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The aim of this literature review was to explore the attitudes of health care workers towards inpatient aggression and to analyse the extent to which attitudes, as defined from a theoretical point of view, were addressed in the selected studies. Databases from 1980 up to the present were searched, and a content analysis was done on the items of the selected studies. The concepts 'cognition' and 'attitude' from the framework of 'The Theory of Reasoned Action' served as categories. The self-report questionnaire was the most common instrument used and three instruments specifically designed to measure attitudes were found. These instruments lacked profound validity testing. From a total of 74 items, two thirds focussed on cognitions and only a quarter really addressed attitudes towards aggression. Research was particularly concerned with the cognitions that nurses had about aggression, and attitudes were studied only to a limited extent. Researchers used different instruments, which makes it difficult to compare results across settings. [ABSTRACT FROM AUTHOR]

At the beginning of the COVID-19 pandemic, diagnostic testing was not accessible for mildly ill or asymptomatic individuals. Military operational circumstances exclude the usage of reference laboratory tests. For that reason, at the beginning of the pandemic alternative test methods were needed in order to gain insight into the SARS-CoV-2 status of military personnel. The objectives of this study are to assess whether SARS-CoV-2 antibody rapid lateral flow assay (LFA) in combination with semi-autonomous SARS-CoV-2 antisense and sense genomic RNA fluorescence in situ hybridization (FISH) could establish disease status in military personnel in a fieldable setting, and to assess how this combination performed and to determine which type of sample performed best. A proof of concept sub-study regarding the SARS-CoV-2 application of the fieldable Biotrack-MED® FISH analyzer, a semi-autonomous multi-sample filter cytometer, preceded this observational prospective cohort pilot study. Dutch military personnel were included in the 26 June 2020-11 May 2021 period. Blood, nasopharyngeal and oropharyngeal swabs and saliva were tested at days 0 and 14. SPSS version 25 descriptive statistics and Cohen's kappa assessed agreement between test methods. Both the sensitivity and specificity of the field tests were calculated with ELISA and PCR as reference. Saliva appeared to be the preferred sample type for FISH, where blood was not useful. FISH analysis and LFA results had a concordance of 42% for testing negative, 30% for recovered from infection, 22% for ongoing-and 58% for acute infection in a reference laboratory lab result outcome (RT-PCR or ELISA respectively). The LFA results on serum and full blood corresponded with the ELISA-obtained results (kappa of 0.61 and 0.63 respectively at day 0 and 0.81 and 0.77 respectively at day 14). LFA (full blood-serum), FISH and RT-PCR on saliva did not reach the 90% sensitivity level advised by WHO, with 64-54, 38 and 71% at day 0 and 80-79, 53 and 24% at day 14 respectively., Competing Interests: GJ is cofounder of the compagny that developed the used automated FISH analyser. MW and JR are working at the Biotrack laboratory. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Wijnberg et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The use of a non-invasive fluorescence in situ hybridization (FISH)-based method on saliva for the detection of SARS-CoV-2 is evaluated in a proof-of-concept study and thereafter utilized in an outpatient setting with the Biotrack-MED® analyzer. For a proof-of-concept study, saliva samples were obtained from 28 persons with mild or moderate COVID-19-related symptoms who were tested RT-PCR positive or negative for SARS-CoV-2. In an outpatient setting, 972 individual saliva samples were utilized. All saliva samples were FISHed with a Cy3-labeled SARS-CoV-2-specific DNA probe and were analyzed manually by fluorescence microscopy (proof-of-concept) or with the SARS-CoV-2 application of the Biotrack-MED® analyzer, a semi-autonomous multi-sample filter cytometer. The proof-of-concept study showed a sensitivity of 96.0% and a specificity of 98.5% and is therefore comparable to the RT-PCR analysis of nasopharyngeal swabs. The outpatient setting showed a sensitivity of 90.9% and a specificity of 94.5% and seems therefore a valid assay for the detection of SARS-CoV-2 in individuals that are healthy, mild or moderate symptomatic. In conclusion, the method evaluated in this study, the FISH-based SARS-CoV-2 application of the Biotrack-MED® analyzer, is a sensitive and reliable assay for the detection of SARS-CoV-2 in the general population., Competing Interests: GGT, GJJ and MW have read the journal’s policy and these authors of this manuscript have the following competing interests: employed by the commercial company Biotrack and patents issued (WO 2010/040371 A1, EP 08874964.3) This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2022 Tamminga et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: In this study, we examined the predictive values of a moral deliberate and paternalistic attitude on the propensity of yielding to pressure. In these hypothesised positive and negative relationships, we further sought to ascertain whether moral disengagement plays a pivotal role when individuals deviate from ethical standards, rules and regulations when yielding to pressure., Aim(s): This study's primary aim was to assess the predictive value of a moral deliberative and paternalistic attitude for yielding to pressure when physician assistants (PAs) and nurse practitioners (NPs) face moral conflicts., Method: This validation study was cross-sectional and based on a convenience sample of Dutch PAs and NPs. The MSQ-DELIB and MSQ-PATER scales indicate a moral deliberate or paternalistic attitude. These scales were assumed to have a predictive value towards the degree of yielding to pressure by PAs and NPs. Yielding to pressure was measured by two vignettes in which respondents faced a moral conflict (vignette 1: prescribing unindicated antibiotics and vignette 2: discharging a difficult patient from the hospital)., Results: Only moral deliberation was a significant predictor of yielding to pressure. That is, we found a positive effect in vignette 1 (in which the pressure came from the patient). In contrast, we found a negative relationship in vignette 2 (in which pressure went from the working environment). Paternalism did not affect yielding to pressure in either vignette., Conclusion: This study suggests that PAs and NPs having a moral deliberative attitude makes them receptive to pressure exerted by patients to break moral standards. On the other hand, they are more resilient against doing so when this pressure comes from different sources than the patient. Further research is needed to find more conclusive evidence for this differential effect., (© 2021 The Authors. Scandinavian Journal of Caring Sciences published by John Wiley & Sons Ltd on behalf of Nordic College of Caring Science.)

