Point of Care Analysis of Hematology in the Operating Theater - a Prospective Observational Study of Accuracy and Feasibility.

BACKGROUND: Major surgery entails the risk of severe hemorrhage, and an optimized substitution with red blood cell (RBC) and platelet (PLT) transfusions necessitate rapid test results for RBCs/hemoglobin (HGB)/hematocrit (HCT), and PLTs. The HemoScreen (PixCell Medical, Yokneam Ilit, Israel) is an automated point-of-care hematology analyzer employing image analysis and single-use cuvettes. This study aimed to investigate the correspondence between the HemoScreen and standard laboratory testing (SLT) using the Sysmex XN-9000 in patients undergoing major surgery and to evaluate the feasibility in the operating theater. METHODS: A total of 145 blood samples from 91 adult patients were sampled during abdominal and orthopedic surgery and analyzed on both cell counters. Coefficient of variation (CV) was calculated, Passing-Bablok regression analysis was performed, and Bland-Altman plots were constructed. User experience was assessed through a questionnaire. RESULTS: The HemoScreen showed imprecision with a CV below 5%. Passing-Bablok regression showed positive proportional and negative constant errors for HGB and HCT, a positive proportional error for PLTs, but no dif-ference for RBCs. Bias in the Bland-Altman plots with limits of agreement: RBCs 0.09 x 1012/L (+/- 0.20 x 1012/L), HGB 1.1 g/L (+/- 8.4 g/L), HCT 0.4 % (+/- 2.6%), and PLTs 28.8 x 109/L (+/- 33 x 109/L). The analyzer was scored easy to use with shorter turnaround times compared to SLT. CONCLUSIONS: The HemoScreen is feasible and provides rapid test results with acceptable accuracy for the evaluated application but the two methods cannot be regarded as interchangeable based on the results in this study.

Evaluation of growth, metabolism and production of potentially bioactive components during fermentation of barley with Lactobacillus reuteri.

Eighteen bacterial isolates from millet, buckwheat and rye flour were identified as Lactobacillus reuteri. Genomic fingerprinting (rep-PCR) revealed that they represented five strains and phylogenetic analyses using multi locus sequence analysis (MLSA) showed that all clustered with strains of rodent origin. Two strains (SU12-3 and SU18-3) from different phylogenetic clades were used in fermentations of six varieties of barley, both untreated and heat-treated (with inactivated indigenous enzymes) flour. They were compared with two probiotic strains of human origin (DSM 17938 and ATCC PTA 6475), one previously isolated sourdough strain (LTH 5531) and one strain of Lactobacillus plantarum (36E). Analyses of growth (CFU) and metabolism (1H-NMR) revealed differences at species level, with L. plantarum showing a higher capacity to assimilate nutrients without help of the cereal enzymes. Similarities were observed between L. reuteri strains isolated from sourdough, while the greatest differences between L. reuteri strains were observed between strains 6475 and 17938. Multivariate analysis of the metabolic profiles revealed clear clustering according to flour treatment, species of bacteria and barley variety and to some extent also bacterial strain. Possible bioactive compounds such as γ-aminobutyric acid (GABA), 1,3- propanediol (sign of reuterin production) and histamine were identified and quantified.

Role of Lactobacillus reuteri cell and mucus-binding protein A (CmbA) in adhesion to intestinal epithelial cells and mucus in vitro.

