Increased expression of a microRNA correlates with anthelmintic resistance in parasitic nematodes

Resistance to anthelmintic drugs is a major problem in the global fight against parasitic nematodes infecting humans and animals. While previous studies have identified mutations in drug target genes in resistant parasites, changes in the expression levels of both targets and transporters have also been reported. The mechanisms underlying these changes in gene expression are unresolved. Here, we take a novel approach to this problem by investigating the role of small regulatory RNAs in drug resistant strains of the important parasite Haemonchus contortus. microRNAs (miRNAs) are small (22 nt) non-coding RNAs that regulate gene expression by binding predominantly to the 3′ UTR of mRNAs. Changes in miRNA expression have been implicated in drug resistance in a variety of tumor cells. In this study, we focused on two geographically distinct ivermectin resistant strains of H. contortus and two lines generated by multiple rounds of backcrossing between susceptible and resistant parents, with ivermectin selection. All four resistant strains showed significantly increased expression of a single miRNA, hco-miR-9551, compared to the susceptible strain. This same miRNA is also upregulated in a multi-drug-resistant strain of the related nematode Teladorsagia circumcincta. hco-miR-9551 is enriched in female worms, is likely to be located on the X chromosome and is restricted to clade V parasitic nematodes. Genes containing predicted binding sites for hco-miR-9551 were identified computationally and refined based on differential expression in a transcriptomic dataset prepared from the same drug resistant and susceptible strains. This analysis identified three putative target mRNAs, one of which, a CHAC domain containing protein, is located in a region of the H. contortus genome introgressed from the resistant parent. hco-miR-9551 was shown to interact with the 3′ UTR of this gene by dual luciferase assay. This study is the first to suggest a role for miRNAs and the genes they regulate in drug resistant parasitic nematodes. miR-9551 also has potential as a biomarker of resistance in different nematode species

Efficacy of pre- and post-emergence herbicide combinations on weed control in no-till mechanically transplanted rice

No-till mechanized-transplanted rice was evaluated for different combinations of pre- and post-emergence herbicides to determine feasible, economically viable weed management options to control complex weed flora in rice fields. All pre-emergence herbicides significantly reduced the population of grassy weeds; of these, pendimethalin resulted in the greatest reductions (83%) at 15 days after transplanting (DAT). Among five post-emergence herbicide treatments, the combination of bispyribac-sodium (10%SP) + pyrazosulfuron (10%WP) was found to be the most effective in controlling all weed flora at both 35 and 55 DAT. The sequential application of pendimethalin (pre-emergence) followed bispyribac-sodium + pyrazosulfuron (post-emergence) resulted in significantly higher rice grain yield (4.4 t-ha−1) and relative gross-margin (417 USD-ha−1) than all other treatments. A strong negative correlation was observed between rice grain yield and weed biomass, and a strong positive correlation between rice grain yield and weed control efficiency. Our findings demonstrate the potential to combine pre- and post-emergence herbicides in no-till mechanized-transplanted rice; these findings have applications globally in regions where rice is established by no-till or mechanized transplanting

Minerals and chelated-based manganese fertilization influences the productivity, uptake, and mobilization of manganese in wheat (Triticum aestivum L.) in sandy loam soils

