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PubMed2005

Managing health inequalities locally: a baseline survey of primary care trusts' experience with health equity audit in the implementation year.

Health equity audit (HEA), a pragmatic policy tool to ensure that services and resources are focused on issues that have the highest impact on health inequalities, has now become embedded in the national strategy to tackle these inequalities through a new mandatory requirement that primary care trusts (PCTs) conduct one such audit annually. This study aimed to assess their experience through a national baseline survey in 2004, all PCTs (n = 303) being electronically mailed a questionnaire with non-respondent follow-up. Replies were received from 132 PCTs (44%), a representative sample of PCT diversity, most of whom had completed only a few steps in the audit cycle. Audit topics (most frequently coronary heart disease, smoking cessation, and access issues) and dimensions of inequity (mainly area and deprivation) were agreed through only limited engagement with local strategic partnerships. Local public health networks and multiagency teams were infrequent partners in undertaking the HEA. Most PCTs wanted comparator data, good practice examples, and specific methodological expertise. While significant progress has been made, this survey shows only limited use of HEA as a tool for multisectoral use by PCTs in partnership with others and a focus on intra-PCT comparisons at the expense of those with a wider pool of 'look-a-likes'.

Data Collection↗
PubMed2000

Johne's disease and milk: do consumers need to worry?

Mycobacterium paratuberculosis, an acid-fast bacillus that causes enteritis in ruminants, has been suggested as an etiological agent of Crohn's disease in humans. The mode of transmission is unclear; however, some evidence suggests that humans may become infected via contaminated milk. Currently, it is not known whether commercial pasteurization effectively kills M. paratuberculosis in contaminated raw milk. Using a laboratory-scale pasteurizer unit designed to simulate the high-temperature, short-time method (72 degrees C, 15 sec) currently used by commercial dairies, we previously demonstrated that treatment of raw milk inoculated with 10(4) to 10(6) cfu of M. paratuberculosis/ml reduced numbers to an undetectable level. However, M. paratuberculosis is an intracellular pathogen that resides within the macrophages of the host and evades destruction. We subsequently performed further experiments examining heat treatment of milk inoculated with mammary gland macrophages containing ingested M. paratuberculosis. Heat treatment of these samples under high-temperature, short-time conditions demonstrated that the macrophage does not protect the organism because we were unable to recover any viable M. paratuberculosis from the samples. Conversely, other researchers have demonstrated that a residual population of M. paratuberculosis may survive heat treatment of milk. In addition, a recent news report stated that viable M. paratuberculosis organisms have been cultured from retail-ready milk in Ireland. A summary of past and current studies concerning this issue along with a discussion of methodologies used to recover M. paratuberculosis from experimentally inoculated milk will be presented in this paper.

Animals↗
PubMed1998

The development of a scale to measure concepts of schizophrenia: experience among Brazilian psychiatrists.

INTRODUCTION: Among psychiatric disorders schizophrenia is often said to be the condition with the most disputed definition. The Bleulerian and Schneiderian approaches have given rise to diagnostic formulations that have varied with time and place. Controversies over the concept of schizophrenia were examined within European/North American settings in the early 1970s but little has since been reported on the views of psychiatrists in developing countries. In Brazil both concepts are referred to in the literature. A scale was developed to measure adherence to Bleulerian and Schneiderian concepts among psychiatrists working in S. Paulo. METHODOLOGY: A self-reported questionnaire comprising seventeen visual analogue-scale statements related to Bleulerian and Schneiderian definitions of Schizophrenia, plus sociodemographic and training characteristics, was distributed to a non-randomised sample of 150 psychiatrists. The two sub-scales were assessed by psychometric methods for internal consistency, sub-scale structure and test-retest reliability. Items selected according to internal consistency were examined by a two-factor model exploratory factor analysis. Intraclass correlation coefficients described the stability of the scale. RESULTS: Replies were received from 117 psychiatrists (mean age 36 (SD 7.9)), 74% of whom were made and 26% female. The Schneiderian scale showed better overall internal consistency than the Bleulerian scale. Intra-class correlation coefficients for test-retest comparisons were between 0.5 and 0.7 for Schneiderian items and 0.2 and 0.7 for Bleulerian items. There was no negative association between Bleulerian and Schneiderian scale scores, suggesting that respondents may hold both concepts. Place of training was significantly associated with the respondent's opinion; disagreement with a Bleulerian standpoint predominated for those trained at the University of S. Paulo. CONCLUSIONS: The less satisfactory reliability for the Bleulerian sub-scale limits confidence in the whole scale but on the other hand this questionnaire contributes to the understanding of the controversy over Bleulerian and Schneiderian models for conceptualisation of schizophrenia, the former requiring more inference and therefore being prone to unreliability.

