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Impacts of invasive species are often difficult to quantify, meaning that many invaders are prioritised for management without robust, contextual evidence of impact. Most impact studies for invasive plants compare heavily invaded with non-invaded sites, revealing little about abundance–impact relationships. We examined effects of increasing cover and volume of the non-native herbaceous groundcover Tradescantia fluminensis on a temperate rainforest community of southern Australia. We hypothesised that there would be critical thresholds in T. fluminensis abundance, below which the native plant community would not be significantly impacted, but above which the community’s condition would degrade markedly. We modelled the abundance–impact relationship from 83 plots that varied in T. fluminensis abundance and landscape context and found the responses of almost all native plant indicators to invasion were non-linear. Native species richness, abundance and diversity exhibited negative exponential relationships with increasing T. fluminensis volume, but negative threshold relationships with increasing T. fluminensis cover. In the latter case, all metrics were relatively stable until cover reached between 20 and 30%, after which each decreased linearly, with a 50% decline occurring at 75–80% invader cover. Few growth forms (notably shrubs and climbers) exhibited such thresholds, with most exhibiting negative exponential relationships. Tradescantia fluminensis biomass increased dramatically at > 80% cover, with few native species able to persist at such high levels of invasion. Landscape context had almost no influence on native communities, or the abundance–impact relationships between T. fluminensis and the plant community metrics. Our results suggest that the diversity of native rainforest community can be maintained where T. fluminensis is present at moderate-to-low cover levels.

Purpose We sought to determine the rate of return to play (RTP) in baseball players following arthroscopic elbow debridement for the management of the symptomatic elbow. Methods A retrospective case series with prospectively collected data via questionnaire was conducted on all baseball players who underwent an arthroscopic elbow debridement, from July, 15, 2004 to November 1, 2017. A postoperative questionnaire was released at an average 7.25 year follow-up. Data collected included age, gender, laterality, preoperative diagnosis, range of motion, duration and characterization of symptoms, visual analog scale (VAS) pain score, complications, level of play, and RTP. Results Follow-up data were available on 18 baseball players. Average age was 19.7 years (range 16-24). Seventeen were pitchers, and 1 was a catcher. Level of play included 12 collegiate athletes, 2 high school athletes, 2 recreational athletes, 1 minor league athletes, and 1 major league athlete. Rate of RTP was 61% (11/18) with 6 returning to a greater level and 5 to an equal level. The length of time to RTP following surgery was most commonly within 6-8 months (44.4%, 8/18). Mean VAS pain score improved from 6.9 to 0.75 (P =

The histone deacetylases (HDACs) are a family of enzymes that catalyse lysine deacetylation of both histone and non-histone proteins. Here, we review, summarize, and provide perspectives on the literature regarding one such HDAC, HDAC1, in endothelial biology. In the endothelium, HDAC1 mediates the effects of external and environmental stimuli by regulating major endothelial functions such as angiogenesis, inflammatory signalling, redox homeostasis, and nitric oxide signalling. Angiogenesis is most often, but not exclusively, repressed by endothelial HDAC1. The regulation of inflammatory signalling is more complex as HDAC1 promotes or suppresses inflammatory signalling depending upon the environmental stimuli. HDAC1 is protective in models of atherosclerosis where loss of HDAC1 results in increased cytokine and cell adhesion molecule (CAM) abundance. In other models, HDAC1 promotes inflammation by increasing CAMs and repressing claudin-5 expression. Consistently, from many investigations, HDAC1 decreases antioxidant enzyme expression and nitric oxide production in the endothelium. HDAC1 decreases antioxidant enzyme expression through the deacetylation of histones and transcription factors, and also regulates nitric oxide production through regulating both the expression and activity of nitric oxide synthase 3. The HDAC1-dependent regulation of endothelial function through the deacetylation of both histone and non-histone proteins ultimately impacts whole animal physiology and health.

Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) has become a global pandemic. Case identification is currently made by real-time polymerase chain reaction (PCR) during the acute phase and largely restricted to healthcare laboratories. Serological assays are emerging but independent validation is urgently required to assess their utility. We evaluated five different point-of-care (POC) SARS-CoV-2 antibody test kits against PCR, finding concordance across the assays (n = 15). We subsequently tested 200 patients using the OrientGene COVID-19 IgG/IgM Rapid Test Cassette and find a sensitivity of 74% in the early infection period (day 5–9 post symptom onset), with 100% sensitivity not seen until day 13, demonstrating inferiority to PCR testing in the infectious period. Negative rate was 96%, but in validating the serological tests uncovered potential false-negatives from PCR testing late-presenting cases. A positive predictive value (PPV) of 37% in the general population precludes any use for general screening. Where a case definition is applied however, the PPV is substantially improved (95.4%), supporting use of serology testing in carefully targeted, high-risk populations. Larger studies in specific patient cohorts, including those with mild infection are urgently required to inform on the applicability of POC serological assays to help control the spread of SARS-CoV-2 and improve case finding of patients that may experience late complications.

Although there is a commonly held belief within the baseball community that delivery from the stretch confers more stress at the elbow and shoulder joints than delivery from the windup, there remains little evidence in the literature investigating this hypothesis. This study aimed to help address this gap in the literature by studying both intra-pitcher kinematic sequence variability, and intra-pitcher joint torque variability when throwing from the windup vs. the stretch. We hypothesized that 1) each pitchers’ kinematic sequence would remain similar whether throwing from the windup or stretch, and 2) Kinematic sequence would influence peak arm torque more than delivery method. This cross-sectional 3D biomechanical study included 88 pitches thrown by ten (6 collegiate, 4 high school) pitchers with a mean age of 17.60 ± 2.63 years. Pitch velocity, throwing shoulder/elbow torques and the kinematic sequence of each pitch utilizing segmental peak angular velocities were captured. No statistically significant differences in ball velocity (p = 0.17), peak shoulder external rotation torque (p = 0.80), shoulder extension torque (p = 0.97), or elbow valgus torque (p = 0.83) were found between delivery approaches. Three primary kinematic sequences were identified. Shoulder external rotation torque [F(53,2) = 10.992, ɳ2 = .293, p < 0.00], shoulder extension torque [F(53,2) = 15.517, ɳ2 = .369, p < 0.00] and elbow valgus torque [F(53,2) = 9.994, ɳ2 = .274, p < 0.00] did vary significantly across these three kinematic sequence patterns. Our data suggest that the kinematic sequence influences shoulder and elbow torque more than the delivery approach. Instructing ideal kinematic sequence may be more influential for injury avoidance than delivery method.

Multitarget directed ligands (MTDLs) are emerging as promising treatment options for Alzheimer’s disease (AD). Coumarin derivatives serve as a good starting point for designing MTDLs due to their inherent inhibition of monoamine oxidase (MAO) and cholinesterase enzymes, which are complicit in AD’s complex pathophysiology. A preliminary series of 3,7-substituted coumarin derivatives were synthesised and evaluated for enzyme inhibitory activity, cytotoxicity as well as neuroprotective ability. The results indicated that the compounds are weak cholinesterase inhibitors with five compounds demonstrating relatively potent inhibition and selectivity towards MAO-B with IC50 values between 0.014 and 0.498 hx00B5;µM. Significant neuroprotective effects towards MPP+-compromised SH-SY5Y neuroblastoma cells were also observed, with no inherent cytotoxicity at 10 µM for all compounds. The overall results demonstrated that substitution of the phenylethyloxy moiety at the 7-position imparted superior general activity to the derivatives, with the propargylamine substitution at the 3-position, in particular, displaying the best MAO-B selectivity and neuroprotection.

