Dissolvable alginate hydrogel-based biofilm microreactors for antibiotic susceptibility assays.

Biofilms are found in many infections in the forms of surface-adhering aggregates on medical devices, small clumps in tissues, or even in synovial fluid. Although antibiotic resistance genes are studied and monitored in the clinic, the structural and phenotypic changes that take place in biofilms can also lead to significant changes in how bacteria respond to antibiotics. Therefore, it is important to better understand the relationship between biofilm phenotypes and resistance and develop approaches that are compatible with clinical testing. Current methods for studying antimicrobial susceptibility are mostly planktonic or planar biofilm reactors. In this work, we develop a new type of biofilm reactor-three-dimensional (3D) microreactors-to recreate biofilms in a microenvironment that better mimics those in vivo where bacteria tend to form surface-independent biofilms in living tissues. The microreactors are formed on microplates, treated with antibiotics of 1000 times of the corresponding minimal inhibitory concentrations (1000 × MIC), and monitored spectroscopically with a microplate reader in a high-throughput manner. The hydrogels are dissolvable on demand without the need for manual scraping, thus enabling measurements of phenotypic changes. Bacteria inside the biofilm microreactors are found to survive exposure to 1000 × MIC of antibiotics, and subsequent comparison with plating results reveals no antibiotic resistance-associated phenotypes. The presented microreactor offers an attractive platform to study the tolerance and antibiotic resistance of surface-independent biofilms such as those found in tissues.

Isolations of the Spring Viremia of Carp Virus in the Upper Mississippi River (USA), Including a New Host, the Quillback.

In July of 2018 and 2019, wild fish health surveys were conducted along the Wisconsin and Minnesota portions of the upper Mississippi River. Spring viremia of carp virus (SVCV) was isolated from Common Carp Cyprinus carpio as well as a newly identified host species, the Quillback Carpiodes cyprinus. Sanger sequencing of the gene encoding for the G protein revealed a high similarity of the Quillback isolate to various SVCV isolates identified from Common Carp that were collected during earlier wild fish health surveys and mortality events in the USA. Despite annual monitoring, this virus has been infrequently identified. The speculative role of native fish and invertebrates in allowing the virus to persist for long periods without detection is discussed.

Development of duplex qPCR targeting Carnobacterium maltaromaticum and Vagococcus salmoninarum.

In November 2018, Vagococcus salmoninarum was identified as the causative agent of a chronic coldwater streptococcosis epizootic in broodstock brook trout (Salvelinus fontinalis) at the Iron River National Fish Hatchery in Wisconsin, USA. By February 2019, the epizootic spread to adjacent raceways containing broodstock lake trout (Salvelinus namaycush), whereby fish were found to be coinfected with Carnobacterium maltaromaticum and V. salmoninarum. To differentiate these two pathogens and determine the primary cause of the lake trout morbidity, a quantitative real-time PCR (qPCR) was developed targeting the C. maltaromaticum phenylalanyl-tRNA synthase alpha subunit (pheS) gene. The qPCR was combined with a V. salmoninarum qPCR, creating a duplex qPCR assay that simultaneously quantitates C. maltaromaticum and V. salmoninarum concentrations in individual lake trout tissues, and screens presumptive isolates from hatchery inspections and wild fish from national fish hatchery source waters throughout the Great Lakes basin. Vagococcus salmoninarum and C. maltaromaticum were co-detected in broodstock brook trout from two tribal hatcheries and C. maltaromaticum was present in wild fish in source waters of several national fish hatcheries. This study provides a powerful new tool to differentiate and diagnose two emerging Gram-positive bacterial pathogens.

Dynamic Adaptive Response of Pseudomonas aeruginosa to Clindamycin/Rifampicin-Impregnated Catheters.

