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Abstract Analysis of virus-like particles (VLPs) is an essential task in optimizing their implementation as vaccine antigens for virus-initiated diseases. Interrogating VLP collections for elasticity by probing with a rigid atomic force microscopy (AFM) tip is a potential method for determining VLP morphological changes. During VLP morphological change, it is not expected that all VLPs would be in the same state. This leads to the open question of whether VLPs may change in a continuous or stepwise fashion. For continuous change, the statistical distribution of observed VLP properties would be expected as a single distribution, while stepwise change would lead to a multimodal distribution of properties. This study presents the application of a Gaussian mixture model (GMM), fit by the Expectation-Maximization (EM) algorithm, to identify different states of VLP morphological change observed by AFM imaging.

Reviewing bodies have shown that a high percentage of SHS students tend to underperform in mathematics examinations. In that respect, we examined the influence of two teaching approaches on Ghanaian public Senior High School students' mathematical mindsets. A quantitative research design was used, that is, an ex post facto research design was used for this study. A total sample of 366 respondents was involved in this study. Two sets of questionnaires were employed for the study which are the students' perceptions of teaching approaches and mathematical mindset questionnaires. Descriptive analysis such as means and frequencies were used to analyze the data. The results revealed a possible influence of the teaching approaches students experienced and their mathematical mindset. That is, students taught with a student-centered approach were found to generally possess a growth mindset while those identified as possessing a more fixed mindset were mostly students taught using a more teacher-centered approach. The study recommended that teachers should adopt a student-centered approach to teaching since it has the potential to shape students' mindset toward a strong growth mindset, as students who have a growth mindset are more likely to succeed in school and beyond.

Lipid A is a fundamental Gram-negative outer membrane component and the essential element of lipopolysaccharide (endotoxin), a potent immunostimulatory molecule. This work describes the metabolic adaptation of the lipid A acyl structure by Psychrobacter cryohalolentis at various temperatures in its facultative psychrophilic growth range, as characterized by MALDI-TOF MS and FAME GC-MS. It also presents the first elucidation of lipid A structure from the Colwellia genus, describing lipid A from strains of Colwellia hornerae and Colwellia piezophila, which were isolated as primary cultures from Arctic fast sea ice and identified by 16S rDNA sequencing. The Colwellia strains are obligate psychrophiles, with a growth range restricted to 15 °C or less. As such, these organisms have less need for fluidity adaptation in the acyl moiety of the outer membrane, and they do not display alterations in lipid A based on growth temperature. Both Psychrobacter and Colwellia make use of extensive single-methylene variation in the size of their lipid A molecules. Such single-carbon variations in acyl size were thought to be restricted to psychrotolerant (facultative) species, but its presence in these Colwellia species shows that odd-chain acyl units and a single-carbon variation in lipid A structure are present in obligate psychrophiles, as well.

Coastal methane hydrate deposits are globally abundant. There is a need to understand the deep sediment sourced methane energy contribution to shallow sediment carbon relative to terrestrial sources and phytoplankton. Shallow sediment and porewater samples were collected from Atwater Valley, Texas-Louisiana Shelf, Gulf of Mexico near a seafloor mound feature identified in geophysical surveys as an elevated bottom seismic reflection. Geochemical data revealed off-mound methane diffusion and active fluid advection on-mound. Gas composition (average methane/ethane ratio ~11,000) and isotope ratios of methane on the mound (average δ13CCH4(g) = −71.2‰; D14CCH4(g) = −961‰) indicate a deep sediment, microbial source. Depleted sediment organic carbon values on mound (δ13CSOC = −25.8‰; D14CSOC = −930‰) relative to off-mound (δ13CSOC = −22.5‰; D14CSOC = −629‰) suggest deep sourced ancient carbon is incorporated into shallow sediment organic matter. Porewater and sediment data indicate inorganic carbon fixed during anaerobic oxidation of methane is a dominant contributor to on-mound shallow sediment organic carbon cycling. A simple stable carbon isotope mass balance suggests carbon fixation of dissolved inorganic carbon (DIC) associated with anaerobic oxidation of hydrate-sourced CH4 contributes up to 85% of shallow sediment organic carbon.

An autonomous surface vehicle instrumented with optical and acoustical sensors was deployed in Kane'ohe Bay, HI, U.S.A., to provide high-resolution, in situ observations of coral reef reflectance with minimal human presence. The data represented a wide range in bottom type, water depth, and illumination and supported more thorough investigations of remote sensing methods for identifying and mapping shallow reef features. The in situ data were used to compute spectral bottom reflectance and remote sensing reflectance, Rrs,λ, as a function of water depth and benthic features. The signals were used to distinguish between live coral and uncolonized sediment within the depth range of the measurements (2.5–5 m). In situRrs, λ were found to compare well with remotely sensed measurements from an imaging spectrometer, the Airborne Visible and Infrared Imaging Spectrometer (AVIRIS), deployed on an aircraft at high altitude. Cloud cover and in situ sensor orientation were found to have minimal impact on in situRrs, λ, suggesting that valid reflectance data may be collected using autonomous surveys even when atmospheric conditions are not favorable for remote sensing operations. The use of reflectance in the red and near infrared portions of the spectrum, expressed as the red edge height, REHλ, was investigated for detecting live aquatic vegetative biomass, including coral symbionts and turf algae. The REHλ signal from live coral was detected in Kane'ohe Bay to a depth of approximately 4 m with in situ measurements. A remote sensing algorithm based on the REHλ signal was defined and applied to AVIRIS imagery of the entire bay and was found to reveal areas of shallow, dense coral and algal cover. The peak wavelength of REHλ decreased with increasing water depth, indicating that a more complete examination of the red edge signal may potentially yield a remote sensing approach to simultaneously estimate vegetative biomass and bathymetry in shallow water.