Saliva is a matrix which may act as a vector for pathogen transmission and may serve as a possible proxy for SARS-CoV-2 contagiousness. Therefore, the possibility of detection of intracellular SARS-CoV-2 in saliva by means of fluorescence in situ hybridization is tested, utilizing probes targeting the antisense or sense genomic RNA of SARS-CoV-2. This method was applied in a pilot study with saliva samples collected from healthy persons and those presenting with mild or moderate COVID-19 symptoms. In all participants, saliva appeared a suitable matrix for the detection of SARS-CoV-2. Among the healthy, mild COVID-19-symptomatic and moderate COVID-19-symptomatic persons, 0%, 90% and 100% tested positive for SARS-CoV-2, respectively. Moreover, the procedure allows for simultaneous measurement of viral load ('presence', sense genomic SARS-CoV-2 RNA) and viral replication ('activity', antisense genomic SARS-CoV-2 RNA) and may yield qualitative results. In addition, the visualization of DNA in the cells in saliva provides an additional cytological context to the validity and interpretability of the test results. The method described in this pilot study may be a valuable diagnostic tool for detection of SARS-CoV-2, distinguishing between 'presence' (viral load) and 'activity' (viral replication) of the virus. Moreover, the method potentially gives more information about possible contagiousness., Competing Interests: WJBvW and IDW have declared that no competing interests exist. GJJ and MW have read the journal’s policy and these authors of this manuscript have the following competing interests: employment by the commercial company Biotrack and patents issued (WO 2010/040371 A1, EP 08874964.3). This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Rationale, Aims, and Objectives: The current study and previous research have called the six-component model of Lützen's 30-item Moral Sensitivity Questionnaire (MSQ) into question. For this reason, we re-examined the construct validity of this instrument., Methods: In this cross-sectional study, which was based on a convenience sample of Dutch nurse practitioners (NPs) and physician assistants (PAs), we tested the validity of MSQ items using exploratory and confirmatory factor analyses (EFA and CFA, respectively)., Results: The EFA revealed a two-component model, which was then tested as a target model with CFA and was found to have good model fit. Some items were correlated with two uncorrelated latent constructs, which we labelled as "paternalistic" and "deliberate" attitudes towards patients., Conclusions: As in previous studies, the analyses in the current study, which was conducted among PAs and NPs, did not reveal six dimensions for the 30 items. Two new latent dimensions of moral sensitivity were psychometrically tested and confirmed. These two components relate to studies investigating ethical behaviour, and they can be used to describe the moral climate in healthcare organizations. The scales are indicators of the extent to which health professionals behave in a deliberate (sensitive) or paternalistic (insensitive) manner towards the opinions of patients within the context of medical decision-making., (© 2020 The Authors. Journal of Evaluation in Clinical Practice published by John Wiley & Sons Ltd.)

