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Arctic heat and freshwater budgets are highly sensitive to volume transports through Arctic-Subarctic straits. Here we investigate how the volume transports through these straits adjust to each other to maintain a mass balance in the Arctic on annual timescales. To this end, we use three models; two coupled global climate models, one with a third-degree horizontal ocean resolution (HiGEM1.1) and one with a twelfth-degree horizontal ocean resolution (HadGEM3), and one ocean-only model with an idealized polar basin (tenth-degree horizontal resolution). The two global climate models indicate that there is a strong anti-correlation between the Bering Strait throughflow and the transport through the Nordic Seas, a second strong anti-correlation between the transport through the Canadian Artic Archipelago (CAA) and the Nordic Seas transport, and a third strong anti-correlation between the Fram Strait and the Barents Sea throughflows. We find that part of the strait correlations is due to the strait transports being coincidentally driven by large-scale atmospheric forcing patterns such as the Arctic Oscillation. However, there is also a role for fast wave adjustments of some straits flows to perturbations in other straits since atmospheric forcing of individual strait flows alone cannot lead to near mass balance fortuitously every year. Idealized experiments with an ocean model (NEMO3.6) that investigate such causal strait relations suggest that perturbations in the Bering Strait are compensated preferentially in the Fram Strait due to the narrowness of the western Arctic shelf and the deeper depth of the Fram Strait.

Sea surface temperature (SST) seasonal extrema are important for water mass formation, intensification of tropical cyclones and coral bleaching, so should be well-represented in models used for future climate projections. Typically, climate model evaluations focus on annual or longer-term mean SST. However, accurate mean SST does not guarantee accurate seasonal extrema or annual cycles. Models that have no biases in mean SST can have biases in seasonal extrema and annual cycles, and vice versa.Here we assess seasonal extrema in a selection of CMIP6 model historical runs (including BCC-CSM2-MR, CanESM5, CESM2, GFDL-CM4 and GISS-E2-1-G), averaged over 1981-2010, against the World Ocean Atlas (WOA18) observational climatology. The magnitude and pattern of SST biases for seasonal extrema vary from model to model. GFDL-CM4 and CESM2 simulate SST extrema reasonably well, while BCC-CSM2-MR and GISS-E2-1-G have obvious deficiencies. The global area-weighted root mean square (RMS) difference from WOA18 is larger than 2oC in BCC-CSM2-MR and GISS-E2-1-G, and their common maximum bias (larger than 5oC) is the cold bias located in the subpolar North Pacific, Greenland Sea and Norwegian Sea. The model biases of maximum SST (summer SST) and minimum SST (winter SST) are in some cases different, leading to biased SST annual cycles. The SST biases are typically smaller for summer, except for models with significant winter cold bias in the high latitudes of the Northern Hemisphere (BCC-CSM2-MR and GISS-E2-1-G). Generally speaking, the bias of the SST annual cycle is smaller than that of seasonal extrema; models that are too cold in winter are typically also too cold in summer. In eastern boundary regions, the models have too small annual cycles. In these regions, the warm bias of winter SST is less than the warm bias of summer SST. This is because the warm bias in models due to poorly captured stratocumulus can be compensated by coastal upwelling, which cools the sea surface more in summer than in winter.We note that extra attention should be paid when evaluating SST extrema in some polar areas as the observational climatology there can be unrealistic, particularly in winter.

The Nordic Seas have a significant impact on global climate due to their role in providing dense overflows to the North Atlantic Ocean. However, the dramatic loss of sea ice in recent decades is creating a new atmosphere-ice-ocean environment where large swathes of the ocean that were previously ice-covered are now exposed to the atmosphere. Despite the largest sea-ice loss occurring in summer and autumn, the sea-ice loss in winter and spring is arguably more important for the climate system. Atmosphere-ocean coupling is the most intense in the extended winter, when convective mixing leads to water-mass modification processes, impacting the densest waters of the Atlantic Meridional Overturning Circulation. Here we focus on the marginal-ice-zone of the Nordic Seas where the air-sea temperature difference is large, promoting high heat flux events during periods of off-ice winds. We use both transient and control simulations of the coupled climate model HiGEM, which allows us to isolate the climate change response from the sea-ice retreat response. We find that wintertime sea-ice retreat leads to remarkable changes in ocean surface heat exchanges and wind energy input. As the sea ice edge retreats towards the Greenland coastline, there is a band of exposed ocean which was previously covered by ice. This exposure allows enhanced mechanical mixing by the wind and a greater loss of buoyancy from the ocean leading to deeper vertical mixing in the upper ocean. Sensible and latent heat fluxes from the ocean to the atmosphere provide the greatest loss of buoyancy. However, climate warming inhibits this process as the atmosphere warms more rapidly than the ocean which reduces the sea-air temperature difference. Further away from the retreating ice edge, toward the centre of the Greenland Sea, the upper ocean warms, resulting in a more stratified water column. As a consequence, the depth of convective mixing reduces over the deep ocean and increases over shallower regions close to the coast. This leads to changes in the formation and properties of some of the water masses that enter the North Atlantic and thus may modify the ocean circulation in the subpolar seas in response to sea-ice decline.

A Portuguese isolate of Aspergillus fumigatus was found to contain three double-stranded (ds) RNA elements ranging in size from 1.1 to 1.8 kbp and comprising the genome of a strain of Aspergillus fumigatus partitivirus 1 (AfuPV-1) previously thought to contain only the two largest dsRNA elements. The sequence of the smallest dsRNA element is described here, completing the sequence of the AfuPV-1 genome. Sequence analysis of the element revealed an open reading frame encoding a protein of unknown function similar in size and distantly related to elements previously identified in other members of the family Partitiviridae.

Filamentous basidiomycetes are uncommon agents of human diseases, despite their ubiquitous presence in the environment. We present a case of symptomatic pulmonary infection in a 38-year-old male with cough and fever; a thin-walled cyst in the posterior left upper pulmonary lobe was revealed by radiography. A non-sporulating fungus was isolated from sputum and biopsy material from the cyst. ITS and LSU sequences placed the fungus phylogenetically in Agaricales, family Cyphellaceae, and identified it as a member of shelf fungi in Gloeostereum, but without identity to any known species. The new species is described as Gloeostereum cimri. The clinical strain showed high MIC to voriconazole (>8 µg/ml) but had low MIC to amphotericin B (0.5 µg/ml).

A sustainable future for sheep and goat production throughout the world is that derived from pastures and forages, grazed in situ, utilizing resources such as hill lands that are unable to support other productive enterprises. Providing appropriate nutrition, especially from non-human edible supplemental ingredients, to achieve production and profitability goals of the landholder can be challenging in these situations. This chapter uses case studies from New Zealand and the United States of America to highlight productivity and efficiency improvements over the past 30 years, and challenges associated with balancing the needs of production, biodiversity and urbanization, as a template to highlight the practical changes of forage use, management techniques and nutritional practices that have underpinned those changes. The principles highlighted may act as a blueprint for future efficiency gains and reduction of the environmental footprint of animal agriculture systems using pasture and forage-based nutrition systems.

