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Immunochemical methods for environmental monitoring.

Immunochemical methods for environmental analysis must be taken into consideration more for their ability to expand the potential of analytical measures rather than for substituting current methodologies. Moreover, the full potential of these methods has yet to be realized. Indeed, the terms and concepts of immunology are new to most analytical chemists, even if environmental science has always been an interdisciplinary field. On the other hand, the clinical development of immunoassays means that much experience has been gained in the analysis of blood, urine, and tissue samples. The immunochemical analysis of samples from soils, ground water, waste chemicals, poses new challenges in sample preparation that have yet to be extensively studied, and in the future there may be immunoassays better suited for the particular problems associated with environmental monitoring.

Environmental Monitoring↗

Parental decision making in pediatric cancer end-of-life care: using focus group methodology as a prephase to seek participant design input.

The ultimate aim of our research program is to provide strategies that facilitate parental decision-making for parents of children with cancer receiving end-of-life care. As a first step to develop this program, we needed insight into parents' reactions and opinions about the research methods planned for a larger study. In particular, we needed their opinions about the general experience of making the decision between palliative cytotoxic chemotherapy and supportive care alone and the factors that parents regard as important when making this decision. In addition, we wished to know whether the methodology proposed for the future study was easy to understand and whether it might cause unnecessary emotional trauma. Finally, we asked their opinions regarding the appropriate target sample of parents to include in the future study. Qualitative data about these issues were collected using focus group methodology involving seven participants. The comments made during the focus group discussions were content-analyzed for common themes. The results from the focus group discussion led to particular modifications in the proposed design and interview strategies planned for the future larger study. We found it was extremely beneficial to include a focus group pre-phase in a study that will interview parents in a high sensitivity area.

Adaptation, Psychological↗

Sensory evaluation of drinking water by consumer panels.

The human senses play an important role in assessing the quality of food and of the environment. Particularly the chemical senses of taste and smell determine the pleasantness of foods and drinks and may provide a warning mechanism for the presence of more or less toxic contaminants. Even after the recent rapid development of powerful analytical techniques the human nose can easily detect trace amounts of chemicals at levels many times lower than the analytical detection limits. Although taste and odour assessment of drinking water has been practiced in many waterworks laboratories by small panels, relatively unreliable results were obtained and such sensory data played up till now only a minor role in the management of the water treatment plant. More sophisticated and reliable methods for sensory water quality evaluation are discussed. Special attention is given to the characteristics of the human senses of taste and smell, to which the methodology of sensory assessment should be better adapted. High numbers of observations on a sample are generally needed, for which purpose the use of large panels has to be realized. In this respect the help of large consumer panels is an alternative for the presently often used small laboratory groups. As an example the preliminary results of a large ongoing experiment of sensory water quality assessment by 2 consumer panels of 100 persons each in the Rotterdam area are discussed. Drinking water quality is judged at the consumer homes every week. The experiment started September 1979 and will be continued for one year.

Humans↗

Interobserver reliability of the click test: a rapid bedside test to determine surfactant function.

OBJECTIVE: To determine the interobserver reliability of the click test, a rapid bedside test of surfactant function. METHODOLOGY: One hundred samples (48 amniotic fluid, 28 gastric aspirates, 24 tracheal aspirates) were obtained from pregnant women, term and preterm infants managed at two perinatal centres. Gestational ages of the pregnancies or infants at time of sampling ranged from 24 to 40 weeks (mean +/- s.d.: 31+/-5). The click test was performed independently by two observers, with differing levels of experience, and blinded to each other's results. Interobserver reliability was determined for samples classified as negative (no clicking bubbles), equivocal (a few clicking bubbles seen after 2 min of looking), and positive (clicking bubbles seen easily on first look). As negative and equivocal test results suggest surfactant dysfunction, these results were also combined to provide a dichotomised result. RESULTS: For samples classified as negative, equivocal and positive the interobserver agreement was 0.83, with agreement beyond chance shown by a quadratic weighted Kappa of 0.74 (95% CI = 0.62-0.86). When negative and equivocal results were combined, the simple Kappa was 0.73 (95% CI = 0.59-0.87). CONCLUSIONS: The click test has a high level of interobserver reliability. It is rapid, simple, accurate and able to be taught to medical and nursing staff with appropriate training. It has many potential applications including the prediction of fetal lung maturity in women with premature rupture of membranes, and in term and preterm infants who require the bedside testing of surfactant function.

