ear closure, one small cerebellar infarct, and one with facial nerve palsy (House-Brackman Grade 3).ConclusionSS velocity changes before and after tumor resection were not predictive of sinus occlusion. We hypothesize that sinus occlusion may be caused by related factors other than thrombosis, such as external compression of the sinus secondary to abdominal fat grafting.

Internal water storage within trees can be a critical reservoir that helps trees overcome both short- and long-duration environmental stresses. We monitored changes in internal tree water storage in a ponderosa pine on daily and seasonal scales using moisture probes, a dendrometer, and time-lapse electrical resistivity imaging (ERI). These data were used to investigate how patterns of in-tree water storage are affected by changes in sapflow rates, soil moisture, and meteorologic factors such as vapor pressure deficit. Measurements of xylem fluid electrical conductivity were constant in the early growing season while inverted sapwood electrical conductivity steadily increased, suggesting that increases in sapwood electrical conductivity did not result from an increase in xylem fluid electrical conductivity. Seasonal increases in stem electrical conductivity corresponded with seasonal increases in trunk diameter, suggesting that increased electrical conductivity may result from new growth. On the daily scale, changes in inverted sapwood electrical conductivity correspond to changes in sapwood moisture. Wavelet analyses indicated that lag times between inverted electrical conductivity and sapflow increased after storm events, suggesting that as soils wetted, reliance on internal water storage decreased, as did the time required to refill daily deficits in internal water storage. We found short time lags between sapflow and inverted electrical conductivity with dry conditions, when ponderosa pine are known to reduce stomatal conductance to avoid xylem cavitation. A decrease in diel amplitudes of inverted sapwood electrical conductivity during dry periods suggest that the ponderosa pine relied on internal water storage to supplement transpiration demands, but as drought conditions progressed, tree water storage contributions to transpiration decreased. Time-lapse ERI- and wavelet-analysis results highlight the important role internal tree water storage plays in supporting transpiration throughout a day and during periods of declining subsurface moisture.

Abstract Objectives Adults born prematurely have an increased risk of early heart failure. The impact of prematurity on left and right ventricular function has been well documented, but little is known about the impact on the systemic vasculature. The goals of this study were to measure aortic stiffness and the blood pressure response to physiological stressors; in particular, normoxic and hypoxic exercise. Methods Preterm participants (n = 10) were recruited from the Newborn Lung Project Cohort and matched with term‐born, age‐matched subjects (n = 12). Aortic pulse wave velocity was derived from the brachial arterial waveform and the heart rate and blood pressure responses to incremental exercise in normoxia (21% O2) or hypoxia (12% O2) were evaluated. Results Aortic pulse wave velocity was higher in the preterm groups. Additionally, heart rate, systolic blood pressure, and pulse pressure were higher throughout the normoxic exercise bout, consistent with higher conduit artery stiffness. Hypoxic exercise caused a decline in diastolic pressure in this group, but not in term‐born controls. Conclusions In this first report of the blood pressure response to exercise in adults born prematurely, we found exercise‐induced hypertension relative to a term‐born control group that is associated with increased large artery stiffness. These experiments performed in hypoxia reveal abnormalities in vascular function in adult survivors of prematurity that may further deteriorate as this population ages.

The precise control of eye size is essential for normal vision. TMEM98 is a highly conserved and widely expressed gene which appears to be involved in eye size regulation. Mutations in human TMEM98 are found in patients with nanophthalmos (very small eyes) and variants near the gene are associated in population studies with myopia and increased eye size. As complete loss of function mutations in mouse Tmem98 result in perinatal lethality, we produced mice deficient for Tmem98 in the retinal pigment epithelium (RPE), where Tmem98 is highly expressed. These mice have greatly enlarged eyes that are very fragile with very thin retinas, compressed choroid and thin sclera. To gain insight into the mechanism of action we used a proximity labelling approach to discover interacting proteins and identified MYRF as an interacting partner. Mutations of MYRF are also associated with nanophthalmos. The protein is an endoplasmic reticulum-tethered transcription factor which undergoes autoproteolytic cleavage to liberate the N-terminal part which then translocates to the nucleus where it acts as a transcription factor. We find that TMEM98 inhibits the self-cleavage of MYRF, in a novel regulatory mechanism. In RPE lacking TMEM98, MYRF is ectopically activated and abnormally localised to the nuclei. Our findings highlight the importance of the interplay between TMEM98 and MYRF in determining the size of the eye.