Context Since COVID-19 (SARS-CoV-2) was first identified in the human population, it has had immediate and significant effects on peoples' health and the worldwide economy. In the absence of a vaccine, control of the virus involved limiting its spread through restrictions in the movement of people, goods and services. This has led to unprecedented impacts on labour availability, provision of goods and services, value chains, and markets. Objective Against the backdrop of COVID-19 control measures, this article summarises quantitative and qualitative assessments of the impacts, adaptations, and opportunities to increase the resilience of the agricultural systems in Australia and New Zealand. Methods Using both survey and interview methodologies, we describe the various agri-food systems and the impacts of the COVID-19 control measures across different industries, and discuss the results applying a resilience framework. Results As essential services, all agricultural activities except for fibre production have been permitted to continue during quarantine periods but have been exposed to the major flow-on effects of movement control. We found that, to June 2020, the impacts of the COVID-19 control measures on the agri-food sectors in both Australia and New Zealand have been relatively small and that this has been due to the high levels of resilience in the agricultural systems and the people running them. Conclusions We consider agri-food systems to be comprised of multiple subsystems with varying vulnerability to external influences. Agri-food systems were resilient to June 2020 at least, and that resilience was achieved via one or more subsystems that were able to compensate for the more vulnerable subsystems. We contrast the resilience of industries that have high plasticity (that can have a flow of material that can safely vary in time) to more rigid industries that are dependent on a steady flow of material with little or no storage. Ultimately both types of industries were resilient, but they achieved that resilience via compensating subsystems. High plasticity industries relied on their production and processing subsystem; rigid industries engaged their institutional subsystem to achieve the same end. The social and cultural subsystem was important across all industries. Significance It is not yet clear if the current resilience mechanisms can persist under the continued onslaught of the virus. We indicate the need to capture longer term effects and analysis during the more sustained effects of the virus and through a recovery period. We anticipate a follow-up study in 2022.

We describe the successful treatment of a series of 30 zoonotic sporotrichosis cases from southern Brazil. Sporothrix brasiliensis was the species genotypically identified in all 25 confirmed cases. Five other cases were classified as probable, without laboratory confirmation, but with clinical and epidemiological data of cat-transmitted sporotrichosis. Two isolates were sequenced by translation elongation factor-1 alpha (EF1α) loci in order to compare their sequences, and both of them showed distinct genotypes from S. brasiliensis strains from other Brazilian states. Itraconazole (ITZ) or potassium iodide (KI) were the first choice treatment in 28 and 2 cases, respectively. Microdilution assay showed a wild-type profile of S. brasiliensis isolates to ITZ. However, a lack of clinical response occurred in 42% of cases, especially those treated with ITZ 100 mg/day, and treatment needed modifications, by either increased doses or antifungal combinations. Clinical cure required a mean of 187 days of treatment, which was dependent on the clinical form of the disease and age of patients. Therapy, including dosages and durations, for cutaneous forms of sporotrichosis requires re-evaluation, since cases caused by S. brasiliensis may influence treatment efficacy.

Introduction Cryptococcus neoformans is an opportunistic pathogen that causes ∼15% mortality in AIDS patients. Rio Grande City, Rio Grande do Sul (RS), Brazil, has the highest national rate of HIV/AIDS, considering cities with population more than 100,000 habitants. Objective We aimed to evaluate the clinical and epidemiological profile of cryptococcosis in a reference service for HIV-AIDS patients in the South region of Brazil, over seven years. Material and methods A retrospective study was performed including all cryptococcosis cases diagnosed at the University Hospital, Federal University of Rio Grande (UH-FURG) between January 2010 and December 2016. Results Seventy cases of cryptococcosis were diagnosis from 2010 to 2016 in the UH-FURG in the seven years of the study. These numbers were responsible for 2.1% to 8.1% of the hospitalizations/year for HIV patients. All were caused by C. neoformans infection (95% C. neoformans var. grubii VNI and 5% C. neoformans var. grubii VNII). Neurocryptococcosis was the major clinical manifestation and cryptococcosis was the HIV- defining condition in 40% of patients. The period of hospitalization was an average of 39.3 days (SD = 31.3), and more than half of patients (53%; 37/70) died after a mean of 82 days. Discussion The present study showed the importance of cryptococcosis as an AIDS-defining disease in HIV-AIDS patients in a tertiary hospital from Southern Brazil. More investment is necessary to reduce the impact of this opportunistic mycosis in HIV-AIDS patients from southern Brazil.

In the Southern Ocean the Antarctic Circumpolar Current is significantly steered by large topographic features, and subpolar gyres form in their lee. The geometry of topographic features in the Southern Ocean is highly variable, but the influence of this variation on the large-scale flow is poorly understood. Using idealized barotropic simulations of a zonal channel with a meridional ridge, it is found that the ridge geometry is important for determining the net zonal volume transport. A relationship is observed between ridge width and volume transport that is determined by the form stress generated by the ridge. Gyre formation is also highly reliant on the ridge geometry. A steep ridge allows gyres to form within regions of unblocked geostrophic (f/H) contours, with an increase in gyre strength as the ridge width is reduced. These relationships among ridge width, gyre strength, and net zonal volume transport emerge to simultaneously satisfy the conservation of momentum and vorticity.

AbstractThis study evaluated the effect of early weaning (EW) of artificially reared lambs using a restricted milk replacer (MR) feeding and step-down weaning system on the short- and long-term effects on growth, feed intake, selected blood metabolites and hormones, body composition, and small intestine development. Mixed-sex twin-born 2 to 5 d old lambs were randomly allocated to individual pens and fed MR at 20% of initial individual BW in week 1 and 15% in week 2 followed by weaning off MR by the end of week 4 (EW; n = 16) or week 6 (Control; Ctrl, n = 16) using a step-down procedure. Concentrate starter and fiber diets were offered ad libitum to week 9, then gradually removed over a 10-d period. All lambs were managed as a single group on pasture from weeks 6 to 16 of the trial. Feed intake was recorded daily in the first 6 wk, and BWs recorded weekly. At weeks 2, 4, 6, and 8, and pre- and postclostridial vaccination at week 8, blood samples were collected for analysis of selected blood metabolites, IGF-1, and immune function. Body composition was evaluated in eight animals per group at weeks 4 and 16 after euthanasia, and duodenal samples collected for histomorphometric evaluation. Early weaned lambs had lower DM, ME, CP, and NDF intake than Ctrl lambs at 21, 15, 21, and 36 d of rearing, respectively (P < 0.001), driven by lower intakes of MR from day 15 (P < 0.001) as per the experimental design, and lower total DMI of fiber (P = 0.001) from 21 to 42 d of rearing. Lamb BW tended (P = 0.097) to be lower in EW than Ctrl lambs from 5 to 10 wk of rearing, with lower ADG in EW lambs from weeks 3 to 6 (P = 0.041). Early weaning had negligible effects on duodenal morphology, organ, and carcass weights at weeks 4 and 16. Plasma metabolites (urea nitrogen, triglycerides, NEFA, glucose, and total protein) were similar between groups, while β-hydroxybutyrate was greater in EW than Ctrl lambs at weeks 4 and 6 (P = 0.018) but not week 8 indicative of early rumen development. Serum IGF-1 tended to be lower in EW than Ctrl lambs from weeks 2 to 6 only (P = 0.065). All lambs developed antibody responses postvaccination and there was no effect of treatment (P = 0.528). The results of this study illustrate that artificially reared lambs can be weaned off MR by 4 or 6 wk of rearing without compromising growth, small intestine morphology, major organ development, and body composition, nor immune function at either 4 (preweaning) or 16 (postweaning) wk of age.

