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The Dendrobium hybrid cultivar ‘Emma White’ is an ornamental, successfully commercialised orchid. We used a gamma ray-induced early flowering mutant and the Illumina HiSeqX10 sequencing platform to generate the first draft de novo whole genome sequence and assembly. The draft sequence was 678,650,699 bp in length, comprising 447,500 contigs with an N50 of 1423 and 33.48% GC content. Comparing 95,529 predicted genes against the Uniprot database revealed 60,741 potential genes governing molecular functions, biological processes and cellular components. We identified 216,232 simple sequence repeats and 138,856 microsatellite markers. Chromosome-level genome assembly of Dendrobium huoshanense was used to RagTag-scaffold available contigs of the mutant, revealing a total length of 687,254,899 bp with an N50 of 2096. The longest final contiguous length was 18,000,059 bp from 30,571 bp. BUSCO genome completeness was 93.6%. This study is valuable for investigating the mechanisms of mutation, and developing Dendrobium hybrid cultivars using mutation breeding.

In vitro mutagenesis offers a feasible approach for developing new orchid cultivars through genetic manipulation. In the present study, protocorm-like bodies (PLBs) were exposed to gamma rays (10, 20, 40, 60, 80 Gy) to study in vitro growth responses and induction of mutants in Dendrobium ‘Emma White’. Both proliferation and regeneration of PLBs decreased progressively with increasing doses, except for a significantly enhanced growth response at 10 Gy. The optimal dose of gamma radiation for mutagenesis was found in the range 10 to 25 Gy based on the growth reduction curve. Analysis using a high-throughput cell analyzer revealed a significant reduction in nuclear DNA content at > 40 Gy doses. At 10 Gy treatment, the growth attributes, such as root length, plant height and leaf number, were significantly increased by 36%, 26% and 20%, respectively, compared to the control. This increase was significant over other tested doses as well. Testing of random amplified polymorphic DNA markers revealed the presence of detectable polymorphism among gamma mutant plantlets with a polymorphism information content value at 0.41. The gamma-ray-induced earliness in flower development was observed within 294 days post ex vitro growth of 10 Gy mutant compared to the control plants flowered after 959 days. Our results highlight the significance of gamma radiation in inducing enhanced growth, morphological variations and early floral initiation in Dendrobium, providing a basic framework for mutation breeding and improvement of orchids.

In vitro mutagenesis offers a feasible approach for developing new orchid cultivars through genetic manipulation. In the present study, protocorm-like bodies (PLBs) were exposed to gamma rays (10, 20, 40, 60, 80 Gy) to study in vitro growth responses and induction of mutants in Dendrobium ‘Emma White’. Both proliferation and regeneration of PLBs decreased progressively with increasing doses, except for a significantly enhanced growth response at 10 Gy. The optimal dose of gamma radiation for mutagenesis was found in the range 10 to 25 Gy based on the growth reduction curve. Analysis using a high-throughput cell analyzer revealed a significant reduction in nuclear DNA content at > 40 Gy doses. At 10 Gy treatment, the growth attributes, such as root length, plant height and leaf number, were significantly increased by 36%, 26% and 20%, respectively, compared to the control. This increase was significant over other tested doses as well. Testing of random amplified polymorphic DNA markers revealed the presence of detectable polymorphism among gamma mutant plantlets with a polymorphism information content value at 0.41. The gamma-ray-induced earliness in flower development was observed within 294 days post ex vitro growth of 10 Gy mutant compared to the control plants flowered after 959 days. Our results highlight the significance of gamma radiation in inducing enhanced growth, morphological variations and early floral initiation in Dendrobium, providing a basic framework for mutation breeding and improvement of orchids.