Pseudomonas aeruginosa is the most common Gram-negative pathogen causing nosocomial multidrug resistant infections. It is a good biofilm producer and has the potential for contaminating medical devices. Despite the widespread use of antibacterial-impregnated catheters, little is known about the impacts of antibacterial coating on the pathogenesis of P. aeruginosa. In this study, we investigated the adaptive resistance potential of P. aeruginosa strain PAO1 in response to continuous antibiotic exposure from clindamycin/rifampicin-impregnated catheters (CR-IC). During exposure for 144 h to clindamycin and rifampicin released from CR-IC, strain PAO1 formed biofilms featuring elongated and swollen cells. There were 545 and 372 differentially expressed proteins (DEPs) identified in the planktonic and biofilm cells, respectively, by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Both Cluster of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that the planktonic cells responded to the released antibiotics more actively than the biofilm cells, with metabolism and ribosomal biosynthesis-associated proteins being significantly over-expressed. Exposure to CR-IC increased the invasion capability of P. aeruginosa for Hela cells and upregulated the expression of certain groups of virulence proteins in both planktonic and biofilm cells, including the outer membrane associated (flagella, type IV pili and type III secretion system) and extracellular (pyoverdine) virulence proteins. Continuous exposure of P. aeruginosa to CR-IC also induced the overexpression of antibiotic resistance proteins, including porins, efflux pumps, translation and transcription proteins. However, these upregulations did not change phenotypic minimum inhibitory concentration (MIC) during the experimental timeframe. The concerning association between CR-IC and overexpression of virulence factors in P. aeruginosa suggests the need for additional investigation to determine if it results in adverse clinical outcomes.

Childhood Loss Experiences, Religiousness/Spirituality, and Hope as Predictors of Adult Life Satisfaction.

Purpose The purpose of this study was to explore how adaptive patterns of religiousness/spirituality and hope predict adult life satisfaction in adults, even if they had childhood loss experiences.Design and Methods Using a cross-sectional survey design with132 adult participants (mean age 33.8 ± 15.2, 82% female, 78% Caucasian) we estimated two hierarchical regression models to examine how dimensions of religiousness/spirituality (Brief Multidimensional Measure of Religiousness/Spirituality) and hope (Herth Hope Index) predicted adult life satisfaction (Satisfaction with Life Scale) after controlling for effects of adverse childhood experiences (Adverse Childhood Experiences Study Questionnaire) or chronic sorrow from childhood loss (Kendall Chronic Sorrow Instrument).Results When effects of adverse childhood experiences (ACE) were controlled, higher levels of hope (ß = .416, p < .001) and forgiveness (ß = .273, p = .023) and lower levels of religious/spiritual values/beliefs (ß = -.298, p = .014) predicted higher levels of adult life satisfaction. ACE remained a significant predictor of adult life satisfaction in the final model. When controlling for effects of chronic sorrow, higher life satisfaction was predicted only by forgiveness (ß = .379, p = .003), values/beliefs (ß = -.354, p = .007), and hope (ß = .357, p < .001), with chronic sorrow not a significant predictor of adult life satisfaction in the final model.Conclusion: Greater life satisfaction for adult survivors of childhood loss experiences could be promoted by interventions to enhance hope and foster forgiveness. These adult survivors may also need support as they engage with religious/spiritual struggle to reconcile their values and beliefs with childhood loss.

Vagococcus salmoninarum II-qPCR, tropism and egg-associated transmission.

Vagococcus salmoninarum was identified as the causative agent of a chronic epizootic in broodstock "coaster" brook trout (Salvelinus fontinalis) at the Iron River National Fish Hatchery. The epizootic spanned more than a year, was unresponsive to multiple florfenicol treatments, and resulted in >50% mortality of the affected fish. The decision was made to cull the remaining fish during spawning, which presented an opportunity to more thoroughly examine V. salmoninarum sampling methods, organ tropism and vertical transmission. A newly developed qPCR targeting the pheS gene was used in concert with bacterial culture to show that V. salmoninarum indeed disproportionately affects females and has a tropism for female reproductive tissues. The study demonstrates that some female reproductive tissues (e.g. ovarian fluid, unfertilized eggs) are also an effective option for non-lethal detection. Despite the widespread presence of V. salmoninarum in ovarian fluid and on egg surfaces, we found no evidence of intra-ova transmission.

Vagococcus salmoninarum I-A chronic coldwater streptococcosis in broodstock brook trout (Salvelinus fontinalis) in Wisconsin, USA.

