Chronic infection phenotypes of Pseudomonas aeruginosa are associated with failure of eradication in children with cystic fibrosis.

Early eradication treatment with inhaled tobramycin is successful in the majority of children with cystic fibrosis (CF) with incident Pseudomonas aeruginosa infection. However, in 10-40 % of cases, eradication fails and the reasons for this are poorly understood. The purpose of this study was to determine whether specific microbial characteristics could explain eradication treatment failure. This was a cross-sectional study of CF patients (aged 0-18 years) with incident P. aeruginosa infection from 2011 to 2014 at the Hospital for Sick Children, Toronto, Canada. Phenotypic assays were done on all incident P. aeruginosa isolates, and eradicated and persistent isolates were compared using the Mann-Whitney test or the two-sided Chi-square test. A total of 46 children with CF had 51 incident P. aeruginosa infections. In 72 % (33/46) of the patients, eradication treatment was successful, while 28 % failed eradication therapy. Persistent isolates were less likely to be motile, with significantly less twitch motility (p=0.001), were more likely to be mucoid (p=0.002), and more likely to have a tobramycin minimum inhibitory concentration (MIC) ≥ 128 µg/mL (p=0.02) compared to eradicated isolates. Although biofilm production was similar, there was a trend towards more persistent isolates with deletions in quorum-sensing genes compared with eradicated isolates (p=0.06). Initial acquisition of P. aeruginosa with characteristics of chronic infection is associated with failure of eradication treatment.

A non-destructive genotyping system from a single seed for marker-assisted selection in watermelon.

Genomic tools for watermelon breeding are becoming increasingly available. A high throughput genotyping system would facilitate the use of DNA markers in marker-assisted selection. DNA extraction from leaf material requires prior seed germination and is often time-consuming and cost prohibitive. In an effort to develop a more efficient system, watermelon seeds of several genotypes and various seed sizes were sampled by removing ⅓ or ½ sections from the distal ends for DNA extraction, while germinating the remaining proximal parts of the seed. Removing ⅓ of the seed from the distal end had no effect on seed germination percentage or seedling vigor. Different DNA extraction protocols were tested to identify a method that could yield DNA of sufficient quality for amplification by polymerase chain reaction. A sodium dodecyl sulfate extraction protocol with 1% polyvinylpyrrolidone yielded DNA that could be amplified with microsatellite primers and was free of pericarp contamination. In this study, an efficient, non-destructive genotyping protocol for watermelon seed was developed.

Conjugation between bacterial and mammalian cells.

Bacterial conjugation, in which DNA is transferred from one bacterium to another, was first reported in 1946 and found to be mediated by the F factor. Although the F and RK2/RP4 prototypic plasmids can mediate the transfer of DNA from bacteria to yeast, there has been no evidence of classical bacterial conjugation to higher eukaryotes. Here, I present evidence of such transfer, using Escherichia coli, the RK2 plasmid system and Chinese hamster ovary CHO K1 cells.

Comparison of three typing methods for Pseudomonas aeruginosa isolates from patients with cystic fibrosis.

The aim of this study was to compare two traditional pattern matching techniques, pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD), with the more reproducible technique of multilocus sequence typing (MLST) to genotype a blinded sample of Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients. A blinded sample of 48 well-characterized CF P. aeruginosa isolates was genotyped by PFGE, RAPD, and MLST, each performed in a different laboratory. The discriminatory power and congruence between the methods were compared using the Simpson's index, Rand index, and Wallace coefficient. PFGE and MLST had the greatest congruence with the highest Rand index (0.697). The discriminatory power of PFGE, RAPD, and MLST were comparable, with high Simpson's indices (range 0.973-0.980). MLST identified the most clonal relationships. When clonality was defined as agreement between two or more methods, MLST had the greatest predictive value (100 %) in labeling strains as unique, while PFGE had the greatest predictive value (96 %) in labeling strains as clonal. This study demonstrated the highest level of agreement between PFGE and MLST in genotyping P. aeruginosa isolates from CF patients. MLST had the greatest predictive value in identifying strains as unique and, thus, has the potential to be a cost-efficient, high-throughput, first-pass typing method.

