Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR.

Background. Antimicrobial resistance (AMR) is an ever-increasing global health concern. One crucial facet in tackling the AMR epidemic is earlier and more accurate AMR diagnosis, particularly in the dangerous and highly multi-drug-resistant ESKAPE pathogen, Pseudomonas aeruginosa.Objectives. We aimed to develop two SYBR Green-based mismatch amplification mutation assays (SYBR-MAMAs) targeting GyrA T83I (gyrA248) and GyrA D87N, D87Y and D87H (gyrA259). Together, these variants cause the majority of fluoroquinolone (FQ) AMR in P. aeruginosa.Methods. Following assay validation, the gyrA248 and gyrA259 SYBR-MAMAs were tested on 84 Australian clinical P. aeruginosa isolates, 46 of which demonstrated intermediate/full ciprofloxacin resistance according to antimicrobial susceptibility testing.Results. Our two SYBR-MAMAs correctly predicted an AMR phenotype in the majority (83%) of isolates with intermediate/full FQ resistance. All FQ-sensitive strains were predicted to have a sensitive phenotype. Whole-genome sequencing confirmed 100 % concordance with SYBR-MAMA genotypes.Conclusions. Our GyrA SYBR-MAMAs provide a rapid and cost-effective method for same-day identification of FQ AMR in P. aeruginosa. An additional SYBR-MAMA targeting the GyrB S466Y/S466F variants would increase FQ AMR prediction to 91 %. Clinical implementation of our assays will permit more timely treatment alterations in cases where decreased FQ susceptibility is identified, leading to improved patient outcomes and antimicrobial stewardship.

Express Yourself: Quantitative Real-Time PCR Assays for Rapid Chromosomal Antimicrobial Resistance Detection in Pseudomonas aeruginosa.

The rise of antimicrobial-resistant (AMR) bacteria is a global health emergency. One critical facet of tackling this epidemic is more rapid AMR diagnosis in serious multidrug-resistant pathogens like Pseudomonas aeruginosa. Here, we designed and then validated two multiplex quantitative real-time PCR (qPCR) assays to simultaneously detect differential expression of the resistance-nodulation-division efflux pumps MexAB-OprM, MexCD-OprJ, MexEF-OprN, and MexXY-OprM, the AmpC ß-lactamase, and the porin OprD, which are commonly associated with chromosomally encoded AMR. Next, qPCRs were tested on 15 sputa from 11 participants with P. aeruginosa respiratory infections to determine AMR profiles in vivo. We confirmed multiplex qPCR testing feasibility directly on sputa, representing a key advancement in in vivo AMR diagnosis. Notably, comparison of sputa with their derived isolates grown in Luria-Bertani broth (±2.5% NaCl) or a 5-antibiotic cocktail showed marked expression differences, illustrating the difficulty in replicating in vivo expression profiles in vitro. Cystic fibrosis sputa showed significantly reduced mexE and mexY expression compared with chronic obstructive pulmonary disease sputa, despite harboring fluoroquinolone- and aminoglycoside-resistant strains, indicating that these loci do not contribute to AMR in vivo. oprD was also significantly downregulated in cystic fibrosis sputa, even in the absence of contemporaneous carbapenem use, suggesting a common adaptive trait in chronic infections that may affect carbapenem efficacy. Sputum ampC expression was highest in participants receiving carbapenems (6.7 to 15×), some of whom were simultaneously receiving cephalosporins, the latter of which would be rendered ineffective by the upregulated ampC. Our qPCR assays provide valuable insights into the P. aeruginosa resistome, and their use on clinical specimens will permit timely treatment alterations that will improve patient outcomes and antimicrobial stewardship measures.

Genomic diversity and antimicrobial resistance of Prevotella species isolated from chronic lung disease airways.

Cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) are characterized by increasingly frequent acute pulmonary exacerbations that reduce life quality and length. Human airways are home to a rich polymicrobial environment, which includes members of the obligately anaerobic genus Prevotella. Despite their commonness, surprisingly little is known about the prevalence, role, genomic diversity and antimicrobial resistance (AMR) potential of Prevotella species and strains in healthy and diseased airways. Here, we used comparative genomics to develop a real-time PCR assay to permit rapid Prevotella species identification and quantification from cultures and clinical specimens. Assay specificity was validated across a panel of Prevotella and non-Prevotella species, followed by PCR screening of CF and COPD respiratory-derived cultures. Next, 35 PCR-positive isolates were subjected to whole-genome sequencing. Of eight identified Prevotella species, P. histicola, P. melaninogenica, P. nanceiensis, P. salivae and P. denticola overlapped between participant cohorts. Phylogenomic analysis revealed considerable interhost but limited intrahost diversity, suggesting patient-specific lineages in the lower airways, probably from oral cavity aspirations. Correlation of phenotypic AMR profiles with AMR genes identified excellent correlation between tetQ presence and decreased doxycycline susceptibility, and ermF presence and decreased azithromycin susceptibility and clindamycin resistance. AMR rates were higher in the CF isolates, reflecting greater antibiotic use in this cohort. All tested Prevotella isolates were tobramycin-resistant, providing a potential selection method to improve Prevotella culture retrieval rates. Our addition of 35 airway-derived Prevotella genomes to public databases will enhance ongoing efforts to unravel the role of this diverse and enigmatic genus in both diseased and healthy lungs.