Adult↗
PubMedDate not supplied

African American women coping with breast cancer: a qualitative analysis.

PURPOSE/OBJECTIVES: To determine how African American women cope with breast cancer. DESIGN: Descriptive and exploratory study. SAMPLE/SETTING: 66 African American women diagnosed with breast cancer were interviewed in the southeastern United States. METHODS: Data were collected through tape-recorded interviews using a semistructured interview guide. Data were analyzed by content analysis and frequency distributions. MAIN RESEARCH VARIABLES: Coping strategies used by women to adapt to a diagnosis of breast cancer. FINDINGS: Coping strategies described by African American women included relying on prayer, avoiding negative people, developing a positive attitude, having a will to live, and receiving support from family, friends, and support groups. CONCLUSION: Spirituality played a major role in these African American women coping with breast cancer. Supportive networks also served as a vital asset throughout the breast cancer experience. Participants discussed the need for culturally sensitive breast cancer support groups. IMPLICATIONS FOR NURSING: Nurses must recognize coping strategies that African American women with breast cancer use. Healthcare professionals need to develop culturally sensitive breast cancer support groups. Throughout the breast cancer experience, nurses must assess communication patterns among African American families. Nurses should serve as healthcare advocates for African American women with breast cancer.

Adaptation, Psychological↗
PubMed2002

Inverse 15N-metabolic labeling/mass spectrometry for comparative proteomics and rapid identification of protein markers/targets.

The inverse labeling/mass spectrometry strategy has been applied to protein metabolic (15)N labeling for gel-free proteomics to achieve the rapid identification of protein markers/targets. Inverse labeling involves culturing both the perturbed (by disease or by a drug treatment) and control samples each in two separate pools of normal and (15)N-enriched culture media such that four pools are produced as opposed to two in a conventional labeling approach. The inverse labeling is then achieved by combining the normal (14)N-control with the (15)N-perturbed sample, and the (15)N-control with the (14)N-perturbed sample. Both mixtures are then proteolyzed and analyzed by mass spectrometry (coupled with on-line or off-line separation). Inverse labeling overcomes difficulties associated with protein metabolic labeling with regard to isotopic peak correlation and data interpretation in the single-experiment approach (due to the non-predictable/variable mass difference). When two data sets from inverse labeling are compared, proteins of differential expression are readily recognized by a characteristic inverse labeling pattern or apparent qualitative mass shifts between the two inverse labeling analyses. MS/MS fragmentation data provide further confirmation and are subsequently used to search protein databases for protein identification. The methodology has been applied successfully to two model systems in this study. Utilizing the inverse labeling strategy, one can use any mass spectrometer of standard unit resolution, and acquire only the minimum, essential data to achieve the rapid and unambiguous identification of differentially expressed protein markers/targets. The strategy permits quick focus on the signals of differentially expressed proteins. It eliminates the detection ambiguities caused by the dynamic range of detection. Finally, inverse labeling enables the detection of covalent changes of proteins responding to a perturbation that one might fail to distinguish with a conventional labeling experiment.

Amino Acid Sequence↗
PubMed2006

Methodological considerations for the use of stable isotope probing in microbial ecology.

Stable isotope probing (SIP) is a method used for labeling uncultivated microorganisms in environmental samples or directly in field studies using substrate enriched with stable isotope (e.g., (13)C). After consumption of the substrate, the cells of microorganisms that consumed the substrate become enriched in the isotope. Labeled biomarkers, such as phospholipid-derived fatty acid (PLFA), ribosomal RNA, and DNA can be analyzed with a range of molecular and analytical techniques, and used to identify and characterize the organisms that incorporated the substrate. The advantages and disadvantages of PLFA-SIP, RNA-SIP, and DNA-SIP are presented. Using examples from our laboratory and from the literature, we discuss important methodological considerations for a successful SIP experiment.