Lactobacillus reuteri, a symbiotic inhabitant of the gastrointestinal tract in humans and animals, is marketed as a probiotic. The ability to adhere to intestinal epithelial cells and mucus is an interesting property with regard to probiotic features such as colonization of the gastrointestinal tract and interaction with the host. Here, we present a study performed to elucidate the role of sortase (SrtA), four putative sortase-dependent proteins (SDPs), and one C-terminal membrane-anchored cell surface protein of Lactobacillus reuteri ATCC PTA 6475 in adhesion to Caco-2 cells and mucus in vitro. This included mutagenesis of the genes encoding these proteins and complementation of mutants. A null mutation in hmpref0536_10255 encoding srtA resulted in significantly reduced adhesion to Caco-2 cells and mucus, indicating involvement of SDPs in adhesion. Evaluation of the bacterial adhesion revealed that of the five putative surface protein mutants tested, only a null mutation in the hmpref0536_10633 gene, encoding a putative SDP with an LPxTG motif, resulted in a significant loss of adhesion to both Caco-2 cells and mucus. Complementation with the functional gene on a plasmid restored adhesion to Caco-2 cells. However, complete restoration of adhesion to mucus was not achieved. Overexpression of hmpref0536_10633 in strain ATCC PTA 6475 resulted in an increased adhesion to Caco-2 cells and mucus compared with the WT strain. We conclude from these results that, among the putative surface proteins tested, the protein encoded by hmpref0536_10633 plays a critical role in binding of Lactobacillus reuteri ATCC PTA 6475 to Caco-2 cells and mucus. Based on this, we propose that this LPxTG motif containing protein should be referred to as cell and mucus binding protein A (CmbA).

Fostering Change Through Occupation-Based Intervention: An International Joint Group Concept Mapping Study.

Further consolidation and clarity regarding occupation as a means to foster change in interventions are needed. The study aimed to utilize the knowledge of occupational scholars to systematically determine what is required to use occupation as means to foster change within occupation-based interventions and to generate a conceptual model from those results. Group Concept Mapping involved the following: preparation, generation of ideas, structuring of statements, data analysis, interpretation of maps, and development of conceptual model. Fifty-two international occupational scholars brainstormed 125 ideas. A cluster rating map with nine clusters posed the foundation for a conceptual model with seven themes, namely, artful use of occupation, evidence-based use of occupation, collaboration to promote occupation, coordinating intervention fit, client factors, sociocultural context, and structural influences. The conceptual model, capturing dimensions and dynamics required for using occupation to foster change, may guide future research into occupation-based interventions.

Extracellular membrane vesicles from Limosilactobacillus reuteri strengthen the intestinal epithelial integrity, modulate cytokine responses and antagonize activation of TRPV1.

Bacterial extracellular membrane vesicles (MV) are potent mediators of microbe-host signals, and they are not only important in host-pathogen interactions but also for the interactions between mutualistic bacteria and their hosts. Studies of MV derived from probiotics could enhance the understanding of these universal signal entities, and here we have studied MV derived from Limosilactobacillus reuteri DSM 17938 and BG-R46. The production of MV increased with cultivation time and after oxygen stress. Mass spectrometry-based proteomics analyses revealed that the MV carried a large number of bacterial cell surface proteins, several predicted to be involved in host-bacteria interactions. A 5'-nucleotidase, which catalyze the conversion of AMP into the signal molecule adenosine, was one of these and analysis of enzymatic activity showed that L. reuteri BG-R46 derived MV exhibited the highest activity. We also detected the TLR2 activator lipoteichoic acid on the MV. In models for host interactions, we first observed that L. reuteri MV were internalized by Caco-2/HT29-MTX epithelial cells, and in a dose-dependent manner decreased the leakage caused by enterotoxigenic Escherichia coli by up to 65%. Furthermore, the MV upregulated IL-1ß and IL-6 from peripheral blood mononuclear cells (PBMC), but also dampened IFN-γ and TNF-α responses in PBMC challenged with Staphylococcus aureus. Finally, we showed that MV from the L. reuteri strains have an antagonistic effect on the pain receptor transient receptor potential vanilloid 1 in a model with primary dorsal root ganglion cells from rats. In summary, we have shown that these mobile nanometer scale MV reproduce several biological effects of L. reuteri cells and that the production parameters and selection of strain have an impact on the activity of the MV. This could potentially provide key information for development of innovative and more efficient probiotic products.

Lactobacilli reduce cell cytotoxicity caused by Streptococcus pyogenes by producing lactic acid that degrades the toxic component lipoteichoic acid.