Amniotic Fluid↗

The problem drinker's lived experience of suffering: an exploration using hermeneutic phenomenology.

Research in the area of problem drinking has traditionally relied on quantitative methodologies which view the problem from the researcher's perspective. The purpose of this hermeneutic-phenomenological study was to describe and understand the problem drinker's lived experience of suffering using a philosophy and research approach which preserves the uniqueness of the experience from the sufferer's point of view. The method involved conducting in-depth interviews with a sample of six problem drinkers. Interviews were analysed using an interpretive process, which aimed at generating a deeper understanding of the topic by facilitating a fusion of the world views of both participant and researcher. A reflexive journal recorded the involvement of the self of the researcher throughout the research process. Suffering was viewed as a spiralling vicious circle of physical, psychological, social and spiritual distress. Symptoms of physical dependence, shame and guilt emerged strongly as being both sequelae of heavy drinking and cues to further drinking bouts. Evoking memories of previous suffering through telling one's story was found to be an empowering and motivating force. The results have relevance to specialist and generic workers, who are urged to pay greater attention to the social, psychological and spiritual care of problem drinkers.

Adult↗

Epidemiological and clinical aspects of bipolar disorders: controversies or a common need to redefine the aims and methodological aspects of surveys.

Data from surveys of large samples showed the lifetime prevalence rates of bipolar disorder around 1.5%. A main question is whether the low prevalence rates of bipolar disorders are not an artefact of the over-diagnosis of depression and under-diagnosis of bipolar-II.Analysis of the clinician's logical inferential diagnostic process, confirms that the patient does not represent the sole source of useful information because many patients do not experience hypomania as distress but rather as recovery from depression or as a period during which they felt truly well.Epidemiological data are derived from interviews carried out by lay staff which only reflect the patient's point of view.The clinical monitoring study carried out alongside the ESEMED project found for the diagnosis of mood disorders, a Kappa agreement (versus clinical interview) which ranged from 0.23 in Spain to 0.49 in France.If we consider exactly what a Kappa of 0.4 implies for a disorder with an "identified" prevalence rate of 2%, we discover that the prevalence rate may have been under-diagnosed approximately 1.5-fold, so 67% of cases may not have been identified and 50% of the identified cases may be false positives.It is legitimate to surmise that the prevalence reported by recent (extremely costly) epidemiological surveys may be doubtful.Which direction should epidemiology take in dealing with the serious matter of bipolar disorders?Recently, some community surveys were carried out in the USA using the Mood Disorder Questionnaire. In the ensuing debate, one side claimed that the instrument was scarcely accurate when used in the general population, gave rise to numerous false positives and that the high prevalence reported was therefore a mere artefact. The other side defended the results reported by the research studies, on the basis that "positive" cases were homogeneous with regard to the high level of subjective distress, low social functioning and employment and with the high recourse to health care structures.It is quite probable that the problem lies at the root of the matter, in the definition of the gold standard.In the present state of our knowledge on course and response to treatment, the current diagnostic thresholds applied for mixed states and hypomanic episodes seem to be unsatisfactory.It is inconceivable that the diagnostic gold standard should be determined only on the basis of a structured interview of patients alone. But unless there is clinical consensus on the diagnostic threshold for hypomania and mixed states, there can be no consensus on the findings of epidemiological research.

Editorial↗

Methodology of trials with antiemetics.

