An In Vitro Pipeline for Screening and Selection of Citrus-Associated Microbiota with Potential Anti-"Candidatus Liberibacter asiaticus" Properties.

Huanglongbing (HLB) is a destructive citrus disease that is lethal to all commercial citrus plants, making it the most serious citrus disease and one of the most serious plant diseases. Because of the severity of HLB and the paucity of effective control measures, we structured this study to encompass the entirety of the citrus microbiome and the chemistries associated with that microbial community. We describe the spatial niche diversity of bacteria and fungi associated with citrus roots, stems, and leaves using traditional microbial culturing integrated with culture-independent methods. Using the culturable sector of the citrus microbiome, we created a microbial repository using a high-throughput bulk culturing and microbial identification pipeline. We integrated an in vitro agar diffusion inhibition bioassay into our culturing pipeline that queried the repository for antimicrobial activity against Liberibacter crescens, a culturable surrogate for the nonculturable "Candidatus Liberibacter asiaticus" bacterium associated with HLB. We identified microbes with robust inhibitory activity against L. crescens that include the fungi Cladosporium cladosporioides and Epicoccum nigrum and bacterial species of Pantoea, Bacillus, and Curtobacterium Purified bioactive natural products with anti-"Ca. Liberibacter asiaticus" activity were identified from the fungus C. cladosporioides Bioassay-guided fractionation of an organic extract of C. cladosporioides yielded the natural products cladosporols A, C, and D as the active agents against L. crescens This work serves as a foundation for unraveling the complex chemistries associated with the citrus microbiome to begin to understand the functional roles of members of the microbiome, with the long-term goal of developing anti-"Ca Liberibacter asiaticus" bioinoculants that thrive in the citrus holosystem.IMPORTANCE Globally, citrus is threatened by huanglongbing (HLB), and the lack of effective control measures is a major concern of farmers, markets, and consumers. There is compelling evidence that plant health is a function of the activities of the plant's associated microbiome. Using Liberibacter crescens, a culturable surrogate for the unculturable HLB-associated bacterium "Candidatus Liberibacter asiaticus," we tested the hypothesis that members of the citrus microbiome produce potential anti-"Ca Liberibacter asiaticus" natural products with potential anti-"Ca Liberibacter asiaticus" activity. A subset of isolates obtained from the microbiome inhibited L. crescens growth in an agar diffusion inhibition assay. Further fractionation experiments linked the inhibitory activity of the fungus Cladosporium cladosporioides to the fungus-produced natural products cladosporols A, C, and D, demonstrating dose-dependent antagonism to L. crescens

An Update on the Status of the NIF Power Conditioning System

The National Ignition Facility (NIF) Power Conditioning System provides the pulsed excitation required to drive flashlamps in the laser's optical amplifiers. Modular in design, each of the 192 Main Energy Storage Modules (MESMs) stores up to 2.2 MJ of electrical energy in its capacitor bank before delivering the energy to 20 pairs of flashlamps in a 400 {micro}s pulse (10% power points). The peak current of each MESM discharge is 0.5 MA. Production, installation, commissioning and operation of the NIF Power Conditioning continue to progress rapidly, with the goals of completing accelerated production and commissioning by early 2008, while maintaining an aggressive operation schedule. To date, more than 97% of the required modules have been assembled, shipped and installed in the facility, representing more that 380 MJ of stored energy available for driving NIF flashlamps. The MESMs have displayed outstanding reliability during daily, multiple-shift operations

Self-reported poor quality of sleep in solid organ transplant : A systematic review

Background High quality sleep of sufficient duration is vital to overall health and wellbeing. Self-reported poor quality of sleep, sleep reported as irregular in timing, marked by frequent awakenings, or shortened in duration, is common across the solid-organ transplant trajectory. Aim This Systematic Review aimed to summarize available literature on rates of self-reported poor quality of sleep among solid organ transplant candidates and recipients. Methods A systematic search of published literature was conducted in PubMed/MEDLINE, Embase, Web of Science, CINHAL, and PsychInfo databases with no date restrictions. Original articles in the English language describing self-reported quality of sleep using standardized questionnaires in adults either waitlisted for, or who received a solid organ transplant (heart, lung, kidney, liver, pancreas, or multi-solid organ) were included. Results Of a potential 2054 articles identified, 44 were included (63.6% renal transplant, 20.5% liver transplant, 11.4% lung transplant, and 4.5% included multiple organ transplant populations), with the majority (68.2%) focusing only on post-transplant populations. No included articles focused solely on heart or pancreas transplant populations. On average, the transplant population with the greatest improvement in quality of sleep (reported as poor sleep quality, insomnia, sleep disturbance, or sleep dissatisfaction) from transplant candidacy to post-transplantation were renal transplant (from 53.5% pre, to 38.9% post) followed by liver transplant patients (from 52.8% pre, to 46.3% post), while lung transplant patients remained similar pre- to post-transplantation (55.6% pre, to 52% post). Poor quality of sleep was frequently associated with anxiety and depression, poorer quality of life, restless legs syndrome, and higher comorbidity. Conclusions Reports of poor quality of sleep are highly prevalent across all solid-organ transplant populations, both pre- and post-transplantation. Future studies should assess quality of sleep longitudinally throughout all phases of the transplantation trajectory, with more research focusing on how to optimize sleep in solid organ transplant populations