Background Paget-Schroetter syndrome (PSS) is a rare condition of axillosubclavian vein thrombosis often seen in athletes with a history of repetitive external rotation and abduction of the shoulder. The purpose of this review was to analyze the literature and characterize PSS in the athletic population, including risk of PSS by sport. We also provide a comprehensive review of PSS to inform clinicians on the pathophysiology, detection, and management of the condition. Methods Four databases were reviewed to identify cases of PSS occurring in athletes. Data on patient demographics, reported sport, diagnosis, treatment, management, return to sport, and complications were extracted and analyzed by 2 independent reviewers. Results Of the 123 cases of PSS identified, baseball and weight lifting had the highest incidence (26.8% and 19%, respectively), followed by swimming, football, and basketball. The average return to sport was 4.7 months, and 26.7% of subjects reported complications, most commonly pulmonary embolism. Conclusion In athletes presenting with upper extremity pain and swelling with a history of playing baseball or weight lifting, PSS should be higher on a clinicians differential diagnosis list. Swimmers, football, and basketball players are less likely to present with PSS but are still more likely than other types of athletes to develop the condition. Clinician awareness of PSS in athletes is critical to avoid delays in treatment and misdiagnosis, and to allow for a timely return to sport with minimal complications.

Mutations in nucleotide-binding oligomerization domain-containing protein 2 (NOD2) cause Blau syndrome, an inflammatory disorder characterized by uveitis. The antimicrobial functions of Nod2 are well-established, yet the cellular mechanisms by which dysregulated Nod2 causes uveitis remain unknown. Here, we report a non-conventional, T cell-intrinsic function for Nod2 in suppression of Th17 immunity and experimental uveitis. Reconstitution of lymphopenic hosts with Nod2−/− CD4+ T cells or retina-specific autoreactive CD4+ T cells lacking Nod2 reveals a T cell-autonomous, Rip2-independent mechanism for Nod2 in uveitis. In naive animals, Nod2 operates downstream of TCR ligation to suppress activation of memory CD4+ T cells that associate with an autoreactive-like profile involving IL-17 and Ccr7. Interestingly, CD4+ T cells from two Blau syndrome patients show elevated IL-17 and increased CCR7. Our data define Nod2 as a T cell-intrinsic rheostat of Th17 immunity, and open new avenues for T cell-based therapies for Nod2-associated disorders such as Blau syndrome., How mutations in the microbial receptor NOD2 induce Blau syndrome in humans and related uveitis is unclear. Here the authors show, using Nod2-deficient mice and experimental uveitis, that Nod2 negatively regulates T cell activation and transcription of autoimmunity-related genes to suppress Th17 responses and uveitis.

We describe enantioselective syntheses of strychnos and chelidonium alkaloids. In the first case, indole acetic acid esters were established as excellent partner nucleophiles for enantioselective cooperative isothiourea/Pd catalyzed α-alkylation. This provides products containing indole-bearing stereocenters in high yield and with excellent levels of enantioinduction in a manner that is notably independent of the N-substituent. This led to concise syntheses of (-)-akuammicine and (-)-strychnine. In the second case, the poor performance of ortho-substituted cinnamyl electrophiles in the enantioselective cooperative isothiourea/Ir catalyzed α-alkylation was overcome by appropriate substituent choice, leading to enantioselective syntheses of (+)-chelidonine, (+)-norchelidonine, and (+)-chelamine.

Because of the high diversity of enteric viruses in the environment, there is an increasing need for methods enabling the multiple detection of different pathogens. Quantitative, emerging digital polymerase chain reaction (PCR) and isothermal amplification approaches are capable of quantification of multiple targets and hence are suitable for long-term monitoring and source tracking of enteric viruses in the aquatic environment. The combination of culturing with PCR-based detection enables rapid viral risk assessment, especially with host tissues capable of propagation of several viral strains. Viability assays may provide a better understanding on viral survival than PCR-based approaches alone; however, the usefulness of these assays in waste water and environmental water samples should be further investigated. Undoubtedly, emerging sequencing-based technologies provide invaluable data on the ecology and diversity of viruses and along with rapid on-site technologies, for example, biosensors, may be implemented in viral risk assessment in the aquatic environment in the near future.

Background Stress radiography measures medial joint space opening of the elbow, but its value in the management of throwing athletes is unclear. The purpose of this study was to analyze the relationship between medial joint opening (gapping and excess opening) and ulnar collateral ligament (UCL) injury severity on magnetic resonance imaging, as well as to explore factors related to the unexpected finding of a greater opening of the uninjured elbow compared with the injured elbow (negative excess opening) with valgus stress radiography. Methods Medial joint space measurements were independently performed by 2 raters in a clinical series of 74 patients evaluated with standardized valgus stress radiography as part of their clinical workup for throwing-related medial elbow pain. Demographic data were collected by chart review, and UCL injury severity was classified based on available imaging into intact UCLs, partial-thickness tears of the anterior bundle, or full-thickness tears of the anterior bundle. Results Joint gapping was related to UCL injury severity (P = .003), and group-level comparison showed a difference among tear severity groups (P = .050). Excess opening was not significantly related to UCL injury severity (P = .109). A negative excess opening was observed in 22% of patients, but no factors corroborating guarding or a mechanical explanation were significant for a decreased medial joint opening of the injured elbow compared with the uninjured elbow. Conclusions Medial joint gapping was correlated to UCL injury severity in throwing athletes with medial elbow pain and a clinical suggestion of UCL injury, but no association between injury severity and excess opening was observed in this clinical series, which may limit the usefulness of stress radiography in the clinical workup of throwing athletes.

A series of multi-target directed edaravone derivatives bearing N-benzyl pyridinium moieties were designed and synthesised. Edaravone is a potent antioxidant with significant neuroprotective effects and N-benzyl pyridinium has previously exhibited positive results as part of a dual-site binding, peripheral anionic site (PAS) and catalytic anionic site (CAS), acetylcholinesterase (AChE) inhibitor. The designed edaravone-N-benzyl pyridinium hybrid compounds were docked within the AChE active site. The results indicated interactions with conserved amino acids (Trp279 in PAS and Trp84 in CAS), suggesting good dual-site inhibitory activity. Significant in vitro AChE inhibitory activities were observed for selected compounds (IC50: 1.2–4.6 µM) with limited butyrylcholinesterase inhibitory activity (IC50’s >160 µM), indicating excellent selectivity towards AChE (SI: 46 – >278). The compounds also showed considerable antioxidant ability, similar to edaravone. In silico studies indicated that these compounds should cross the blood–brain barrier, making them promising lead molecules in the development of anti-Alzheimer’s agents.

Insulin receptors are internalized by endothelial cells; however, the impact of hyperinsulinemia on this process is not known. Thus, the aim of this study is to determine the role of hyperinsulinemia on insulin receptor function and internalization, as well as the potential impact of protein tyrosine phosphatase 1B (PTP1B). To this end, hippocampal microvessels were isolated from male C57Bl/6J mice on either a control or high-fat diet and assessed for insulin receptor signaling. Cell surface insulin receptors in brain microvascular endothelial cells were labelled with biotin to assess the role hyperinsulinemia plays on receptor internalization in response to stimulation, with and without Claramine treatment, a potent PTP1B antagonist. Our results indicated that insulin receptor levels increased in tandem with insulin receptor dysfunction in the high-fat diet mouse hippocampal microvessels. Hyperinsulinemic cell-receptors demonstrate a shift in splice variation towards decreased IR-A/IR-B ratios and demonstrate a higher membrane-localized proportion. This corresponded with decreased autophosphorylation at sites critical for receptor internalization and signaling, however, Claramine restored signaling and receptor internalization in hyperinsulinemic cells. In conclusion, hyperinsulinemia negatively impacts brain microvascular endothelial cell insulin receptor function and internalization, likely through both alternative splicing and increased negative feedback from PTP1B.