In this study, an alternative parameter for quantifying the signals of fluorescently labelled bacteria (e.g. propidium iodide, Cyanine 3, etc.) in microscopic images was investigated. Three common parameters (mean grey value (MGV), mean grey value which is corrected for the background (MGVcwB) and the signal to background ratio (SBR) per bacterial cell) are used as reference parameters. As an alternative, the coefficient of variation (CV) is defined as the ratio of the logarithm of the standard deviation and the logarithm of the mean grey value of a bacterial cell in a microscopic image. The actual fluorescence value was safeguarded by measuring commercially available fluorescence latex microspheres at regular time intervals within our study. The precision and the correlation of the respective values of MGV, MGVcwB, SBR and CV taken from identical images were measured and subsequently normalized in order to enhance the inter-parameter comparability. The average precision of CV was the highest (89% ± 14) with decreasing numbers for MGVcwB, SBR, and MGV (78% ± 25, 71% ± 32, and, 52% ± 22, respectively). Changes in operational parameters, e.g., microscope settings, protocol steps, etc., yielded good results for the CV but less precise results for MGV, MGVcwB, and SBR in the analyses of identical images. In conclusion, using the alternative parameter CV, changes in the composition of microbial ecosystems may thus be investigated at the highest precision level., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Aim: To explore the direct and indirect effect of the personality meta-traits 'Stability' and 'Plasticity' on moral reasoning among nurse practitioners (NPs) and physician assistants (PAs)., Background: Moral reasoning is influenced by being prone to moral disengagement and personality traits. Moral disengagement is observed among professionals in many fields, including healthcare providers. Moral disengagement is known to be provoked by environmental stressors and influenced by certain personality traits., Design: A cross-sectional approach was used including self-report questionnaires., Methods: A convenience sample of Dutch NPs (N = 67) and PAs (N = 88) was surveyed via online questionnaires between January and March 2015, using (a) the Defining Issues Test; (b) the BIG five inventory; and (c) the Moral Disengagement Scale. Structural equation modelling (SEM) was employed for estimating the construct validity of two meta-traits of personality and to test unidirectional influences on moral reasoning., Results: Only the Stability trait was a direct predictor of moral reasoning whereas both Stability and Plasticity were precursors of moral disengagement. Both personality meta-traits had statistically significant indirect effects on moral reasoning through a low level of moral disengagement. The influence of both personality traits on the level of moral reasoning was increased by strong self-censure on entering into morally disengaged interactions., Conclusion: The personality meta-trait 'Stability' is an indicator of moral reasoning and is explained by a lower propensity to morally disengage among highly stable people. Although the meta-trait Plasticity exerts an indirect effect through moral disengagement on moral reasoning, it is not a direct indicator of moral reasoning., (© 2018 John Wiley & Sons Ltd.)