In 2018, Vagococcus salmoninarum was isolated from two lots of broodstock "coaster" brook trout (Salvelinus fontinalis) containing ~1,500 fish at the Iron River National Fish Hatchery, at which time it was identified as the causative agent of a chronic coldwater streptococcosis epizootic. Clinical signs included exophthalmia, lethargy, erratic swimming and loss of equilibrium. Female fish experienced disproportionately higher morbidity and mortality than male co-inhabitants, and routinely retained eggs following spawning. The most consistent gross clinical sign was heart pallor and turbid pericardial effusion. An attempted treatment using florfenicol was ineffective at halting the epizootic, which spanned more than a year and resulted in >50% mortality before remaining fish were culled. As there is no previous documentation of V. salmoninarum at this hatchery or in this species, it is still unclear what circumstances led to this epizootic. The inability to treat this chronic disease led to the loss of valuable broodstock, hampering ongoing fishery conservation efforts in the Great Lakes Basin.

Perceptions of an Obstetric Clinical Rotation by Nursing Students Who Are Men.

BACKGROUND: Maternity nursing is perceived as an unwelcoming specialty to many nursing students who are men. The purpose of this qualitative study was to describe the experiences of nursing students who are men at a Southeastern university before, during, and after their obstetric clinical rotations. METHODS: Students were interviewed individually. The sessions were recorded and transcribed. The transcripts were analyzed individually and collectively by a three-person study team using existential phenomenology. RESULTS: Seven nursing students who are men participated. Four major themes were discovered: preconceptions, welcoming, perceived rejection (of and by participants), and maternity unit culture. Students' perceptions appeared to be grounded in their level of comfort with maternity nursing. CONCLUSIONS: This study adds to the scant amount of information available about experiences of nursing students who are men during their clinical maternity nursing rotation and may help to explain why some men may feel unwelcome in women's health nursing.

Optimizing, validating, and field testing a multiplex qPCR for the detection of amphibian pathogens.

Amphibian populations worldwide are facing numerous threats, including the emergence and spread of infectious diseases. In the past 2 decades, Batrachochytrium dendrobatidis (Bd), a parasitic fungus, and a group of viruses comprising the genus Ranavirus have become widespread and resulted in mass mortality events and extirpations worldwide. In 2013, another novel fungus, B. salamandrivorans (Bsal), was attributed to dramatic declines in populations of fire salamander Salamandra salamandra in the Netherlands. Experimental infections demonstrated that Bsal is highly pathogenic to numerous salamander genera. In an effort to prevent the introduction of Bsal to North America, the US Fish and Wildlife Service (USFWS) listed 201 salamander species as injurious wildlife under the Lacey Act. To determine infection status and accurately assess amphibian health, the development of a sensitive and specific diagnostic assay was needed. We describe the optimization and validation of a multiplex quantitative polymerase chain reaction (qPCR) protocol for the simultaneous detection of Bd, Bsal, and frog virus 3-like ranaviruses. A synthetic genome template (gBlock®) containing the target genes from all 3 pathogens served as the positive control and allowed accurate quantification of pathogen genes. The assay was validated in the field using an established non-lethal swabbing technique to survey local amphibian populations throughout a range of habitats. This multiplex qPCR demonstrates high reproducibility, sensitivity, and was capable of detecting both Bd and ranavirus in numerous locations, species, and life stages. Bsal was not detected at any point during these sampling efforts.

Testing a Model of Sodium Reduction in Hypertensive Older Thai Adults.

Hypertensive older adults will benefit if there is a clear understanding of the factors related to sodium reduction. That would raise awareness of the causes, consequently reducing many health risks, lowering health care costs, and diminishing economic and social burden from high blood pressure. This study explored predictors of urinary sodium excretion. A cross-sectional, correlational study was conducted in 312 hypertensive older Thai adults. Questionnaires related to knowledge, self-care agency, self-care behavior of sodium reduction, and 24-hour urinary sodium analyses were used, followed by the application of structural equation modeling and the Analysis of Moment Structures program. Self-care agency, knowledge, self-care behavior, rural/urban location, and education accounted for 61% of urinary sodium excretion. Self-care agency, knowledge, and self-care behavior were the main predictors in the urinary sodium excretion model. This study suggests establishing supportive educative sodium reduction-related programs that improve knowledge and enhance self-care agency, as well as a comparison of the changes of sodium reduction self-care behavior and urinary sodium excretion over time after the intervention.

Psychotherapy Experiences of Perpetrators of Child Sexual Abuse.