Most patients with rare diseases do not receive a molecular diagnosis and the aetiological variants and causative genes for more than half such disorders remain to be discovered1. Here we used whole-genome sequencing (WGS) in a national health system to streamline diagnosis and to discover unknown aetiological variants in the coding and non-coding regions of the genome. We generated WGS data for 13,037 participants, of whom 9,802 had a rare disease, and provided a genetic diagnosis to 1,138 of the 7,065 extensively phenotyped participants. We identified 95 Mendelian associations between genes and rare diseases, of which 11 have been discovered since 2015 and at least 79 are confirmed to be aetiological. By generating WGS data of UK Biobank participants2, we found that rare alleles can explain the presence of some individuals in the tails of a quantitative trait for red blood cells. Finally, we identified four novel non-coding variants that cause disease through the disruption of transcription of ARPC1B, GATA1, LRBA and MPL. Our study demonstrates a synergy by using WGS for diagnosis and aetiological discovery in routine healthcare.

This article belongs to the Special Issue Molecular Simulations Applications in Biochemistry and Molecular Biology., The solid-state structural analysis and docking studies of three adamantane-linked 1,2,4-triazole derivatives are presented. Crystal structure analyses revealed that compound 2 crystallizes in the triclinic P-1 space group, while compounds 1 and 3 crystallize in the same monoclinic P21/c space group. Since the only difference between them is the para substitution on the aryl group, the electronic nature of these NO2 and halogen groups seems to have no influence over the formation of the solid. However, a probable correlation with the size of the groups is not discarded due to the similar intermolecular disposition between the NO2/Cl substituted molecules. Despite the similarities, CE-B3LYP energy model calculations show that pairwise interaction energies vary between them, and therefore the total packing energy is affected. HOMO-LUMO calculated energies show that the NO2 group influences the reactivity properties characterizing the molecule as soft and with the best disposition to accept electrons. Further, in silico studies predicted that the compounds might be able to inhibit the 11β-HSD1 enzyme, which is implicated in obesity and diabetes. Self- and cross-docking experiments revealed that a number of non-native 11β-HSD1 inhibitors were able to accurately dock within the 11β-HSD1 X-ray structure 4C7J. The molecular docking of the adamantane-linked 1,2,4-triazoles have similar predicted binding affinity scores compared to the 4C7J native ligand 4YQ. However, they were unable to form interactions with key active site residues. Based on these docking results, a series of potentially improved compounds were designed using computer aided drug design tools. The docking results of the new compounds showed similar predicted 11β-HSD1 binding affinity scores as well as interactions to a known potent 11β-HSD1 inhibitor., This research was funded by the Deanship of Scientific Research at Princess Nourah bint Abdulrahman University through the Research Groups Program (Grant No. RGP-1442-0010-4).

Introduction A systematic literature review was undertaken to evaluate real-world use of ceftazidime-avibactam for infections due to aerobic Gram-negative organisms in adults with limited treatment options. Methods Literature searches retrieved peer-reviewed publications and abstracts from major international infectious disease congresses from January 2015 to February 2021. Results were screened using pre-defined criteria to limit the dataset to relevant publications (notable exclusions were paediatric data and outcomes data for bacteria intrinsically resistant to ceftazidime-avibactam). Data for included publications were subjected to qualitative synthesis. Results Seventy-three relevant publications (62 peer-reviewed articles; 10 abstracts) comprising 1926 patients treated with ceftazidime-avibactam (either alone or combined with other antimicrobials) and 1114 comparator/control patients were identified. All patients were hospitalised for serious illness and most had multiple comorbidities. The most common infections were pneumonia, bacteraemia, and skin/soft tissue, urinary tract, or abdominal infections; smaller numbers of patients with meningitis, febrile neutropenia, osteomyelitis, and cystic fibrosis were also included. Carbapenem-resistant or carbapenemase-producing Enterobacterales (CRE; n = 1718) and carbapenem-resistant, multidrug-resistant (MDR), and extensively drug-resistant Pseudomonas aeruginosa (n = 150) were the most common pathogens. Most publications reported positive outcomes for ceftazidime-avibactam treatment (clinical success rates ranged from 45 to 100% and reported 30-day mortality from 0 to 63%), which were statistically superior versus comparators in some studies. ceftazidime-avibactam resistance emergence occurred infrequently and mostly in Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae strains. Conclusion This review provides qualitative evidence of successful use of ceftazidime-avibactam for the treatment of hospitalised patients with CRE and MDR P. aeruginosa infections with limited treatment options. Supplementary Information The online version contains supplementary material available at 10.1007/s40121-021-00507-6.

Grassland ecosystems form 30-40%1 of total land cover and provide essential ecosystem services, including food production, flood mitigation and carbon storage2. Their productivity is closely related to soil microbial communities3, yet the role of viruses within these critical ecosystems is currently undercharacterised4 and in particular, our knowledge of soil RNA viruses is significantly limited5. Here, we applied viromics6 to characterise soil RNA viral communities along an altitudinal productivity gradient of peat, managed grassland and coastal soils. We identified 3,462 viral operational taxonomic units (vOTUs) and assessed their spatial distribution, phylogenetic diversity and potential host ranges. Soil types exhibited showed minimal similarity in viral community composition, but with >10-fold more vOTUs shared between managed grassland soils when compared with peat or coastal soils. Phylogenetic analyses of viral sequences predicted broad host ranges including bacteria, plants, fungi, vertebrates and invertebrates, contrasting with soil DNA viromes which are typically dominated by bacteriophages7. RNA viral communities therefore likely have the ability to influence soil ecosystems across multiple trophic levels. Our study represents an important step towards the characterisation of terrestrial RNA viral communities and the intricate interactions with their hosts, which will provide a more holistic view of the biology of economically and ecologically important terrestrial ecosystems.

Aminopeptidase N (APN/CD13) is a zinc-dependent M1 aminopeptidase that contributes to cancer progression by promoting angiogenesis, metastasis, and tumor invasion. We have previously identified hydroxamic acid-containing analogues that are potent inhibitors of the APN homologue from the malarial parasite Plasmodium falciparum M1 aminopeptidase (PfA-M1). Herein, we describe the rationale that underpins the repurposing of PfA-M1 inhibitors as novel APN inhibitors. A series of novel hydroxamic acid analogues were developed using a structure-based design approach and evaluated their inhibition activities against APN. N-(2-(Hydroxyamino)-2-oxo-1-(3',4',5'-trifluoro-[1,1'-biphenyl]-4-yl)ethyl)-4-(methylsulfonamido)benzamide (6ad) proved to be an extremely potent inhibitor of APN activity in vitro, selective against other zinc-dependent enzymes such as matrix metalloproteases, and possessed limited cytotoxicity against Ad293 cells and favorable physicochemical and metabolic stability properties. The combined results indicate that compound 6ad may be a useful lead for the development of anticancer agents.

Biodiversity monitoring programs are routinely established to quantify changes in biotic communities in response to land management. Surrogacy is implicitly used in many such monitoring programs whereby the measurement of a component of biodiversity is used to infer responses of broader biodiversity. Yet rarely is this surrogacy validated by demonstrating that measured variables and the target variable of interest have matching responses to management treatments. Here we examined the responses of higher-taxon and functional groupings of ants and birds (our surrogate variables) two years after the implementation of experimental livestock grazing treatments, and compared these with the responses of total ant and bird species richness (our target variables) to the same treatments. We found significant and strong correlations between surrogate and target variables, but this did not predict corresponding similar response to treatments. For ants, we found that the genus Monomorium had a negative response to the grazing exclusion treatment, but there was no matching response of species richness, and so no surrogacy was identified. For birds, total species richness had a weak positive response to spring/summer grazing exclusion, and the abundance of honeyeaters (Meliphagidae) showed a similar positive response, suggesting surrogacy. Our study highlights that correlations among variables do not necessarily lead to surrogacy, and indeed that different sub-components of biotic assemblages can respond in ways that contrast with overall species richness. Careful assessment of the matched responses of surrogate and target variables to management can provide a simple and robust way to critically assess biodiversity surrogacy.