Background: Previous studies have shown that nursing students' perceptions of nursing change over time. Little research has been undertaken in the Netherlands of students entering nursing programmes and of how they progress., Objectives: The aims of this study were to explore whether nursing students' orientation and attitudes towards nursing changed over time, when these changes occurred, and what factors influenced the changes. We also aimed to identify the factors which prompted them to consider leaving their programmes, and what factors affected their motivation to stay., Design: The study used a longitudinal quantitative design., Participants: Questionnaires were administered to all students enrolled in a Bachelor's of Nursing programme at four nursing universities of applied sciences in the Netherlands (n=1414). The data for this study were collected during the first two years of the programme, from September 2011 to June 2013. A total of 123 respondents completed the survey each year and this group was used to examine changes over time., Methods: At four time intervals respondents completed a survey consisting of 1) the Nursing Orientation Tool, 2) the Nursing Attitude Questionnaire and 3) background characteristics. Non-parametric tests were used to explore changes in factor scores over time., Results: The results showed an improvement in the students' orientation and attitudes towards knowledge, skills and the professional roles of nurses, while empathic behaviour decreased over time. Although the changes showed non-linear patterns over time, the results showed clear effects between the different time points. The reasons for attrition (24%) proved to be related both to problems with the educational programme and to personal problems. An important motivator for students to stay in the course was their passionate desire to become nurses, suggesting that the positive aspects of a nursing career dominated the problems they encountered., Conclusions: Tutors and mentors should pay more attention to the individual perceptions and problems of first and second-year students, both in the classroom and during clinical placements. Knowledge of the students' perceptions from the very beginning could be vital to study success., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

State of the art software methods (such as fixed value approaches or statistical approaches) to create a binary image of fluorescent bacterial cells are not as accurate and precise as they should be for counting bacteria and measuring their area. To overcome these bottlenecks, we introduce biological significance to obtain a binary image from a greyscale microscopic image. Using our biological significance approach we are able to automatically count about the same number of cells as an individual researcher would do by manual/visual counting. Using the fixed value or statistical approach to obtain a binary image leads to about 20% less cells in automatic counting. In our procedure we included the area measurements of the bacterial cells to determine the right parameters for background subtraction and threshold values. In an iterative process the threshold and background subtraction values were incremented until the number of particles smaller than a typical bacterial cell is less than the number of bacterial cells with a certain area. This research also shows that every image has a specific threshold with respect to the optical system, magnification and staining procedure as well as the exposure time. The biological significance approach shows that automatic counting can be performed with the same accuracy, precision and reproducibility as manual counting. The same approach can be used to count bacterial cells using different optical systems (Leica, Olympus and Navitar), magnification factors (200× and 400×), staining procedures (DNA (Propidium Iodide) and RNA (FISH)) and substrates (polycarbonate filter or glass)., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Aim: To explore the focus of nurse practitioners on health care in terms of cure and care., Background: Nurse practitioners are expected to act on the intersection of cure and care. However, in clinical practice and education, a clear model covering this area is lacking; therefore, it is unknown to what extent nurse practitioners are focused on this specific area. Graduate theses may reflect the focus of nurse practitioners., Design: Sequential exploratory mixed method., Methods: In total, 413 published abstracts of graduate theses of a Dutch Master of Advanced Nursing Practice (2000-2015) were analysed using the International Classification of Functioning, Disability and Health. Data source included aim, question and outcome of each thesis and graduates' characteristics. A qualitative deductive approach was used for the analyses. Theses were classified as focused on cure, care, or on the intersection of cure and care., Results: A small majority of 53% (N = 219) of the graduate theses addressed patient's health status and could be classified in the International Classification of Functioning, Disability and Health. Of the classified theses, 48% were focused on cure, 39% on the intersection of cure and care and 13% on care. While the percentage of theses addressing health status increased significantly over the 15-year period, the percentage of theses focused on cure, care and on the intersection of cure and care remained the same., Conclusion: The graduate theses reflected that nurse practitioners are increasingly oriented towards patients' health status. However, their focus is predominantly on cure rather than on the intersection of cure and care., (© 2016 John Wiley & Sons Ltd.)