Biodiversity↗
PubMed1994

Verbal autopsies for adult deaths: issues in their development and validation.

BACKGROUND: The verbal autopsy appears to be an attractive technique for the assessment of causes of adult death in settings where the proportion of people who die while under medical care is low. Verbal autopsies have been used extensively to assess causes of childhood deaths. The existing experience in verbal autopsies for adult deaths is limited mainly to maternal deaths. This paper aims to highlight the critical issues in verbal autopsies to assess causes of adult death which need further research. METHODS: This paper reviews the methods of verbal autopsy used in 35 published studies and discusses issues in the development of verbal autopsies, including mortality classification, design of questionnaires, interviewers, respondents, recall periods, procedures for deriving a diagnosis and the recording of single versus multiple causes of death. It also discusses issues in the validation of verbal autopsies, including the choice of reference diagnosis and the required sample size. RESULTS: The methodological approaches used in verbal autopsy studies have varied widely. Very few studies of the validity of verbal autopsies have been conducted. In these studies, the reported sensitivity and specificity varied widely between different causes of death and between studies. CONCLUSIONS: The information available is inadequate to draw firm conclusions on preferred methodological approaches for verbal autopsies for adult deaths. Before these tools are used more widely for adult deaths, further research is required to compare alternative methods and to evaluate the validity of this tool in a range of settings.

Adolescent↗
PubMed1997

Exploring the psychosocial meaning of recurrent cancer: a descriptive study.

Many persons diagnosed with malignancy will experience one or more recurrences of malignancy. Little is known about the psychosocial meaning of recurrent cancer. Using Lazarus and Folkman's model of stress, appraisal and coping, the two purposes of this descriptive study were to (a) describe the meaning of a recurrence of cancer to the patient and (b) to explore if the patient perceives the diagnosis of recurrence as being different from the initial diagnosis of cancer. Purposeful sampling for persons with recent recurrent malignancy produced a sample (n = 20). Subjects completed an unstructured, indepth interview. The meaning of the recurrence was influenced by prior cancer-related experiences of the subjects and dominated by death and death-related concerns. Differences from the initial diagnosis included a deeper awareness of the significance of the "cancer diagnosis".

Adaptation, Psychological↗
PubMed2000

Sampling extracellular aspartate, glutamate and gamma-aminobutyric acid in striate cortex of awake cat by in vivo microdialysis: surgical and methodological aspects.

A method which permits repeated microdialysis in the cortical layers of area 17 of the awake cat is described. Under visual control through a surgical microscope and using a stereotactic instrument, four probe guides are permanently implanted in area 17 of one hemisphere of the anesthetized animal and two fixation bars are mounted on the skull to allow fixation of the cat in a stereotactic frame. The implantation of four probe guides in the same hemisphere allows simultaneous sampling from different cortical regions serving different parts of the visual field. A removable transparent cover protects the probe guides. After recovery from surgery the awake cats are trained to adapt to a fixation of 5 h in a stereotaxic apparatus. Once adapted to that situation, the cats are ready for microdialysis experiments without anesthesia. The day of the experiment, the awake animal was fixed in the stereotactic frame and the probes inserted into the guides. To test the validity of the method, the basal efflux and the depolarization efflux, triggered by the addition of 65 mM K(+) to the artificial cerebrospinal fluid, of the amino acids aspartate, glutamate and gamma-aminobutyric acid are measured by two HPLC-electrochemical detection methods. The exact localization of the probes and the reaction of the surrounding tissue is studied using immunocytochemistry for glutamate and glial fibrilary acidic protein. Our neurochemical and morphological results suggest the feasibility of multiple and repeated probe insertions for microdialysis experiments in the cerebral cortex of awake and behaving cat. This method provides a new tool to investigate the cortical plasticity.

Animals↗
PubMed1988

Failure to alter sympathoadrenal response to psychosocial stress following aerobic training.

This experiment assessed the impact of aerobic training on sympathoadrenal, heart rate and subjective responses to psychosocial stress. Subjects were six previously sedentary males who demonstrated marked improvement in fitness level following 10 weeks of training. Plasma samples, heart rate and subjective arousal ratings were obtained prior to, during, and following exposure to stressful mental tasks within a laboratory session. With the exception of training-related bradycardia which was manifest throughout the session, no changes in plasma epinephrine and norepinephrine concentrations, heart rate or subjective reactivity or recovery were seen. These findings are inconsistent with the hypothesized beneficial effects of aerobic training on stress-response. Methodological factors such as self-selection inherent in previous correlation work or the duration of the training program should be considered in this context. The potential contribution of training-related bradycardia to enhanced coping with challenging situations is explored.