The culture, mental health and psychosocial wellbeing of Rohingya refugees: a systematic review.

AIMS: Despite the magnitude and protracted nature of the Rohingya refugee situation, there is limited information on the culture, mental health and psychosocial wellbeing of this group. This paper, drawing on a report commissioned by the United Nations High Commissioner for Refugees (UNHCR), aims to provide a comprehensive synthesis of the literature on mental health and psychosocial wellbeing of Rohingya refugees, including an examination of associated cultural factors. The ultimate objective is to assist humanitarian actors and agencies in providing culturally relevant Mental Health and Psychosocial Support (MHPSS) for Rohingya refugees displaced to Bangladesh and other neighbouring countries. METHODS: We conducted a systematic search across multiple sources of information with reference to the contextual, social, economic, cultural, mental health and health-related factors amongst Rohingya refugees living in the Asia-Pacific and other regions. The search covered online databases of diverse disciplines (e.g. medicine, psychology, anthropology), grey literature, as well as unpublished reports from non-profit organisations and United Nations agencies published until 2018. RESULTS: The legacy of prolonged exposure to conflict and persecution compounded by protracted conditions of deprivations and displacement is likely to increase the refugees' vulnerability to wide array of mental health problems including posttraumatic stress disorder, anxiety, depression and suicidal ideation. High rates of sexual and gender-based violence, lack of privacy and safe spaces and limited access to integrated psychosocial and mental health support remain issues of concern within the emergency operation in Bangladesh. Another challenge is the limited understanding amongst the MHPSS personnel in Bangladesh and elsewhere of the language, culture and help-seeking behaviour of Rohingya refugees. While the Rohingya language has a considerable vocabulary for emotional and behavioural problems, there is limited correspondence between these Rohingya terms and western concepts of mental disorders. This hampers the provision of culturally sensitive and contextually relevant MHPSS services to these refugees. CONCLUSIONS: The knowledge about the culture, context, migration history, idioms of distress, help-seeking behaviour and traditional healing methods, obtained from diverse sources can be applied in the design and delivery of culturally appropriate interventions. Attention to past exposure to traumatic events and losses need to be paired with attention for ongoing stressors and issues related to worries about the future. It is important to design MHPSS interventions in ways that mobilise the individual and collective strengths of Rohingya refugees and build on their resilience.

Staphylococcus aureus interaction with Pseudomonas aeruginosa biofilm enhances tobramycin resistance.

Antimicrobial resistance is a significant threat to the treatment of infectious disease. Multiple mechanisms of resistance to different classes of antibiotics have been identified and well-studied. However, these mechanisms are studied with bacteria in isolation, whereas often, infections have a polymicrobial basis. Using a biofilm slide chamber model, we visualized the formation and development of clinical Pseudomonas aeruginosa biofilms in the presence of secreted Staphylococcus aureus exoproducts, two bacteria that commonly co-infect pediatric patients with cystic fibrosis. We showed that, over time, certain isolates of P. aeruginosa can form different biofilm architecture in the presence of S. aureus exoproducts. We further determined that this interaction was dependent on Psl produced by P. aeruginosa and staphylococcal protein A from S. aureus. Importantly, we identified a mechanism of antibiotic resistance to tobramycin that is dependent on the polymicrobial interactions between these two bacteria. This interaction occurred in isolates of P. aeruginosa recovered from children with cystic fibrosis who failed to clear P. aeruginosa following inhaled tobramycin treatment.

The dissociation of renin and aldosterone during critical illness.