This review highlights the recent evolution of the imaging, medical management, surgical options and endovascular therapies for symptomatic intracranial atherosclerotic disease (ICAD). Recent imaging developments including optical coherence tomography and other modalities to assess the intracranial arteries for symptomatic ICAD are reviewed, not only to diagnose ICAD but to determine if ICAD plaques have any high-risk features for treatment. Potential future developments in the treatment of ICAD are discussed, including the development of trackable drug-coated balloons for the cerebral circulation to treat primary or restenotic arteries, new iterations of self-expanding intracranial stents with easier delivery systems, and the re-examination of indirect surgical bypass techniques for revascularisation. In addition to these important technological developments, however, is the evolving evidence regarding the best treatment window for these techniques and additional factors in medical management which can improve patient outcomes in this devastating pathology.

Choroideremia (CHM) is a rare, X-linked recessive retinal dystrophy caused by mutations in the CHM gene. CHM is ubiquitously expressed in human cells and encodes Rab escort protein 1 (REP1). REP1 plays a key role in intracellular trafficking through the prenylation of Rab GTPases, a reaction that can be reproduced in vitro. With recent advances in adeno-associated virus (AAV) gene therapy for CHM showing gene replacement to be a promising approach, an assay to assess the biological activity of the vectors is of the uttermost importance. Here we sought to compare the response of two Rab proteins, RAB27A and RAB6A, to the incorporation of a biotinylated lipid donor in a prenylation reaction in vitro. First, we found the expression of REP1 to be proportional to the amount of recombinant AAV (rAAV)2/2-REP1 used to transduce the cells. Second, prenylation of RAB6A appeared to be more sensitive to REP1 protein expression than prenylation of RAB27A. Moreover, the method was reproducible in other cell lines. These results support the further development of a prenylation reaction using a biotinylated lipid donor and RAB6A to assess the biological activity of AAV vectors for CHM gene therapy. Keywords: AAV gene therapy, choroideremia, prenylation, potency assay

Few studies have quantified the differences between celerity and velocity of hillslope water flow and explained the processes that control these differences. Here, we asses these differences by combining a 24-day hillslope sprinkling experiment with a spatially explicit hydrologic model analysis. We focused our work on Watershed 10 at the H. J. Andrews Experimental Forest in western Oregon. Celerities estimated from wetting front arrival times were generally much faster than average vertical velocities of δ2H. In the model analysis, this was consistent with an identifiable effective porosity (fraction of total porosity available for mass transfer) parameter, indicating that subsurface mixing was controlled by an immobile soil fraction, resulting in the attenuation of the δ2H input signal in lateral subsurface flow. In addition to the immobile soil fraction, exfiltrating deep groundwater that mixed with lateral subsurface flow captured at the experimental hillslope trench caused further reduction in the δ2H input signal. Finally, our results suggest that soil depth variability played a significant role in the celerity–velocity responses. Deeper upslope soils damped the δ2H input signal, while a shallow soil near the trench controlled the δ2H peak in lateral subsurface flow response. Simulated exit time and residence time distributions with our hillslope hydrologic model showed that water captured at the trench did not represent the entire modeled hillslope domain; the exit time distribution for lateral subsurface flow captured at the trench showed more early time weighting.

The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) has been included in the transgene cassette of adeno-associated virus (AAV) in several gene therapy clinical trials, including those for inherited retinal diseases. However, the extent to which WPRE increases transgene expression in the retina is still unclear. To address this question, AAV2 vectors containing a reporter gene with and without WPRE were initially compared in vitro and subsequently in vivo by subretinal delivery in mice. In both instances, the presence of WPRE led to significantly higher levels of transgene expression as measured by fundus fluorescence, western blot, and immunohistochemistry. The two vectors were further compared in human retinal explants derived from patients undergoing clinically indicated retinectomy, where again the presence of WPRE resulted in an enhancement of reporter gene expression. Finally, an analogous approach using a transgene currently employed in a clinical trial for choroideremia delivered similar results both in vitro and in vivo, confirming that the WPRE effect is transgene independent. Our data fully support the inclusion of WPRE in ongoing and future AAV retinal gene therapy trials, where it may allow a therapeutic effect to be achieved at an overall lower dose of vector. Keywords: AAV2, WPRE, gene therapy, mouse retina, human retina, choroideremia

Previous studies have used fluorescently labelled cells to demonstrate the incorporation of transplanted photoreceptor precursors into the mouse retina. Here, the authors show that fluorescent proteins are passed between the host and transplanted cells rather than migration of donor cells into the retina.