Lactobacilli are known to prevent colonization by many pathogens; nevertheless, the mechanisms of their protective effect are largely unknown. In this work, we investigated the role of lactobacilli during infection of epithelial cells with group A streptococci (GAS). GAS cause a variety of illnesses ranging from noninvasive disease to more severe invasive infections, such as necrotizing fasciitis and toxic shock-like syndrome. Invasion of deeper tissues is facilitated by GAS-induced apoptosis and cell death. We found that lactobacilli inhibit GAS-induced host cell cytotoxicity and shedding of the complement regulator CD46. Further, survival assays demonstrated that lactic acid secreted by lactobacilli is highly bactericidal toward GAS. In addition, lactic acid treatment of GAS, but not heat killing, prior to infection abolishes the cytotoxic effects against human cells. Since lipoteichoic acid (LTA) of GAS is heat resistant and cytotoxic, we explored the effects of lactic acid on LTA. By applying such an approach, we demonstrate that lactic acid reduces epithelial cell damage caused by GAS by degrading both secreted and cell-bound LTA. Taken together, our experiments reveal a mechanism by which lactobacilli prevent pathogen-induced host cell damage.

Experiences of women with stress-related ill health in a therapeutic gardening program.

BACKGROUND: Stress-related ill health, e.g. burnout, is of great concern worldwide. Effective rehabilitation programs need to be developed and their therapeutic aspects understood. PURPOSE: To explore and describe how women with stress-related ill health who are on sick leave experience the rehabilitation process in a therapeutic garden and how these experiences connect to their everyday lives. METHODS: This longitudinal study used methods from grounded theory. Five women completed three semi-structured interviews at three weekly intervals during rehabilitation and one interview three months after. Data were analyzed using a constant comparative approach. FINDINGS: A secure environment facilitated engagement in activities that provided feelings of enjoyment. These experiences inspired participants to add enjoyable activities in their everyday lives, contributing to occupational balance, despite worries of not be able to continue performing enjoyable activities. Implications. Effective rehabilitation programs need to focus on enjoyable activities in a protective environment to support achievement of occupational balance.

Obesity treatment: a role for occupational therapists?

BACKGROUND: Half of the Danish population is overweight or obese. Obesity can negatively impact health and daily life. The Danish National Board of Health´s guidelines for weight loss programmes to the Danish municipalities, recommends multidisciplinary teams, including occupational therapy, and interventions targeting diet, exercise, psychosocial coping, and everyday life. AIM: To describe the structure and content of obesity programmes offered by the 98 municipalities in Denmark, including details such as the health professionals, programme recipients, dose, structure, content, and the role of occupational therapists. METHOD: A quantitative content analysis was conducted on 234 published Danish municipal weight loss programmes. Programme descriptions were identified through internet searches using both sundhed.dk and Google.com. RESULTS: Various health professionals conducted the programmes, and five involved occupational therapists. Programmes targeted children, adolescent and adults. Dose, structure and content were heterogeneous. CONCLUSION: The majority of the programmes were neither evidence-based, nor did they follow recommendations from Danish National Board of Health. Few programmes addressed the role of habits or social participation. Occupational therapists appear to be under-utilized as providers of the programmes. SIGNIFICANCE: Occupational therapists have a role to play in weight loss programmes, because of their training in activity analysis and their consideration of people, environments, and occupations. These components are included in the recommendations about psychosocial aspects and everyday life from the Danish National Board of Health.

Genome sequence of segmented filamentous bacteria present in the human intestine.

Segmented filamentous bacteria (SFB) are unique immune modulatory bacteria colonizing the small intestine of a variety of animals in a host-specific manner. SFB exhibit filamentous growth and attach to the host's intestinal epithelium, offering a physical route of interaction. SFB affect functions of the host immune system, among them IgA production and T-cell maturation. Until now, no human-specific SFB genome has been reported. Here, we report the metagenomic reconstruction of an SFB genome from a human ileostomy sample. Phylogenomic analysis clusters the genome with SFB genomes from mouse, rat and turkey, but the genome is genetically distinct, displaying 65-71% average amino acid identity to the others. By screening human faecal metagenomic datasets, we identified individuals carrying sequences identical to the new SFB genome. We thus conclude that a unique SFB variant exists in humans and foresee a renewed interest in the elucidation of SFB functionality in this environment.

Lessons learned about occupation-focused and occupation-based interventions: A synthesis using group concept mapping methodology.