BACKGROUND Injury patterns are well-documented for taekwondo competitions prior to the use of an electronic chest protector for scoring tabulation. To see if injury rates and types changed following this rule change that transformed the fighting style in taekwondo, we investigated injuries in collegiate taekwondo competitions in the United States. METHODS Data were collected at eight collegiate taekwondo tournaments from April 2018 to December 2019. All injured athletes seen at the first-aid station were invited to complete a survey that included injury location, type, and mechanism of injury. Injury rates were calculated per 1,000 athlete-exposures (A-Es) and minute-exposures (M-Es). Risk factors were modeled using logistic regression and chi-squared analysis. RESULTS Out of 1,096 athletes, 194 athletes reported 275 acute injuries. We found an injury risk of 17.7/100 athletes (95% CI 15.4, 20.0) and injury rate of 68.9/1,000 A-E (95% CI 60.7, 77.0) which was comparable to previous studies. The most common injuries were contusions to the lower limbs. In contrast to prior reports, men were injured more frequently from delivering a kick and women from receiving a kick. Populations at higher risk for injury included those with low belt rank and middle weight class for women. CONCLUSIONS It appears that the new fighting style did not affect injury rates. Injury locations and types remain similar, but the mechanisms of injury have reversed as men are more injured from attacking and women from defending. There remains a strong need for research to improve protective equipment and safety rules in taekwondo.

Reduced activity of the insulin/IGF signalling network increases health during ageing in multiple species. Diverse and tissue-specific mechanisms drive the health improvement. Here, we performed tissue-specific transcriptional and proteomic profiling of long-lived Drosophila dilp2-3,5 mutants, and identified tissue-specific regulation of >3600 transcripts and >3700 proteins. Most expression changes were regulated post-transcriptionally in the fat body, and only in mutants infected with the endosymbiotic bacteria, Wolbachia pipientis, which increases their lifespan. Bioinformatic analysis identified reduced co-translational ER targeting of secreted and membrane-associated proteins and increased DNA damage/repair response proteins. Accordingly, age-related DNA damage and genome instability were lower in fat body of the mutant, and overexpression of a minichromosome maintenance protein subunit extended lifespan. Proteins involved in carbohydrate metabolism showed altered expression in the mutant intestine, and gut-specific overexpression of a lysosomal mannosidase increased autophagy, gut homeostasis, and lifespan. These processes are candidates for combatting ageing-related decline in other organisms.

Despite advancements in disease management, sickle cell nephropathy, a major contributor to mortality and morbidity in patients, has limited therapeutic options. Previous studies indicate hydroxyurea, a commonly prescribed therapy for sickle cell disease (SCD), can reduce renal injury in SCD but the mechanisms are uncertain. Because SCD is associated with reduced nitric oxide (NO) bioavailability, we hypothesized that hydroxyurea treatment would improve NO bioavailability in the humanized sickle cell mouse. Humanized male 12-wk-old sickle (HbSS) and genetic control (HbAA) mice were treated with hydroxyurea or regular tap water for 2 wk before renal and systemic NO bioavailability as well as renal injury were assessed. Untreated HbSS mice exhibited increased proteinuria, elevated plasma endothelin-1 (ET-1), and reduced urine concentrating ability compared with HbAA mice. Hydroxyurea reduced proteinuria and plasma ET-1 levels in HbSS mice. Untreated HbSS mice had reduced plasma nitrite and elevated plasma arginase concentrations compared with HbAA mice. Hydroxyurea treatment augmented plasma nitrite and attenuated plasma arginase in HbSS mice. Renal vessels isolated from HbSS mice also had elevated nitric oxide synthase 3 (NOS3) and arginase 2 expression compared with untreated HbAA mice. Hydroxyurea treatment did not alter renal vascular NOS3, however, renal vascular arginase 2 expression was significantly reduced. These data support the hypothesis that hydroxyurea treatment augments renal and systemic NO bioavailability by reducing arginase activity as a potential mechanism for the improvement on renal injury seen in SCD mice.

Background: Burn patients are susceptible to healthcare-associated infections. Contaminated surfaces play a role in microbial transmission. This study aimed to quantify the degree of contamination of burns theatre fomites during routine clinical use. Methods: The Patslide Patient Transfer Board (PAT slide) and operating table were investigated using two methods&mdash, bacterial swabs to culture viable organisms and adenosine triphosphate (ATP) swabs to measure biological material. Both items were sampled four times a day: before the first case, immediately after a case, immediately before the next case after cleaning and after the terminal clean. Results: Among 82 bacterial samples, four organisms were isolated, including Staphylococcus aureus, Enterobacter cloacae (E. cloacae) x2 and Pseudomonas aeruginosa (P. aeruginosa), all from the PAT slide. The E. cloacae persisted after cleaning. In 9/82 swabs, the ATP count was &gt, 10 relative light units (RLU). In all cases where an organism was identified, the ATP count was &gt, 10 RLU. Hence the sensitivity and specificity of ATP &gt, 10 RLU in detecting an organism were 100% and 94% respectively. Conclusions: Within burns theatres, there are instances of bacterial contamination on surfaces that persist despite cleaning. ATP luminometers as a point-of-care device may have a role in determining the cleanliness of surfaces, potentially minimizing onwards-bacterial transmission.

Cytomegaloviruses (CMVs) are highly adapted to their host species resulting in strict species specificity. Hence, in vivo examination of all aspects of CMV biology employs animal models using host-specific CMVs. Infection of rhesus macaques (RM) with rhesus CMV (RhCMV) has been established as a representative model for infection of humans with HCMV due to the close evolutionary relationships of both host and virus. However, the only available RhCMV clone that permits genetic modifications is based on the 68–1 strain which has been passaged in fibroblasts for decades resulting in multiple genomic changes due to tissue culture adaptations. As a result, 68–1 displays reduced viremia in RhCMV-naïve animals and limited shedding compared to non-clonal, low passage isolates. To overcome this limitation, we used sequence information from primary RhCMV isolates to construct a full-length (FL) RhCMV by repairing all mutations affecting open reading frames (ORFs) in the 68–1 bacterial artificial chromosome (BAC). Inoculation of adult, immunocompetent, RhCMV-naïve RM with the reconstituted virus resulted in significant viremia in the blood similar to primary isolates of RhCMV and furthermore led to high viral genome copy numbers in many tissues at day 14 post infection. In contrast, viral dissemination was greatly reduced upon deletion of genes also lacking in 68–1. Transcriptome analysis of infected tissues further revealed that chemokine-like genes deleted in 68–1 are among the most highly expressed viral transcripts both in vitro and in vivo consistent with an important immunomodulatory function of the respective proteins. We conclude that FL-RhCMV displays in vitro and in vivo characteristics of a wildtype virus while being amenable to genetic modifications through BAC recombineering techniques., Author summary Human cytomegalovirus (HCMV) infections are generally asymptomatic in healthy immunocompetent individuals, but HCMV can cause serious disease after congenital infection and in individuals with immunocompromised immune systems. Since HCMV is highly species specific and cannot productively infect immunocompetent laboratory animals, experimental infection of rhesus macaques (RM) with rhesus CMV (RhCMV) has been established as a closely related animal model for HCMV. By employing the unique ability of CMV to elicit robust and lasting cellular immunity, this model has also been instrumental in developing novel CMV-based vaccines against chronic and recurring infections with pathogens such as the human immunodeficiency virus (HIV) and Mycobacterium tuberculosis (Mtb). However, most of this work was conducted with derivatives of the 68–1 strain of RhCMV which has acquired multiple genomic alterations in tissue culture. To model pathogenesis and immunology of clinical HCMV isolates we generated a full-length (FL) RhCMV clone representative of low passage isolates. Infection of RhCMV-naïve RM with FL-RhCMV demonstrated viremia and tissue dissemination that was comparable to that of non-clonal low passage isolates. We further demonstrate that FL-RhCMV is strongly attenuated upon deletion of gene regions absent in 68–1 thus demonstrating the usefulness of FL-RhCMV to study RhCMV pathogenesis.