Respiratory acute discharge service: a hospital in the home programme for chronic obstructive pulmonary disease exacerbations (RADS study).

BACKGROUND: Respiratory Acute Discharge Service (RADS) is a novel early discharge service with nurse-led community based recovery in selected patients with acute exacerbations of chronic obstructive pulmonary disease. AIM: This pilot study aimed to determine the efficacy and safety of the programme in an Australian tertiary hospital. METHODS: All patients who were recruited to RADS at Sunshine Coast University Hospital over a 6 months period from June to November 2018 were included. The co-primary outcomes were length of hospital days saved and rate of readmission within 30 days from discharge. RESULTS: A total of 166 patients (median age 74 years (interquartile range 70-80 years)) was recruited to the programme over the study period. The mean forced expiratory volume in one second (FEV1%) of the patients was 42% (standard deviation 19). The median length-of-stay prior to discharge on the RADS programme was 1 day (range 0-5), compared to a previous average of 5.8 days in our health service. Patients were on the programme for a median of 4 days (range 1-6). A total of 613 hospital bed days was saved over the study period, with significant cost savings. Forty-one (24.7%) patients represented to hospital within 30 days, the majority (64%) were due to recurrent symptoms. The rate of 30-day all-cause mortality for the study population was 1 (0.6%). CONCLUSION: Early supported discharge care model with nurse-led community based recovery after an acute exacerbation of chronic obstructive pulmonary disease in selected patients is safe, and has the potential to provide greater flow through the hospital systems with cost effective care.

A crossover study of short burst oxygen therapy (SBOT) for the relief of exercise-induced breathlessness in severe COPD.

BACKGROUND: Previous small studies suggested SBOT may be ineffective in relieving breathlessness after exercise in COPD. METHODS: 34 COPD patients with FEV1 <40% predicted and resting oxygen saturation ≥93% undertook an exercise step test 4 times. After exercise, patients were given 4 l/min of oxygen from a simple face mask, 4 l/min air from a face mask (single blind), air from a fan or no intervention. RESULTS: Average oxygen saturation fell from 95.0% to 91.3% after exercise. The mean time to subjective recovery was 3.3 minutes with no difference between treatments. The mean Borg breathlessness score was 1.5/10 at rest, rising to 5.1/10 at the end of exercise (No breathlessness = 0, worst possible breathlessness = 10). Oxygen therapy had no discernable effect on Borg scores even for 14 patients who desaturated below 90%. 15 patients had no preferred treatment, 7 preferred oxygen, 6 preferred the fan, 3 preferred air via a mask and 3 preferred room air. CONCLUSIONS: This study provides no support for the idea that COPD patients who are not hypoxaemic at rest derive noticeable benefit from oxygen therapy after exercise. Use of air from a mask or from a fan had no apparent physiological or placebo effect.

Nursing assessment of obstructive sleep apnea in hospitalised adults: a review of risk factors and screening tools.

Obstructive sleep apnea (OSA) affects approximately 2-4% of the general population and may be more prevalent in obese adults. However, sleep apnea remains consistently under-diagnosed in the general population as well as in hospital wards. Nurse awareness of OSA during routine monitoring could allow specific observations of hospitalised adults to identify those at high risk and ensure appropriate referral. This integrative literature review analysed major risk factors for OSA and identified screening tools that nurses could utilise in hospital wards. The most important risk factors relevant to nursing practice in hospital settings were obesity, hypertension and sleep position. The most suitable screening tool was the Berlin Questionnaire, while there was some evidence to support measuring waist circumference. A nursing assessment flow chart was developed based on the literature reviewed. This paper highlights a role for nurses in recognising patients at risk of OSA and minimising complications in hospitalised adults.

Quality nephrology nursing care: beyond Kt/V.

Through a critical review of nursing and medical literature, this article argues that nephrology nurses have embraced Kt/V at the expense of other core elements of nephrology nursing care. The focus on quality care as technical expertise may dominate at the expense of interpersonal care. Nurses need to challenge the influence Kt/V has on other aspects of nephrology nursing care.