Aims and Objectives: To research the use of ambiguous language in written information concerning patients' functioning and to identify problems resulting from the use of ambiguous language in clinical practice., Background: Many projects that aimed to introduce standard terminology concerning patients' functioning in clinical practice are unsuccessful because standard terminology is rarely used in clinical practice. These projects mainly aim to improve communication by reducing ambiguous language. Considering their lack of success, the validity of the argument that language ambiguity is used in clinical practice is questioned., Design: An integrative literature review., Methods: A systematic search of the MEDLINE (1950-2012) and CINAHL (1982-2012) databases was undertaken, including empirical and theoretical literature. The selected studies were critically appraised using a data assessment and extraction form., Results: Seventeen of 767 papers were included in the review and synthesis. The use of ambiguous language in written information concerning patients' functioning was demonstrated. Problems resulting from the use of ambiguous language in clinical practice were not identified. However, several potential problems were suggested, including hindered clinical decision-making and limited research opportunities., Conclusion: The results of this review demonstrated the use of ambiguous language concerning patients' functioning, but health professionals in clinical practice did not experience this issue as a problem. This finding might explain why many projects aimed at introducing standard terminology concerning functioning in clinical practice to solve problems caused by ambiguous language are often unsuccessful. Language ambiguity alone is not a valid argument to justify the introduction of standard terminology., Relevance to Clinical Practice: The introduction of standard terminology concerning patients' functioning will only be successful when clinical practice requires the aggregation and reuse of data from electronic patient records for different purposes, including multidisciplinary decision-making and research., (© 2014 John Wiley & Sons Ltd.)

Aims and Objectives: To compare a functioning assessment based on the International Classification of Functioning, Disability and Health (ICF) with a conventional medical assessment, in terms of their respective consequences for health professionals' clinical decision-making and the fit with patient's own perspective of health., Background: In chronic diseases, pathogenic-oriented health care falls short in generating all the information required for determining healthcare provision to improve health. A broader, so-called salutogenic approach, by using the ICF, focusing on how to stay healthy, rather than on what causes diseases, seems more appropriate., Design: A cross-sectional comparative study using data from a randomised controlled trial., Methods: Data about patient problems and professional healthcare activities were collected from a total of 81 patients with severe multiple sclerosis who were randomly assigned to one of two groups: the ICF group, assessed with a functioning assessment (n = 43), and the medical group, assessed with a conventional medical assessment (n = 38). Data were analysed statistically using descriptive and inferential statistics., Results: A functioning assessment resulted in the registration of significantly more patient problems in the health components 'participation' and 'environmental factors', as well as significantly more professional healthcare activities befitting these components. The ICF group had a significant positive correlation between registered problems by health professionals and patients' self-reported problems, whereas the medical group had several negative correlations., Conclusion: A functioning assessment resulted in a care plan that not only was broader and more complete but also reflected the patients' self-reported problems more closely than a medical assessment, without a loss of focus on medical problems., Relevance to Clinical Practice: This study has shown that some health problems remain unnoticed by a medical assessment alone, which is especially important for the chronically ill. A functioning assessment provides a strong foundation for identifying all relevant information related to health., (© 2013 John Wiley & Sons Ltd.)

Data were collected from 121 staff members (20 direct support staff teams) on background characteristics of the individual staff members and their teams (gender, age, years of work experience, position and education), the frequency and form of aggression of clients with an intellectual disability (verbal or physical), staff members' attitudes towards aggression, and the types of behavioural interventions they executed (providing personal space and behavioural boundary-setting, restricting freedom and the use of coercive measures). Additionally, client group characteristics (age of clients, type of care and client's level of intellectual disability) were assessed. Multilevel analyses (individual and contextual level) were performed to examine the relations between all studied variables and the behavioural interventions. The results showed that for providing personal space and behavioural boundary-setting as well as for restricting freedom, the proportion of variance explained by the context (staff team and client group characteristics) was three times larger than the proportion of variance explained by individual staff member characteristics. For using coercive measures, the context even accounted for 66% of the variance, whereas only 8% was explained by individual staff member characteristics. A negative attitude towards aggression of the direct support team as a whole proved to be an especially strong predictor of using coercive measures. To diminish the use of coercive measures, interventions should therefore be directed towards influencing the attitude of direct support teams instead of individual staff members., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Little is currently known about the attitudes of psychiatric nurses toward patient aggression, particularly from an international perspective. Attitudes toward patient aggression of psychiatric nurses from five European countries were investigated using a recently developed and tested attitude scale. Data were collected from a convenience sample of 1,769 student nurses and psychiatric nurses. Regression analysis was performed to identify personal and occupational characteristics of the respondents able to predict their attitude toward aggression. Analysis of variance was used to identify significant differences in attitudes between and among countries. Attitude was predicted by sex, contractual status (full vs. part time), and the type of ward on which subjects worked. With one exception (communicative attitude), attitudes differed across countries. More research on attitude formation is needed to determine which factors account for these differences.