Feelings of Emotional Exhaustion and Depersonalization Predict Self-Report of Trouble Sleeping for Nurses Working in Hospital Environments

Introduction American nurses report sleeping an average of 6.8 hours per night on workdays, which is less than the recommended 7-9 hours. Burnout, which is comprised of emotional exhaustion (EE), depersonalization (DP) and personal accomplishment (PA), is common among nurses and may contribute to insufficient sleep quantity and sleep disruption. We aimed to investigate if burnout, measured using the Maslach Burnout Inventory (MBI), predicted self-report of trouble sleeping for nurses working in the hospital. Methods This is a secondary report of a prospective crossover trial of nurses assigned to either 6 weeks of daily work-breaks in an outdoor hospital garden or 6 weeks of indoor-only breaks. After a 1-week washout period, break assignments were switched for an additional 6 weeks. Nurses completed the MBI at the beginning and end of each 6-week period. Each workday, nurses completed a visual analog scale reporting no trouble sleeping to much trouble sleeping immediately following a break. For this analysis, we used initial MBI subscale score (EE, DP, PA) as a predictor of mean trouble sleeping over the subsequent 6-week period regardless of break location using generalized estimating equations. Results A total of 29 nurses (27 females, mean age 42.8 years) participated. Most (n=20) worked dayshift, and 9 worked night shift. Initial MBI subscale score, regardless of break location, was predictive of mean self-report of trouble sleeping over the subsequent 6-week period for EE (p<0.001) and DP (p=0.004), but not for PA (p=0.48). When divided by break assignment (indoor-only vs garden), results were similar for EE (indoor p<0.001; garden p<0.001), DP (indoor p<0.001; garden p<0.001), and PA (indoor p=0.20; garden p=0.49). Conclusion The MBI subscales of EE and DP were predictive of self-report of trouble sleeping for nurses in this study. Further research using objective measures of sleep are needed to confirm and extend these preliminary observations. Support (If Any) Funding for this research was provided by the TKF Foundation as part of the National Open Spaces Sacred Places Initiative

ICU Patient Family Stress Recovery During Breaks in a Hospital Garden and Indoor Environments

Objectives: Measure the immediate change in intensive care unit (ICU) family members’ state stress levels from the beginning to the end of a person’s visit to a hospital garden and compare the changes produced by the garden with those associated with spending time in indoor hospital environments intended for respite and relaxation. Background: No previous research has compared the efficacy of different physical environments as interventions to foster stress reduction in family members of ICU patients, a group of hospital visitors known to experience high levels of distress. Method: A convenience sample of 42 ICU patient family (from 42 different families) completed the Present Functioning Visual Analogue Scales (PFVAS) before and after each visit (128 total visits) to a garden, an atrium/caf\ue9, or ICU waiting room. Results: Stress scores significantly declined (i.e., improved) from the start to the end of a break on all PFVAS subscales (p <.0001) in both the garden and indoors locations. However, it is noteworthy that garden breaks resulted in significantly greater improvement in the “sadness” scale than breaks in indoor locations (p =.03), and changes in all five other PFVAS scores showed somewhat more reduction of stress for breaks spent in the garden than indoors, although these differences were not statistically significant. Conclusion: Creating an unlocked garden with abundant nature located close to an ICU can be an effective intervention for significantly mitigating state stress in family members of ICU patients and can be somewhat more effective than indoor areas expressly designed for family respite and relaxation

Impact of nurses taking daily work breaks in a hospital garden on Burnout

Background Nurses working in hospital environments are at risk for burnout. Exposure to nature has psychological benefits, but the effect of hospital gardens on nurse burnout is less understood. Objective To compare the effect on nurse burnout of taking daily work breaks in a hospital-integrated garden with the effect of indoor-only breaks. Methods A prospective crossover trial was conducted of nurses assigned to either 6 weeks of a work break in an outdoor hospital garden or 6 weeks of indoor-only breaks. After a 1-week washout period, break assignments were switched for a subsequent 6 weeks. The Maslach Burnout Inventory was administered at the beginning and end of each 6-week period, and a Present Functioning Visual Analogue Scale was completed at the start and end of each break to capture immediate psychological symptoms. Change scores were analyzed by using generalized estimating equations. Results For 29 nurses, for garden compared with indoor breaks, significant improvement was apparent in scores on the Maslach Burnout Inventory subscales for emotional exhaustion (4.5 vs -0.2; P < .001) and depersonalization (1.8 vs 0.0; P = .02) but not for personal accomplishment (-0.6 vs -0.0; P = .55). Compared with indoor breaks, total symptom scores on the Present Functioning Visual Analog Scale improved significantly when nurses took a break in the garden (garden vs indoor breaks, 4.0 vs 2.4; P = .04). Conclusions Taking daily work breaks in an outdoor garden may be beneficial in mitigating burnout for nurses working in hospital environments

An In Vitro Pipeline for Screening and Selection of Citrus-Associated Microbiota with Potential Anti-"Candidatus Liberibacter asiaticus" Properties.