The Dendrobium hybrid cultivar 'Emma White' is an ornamental, successfully commercialised orchid. We used a gamma ray-induced early flowering mutant and the Illumina HiSeqX10 sequencing platform to generate the first draft de novo whole genome sequence and assembly. The draft sequence was 678,650,699 bp in length, comprising 447,500 contigs with an N50 of 1423 and 33.48% GC content. Comparing 95,529 predicted genes against the Uniprot database revealed 60,741 potential genes governing molecular functions, biological processes and cellular components. We identified 216,232 simple sequence repeats and 138,856 microsatellite markers. Chromosome-level genome assembly of Dendrobium huoshanense was used to RagTag-scaffold available contigs of the mutant, revealing a total length of 687,254,899 bp with an N50 of 2096. The longest final contiguous length was 18,000,059 bp from 30,571 bp. BUSCO genome completeness was 93.6%. This study is valuable for investigating the mechanisms of mutation, and developing Dendrobium hybrid cultivars using mutation breeding., Competing Interests: The authors declare that they have no competing interests., (© The Author(s) 2022.)

ABSTRACT: The objective of this study was to evaluate the effect of a novel consensus bacterial 6-phytase variant (PhyG) on egg productivity, eggshell quality, and body composition of laying hens fed inorganic phosphate-free diets with reduced energy and nutrients from 23 to 72 wk of age. Five treatments were randomly assigned, performing 28 replicates per treatment with 4 hens each, totaling 560 Hy-Line W80 birds. A positive control (PC) feed was formulated to contain adequate levels of energy and nutrients. A negative control (NC) feed was formulated without added inorganic phosphate (0.12% nonphytic phosphorus [nPP]) and reduced in Ca, Na, dig AA, and metabolizable energy in comparison with PC feed. Phytase was supplemented in the NC feed at 0, 300, 600, and 900 FTU/kg of feed. The responses evaluated were performance, egg quality, economic analysis, body composition, and tibia composition. Data were analyzed by a 2-factor (diet and age) repeated measure analysis. Overall, the feed intake, hen-day egg production, egg mass, and egg revenue were reduced by the complete removal of dicalcium phosphate (DCP) (P < 0.05). Supplement phytase in the NC diet elicits a positive response on each one of those variables. Laying hens consuming the NC feed with 900 FTU/kg of phytase produced more eggs per hen-housed compared with the phytase dosages of 300 and 600 FTU/kg. Body composition was not affected by dietary nPP, Ca, Na, dig AA, and energy reductions (P > 0.05). At 72-wk-old, tibia ash was reduced in hens consuming the NC diet vs. PC (P < 0.05) and no difference was observed between hens supplemented with phytase and the PC feed. Margin over feeding cost increased in a dose-dependent manner with phytase supplementation. Supplementation with 900 FTU/kg of phytase is recommended to improve the number of eggs produced per hen-housed and the number of marketable eggs produced through 23 to 72 wk of age, under this dietary setting.

'CLIMATE CHANGE is the most severe problem we are facing today--more serious even than the threat of terrorism,' according to the government's chief scientist, David King. This might sound alarmist, [...]

BackgroundClinical utility of routinely measured serial biomarkers in predicting escalation of inpatient care intensity and mortality among hospitalized patients with COVID-19 remains unknown.MethodsThis retrospective cohort study included patients with COVID-19 who admitted to the Massachusetts General Hospital between March and June 2020 and January to March 2021. White blood cell (WBC) count, platelet count, C-reactive protein (CRP), and D-dimer values were measured on days 1, 3, and 7 of admission. Clinical outcomes include 30- and 60-day morality, ICU transfer, and overall survival (OS) over a follow-up period of 90 days. The association between serial biomarkers and outcomes were assessed using multivariable logistic regression and Cox proportional hazards models.Measurements and main resultsOf the 456 patients hospitalized with COVID-19, 199 (43.6%) were ICU, 179 (39.3%) were medical floor, and 78 (17.1%) were initially admitted to the medical floor and then transferred to the ICU. In adjusted analyses, each unit increase in the slope of CRP was associated with a 42% higher odds of ICU transfer after controlling for the initial admission level (OR = 1.42, 95% CI: 1.25-1.65, P < 0.001). Including serial change in CRP levels from initial level on admission achieved the greatest predictive accuracy for ICU transfer (AUC = 0.72, 95% CI: 0.64-0.79).ConclusionsSerial change in CRP levels from admission is associated with escalations of inpatient care intensity and mortality among hospitalized patients with COVID-19.