In June 2007 sediment cores were collected in Alaminos Canyon, Gulf of Mexico across a series of seismic data profiles indicating rapid transitions between the presence of methane hydrates and vertical gas flux. Vertical profiles of dissolved sulfate, chloride, calcium, magnesium, and dissolved inorganic carbon (DIC) concentrations in porewaters, headspace methane, and solid phase carbonate concentrations were measured at each core location to investigate the cycling of methane-derived carbon in shallow sediments overlying the hydrate bearing strata. When integrated with stable carbon isotope ratios of DIC, geochemical results suggest a significant fraction of the methane flux at this site is cycled into the inorganic carbon pool. The incorporation of methane-derived carbon into dissolved and solid inorganic carbon phases represents a significant sink in local carbon cycling and plays a role in regulating the flux of methane to the overlying water column at Alaminos Canyon. Targeted, high-resolution geochemical characterization of the biogeochemical cycling of methane-derived carbon in shallow sediments overlying hydrate bearing strata like those in Alaminos Canyon is critical to quantifying methane flux and estimating methane hydrate distributions in gas hydrate bearing marine sediments.

Moderate elevated vertical methane (CH4) flux is associated with sediment accretion and raised fluid expulsion at the Hikurangi subduction margin, located along the northeast coast of New Zealand. This focused CH4 flux contributes to the cycling of inorganic and organic carbon in solid phase sediment and pore water. Along a 7 km offshore transect across the Porangahau Ridge, vertical CH4 flux rates range from 11.4 mmol·m−2·a−1 off the ridge to 82.6 mmol·m−2·a−1 at the ridge base. Stable carbon isotope ratios (δ13C) in pore water and sediment were variable across the ridge suggesting close proximity of heterogeneous carbon sources. Methane stable carbon isotope ratios ranging from −107.9‰ to −60.5‰ and a C1:C2 of 3000 indicate a microbial, or biogenic, source. Near ridge, average δ13C for pore water and sediment inorganic carbon were 13C-depleted (−28.7‰ and −7.9‰, respectively) relative to all core subsamples (−19.9‰ and −2.4‰, respectively) suggesting localized anaerobic CH4 oxidation and precipitation of authigenic carbonates. Through the transect there was low contribution from anaerobic oxidation of CH4 to organic carbon pools; for all cores δ13C values of pore water dissolved organic carbon and sediment organic carbon averaged −24.4‰ and −22.1‰, respectively. Anaerobic oxidation of CH4 contributed to pore water and sediment organic carbon near the ridge as evidenced by carbon isotope values as low as to −42.8‰ and −24.7‰, respectively. Carbon concentration and isotope analyses distinguished contributions from CH4 and phytodetrital carbon sources across the ridge and show a low methane contribution to organic carbon.

In September 2009, a series of sediment cores were collected across the Alaskan Beaufort Sea shelf-slope. Sediment and porewater organic carbon (OC) concentrations and stable carbon isotope ratios (δ13C) were measured to investigate spatial variations in sediment organic matter (OM) sources and distribution of these materials across the shelf. Cores were collected along three main nearshore (shelf) to offshore (slope) sampling lines (transects) from east-to-west along the North Slope of Alaska: Hammerhead (near Camden Bay), Thetis Island (near Prudhoe Bay), and Cape Halkett (towards Point Barrow). Measured sediment organic carbon (TOC) and porewater dissolved organic carbon (DOC) concentrations and their respective δ13C values were used to investigate the relative contribution of different OM sources to sediment OC pool cycled at each location. Sources of OM considered included: water column-sourced phytodetritus, deep sediment methane (CH4), and terrestrial, tundra/river-sourced OM. Results of these measurements, when coupled with results from previous research and additional analyses of sediment and porewater composition, show a pattern of spatial variation in sediment OC concentrations, OM source contributions, and OM cycled along the Alaskan Beaufort Sea shelf. In general, measured sediment total organic carbon (TOC) concentrations, δ13CTOC values, porewater DOC concentrations, and δ13CDOC values are consistent with an east-to-west transport of modern Holocene sediments with higher OC concentrations primarily sourced from relatively labile terrestrial, tundra OM sources and phytodetritus along the Alaskan Beaufort shelf. Sediment transport along the shelf results in the medium-to-long term accumulation and burial of sediment OM focused to the west which in turn results in higher biogenic CH4 production rates and higher upward CH4 diffusion through the sediments resulting in CH4−AMO-sourced contribution to sediment OC westward along the shelf. Understanding current OM sources and distributions along the Alaskan Beaufort shelf is important for enhancing models of carbon cycling in Arctic coastal shelf systems. This will help support the prediction of the climate response of the Arctic created in the face of future warming scenarios.

Abstract Background Although various treatments help reduce abdominal pain, real-world pain medication utilization among patients with Crohn’s disease (CD) or ulcerative colitis (UC) receiving advanced therapies is poorly understood. The aim is to understand the utilization of pain medication 12 months before and after the initiation of advanced therapies among patients with newly diagnosed CD or UC. Methods This retrospective, observational cohort study used administrative medical and pharmacy claims data of patients with CD or UC from HealthCore Integrated Research Database (HIRD®). The data from patients with use of pain medication over 12 months follow-up (after the initiation date of advanced therapies) were collected and analyzed. Differences in the use of pain medication 12 months before and after the initiation of advanced therapies were assessed using McNemar's and Wilcoxon signed-rank test. Results Prior to initiating advanced therapies, 23.1% of patients with CD (N = 540) received nonsteroidal anti-inflammatory drugs (NSAIDs), 78.1% glucocorticoids, 49.4% opioids, and 29.3% neuromodulators; similarly, 20.9% of patients with UC (N = 373) received NSAIDs, 91.4% glucocorticoids, 40.8% opioids, and 29.5% neuromodulators. After receiving advanced therapies for 12 months, patients reported a reduction in the use of steroids (78.1% vs. 58.9%, P

The molecular and cellular underpinnings of cribriform prostate cancer aggressiveness remain to be explored. Here, the authors perform single-cell RNA-sequencing, TCR sequencing and histology and reveal cancer cell intrinsic pathways and an immunosuppressive tumour microenvironment.