A syndrome of elevated PRA accompanied by inappropriately low plasma aldosterone (ALDO) levels has been identified in some critically ill patients. To determine whether this phenomenon is due to a disturbance in factors that stimulate ALDO, we measured PRA, angiotensin II (AII), potassium (K+), and ACTH levels in 83 patients admitted to an intensive care unit. In 59 patients, PRA was greater than 2.0 ng/ml X h. Of these, 24 had an ALDO to PRA ratio (ALDO/PRA) below 2 (group I), and 35 had an ALDO/PRA ratio of 2 or more (group II). An ALDO/PRA ratio below 2 was deemed inappropriately low. Despite markedly elevated PRA [34 +/- 12 (+/- SE) ng/ml X h], the group I patients had inappropriately low ALDO levels (19 +/- 5 ng/dL). Patients in group II had significantly higher ALDO levels (48 +/- 6 ng/dL) despite lower PRA (9 +/- 1 ng/ml X h). AII levels were appropriately elevated in group I (39 +/- 26 pg/mL) and significantly greater (P less than 0.5) than those in group II. PRA correlated well with AII in both groups. There were no differences in plasma ACTH or K+ in these 2 groups, and plasma cortisol levels were similarly elevated in both groups of patients. Of 66 consecutively studied patients, 14 (21%) had inappropriate ALDO (group I). Mortality was significantly greater in group I (75%) than in group II (46%; P less than 0.001). In summary, a significant subset (21%) of seriously ill patients have inappropriately low ALDO levels despite elevated PRA. This dissociation is not due to an impairment of AII production or changes in plasma ACTH or K+. This phenomenon is associated with a higher mortality during critical illness. In light of evidence of decreased adrenal androgen secretion during severe illness, this dissociation of renin and aldosterone may represent an additional adrenal adaptation designed to promote cortisol production in critically ill patients.

Conjugative transfer in the dissemination of beta-lactam and aminoglycoside resistance.

The dissemination of antibiotic resistance among pathogenic bacteria can be attributed largely to conjugative DNA transfer. The general category of conjugative transfer includes both bacterial plasmid conjugation and the transfer of nonreplicative conjugative transposons. Prototypes for these two systems are the plasmid RK2 and the conjugative transposon Tn916. To address the long-term problem of the increasing prevalence and severity of antibiotic resistance, strategies aimed against conjugative transfer are needed, but their development will require a greater understanding of conjugative resistance gene acquisition. Overviews of the two conjugative transfer systems are presented, to summarize and compare current concepts. Observations regarding transfer of conjugative transposons is consistent with the prevailing model for plasmid conjugation, that is, by the transfer of a single-stranded DNA molecule from the donor to the recipient bacterium, and the generation of the single strand by rolling circle DNA replication. The relevance of vegetative plasmid replication and host range to the spread of multiple drug resistance is discussed, and clinical examples of conjugative transfer of multiple antibiotic resistance illustrate the severity of the current situation. Possible directions, traditional and innovative, are offered to address the conjugative transfer problem in drug resistance, and potentially to break the cycle of antibiotic development followed by the bacterial resistance gene acquisition.

Etiology of community-acquired pediatric viral diarrhea: a prospective longitudinal study in hospitals, emergency departments, pediatric practices and child care centers during the winter rotavirus outbreak, 1997 to 1998. The Pediatric Rotavirus Epidemiology Study for Immunization Study Group.