Chemotherapy-induced nausea and vomiting can today be controlled with the "new" antiemetics or with their various combinations in a high percentage of patients. Despite this, for some subgroups of patients, certain chemotherapy regimens and some aspects of the phenomenon (delayed presentation), emesis remains a critical problem. Hence, there is the necessity for a continuous effort in the search for new drugs or better treatment modalities and the absolute requisite that these efforts be carried out according to a sound and verified trial methodology. Nausea and vomiting induced by antineoplastic agents are extremely variable phenomena, depending not only on the characteristics of chemotherapy regimens and of the patient population, but also on a subjective feeling generated by the impact of the care system on the patient's individual situation. Therefore, precisely because of this high variability, large comparative trials should be carried out to ensure that the sample is sufficiently representative for the most efficacious antiemetic regimen to be detected. In this field, some of the main problems arising in all clinical trials have their own specificity, particularly the study design, whether completely randomized or cross-over, the follow-up and the importance of prognostic factors. Among these, age, gender and previous chemotherapy with experience of nausea and vomiting have been confirmed as important. In addition, some topics must clearly be highlighted: the definition of the response variables and the assessment of variability of the results obtained with the same antiemetic regimen from one cycle of chemotherapy to the next (i.e. persistence). The greater attention devoted today to delayed emesis raises some methodological questions: this paper suggests some possible solutions for a better evaluation of this phenomenon.

Antiemetics↗

NMR spectroscopy in inhomogeneous B0 and B1 fields with non-linear correlation.

Resolved NMR spectra from samples in inhomogeneous B0 and B1 fields can be obtained with the so-called "ex situ" methodology, employing a train of composite or adiabatic z-rotation RF pulses to periodically refocus the inhomogeneous broadening during the detection of the time-domain signal. Earlier schemes relied on a linear correlation between the inhomogeneous B0 and B1 fields. Here the pulse length, bandwidth, and amplitude of the adiabatic pulses of the hyperbolic secant type are adjusted to improve the refocusing for a setup with non-linear correlation. The field correlation is measured using a two-dimensional nutation experiment augmented with a third dimension with varying RF carrier frequency accounting for off-resonance effects. The pulse optimization is performed with a computer algorithm using the experimentally determined field correlation and a standard adiabatic z-rotation pulse as a starting point for the iterative optimization procedure. The shape of the z-rotation RF pulse is manipulated to provide refocusing for the conditions given by the sample-, magnet-, and RF-coil geometry.

2-Propanol↗

SELDI-TOF-MS of saliva: methodology and pre-treatment effects.

Interest in saliva as a diagnostic fluid for monitoring general health and for early diagnosis of disease has increased in the last few years. In particular, efforts have focused on the generation of protein maps of saliva using advanced proteomics technology. Surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) is a novel high throughput and extremely sensitive proteomic approach that allows protein expression profiling of large sets of complex biological specimens. In this study, large scale profiling of salivary proteins and peptides, ranging from 2 to 100kDa was demonstrated using SELDI-TOF-MS. Various methodological aspects and pre-analytical variables were analysed with respect to their effects on saliva SELDI-TOF-MS profiling. Results show that chip surface type and sample type (unstimulated versus stimulated) critically affect the amount and composition of detected salivary proteins. Factors that influenced normal saliva protein profiling were matrix composition, sample dilution and binding buffer properties. Delayed processing time experiments show certain new peptides evolving 3h post-saliva donation, and quantitative analyses indicate relative intensity of other proteins and peptides changing with time. The addition of protease inhibitors partly counteracted the destabilization of certain protein/peptide mass spectra over time suggesting that some proteins in saliva are subject to digestion by intrinsic salivary proteases. SELDI-TOF-MS profiles also changed by varying storage time and storage temperature whereas centrifugation speed and freeze-thaw cycles had minimal impact. In conclusion, SELDI-TOF-MS offers a high throughput platform for saliva protein and peptide profiling, however, (pre-)analytical conditions must be taken into account for valid interpretation of the acquired data.

Biomarkers↗

The effects of atomic force microscopy upon nominated living cells.

This work describes a system for precise re-location of cells within a monolayer after atomic force imaging. As we know little about probe interaction with soft biological surfaces any corroborative evidence is of great importance. For example, it is of paramount importance in living cell force microscopy that interrogated cells can be re-located and imaged by other corroborative technologies. Methodologies expressed here have shown that non-invasive force parameters can be established for specific cell types. Additionally, we show that the same sample can be transferred reliably to an SEM. Results here indicate that further work with live cells should initially establish appropriate prevailing force parameters and that cell damage should be checked for before and after an imaging experiment.

Animals↗

Adult health status of women with histories of childhood abuse and neglect.