Abstract We parametrise the space of all possible flavour non-universal u $$ \mathfrak{u} $$ (1) X extensions of the Standard Model that embed inside anomaly-free semi-simple gauge theories, including up to three right-handed neutrinos. More generally, we parametrise all abelian extensions (i.e. by any number of u $$ \mathfrak{u} $$ (1)’s) of the SM with such semi-simple completions. The resulting space of abelian extensions is a collection of planes of dimensions ≤ 6. Numerically, we find that roughly 2.5% of anomaly-free u $$ \mathfrak{u} $$ (1) X extensions of the SM with a maximum charge ratio of ±10 can be embedded in such semi-simple gauge theories. Any vector-like anomaly-free abelian extension embeds (at least) inside g $$ \mathfrak{g} $$ = su $$ \mathfrak{su} $$ (12) ⊕ su $$ \mathfrak{su} $$ (2) L ⊕ su $$ \mathfrak{su} $$ (2) R . We also provide a simple computer program that tests whether a given u 1 X 1 $$ \mathfrak{u}{(1)}_{X^1} $$ ⊕ u 1 X 2 $$ \mathfrak{u}{(1)}_{X^2} $$ ⊕ . . . charge assignment has a semi-simple completion and, if it does, outputs a set of maximal gauge algebras in which the sm $$ \mathfrak{sm} $$ ⊕ u 1 X 1 $$ \mathfrak{u}{(1)}_{X^1} $$ ⊕ u 1 X 2 $$ \mathfrak{u}{(1)}_{X^2} $$ ⊕ . . . model may be embedded. We hope this is a useful tool in pointing the way from sm $$ \mathfrak{sm} $$ ⊕ u 1 X 1 $$ \mathfrak{u}{(1)}_{X^1} $$ ⊕ u 1 X 2 $$ \mathfrak{u}{(1)}_{X^2} $$ ⊕ . . . models, which have many phenomenological uses, to their unified gauge completions in the ultraviolet.

Abstract We propose that the electroweak and flavour quantum numbers of the Standard Model (SM) could be unified at high energies in an SU(4) × Sp(6) L × Sp(6) R anomaly-free gauge model. All the SM fermions are packaged into two fundamental fields, Ψ L ∼ (4 , 6 , 1) and Ψ R ∼ (4 , 1 , 6), thereby explaining the origin of three families of fermions. The SM Higgs, being electroweakly charged, necessarily becomes charged also under flavour when embedded in the UV model. It is therefore natural for its vacuum expectation value to couple only to the third family. The other components of the UV Higgs fields are presumed heavy. Extra scalars are needed to break this symmetry down to the SM, which can proceed via ‘flavour-deconstructed’ gauge groups; for instance, we propose a pattern Sp(6) L → ∏ i = 1 3 SU 2 L , i → SU 2 L $$ {\prod}_{i=1}^3\mathrm{SU}{(2)}_{L,i}\to \mathrm{SU}{(2)}_L $$ for the left-handed factor. When the heavy Higgs components are integrated out, realistic quark Yukawa couplings with in-built hierarchies are naturally generated without any further ingredients, if we assume the various symmetry breaking scalars condense at different scales. The CKM matrix that we compute is not a generic unitary matrix, but it can precisely fit the observed values.

Abstract Extensions of the minimal supersymmetric standard model (MSSM) gauge group abound in the literature. Several of these include an additional U(1) X gauge group. Chiral fermions’ charge assignments under U(1) X are constrained to cancel local anomalies in the extension and they determine the structure and phenomenology of it. We provide all anomaly-free charge assignments up to a maximum absolute charge of Q max = 10, assuming that the chiral superfield content of the model is that of the MSSM plus up to three Standard Model (SM) singlet superfields. The fermionic components of these SM singlets may play the rôle of right-handed neutrinos, whereas one of the scalar components may play the rôle of the flavon, spontaneously breaking U(1) X . Easily scanned lists of the charge assignments are made publicly available on Zenodo. For the case where no restriction is placed upon Q max, we also provide an analytic parameterisation of the general solution using simple techniques from algebraic geometry.

Abstract Background Antipsychotic prescribing to Medicaid-enrolled youth has been the target of numerous policy initiatives, including prior authorization and quality monitoring programs, which often target specific populations. Whether these efforts have changed the level or composition of antipsychotic prescribing is unclear. Methods Using 2014–2018 administrative claims data for Medicaid enrollees aged 21 years and under in Philadelphia, Pennsylvania, we measured antipsychotic prescription fills overall and for youth without an approved indication (autism, bipolar disorder, or psychosis). We then assessed whether trends differed for populations that have been targeted by policy initiatives, including younger children and foster care-enrolled youth. We also identified the most common approved and unapproved indications and examined whether the treatment duration of antipsychotic prescriptions differed based on whether the youth had an approved or unapproved indication. Results Overall, the number of Medicaid youth with an antipsychotic prescription fill halved between 2014 and 2018. Youth aged 17 years and under and foster care-enrolled youth, who were targeted by prior authorization and quality improvement efforts, experienced larger declines. Roughly half of prescriptions were for unapproved indications in both 2014 and 2018; the most common unapproved indication was ADHD, and the treatment duration was shorter for unapproved indications compared to approved indications. Conclusions Antipsychotic prescribing to Medicaid-enrolled youth is declining, particularly among populations that have been targeted by policy initiatives like prior authorization and quality monitoring programs. Despite the fact that these initiatives often assess diagnostic criteria, half of antipsychotic prescriptions were for unapproved indications in both 2014 and 2018. More research is needed to gauge whether this prescribing is appropriate.

