Purification of cardiac myocytes from human heart biopsies for gene expression analysis.

The collection of gene expression data from human heart biopsies is important for understanding the cellular mechanisms of arrhythmias and diseases such as cardiac hypertrophy and heart failure. Many clinical and basic research laboratories conduct gene expression analysis using RNA from whole cardiac biopsies. This allows for the analysis of global changes in gene expression in areas of the heart, while eliminating the need for more complex and technically difficult single-cell isolation procedures (such as flow cytometry, laser capture microdissection, etc.) that require expensive equipment and specialized training. The abundance of fibroblasts and other cell types in whole biopsies, however, can complicate gene expression analysis and the interpretation of results. Therefore, we have designed a technique to quickly and easily purify cardiac myocytes from whole cardiac biopsies for RNA extraction. Human heart tissue samples were collected, and our purification method was compared with the standard nonpurification method. Cell imaging using acridine orange staining of the purified sample demonstrated that >98% of total RNA was contained within identifiable cardiac myocytes. Real-time RT-PCR was performed comparing nonpurified and purified samples for the expression of troponin T (myocyte marker), vimentin (fibroblast marker), and alpha-smooth muscle actin (smooth muscle marker). Troponin T expression was significantly increased, and vimentin and alpha-smooth muscle actin were significantly decreased in the purified sample (n = 8; P < 0.05). Extracted RNA was analyzed during each step of the purification, and no significant degradation occurred. These results demonstrate that this isolation method yields a more purified cardiac myocyte RNA sample suitable for downstream applications, such as real-time RT-PCR, and allows for more accurate gene expression changes in cardiac myocytes from heart biopsies.

Identification of predicted human outer dynein arm genes: candidates for primary ciliary dyskinesia genes.

BACKGROUND: Primary ciliary dyskinesia (PCD) is a severe inherited disorder characterised by chronic respiratory disease, male infertility, and, in approximately 50% of affected individuals, a left-right asymmetry defect called situs inversus. PCD is caused by defects in substructures of the ciliary and flagellar axoneme, most commonly loss of the outer dynein arms. Although PCD is believed to involve mutations in many genes, only three have been identified. METHODS: To facilitate discovery of new PCD genes, we have used database searching and analysis to systematically identify the human homologues of proteins associated with the Chlamydomonas reinhardtii outer dynein arm, the best characterised outer arm of any species. RESULTS: We find that 12 out of 14 known Chlamydomonas outer arm subunits have one or more likely orthologues in humans. The results predict a total of 24 human genes likely to encode outer dynein arm subunits and associated proteins possibly necessary for outer arm assembly, plus 12 additional closely related human genes likely to encode inner dynein arm subunits. CONCLUSION: These genes, which have been located on the human chromosomes for easy comparison with known or suspected PCD loci, are excellent candidates for screening for disease-causing mutations in PCD patients with outer and/or inner dynein arm defects.

Treatments to promote colonic hydration: enteral fluid therapy versus intravenous fluid therapy and magnesium sulphate.

Although large intestine impactions are commonly treated with i.v. fluids combined with the osmotic laxative MgSO4, enteral fluids are less expensive and also appear to be efficacious for impactions. Therefore, this study was conducted to compare the systemic and gastrointestinal effects of enteral fluids with the changes produced by i.v. fluids combined with MgSO4. Four horses with a fistula in the right dorsal colon alternately received both treatments in 2 periods one week apart. Sixty litres of fluids were administered continuously (10 l/h) through a venous catheter or a nasogastric tube. Magnesium sulphate (1 g/kg bwt) was administered via nasogastric tube before i.v. fluid therapy. Two horses had mild abdominal discomfort at the end of enteral fluid therapy. Pollakiuria, hypostenuria, increased bodyweight, increased faecal and ingesta hydration, and decreased PCV, plasma protein and plasma magnesium were produced by both treatments. Abdominal distention and more pronounced changes in bodyweight and ingesta hydration were seen with enteral fluids. Intravenous fluids plus MgSO4 produced hypocalcaemia and more pronounced changes in plasma protein. These results indicate that enteral fluid therapy is more effective in promoting ingesta hydration and produces less pronounced systemic effects than i.v. fluid therapy plus MgSO4.

Iron metabolism in mammalian cells.