Abstract Mountains are vital to ecosystems and human society given their influence on global carbon and water cycles. Yet the extent to which topography regulates montane forest carbon uptake and storage remains poorly understood. To address this knowledge gap, we compared forest aboveground carbon loading to topographic metrics describing energy balance and water availability across three headwater catchments of the Boulder Creek Watershed, Colorado, USA. The catchments range from 1800 to 3500 m above mean sea level with 46–102 cm/yr mean annual precipitation and −1.2° to 12.3°C mean annual temperature. In all three catchments, we found mean forest carbon loading consistently increased from ridges (27 ± 19 Mg C ha) to valley bottoms (60 ± 28 Mg C ha). Low topographic positions held up to 185 ± 76 Mg C ha, more than twice the peak value of upper positions. Toe slopes fostered disproportionately high net carbon uptake relative to other topographic positions. Carbon storage was on average 20–40 Mg C ha greater on north to northeast aspects than on south to southwest aspects, a pattern most pronounced in the highest elevation, coldest and wettest catchment. Both the peak and mean aboveground carbon storage of the three catchments, crossing an 11°C range in temperature and doubling of local precipitation, defied the expectation of an optimal elevation‐gradient climatic zone for net primary production. These results have important implications for models of forest sensitivity to climate change, as well as to predicted estimates of continental carbon reservoirs.

In order to accelerate deployment of distributed recycling by providing low-cost feed stocks of granulated post-consumer waste plastic, this study analyzes an open source waste plastic granulator system. It is designed, built, and tested for its ability to convert post-consumer waste, 3D printed products and waste into polymer feedstock for recyclebots of fused particle/granule printers. The technical specifications of the device are quantified in terms of power consumption (380 to 404 W for PET and PLA, respectively) and particle size distribution. The open source device can be fabricated for less than $2000 USD in materials. The experimentally measured power use is only a minor contribution to the overall embodied energy of distributed recycling of waste plastic. The resultant plastic particle size distributions were found to be appropriate for use in both recyclebots and direct material extrusion 3D printers. Simple retrofits are shown to reduce sound levels during operation by 4dB-5dB for the vacuum. These results indicate that the open source waste plastic granulator is an appropriate technology for community, library, maker space, fab lab, or small business−based distributed recycling.

The recent emergence and rapid geographic expansion of Zika virus (ZIKV) poses a significant challenge for public health. Although historically causing only mild febrile illness, recent ZIKV outbreaks have been associated with more severe neurological complications, such as Guillain-Barré syndrome and fetal microcephaly. Here we demonstrate that two contemporary (2015) ZIKV isolates from Puerto Rico and Brazil may have increased replicative fitness in human astrocytoma cells. Over a single infectious cycle, the Brazilian isolate replicates to higher titers and induces more severe cytopathic effects in human astrocytoma cells than the historical African reference strain or an early Asian lineage isolate. In addition, both contemporary isolates induce significantly more double-stranded RNA in infected astrocytoma cells, despite similar numbers of infected cells across isolates. Moreover, when we quantified positive- and negative-strand viral RNA, we found that the Asian lineage isolates displayed substantially more negative-strand replicative intermediates than the African lineage isolate in human astrocytoma cells. However, over multiple rounds of infection, the contemporary ZIKV isolates appear to be impaired in cell spread, infecting a lower proportion of cells at a low MOI despite replicating to similar or higher titers. Taken together, our data suggests that contemporary ZIKV isolates may have evolved mechanisms that allow them to replicate with increased efficiency in certain cell types, thereby highlighting the importance of cell-intrinsic factors in studies of viral replicative fitness.