Abstract Background Active surveillance (AS) is a management option for men diagnosed with lower risk prostate cancer. There is wide variation in all aspects of AS internationally, from patient selection to investigations and follow-up intervals, and a lack of clear evidence on the optimal approach to AS. This study aimed to provide guidance for clinicians from an international panel of prostate cancer experts. Methods A modified Delphi approach was undertaken, utilising two rounds of online questionnaires followed by a face-to-face workshop. Participants indicated their level of agreement with statements relating to patient selection for AS via online questionnaires on a 7-point Likert scale. Factors not achieving agreement were iteratively developed between the two rounds of questionnaires. Draft statements were presented at the face-to-face workshop for discussion and consensus building. Results 12 prostate cancer experts (9 urologists, 2 academics, 1 radiation oncologist) participated in this study from a range of geographical regions (4 USA, 4 Europe, 4 Australia). Complete agreement on statements presented to the participants was 29.4% after Round One and 69.0% after Round Two. Following robust discussions at the face-to-face workshop, agreement was reached on the remaining statements. PSA, PSA density, Multiparametric MRI, and systematic biopsy (with or without targeted biopsy) were identified as minimum diagnostic tests required upon which to select patients to recommend AS as a treatment option for prostate cancer. Patient factors and clinical parameters that identified patients appropriate to potentially receive AS were agreed. Genetic and genomic testing was not recommended for use in clinical decision-making regarding AS. Conclusions The lack of consistency in the practice of AS for men with lower risk prostate cancer between and within countries was reflected in this modified Delphi study. There are, however, areas of common practice and agreement from which clinicians practicing in the current environment can use to inform their clinical practice to achieve the best outcomes for patients.

Inherited alterations of the 8q24 genomic locus are linked to cancer etiology. Here, the authors investigate hereditary prostate cancer to reveal ancestral risk-altering haplotypes, the sentinel genetic variants optimally predicting risk, and causal mutation candidates.

Cerebral malaria (CM) affects children and adults, but brain swelling is more severe in children. To investigate features associated with brain swelling in malaria, we performed blood profiling and brain MRI in a cohort of pediatric and adult patients with CM in Rourkela, India, and compared them with an African pediatric CM cohort in Malawi. We determined that higher plasma Plasmodium falciparum histidine rich protein 2 (PfHRP2) levels and elevated var transcripts that encode for binding to endothelial protein C receptor (EPCR) were linked to CM at both sites. Machine learning models trained on the African pediatric cohort could classify brain swelling in Indian children CM cases but had weaker performance for adult classification, due to overall lower parasite var transcript levels in this age group and more severe thrombocytopenia in Rourkela adults. Subgrouping of patients with CM revealed higher parasite biomass linked to severe thrombocytopenia and higher Group A–EPCR var transcripts in mild thrombocytopenia. Overall, these findings provide evidence that higher parasite biomass and a subset of Group A–EPCR binding variants are common features in children and adult CM cases, despite age differences in brain swelling.

Cytoadhesion of Plasmodium falciparum-infected red blood cells is a virulence determinant associated with microvascular obstruction and organ complications. The gastrointestinal tract is a major site of sequestration in fatal cerebral malaria cases and kidney complications are common in severe malaria, but parasite interactions with these microvascular sites are poorly characterized. To study parasite tropism for different microvascular sites, we investigated binding of parasite lines to primary human microvascular endothelial cells from intestine (HIMEC) and peritubular kidney (HKMEC) sites. Of the three major host receptors for P. falciparum, CD36 had low or negligible expression; endothelial protein C receptor (EPCR) had the broadest constitutive expression; and intercellular adhesion molecule 1 (ICAM-1) was weakly expressed on resting cells and was strongly upregulated by TNF-α on primary endothelial cells from the brain, intestine, and peritubular kidney sites. By studying parasite lines expressing var genes linked to severe malaria, we provide evidence that both the DC8 and Group A EPCR-binding subsets of the P. falciparum erythrocyte membrane protein 1 (PfEMP1) family encodes binding affinity for brain, intestinal, and peritubular kidney endothelial cells, and that DC8 parasite adhesion was partially dependent on EPCR. Collectively, these findings raise the possibility of a brain-gut-kidney binding axis contributing to multi-organ complications in severe malaria.