Manganese (Mn) is an essential micronutrient in plants, and it is necessary for hydrolysis in photosystem II, chlorophyll biosynthesis, and also chloroplast breakdown. Limited Mn availability in light soil resulted in interveinal chlorosis, poor root development, and the development of fewer tillers, particularly staple cereals including wheat, while foliar Mn fertilizers were found efficient in improving crop yield as well as Mn use efficiency. In the above context, a study was conducted in consecutive two wheat growing seasons for screening of the most effective and economical Mn treatment for improving the yield and Mn uptake in wheat and to compare the relative effectiveness of MnCO3 against the recommended dose of MnSO4 for wheat. To fulfill the aims of the study, three manganese products, namely, 1) manganese carbonate MnCO3 (26% Mn w/w and 3.3% N w/w), 2) 0.5% MnSO4·H2O (30.5% Mn), and 3) Mn-EDTA solution (12% Mn), were used as experimental treatments. Treatments and their combinations were as follows: two levels of MnCO3 (26% Mn) @ 750 and 1,250 ml ha−1 were applied at the two stages (i.e., 25–30 and 35–40 days after sowing) of wheat, and three sprays each of 0.5% MnSO4 (30.5% Mn) and Mn-EDTA (12% Mn) solution were applied in other plots. The 2-year study showed that Mn application significantly increased the plant height, productive tillers plant−1, and 1,000 grain weight irrespective of fertilizer source. The results of MnSO4 for grain yield wheat as well as uptake of Mn were statistically at par with both levels (750 and 1,250 ml ha−1) of MnCO3 with two sprays at two stages of wheat. However, the application of Mn in the form of 0.5% MnSO4·H2O (30.5% Mn) was found more economical than MnCO3, while the mobilization efficiency index (1.56) was found maximum when Mn was applied in MnCO3 with two sprays (750 and 1,250 ml ha−1) in the two stages of wheat. Thus, the present study revealed that MnCO3 can be used as an alternative to MnSO4 to enhance the yield and Mn uptake of wheat

Growth, nodulation, yield, nitrogen uptake, and economics of lentil as influenced by sowing time, tillage, and management practices

Crop management practices and variety are two very important parameters that decides the crop performance. A field experiment was carried out during the two consecutive rabi seasons of 2018–19 and 2019–20 to determine the impact of sowing timing, tillage operation, and variety on the growth, development, yield characteristics, and nitrogen uptake in lentil crops. The experiment was conducted in a split-split plot design with 3 replications comprising two different sowing conditions (S1: early sowing after harvesting of short duration kharif rice, S2: delayed sowing after harvesting of long duration kharif rice) in main plots, three different tillage operations (T1: Relay cropping, T2: Zero tillage, T3: Conventional tillage) in subplots and two different varieties (V1: short duration: L4717, V2: long period: Moitri) in subplots. The findings demonstrated a substantial interaction between sowing time, tillage, and variety on various growth and yield parameters of lentil crops. The early sowing of lentil crops (early November) yielded 4.8% more (1,105 kg ha−1) than late November sowing and adapting to the short-duration variety L4717 over the long-duration cultivar Moitri resulted in a yield increase of 5.9% (1,086 kg ha−1). Apart from providing a higher yield, it also provided an opportunity to take another crop like leafy vegetables. Among the three tillage practices adopted, conventional tillage produced the lowest yield (1,017 kg ha−1) in both experimental years. In contrast, a yield increase of 6.9% and 26.9% in relay cropping and zero tillage systems was observed, respectively. Early-sown lentils with no-tillage and a short-duration variety reached a certain phenophase faster than other combinations (life cycle: 96.2 and 98.7 days for lentils in both years). For both the sowing times, the growth parameters and the number of nodules plant−1 were highly correlated with nitrogen uptake at different stages of the life cycle. High net returns (Rs. 51,220 and 59,257) leading to higher benefit-cost ratios were observed under the treatment combination of early sowing + zero tillage + short duration variety. Therefore, the study found that short-duration lentil cultivars in combination with early sowing in the zero-tillage system are the best agronomic approach for the sustainability of lentil production after the monsoon rice harvest

PreImplantation Trial of Histopathology In renal Allografts (PITHIA): a stepped-wedge cluster randomised controlled trial protocol.