Introduction: Occupational therapy (OT) is based on the core assumption that humans are active beings through engagement in occupations. Within OT, occupation is typically used as primary means and/or ends i.e. occupation-focused and/or occupation-based interventions are employed. Studies evaluating such interventions are limited.Objectives: To synthesize experiences about occupation-focused and/or occupation-based interventions. Hence, to identify, organize and prioritize experiences with employing occupation as a core element in OT intervention studies.Methods: Participants were OT PhD students and researchers involved in studies concerning occupation-focused and/or occupation-based interventions. Group Concept Mapping was applied.Results: Based on 133 ideas, a conceptual model emerged encompassing two overall dimensions concerning 'developing interventions' and 'planning intervention studies', respectively. Moreover, ten themes related to one or both dimensions were defined and 94 ideas across clusters had high importance ratings.Conclusion: Synthesis of participants' experiences indicates that 'doing' as agent of change is a core element of OT interventions. Moreover, a multi-level perspective is needed to reach sustainable changes in doing. Group- and peer-support can work as amplifier for change, and flexibility is important during intervention. Such complex interventions need special design and mixed methods in the development, and evaluation of outcome needs to address occupation.

Adherence of clinically isolated lactobacilli to human cervical cells in competition with Neisseria gonorrhoeae.

Lactobacilli are normal inhabitants of our microbiota and are known to protect against pathogens. Neisseria gonorrhoeae is a human specific pathogenic bacterium that colonises the urogenital tract where it causes gonorrhoea. In this study we analysed early interactions between lactobacilli and gonococci and investigated how they compete for adherence to human epithelial cervical cells. We show that lactobacilli adhere at various levels and that the number of adherent bacteria does not correlate to the level of protection against gonococcal infection. Protection against gonococcal adhesion varied between Lactobacillus species. Lactobacillus crispatus, Lactobacillus gasseri and Lactobacillus reuteri were capable of reducing gonococcal adherence while Lactobacillus rhamnosus was not. Lactobacillus strains of vaginal origin had the best capacity to remain attached to the host cell during gonococcal adherence. Further, we show that gonococci and lactobacilli interact with each other with resultant lactobacilli incorporation into the gonococcal microcolony. Hence, gonococci bind to colonised lactobacilli and this complex frequently detaches from the epithelial cell surface, resulting in reduced bacterial colonisation. Also, purified gonococcal pili are capable of removing adherent lactobacilli from the cell surface. Taken together, we reveal novel data regarding gonococcal and lactobacilli competition for adherence that will benefit future gonococcal prevention and treatments.

Global transcriptional response of Lactobacillus reuteri to the sourdough environment.

Lactobacillus reuteri is a lactic acid bacterium that is highly adapted to the sourdough environment. It is a dominant member of industrial type II sourdoughs, and is also able to colonize the intestinal tract of mammals, including humans, and birds. In this study, the transcriptional response of L. reuteri ATCC 55730 was investigated during sourdough fermentation by using whole-genome microarrays. Significant changes of mRNA levels were found for 101 genes involved in diverse cellular processes, such as carbohydrate and energy metabolism, cell envelope biosynthesis, exopolysaccharide production, stress responses, signal transduction and cobalamin biosynthesis. The results showed extensive changes of the organism's gene expression during growth in sourdough as compared with growth in chemically defined medium, and, thus, revealed pathways involved in the adaptation of L. reuteri to the ecological niche of sourdough. The utilization of starch and non-starch carbohydrates, the remodelling of the cell wall, characterized by reduced D-alanylation, and increased amounts of cell wall-associated polysaccharides, as well as the regulatory function of two component systems for cell wall biogenesis and metabolism were suggested by the gene expression data as being important for growth in sourdough. The impact of several L. reuteri genes for effective growth in sourdough was shown by implementation of mutant strains in sourdough fermentation. This study contributes to the understanding of the molecular fundamentals of L. reuteri's ecological competitiveness, and provides a basis for further exploration of genetic traits involved in adaptation to the food environment.

I have mastered the challenge of living with a chronic disease: life stories of people with rheumatoid arthritis.