Case summary A 10-year-old neutered male domestic shorthair cat presented for evaluation of acute onset of paraplegia with loss of nociception and thoracolumbar spine hyperesthesia and no history of trauma. Activated partial thromboplastin time (aPTT) was markedly prolonged, and specific coagulation factor testing revealed a factor IX level of 4% of normal activity, confirming the presence of mild hemophilia B. Prior abnormal bleeding had occurred at the time of castration as a kitten, as well as with laceration to a toe. Advanced imaging, including computed tomography (CT) and magnetic resonance imaging (MRI) of the thoracolumbar spine, confirmed the presence of multifocal intradural and intramedullary spinal cord hemorrhage through demonstration of focal ring enhancement on CT and multifocal areas of signal void on gradient echo T2* images on MRI. Despite factor IX supplementation through the use of fresh frozen plasma transfusions and normalization of the aPTT time, the cat’s neurological status did not improve. Owing to repeated urinary tract infections, with increasing resistance to antibiotic therapy, the cat was ultimately euthanized. Post-mortem examination showed no evidence of another underlying primary pathology for the hematomyelia. Relevance and novel information To our knowledge, this case demonstrates the first reported occurrence of spontaneous hematomyelia secondary to hemophilia B in a cat.

Diadenosine tetraphosphate (Ap4A), a constituent of platelet dense granules, and its P1,P4-dithio and/or P2,P3-chloromethylene analogs, inhibit adenosine diphosphate (ADP)-induced platelet aggregation. We recently reported that these compounds antagonize both platelet ADP receptors, P2Y1 and P2Y12. The most active of those analogs, diadenosine 5',5″″-P1,P4-dithio-P2,P3-chloromethylenetetraphosphate, (compound 1), exists as a mixture of 4 stereoisomers.To separate the stereoisomers of compound 1 and determine their effects on platelet aggregation, platelet P2Y1 and P2Y12 receptor antagonism, and their metabolism in human plasma.We separated the 4 diastereomers of compound 1 by preparative reversed-phase chromatography, and studied their effect on ADP-induced platelet aggregation, P2Y1-mediated changes in cytosolic Ca2+, P2Y12-mediated changes in VASP phosphorylation, and metabolism in human plasma.The inhibition of ADP-induced human platelet aggregation and human platelet P2Y12 receptor, and stability in human plasma strongly depended on the stereo-configuration of the chiral P1- and P4-phosphorothioate groups, the SPSP diastereomer being the most potent inhibitor and completely resistant to degradation in plasma, and the RPRP diastereomer being the least potent inhibitor and with the lowest plasma stability. The inhibitory activity of SPRP diastereomers depended on the configuration of the pseudo-asymmetric carbon of the P2,P3-chloromethylene group, one of the configurations being significantly more active than the other. Their plasma stability did not differ significantly, being intermediate to that of the SPSP and the RPRP diastereomers.The presently-described stereoisomers have utility for structural, mechanistic, and drug development studies of dual antagonists of platelet P2Y1 and P2Y12 receptors.

Autosomal dominant optic atrophy (ADOA) is a slowly progressive optic neuropathy that has been associated with mutations of the OPA1 gene. In patients, the disease primarily affects the retinal ganglion cells (RGCs) and causes optic nerve atrophy and visual loss. A subset of RGCs are intrinsically photosensitive, express the photopigment melanopsin and drive non-image-forming (NIF) visual functions including light driven circadian and sleep behaviours and the pupil light reflex. Given the RGC pathology in ADOA, disruption of NIF functions might be predicted. Interestingly in ADOA patients the pupil light reflex was preserved, although NIF behavioural outputs were not examined. The B6; C3-Opa1(Q285STOP) mouse model of ADOA displays optic nerve abnormalities, RGC dendropathy and functional visual disruption. We performed a comprehensive assessment of light driven NIF functions in this mouse model using wheel running activity monitoring, videotracking and pupillometry. Opa1 mutant mice entrained their activity rhythm to the external light/dark cycle, suppressed their activity in response to acute light exposure at night, generated circadian phase shift responses to 480 nm and 525 nm pulses, demonstrated immobility-defined sleep induction following exposure to a brief light pulse at night and exhibited an intensity dependent pupil light reflex. There were no significant differences in any parameter tested relative to wildtype littermate controls. Furthermore, there was no significant difference in the number of melanopsin-expressing RGCs, cell morphology or melanopsin transcript levels between genotypes. Taken together, these findings suggest the preservation of NIF functions in Opa1 mutants. The results provide support to growing evidence that the melanopsin-expressing RGCs are protected in mitochondrial optic neuropathies.