PURPOSE: Several recent studies have found associations between childhood maltreatment and adverse adult health outcomes. However, methodologic problems with accurate case determination, appropriate sample selection, and predominant focus on sexual abuse have limited the generalizability of these findings. SUBJECTS AND METHODS: We administered a survey to 1,225 women who were randomly selected from the membership of a large, staff model health maintenance organization in Seattle, Washington. We compared women with and without histories of childhood maltreatment experiences with respect to differences in physical health status, functional disability, numbers and types of self-reported health risk behaviors, common physical symptoms, and physician-coded ICD-9 diagnoses. RESULTS: A history of childhood maltreatment was significantly associated with several adverse physical health outcomes. Maltreatment status was associated with perceived poorer overall health (ES = 0.31), greater physical (ES = 0.23) and emotional (ES = 0.37) functional disability, increased numbers of distressing physical symptoms (ES = 0.52), and a greater number of health risk behaviors (ES = 0.34). Women with multiple types of maltreatment showed the greatest health decrements for both self-reported symptoms (r = 0.31) and physician coded diagnoses (r = 0.12). CONCLUSIONS: Women with childhood maltreatment have a wide range of adverse physical health outcomes.

Adult↗

Multiple-quantum relaxation dispersion NMR spectroscopy probing millisecond time-scale dynamics in proteins: theory and application.

New relaxation dispersion experiments are presented that probe millisecond time-scale dynamical processes in proteins. The experiments measure the relaxation of (1)H-(15)N multiple-quantum coherence as a function of the rate of application of either (1)H or (15)N refocusing pulses during a constant time relaxation interval. In contrast to the dispersion profiles generated from more conventional (15)N((1)H) single-quantum relaxation experiments that depend on changes in (15)N((1)H) chemical shifts between exchanging states, (1)H-(15)N multiple-quantum dispersions are sensitive to changes in the chemical environments of both (1)H and (15)N spins. The resulting multiple-quantum relaxation dispersion profiles can, therefore, be quite different from those generated by single-quantum experiments, so that an analysis of both single- and multiple-quantum profiles together provides a powerful approach for obtaining robust measures of exchange parameters. This is particularly the case in applications to protonated proteins where other methods for studying exchange involving amide proton spins are negatively influenced by contributions from neighboring protons. The methodology is demonstrated on protonated and perdeuterated samples of a G48M mutant of the Fyn SH3 domain that exchanges between folded and unfolded states in solution.

Magnetic Resonance Spectroscopy↗

Room for improvement? Reporting response rates and recruitment in nursing research in the past decade.

AIMS: This paper reports an analysis of recruitment and response rates in published nursing research in three peer reviewed nursing journals in 2002. We wished to establish if the deficits in reporting nursing research identified a decade earlier had been addressed. BACKGROUND: This analysis was informed by our personal experiences of research which produced widely differing response rates. An examination of the literature revealed a lack of consensus on desirable response rates in nursing research. Previous analyses have shown deficits in describing participants, sampling methods and reporting recruitment. METHODS: Papers reporting empirical research in three nursing journals in 2002 were reviewed in terms of a number of variables including research methodology, respondent type, recruitment method, response rate, location, and data collection method. Nominal coding was used as necessary. Data were analysed using the Statistical Package for Social Sciences and a variety of descriptive statistics were employed. RESULTS: Half of the papers did not report a response rate. Of those which did, over three-quarters of both qualitative and quantitative studies had response rates of 60% or more. Research conducted in hospital and educational settings produced higher response rates than those in community settings. Studies with response rates of less than 60% did not always refer to their rates in the study limitations, and low response rates do not appear to be a barrier to publication. CONCLUSION: Reporting of sampling, recruitment and response rates in nursing research must be improved to support nursings' claim to be an evidence-based profession and to underpin clinical governance requirements. Only through improvements in the quality of nursing research publications can knowledge be extended and a better-informed research community be created.

Bibliometrics↗

A guide to the use of focus groups in health care research: Part 1.

Focus group interviewing is becoming a popular research method among nurses as well as allied health professionals. Prior to performing a focus group interview a novice must first search the literature to gain an understanding of the function, uses, preparation, conduct and analysis of this form of methodology. This paper has been compiled in an attempt to synthesise, condense and simplify the information from an extremely wide and varied focus group literature and provide a comprehensive review to assist nurses who are considering utilising this methodology in their research studies. The purpose of this paper was stimulated by one of the author's (MP) own experience learning about focus group methodology before commencing her first focus group.