INTRODUCTION: Most potential kidney transplant donors in the UK are aged over 60 years, yet increasing donor age is associated with poorer graft survival and function. Urgent preimplantation kidney biopsy can identify chronic injury, and may aid selection of better 'quality' kidneys from this group. However, the impact of biopsy on transplant numbers remains unproven. The PreImplantation Trial of Histopathology In renal Allografts (PITHIA) study will assess whether the introduction of a national, 24 hours, digital histopathology service increases the number, and improves outcomes, of kidneys transplanted in the UK from older deceased donors. METHODS AND ANALYSIS: PITHIA is an open, multicentre, stepped-wedge cluster randomised study, involving all UK adult kidney transplant centres. At 4-monthly intervals, a group of 4-5 randomly selected clusters (transplant centres) will be given access to remote, urgent, digital histopathology (total intervention period, 24 months). The trial has two primary end points: it is powered for an 11% increase in the proportion of primary kidney offers from deceased donors aged over 60 years that are transplanted, and a 6 mL/min increase in the estimated glomerular filtration rate of recipients at 12 months post-transplant. This would equate to an additional 120 kidney transplants performed in the UK annually. Trial outcome data will be collected centrally via the UK Transplant Registry held by NHS Blood and Transplant (NHSBT) and will be analysed using mixed effects models allowing for clustering within centres and adjusting for secular trends. An accompanying economic evaluation will estimate the cost-effectiveness of the service to the National Health Service. ETHICS AND DISSEMINATION: The study has been given favourable ethical opinion by the Cambridge South Research Ethics Committee and is approved by the Health Research Authority. We will present our findings at key transplant meetings, publish results within 4 years of the trial commencing and support volunteers at renal patient groups to disseminate the trial outcome. TRIAL REGISTRATION NUMBER: ISRCTN11708741; Pre-results.nih

A new approach to treatment of resistant gram-positive infections: potential impact of targeted IV to oral switch on length of stay

BACKGROUND: Patients prescribed intravenous (IV) glycopeptides usually remain in hospital until completion of this treatment. Some of these patients could be discharged earlier if a switch to an oral antibiotic was made. This study was designed to identify the percentage of inpatients currently prescribed IV glycopeptides who could be discharged earlier if a switch to an oral agent was used, and to estimate the number of bed days that could be saved. We also aimed to identify the patient group(s) most likely to benefit, and to estimate the number of days of IV therapy that could be prevented in patients who remained in hospital. METHODS: Patients were included if they were prescribed an IV glycopeptide for 5 days or more. Predetermined IV to oral antibiotic switch criteria and discharge criteria were applied. A multiple logistic regression model was used to identify the characteristics of the patients most likely to be suitable for earlier discharge. RESULTS: Of 211 patients, 62 (29%) could have had a reduced length of stay if they were treated with a suitable oral antibiotic. This would have saved a total of 649 inpatient days (median 5 per patient; range 1–54). A further 31 patients (15%) could have switched to oral therapy as an inpatient thus avoiding IV line use. The patients most likely to be suitable for early discharge were those with skin and soft tissue infection, under the cardiology, cardiothoracic surgery, orthopaedics, general medical, plastic surgery and vascular specialities, with no high risk comorbidity and less than five other regularly prescribed drugs. CONCLUSION: The need for glycopeptide therapy has a significant impact on length of stay. Effective targeting of oral antimicrobials could reduce the need for IV access, allow outpatient treatment and thus reduce the length of stay in patients with infections caused by antibiotic resistant gram-positive bacteria

Gemini GMOS and WHT SAURON integral-field spectrograph observations of the AGN driven outflow in NGC 1266

We use the SAURON and GMOS integral field spectrographs to observe the active galactic nucleus (AGN) powered outflow in NGC 1266. This unusual galaxy is relatively nearby (D=30 Mpc), allowing us to investigate the process of AGN feedback in action. We present maps of the kinematics and line strengths of the ionised gas emission lines Halpha, Hbeta, [OIII], [OI], [NII] and [SII], and report on the detection of Sodium D absorption. We use these tracers to explore the structure of the source, derive the ionised and atomic gas kinematics and investigate the gas excitation and physical conditions. NGC 1266 contains two ionised gas components along most lines of sight, tracing the ongoing outflow and a component closer to the galaxy systemic, the origin of which is unclear. This gas appears to be disturbed by a nascent AGN jet. We confirm that the outflow in NGC 1266 is truly multiphase, containing radio plasma, atomic, molecular and ionised gas and X-ray emitting plasma. The outflow has velocities up to \pm900 km/s away from the systemic velocity, and is very likely to be removing significant amounts of cold gas from the galaxy. The LINER-like line-emission in NGC 1266 is extended, and likely arises from fast shocks caused by the interaction of the radio jet with the ISM. These shocks have velocities of up to 800 km/s, which match well with the observed velocity of the outflow. Sodium D equivalent width profiles are used to set constraints on the size and orientation of the outflow. The ionised gas morphology correlates with the nascent radio jets observed in 1.4 GHz and 5 GHz continuum emission, supporting the suggestion that an AGN jet is providing the energy required to drive the outflow.Comment: Contains 18 figures. Accepted to MNRA