Most living things require iron to exist. Iron has many functions within cells but is rarely found unbound because of its propensity to catalyze the formation of toxic free radicals. Thus the regulation of iron requirements by cells and the acquisition and uptake of iron into tissues in multicellular organisms is tightly regulated. In humans, understanding iron transport and utility has recently been advanced by a "great conjunction" of molecular genetics in simple organisms, identifying genes involved in genetic diseases of metal metabolism and by the application of traditional cell physiology approaches. We are now able to approach a rudimentary understanding of the "iron cycle" within mammals. In the future, this information will be applied toward modulating the outcome of therapies designed to overcome diseases involving metals.

Lues and lupus: syphilis mimicking systemic lupus erythematosus (SLE).

Malar rash and photosensitivity are common findings in systemic lupus erythematosus (SLE). However, a number of inflammatory, dermatologic, infectious or drug-induced conditions may mimic cutaneous findings of SLE. These typically include seborrheic dermatitis, contact dermatitis, rosacea, polymorphous light eruption, syphilis and dermatomyositis sine myositis. Herein we describe a patient with fever, malar rash, alopecia, photosensitivity, arthralgias and lymphadenopathy who was subsequently diagnosed with secondary syphilis. In this case report we review clinical and histopathological findings in the differential diagnosis of malar rash and photosensitivity and discuss the overlapping features of syphilis and SLE.

Psychometric testing of fatigue instruments for use with cancer patients.

BACKGROUND: Cancer treatment-related fatigue (CRF) is a common side effect of cancer treatment. A problem identified in most reviews of CRF is lack of sound approaches to measurement that are congruent with the conceptualization of CRF as a self-perceived state. The diversity of instruments available to measure fatigue and the lack of comprehensive testing of several promising instruments with cancer patients undergoing treatment provided the rationale for this study. The purpose of this article is to report the results of psychometric testing of several fatigue instruments in patients undergoing cancer treatment. OBJECTIVES: The aims of this study were to determine the reliability, validity, and responsiveness of each instrument and to determine the ability of each instrument to capture CRF. METHODS: Existing fatigue instruments with published psychometric information that indicated suitability for further testing were selected and included the Profile of Mood States Short Form fatigue subscale (F_POMS-sf), Multidimensional Assessment of Fatigue (MAF), Lee Fatigue Scale (LFS), and the Multidimensional Fatigue Inventory (MFI). Data were collected at a university-based clinical cancer center and a freestanding comprehensive cancer center. Subjects completed all study instruments, which were presented in random order, at a time when CRF was expected to be high and again when it was expected to be low. A subset of subjects completed the instruments within 48 hours of one of the data collection points when CRF was expected to be relatively unchanged to provide stability data. RESULTS: Reliability estimates using Cronbach's alpha indicated that all instruments examined had good internal consistency. Test-retest correlations showed good stability for total scores on all the instruments, but some subscales of the LFS and MFI had marginal stability. Factor analysis of all instruments indicated that only the LFS and the F_POMS-sf fully supported their construct validity. All of the instruments showed responsiveness to changes in CRF related to treatment. CONCLUSIONS: The results of the study provide researchers and clinicians with detailed comparisons of the performance of established fatigue measures in cancer patients undergoing treatment to use when selecting measures of CRF.

Does emotional expression make a difference in reactions to breast cancer?

PURPOSE/OBJECTIVES: To examine the feasibility of using an emotional expression intervention with patients with cancer and test the hypothesis that emotional expression improves psychosocial adjustment. DESIGN: Sequentially randomized pretest/post-test design with repeated measures. SETTING: Two radiation therapy (RT) facilities. SAMPLE: Women completing RT for stage I or II breast cancer, who spoke and read English, were independent in self-care, and provided written consent. Subjects (N = 44) were middle-aged (mean = 53.6 years), Caucasian, married, and well educated. METHODS: Following a baseline interview, subjects were sequentially randomized to an attentional control group, a single dose, or a three-dose emotional expression writing group. Interventions were administered at the time of completion of RT. Follow-up telephone interviews were completed at 1, 4-6, 16, and 28 weeks post-RT. MAIN RESEARCH VARIABLES: Positive and negative affect, intrusiveness of thoughts, use of avoidant coping, side effect severity, trait negative affectivity, content of written essay, and themes derived from content analysis. FINDINGS: A high level of acceptance and completion of emotional expression existed, but no effect of the intervention on psychosocial adjustment was evident. Process measures in the three-dose group changed as expected. No relationship existed between content changes and outcome measures. CONCLUSIONS: Emotional expression is feasible for patients with cancer, but the efficacy of the intervention in improving mood and decreasing cognitive intrusion and avoidance was not supported. Emotional expression processes were consistent with those seen in other samples and may influence outcomes that were not addressed in this study. IMPLICATIONS FOR NURSING PRACTICE: More extensive testing is needed, including additional outcome variables. Essays reveal concerns around communication, recurrence, and health behavior changes that should be considered in practice.