Adult↗
PubMed2006

Analysis of human hippocampus gangliosides by fully-automated chip-based nanoelectrospray tandem mass spectrometry.

Modern microfluidic devices are currently introduced in electrospray (ESI) mass spectrometry (MS), tending to substitute the classical capillary-based ESI infusion. Automated systems using the combination of robotized sample handling and chip-based ESI are significantly increasing the analysis reproducibility, precision, throughput, and efficiency. In the last couple of years our group developed the chip-based ESI-MS approach for glycomics in biomedical research and applied it for oligosaccharide, glycopeptide and ganglioside investigation. Here we report upon the optimization and application of this modern technique for the analysis of differential ganglioside expression patterns in human fetal and adult hippocampus. By this methodology, ganglioside species exhibiting high degree of heterogeneity in the ceramide motifs and biologically-relevant modifications could be identified in human hippocampus. The ultra-high reproducibility of the experiments uniquely provided by the chip-ESI approach allowed for a reliable MS-based ganglioside comparative assay. Moreover, the particular feature of chip ESI-tandem MS to provide structural information at high sensitivity was useful for detailed characterization of hippocampus-associated species. The experimental data presented in this study indicate the benefits of microfluidic/MS for determination of the topospecific brain ganglioside composition and development-related changes in their expression, which might be of high value in clinical investigation and for studies related to ganglioside-based therapy of central nervous system diseases.

Adult↗
PubMed2006

Characterisation of an industrial affinity process used in the manufacturing of digoxin-specific polyclonal Fab fragments.

This paper describes the effect of several variables on the affinity process for the production of the FDA approved biotherapeutic product Digoxin Immune Fab (Ovine) (DigiFab, Protherics Inc., TN, USA). The study considers the effects of column re-use on matrix capacity and on the subsequent recovery of the antibody product, and the impact of varying column loading on matrix performance. The methodology used could be equally applied to assess the feasibility of using an affinity matrix for commercial scale purification of alternative antibody derived biotherapeutics. The capacity and specific Fab recovery were calculated through 24h equilibrium and mass balance studies. Results were assessed against data obtained through confocal scanning laser microscopy. Scale-down experiments produced specific Fab recoveries and purities that were comparable with those at production scale. The matrix capacity was found to be 45+/-15 mg of Fab/ml of matrix. Through the use of fluorescent DigiFab and confocal scanning techniques, Fab uptake onto single affinity bead was evaluated. Average intensity values calculated for each sample provided direct real-time, measure of Fab binding and matrix capacity. The results suggest that the affinity matrix had a limited reuse life as a drop in recovery is observed following the completion of a small number of process cycles (30% after three runs). The findings support that which is seen at the current manufacturing scale, where the affinity column is used for a limited number of runs. Results from this study can be used as a basis for future optimisation of this purification process.

Animals↗
PubMed2003

Measurement uncertainty from physical sample preparation: estimation including systematic error.

A methodology is proposed, which employs duplicated primary sampling and subsequent duplicated physical preparation coupled with duplicated chemical analyses. Sample preparation duplicates should be prepared under conditions that represent normal variability in routine laboratory practice. The proposed methodology requires duplicated chemical analysis on a minimum of two of the sample preparation duplicates. Data produced from the hierarchical design is treated with robust analysis of variance (ANOVA) to generate uncertainty estimates, as standard uncertainties ('u' expressed as standard deviation), for primary sampling (ssamp), physical sample preparation (sprep) and chemical analysis (sanal). The ANOVA results allow the contribution of the sample preparation process to the overall uncertainty to be assessed. This methodology has been applied for the first time to a case study of pesticide residues in retail strawberry samples. Duplicated sample preparation was performed under ambient conditions on two consecutive days. Multi-residue analysis (quantification by GC-MS) was undertaken for a range of incurred pesticide residues including those suspected of being susceptible to loss during sample preparation procedures. Sampling and analytical uncertainties dominated at low analyte concentrations. The sample preparation process contributed up to 20% to the total variability and had a relative uncertainty (Uprep%) of up to 66% (for bupirimate at 95% confidence). Estimates of systematic errors during physical sample preparation were also made using spike recovery experiments. Four options for the estimation of measurement uncertainty are discussed, which both include and exclude systematic error arising from sample preparation and chemical analysis. A holistic approach to the combination and subsequent expression of uncertainty is advised.