Our aim with this study was to explore the narrative life story of individuals diagnosed with rheumatoid arthritis. An open qualitative approach, namely narrative biographic methodology, was applied to include the life context of the participants and to deliberately exclude predefinitions of concepts. Ten people with rheumatoid arthritis who retired early because of the disease participated and were interviewed three times according to a narrative biographic interview style. The biographical data and the interview texts were analyzed both individually and in comparison to each other. Some participants regarded rheumatoid arthritis as a challenge for mastery in their lives, whereas others adapted to the disease and "made the best out of a bad situation." Especially in countries where the medical model predominates in health care, our findings can be used to broaden the current view that some health professionals have toward patients, and stress the importance of patients being self-responsible.

Lactobacillus reuteri strains protect epithelial barrier integrity of IPEC-J2 monolayers from the detrimental effect of enterotoxigenic Escherichia coli.

Lactobacillus reuteri is an inhabitant of the gastrointestinal (GI) tract of mammals and birds and several strains of this species are known to be effective probiotics. The mechanisms by which L. reuteri confers its health-promoting effects are far from being fully understood, but protection of the mucosal barrier is thought to be important. Leaky gut is a state of abnormal intestinal permeability with implications for the pathophysiology of various gastrointestinal disorders. Enterotoxigenic Escherichia coli (ETEC) can invade the intestinal mucosa and induce changes in barrier function by producing enterotoxin or by direct invasion of the intestinal epithelium. Our hypothesis was that L. reuteri can protect the mucosal barrier, and the goal of the study was to challenge this hypothesis by monitoring the protective effect of L. reuteri strains on epithelial dysfunction caused by ETEC. Using an infection model based on the porcine intestinal cell line IPEC-J2, it was demonstrated that pretreatment of the cells with human-derived L. reuteri strains (ATCC PTA 6475, DSM 17938 and 1563F) and a rat strain (R2LC) reduced the detrimental effect of ETEC in a dose-dependent manner, as monitored by permeability of FITC-dextran and transepithelial electrical resistance (TEER). Moreover, the results revealed that ETEC upregulated proinflammatory cytokines IL-6 and TNFα and decreased expression of the shorter isoform of ZO-1 (187 kDa) and E-cadherin. In contrast, pretreatment with L. reuteri DSM 17938 and 1563F downregulated expression of IL-6 and TNFα, and led to an increase in production of the longer isoform of ZO-1 (195 kDa) and maintained E-cadherin expression. Interestingly, expression of ZO-1 (187 kDa) was preserved only when the infected cells were pretreated with strain 1563F. These findings demonstrate that L. reuteri strains exert a protective effect against ETEC-induced mucosal integrity disruption.

The impact of time aids on independence and autonomy in adults with developmental disabilities.

The aim of this study was to describe how people with developmental disabilities experienced the use of time aids one year after their introduction by an occupational therapist. Data were obtained through semi-structured interviews. The analysis was performed using a phenomenographic qualitative approach. The results showed that independence and autonomy should be considered as two separate phenomena. Increased independence did not always lead to increased autonomy or vice versa. Four different relationships between these two phenomena were found. They all illustrate the different priorities of the participants and different levels of independence and autonomy. Concerning the usability of time aids, the occupational therapist has to recognize the importance of having frequent communication with the client to understand the phenomena that may affect the use of the aids. The occupational therapist should always try to involve both the client and significant support persons while introducing time aids. Future research in time aids with clients with developmental disabilities could involve multiple interviews with participants combined with participant observations and interviews.

Creativity and order in communication: implications from philosophy to narrative research concerning human occupation.

Narrative theory and methods are increasingly featured in qualitative research relating to human occupation. The present article addresses the concept of narrative in relation to some relevant foundational philosophical and theoretical roots. Specifically a twofold function of narrative as a mechanism for producing order versus creativity is highlighted. It is argued that these functions might serve as resources in research relating to human occupation, specifically regarding issues pertaining to change and agency. Examples from qualitative empirical research relating to human occupation are used to demonstrate points under discussion.

In Vivo and In Vitro Detection of Luminescent and Fluorescent Lactobacillus reuteri and Application of Red Fluorescent mCherry for Assessing Plasmid Persistence.