Studies of cerebral haemodynamics have shown changes with increased exercise intensity, but the patterns have been highly variable and reliable associations with cognitive performance have not been identified. The aim of this study was to examine whether exercise-induced changes in oxygenated haemoglobin (O2Hb) led to changes in concomitant cognitive performance. This study examined cardiorespiratory and cerebral haemodynamics during multi-stage exercise from rest to exhaustion, with (Ex + C) and without (Ex) concurrent cognitive performance (Go/No-go task). The presence of the cognitive task affected both cardiorespiratory and cerebral haemodynamics. The patterns in the cerebral haemodynamics during Ex and Ex + C diverged above the respiratory compensation threshold (RCT), but differences were significant only at 100% $$\dot {V}{{\text{O}}_{{\text{2max}}}}$$ , displaying increased deoxygenated haemoglobin (HHb), decreased difference between oxygenated and deoxygenated haemoglobin (HbDiff), and decreased cerebral oxygenation (COx) during Ex + C. More complex haemodynamic trends against intensity during Ex + C suggested that the presence of a cognitive task increases cerebral metabolic demand at high exercise intensities. The levels of O2Hb, HHb, HbDiff and total haemoglobin increased most steeply at intensities around the RCT during both Ex and Ex + C, but these changes were not accompanied by improved cognitive performance. The primary hypothesis, that cognitive performance would match changes in O2Hb, was not supported. Small variations in reaction time and response accuracy across exercise intensities were not significant, suggesting that cognitive performance is unaffected by intense short-duration exercise. Our results add further evidence that exercise-induced changes in cerebral haemodynamics do not affect cognitive performance.

The enzyme human DNA polymerase η (Pol η) is critical for bypassing lesions during DNA replication. In addition to the two Mg(2+) ions aligning the active site, experiments suggest that a third Mg(2+) ion could play an essential catalytic role. Herein the role of this third metal ion is investigated with quantum mechanical/molecular mechanical (QM/MM) free energy simulations of the phosphoryl transfer reaction and a proposed self-activating proton transfer from the incoming nucleotide to the pyrophosphate leaving group. The simulations with only two metal ions in the active site support a sequential mechanism, with phosphoryl transfer followed by relatively fast proton transfer. The simulations with three metal ions in the active site suggest that the third metal ion may play a catalytic role through electrostatic interactions with the leaving group. These electrostatic interactions stabilize the product, making the phosphoryl transfer reaction more thermodynamically favorable with a lower free energy barrier relative to the activated state corresponding to the deprotonated 3’OH nucleophile, and also inhibit the subsequent proton transfer.

A novel series of EP4 agonists and antagonists have been identified, and then used to validate their potential in the treatment of inflammatory pain. This paper describes these novel ligands and their activity within a number of pre-clinical models of pain, ultimately leading to the identification of the EP4 partial agonist GSK726701A.

Invasive pulmonary aspergillosis (IPA) is classically considered an illness of severely immunocompromised patients with limited host defenses. However, IPA has been reported in immunocompetent but critically ill patients. This report describes two fatal cases of pathologically confirmed IPA in patients with influenza in the intensive care unit. One patient had influenza B infection, whereas the other had influenza A H1N1. Both patients died despite broad-spectrum antimicrobials, mechanical ventilation, and vasopressor support. Microscopic and histologic postmortem examination confirmed IPA. Review of the English language and foreign literature indicates that galactomannan antigen testing and classic radiologic findings for IPA may not be reliable in immunocompetent patients. Respiratory cultures which grow Aspergillus species in critically ill patients, particularly those with underlying influenza infection, should not necessarily be disregarded as contaminants or colonizers. Further research is needed to better understand the immunological relationship between influenza and IPA for improved prevention and treatment of influenza and Aspergillus co-infections.

The HIV-1 regulatory protein, trans-activator of transcription (Tat), interacts with opioids to potentiate neuroinflammation and neurodegeneration within the CNS. These effects may involve the C-C chemokine receptor type 5 (CCR5); however, the behavioral contribution of CCR5 on Tat/opioid interactions is not known. Using a transgenic murine model that expresses HIV-1 Tat protein in a GFAP-regulated, doxycycline-inducible manner, we assessed morphine tolerance, dependence, and reward. To assess the influence of CCR5 on these effects, mice were pretreated with oral vehicle or the CCR5 antagonist, maraviroc, prior to morphine administration. We found that HIV-1 Tat expression significantly attenuated the antinociceptive potency of acute morphine (2–64 mg/kg, i.p.) in non-tolerant mice. Consistent with this, Tat attenuated withdrawal symptoms among morphine-tolerant mice. Pretreatment with maraviroc blocked the effects of Tat, reinstating morphine potency in non-tolerant mice and restoring withdrawal symptomology in morphine-tolerant mice. Twenty-four hours following morphine administration, HIV-1 Tat significantly potentiated (∼3.5-fold) morphine-conditioned place preference and maraviroc further potentiated these effects (∼5.7-fold). Maraviroc exerted no measurable behavioral effects on its own. Protein array analyses revealed only minor changes to cytokine profiles when morphine was administered acutely or repeatedly; however, 24 h post morphine administration, the expression of several cytokines was greatly increased, including endogenous CCR5 chemokine ligands (CCL3, CCL4, and CCL5), as well as CCL2. Tat further elevated levels of several cytokines and maraviroc pretreatment attenuated these effects. These data demonstrate that CCR5 mediates key aspects of HIV-1 Tat-induced alterations in the antinociceptive potency and rewarding properties of opioids.

Herbage accumulation (HA) data from four sites were combined to (1) address the hypothesis that adjustments to the New Zealand Forage Value Index to account for grass–clover interactions are not required and (2) explore the main effects of clover and nitrogen (N) fertiliser treatments on pasture productivity. In summer and autumn, pastures based on mid-season heading ryegrass cultivars consistently supported a greater clover content than pastures based on late heading cultivars. However, the clover × cultivar interaction was significant in only 2 of 17 HA data sets available for analysis. Thus, the hypothesis is supported. Compared with ryegrass monocultures, mixtures yielded an additional 2.4 (in the Low-N treatment) and 1.2 (High-N treatment) t DM/ha/year. Apparent N transfer from clover to grass was 79 (Low N) and 27 (High N) kg N/ha/year, derived from estimated N fixation efficiencies of 66 (Low N) and 50 (High N) kg N/t clover DM grown.

Concentrations of nitrogen, phosphorus, potassium, calcium, magnesium, sulphur and sodium were measured in eight perennial ryegrass cultivars grown at four sites in New Zealand (Waikato, Manawatu, Canterbury and Southland). For the macronutrient analyses, herbage samples were collected from pure ryegrass plots, grown with two nitrogen fertiliser rates, with three sites sampled in autumn (April/May) and all four sites in spring (October/November) of 2013. There were significant differences among the cultivars for all elements except potassium, extending the results of earlier research to contemporary commercial cultivars. It is suggested that breeding ryegrass for lowered shoot phosphorus concentration should be prioritised because of the economic and environmental benefits to be gained from farming with reduced phosphate fertiliser requirements.

Item does not contain fulltext Aspergillosis is a fungal infection that primarily affects the respiratory tract. Amphotericin B has broad antifungal activity and is commonly used to treat aspergillosis, a fungal pneumonia that is a common sequela in oiled waterfowl as well as other birds in wildlife rehabilitation. Pharmacokinetic parameters of nebulized amphotericin B in an avian model have been reported, but those of direct intratracheal delivery have yet to be established. The objective of this study was to evaluate if a single 3 mg/kg dose of liposomal amphotericin B delivered intratracheally using a commercial atomizer would achieve plasma and lung tissue concentrations exceeding targeted minimum inhibitory concentrations (MIC) for Aspergillus species in adult mallard ducks (Anas platyrhynchos). Following intratracheal delivery, amphotericin B was present in lung parenchyma at concentrations above the targeted MIC of 1 mug/g for up to 9 days post-administration; however, distribution of the drug was uneven, with the majority of the drug concentrated in one lung lobe. Concentrations in the contralateral lung lobe and the kidneys were above the targeted MIC 1 day after administration but declined exponentially with a half-life of approximately 2 days. Plasma concentrations were never above the targeted MIC. Histological examination of the trachea, bronchi, lungs, heart, liver, and kidneys did not reveal any toxic changes. Using a commercial atomizer, intratracheal delivery of amphotericin B at 3 mg/kg resulted in lung parenchyma concentrations above 1 mug/ml with no discernable systemic effects. Further studies to establish a system of drug delivery to both sides of the pulmonary parenchyma need to be performed, and the efficacy of this treatment for disease prevention remains to be determined.