Cynicism about treatment of sex offenders pervades both professional and lay literature. A Cochrane review of randomized controlled trials concluded there is no evidence to support any psychological intervention for sex offenders, but RCT design has limitations for evaluating sex offender treatment. Rarely has a qualitative approach been used to explore perceptions of offenders themselves about their psychotherapy experiences. The purpose of this study was to discover the meaning of therapy experiences to 11 community-dwelling perpetrators of child sexual abuse. They had received therapy during incarceration or after release, or both. Secondary analysis was conducted of phenomenological interviews about participants' early life, during which they spontaneously revealed insights gleaned during therapy in adulthood. Rigor of the analysis was enhanced by reading transcripts aloud and thematizing them in an interdisciplinary interpretive group. Five interrelated themes constituted a gestalt comprising the essence of the therapy experience: "This treatment, it's just totally changed my whole world." Themes included: "It just stripped away all the pretense, all the lies, all the manipulation;" "I didn't understand myself; I found out all about myself through this;" "Nobody knew any of my secrets; that (therapy) was the first time that I got to tell my story;" "The group has become a family for me;" and "I'm very ashamed of what I've done; this treatment has really helped me, gave me a second chance." These findings stand in contrast to cynicism about sex offender therapy and lend support to the increased optimism expressed by several contemporary scholars.

Differential adipose tissue gene expression profiles in abacavir treated patients that may contribute to the understanding of cardiovascular risk: a microarray study.

OBJECTIVE: To compare changes in gene expression by microarray from subcutaneous adipose tissue from HIV treatment naïve patients treated with efavirenz based regimens containing abacavir (ABC), tenofovir (TDF) or zidovidine (AZT). DESIGN: Subcutaneous fat biopsies were obtained before, at 6- and 18-24-months after treatment, and from HIV negative controls. Groups were age, ethnicity, weight, biochemical profile, and pre-treatment CD4 count matched. Microarray data was generated using the Agilent Whole Human Genome Microarray. Identification of differentially expressed genes and genomic response pathways was performed using limma and gene set enrichment analysis. RESULTS: There were significant divergences between ABC and the other two groups 6 months after treatment in genes controlling cell adhesion and environmental information processing, with some convergence at 18-24 months. Compared to controls the ABC group, but not AZT or TDF showed enrichment of genes controlling adherence junction, at 6 months and 18-24 months (adjusted p<0.05) and focal adhesions and tight junction at 6 months (p<0.5). Genes controlling leukocyte transendothelial migration (p<0.05) and ECM-receptor interactions (p = 0.04) were over-expressed in ABC compared to TDF and AZT at 6 months but not at 18-24 months. Enrichment of pathways and individual genes controlling cell adhesion and environmental information processing were specifically dysregulated in the ABC group in comparison with other treatments. There was little difference between AZT and TDF. CONCLUSION: After initiating treatment, there is divergence in the expression of genes controlling cell adhesion and environmental information processing between ABC and both TDF and AZT in subcutaneous adipose tissue. If similar changes are also taking place in other tissues including the coronary vasculature they may contribute to the increased risk of cardiovascular events reported in patients recently started on abacavir-containing regimens.

Psychometric Properties of the Dietary Salt Reduction Self-Care Behavior Scale.

Valid, reliable, and culturally-specific scales to measure salt reduction self-care behavior in older adults are needed. The purpose of this study was to develop the Dietary Salt Reduction Self-Care Behavior Scale (DSR-SCB) for use in hypertensive older adults with Orem's self-care deficit theory as a base. Exploratory factor analysis, Rasch modeling, and reliability were performed on data from 242 older Thai adults. Nine items loaded on one factor (factor loadings = 0.63 to 0.79) and accounted for 52.28% of the variance (Eigenvalue = 4.71). The Kaiser-Meyer-Olkin method of sampling adequacy was 0.89, and the Bartlett's test showed significance (χ2 (df =36) = 916.48, p < 0.0001). Infit and outfit mean squares ranged from 0.81 to 1.25, while infit and outfit standardized mean squares were located at ±2. Cronbach's alpha was 0.88. The 9-item DSR-SCB is a short and reliable scale.

Psychological correlates of HIV-related symptom distress.