Calibration↗
PubMedDate not supplied

Social separation, housing relocation, and survival in simian AIDS: a retrospective analysis.

OBJECTIVE: To test the hypothesis that changes in housing, particularly those involving social separations, would have a negative impact on survival in rhesus monkeys experimentally inoculated with the simian immunodeficiency virus (SIV). METHODS: An archival methodology was used. Colony records at four Regional Primate Research Centers were screened, and data pertaining to demographics, contents of the inoculum, medical history before and after inoculation, and housing relocations and social companions were coded. The final sample size totaled 298 individuals. RESULTS: Following statistical control of covariates, housing relocations and social separations in the 90-day period before SIV inoculation and in the 30-day period after inoculation were associated with decreased survival. There was evidence that housing disruptions occurring earlier after inoculation were associated with shorter survival. Finally, a subset of animals was found to have been socially housed after SIV inoculation; such experience had negative consequences for survival. CONCLUSIONS: The results indicate that psychosocial experiences that likely produce a stressful state are associated with shorter survival in SIV-infected monkeys.

Animals↗
PubMed1993

Marginalized women's access to health care: a feminist narrative analysis.

In the emergent restructuring of health care, it is imperative to examine marginalized consumers' experiences with access to care so that the process addresses not merely the issue of rising health care costs, but also begins to meet the actual health care needs of diverse communities. This report of a feminist narrative study of access for low-income lesbians from several ethnic/racial groups provides critique of the US health care system from the standpoint of consumers and offers a transformative vision of what a just, effective, and compassionate system might encompass. By generating compelling evidence about the access afforded in current models of health care delivery, this study serves as an example of the significance of its methodological innovations, sampling from the margins, and narrative design.

Adult↗
PubMed2004

Flow cytometric analysis of kinase signaling cascades.

Flow cytometry offers the capability to assess the heterogeneity of cellular subsets that exist in complex populations, such as peripheral blood, based on immunophenotypes. We describe methodologies to measure phospho-epitopes in single cells as determinants of intracellular kinase activity. Multiparametric staining, using both surface and intracellular stains, allows for the study of discrete biochemical events in readily discernible lymphocyte subsets. As such, the usage of multiparameter flow cytometry to obtain proteomic information provides several major advantages: (1) the ability to perform multiparametric experiments to identify distinct signaling profiles in defined lymphocyte populations, (2) simultaneous correlation of multiple active kinases involved in signaling cascades, (3) profiling of active kinase states to identify signaling signatures of interest rapidly, and (4) biochemical access to rare cell subsets such as those from clinically derived samples or populations that comprise too few in numbers for conventional biochemical analysis.

Animals↗
PubMed1992

Method slurring: the grounded theory/phenomenology example.

Increasingly, qualitative research methods are being embraced by nurse researchers because these approaches allow exploration of human experience. Failure to explicate qualitative methodologies is resulting in a body of nursing research that is either mislabelled or is classified broadly as qualitative and subject to charges that qualitative research lacks rigour. In this paper, the authors discuss the importance of specificity in methodology and distinguish between phenomenology and grounded theory, two frequently misused terms in the description of qualitative methodology.

Data Collection↗
PubMed2000

Tell me, what did you see? The stimulus on computers.

Most psychology experiments start with a stimulus, and, for an increasing number of studies, the stimulus is presented on a computer monitor. Usually, that monitor is a CRT, although other technologies are becoming available. The monitor is a sampling device; the sampling occurs in four dimensions: spatial, temporal, luminance, and chromatic. This paper reviews some of the important issues in each of these sampling dimensions and gives some recommendations for how to use the monitor effectively to present the stimulus. In general, the position is taken that to understand what the stimulus actually is requires a clear specification of the physical properties of the stimulus, since the actual experience of the stimulus is determined both by the physical variables and by the psychophysical variables of how the stimulus is handled by our sensory systems.

Behavioral Sciences↗