The release of neurotransmitters at synapses belongs to the most important processes in the central nervous system. In the last decades much has been learned about the molecular mechanisms which form the basis for this fundamental process. Highly regulated exocytosis, based on the SNARE (soluble N-ethylmaleimide-sensitive attachment protein receptor) complex and its regulatory molecules is the signature specialization of the nervous system and is shared by neurons and neuroendocrine cells. Cells of the immune system use a similar mechanism to release cytotoxic materials from secretory granules at contacts with virally or bacterially infected cells or cancer cells, in order to remove these threats. These contact zones have been termed immunological synapses in reference to the highly specific targeted exocytosis of effector molecules. Recent findings indicate that mutations in SNARE or SNARE-interacting proteins are the basis of a number of devastating immunological diseases. While SNARE complexes are ubiquitous and mediate a wide variety of membrane fusion events it is surprising that in many cases the SNARE proteins involved in immunological synapses are the same molecules which mediate regulated exocytosis of transmitters and hormones in neurons and neuroendocrine cells. These similarities raise the possibility that results obtained at immunological synapses may be applicable, in particular in the area of presynaptic function, to neuronal synapses. Since immunological synapses (IS) are assembled and disassembled in about a half an hour, the use of immune cells isolated from human blood allows not only the study of the molecular mechanisms of synaptic transmission in human cells, but is particularly suited to the examination of the assembly and disassembly of these “synapses” via live imaging. In this overview we discuss areas of similarity between synapses of the nervous and immune systems and in the process will refer to results of our experiments of the last few years.

Zusammenfassung Die Freisetzung von Neurotransmittern an Synapsen gehört zu den wichtigsten Mechanismen im zentralen Nervensystem. In den zurückliegenden Jahrzehnten konnten viele Erkenntnisse über die molekularen Mechanismen, die diesem Prozess zugrunde liegen, gesammelt werden. Die hochregulierte Exozytose, die auf dem SNARE-Komplex („soluble N-ethylmaleimide-sensitive factor attachment protein receptor“) und seinen regulatorischen Molekülen basiert, ist das Merkmal des Nervensystems sowohl in Neuronen als auch in neuroendokrinen Zellen. Zellen des Immunsystems benutzen einen ähnlichen Mechanismus, um zytotoxische Substanzen aus sekretorischen Granulen freizusetzen. Diese Sekretion findet an Kontaktzonen mit Zelle statt, die mit Viren oder Bakterien infiziert sind sowie Krebszellen, um diese Bedrohung zu beseitigen. Diese Kontaktzonen werden als immunologische Synapsen bezeichnet im Hinblick auf die hochspezifische, zielgerichtete Exozytose von Effektormolekülen. Aktuelle Studien haben gezeigt, dass Mutationen in den SNARE oder SNARE-interagierenden Proteinen die Grundlage für zahlreiche schwerwiegende immunologische Erkrankungen sind. Obwohl SNARE-Komplexe ubiquitär vorkommen und eine große Vielfalt an Fusionsereignissen an der Membran vermitteln, ist es überraschend, dass in vielen Fällen die gleichen SNARE – Proteine an der immunologischen Synapse beteiligt sind, die die Regulation der Exozytose von Transmittern und Homonen in Neuronen und neuroendokrinen Zellen vermitteln. Diese Ähnlichkeiten zeigen die Möglichkeit auf, dass Erkenntnisse, die von immunologischen Synapsen erhalten wurden, auch auf neuronale Synapsen zutreffen, insbesondere im Bereich der präsynaptischen Funktion. Da immunologische Synapsen (IS) innerhalb von etwa 30 Minuten gebildet und wieder abgebaut werden, ermöglicht die Verwendung von Immunzellen, die aus humanem Blut gewonnen wurden, nicht nur die Untersuchung der molekularen Mechanismen der synaptischen Transmission in menschlichen Zelle, sondern auch Untersuchungen der Bildung und des Abbaus dieser „Synapsen“ mittels bildgebender Verfahren. In diesem Übersichtartikel vergleichen wir die Ähnlichkeit der Synapsen des Nerven- und Immunsystems und gehen dabei auf unsere Erkenntnisse der Arbeiten der letzten Jahre ein.

The development of economic indexing systems for ranking cultivars of forage species requires new knowledge of factors that may influence the scaling of agronomic data collected in controll...

In acutely ill patients, particularly in intensive care units or in mixed infections, time to a microbe-specific diagnosis is critical to a successful outcome of therapy. We report the application of evolving technologies involving mass spectrometry to diagnose and monitor a patient’s course. As proof of this concept, we studied five patients and used two rat models of mono-infection and coinfection. We report the noninvasive combined monitoring of Aspergillus fumigatus and Pseudomonas aeruginosa infection. The invasive coinfection was detected by monitoring the fungal triacetylfusarinine C and ferricrocin siderophore levels and the bacterial metabolites pyoverdin E, pyochelin, and 2-heptyl-4-quinolone, studied in the urine, endotracheal aspirate, or breath condensate. The coinfection was monitored by mass spectrometry followed by isotopic data filtering. In the rat infection model, detection indicated 100-fold more siderophores in urine compared to sera, indicating the diagnostic potential of urine sampling. The tools utilized in our studies can now be examined in large clinical series, where we could expect the accuracy and speed of diagnosis to be competitive with conventional methods and provide advantages in unraveling the complexities of mixed infections.

BACKGROUND Although it is known that oral antihistamine-pseudoephedrine combination tablets have a faster onset than intranasal corticosteroid sprays in the treatment of allergic rhinitis after the first dose, the magnitude of change has not been measured in a comparative manner. Furthermore, the sensation of sprayed liquid in the nose may lead patients to mistakenly believe that intranasal steroid sprays work instantly. OBJECTIVE To evaluate, numerically, nasal airflow changes provided by a single dose of loratadine-pseudoephedrine tablet (LP) and fluticasone propionate nasal spray (FP) in participants experiencing allergic rhinitis symptoms, including nasal congestion. METHODS This single-center, double-blinded, placebo-controlled, crossover study evaluated objective nasal airflow changes in patients with a documented sensitivity to ragweed pollen. Participants were randomized to receive 1 of 4 treatment sequences, and their peak nasal inspiratory flow (PNIF) was measured in a span of 4 hours after pollen exposure in an environmental exposure unit. RESULTS Average change in PNIF was 31% with LP in the course of the study, significantly greater than with placebo and FP (12% and 15%, respectively; P < .001). Nevertheless, FP did not produce a significant change compared with its placebo. At hour one post-dose, LP had a clinically significant 31% increase in PNIF, whereas FP only yielded an 8.6% increase (P < .001). Measurable nasal airflow improvements are associated with the opening of nasal passages, allowing congested patients to breathe more freely. CONCLUSION A single dose of LP quickly and significantly (P < .001) improved nasal airflow after ragweed pollen challenge in an environmental exposure unit. Comparatively, FP did not display this same benefit. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT03443843.