Huanglongbing (HLB) is a destructive citrus disease that is lethal to all commercial citrus plants, making it the most serious citrus disease and one of the most serious plant diseases. Because of the severity of HLB and the paucity of effective control measures, we structured this study to encompass the entirety of the citrus microbiome and the chemistries associated with that microbial community. We describe the spatial niche diversity of bacteria and fungi associated with citrus roots, stems, and leaves using traditional microbial culturing integrated with culture-independent methods. Using the culturable sector of the citrus microbiome, we created a microbial repository using a high-throughput bulk culturing and microbial identification pipeline. We integrated an in vitro agar diffusion inhibition bioassay into our culturing pipeline that queried the repository for antimicrobial activity against Liberibacter crescens, a culturable surrogate for the nonculturable "Candidatus Liberibacter asiaticus" bacterium associated with HLB. We identified microbes with robust inhibitory activity against L. crescens that include the fungi Cladosporium cladosporioides and Epicoccum nigrum and bacterial species of Pantoea, Bacillus, and Curtobacterium Purified bioactive natural products with anti-"Ca. Liberibacter asiaticus" activity were identified from the fungus C. cladosporioides Bioassay-guided fractionation of an organic extract of C. cladosporioides yielded the natural products cladosporols A, C, and D as the active agents against L. crescens This work serves as a foundation for unraveling the complex chemistries associated with the citrus microbiome to begin to understand the functional roles of members of the microbiome, with the long-term goal of developing anti-"Ca Liberibacter asiaticus" bioinoculants that thrive in the citrus holosystem.IMPORTANCE Globally, citrus is threatened by huanglongbing (HLB), and the lack of effective control measures is a major concern of farmers, markets, and consumers. There is compelling evidence that plant health is a function of the activities of the plant's associated microbiome. Using Liberibacter crescens, a culturable surrogate for the unculturable HLB-associated bacterium "Candidatus Liberibacter asiaticus," we tested the hypothesis that members of the citrus microbiome produce potential anti-"Ca Liberibacter asiaticus" natural products with potential anti-"Ca Liberibacter asiaticus" activity. A subset of isolates obtained from the microbiome inhibited L. crescens growth in an agar diffusion inhibition assay. Further fractionation experiments linked the inhibitory activity of the fungus Cladosporium cladosporioides to the fungus-produced natural products cladosporols A, C, and D, demonstrating dose-dependent antagonism to L. crescens

Biosynthesis of Guanitoxin Enables Global Environmental Detection in Freshwater Cyanobacteria

Harmful cyanobacterial blooms (cyanoHABs) cause recurrent toxic events in global watersheds. Although public health agencies monitor the causal toxins of most cyanoHABs and scientists in the field continue developing precise detection and prediction tools, the potent anticholinesterase neurotoxin, guanitoxin, is not presently environmentally monitored. This is largely due to its incompatibility with widely employed analytical methods and instability in the environment, despite guanitoxin being among the most lethal cyanotoxins. Here, we describe the guanitoxin biosynthesis gene cluster and its rigorously characterized nine-step metabolic pathway from l-arginine in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024. Through environmental sequencing data sets, guanitoxin (gnt) biosynthetic genes are repeatedly detected and expressed in municipal freshwater bodies that have undergone past toxic events. Knowledge of the genetic basis of guanitoxin biosynthesis now allows for environmental, biosynthetic gene monitoring to establish the global scope of this neurotoxic organophosphate

Biosynthesis of Guanitoxin Enables Global Environmental Detection in Freshwater Cyanobacteria

Harmful cyanobacterial blooms (cyanoHABs) cause recurrent toxic events in global watersheds. Although public health agencies monitor the causal toxins of most cyanoHABs and scientists in the field continue developing precise detection and prediction tools, the potent anticholinesterase neurotoxin, guanitoxin, is not presently environmentally monitored. This is largely due to its incompatibility with widely employed analytical methods and instability in the environment, despite guanitoxin being among the most lethal cyanotoxins. Here, we describe the guanitoxin biosynthesis gene cluster and its rigorously characterized nine-step metabolic pathway from l-arginine in the cyanobacterium Sphaerospermopsis torques-reginae ITEP-024. Through environmental sequencing data sets, guanitoxin (gnt) biosynthetic genes are repeatedly detected and expressed in municipal freshwater bodies that have undergone past toxic events. Knowledge of the genetic basis of guanitoxin biosynthesis now allows for environmental, biosynthetic gene monitoring to establish the global scope of this neurotoxic organophosphate