Integrated approaches to testing and assessments (IATAs) have been proposed as a method to organise new approach methodologies in order to replace traditional animal testing for chemical safety assessments. To capture the mechanistic aspects of toxicity assessments, IATAs can be framed around the adverse outcome pathway (AOP) concept. To utilise AOPs fully in this context, a sufficient number of pathways need to be present to develop fit for purpose IATAs. In silico approaches can support IATA through the provision of predictive models and also through data integration to derive conclusions using a weight-of-evidence approach. To examine the maturity of a developmental and reproductive toxicity (DART) AOP network derived from the literature, an assessment of its coverage was performed against a novel toxicity dataset. A dataset of diverse compounds, with data from studies performed according to OECD test guidelines TG-421 and TG-422, was curated to test the performance of an in silico model based on the AOP network – allowing for the identification of knowledge gaps within the network. One such gap in the knowledge was filled through the development of an AOP stemming from the molecular initiating event ‘glutathione reaction with an electrophile’ leading to male fertility toxicity. The creation of the AOP provided the mechanistic rationale for the curation of pre-existing structural alerts to relevant key events. Integrating this new knowledge and associated alerts into the DART AOP network will improve its coverage of DART-relevant chemical space. In addition, broadening the coverage of AOPs for a particular regulatory endpoint may facilitate the development of, and confidence in, robust IATAs.

Atlantic herring in International Council for Exploration of the Sea (ICES) Divisions 6.a, 7.b–c comprises at least three populations, distinguished by temporal and spatial differences in spawning, which have until recently been managed as two stocks defined by geographical delineators. Outside of spawning the populations form mixed aggregations, which are the subject of acoustic surveys. The inability to distinguish the populations has prevented the development of separate survey indices and separate stock assessments. A panel of 45 single-nucleotide polymorphisms, derived from whole-genome sequencing, were used to genotype 3480 baseline spawning samples (2014–2021). A temporally stable baseline comprising 2316 herring from populations known to inhabit Division 6.a was used to develop a genetic assignment method, with a self-assignment accuracy greater than 90%. The long-term temporal stability of the assignment model was validated by assigning archive (2003–2004) baseline samples (270 individuals) with a high level of accuracy. Assignment of non-baseline samples (1514 individuals) from Divisions 6.a, 7.b–c indicated previously unrecognized levels of mixing of populations outside of the spawning season. The genetic markers and assignment models presented constitute a ‘toolbox’ that can be used for the assignment of herring caught in mixed survey and commercial catches in Division 6.a into their population of origin with a high level of accuracy.

Mechanically alloyed Fe-based alloys with oxide dispersion strengthening have largely dropped out of the marketplace due to high cost related to problems with complex and unreliable processing. Nevertheless, the desirable properties of oxide dispersion strengthened (ODS) steels have motivated research on alternate processing routes aimed at improving processing simplicity and reliability. Powders produced by gas atomization reaction synthesis (GARS) consist of stable Fe-Y intermetallic phases and a Cr surface oxide layer that acts as a chemical reservoir during solid-state processing and heat treatment to form a high density of nano-scale oxides. This research explores the use of Fe GARS powders, with 15 wt% Cr with micro-alloyed additions of 0.15 wt% Y and 0.10% Ti, in laser powder bed fusion (LPBF) additive manufacturing (AM), and evaluates the effectiveness of oxide dispersoid formation in the liquid melt pool. Additional oxygen was introduced by varying the LPBF chamber atmospheres using Ar, Ar + 1 wt% O, Ar + 5 wt% O, and air. Characterization of LPBF consolidated solids demonstrated the formation of a high density of nano-scale Y-Ti oxides in the build microstructures from the GARS precursor powders.