One of the factors known to be associated with the management of patient aggression is the attitude of staff members towards the aggressive behaviour of patients. The construct validity of an instrument measuring the attitudes of staff towards inpatient aggression in psychiatry was evaluated in this international multi-centre study. Factor analysis and simultaneous component analysis were performed with data from a convenience sample of 1769 psychiatric nurses working in psychiatric hospitals and student nurses from nursing schools. The samples were recruited by fellow researchers in their home country. The original 32-item version (POAS) was reduced to 18 items comprising five attitude scales with solid psychometric properties. The types of attitudes were labelled offensive, communicative, destructive, protective and intrusive. The format of the correlations between the types of attitudes suggested the existence of two basic underlying divergent domains in the scale. The 'communication' and 'protection' scale components on the one hand, and the 'offence', 'destruction' and 'intrusion' components on the other. The five types of attitude proved to be invariant across samples from five European countries. The Aggression Scale (ATAS) is a reliable and valid measure that will enable researchers to perform international comparative research on attitudes and aggression.

The applicability of whole-cell hybridization for the identification of pathogenic bacteria in blood from septic patients was examined. Oligonucleotide probes, fluorescently labeled with fluorescein isothiocyanate, directed against the variable regions of the 16S rRNAs of the following bacterial species and/or genera were used: Streptococcus spp., Enterococcus faecalis, Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Escherichia coli, Pseudomonas aeruginosa, and the Enterobacteriaceae family. A probe specific for the rRNAs of almost all bacteria and its complementary, reversed counterpart was used as positive and negative control, respectively. The probes were used in conjunction with a fast and simple-to-use protocol for whole-cell hybridization. This protocol yields an identification after 25 to 45 min, depending on whether the bacterium is gram positive or gram negative. A total of 182 blood samples which tested positive in a blood culture machine were investigated. All probes except for the ones for S. aureus and the CoNS showed sensitivities and specificities of 1.000. It was concluded that whole-cell hybridization is well suited for the fast screening of septic blood containing streptococci and/or enterococci or gram-negative rods.

An automated microscopy-based method using fluorescently labelled 16S rRNA-targeted oligonucleotide probes directed against the predominant groups of intestinal bacteria was developed and validated. The method makes use of the Leica 600HR image analysis system, a Kodak MegaPlus camera model 1.4 and a servo-controlled Leica DM/RXA ultra-violet microscope. Software for automated image acquisition and analysis was developed and tested. The performance of the method was validated using a set of four fluorescent oligonucleotide probes: a universal probe for the detection of all bacterial species, one probe specific for Bifidobacterium spp., a digenus-probe specific for Bacteroides spp. and Prevotella spp. and a trigenus-probe specific for Ruminococcus spp., Clostridium spp. and Eubacterium spp. A nucleic acid stain, 4',6-diamidino-2-phenylindole (DAPI), was also included in the validation. In order to quantify the assay-error, one faecal sample was measured 20 times using each separate probe. Thereafter faecal samples of 20 different volunteers were measured following the same procedure in order to quantify the error due to individual-related differences in gut flora composition. It was concluded that the combination of automated microscopy and fluorescent whole-cell hybridisation enables distinction in gut flora-composition between volunteers at a significant level. With this method it is possible to process 48 faecal samples overnight, with coefficients of variation ranging from 0.07 to 0.30.