A 56-year-old Hispanic man with a history of disseminated coccidioidomycosis was diagnosed with persistent glucocorticoid insufficiency and pseudohyperaldosteronism secondary to posaconazole toxicity. This case was notable for unexpected laboratory findings of both pseudohyperaldosteronism and severe glucocorticoid deficiency due to posaconazole’s mechanism of action on the adrenal steroid synthesis pathway. Transitioning to fluconazole and starting hydrocortisone resolved the hypokalemia but not his glucocorticoid deficiency. This case highlights the importance of recognizing iatrogenic glucocorticoid deficiency with azole antifungal agents and potential long term sequalae.

A virus-free (VF) A. fumigatus isolate has been shown to be resistant in competition with Pseudomonas as compared to the isogenic line infected with Aspergillus fumigatus polymycovirus 1 (AfuPmV-1), and this phenotype was apparently related to alterations in iron metabolism. Here we investigated further the mechanisms underpinning this phenotype. The extracellular siderophore profiles of five isogenic VF and virus-infected (VI) strains were sampled at 24, 31, 48, 54, and 72 h in submerged cultures and quantitatively examined by liquid chromatography and mass spectrometry. Intracellular profiles of conidia and cultures at the stationary growth phase were defined. VF A. fumigatus demonstrated the best fitness represented by the fastest onset of its exponential growth when grown on an iron-limited mineral medium. The exponential phase and transitional production phase of the extracellular triacetylfusarinine C (TafC) were achieved at 24 and 31 h, respectively, contrary to VI strains, which acted more slowly. As a result, the TafC reservoir was consumed sooner in the VF strain. Additionally, the VF strain had lower ferricrocin and higher hydroxyferricrocin content in the pellet during the stationary phase. All of these differences were significant (Kruskal–Wallis, p &lt, 0.01). In our study, the siderophore reservoir of a VF strain was consumed sooner, improving the fitness of the VF strain in competition with P. aeruginosa.

Background: The CMDA/MedSend Healthcare Missions Leadership Summit is an annual gathering of leaders from 75 mission sending agencies who work around the world in healthcare missions. The summit is jointly sponsored by MedSend and the Christian Medical & Dental Associations (CMDA). The purpose of the meeting is to share best practices, network, and examine cutting edge research issues in the field of healthcare missions. Objective: The purpose of the 2015 Launch survey was to learn from medical missionaries how to more effectively recruit and launch new healthcare professionals and their families into long-term missions. The objective of the 2016 Relocating for the Gospel survey was to learn from students and other aspiring healthcare missionaries the obstacles they perceive in their journey into long-term missions. Results were presented at the 2015 and 2016 Healthcare Missions Leadership Summits. Methods: Two multiple-choice surveys were developed to ascertain the positive and negative factors involved in the decision making process to launch into the healthcare mission field. Participants were invited to participate in the surveys via email. Results: The 2015 Launch survey analyzed 177 responses from healthcare missionaries with 63% of respondents still currently serving as long-term missionaries. 37% of the respondents were missionaries previously. Participants included millennials (n=39), Gen Xers (n=54), and baby boomers (n=84). Responses indicated that key positive factors include personal interaction with a long-term worker as well as a supportive agency, leader, or team to join. The 2016 Relocating for the Gospel survey analyzed responses from students and other aspiring missionaries who are millennials (n=79). The strongest overall factor that discouraged missions involvement was student loan debt. Other key obstacles include concerns over being lonely or isolated overseas, as well as difficulty in finding a good fit with a team or organization. Conclusion: The overall findings from the two surveys include four main points. First, effective launch into missions service is more relationally dependent than informationally dependent. Second, launching into missions service is more like a marathon than a sprint. Third, those who are exploring missions want to join a team so they will not feel lonely or isolated. Finally, aspiring missionaries want a placement that fits them well even if that does not match the top priorities of the agency.

Acute and chronic infection with Trypanosoma cruzi affects millions of people. The current therapeutic options are highly toxic and often not effective. Liposomal amphotericin B (LAMB) has been demonstrated previously to have some activity in murine models. In our studies, higher dosages given multiple times were tested for activity against acute or chronic disease, exploring whether intermittent and brief regimens could be effective, as might then prove useful in human, particularly outpatient, therapy. For acute infection, LAMB 25 mg/kg intravenously (i.v.) given one to three times prolonged survival and caused a rapid disappearance of Y strain trypomastigotes from the blood. However, even four or six doses of LAMB 30 mg/kg i.v., did not result in the cure of Y strain infection, with all mice relapsing after being immunosuppressed with cyclophosphamide. Similarly, chronic infection due to the CL strain was found to be unaltered by 1-3 treatments with LAMB 25 mg/kg. All surviving mice had histopathological evidence of infection in one or more tissues and equivalent antibody titers regardless of treatment regimen. Overall, LAMB at doses up to 30 mg/kg i.v. prolonged survival, but these doses were not curative in the regimens studied.

Pseudomonas aeruginosa and Aspergillus fumigatus are major microbes in cystic fibrosis (CF). We reported non-mucoid P. aeruginosa isolates more inhibitory to A. fumigatus than mucoid ones. Another CF P. aeruginosa phenotype, small colony variants (SCVs), is an unknown factor in intermicrobial competition with A. fumigatus. Clinical SCV isolates and reference CF non-mucoid isolate (Pa10, producing normal-sized colonies) were compared. Live cells of P. aeruginosa or filtrates from P. aeruginosa planktonic or biofilm cultures were co-incubated with A. fumigatus growing under conditions allowing biofilm formation or with preformed biofilm. Metabolic activity of A. fumigatus biofilm was then measured. When necessary, assays were done after adjustment for growth differences by adding fresh medium to the planktonic culture filtrate. Pyoverdine determinations were performed spectrophotometrically on the planktonic culture filtrates. In all experimental conditions (live cells and planktonic or biofilm culture filtrates of P. aeruginosa versus A. fumigatus biofilm formation or preformed biofilm), three SCV isolates were less inhibitory than Pa10, two equal or more inhibitory. Adjusting planktonic culture filtrates for growth differences showed SCV inhibition differences variably related to growth or deficient inhibitor production. Studies suggested the principal P. aeruginosa inhibitor to be pyoverdine. SCV isolates appear heterogeneous in their capacity to inhibit A. fumigatus biofilm. SCV isolates can be important in the CF microbiome, because they are capable of intermicrobial inhibition.

Pseudomonas aeruginosa (Pa) and Aspergillus fumigatus (Af) are the major bacterial and fungal pathogens in the airways of cystic fibrosis (CF) patients. This is likely related to their ability to form biofilms. Both microbes have been associated with CF disease progression. The interplay between these two pathogens has been studied under aerobic conditions, though accumulating data indicates that much of the CF airway is hypoxic or anaerobic. We studied the microbial interaction in these latter environments. Pa is an aggressor against Af forming biofilm or as established Af biofilm, whether Pa is cultivated in aerobic, hypoxic, or anaerobic conditions, or tested in aerobic or hypoxic conditions. Pa cells are generally more effective than planktonic or biofilm culture filtrates. Pa growth is less in anaerobic conditions, and filtrates less effective after anaerobic or hypoxic growth, or against hypoxic Af. These, and other comparisons shown, indicate that Pa would be less effective in such environments, as would be the case in a CF mucus plug. These observations would explain why Pa becomes established in CF airways before Af, and why Af may persist during disease progression.

RNA enzymes (ribozymes) have remarkably diverse biological roles despite having limited chemical diversity. Protein enzymes enhance their reactivity through recruitment of cofactors; likewise, the naturally occurring glmS ribozyme uses the glucosamine-6-phosphate (GlcN6P) organic cofactor for phosphodiester bond cleavage. Prior structural and biochemical studies have implicated GlcN6P as the general acid. Here we describe new catalytic roles of GlcN6P through experiments and calculations. Large stereospecific normal thio effects and a lack of metal-ion rescue in the holoribozyme indicate that nucleobases and the cofactor play direct chemical roles and align the active site for self-cleavage. Large stereospecific inverse thio effects in the aporibozyme suggest that the GlcN6P cofactor disrupts an inhibitory interaction of the nucleophile. Strong metal-ion rescue in the aporibozyme reveals that this cofactor also provides electrostatic stabilization. Ribozyme organic cofactors thus perform myriad catalytic roles, thereby allowing RNA to compensate for its limited functional diversity.