Simian immunodeficiency virus (SIV) insert-expressing, 68-1 Rhesus Cytomegalovirus (RhCMV/SIV) vectors elicit major histocompatibility complex (MHC)-E- and -II-restricted, SIV-specific CD8+ T cell responses, but the basis of these unconventional responses and their contribution to demonstrated vaccine efficacy against SIV challenge in the rhesus monkeys (RMs) has not been characterized. We demonstrate that these unconventional responses resulted from a chance genetic rearrangement in 68-1 RhCMV that abrogated the function of eight distinct immunomodulatory gene products encoded in two RhCMV genomic regions (Rh157.5/.4 and Rh158-161). Differential repair of these genes with either RhCMV-derived or orthologous human CMV (HCMV)-derived sequences (UL128/130; UL146/147) leads to either of two distinct CD8+ T cell response types – MHC-Ia-restricted-only, or a mix of MHC-II- and MHC-Ia-restricted CD8+ T cells. Despite response magnitude and functional differentiation being similar to RhCMV 68-1, neither alternative response type mediated protection against SIV challenge. These findings implicate MHC-E-restricted CD8+ T cell responses as mediators of anti-SIV efficacy and indicate that translation of RhCMV/SIV vector efficacy to humans will likely require deletion of all the genes that inhibit these responses from the HCMV/HIV vector.One-sentence summaryEight genes in two spatially distinct RhCMV gene regions control induction of unconventionally restricted CD8+ T cell responses and the efficacy of RhCMV/SIV vaccine vectors against SIV challenge.

High salt diet (HS) decreases endothelial nitric oxide (NO) signaling as described in human and rodent studies. We have previously shown that inhibition of HDAC1 restores NO signaling in the afferent arteriole of HS fed rats. It remains unknown, however, how HS initiates endothelial dysfunction and activates HDAC1. We first investigated if HS induced endothelial dysfunction is acutely regulated by a circulating factor. To test this, male Sprague Dawley rats were fed normal salt diet (NS; 0.49% NaCl) or 2 weeks of HS (4.0% NaCl). Afferent arteriole vasoconstrictor responses to the NOS inhibitor, L-NAME, were monitored using the in vitro, blood perfused juxtamedullary nephron preparation. Perfusing arterioles from NS fed rats with HS blood blunted constriction to L-NAME (88 ± 1% of Con) compared to perfusing with NS blood (76 ± 2% of Con; P=0.0003), but not completely as seen in arterioles from HS fed rats perfused with HS blood (96 ± 2% of Con; P=0.0095). HS arterioles perfused with NS blood had similar L-NAME-induced constriction (75 ± 2% of Con) compared to arterioles from NS fed rats perfused with NS blood (P=0.9107). These results suggest HS induces endothelial function through an acute-acting circulating factor. This was not due to increased plasma arginase activity (NS: 7.96 ± 3.73 U/L vs HS: 5.42 ± 1.83 U/L P=0.5813) which competes with NO synthase 3 for arginine, nor was it due to decreased superoxide scavenging capacity of the plasma (NS: 7.96 ± 3.73 U/L vs HS: 5.42 ± 1.83 U/L P=0.5813) as measured by a cytochrome c reduction based assay. We then investigated if HS increased endothelial HDAC1 activity. Renal endothelial cells were isolated via magnetic activated cell sorting from NS and HS fed rats and incubated in plasma from the same rat. HDAC1 activity was monitored as the MS-275 (HDAC1 specific inhibitor) inhibitable portion of total HDAC activity. We found HS significantly increased renal endothelium HDAC1 activity (NS: 0.38 ± 0.03 pmol/min vs HS:0.94 ± 0.19 pmol/min, P=0.01). We conclude that HS disruption of renal microvascular NO signaling is initiated by a circulating factor(s) that is dependent upon increased endothelial HDAC1 activity.

Waterborne enteric viruses are an emerging cause of disease outbreaks and represent a major threat to global public health. Enteric viruses may originate from human wastewater and can undergo rapid transport through aquatic environments with minimal decay. Surveillance and source apportionment of enteric viruses in environmental waters is therefore essential for accurate risk management. However, individual monitoring of the >100 enteric viral strains that have been identified as aquatic contaminants is unfeasible. Instead, viral indicators are often used for quantitative assessments of wastewater contamination, viral decay and transport in water. An ideal indicator for tracking wastewater contamination should be (i) easy to detect and quantify, (ii) source-specific, (iii) resistant to wastewater treatment processes, and (iv) persistent in the aquatic environment, with similar behaviour to viral pathogens. Here, we conducted a comprehensive review of 127 peer-reviewed publications, to critically evaluate the effectiveness of several viral indicators of wastewater pollution, including common enteric viruses (mastadenoviruses, polyomaviruses, and Aichi viruses), the pepper mild mottle virus (PMMoV), and gut-associated bacteriophages (Type II/III FRNA phages and phages infecting human Bacteroides species, including crAssphage). Our analysis suggests that overall, human mastadenoviruses have the greatest potential to indicate contamination by domestic wastewater due to their easy detection, culturability, and high prevalence in wastewater and in the polluted environment. Aichi virus, crAssphage and PMMoV are also widely detected in wastewater and in the environment, and may be used as molecular markers for human-derived contamination. We conclude that viral indicators are suitable for the long-term monitoring of viral contamination in freshwater and marine environments and that these should be implemented within monitoring programmes to provide a holistic assessment of microbiological water quality and wastewater-based epidemiology, improve current risk management strategies and protect global human health., Graphical abstract Image 1, Highlights • Human mastadenoviruses are robust indicators for human-associated pollution in water. • Bacteroides-associated phages and crAssphage are promising indicators. • Multiple indicators should be used to assess wastewater treatment efficiency. • Survival and abundance of indicator viruses should be further assessed.

The effects of insulin on the vasculature remains a hotly debated and controversial topic. Its role on endothelial cells (ECs) is thought to be vasculoprotective, whereas its effects on vascular smooth muscle cells (SMCs) are mainly mitogenic according to in vitro data. We have previously demonstrated a suppressive effect of insulin treatment on neointimal growth in rodent models of restenosis under conditions of normal insulin sensitivity that was abolished in insulin resistant conditions. The objective of this study was to determine the endothelial-and SMC-specific effects of insulin treatment on neointimal growth in a murine model of restenosis. Mice were generated by crossing loxP-flanked insulin receptor (IR) mice with mice overexpressing tamoxifen-inducible Cre recombinase under the control of Tie2 (endothelial-specific receptor tyrosine kinase) or SMMHC (smooth muscle myosin heavy chain) promoters. Mice were fed with standard chow and implanted with insulin pellet (0.1 U/day) or sham (control) three days prior to undergoing femoral artery wire injury. Lastly, femoral arteries were collected for morphological analysis 28 days after wire injury. Tamoxifen-induced Tie2-Cre+ mice showed decreased IR expression in ECs compared with Tie2-Cre-, with no change in IR expression in SMCs. Neointimal area in Tie2-Cre- mice was reduced following insulin treatment, whereas it was unaffected in Tie2-Cre+ mice. Similarly, tamoxifen-induced SMMHC-Cre+ mice showed a reduction in IR expression in SMCs, but not in the ECs, compared to SMMHC-Cre- mice. In SMMHC-Cre- mice, neointimal area was decreased with insulin treatment; however, insulin surprisingly failed to have the same effects in SMMHC-Cre+ mice. These data demonstrate that insulin action in both ECs and SMCs is required for the antirestenotic effect of insulin in insulin sensitive conditions. Disclosure M. Gonzalez: None. Y. Mori: Research Support; Self; Taisho Pharmaceutical Co., Ltd. J. Guo: None. L.S. Dingwell: None. A. Giacca: None.