Effectiveness of non-pharmacological interventions for fatigue in adults with multiple sclerosis, rheumatoid arthritis, or systemic lupus erythematosus: a systematic review.

AIM: This paper reports a systematic review of non-pharmacological interventions for fatigue in adults with three common autoimmune conditions. BACKGROUND: A considerable proportion of people with multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus experience compromised quality of life due to fatigue. Recent reviews of pharmacotherapies for fatigue in these conditions remain inconclusive, and systematic evidence for effectiveness of non-pharmacological interventions was unavailable. Our paper addresses this gap. METHODS: The literature search used the key words fatigue, energy, multiple sclerosis, rheumatoid arthritis and systemic lupus. It included 19 electronic databases and libraries, three evidence-based journals, two internet search engines, was dated 1987-2006, and limited to English. Non-pharmacological experimental studies about fatigue comprising more than five adults were included. Meta-analysis was not possible due to diverse interventions and outcome measures, therefore studies were analysed by types of interventions used to reduce fatigue. RESULTS: Of 653 hits, 162 papers were reviewed, and 36 met the inclusion criteria. Thirty-three primary studies reported 14 randomized controlled trials and 19 quasi-experimental designs. Most interventions were tested with people with multiple sclerosis. Exercise, behavioural, nutritional and physiological interventions were associated with statistically significant reductions in fatigue. Aerobic exercise was effective, appropriate and feasible for reducing fatigue among adults with chronic autoimmune conditions. Electromagnetic field devices showed promise. The diversity of interventions, designs, and using 24 different instruments to measure fatigue, limited comparisons. CONCLUSION: Low impact aerobic exercise gradually increasing in intensity, duration and frequency may be an effective strategy in reducing fatigue in some adults with chronic auto-immune conditions. However, fatigue is a variable and personal experience and a range of behavioural interventions may be required. Well-designed studies testing these promising strategies and consensus on outcome fatigue measures are needed.

Evidence for early oral feeding of patients after elective open colorectal surgery: a literature review.

AIM: To review research on early oral feeding following elective, open colorectal surgery. BACKGROUND: Fasting following gastrointestinal surgery is a traditional surgical practice, based on fears of causing postoperative complications if oral intake begins before bowel function returns, but fasting following elective surgery is questionable as a best practice. METHODS: Searches in Journals@Ovid CINAHL, MEDLINE, PubMed, Web of Science and The Cochrane Library for primary studies, published during 1995-2004, used the keywords: 'surgery', 'postoperative', 'elective, 'colorectal', 'bowel, 'colon', 'oral', 'enteral', 'feeding', 'early', 'traditional'. Studies of adults undergoing elective, open colorectal surgery who were allowed fluids and food before bowel function returned (early feeding) were included. Outcomes of interest were safety, tolerability, duration of gastrointestinal ileus and length of hospital stay. Critical appraisal of randomized and controlled studies was undertaken following inclusion. RESULTS: Fifteen studies comprising 1352 patients were reviewed. All studies concluded early feeding was safe, based on complications rates. Total complications were 12.5% (range 0-25%) for 935 early feeding patients, with no increased risk of anastomotic leak, aspiration pneumonia, or bowel obstruction. For all studies an average of 86% patients (range 73-100%) tolerated early feeding. Studies demonstrating faster resolution of postoperative ileus or shorter hospitalization were associated with multimodal perioperative care, including early mobilization, epidural analgesia and comprehensive patient education. Appraisal of five randomized trials revealed no blinding and inadequate randomization. CONCLUSIONS: This review supports early oral feeding after elective, open colorectal surgery and challenges the traditional practice of fasting patients until return of bowel function. Early feeding was safe, well-tolerated and easy to implement. Reduced length of ileus and shorter hospitalization may occur with multimodal protocols. RELEVANCE TO CLINICAL PRACTICE: Nurses can highlight this new evidence for other health professionals, advocate development of clinical protocols featuring early feeding and participate in multi-disciplinary, multi-method research regarding benefits of early feeding.

Health as expanding consciousness: seven women living with multiple sclerosis or rheumatoid arthritis.

The meaning of health as expanding consciousness is explored through stories of seven women who developed multiple sclerosis or rheumatoid arthritis during their lives. Using Newman's hermeneutic-dialectic approach, unstructured interviews were conducted over a 2-year period. Analysis and interpretation of narratives concerning person-environment interactions revealed turning points and separate choice points before four new ways of living including finding simple pleasures, being positive, gaining self-control, and self-differentiation, were found. Support for Newman's stages of expanding consciousness and more comprehensive descriptions of self-transcendence in space and time are presented. Implications for theory development and theory-guided practice are offered.