Self-care and breast cancer recovery.

PURPOSE: This study identified self-care behaviors used by women during and after radiation therapy for early-stage breast cancer. DESCRIPTION OF STUDY: Interviews with 23 women 7 months post-radiation therapy for breast cancer were analyzed for descriptions of self-care. The interviews were part of the final data collection in a longitudinal study investigating experiences post-treatment. RESULTS: Self-care behaviors reported most frequently were establishing a good support network, living life to the fullest despite illness, regulating feelings, managing stress, and taking responsibility for one's own health. Results suggest that breast cancer patients have many needs that continue to require attention post-radiation therapy, making self-care an important consideration throughout recovery. CLINICAL IMPLICATIONS: Quality of self-care may make a significant difference to women after treatment for breast cancer. Healthcare providers can promote self-care by encouraging patients to conserve energy, to engage in relationships, and to maintain a positive outlook and by suggesting activities that help patients cope, adapt, and manage symptoms during and after treatment.

Planning and conducting a multi-institutional project on fatigue.

PURPOSE/OBJECTIVES: To describe the process used in proposal development and study implementation for a complex multisite project on cancer treatment-related fatigue (CRF), identify strategies used to manage the project, and provide recommendations for teams planning multisite research. DATA SOURCES: Information derived from project team meeting records, correspondence, proposals, and personal recollection. DATA SYNTHESIS: The project was built on preexisting relationships among the three site investigators who then built a team including faculty, research coordinators, staff nurses, and students. Study sites had a range of organizational models, and the proposal was designed to capitalize on the organizational and resource strengths of each setting. Three team members drawn from outside oncology nursing provided expertise in measurement and experience with fatigue in other populations. Planning meetings were critical to the success of the project. Conference calls, fax technology, and electronic mail were used for communication. Flexibility was important in managing crises and shifting responsibility for specific components of the work. The team documented and evaluated the process used for multisite research, completed a major instrumentation study, and developed a cognitive-behavioral intervention for CRF. CONCLUSIONS: Accomplishments during the one-year planning grant exceeded initial expectations. The process of conducting multisite research is complex, especially when the starting point is a planning grant with specific research protocols to be developed and implemented over one year. Explicit planning for decision-making processes to be used throughout the project, acknowledging the differences among the study settings and planning the protocols to capitalize upon those differences, and recruiting a strong research team that included a member with planning grant and team-building expertise were essential elements for success. IMPLICATIONS FOR NURSING PRACTICE: Specific recommendations for others planning multisite research are related to team-building, team membership, communication, behavioral norms, role flexibility, resources, feedback, problem management, and shared recognition.

Basics of research (Part 12): Qualitative research.

Although both the assumptions and methods of qualitative and quantitative approaches in nursing research are different, both have the goal of furthering the scientific basis for practice. A variety of qualitative approaches are available, and which approach to use depends on the purpose of the research. In general, qualitative investigations address broad questions related to description, discovery, or theory building, and, as a consequence, the researcher is concerned with the entire context surrounding the phenomenon of interest rather than concentrating on specific variables thought to influence that phenomenon. The type of data collected and the methods of analysis differ, but qualitative research demands the same careful attention to selecting a design appropriate to answer the research question and the same assurance of rigor in conducting the research and interpreting the results as is required in quantitative studies. When these issues are thoroughly addressed, the clinician has a basis for judging both the accuracy and the applicability of qualitative research findings.

Adjustment of husbands and wives to breast cancer.