Viruses play a crucial and underexplored role in soil microbial ecosystems, but soil viral ecology has focused exclusively on DNA viruses. The role of RNA viruses in soil ecosystems has therefore been largely overlooked, despite their significant impact on public health and food security. Here, we report the first ever study to apply viromics to survey soil RNA viral communities from five sites along an altitudinal primary productivity gradient in the UK. We identified over 3,000 viral sequences, of which over half were unclassified, and newly identified viruses were placed in a global context by the phylogenetic comparison of their RNA-dependent RNA polymerase genes. Unlike DNA viral communities, the RNA viromes were heavily dominated by viruses of eukaryotes, including pathogens of plants, fungi, vertebrates and invertebrates. Sampling sites showed minimal similarity in viral community composition, suggesting that we have just scratched the surface of soil RNA viral diversity. Wider sequencing efforts and method development are required to further explore soil RNA viromes and understand their ecological function; however, this study represents an important step towards the characterisation of soil viral communities and interactions with their microbial hosts, which will provide a more holistic view of the biology of economically and ecologically important soils.

The development of maize (Zea mays L.) hybrids that are adapted to subtropical areas of North America should consider yield potential under heat and moisture stress, and reduced susceptibility to insect herbivory and disease. To aid in this process, maize hybrids (43 developmental and seven non-Bt commercial hybrids) were evaluated for severity of ear injury to Helicoverpa zea (Boddie) and Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), susceptibility to Aspergillus flavus (Link) (Deuteromycetes: Moniliales), and yield. In subtropical Corpus Christi and College Station, TX, field experiments conducted over three years revealed significant differences among maize hybrids with the rank of the selected measurements differing across the two locations. When the location by maize hybrid interaction was not significant, variation across the main factors of maize hybrid genetics (in all cases) and location (in some cases) was detected. In 2014, a significant location by maize hybrid interaction in yield but not aflatoxin and ear injury were likely associated with differences in weather between locations. In Corpus Christi in 2015, a location by maize hybrid interaction was detected for ear injury only. Overall, experimental maize hybrids, containing the inbred line Tx777, displayed partial resistance to insect derived ear injury in both locations, and some hybrid testcrosses exhibited low rates of aflatoxin accumulation while maintaining relatively high yields. Tx777 was selected from populations originating in Bolivia and adapted to subtropical climates. The most promising hybrid testcrosses had lower ear injury and aflatoxin accumulation, and good yield under varying heat and moisture stress at the two subtropical maize growing areas in this study.

Primary immunodeficiency (PID) is characterized by recurrent and often life-threatening infections, autoimmunity and cancer, and it poses major diagnostic and therapeutic challenges. Although the most severe forms of PID are identified in early childhood, most patients present in adulthood, typically with no apparent family history and a variable clinical phenotype of widespread immune dysregulation: about 25% of patients have autoimmune disease, allergy is prevalent and up to 10% develop lymphoid malignancies1–3. Consequently, in sporadic (or non-familial) PID genetic diagnosis is difficult and the role of genetics is not well defined. Here we address these challenges by performing whole-genome sequencing in a large PID cohort of 1,318 participants. An analysis of the coding regions of the genome in 886 index cases of PID found that disease-causing mutations in known genes that are implicated in monogenic PID occurred in 10.3% of these patients, and a Bayesian approach (BeviMed4) identified multiple new candidate PID-associated genes, including IVNS1ABP. We also examined the noncoding genome, and found deletions in regulatory regions that contribute to disease causation. In addition, we used a genome-wide association study to identify loci that are associated with PID, and found evidence for the colocalization of—and interplay between—novel high-penetrance monogenic variants and common variants (at the PTPN2 and SOCS1 loci). This begins to explain the contribution of common variants to the variable penetrance and phenotypic complexity that are observed in PID. Thus, using a cohort-based whole-genome-sequencing approach in the diagnosis of PID can increase diagnostic yield and further our understanding of the key pathways that influence immune responsiveness in humans. © 2020, The Author(s), under exclusive licence to Springer Nature Limited., Wellcome Trust, WT: 104807/Z/14/Z University College London, UCL: 100140/Z/12/Z, 107212/Z/15/Z, MR/L019027, 203141/Z/16/Z, 091157/Z/10/Z Bundesministerium für Bildung und Forschung, BMBF: 01ZX1306A University of Cambridge 201250/Z/16/Z 01ZX1709 Seventh Framework Programme, FP7 NIHR Bristol Biomedical Research Centre Deutsche Forschungsgemeinschaft, DFG Deutsche Forschungsgemeinschaft, DFG: EXC 2167-390884018 Juvenile Diabetes Research Foundation United Kingdom, JDRF: 9-2011-253, 5-SRA-2015-130-A-N National Institute for Health Research, NIHR: RG65966 Medical Research Council, MRC: RG95376, MR/L006197/1 Great Ormond Street Hospital for Children, GOSH, Acknowledgements The NBR-RD PID Consortium is part of the NIHR BioResource, for which funding was provided by the NIHR (NIHR, grant number RG65966). We acknowledge the participation of all NIHR BioResource volunteers, and thank the NIHR BioResource centre and staff for their contribution. J.E.D.T. is supported by the Medical Research Council (MRC) (RG95376 and MR/L006197/1); A.J.T. is supported by the Wellcome Trust (104807/Z/14/Z) and the NIHR Biomedical Research Centre at Great Ormond Street Hospital for Children NHS Foundation Trust and University College London; K.G.C.S. is supported by the MRC (programme grant MR/L019027) and is a Wellcome Investigator; A.J.C. was supported by the Wellcome Trust (091157/Z/10/Z, 107212/Z/15/Z, 100140/Z/12/Z, 203141/Z/16/Z), JDRF (9-2011-253, 5-SRA-2015-130-A-N), NIHR Oxford Biomedical Research Centre and NIHR Cambridge Biomedical Research Centre; E.E. has received funding from the European Union Seventh Framework Programme (FP7-PEOPLE-2013-COFUND) under grant agreement number 609020-Scientia Fellows; E.R. is supported by the Wellcome Trust (201250/Z/16/Z); D.E. is supported by the German Federal Ministry of Education and Research (BMBF) within the framework of the e:Med research and funding concept (SysInflame grant 01ZX1306A; GB-XMAP grant 01ZX1709) and funded by the Deutsche Forschungsgemeinschaft (DFG; German Research Foundation) under Germany’s Excellence Strategy (EXC 2167-390884018). The NIHR Cambridge Biomedical Research Centre (BRC) is a partnership between Cambridge University Hospitals NHS Foundation Trust and the University of Cambridge, funded by the NIHR. This research was co-funded by the support listed above and the NIHR Cambridge BRC.