Improved performance of the national ewe flock along with the displacement of finishing farms into hill country has increased pressure on spring pasture production. Increasing spring pasture production to cope with this increased demand is considered one of the main areas of improvement needed in hill country farming. A trial was set up in the South Island of New Zealand to determine the response of spring pasture production to two winter grazing intensities across three levels of sward dead material. Total spring pasture production was unaffected by winter grazing intensity. Winter dead material levels significantly affected total spring pasture production. Plots with high dead material levels (>60%) grew 657 kg DM/ha less than plots with low dead material levels (

Artificial rearing is used routinely on dairy sheep farms. Different strategies are used to optimise the growth and development of the young lamb with an economic ration of milk and other feed components. Early weaning, and early introduction to pasture can both reduce these costs, but may also reduce the liveweight gains of the growing lamb. Can differences in pre-weaning feeding strategies be mitigated using specialist pastures such as plantain/red/white clover? Lambs from an experiment investigating the impacts of rearing with and without meal access (n=30/group) and weaned at 12 weeks of age were grazed on either perennial ryegrass-based or plantain/red clover-based pastures in 3 replicates. Liveweight gain, animal health, feed supply and feed quality parameters were recorded over the following 5 weeks. Liveweights of the lambs reared on the different feeding regimens were similar at 12 weeks of age when the grazing study began (25.9 kg). Using plantain/red clover-based pastures provided no advantage to the liveweight gain of lambs reared under different milk and concentrate feeding regimens. Both feed types provided adequate nutrition to ensure growth rates of approximately 180 g/day over the 5 week measurement period in late spring. However, lambs reared under a high concentrate system has consistently lower liveweight gains (160 g/day) over the 5 weeks than those lambs reared with access to pasture (200 g/day; P

Aspergillus fumigatus biofilms consist of a three-dimensional network of cellular hyphae and extracellular matrix. They are involved in infections of immune-compromised individuals, particularly those with cystic fibrosis. These structures are associated with persistence of infection, resistance to host immunity, and antimicrobial resistance. Thorough understanding of structure and function is imperative in the design of therapeutic drugs. Optimization of processing parameters, including aldehyde fixation, heavy metal contrasting, drying techniques and Ionic Liquid treatment, was undertaken for an ultrastructural approach to understand cellular and extracellular biofilm components. Conventional and Variable Pressure Scanning Electron Microscopy were applied to analyze the structure of biofilms attached to plastic and formed at an air-liquid interface.

Background:The optimal first-line treatment of patients (pts) with early rheumatoid arthritis (RA) is yet to be established.Objectives:The primary aim was to assess and compare the proportion of pts who achieved remission with active conventional therapy (ACT) and with three different biologic therapies after 24 wks. Secondary aims were to assess and compare other efficacy measures.Methods:The investigator-initiated NORD-STAR trial (NCT01491815) was conducted in the Nordic countries and Netherlands. In this multicenter, randomized, open-label, blinded-assessor study pts with treatment-naïve, early RA with DAS28>3.2, and positive RF or ACPA, or CRP >10mg/L were randomized 1:1:1:1. Methotrexate (25 mg/week after one month) was combined with: 1) (ACT): oral prednisolone (tapered quickly);or: sulphasalazine, hydroxychloroquine and mandatory intra-articular (IA) glucocorticoid (GC) injections in swollen joints Results:812 pts were randomized. Age was 54.3±14.7 yrs (mean±SD), 31.2% were male, DAS28 5.0±1.1, 74.9% were RF and 81.9% ACPA positive. Fig 1 shows the adjusted CDAI remission rates over time with 95% CI. Table shows crude remission and response rates and absolute differences in adjusted remission and response rates (superiority analysis). Differences in remission and response rates with CZP and TCZ, but not with ABA, remained within the pre-defined non-inferiority margin versus ACT, Fig 2.Figure 1.CDAI remission over time (adj. estimates with 95% CI)Figure 2.Non-inferiority analysis of protocol population. Estimated differences in CDAI remission rates between Arm 1 (active conventional therapy) and Arms 2, 3, and 4 (biologic arms) as reference with 95% confidence intervals, adjusted for gender, ACPA status, country, age, body-mass index and baseline DAS28-CRP. ABA, abatacept; CZP, certolizumab-pegol; MTX, methotrexate; TCZ, tocilizumab.Conclusion:High remission rates were found across all four treatment arms at 24 wks. Higher CDAI remission rate was observed for ABA versus ACT (+9%) and for CZP (+4%), but not for TCZ (-1%). With the predefined 15% margin, ACT was non-inferior to CZP and TCZ, but not to ABA. This underscores the efficacy of active conventional therapy based on MTX combined with glucocorticoids and may guide future treatment strategies for early RA.Table.Primary and key secondary outcomes at 24 weeks (ITT)Active conventional therapy (ACT)Certolizumab+MTXAbatacept+MTXTocilizumab+MTXNo of pts (ITT)200203204188§Crude remission and response ratesCDAI remission42.0%47.8%52.5%41.0%ACR/EULAR Boolean remission34.0%38.4%37.3%31.4%DAS28 remission63.5%68.5%69.6%63.3%SDAI remission41.5%49.8%51.5%42.6%EULAR good response71.5%76.9%79.9%71.3%Difference (95% CI) in rates with Arm 1 as reference (adjusted)CDAI remissionRef4% (-5 to 13%)9% (0.1 to 19%)-1% (-10 to 9%)ACR/EULAR Boolean remissionRef4% (-6 to 13%)5% (-5 to 14%)-4% (-13 to 6%)DAS28 remissionRef3% (-6 to 11%)5% (-4 to 13%)-1% (-10 to 8%)SDAI remissionRef6% (-3 to 18%)9% (-0.3 to 18%)1% (-8 to 11%)EULAR good responseRef4% (-4 to 14%)8% (-2 to 18%)0.4% (-10 to 11%)§17 patients allocated to Tocilizumab did not receive it due to its unavailability and were excluded from ITT.Acknowledgments:Manufacturers provided CZP and ABA.Disclosure of Interests:Merete L. Hetland Grant/research support from: BMS, MSD, AbbVie, Roche, Novartis, Biogen and Pfizer, Consultant of: Eli Lilly, Speakers bureau: Orion Pharma, Biogen, Pfizer, CellTrion, Merck and Samsung Bioepis, Espen A Haavardsholm Grant/research support from: AbbVie, UCB Pharma, Pfizer Inc, MSD Norway, Roche Norway, Consultant of: Pfizer, AbbVie, Janssen-Cilag, Gilead, UCB Pharma, Celgene, Lilly, Paid instructor for: UCB Pharma, Speakers bureau: Pfizer, AbbVie, UCB Pharma, Celgene, Lilly, Roche, MSD, Anna Rudin Consultant of: Astra/Zeneca, Dan Nordström Consultant of: Abbvie, Celgene, Lilly, Novartis, Pfizer, Roche and UCB., Speakers bureau: Abbvie, Celgene, Lilly, Novartis, Pfizer, Roche and UCB., Michael Nurmohamed Grant/research support from: Not related to this research, Consultant of: Not related to this research, Speakers bureau: Not related to this research, Björn Gudbjornsson Speakers bureau: Novartis and Amgen, Jon Lampa Speakers bureau: Pfizer, Janssen, Novartis, Kim Hørslev-Petersen: None declared, Till Uhlig Consultant of: Lilly, Pfizer, Speakers bureau: Grünenthal, Novartis, Gerdur Gröndal: None declared, Mikkel Ǿstergaard Grant/research support from: AbbVie, Bristol-Myers Squibb, Celgene, Merck, and Novartis, Consultant of: AbbVie, Bristol-Myers Squibb, Boehringer Ingelheim, Celgene, Eli Lilly, Hospira, Janssen, Merck, Novartis, Novo Nordisk, Orion, Pfizer, Regeneron, Roche, Sandoz, Sanofi, and UCB, Speakers bureau: AbbVie, Bristol-Myers Squibb, Boehringer Ingelheim, Celgene, Eli Lilly, Hospira, Janssen, Merck, Novartis, Novo Nordisk, Orion, Pfizer, Regeneron, Roche, Sandoz, Sanofi, and UCB, Marte Heiberg: None declared, Jos Twisk: None declared, Simon Krabbe: None declared, Kristina Lend: None declared, Inge Olsen: None declared, Joakim Lindqvist: None declared, Anna-Karin H Ekwall Consultant of: AbbVie, Pfizer, Kathrine L. Grøn Grant/research support from: BMS, Meliha C Kapetanovic: None declared, Francesca Faustini: None declared, Riitta Tuompo: None declared, Tove Lorenzen: None declared, Giovanni Cagnotto: None declared, Eva Baecklund: None declared, Oliver Hendricks Grant/research support from: Pfizer, MSD, Daisy Vedder: None declared, Tuulikki Sokka-Isler: None declared, Tomas Husmark: None declared, Maud-Kristine A Ljosa: None declared, Eli Brodin: None declared, Torkell Ellingsen: None declared, Annika Soderbergh: None declared, Milad Rizk Speakers bureau: AbbVie, Åsa Reckner: None declared, Per Larsson: None declared, Line Uhrenholt Speakers bureau: Abbvie, Eli Lilly and Novartis (not related to the submitted work), Søren Andreas Just: None declared, David Stevens: None declared, Trine Bay Laurberg Consultant of: UCB Pharma (Advisory Board), Gunnstein Bakland Consultant of: Novartis, UCB, Ronald van Vollenhoven Grant/research support from: BMS, GSK, Lilly, UCB, Pfizer, Roche, Consultant of: AbbVie, AstraZeneca, Biogen, Biotest, Celgene, Gilead, Janssen, Pfizer, Servier, UCB, Speakers bureau: AbbVie, Pfizer, UCB