Identifying patterns of organismal distribution can provide valuable insights for basic and applied marine and coastal ecology because understanding where animals are located is foundational to both research and science-based conservation. Understanding variation in distributional patterns can lead to a better assessment of ecological drivers and an improved ability to predict consequences of natural and altered relationships. Here, our purpose is to explore if quantifying coexisting groups of individual fish predators advances our understanding of field distribution patterns. Toward this end, we quantified locations of 59 acoustically tagged striped bass (Morone saxatilis) within a 26-stationary unit telemetry receiver array in Plum Island Estuary (PIE), MA, United States. We then used cluster analyses on spatial and temporal-spatial metrics from this dataset to (1) assess if distinct groups of individuals coexisted, (2) quantify group characteristics, and (3) test associations between groups and distribution (e.g., physical site type and region). Based on multiple lines of evidence, we identified four groups of striped bass with different space use patterns that persisted across seasons (summer and fall). Similar-sized striped bass clustered at spatial and temporal scales at which individuals within distinct groups could, and did, physically overlap. In addition, distributional groups were linked to components of physical site type and region suggesting that discrete groups of individuals can interact differently with the environment within the same ecological system. The identification of these distinct groups of individuals creates a baseline from which to explore further ecological implications of grouping behavior for research and conservation in geographically large, temporally dynamic, and spatially heterogeneous marine and coastal environments.

Issues related to safe and abundant energy production have been prominent in recent years. This is particularly true when society considers how to increase the quality of life by providing low-cost energy to citizens. A significant concern of the Gulf Cooperation Council (GCC) relates to the environmental effects of energy production and energy use associated with climate change. Efforts to reduce fossil fuel use and increase the use of renewable energy, together with the price volatility of fossil fuels, have seriously impacted the economics of many oil-producing countries, particularly the Gulf States, which has led to efforts to make their economies more diverse and less dependent on oil production. In this article, we introduce a multidisciplinary energy economics course developed at the Missouri University of Science and Technology that can be easily adopted by GCC universities to enhance future generations’ understanding of energy challenges.

Abstract Woody plant expansion into shrub and grasslands is a global and vexing ecological problem. In the Great Basin of North America, the expansion of pinyon–juniper (Pinus spp.–Juniperus spp.) woodlands is threatening the sagebrush (Artemisia spp.) biome. The Greater Sage‐grouse (Centrocercus urophasianus; sage‐grouse), a sagebrush obligate species, is widespread in the Great Basin and considered an indicator for the condition of sagebrush ecosystems. To assess the population response of sage‐grouse to landscape‐scale juniper removal, we analyzed a long‐term telemetry data set and lek counts with a Bayesian integrated population model in a before‐after‐control‐impact design. Population growth rates (λ) in a treatment area (Treatment) with juniper removal and a control area (Control) without juniper removal indicated the two areas generally experienced population increase, decrease, and stability in the same years. However, the difference in λ between study areas indicated a steady increase in the Treatment relative to the Control starting in 2013 (removals initiated in 2012), with differences of 0.13 and 0.11 in 2016 and 2017, respectively. Retrospective sensitivity analysis suggested the dynamics in λ were driven by increases in juvenile, adult, first nest, and yearling survival in the Treatment relative to the Control. These findings demonstrate the effectiveness of targeted conifer removal as a management strategy for conserving sage‐grouse populations in sagebrush steppe affected by conifer expansion. Examples of positive, population‐level responses to habitat management are exceptionally rare for terrestrial vertebrates, and this study provides promising evidence of active management that can be implemented to aid recovery of an imperiled species and biome.

Septic synovitis is a critical orthopedic condition in horses. Early intervention is key, with antibiotic therapy typically initiated prior to culture and susceptibility reports becoming available. The pharmacokinetics of several antibiotics have been studied in horses for use in intravenous regional limb perfusion (IVRLP) for septic synovitis, including the carbapenem antibiotic, meropenem. For a variety of factors, some veterinary clinicians may select IVRLP meropenem as therapy for these cases. Meropenem is a vital antibiotic in human medicine, making veterinary use divisive. However, verifying the efficacy of meropenem contrasted to other IVRLP antibiotics is essential for appropriate antimicrobial stewardship. To investigate this, equine patient medical records at a single veterinary teaching hospital were examined. Cases treated with meropenem or gentamicin via IVRLP for septic synovitis were retrospectively analyzed for demographics, diagnostics, treatments, outcomes, and adverse effects. Twenty-three meropenem and 37 gentamicin treated horses were analyzed; demographic information was similar between groups. In the meropenem group, nine horses received meropenem only; the remainder received another antibiotic initially then changed to meropenem. Structures infected included joints (meropenem = 13, gentamicin = 17), tendon sheaths (meropenem = 5, gentamicin = 8) and navicular bursae (meropenem = 2, gentamicin = 6). Overall survival to discharge was 86% (52/60), with meropenem 91% (21/23) and gentamicin 84% (31/37), with no statistically significant differences noted between meropenem or gentamicin groups for overall survival to discharge or outcome after discharge. Twenty-four of 26 bacterial isolates obtained from culture were reported as sensitive to imipenem, a carbapenem antibiotic similar to meropenem. Reported susceptibility to other antibiotics such as ceftiofur (n = 22/26), ampicillin (n = 18/26), amikacin (n = 15/26), or gentamicin (n = 12/26) was also frequently present. In the population of this study, antimicrobial activity augmented with IVRLP using either meropenem or gentamicin both appear to be an effective treatment for septic synovial structures, therefore, less critical antimicrobials may be a viable and more judicious treatment option.

Bestrophinopathies are a group of clinically distinct inherited retinal dystrophies that typically affect the macular region, an area synonymous with central high acuity vision. This spectrum of disorders is caused by mutations in bestrophin1 ( BEST1 ), a protein thought to act as a Ca 2+ -activated Cl - channel in the retinal pigment epithelium (RPE) of the eye. Although bestrophinopathies are rare, over 250 individual pathological mutations have been identified in the BEST1 gene, with many reported to have various clinical expressivity and incomplete penetrance. With no current clinical treatments available for patients with bestrophinopathies, understanding the role of BEST1 in cells and the pathological pathways underlying disease has become a priority. Induced pluripotent stem cell (iPSC) technology is helping to uncover disease mechanisms and develop treatments for RPE diseases, like bestrophinopathies. Here, we provide a comprehensive review of the pathophysiology of bestrophinopathies and highlight how patient-derived iPSC-RPE are being used to test new genomic therapies in vitro.