Beta tropomyosin (Tpm2) is demonstrated for the first time at the protein level in a fish species, using a combination of electrophoresis, mass spectrometric peptide mapping and end-group analysis. Tpm2 accounts for 50% of the total tropomyosin in slow trunk muscle of the adult Atlantic salmon as determined by quantitative carboxypeptidase digestion and is also present in the head and pectoral fin. It is absent in the fast skeletal (lighter-toned) trunk muscle, the most abundant muscle, which is composed solely of an alpha-fast (Tpm1) isoform. In contrast to the mammalian homologues, salmon Tpm2 migrates faster than salmon Tpm1 in the presence of anionic detergent. Other distinguishing characteristics are a reduced content of cysteine (one per chain) and tyrosine (five per chain) and a unique carboxyl-terminal region (residues 276-284). Two isoforms (paralogs) of alpha-slow tropomyosin (Tpm3) having different contents of methionine and histidine exist in slow trunk muscle indicating duplication of the TPM3 gene. Minor skeletal muscles, surveyed for the first time, contain a mix of at least two tropomyosins - Tpm2 (~ 50% of total) in pectoral fin, jaw and tongue and another isoform, either Tpm1 (pectoral fin) or alpha-1-like Tpm (jaw and tongue). Cheek muscle contains Tpm1 and alpha 1-like Tpm in varying proportion depending upon the section (light or dark). Of the two tropomyosins in tongue, Tpm2 displays comparatively weaker affinity for troponin-Sepharose. A feature of the major sarcomeric tropomyosins in Atlantic salmon is a pair of neighbouring glycines situated between residues 20-90.

Pest and disease risk factors causing maize yield and kernel quality issues in subtropical areas of North America (between 35°N and 23.5°N latitude) are reviewed: preharvest Aspergillus flavus (Link) (Deuteromycetes: Moniliales) infection and propagation in maize ears and ear injury principally caused by Helicoverpa zea (Boddie) and Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae). Risk is affected by potentially interacting factors of maize genetics, ear feeding with insects, fungal inoculum and growth, and weather (rainfall, temperature, and humidity). This review gives special attention to integration of the most efficient pest and disease management strategy combinations and the potential interaction of insect ear feeding and A. flavus occurrence. Management strategies reviewed include maximizing partial genetic resistance to A. flavus and selection of appropriate hybrids for the area, biological control of A. flavus using non-aflatoxigenic strains, avoiding pests and diseases using cultural practices like early planting, and reducing H. zea and S. frugiperda ear injury using Bt transgenes. Understanding the combined influences and identifying combined management approaches may lead to reduced aflatoxin risk and maintaining yield. This review focuses on subtropical areas of North America because the amount of maize produced in warm environments within or similar to subtropical maize production areas is likely to increase due to projected increases in demand for maize and predicted increases in temperature may increase pest and disease risk.

BACKGROUND C-reactive protein (CRP) pharmacodynamic (PD) models have the potential to provide adjunctive methods for predicting the individual exposure response to antimicrobial therapy. We investigated CRP PD linked to a vancomycin pharmacokinetic (PK) model using routinely collected data from noncritical care adults in secondary care. METHODS Patients receiving intermittent intravenous vancomycin therapy in secondary care were identified. A 2-compartment vancomycin PK model was linked to a previously described PD model describing CRP response. PK and PD parameters were estimated using a Non-Parametric Adaptive Grid technique. Exposure-response relationships were explored with vancomycin area-under-the-concentration-time-curve (AUC) and EC50 (concentration of drug that causes a half maximal effect) using the index, AUC:EC50, fitted to CRP data using a sigmoidal Emax model. RESULTS Twenty-nine individuals were included. Median age was 62 (21-97) years. Fifteen (52%) patients were microbiology confirmed. PK and PD models were adequately fitted (r 0.83 and 0.82, respectively). There was a wide variation observed in individual Bayesian posterior EC50 estimates (6.95-48.55 mg/L), with mean (SD) AUC:EC50 of 31.46 (29.22). AUC:EC50 was fitted to terminal CRP with AUC:EC50 >19 associated with lower CRP value at 96-120 hours of therapy (100 mg/L versus 44 mg/L; P < 0.01). CONCLUSIONS The use of AUC:EC50 has the potential to provide in vivo organism and host response data as an adjunct for in vitro minimum inhibitory concentration data, which is currently used as the gold standard PD index for vancomycin therapy. This index can be estimated using routinely collected clinical data. Future work must investigate the role of AUC:EC50 in a prospective cohort and explore linkage with direct patient outcomes.

Global biodiversity loss is the cumulative result of local species declines. To combat biodiversity loss, detailed information on the temporal trends of at-risk species at local scales is needed. Here we report the results of a 13-year study of temporal change in bird occupancy in one of the most heavily modified biomes worldwide; the temperate woodlands of south-eastern Australia. We sought to determine if temporal changes in bird species were different between three broad native vegetation types (old-growth woodland, regrowth woodland and restoration plantings) and between species traits (body size, migratory status, rarity, woodland dependency, or diet). We found evidence of decline for over a quarter of all bird species for which we had sufficient data for detailed analysis (30 out of 108 species). In contrast, only 14 species increased significantly. Temporal change of birds was linked to life-history attributes, with patterns often being habitat-dependent. Nectarivores and large-bodied birds declined across all vegetation types, whereas small-bodied species increased, particularly in restoration plantings. Contrasting with patterns documented elsewhere, resident but not migratory species declined, with this trend strongest in restoration plantings. Finally, our analyses showed that, as a group, common birds tended to decline whereas rare birds tended to increase, with effects for both most pronounced in restoration plantings. Our results highlight the benefit of targeted restoration planting for some species, but also demonstrate that many common species that have long-persisted in human-dominated landscapes are experiencing severe declines.

The conventional approach to conserving threatened biota is to identify drivers of decline, instigate actions to mitigate threatening processes, and monitor interventions to test their effectiveness and ensure target species recover. In Australia, predation by introduced predators is a threatening process for many native mammals. Here we report the results of a 15 year monitoring study in an iconic Australian reserve, Booderee National Park, where exotic Red Fox (Vulpes vulpes) populations have been controlled through an intensive poison baiting program since 2003. Unexpectedly, we documented the collapse of native mammal fauna during this period, including fully arboreal species that should be largely unaffected by fox predation – such as the nationally Vulnerable Greater Glider (Petauroides volans) and Common Ringtail Possum (Pseudocheirus peregrinus). We used path analysis to explore potential causes of these unexpected declines. We found no compelling evidence to support hypotheses that competition with increasing native species, native predator release, or increases in native herbivores underpinned mammal declines. Beyond the path analysis, data from other studies completed both inside Booderee National Park and outside (where intensive fox baiting does not occur yet depleted fauna species remain), allowed us to rule out several drivers of change. The temporal declines we documented for arboreal marsupials were not anticipated nor explained by any clear mechanism. We propose the use of experimentally-guided reintroductions and translocations to: (1) restore empty niches such as the currently vacant apex mammal predator niche, (2) reconstruct the now depleted arboreal marsupial guild, and (3) further test key hypotheses associated with mammal decline. We also suggest that given the potential for perverse outcomes following large-scale management interventions (even those where there is high confidence of success), wildlife managers should consider maintaining reference areas (where there is no management intervention). Finally, as the declines we documented were unexpected and rapid, there is a clear need to develop more sensitive early warning signals to alert conservation managers to impending problems, allowing them to alter management regimes before major declines occur.

Increasing life expectancy is causing the prevalence of age-related diseases to rise, and there is an urgent need for new strategies to improve health at older ages. Reduced activity of insulin/insulin-like growth factor signaling (IIS) and mechanistic target of rapamycin (mTOR) nutrient-sensing signaling network can extend lifespan and improve health during aging in diverse organisms. However, the extensive feedback in this network and adverse side effects of inhibition imply that simultaneous targeting of specific effectors in the network may most effectively combat the effects of aging. We show that the mitogen-activated protein kinase kinase (MEK) inhibitor trametinib, the mTOR complex 1 (mTORC1) inhibitor rapamycin, and the glycogen synthase kinase-3 (GSK-3) inhibitor lithium act additively to increase longevity in Drosophila . Remarkably, the triple drug combination increased lifespan by 48%. Furthermore, the combination of lithium with rapamycin cancelled the latter’s effects on lipid metabolism. In conclusion, a polypharmacology approach of combining established, prolongevity drug inhibitors of specific nodes may be the most effective way to target the nutrient-sensing network to improve late-life health.