Exploring underlying life patterns of women with multiple sclerosis or rheumatoid arthritis: comparison with NANDA dimensions.

In Newman's theory, disease is one of many manifestations of underlying pattern and its existence provides meaningful information about person-environment interactions. Underlying patterns manifest differently over time, so clues to their understanding can be found within life stories. Further interpretation subsequent to illustrating expanding consciousness for seven women living with multiple sclerosis or rheumatoid arthritis suggested six underlying patterns expressed in theoretical terms as energy-fatigue, giving-receiving, rejecting-accepting, vulnerability-resilience, control-release, and being silent-speaking out. Discussion and comparison with the North American Nursing Diagnosis Association's dimensions for assessment of human response patterns illustrates how nurses caring for women could identify and use underlying patterns in practice.

The Academy at Harvard Medical School: nurturing teaching and stimulating innovation.

The Academy at Harvard Medical School, established in 2001, was formed at a critical moment for medical schools in this country. Several decades of enormous growth in the biomedical research and clinical care activities of medical school faculty have resulted in great societal benefit. The unintended consequence has been a decline in faculty time and reward for the educational mission that is unique to a medical school. The impact of this decline is particularly felt now because the explosive growth in the science and technology relevant to medical practice, coupled with dramatic changes in the health care delivery system, calls for new models for the education of the next generation of physicians. The mission of the academy is to renew and reinvigorate the educational mission of Harvard Medical School (HMS). By bringing together a select group of some of the school's most talented and dedicated faculty and providing direct support for their work related to education, the academy has created a unique mechanism for increasing the recognition of teaching contributions of both academy members and the teaching faculty at large, fostering educational innovation, and providing a forum for the exchange of ideas related to medical education that cross departmental and institutional lines. The authors describe the academy's membership criteria, structure, governance, activities, institutional impact, and plans for long-term evaluation, and indicate challenges the academy will face in the future.

Genome scan of pedigrees multiply affected with bipolar disorder provides further support for the presence of a susceptibility locus on chromosome 12q23-q24, and suggests the presence of additional loci on 1p and 1q.

OBJECTIVE: To localize genes conferring susceptibility to bipolar affective disorder. METHODS: Seven families were selected on the basis of containing multiple cases of bipolar affective disorder present in three or more generations, an absence of schizophrenia and unilineal transmission. DNA samples from these families were genotyped with 365 microsatellite markers spaced at approximately 10 cM intervals across the whole genome. All markers were subjected to initial two-point and three-point analyses using LOD score and model-free analysis. All regions producing a result significant at P<0.01 were then subjected to four-point LOD score analysis under the assumption of heterogeneity. RESULTSA four-point LOD score of 2.8 was obtained using a dominant model and including unipolar cases as affected in the region of D12S342. Four-point LOD scores of 2 were obtained around D1S243, D1S251 and D3S1265. The positive results around D1S243 were accounted for by a LOD score of 3.1 occurring in a single pedigree. CONCLUSIONS: Since there has been previous strong support for linkage to the region of 12q23-q24 around D12S342, it now seems very probable that it does indeed contain a gene influencing susceptibility to bipolar affective disorder. Some evidence for linkage in the region of 1q near to D1S251 has been reported in one previous study. It therefore seems that this region of 1q and the region of 1p close to D1S243 may also harbour susceptibility genes.

Undergraduate nursing students' clinical experiences in rural and remote areas: recruitment implications.

Providing rural and remote clinical experiences for undergraduate nursing students could help address future recruitment in these areas, but adequate financial support for students during clinical placements is also necessary. Strategies to address nursing shortages to date have emphasised funding for rural re-entry programs, or supporting students from rural backgrounds to attend university courses. Assisting current undergraduate nursing students, especially those from urban backgrounds, to experience living and working in rural areas has been overlooked as a potential recruitment strategy. Ongoing qualitative evaluation of a clinical placement program shows students respond positively and with increased interest in returning to rural nursing after graduation. Unfortunately, many are disadvantaged financially by the added expense of their rural clinical rotation. Finding ways to support students from both urban and non-urban backgrounds to undertake rural and remote clinical placements should be an important strategic and funding priority.

Transcendence and transformation in the life patterns of women living with rheumatoid arthritis.

Considering personal life stories as the context for health transitions can enhance understanding of what is meaningful in living with chronic illness. Informed by Margaret Newman's theory of Health as Expanding Consciousness, this interpretive study described the life patterns of three women with rheumatoid arthritis as a process of expanding consciousness. The women's stories revealed transcendence of self-boundaries and personal transformation as new ways of living, including "simple pleasures" and "being positive." Through understanding life patterns within caring nursing partnerships, transitions in an entire life story can be appreciated as complex processes involving transcendence and transformation.