Cautery disbudding is a painful procedure performed on goat kids to prevent horn growth that may result in brain injury. Thermal damage to the cerebral cortex of the brain and subsequent neurologic disease is a primary concern. Cautery iron application time may affect transmission of heat to the brain; however, research in this area is scarce. Therefore, the objective of this pilot study was to evaluate the effect of iron application time on brain injury of goat kids. A total of six buck and doe kids

Abstract Much interest lies in the identification of manageable habitat variables that affect key vital rates for species of concern. For ground‐nesting birds, vegetation surrounding the nest may play an important role in mediating nest success by providing concealment from predators. Height of grasses surrounding the nest is thought to be a driver of nest survival in greater sage‐grouse (Centrocercus urophasianus; sage‐grouse), a species that has experienced widespread population declines throughout their range. However, a growing body of the literature has found that widely used field methods can produce misleading inference on the relationship between grass height and nest success. Specifically, it has been demonstrated that measuring concealment following nest fate (failure or hatch) introduces a temporal bias whereby successful nests are measured later in the season, on average, than failed nests. This sampling bias can produce inference suggesting a positive effect of grass height on nest survival, though the relationship arises due to the confounding effect of plant phenology, not an effect on predation risk. To test the generality of this finding for sage‐grouse, we reanalyzed existing datasets comprising >800 sage‐grouse nests from three independent studies across the range where there was a positive relationship found between grass height and nest survival, including two using methods now known to be biased. Correcting for phenology produced equivocal relationships between grass height and sage‐grouse nest survival. Viewed in total, evidence for a ubiquitous biological effect of grass height on sage‐grouse nest success across time and space is lacking. In light of these findings, a reevaluation of land management guidelines emphasizing specific grass height targets to promote nest success may be merited.

Porcine reproductive and respiratory syndrome viruses (PRRSV) are responsible for one of the most economically important diseases affecting the global pig industry. On-farm high-efficiency particulate air (HEPA) filtration systems can effectively reduce airborne transmission of PRRSV and the incidence of PRRS, but they are costly, and their adoption is limited. Therefore, there is a need for low-cost alternatives, such as antimicrobial filters impregnated with antiviral nanoparticles (AVNP). During the past 10 years, tailored intermetallic/multi-elemental AVNP compositions have demonstrated effective performance against human viruses. In this study, a panel of five AVNP was evaluated for viricidal activity against PRRSV. Three AVNP materials: AVNP2, copper nanoparticles (CuNP), and copper oxide nanoparticles (CuONP), were shown to exert a significant reduction (>99.99%) in virus titers at 1.0% (w/v) concentration. Among the three, CuNP was the most effective at lower concentrations. Further experiments revealed that AVNP generated significant reductions in viral titers within just 1.5 min. For an optimal reduction in viral titers, direct contact between viruses and AVNP was required. This was further explained by the inert nature of these AVNP, where only negligible leaching concentrations of Ag/Cu ions (0.06–4.06 ppm) were detected in AVNP supernatants. Real-time dynamic light scatting (DLS) and transmission electron microscopic (TEM) analyses suggested that the mono-dispersive hydrodynamic behavior of AVNPs may have enhanced their antiviral activity against PRRSV. Collectively, these data support the further evaluation of these AVNP as candidate nanoparticles for incorporation into antimicrobial air-filtration systems to reduce transmission of PRRSV and other airborne pathogens.

ABSTRACT Cerebral malaria is a severe neurological complication associated with sequestration of Plasmodium falciparum-infected erythrocytes (IE) in the brain microvasculature, but the specific binding interactions remain under debate. Here, we have generated an engineered three-dimensional (3D) human brain endothelial microvessel model and studied P. falciparum binding under the large range of physiological flow velocities that occur in both health and disease. Perfusion assays on 3D microvessels reveal previously unappreciated phenotypic heterogeneity in parasite binding to tumor necrosis factor alpha (TNF-α)-activated brain endothelial cells. While clonal parasite lines expressing a group B P. falciparum erythrocyte membrane protein 1 (PfEMP1) present an increase in binding to activated 3D microvessels, P. falciparum-IE expressing DC8-PfEMP1 present a decrease in binding. The differential response to endothelium activation is mediated by surface expression changes of endothelial protein C receptor (EPCR) and intercellular adhesion molecule 1 (ICAM-1). These findings demonstrate heterogeneity in parasite binding and provide evidence for a parasite strategy to adapt to a changing microvascular environment during infection. The engineered 3D human brain microvessel model provides new mechanistic insight into parasite binding and opens opportunities for further studies on malaria pathogenesis and parasite-vessel interactions. IMPORTANCE Cerebral malaria research has been hindered by the inaccessibility of the brain. Here, we have developed an engineered 3D human brain microvessel model that mimics the blood flow rates and architecture of small blood vessels to study how P. falciparum-infected human erythrocytes attach to brain endothelial cells. By studying parasite lines with different adhesive properties, we show that the malaria parasite binding rate is heterogeneous and strongly influenced by physiological differences in flow and whether the endothelium has been previously activated by TNF-α, a proinflammatory cytokine that is linked to malaria disease severity. We also show the importance of human EPCR and ICAM-1 in parasite binding. Our model sheds new light on how P. falciparum binds within brain microvessels and provides a powerful method for future investigations of recruitment of human brain pathogens to the blood vessel lining of the brain.