Adeno-associated viral vectors (AAV) have been shown to be safe in the treatment of retinal degenerations in clinical trials. Thus, improving the efficiency of viral gene delivery has become increasingly important to increase the success of clinical trials. In this study, structural domains of different rAAV serotypes isolated from primate brain were combined to create novel hybrid recombinant AAV serotypes, rAAV2/rec2 and rAAV2/rec3. The efficacy of these novel serotypes were assessed in wild type mice and in two models of retinal degeneration (the Abca4(-/-) mouse which is a model for Stargardt disease and in the Pde6b(rd1/rd1) mouse) in vivo, in primate tissue ex-vivo, and in the human-derived SH-SY5Y cell line, using an identical AAV2 expression cassette. We show that these novel hybrid serotypes can transduce retinal tissue in mice and primates efficiently, although no more than AAV2/2 and rAAV2/5 serotypes. Transduction efficiency appeared lower in the Abca4(-/-) mouse compared to wild type with all vectors tested, suggesting an effect of specific retinal diseases on the efficiency of gene delivery. Shuffling of AAV capsid domains may have clinical applications for patients who develop T-cell immune responses following AAV gene therapy, as specific peptide antigen sequences could be substituted using this technique prior to vector re-treatments.

Platelets are involved in the thromboses that are central to myocardial infarctions and ischemic strokes. Such adverse cardiovascular events have day/night patterns with peaks in the morning (~9 AM), potentially related to endogenous circadian clock control of platelet activation. The objective was to test if the human endogenous circadian system influences (1) platelet function and (2) platelet response to standardized behavioral stressors. We also aimed to compare the magnitude of any effects on platelet function caused by the circadian system with that caused by varied standardized behavioral stressors, including mental arithmetic, passive postural tilt and mild cycling exercise.We studied 12 healthy adults (6 female) who lived in individual laboratory suites in dim light for 240 h, with all behaviors scheduled on a 20-h recurring cycle to permit assessment of endogenous circadian function independent from environmental and behavioral effects including the sleep/wake cycle. Circadian phase was assessed from core body temperature. There were highly significant endogenous circadian rhythms in platelet surface activated glycoprotein (GP) IIb-IIIa, GPIb and P-selectin (6-17% peak-trough amplitudes; p ≤ 0.01). These circadian peaks occurred at a circadian phase corresponding to 8-9 AM. Platelet count, ATP release, aggregability, and plasma epinephrine also had significant circadian rhythms but with later peaks (corresponding to 3-8 PM). The circadian effects on the platelet activation markers were always larger than that of any of the three behavioral stressors.These data demonstrate robust effects of the endogenous circadian system on platelet activation in humans--independent of the sleep/wake cycle, other behavioral influences and the environment. The 9 AM timing of the circadian peaks of the three platelet surface markers, including platelet surface activated GPIIb-IIIa, the final common pathway of platelet aggregation, suggests that endogenous circadian influences on platelet function could contribute to the morning peak in adverse cardiovascular events as seen in many epidemiological studies.

Progress in unravelling the cellular and molecular basis of mammalian circadian regulation over the past decade has provided us with new avenues through which we can explore central nervous system disease. Deteriorations in measurable circadian output parameters, such as sleep/wake deficits and dysregulation of circulating hormone levels, are common features of most central nervous system disorders. At the core of the mammalian circadian system is a complex of molecular oscillations within the hypothalamic suprachiasmatic nucleus. These oscillations are modifiable by afferent signals from the environment, and integrated signals are subsequently conveyed to remote central neural circuits where specific output rhythms are regulated. Mutations in circadian genes in mice can disturb both molecular oscillations and measurable output rhythms. Moreover, systematic analysis of these mutants indicates that they can express an array of abnormal behavioural phenotypes that are intermediate signatures of central nervous system disorders. Furthermore, the response of these mutants to psychoactive drugs suggests that clock genes can modify a number of the brain's critical neurotransmitter systems. This evidence has led to promising investigations into clock gene polymorphisms in psychiatric disease. Preliminary indications favour the systematic investigation of the contribution of circadian genes to central nervous system disease.