Background Weissella confusa, a Gram positive coccobacillus, is a rare cause of human disease. They are vancomycin resistant, bile aesculin positive, lactic acid fermenters previously classified as members of the Leuconostoc and Lactobacillus genera. Among the 19 recognised species of Weissella, W. confusa is most frequently associated with human infection, however as its name suggests establishing its pathogenicity and correctly identifying it using traditional phenotypic testing platforms can be challenging. Case report We present a case of W. confusa endocarditis in a male from Ghana where W. confusa is used in the fermenting process of commonly consumed food products. We discuss the microbiological diagnostic processes, including identification of this organism through use of matrix-assisted laser-desorption ionisation/time-of-flight (MALDI-ToF), and how the laboratory derived susceptibility tests were interpreted and applied to construct a successful therapeutic regime for infective endocarditis. Conclusions We review the literature around cases of invasive W. confusa disease, contextualising this organism in terms of relevance to patients who are immunocompromised or who have indwelling vascular access devices.

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection causes Coronavirus Disease 2019 (COVID-19), a pandemic that seriously threatens global health. SARS-CoV-2 propagates by packaging its RNA genome into membrane enclosures in host cells. The packaging of the viral genome into the nascent virion is mediated by the nucleocapsid (N) protein, but the underlying mechanism remains unclear. Here, we show that the N protein forms biomolecular condensates with viral genomic RNA both in vitro and in mammalian cells. While the N protein forms spherical assemblies with homopolymeric RNA substrates that do not form base pairing interactions, it forms asymmetric condensates with viral RNA strands. Cross-linking mass spectrometry (CLMS) identified a region that drives interactions between N proteins in condensates, and deletion of this region disrupts phase separation. We also identified small molecules that alter the size and shape of N protein condensates and inhibit the proliferation of SARS-CoV-2 in infected cells. These results suggest that the N protein may utilize biomolecular condensation to package the SARS-CoV-2 RNA genome into a viral particle., The packaging of the SARS-CoV-2 genome is mediated by the nucleocapsid (N) protein; this study shows that the N protein forms liquid condensates with viral genomic RNA and identifies small molecules that alter these condensates.

SARS-CoV-2 and the resulting COVID-19 pandemic represents one of the greatest recent threats to human health, wellbeing and economic growth. Wastewater-based epidemiology (WBE) of human viruses can be a useful tool for population-scale monitoring of SARS-CoV-2 prevalence and epidemiology to help prevent further spread of the disease, particularly within urban centres. Here we present a longitudinal analysis (March-July, 2020) of SARS-CoV-2 RNA prevalence in sewage across six major urban centres in the UK (total population equivalent 3 million) by q(RT-)PCR and viral genome sequencing. Our results demonstrate that levels of SARS-CoV-2 RNA generally correlated with the abundance of clinical cases recorded within the community in large urban centres, with a marked decline in SARS-CoV-2 RNA abundance following the implementation of lockdown measures. The strength of this association was weaker in areas with lower confirmed COVID-19 case numbers. Further sequencing analysis of SARS-CoV-2 from wastewater suggested that multiple genetically distinct clusters were co-circulating in the local populations covered by our sample sites, and that the genetic variants observed in wastewater reflected similar SNPs observed in contemporaneous samples from cases tested in clinical diagnostic laboratories. We demonstrate how WBE can be used for both community-level detection and tracking of SARS-CoV-2 and other virus’ prevalence, and can inform public health policy decisions. Although, greater understanding of the factors that affect SARS-CoV-2 RNA concentration in wastewater are needed for the full integration of WBE data into outbreak surveillance. In conclusion, our results lend support to the use of routine WBE for monitoring of SARS-CoV-2 and other human pathogenic viruses circulating in the population and assessment of the effectiveness of disease control measures., Graphical abstract Image, graphical abstract

The IFN gamma receptor 1 (IFNGR1) binds IFN-γ and activates gene transcription pathways crucial for controlling bacterial and viral infections. Although decreases in IFNGR1 surface levels have been demonstrated to inhibit IFN-γ signaling, little is known regarding the molecular mechanisms controlling receptor stability. Here, we show in epithelial and monocytic cell lines that IFNGR1 displays K48 polyubiquitination, is proteasomally degraded, and harbors three ubiquitin acceptor sites at K277, K279, and K285. Inhibition of glycogen synthase kinase 3 beta (GSK3β) destabilized IFNGR1 while overexpression of GSK3β increased receptor stability. We identified critical serine and threonine residues juxtaposed to ubiquitin acceptor sites that impacted IFNGR1 stability. In CRISPR–Cas9 IFNGR1 generated knockout cell lines, cellular expression of IFNGR1 plasmids encoding ubiquitin acceptor site mutations demonstrated significantly impaired STAT1 phosphorylation and decreased STAT1-dependent gene induction. Thus, IFNGR1 undergoes rapid site-specific polyubiquitination, a process modulated by GSK3β. Ubiquitination appears to be necessary for efficient IFNGR1-dependent gamma gene induction and represents a relatively uncharacterized regulatory mechanism for this receptor.

Biological control is known to be effective in reducing aflatoxin contamination of corn, and some transgenic corn hybrids incur reduced damage from corn earworm (Helicoverpa zea) than comparable non-Bt hybrids. We conducted seven field trials over two years to test the hypothesis that transgenic insect protection and biological control could be integrated to prevent mycotoxin contamination of corn. Corn hybrid N78N3111, expressing the highest degree of transgenic insect protection, was nearly 100 percent free from corn earworm damage and generally had less than half as much contamination from fumonisin compared to N78NGT, a near isogenic corn hybrid without insect protection. This insect protection, however, did not significantly prevent aflatoxin contamination. Soil application of non-aflatoxigenic biocontrol strains of Aspergillus flavus significantly reduced aflatoxin concentrations in corn. Biocontrol strain 21882 of A. flavus was especially effective and reduced aflatoxin contamination by about 90 percent over the seven field trials. There was no significant interaction between the insect protection and biocontrol treatments. Although no synergies were detected, the reduction of mycotoxins by both strategies supports application of both strategies in tandem. Economic factors external to the cost of the technologies will be a major determinant if the mycotoxin mitigation attained by use of these technologies will have a positive economic benefit.

In the absence of empirical evidence, invasive species are often assumed to have negative impacts because of their conspicuously high abundance. The giant African land snail Achatina (Lissachatina) fulica is one such invader where its impact in natural ecosystems remains completely untested. On Christmas Island (Indian Ocean), A. fulica has become established across large tracts of rainforest following the impacts of invasive yellow crazy ant (Anoplolepis gracilipes) in mutualism with non-native scale insects. Yellow crazy ants facilitate the secondary invasion of A. fulica by extirpating native red land crabs (Gecarcoidea natalis) that are normally effective predators of A. fulica. We used a multifaceted approach to investigate some potential impacts of abundant A. fulica in invaded rainforest. Over the course of a wet season, diel activity transects showed that A. fulica consumed detrital material almost exclusively. However, stable isotope analysis did not confidently identify A. fulica as a predominantly detritivorous species. We found no statistically significant treatment effects of A. fulica exclusion on standing leaf litter and seedling recruitment processes during a 6-month manipulative field study. However, litter cover and biomass did remain slightly higher where A. fulica were excluded, albeit with overlapping confidence intervals with control plots. Our study constitutes the first empirical test for impact of A. fulica in a natural ecosystem and suggests that for Christmas Island rainforest, this species is not a damaging invader. Other studies will need to assess the impacts of A. fulica in other natural areas before these findings could be considered broadly applicable.

This study was undertaken in part thanks to funding pro-vided by Biosis Research, Pty Ltd. Logistical suppor t wasprovided by staff at the Walpeup Research Station, ParksVictoria and the Ouyen Caravan Park.