ABSTRACT The clinical presentation of severe Plasmodium falciparum malaria differs between children and adults, but the mechanistic basis for this remains unclear. Contributing factors to disease severity include total parasite biomass and the diverse cytoadhesive properties mediated by the polymorphic var gene parasite ligand family displayed on infected erythrocytes. To explore these factors, we performed a multicohort analysis of the contribution of var expression and parasite biomass to severe malaria in two previously published pediatric cohorts in Tanzania and Malawi and an adult cohort in India. Machine learning analysis revealed independent and complementary roles for var adhesion types and parasite biomass in adult and pediatric severe malaria and showed that similar var profiles, including upregulation of group A and DC8 var, predict severe malaria in adults and children. Among adults, patients with multiorgan complications presented infections with significantly higher parasite biomass without significant differences in var adhesion types. Conversely, pediatric patients with specific complications showed distinct var signatures. Cerebral malaria patients showed broadly increased expression of var genes, in particular group A and DC8 var, while children with severe malaria anemia were classified based on high transcription of DC8 var only. This study represents the first large multisite meta-analysis of var expression, and it demonstrates the presence of common var profiles in severe malaria patients of different ages across distant geographical sites, as well as syndrome-specific disease signatures. The complex associations between parasite biomass, var adhesion type, and clinical presentation revealed here represent the most comprehensive picture so far of the relationship between cytoadhesion, parasite load, and clinical syndrome. IMPORTANCE P. falciparum malaria can cause multiple disease complications that differ by patient age. Previous studies have attempted to address the roles of parasite adhesion and biomass in disease severity; however, these studies have been limited to single geographical sites, and there is limited understanding of how parasite adhesion and biomass interact to influence disease manifestations. In this meta-analysis, we compared parasite disease determinants in African children and Indian adults. This study demonstrates that parasite biomass and specific subsets of var genes are independently associated with detrimental outcomes in both childhood and adult malaria. We also explored how parasite var adhesion types and biomass play different roles in the development of specific severe malaria pathologies, including childhood cerebral malaria and multiorgan complications in adults. This work represents the largest study to date of the role of both var adhesion types and biomass in severe malaria.

ABSTRACT The small Tim proteins belong to a group of mitochondrial intermembrane space chaperones that aid in the import of mitochondrial inner membrane proteins with internal targeting signals. Trypanosoma brucei, the protozoan parasite that causes African trypanosomiasis, possesses multiple small Tim proteins that include homologues of T. brucei Tim9 (TbTim9) and Tim10 (TbTim10) and a unique small Tim that shares homology with both Tim8 and Tim13 (TbTim8/13). Here, we found that these three small TbTims are expressed as soluble mitochondrial intermembrane space proteins. Coimmunoprecipitation and mass spectrometry analysis showed that the small TbTims stably associated with each other and with TbTim17, the major component of the mitochondrial inner membrane translocase in T. brucei. Yeast two-hybrid analysis indicated direct interactions among the small TbTims; however, their interaction patterns appeared to be different from those of their counterparts in yeast and humans. Knockdown of the small TbTims reduced cell growth and decreased the steady-state level of TbTim17 and T. brucei ADP/ATP carrier (TbAAC), two polytopic mitochondrial inner membrane proteins. Knockdown of small TbTims also reduced the matured complexes of TbTim17 in mitochondria. Depletion of any of the small TbTims reduced TbTim17 import moderately but greatly hampered the stability of the TbTim17 complexes in T. brucei. Altogether, our results revealed that TbTim9, TbTim10, and TbTim8/13 interact with each other, associate with TbTim17, and play a crucial role in the integrity and maintenance of the levels of TbTim17 complexes. IMPORTANCE Trypanosoma brucei is the causative agent of African sleeping sickness. The parasite’s mitochondrion represents a useful source for potential chemotherapeutic targets. Similarly to yeast and humans, mitochondrial functions depend on the import of proteins that are encoded in the nucleus and made in the cytosol. Even though the machinery involved in this mitochondrial protein import process is becoming clearer in T. brucei, a comprehensive picture of protein complex composition and function is still lacking. In this study, we characterized three T. brucei small Tim proteins, TbTim9, TbTim10, and TbTim8/13. Although the parasite does not have the classical TIM22 complex that imports mitochondrial inner membrane proteins containing internal targeting signals in yeast or humans, we found that these small TbTims associate with TbTim17, the major subunit of the TbTIM complex in T. brucei, and play an essential role in the stability of the TbTim17 complexes. Therefore, these divergent proteins are critical for mitochondrial protein biogenesis in T. brucei.

ABSTRACT Plasmodium falciparum, the most deadly of the human malaria parasites, is a member of the Laverania subgenus that also infects African Great Apes. The virulence of P. falciparum is related to cytoadhesion of infected erythrocytes in microvasculature, but the origin of dangerous parasite adhesion traits is poorly understood. To investigate the evolutionary history of the P. falciparum cytoadhesion pathogenicity determinant, we studied adhesion domains from the chimpanzee malaria parasite P. reichenowi. We demonstrate that the P. reichenowi var gene repertoire encodes cysteine-rich interdomain region (CIDR) domains which bind human CD36 and endothelial protein C receptor (EPCR) with the same levels of affinity and at binding sites similar to those bound by P. falciparum. Moreover, P. reichenowi domains interfere with the protective function of the activated protein C-EPCR pathway on endothelial cells, a presumptive virulence trait in humans. These findings provide evidence for ancient evolutionary origins of two key cytoadhesion properties of P. falciparum that contribute to human infection and pathogenicity. IMPORTANCE Cytoadhesion of P. falciparum-infected erythrocytes in the microcirculation is a major virulence determinant. P. falciparum is descended from a subgenus of parasites that also infect chimpanzees and gorillas and exhibits strict host species specificity. Despite their high genetic similarity to P. falciparum, it is unknown whether ape parasites encode adhesion properties similar to those of P. falciparum or are as virulent in their natural hosts. Consequently, it has been unclear when virulent adhesion traits arose in P. falciparum and how long they have been present in the parasite population. It is also unknown whether cytoadhesive interactions pose a barrier to cross-species transmission. We show that parasite domains from the chimpanzee malaria parasite P. reichenowi bind human receptors with specificity similar to that of P. falciparum. Our findings suggest that parasite adhesion traits associated with both mild and severe malaria have much earlier origins than previously appreciated and have important implications for virulence evolution in a major human pathogen.