This work was supported by funding from the UK Natural Environment Research Council's iSTAR Program through grants NE/J005703, NE/J005649/1 and NE/J005770/1. The Amundsen Sea is a key region of Antarctica where ocean, atmosphere, sea ice and ice sheet interact. For much of Antarctica, the relatively warm ocean water in the open Southern Ocean (a few degrees above freezing) is unable to reach the continental shelf in large volumes under current climate conditions. In the Amundsen Sea, however, warm water penetrates onto the continental shelf and provides heat that can melt the underside of the floating ice shelves. Here we discuss how the role of the ocean has come under increased scrutiny in recent years, because ocean heat fluxes have been implicated in the thinning of the ice shelves. We present observations from the Amundsen Sea in 2014 and discuss their implications, highlighting aspects where our understanding is still incomplete. Publisher PDF

Diagnosing coccidioidal meningitis (CM) can be problematic owing to its infrequency and/or a delay in the positivity of a cerebrospinal fluid (CSF) culture or CSF antibody, particularly if the primary coccidioidal infection is unrecognized. We tested 37 CSF specimens, 26 from patients with confirmed CM and 11 from patients with suspected microbial meningitis without fungal diagnosis, for (1,3)-beta-glucan (BG). BG in CM CSF specimens ranged from 18 to 3,300 pg/ml and in controls ranged from 103 pg/ml were CM. The one false-negative specimen was from a patient with a pseudosyrinx, without inflammatory evidence of meningitis activity. Serial samples from some patients were positive at ≤8 years, indicating no loss of positivity with chronicity. Samples stored frozen since 2000 included those with 2 of the 3 highest values, indicating that fresh samples not required. A previous study indicated serum sensitivities of 53% in acute, 50% in resolved, and 83% in disseminated and meningeal coccidioidomycosis. Three studies of other fungal meningitides ranged from 86 to 1,524 pg/ml CSF, with 37 controls of

The objective of this experiment was to determine appropriate methods for estimating fecal output, digestibility, and intake in red deer (Cervus elaphus). Dry matter intake (DMI), digestibility, and fecal output were estimated using the dual-marker (titanium dioxide; TiO2 and indigestible acid detergent fiber) technique, double n-alkane ratio technique (ALK) and the pulse dose (Yttrium; Y) technique to determine a suitable method to estimate DMI, fecal output, and digestibility measurements. Four male and four female deer were stratified by sex and randomly assigned either fresh-cut perennial ryegrass (Lolium perenne L.) or fresh-cut plantain (Plantago lanceolata) ad libitum in a cross-over design experiment. Actual DMI (mean ± SD: 1.5 ± 0.36 kg DM/d), digestibility (0.70 ± 0.06), and fecal output (0.45 ± 0.1 kg DM/d) were measured daily over the collection periods, and the average of each period was used for methods’ comparison. The ALK method adequately estimated digestibility and fecal output of plantain; however, overestimated digestibility (P < 0.05) and DMI of ryegrass, so that there was no statistical agreement (P > 0.10) in DMI when diets were pooled. The overestimated DMI of the ryegrass diet led to ALK predicting greater intake when deer consumed ryegrass than plantain, which was the opposite of actual measurements. The pulse dosed Y overestimated (P < 0.05) fecal output and consequently DMI for both plantain and ryegrass, however, indicated similar trends to actual values. The dual-marker technique using TiO2 was able to detect the statistical differences between plantain and ryegrass as the actual measurements, had moderate to strong precision (r = 0.50 to 0.66) and statistical agreement (P < 0.05) with the pooled diet data. Therefore, the dual-marker technique provided the best alternative estimation method to actual measurements of forage DMI of grazing red deer.

Supplementary data are available at MMYCOL online. Aspergillosis is a respiratory fungal disease of importance in captive marine birds. The aim of this study was to describe the occurrence of aspergillosis in Thalassarche melanophris during rehabilitation events and to identify the etiological agent. All the albatrosses that were received for rehabilitation and died within a 2-year period were included in the study. The proportionate mortality rate caused by aspergillosis was 21.4% (3/14). One of the etiological agents was Aspergillus flavus/oryzae lineage, and the other was A. fumigatus sensu stricto. Our study suggests that aspergillosis can act as a limiting factor in the rehabilitation of albatrosses. This study was supported by the International Cooperation Program financed by Brazilian Federal Agency for Support and Evaluation of Graduate Education within the Ministry of Education of Brazil (CAPES), under the Capes-Print Program and PDSE Program - Finance Code 001. info:eu-repo/semantics/publishedVersion