Among other tests, Barts Health NHS Trust clinical transplantation laboratory conducts two important gene-detection tests: human leucocyte antigen (HLA)-B*27 (‘B27’, associated with the diagnosis of ankylosing spondylitis) and HLA-B*57:01 (‘B57’, associated with prediction of abacavir hypersensitivity disorder). The turnaround time (TaT) from sample receipt to return of results is important to clinicians and their patients but was not monitored. Furthermore, we anticipated an imminent increase in demand from a forthcoming pathology service merger, together with long-term increases with the rise of personalised genetic medicine.In this quality improvement project, we identified current TaT performance and sources of delay. Over three plan-do-study-act (PDSA) cycles, we tested three change ideas, two involving using IT to remove manual administrative steps and alert us to samples needing progressing; both were retained. The other change involved separating out the targeted tests; we judged this not worthwhile with current demand levels, although something to be re-examined when volumes increase. During the project, we reduced mean TaT from 3.8 to 3.3 days and increased the proportion within our 5-day target from 78% to 100%. These have been sustained (at 3.4 days and 97%) for the 3 months following our PDSA cycles and illustrate that reducing variation can be as impactful as reducing the mean.We conducted this project during the COVID-19 disruption, which reduced demand substantially. We took advantage of this to allow staff to spend time on these improvement activities. Another interesting feature of the work is that during the project, we compared changes in performance on our targeted B27/B57 tests with that on another comparable test as a control, to consider the impact of the general increased attention (the Hawthorne effect). We found that performance on this control also increased comparably, but then fell away after our project finished, while it did not for B27/B57.

IT MAY have taken two and a half years to get approval, but the Winfield clinic, Queen Elizabeth University Hospital, is now a success story of which Hilary Jefferies can [...]

Abstract Objective Obesity is associated with a high incidence of obstructive sleep apnoea (OSA). Bariatric surgery is postulated to lead to OSA resolution, but there is inconclusive evidence on its efficacy. We used objective measurements to determine the rate of resolution or improvement of OSA in patients who had bariatric procedures in our unit. Results Data was analysed on all patients with OSA who underwent bariatric procedures [laparoscopic Roux-en-Y gastric bypass (LRYGB) and sleeve gastrectomy (LSG)] between June 2012 and September 2016 in our unit. 47 patients (26.7%) were diagnosed with OSA. Mean age was 48.5 years. 63.8% were female. 43 required nocturnal continuous positive airway pressure (CPAP) support. Procedures were LRYGB (n = 26) and LSG (n = 21). Mean excess weight loss was 56.1%. Mean start apnoea-hypopnoea index (AHI) on CPAP was 6.4 events/hr and end AHI was 1.4 events/h. 14 patients (32.6%) had complete OSA resolution and 12 (27.9%) showed improvement in pressure support requirements. We demonstrated that 55.3% of patients had resolution or improvement in OSA following bariatric surgery. However, there was a high rate of non-attendance of follow-up appointments. Future efforts will involve analysis of the reasons for this to ensure more robust monitoring.

Purpose: With the increasing burden of chronic pain and opioid use, provider shortages in Eastern Kentucky and West Virginia have experienced many challenges related to chronic pain management. This study tested a practice facilitator model in both academic and community clinics that selected and implemented best practice processes to better assist patients with chronic pain and increase the use of interdisciplinary health care services. Methods: Using a quasi-experimental design, a practice facilitator was assigned to each state’s clinics and trained clinic teams in quality improvement methods to implement chronic pain tool(s) and workflow processes. Charts for 695 patients with chronic pain using opioids, from 8 randomly selected clinics in eastern Appalachia, were reviewed to assess for changes in clinic processes. Results: Statistically significant improvements were found in 10 out of 16 chronic pain best practice process measures. These included improved workflow implementation (P < 0.001), increased urine drug screen test orders (P = 0.001) and increased utilization of controlled medication agreements (P = 0.004). In total, 7 of 8 clinics significantly improved in at least one, if not all, selected and implemented process measures. Conclusions: Our findings indicate that practice facilitation, standardization of workflows and formation of structured clinical teams can improve processes of care in chronic pain management and facilitate the use of interdisciplinary services. Future studies are needed to assess long-term patient-centered outcomes that may result from improved processes of chronic pain care.