Introgression of Ivermectin Resistance Genes into a Susceptible Haemonchus contortus Strain by Multiple Backcrossing

Anthelmintic drug resistance in livestock parasites is already widespread and in recent years there has been an increasing level of anthelmintic drug selection pressure applied to parasitic nematode populations in humans leading to concerns regarding the emergence of resistance. However, most parasitic nematodes, particularly those of humans, are difficult experimental subjects making mechanistic studies of drug resistance extremely difficult. The small ruminant parasitic nematode Haemonchus contortus is a more amenable model system to study many aspects of parasite biology and investigate the basic mechanisms and genetics of anthelmintic drug resistance. Here we report the successful introgression of ivermectin resistance genes from two independent ivermectin resistant strains, MHco4(WRS) and MHco10(CAVR), into the susceptible genome reference strain MHco3(ISE) using a backcrossing approach. A panel of microsatellite markers were used to monitor the procedure. We demonstrated that after four rounds of backcrossing, worms that were phenotypically resistant to ivermectin had a similar genetic background to the susceptible reference strain based on the bulk genotyping with 18 microsatellite loci and individual genotyping with a sub-panel of 9 microsatellite loci. In addition, a single marker, Hcms8a20, showed evidence of genetic linkage to an ivermectin resistance-conferring locus providing a starting point for more detailed studies of this genomic region to identify the causal mutation(s). This work presents a novel genetic approach to study anthelmintic resistance and provides a “proof-of-concept” of the use of forward genetics in an important model strongylid parasite of relevance to human hookworms. The resulting strains provide valuable resources for candidate gene studies, whole genome approaches and for further genetic analysis to identify ivermectin resistance loci

Addition of docetaxel, zoledronic acid, or both to first-line long-term hormone therapy in prostate cancer (STAMPEDE): Survival results from an adaptive, multiarm, multistage, platform randomised controlled trial