Key Points Question Are childhood behavioral problems associated with self-reported insomnia symptoms in adulthood? Findings In this population-based cohort study, there was a significant association between childhood behavioral problems measured using the Rutter Behavioral Scale at 5, 10, and 16 years of age and self-reported insomnia symptoms at 42 years of age. Meaning There was an increased risk of self-reported insomnia symptoms at 42 years of age among those who had moderate and severe behavioral problems during childhood, compared with those without behavioral problems during childhood., Importance Life-course determinants of insomnia, particularly the long-term association of childhood behavioral problems with insomnia later in life, are unknown. As childhood behaviors are measurable and potentially modifiable, understanding their associations with insomnia symptoms may provide novel insights into early intervention strategies to reduce the burden. Objective To investigate the association between behavioral problems at 5, 10, and 16 years of age and self-reported insomnia symptoms at 42 years of age. Design, Setting, and Participants This cohort study used data from the United Kingdom 1970 Birth Cohort Study, an ongoing large-scale follow-up study. Participants were followed up from birth (1970) to age 42 years (2012). Missing data were imputed via multiple imputation. Statistical analysis was performed from February 1 to July 15, 2019. Exposures Behavior measured at 5, 10, and 16 years of age using the Rutter Behavioral Scale (RBS). Children’s behavior was classified as normal (≤80th percentile), moderate behavioral problems (>80th to ≤95th percentile), and severe behavioral problems (>95th percentile) based on their RBS score. Main Outcomes and Measures Self-reported difficulties initiating or maintaining sleep (DIMS) were collected using a self-administered questionnaire at 42 years of age. Log-binomial logistic regression, adjusted for several potential confounders, was used to estimate the association of childhood behavioral problems with insomnia symptoms in adulthood. Sensitivity analyses were conducted to check robustness of the findings. Results Participants were followed up from a baseline age of 5 years (n = 8050; 3854 boys and 4196 girls), 10 years (n = 9090; 4365 boys and 4725 girls), or 16 years (n = 7653; 3575 boys and 4078 girls) until age 42 years. There was a 39% higher risk of DIMS (odds ratio [OR], 1.39; 95% CI, 1.04-1.84; P = .06 for trend) for participants with severe behavioral problems at 5 years of age compared with those with a normal RBS score. The odds of DIMS plus not feeling rested on waking (DIMS plus) in participants with severe behavioral problems at 5 years of age were 29% higher (odds ratio, 1.29; 95% CI, 0.97-1.70; P = .14 for trend) than participants with a normal RBS score, although this result was not statistically significant. Moderate and severe behavioral problems at 16 years of age were positively associated with DIMS and DIMS plus (moderate: OR, 1.28; 95% CI, 1.07-1.52; severe: OR, 1.67; 95% CI, 1.22-2.30; P, This cohort study investigates the association between behavioral problems at 5, 10, and 16 years of age and self-reported insomnia symptoms at 42 years of age.

Trypanosoma cruzi is the etiologic agent of Chagas disease (CD), which can result in severe cardiomyopathy. Trypanosoma cruzi is endemic to the Americas, and of particular importance in Latin America. In the United States and other non-endemic countries, rising case numbers have also been observed. The currently used drugs are benznidazole (BNZ) and nifurtimox, which have limited efficacy during chronic infection. We repurposed itraconazole (ICZ), originally an antifungal, in combination with amiodarone (AMD), an antiarrhythmic, with the goal of interfering with T. cruzi infection. Human pluripotent stem cells (hiPSCs) were differentiated into cardiomyocytes (hiPSC-CMs). Vero cells or hiPSC-CMs were infected with T. cruzi trypomastigotes of the II or I strain in the presence of ICZ and/or AMD. After 48 hours, cells were Giemsa stained, and infection and multiplication were evaluated microscopically. Trypanosoma cruzi infection and multiplication were evalutated also by electron microscopy. BNZ was used as a reference compound. Cell metabolism in the presence of test substances was assessed. Itraconazole and AMD showed strain- and dose-dependent interference with T. cruzi infection and multiplication in Vero cells or hiPSC-CMs. Combinations of ICZ and AMD were more effective against T. cruzi than the single substances, or BNZ, without affecting host cell metabolism, and better preserving host cell integrity during infection. Our in vitro data in hiPSC-CMs suggest that a combination of ICZ and AMD might serve as a treatment option for CD in patients, but that different responses due to T. cruzi strain differences have to be taken into account.

A novel immunotherapy targeting the Mucorales unique CoH3 invasin is highly protective against murine mucormycosis., Mucorales are fungal pathogens that cause mucormycosis, a lethal angioinvasive disease. Previously, we demonstrated that Rhizopus, the most common cause of mucormycosis, invades endothelial cells by binding of its CotH proteins to the host receptor GRP78. Loss of CotH3 renders the fungus noninvasive and attenuates Rhizopus virulence in mice. Here, we demonstrate that polyclonal antibodies raised against peptides of CotH3 protected diabetic ketoacidotic (DKA) and neutropenic mice from mucormycosis compared to mice treated with control preimmune serum. Passive immunization with anti-CotH3 antibodies enhanced neutrophil inlfux and triggered Fc receptor-mediated enhanced opsonophagocytosis killing of Rhizopus delemar. Monoclonal antibodies raised against the CotH3 peptide also protected immunosuppressed mice from mucormycosis caused by R. delemar and other Mucorales and acted synergistically with antifungal drugs in protecting DKA mice from R. delemar infection. These data identify anti-CotH3 antibodies as a promising adjunctive immunotherapeutic option against a deadly disease that often poses a therapeutic challenge.

Polymeric prodrugs have become an increasingly popular strategy for improving the pharmacokinetic properties of active pharmaceutical ingredients (API). Therefore, identifying a robust method for quantification of the API in these prodrug products is a key part of the drug development process. Current drug quantification methods include hydrolysis followed by reversed phase high-performance liquid chromatography (RP-HPLC), size exclusion chromatography (SEC)-based molecular weight determination, and mass spectrometry. These methods tend to be time-consuming and often require challenging method development. Here, we present a comparative study highlighting the automated elemental analyzer as a facile approach to drug quantification in this up-and-coming class of therapeutics. A polymeric prodrug using poly(L-lysine succinylated) (PLS) and the drug lamivudine (LAM) was prepared and analyzed using the elemental analyzer in comparison to the traditional approaches of hydrolysis followed by RP-HPLC and SEC using multi-angle light scattering (MALS) detection. The elemental analysis approach showed excellent agreement with the conventional methods but proved much less laborious, highlighting this as a rapid and sensitive analytical method for the quantitative determination of drug loading in polymeric prodrug products.

International audience; Pseudomonas aeruginosa and Aspergillus fumigatus are pathogens frequently co-inhabiting immunocompromised patient airways, particularly in people with cystic fibrosis. Both microbes depend on the availability of iron, and compete for iron in their microenvironment. We showed previously that the P. aeruginosa siderophore pyoverdine is the main instrument in battling A. fumigatus biofilms, by iron chelation and denial of iron to the fungus. Here we show that A. fumigatus siderophores defend against anti-fungal P. aeruginosa effects. P. aeruginosa supernatants produced in the presence of wildtype A. fumigatus planktonic supernatants (Afsup) showed less activity against A. fumigatus biofilms than P. aeruginosa supernatants without Afsup, despite higher production of pyoverdine by P. aeruginosa. Supernatants of A. fumigatus cultures lacking the sidA gene (AfΔsidA), unable to produce hydroxamate siderophores, were less capable of protecting A. fumigatus biofilms from P. aeruginosa supernatants and pyoverdine. AfΔsidA biofilm was more sensitive towards inhibitory effects of pyoverdine, the iron chelator deferiprone (DFP), or amphothericin B than wildtype A. fumigatus biofilm. Supplementation of sidA-deficient A. fumigatus biofilm with A. fumigatus siderophores restored resistance to pyoverdine. The A. fumigatus siderophore production inhibitor celastrol sensitized wildtype A. fumigatus biofilms towards the anti-fungal activity of DFP. In conclusion, A. fumigatus hydroxamate siderophores play a pivotal role in A. fumigatus competition for iron against P. aeruginosa.