Lactobacillus reuteri is a symbiont that inhabits the gastrointestinal (GI) tract of mammals, and several strains are used as probiotics. After introduction of probiotic strains in a complex ecosystem like the GI tract, keeping track of them is a challenge. The main objectives of this study were to introduce reporter proteins that would enable in vivo and in vitro detection of L. reuteri and increase knowledge about its interactions with the host. We describe for the first time cloning of codon-optimized reporter genes encoding click beetle red luciferase (CBRluc) and red fluorescent protein mCherry in L. reuteri strains ATCC PTA 6475 and R2LC. The plasmid persistence of mCherry-expressing lactobacilli was evaluated by both flow cytometry (FCM) and conventional plate count (PC), and the plasmid loss rates measured by FCM were lower overall than those determined by PC. Neutralization of pH and longer induction duration significantly improved the mCherry signal. The persistency, dose-dependent signal intensity and localization of the recombinant bacteria in the GI tract of mice were studied with an in vivo imaging system (IVIS), which allowed us to detect fluorescence from 6475-CBRluc-mCherry given at a dose of 1×1010 CFU and luminescence signals at doses ranging from 1×105 to 1×1010 CFU. Both 6475-CBRluc-mCherry and R2LC-CBRluc were localized in the colon 1 and 2 h after ingestion, but the majority of the latter were still found in the stomach, possibly reflecting niche specificity for R2LC. Finally, an in vitro experiment showed that mCherry-producing R2LC adhered efficiently to the intra cellular junctions of cultured IPEC-J2 cells. In conclusion, the two reporter genes CBRluc and mCherry were shown to be suitable markers for biophotonic imaging (BPI) of L. reuteri and may provide useful tools for future studies of in vivo and in vitro interactions between the bacteria and the host.

The cell surface of Lactobacillus reuteri ATCC 55730 highlighted by identification of 126 extracellular proteins from the genome sequence.

Bioinformatical analyses of a draft genome sequence of the commensal bacterium Lactobacillus reuteri ATCC 55730 revealed 126 genes encoding putative extracellular proteins. The function, localization and distribution in bacterial species were predicted. Interestingly, few proteins possessed LPXTG motifs or C-terminal transmembrane anchors. Instead eight proteins were putatively anchored by GW repeats and several secreted proteins were likely to be re-associated to the surface. The majority of the extracellular proteins were widely distributed, i.e., found universally or in gram-positive bacteria, but 24 were only detected in L. reuteri. Further, the number of transporters was lower, while the number of enzyme was higher than in related species.

Antifungal effect of dairy propionibacteria--contribution of organic acids.

Large amounts of food and feed are lost every year due to spoilage by moulds and yeasts. Biopreservation, i.e. the use of microorganisms as preservatives instead of chemicals, has gained increased interest. Lactic acid bacteria and propionibacteria might be particularly useful due to their important role in many food fermentations. Knowledge of the antifungal effects of the organic acids produced by these bacteria is necessary to understand their inhibitory activity. We evaluated the antifungal activity of the type strains of five dairy propionibacteria, Propionibacterium acidipropionici, P. jensenii, P. thoenii, P. freudenreichii subsp. freudenreichii and P. freudenreichii subsp. shermanii against eight food- and feedborne moulds and yeasts. A dual culture system assayed the inhibitory activity on three different agar media, sodium lactate (SL), de Man Rogosa Sharp (MRS) and MRS without acetate (MRS-ac). The amounts of organic acids produced during growth of propionibacteria in liquid SL, MRS and MRS-ac were also determined. The minimal inhibitory concentration (MIC) values of propionic, acetic and lactic acid were established for all fungi at pH 3, 5 and 7. Propionic acid, followed by acetic acid, was the most potent antifungal acid. Inhibition at pH 7 generally required concentrations above 500 mM for all three acids, at pH 5 the MIC values for propionic and acetic acids were 20-120 mM and above 500 mM for lactic acid. At pH 3, the MIC values were, with one exception, below 10 mM for both propionic and acetic acid and above 160 mM for lactic acid. The yeast Pichia anomala was the fungus most resistant to organic acids. The propionibacteria exhibited a pronounced species variation in antifungal activity on MRS (+/-acetate) agar, with P. thoenii being the most potent. Four of the five propionibacteria species produced more propionic and acetic acid in liquid SL medium than in MRS (+/-acetate) broth. However, when SL agar was used as the growth medium, none of the propionibacteria inhibited fungal growth.