BACKGROUND Long-term hormone therapy has been the standard of care for advanced prostate cancer since the 1940s. STAMPEDE is a randomised controlled trial using a multiarm, multistage platform design. It recruits men with high-risk, locally advanced, metastatic or recurrent prostate cancer who are starting first-line long-term hormone therapy. We report primary survival results for three research comparisons testing the addition of zoledronic acid, docetaxel, or their combination to standard of care versus standard of care alone. METHODS Standard of care was hormone therapy for at least 2 years; radiotherapy was encouraged for men with N0M0 disease to November, 2011, then mandated; radiotherapy was optional for men with node-positive non-metastatic (N+M0) disease. Stratified randomisation (via minimisation) allocated men 2:1:1:1 to standard of care only (SOC-only; control), standard of care plus zoledronic acid (SOC + ZA), standard of care plus docetaxel (SOC + Doc), or standard of care with both zoledronic acid and docetaxel (SOC + ZA + Doc). Zoledronic acid (4 mg) was given for six 3-weekly cycles, then 4-weekly until 2 years, and docetaxel (75 mg/m(2)) for six 3-weekly cycles with prednisolone 10 mg daily. There was no blinding to treatment allocation. The primary outcome measure was overall survival. Pairwise comparisons of research versus control had 90% power at 2·5% one-sided α for hazard ratio (HR) 0·75, requiring roughly 400 control arm deaths. Statistical analyses were undertaken with standard log-rank-type methods for time-to-event data, with hazard ratios (HRs) and 95% CIs derived from adjusted Cox models. This trial is registered at ClinicalTrials.gov (NCT00268476) and ControlledTrials.com (ISRCTN78818544). FINDINGS 2962 men were randomly assigned to four groups between Oct 5, 2005, and March 31, 2013. Median age was 65 years (IQR 60-71). 1817 (61%) men had M+ disease, 448 (15%) had N+/X M0, and 697 (24%) had N0M0. 165 (6%) men were previously treated with local therapy, and median prostate-specific antigen was 65 ng/mL (IQR 23-184). Median follow-up was 43 months (IQR 30-60). There were 415 deaths in the control group (347 [84%] prostate cancer). Median overall survival was 71 months (IQR 32 to not reached) for SOC-only, not reached (32 to not reached) for SOC + ZA (HR 0·94, 95% CI 0·79-1·11; p=0·450), 81 months (41 to not reached) for SOC + Doc (0·78, 0·66-0·93; p=0·006), and 76 months (39 to not reached) for SOC + ZA + Doc (0·82, 0·69-0·97; p=0·022). There was no evidence of heterogeneity in treatment effect (for any of the treatments) across prespecified subsets. Grade 3-5 adverse events were reported for 399 (32%) patients receiving SOC, 197 (32%) receiving SOC + ZA, 288 (52%) receiving SOC + Doc, and 269 (52%) receiving SOC + ZA + Doc. INTERPRETATION Zoledronic acid showed no evidence of survival improvement and should not be part of standard of care for this population. Docetaxel chemotherapy, given at the time of long-term hormone therapy initiation, showed evidence of improved survival accompanied by an increase in adverse events. Docetaxel treatment should become part of standard of care for adequately fit men commencing long-term hormone therapy. FUNDING Cancer Research UK, Medical Research Council, Novartis, Sanofi-Aventis, Pfizer, Janssen, Astellas, NIHR Clinical Research Network, Swiss Group for Clinical Cancer Research

SARS-CoV-2 lineage B.1.1.7 is associated with greater disease severity among hospitalised women but not men: multicentre cohort study.

BACKGROUND: SARS-CoV-2 lineage B.1.1.7 has been associated with an increased rate of transmission and disease severity among subjects testing positive in the community. Its impact on hospitalised patients is less well documented. METHODS: We collected viral sequences and clinical data of patients admitted with SARS-CoV-2 and hospital-onset COVID-19 infections (HOCIs), sampled 16 November 2020 to 10 January 2021, from eight hospitals participating in the COG-UK-HOCI study. Associations between the variant and the outcomes of all-cause mortality and intensive therapy unit (ITU) admission were evaluated using mixed effects Cox models adjusted by age, sex, comorbidities, care home residence, pregnancy and ethnicity. FINDINGS: Sequences were obtained from 2341 inpatients (HOCI cases=786) and analysis of clinical outcomes was carried out in 2147 inpatients with all data available. The HR for mortality of B.1.1.7 compared with other lineages was 1.01 (95% CI 0.79 to 1.28, p=0.94) and for ITU admission was 1.01 (95% CI 0.75 to 1.37, p=0.96). Analysis of sex-specific effects of B.1.1.7 identified increased risk of mortality (HR 1.30, 95% CI 0.95 to 1.78, p=0.096) and ITU admission (HR 1.82, 95% CI 1.15 to 2.90, p=0.011) in females infected with the variant but not males (mortality HR 0.82, 95% CI 0.61 to 1.10, p=0.177; ITU HR 0.74, 95% CI 0.52 to 1.04, p=0.086). INTERPRETATION: In common with smaller studies of patients hospitalised with SARS-CoV-2, we did not find an overall increase in mortality or ITU admission associated with B.1.1.7 compared with other lineages. However, women with B.1.1.7 may be at an increased risk of admission to intensive care and at modestly increased risk of mortality.This report was produced by members of the COG-UK-HOCI Variant substudy consortium. COG-UK-HOCI is part of COG-UK. COG-UK is supported by funding from the Medical Research Council (MRC) part of UK Research & Innovation (UKRI), the National Institute of Health Research (NIHR) and Genome Research Limited, operating as the Wellcome Sanger Institute