Six 16S rRNA-targeted oligonucleotide probes were designed, validated, and used to quantify predominant groups of anaerobic bacteria in human fecal samples. A set of two probes was specific for species of the Bacteroides fragilis group and the species Bacteroides distasonis. Two others were designed to detect species of the Clostridium histolyticum and the Clostridium lituseburense groups. Another probe was designed for the genera Streptococcus and Lactococcus, and the final probe was designed for the species of the Clostridium coccoides-Eubacterium rectale group. The temperature of dissociation of each of the probes was determined. The specificities of the probes for a collection of target and reference organisms were tested by dot blot hybridization and fluorescent in situ hybridization (FISH). The new probes were used in initial FISH experiments to enumerate human fecal bacteria. The combination of the two Bacteroides-specific probes detected a mean of 5.4 x 10(10) cells per g (dry weight) of feces; the Clostridium coccoides-Eubacterium rectale group-specific probe detected a mean of 7.2 x 10(10) cells per g (dry weight) of feces. The Clostridium histolyticum, Clostridium lituseburense, and Streptococcus-Lactococcus group-specific probes detected only numbers of cells ranging from 1 x 10(7) to 7 x 10(8) per g (dry weight) of feces. Three of the newly designed probes and three additional probes were used in further FISH experiments to study the fecal flora composition of nine volunteers over a period of 8 months. The combination of probes was able to detect at least two-thirds of the fecal flora. The normal biological variations within the fecal populations of the volunteers were determined and indicated that these variations should be considered when evaluating the effects of agents modulating the flora.

The influence of oral treatment with a suspension of non-pathogenic Escherichia coli cells (commercially available as: Symbioflor II) on the morphological composition of the gut microflora and on the systemic humoral immune response (the IgG-, IgA- and IgM-isotype) against the bacterial cells in the Symbioflor II preparation was measured. After a pretreatment period of 21 days, ten healthy human volunteers ingested 1*10(8) cells of E. coli daily for 14 days. Thereafter a follow-up period of 28 days completed the study. The results of this study indicated that no effect of the treatment on the composition of the gut microflora could be observed. However, the immune-fluorescence measurements revealed a significant increase in circulating amounts of IgG directed against the administered E. coli cells. It is concluded that the treatment only resulted in a specific humoral immune response, while the gut microflora is not modulated.

Background: The composition of a sample of faecal bacteria can be determined by culturing different dilutions on specific media. However, not all bacteria can be cultured and media are not always specific. With a culture-independent approach a more accurate picture of the composition of the intestinal flora may be obtained., Methods: Fluorescently labelled oligonucleotide probes targeted at 16S ribosomal RNA sequences specific for a bacterial genus were designed and applied for fluorescence in situ hybridization (FISH) of bacteria in human faecal samples., Results: The mean number of Bifidobacterium spp. and the total number of anaerobic bacteria per gram of faeces were determined by culturing and with the probe technique. Although in both cases the number of Bifidobacterium spp. was about the same, 2.38 x 10(9) and 2.45 x 10(9), it was found that the contribution of Bifidobacterium spp. to the total composition is overestimated due to the lower number of total anaerobic bacteria estimated by culturing., Conclusion: Genus-specific or group-specific fluorescent 16S rRNA probes may become an invaluable tool in gut ecology studies.

Three 16S rRNA hybridization probes were developed and tested for genus-specific detection of Bifidobacterium species in the human fecal flora. Variable regions V2, V4, and V8 of the 16S rRNA contained sequences unique to this genus and proved applicable as target sites for oligodeoxynucleotide probes. Determination of the genus specificity of the oligonucleotides was performed by whole-cell hybridization with fluorescein isothiocyanate-labelled probes. To this end, cells were fixed on glass slides, hybridized with the probes, and monitored by videomicroscopy. In combination with image analysis, this allowed quantification of the fluorescence per cell and objective evaluation of hybridization experiments. One of the probes developed was used to determine the population of Bifidobacterium spp. in human fecal samples. A comparison was made with results obtained by cultural methods for enumeration. Since both methods gave similar population estimates, it was concluded that all bifidobacteria in feces were culturable. However, since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration.