ABSTRACT Plasmodium falciparum malaria remains one of the most deadly infections worldwide. The pathogenesis of the infection results from the sequestration of infected erythrocytes (IRBC) in vital organs, including the brain, with resulting impairment of blood flow, hypoxia, and lactic acidosis. Sequestration occurs through the adhesion of IRBC to host receptors on microvascular endothelium by Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), a large family of variant surface antigens, each with up to seven extracellular domains that can bind to multiple host receptors. Consequently, antiadhesive therapies directed at single endothelial adhesion molecules may not be effective. In this study, we demonstrated that the serine protease thrombin, which is pivotal in the activation of the coagulation cascade, cleaved the major parasite adhesin on the surface of IRBC. As a result, adhesion under flow was dramatically reduced, and already adherent IRBC were detached. Thrombin cleavage sites were mapped to the Duffy binding-like δ1 (DBLδ1) domain and interdomains 1 and 2 in the PfEMP1 of the parasite line IT4var19. Furthermore, we observed an inverse correlation between the presence of thrombin and IRBC in cerebral malaria autopsies of children. We investigated a modified (R67A) thrombin and thrombin inhibitor, hirugen, both of which inhibit the binding of substrates to exosite I, thereby reducing its proinflammatory properties. Both approaches reduced the barrier dysfunction induced by thrombin without affecting its proteolytic activity on PfEMP1, raising the possibility that thrombin cleavage of variant PfEMP1 may be exploited as a broadly inhibitory antiadhesive therapy. IMPORTANCE Plasmodium falciparum malaria is the third leading cause of mortality due to a pathogen, with 214 million people infected and 438,000 deaths annually. The adhesion of Plasmodium falciparum-infected erythrocytes (IRBC) to microvascular endothelium is a major pathological process in severe malaria. While the recent implementation of artemisinin-based antimalarial therapy for severe malaria improves patient survival by targeting all parasite stages, antiparasite drugs alone may not immediately reverse pathophysiological processes in occluded vessels. Here we show that thrombin, an enzyme intimately involved in the clotting process, cleaves the main parasite adhesin expressed on the surface of IRBC, thereby preventing and reversing the binding of IRBC to endothelial cells. This beneficial effect of thrombin can be achieved by modified thrombins that cause significantly less clotting and vessel leakage while preserving the ability to cleave the parasite protein. Our results provide the basis for using modified thrombins as adjunctive therapy in severe malaria.

ABSTRACT Intercellular adhesion molecule 1 (ICAM-1) and the endothelial protein C receptor (EPCR) are candidate receptors for the deadly complication cerebral malaria. However, it remains unclear if Plasmodium falciparum parasites with dual binding specificity are involved in cytoadhesion or different parasite subpopulations bind in brain microvessels. Here, we investigated this issue by studying different subtypes of ICAM-1-binding parasite lines. We show that two parasite lines expressing domain cassette 13 (DC13) of the P. falciparum erythrocyte membrane protein 1 (PfEMP1) family have dual binding specificity for EPCR and ICAM-1 and further mapped ICAM-1 binding to the first DBLβ domain following the PfEMP1 head structure in both proteins. As PfEMP1 head structures have diverged between group A (EPCR binders) and groups B and C (CD36 binders), we also investigated how ICAM-1-binding parasites with different coreceptor binding traits influence P. falciparum-infected erythrocyte binding to endothelial cells. Whereas levels of binding to tumor necrosis factor alpha (TNF-α)-stimulated endothelial cells from the lung and brain by all ICAM-1-binding parasite lines increased, group A (EPCR and ICAM-1) was less dependent than group B (CD36 and ICAM-1) on ICAM-1 upregulation. Furthermore, both group A DC13 parasite lines had higher binding levels to brain endothelial cells (a microvascular niche with limited CD36 expression). This study shows that ICAM-1 is a coreceptor for a subset of EPCR-binding parasites and provides the first evidence of how EPCR and ICAM-1 interact to mediate parasite binding to both resting and TNF-α-activated primary brain and lung endothelial cells. IMPORTANCE Cerebral malaria is a severe neurological complication of P. falciparum infection associated with infected erythrocyte (IE) binding in cerebral vessels. Yet little is known about the mechanisms by which parasites adhere in the brain or other microvascular sites. Here, we studied parasite lines expressing group A DC13-containing PfEMP1 variants, a subset that has previously been shown to have high brain cell- and other endothelial cell-binding activities. We show that DC13-containing PfEMP1 variants have dual EPCR- and ICAM-1-binding activities and that both receptors are involved in parasite adherence to lung and brain endothelial cells. As both EPCR and ICAM-1 are implicated in cerebral malaria, these findings suggest the possibility that parasites with dual binding activities are involved in parasite sequestration to microvascular beds with low CD36 expression, such as the brain, and we urge more research into the multiadhesive properties of PfEMP1 variants.