This investigation sought to determine the association of symptom distress with selected psychological factors in HIV-infected persons. Data from a randomized controlled trial were used; all subjects who completed baseline data collection were included (N = 99). Data packets included these questionnaires: the Perceived Stress Scale, HIV-related Symptom Distress Scale, and Profile of Mood State. Significant correlations were included in a final regression model. The Perceived Stress Scale, total mood disturbance (including the Profile of Mood State subscales), self-rated current health, and HIV status were independently associated with both frequency of symptoms and symptom distress. Symptom frequency, depression, anger, and fatigue retained significance in the final regression model. Findings from this study indicated significant associations of multiple psychological correlates, suggesting that symptom distress is a complex outcome with a multifactorial etiology. Psychological factors such as depression, anger, and fatigue contribute to the level of distress experienced with HIV-related symptoms.

Childhood sibling and peer relationships of perpetrators of child sexual abuse.

Maltreatment by parents inhibits a child's ability to form relationships with other children. Given that many sex offenders experienced parental abuse, investigation of their relationships with siblings and peers could provide new directions for early childhood interventions. Interview transcripts of 23 community-dwelling perpetrators of sexual abuse against children were examined in this secondary data analysis. Ages of participants ranged from 20s to 70s, and education ranged from GED to PhD. Phenomenological analysis revealed that neither sibling nor peer relationships adequately promoted participants' capacity for developing and maintaining healthy relationships in adulthood. Until societal preventive actions and early clinical interventions are more successful, the crime of child sexual abuse will continue to be far too common.

Childhood experiences of perpetrators of child sexual abuse.

PURPOSE: This study explored the childhood experiences of perpetrators of child sexual abuse. DESIGN AND METHODS: A blend of narrative inquiry and phenomenology was employed. Interviews were conducted with 23 community-dwelling perpetrators (21 males, 2 females). The education of participants ranged from GED to PhD. FINDINGS: There were four types of narratives: There Was No Love; Love Left; Love Was Conflated With Sex; and a Pretty Good Childhood. Chronic sorrow for a painful childhood was evident in most participants. PRACTICE IMPLICATIONS: Cognitive-behavioral treatment may not be optimal for all perpetrators because it is not designed to facilitate recovery from early trauma and loss.

Hearing the silent voices: narratives of health care and homelessness.

Most homeless individuals lack adequate health care. With existing literature as a backdrop, this study sought to understand the experience of homeless persons in the health care system. Using a phenomenological approach, 11 homeless participants were interviewed and the transcripts from these interviews were analyzed for meaning. The health care experiences of the participants could be understood only when viewed within the context of homelessness. The four polar themes that emerged from the analysis--same/different, fair/unfair, freedom/barriers, and choice/no choice--highlighted the great divide between the health care experiences of those with a home and those without. Such understanding can help mental health nurses provide more appropriate care to this population.

Development and validation of the dietary sodium reduction self-care agency scale.

A scale did not exist for measuring the capability or self-care agency of lowering salt consumption in older adults with hypertension. Therefore, our objectives were to develop and validate the Dietary Sodium Reduction Self-Care Agency Scale (DSR-SCA Scale). A 24-item scale was developed and tested in 242 older adults with hypertension. Exploratory factor analysis using principal components extraction, Rasch analysis, and internal consistency reliability were used to evaluate the DSR-SCA Scale. Principal components extraction with Promax rotation was used. An 11-item DSR-SCA Scale with three factor loadings, which measure proficiency, persuasiveness, and resourcefulness, was finalized after it was found to meet the criteria of a minimal factor loading of 0.40, with eigenvalues of 2.20, 1.73, and 1.64, respectively. The Kaiser-Meyer-Olkin was 0.80, and Bartlett's test was significant, χ(2) (df = 55) 403.71, p < 0.0001. The measures accounted for 51% of the total variance. Item infit and outfit mean square ranged from 0.88 to 1.18 and the infit and outfit standardized mean square was -1.8 to 1.7. The 11-item scale demonstrated internal consistency with a Cronbach's alpha coefficient of 0.73 for the total scale. The results showed that the DSR-SCA Scale consisted of three factors that have adequate construct validity and reliability to measure power components and enabling capability related to Orem's self-care theory. This scale is brief, easy-to-complete, and useful for measuring salt reduction self-care agency in older adults with hypertension