Bias↗

Ameliorating the tension: use of forgiveness for healing.

PURPOSE/OBJECTIVES: To generate a grounded substantive theory of the process of forgiveness in patients with cancer. DESIGN: Qualitative approach, grounded theory. SETTINGS: Inpatient hospice, cancer support groups, homecare patients. SAMPLE: 25 adult patients with cancer between the ages of 35 and 88 (X = 62.4 years): 13 men and 12 women receiving active, palliative, or terminal treatment. METHODS: Using grounded theory methodology, open-ended interviews were transcribed verbatim and analyzed using constant comparative analysis and expert and participant validation. MAIN RESEARCH VARIABLE: Forgiveness as reflected in participants' past and current experiences. FINDINGS: Participants described hurtful events that initially caused severe negative emotional responses. Participants also described attempts to focus on living out priority values prompted by their cancer diagnosis. An escalating tension ensued between maintaining negative emotions and acting on personal values. Forgiveness was used to relieve this tension. Using several methods to gain perspective allowed resolving these negative emotions and living out priority values to various degrees. CONCLUSIONS: A cancer diagnosis promotes the rethinking of life priorities regardless of the stage of disease. For these participants, forgiveness was used to help clarify personal values and to help eliminate negative emotions from an inflicted hurt. IMPLICATIONS FOR NURSING PRACTICE: Nurses can facilitate the healing process by helping patients to identify and clarify priority values, recognizing appropriate definitions of forgiveness, using active listening or referral to help patients analyze their situations, encouraging active approaches to forgiving, and encouraging perseverance in the process.

Adaptation, Psychological↗

Cytochemical detection systems for in situ hybridization, and the combination with immunocytochemistry, 'who is still afraid of red, green and blue?'.

An overview is given of the different non-radioactive cytochemical detection methodologies that are currently utilized to localize nucleic acid sequences in chromosomes, cells and tissue sections. Dependent on the reporter molecule (fluorochrome, enzyme or hapten) that is used to modify the appropriate nucleic acid probe, and the sensitivity that is required, the in situ hybridized sequences can be detected either directly after hybridization or indirectly, using cytochemical detection and amplification layers. These may then contain antibody and/or avidin molecules conjugated to fluorochromes, enzymes or colloidial gold particles. Since the choice of a suitable probe-labelling method in combination with a fluorescence, enzyme cytochemical or immunogold-silver detection procedure is often determined by the user's own practical experience and applications, the different detection methodologies are compared with each other in detail with respect to sensitivity, resolution, applicability for multiple probe detection, and signal evaluation. Furthermore, procedures are reviewed for the combination of in situ hybridization with immunocytochemical detection of proteins and/or incorporated bromodeoxyuridine, which allow the simultaneous visualization of genomic phenotypic and/or cell cycle parameters in the same sample. Possible improvements with respect to sensitivity, specificity and multiplicity of the detection methods, which may be interesting for one's own experimental design, are finally being discussed.

Animals↗

An exploration of tutors' experiences of facilitating problem-based learning. Part 1--an educational research methodology combining innovation and philosophical tradition.

The use of problem-based learning (PBL) in Health Professional curricula is becoming more wide spread. Although the way in which the tutor facilitates PBL can have a major impact on students' learning (Andrews and Jones 1996), the literature provides little consistency as to how the tutor can effectively facilitate PBL (Haith-Cooper 2000). It is therefore important to examine the facilitation role to promote effective learning through the use of PBL. This article is the first of two parts exploring a study that was undertaken to investigate tutors' experiences of facilitating PBL. This part focuses on the methodology and the combining of innovative processes with traditional philosophical traditions to develop a systematic educational research methodology. The study was undertaken respecting the philosophy of hermeneutic phenomenology but utilised alternative data collection and analysis technique. Video conferencing and e-mail were used in conjunction with more traditional processes to access a worldwide sample. This paper explores some of the issues that arose when undertaking such a study. The second article then focuses on exploring the findings of the study and their implications for the facilitation of PBL.

Attitude of Health Personnel↗