OBJECTIVE: To determine the viral etiology of community-acquired diarrhea in children admitted to hospitals and presenting in emergency departments, pediatric practices and child care centers from November 1, 1997, to June 30, 1998. STUDY DESIGN: Children with diarrhea were identified in a prospective multisite cohort study and analyzed according to age, gender and duration of hospitalization. Stools were tested for rotavirus by enzyme immunoassay and for all other enteric viruses by electron microscopy. RESULTS: Of the 2524 children identified with diarrhea, stools of 1386 (55%) were tested by enzyme immunoassay for rotavirus, and of these 1365 (54%) were screened by electron microscopy for all identifiable enteric viruses. Rotavirus was found in 32% (n = 437), adenovirus in 4% (n = 55), torovirus in 3% (n = 44), Norwalk-like viruses in 2% (n = 25) and astrovirus (n = 14) and calicivirus (n = 7) in fewer than 1% of the specimens tested. The proportion of rotavirus was significantly higher in children 12 to 23 months of age (43% of tested stools, n = 159) and 24 to 35 months of age (38% of tested stools, n = 64) (P < 0.001) than in any other age group. Toroviruses were found to approximately the same extent in children > or =36 months of age (6% of tested stools, n = 19) as those <36 months of age. Rotavirus (36% of tested stools, n = 375, P < 0.0005) and torovirus (4% of tested stools, n = 43, P < 0.004) were most often found in hospitalized patients. In contrast Norwalk-like viruses (P < 0.001) and astroviruses (P < 0.01) were more commonly detected in specimens from patients who presented to physicians' offices and who were symptomatic for gastroenteritis in child care centers. CONCLUSION: This study demonstrates that although all known gastroenteritis viruses were diagnosed in symptomatic children, rotavirus was the etiologic agent in most cases of diarrhea managed in the community and in the hospital.

A natural mutant of plasmid RP4 that confers phage resistance and reduced conjugative transfer.

A natural isolate of RP4 (PRC#116) acquired from the Stanford University Plasmid Reference Center differed from the wild-type Incompatibility Group P plasmid in several respects. Cells of Escherichia coli harboring PRC#116 were resistant to the IncP pili-specific bacteriophage PRD1 and GU5, and transferred this plasmid at a lower efficiency than the wild-type RP4. Phage sensitivity was restored, and transfer considerably improved in PRC#116+ bacteria transformed with plasmid constructs containing the origin of transfer (oriT region) of RP4. Mutant RP4 plasmids equivalent to PRC#116 were selected at a high frequency from an RP4+ E. coli population infected with PRD1 indicating that this RP4 variant may be the product of a very common mutation of the wild-type plasmid.

Community palliative care clinical nurse specialists' descriptions of stress in their work.

This study analysed the lived experiences of work stress of 21 community palliative care clinical nurse specialists from four centres in urban Essex, UK. Data were collected using an open-ended questionnaire and semi-structured interviews. Three major themes emerged: pressure of workload; relationships with health professionals; and the impact of the sadness of the client group. The pressure of workload was associated with nine sub-themes including staff shortages, client's or professionals' misperceptions of role, changing role, and erosion of normal support systems at work or at home. Relationships with some general practitioners seemed to cause most difficulty and social services seemed to have different priorities and to respond slowly. The impact of the sadness of the client group was only seen in four distinct situations. An overwork stress spiral is described that provides a salutary warning for managers, but also offers interesting solutions.

Essential factors in a community college-nursing home partnership.

The community college-nursing home partnership has two major objectives: to develop nursing potential in long-term care settings and to influence the redirection of associate degree nursing education to include active participation in long-term care settings. Successful partnerships are built on a commitment to high quality education for students and high quality service for patients. Other essential factors to a successful partnership include collaborative planning, pragmatic goal setting, clear communication, and mutual respect.

Cultivate corporate culture and diversity.

Elements of teamwork; commitment; and multiple, diverse workforces comprise a healthy organization, evident by the behaviors and attitudes of its employees.

Women's perceptions of caesarean birth: a Roy international study.

The purpose of this Roy adaptation model-based multi-site international mixed method study was to examine the relations of type of caesarean birth (unplanned/planned), number of caesarean births (primary/repeat), and preparation for caesarean birth to women's perceptions of and responses to caesarean birth. The sample included 488 women from the United States (n = 253), Finland (n = 213), and Australia (n = 22). Path analysis revealed direct effects for type of and preparation for caesarean birth on responses to caesarean birth, and an indirect effect for preparation on responses to caesarean birth through perception of birth the experience.