The association of plasma histidine-proline-rich glycoprotein (HPRG) with plasminogen (PLG) was examined using a sucrose density gradient assay in order to evaluate the effects of several relevant conditions on complex formation. Addition of PLG shifts the S-value of 125I-labeled HPRG from 4.8S to 6.8S, providing the first direct evidence that HPRG associates with the zymogen form of plasmin in solution. Complex formation is not sensitive to pH in the range of pH 6.5-8.5, but is abolished at high ionic strength (1 M NaCl). No species differences were found, as either rabbit or human HPRG bound readily to rabbit or human PLG. Of the ligands of HPRG tested, mesoheme (20 microM) but not heparin (M(r) 10,000, 10 microM) inhibits the formation of the HPRG-PLG complex. Modification of lysine residues of HPRG or competitive binding by lysine and anti-fibrinolytic agents containing primary amino groups also inhibits association. Conversely, modification of arginine or histidine residues of HPRG has no effect on complex formation. These results indicate that HPRG has independent binding sites for heparin and PLG and confirm that one or more lysine residues of HPRG are involved in its recognition by PLG. The protein-protein interaction was also quantitatively characterized at thermodynamic equilibrium by analytical ultracentrifugation. The stoichiometry and dissociation constant (KD) of the complex were determined from the equilibrium distribution of fluorescein-isothiocyanate-labeled PLG in the presence of HPRG. The experimental data were analyzed by nonlinear least-squares curve fitting and indicated that a heterodimer is formed.(ABSTRACT TRUNCATED AT 250 WORDS)

Several techniques that use computer analysis of microscopic images have been developed to study the complicated microbial flora in the human intestine, including measuring the shape and fluorescence intensity of bacteria. These techniques allow rapid assessment of changes in the intestinal flora and could apply equally to other complex microbial ecosystems.

The purpose of the study was to identify NANDA-nursing diagnoses with psychiatric patients who are deaf or hard of hearing. The research study took place at the VIA (Visual Interaction Ward) of the Robert Fleury Stichting at Leidschendam. By means of focused interviews with nurses data were gathered from which, using content analysis, nursing diagnoses were identified. These diagnoses, consisting of defining characteristics and related factors, were reviewed on their validity by the nurses who take care of these patients. In this way an inventory was made up of the diagnoses most characteristic of the population. From the data, the diagnosis of impaired communication related to deafness was generated by the researchers themselves. Although this diagnosis is not mentioned in literature, it was supported by the respondents who participated in the study.

The use of polyclonal antibodies in differentiating between samples of faecal microflora derived from ten healthy volunteers was assessed. The distribution of FITC-labelled antibodies (of the IgA, IgG and IgM isotype) over 144 morphologically distinct subsets of faecal bacteria ('morphotypes') was quantified by means of digital morphometry and quantitative immunofluorescence. Furthermore, a new dataprocessing algorithm was developed which makes objective quantitation of the antibody distributions over the faecal morphotypes possible. The results of this study imply that the antibody binding capacity of faecal morphotypes is a unique characteristic of faecal microflora.

Membranes isolated from porcine aortic endothelial cells (PAEC) contain a CoA-independent transacylase enzyme (CoA-IT). CoA-IT, an integral membrane protein, transfers an acyl moiety to added [3H]alkylhydroxyglycerophosphocholine (LPAF) to generate [3H]alkylacylglycerophosphocholine (alkylacyl-GPC). This enzyme exhibits an apparent Km of 0.7 microM and a Vmax of 0.8 nmol/min per mg for the transfer of an acyl group to added [3H]LPAF. The addition of the nonionic detergent Triton X-100 (TX-100) (0.5 mg/ml), the sulfhydryl reagents N-ethylmaleimide (NEM) (200 microM) or thimerosal (200 microM), or pre-incubating the membranes at 95 degrees C for 10 min all decreased LPAF: CoA-IT activity by more than 95%. The inhibitory action of NEM or thimerosal suggests that sulfhydryl group(s) are involved in or are close to the catalytic site of LPAF: CoA-IT.