PURPOSE: The purpose of this descriptive study was to explore selected elements within the spousal relationship on fear of recurrence and emotional distress among women treated for breast cancer within the past 3 years and their husbands. DESCRIPTION OF STUDY: Fifty-eight married couples completed questionnaires designed to investigate the association between three relationship elements and adjustment (fear of recurrence and emotional distress) among husbands and wives. The relationship elements investigated were 1) adaptability and cohesion of the couple; 2) perceptions of support from the spouse; and 3) communication about the illness. The influence of the wife's age, treatment, and time since treatment also were considered. RESULTS: Both husbands and wives reported similar perceptions of dyadic functioning, level of support received from the spouse, and extent of communication about the illness. Interdependent elements within the couple relationship were associated with both fear of recurrence and emotional distress. Overall, the amount of communication about the illness explained the most variance in adjustment for both spouses, with communication increasing as fear of recurrence increased. There was some suggestion that current or past membership in a support group was related both to more communication by wives and to husbands' greater satisfaction with communication. For wives, age was the demographic variable most highly correlated with adjustment; the younger the wife, the greater was her fear of recurrence. Significant emotional distress was observed in a small subset of husbands. In addition, when either the husband or wife scored in the top 25% of the sample on fear of recurrence, the fear of recurrence score for the corresponding spouse was elevated in only two cases. This same pattern was true for emotional distress. CLINICAL IMPLICATIONS: This study adds to a growing body of information supporting the need to view women who have had breast cancer and their spouses as a unit. Four areas for heightened clinical assessment were noted; 1) greater fear of recurrence in younger women; 2) significant emotional distress in a small number of husbands; 3) the demonstration of compensation mechanisms by one spouse when the other is experiencing elevated fear of recurrence or emotional distress; and 4) an increased fear of recurrence in association with increased communication about the illness. Further investigation is necessary to determine whether communication between spouses is directed at problem solving or is an expression of emotional distress.

Concerns, affect, and cognitive disruption following completion of radiation treatment for localized breast or prostate cancer.

PURPOSE/OBJECTIVES: To describe concerns, cognitive disruption, and affect following the completion of radiation treatment (RT) for localized breast or prostate cancer. DESIGN: Cross-sectional, descriptive. SETTING: Two RT centers associated with a university hospital. SAMPLE: Patients who completed RT for prostate (n = 9) or breast cancer (n = 11). The mean age of the sample is 64 years. Ninety percent are Caucasian, 80% are married, 50% are of Latter-day Saint faith, and 50% are retired. All had early stage disease, with a mean of 20 months post-RT. METHODS: One-time interview using qualitative methods and the Life Orientation Test, Revised Impact of Events Scale, state scale of the Positive and Negative Affect Schedule, satisfaction items of the Sarason Social Support Questionnaire, and Side Effect Severity Checklist. MAIN RESEARCH VARIABLES: Frequency of intrusive thoughts or avoidant behavior, positive and negative affect, dispositional optimism, satisfaction with social support, severity of side effects, and themes extracted from semistructured interviews. FINDINGS: Most subjects had unexpected intrusive thoughts about cancer; more than one-third avoided reminders of cancer. The level of positive affect was high, as was optimism. The level of negative affect was low. Subjects' satisfaction level with social support was high. Fatigue was the most frequently reported continuing side effect. Relative optimism, acceptance, vigilance, and trust of healthcare providers were major themes. Avoidance, comparison, maintaining normalcy, and information seeking were common coping strategies. CONCLUSIONS: Subjects perceived themselves to be doing well. Fatigue was the most common long-term side effect. Subjects continued to seek information. Symptom vigilance and interpretation were important, as were having a positive outlook and maintaining normalcy. IMPLICATIONS FOR NURSING PRACTICE: Nurses must acknowledge the potential for concerns about symptoms and continuing side effects and what they mean for the patient. Clinicians need to provide accurate information about common post-treatment experiences. Clinicians also must remember that subjects have vivid recollections of both positive and negative interactions with healthcare providers.

Computer analysis of qualitative data: a comparison of three packages.

This article describes and compares three computer programs designed to assist in the analysis of narrative text. Ethnograph, GATOR, and Martin were compared in the context of a qualitative study of women and their husbands following mastectomy in order to (a) describe the fit between research purpose and program capabilities and (b) explore the influence, if any, of particular program features on research methods or analysis. The comparison suggests that methodological assistance provided by the programs involves differences in approach sufficient to make prior consideration of the fit between program capabilities and research purposes, methods, and style an important research decision. Descriptions of the programs and their application provide guidelines for comparison of these and other computer programs.

The short-term effects of a fire safety education program for the elderly.

The high risk of fire death and injury among elderly people is well documented. To be effective, fire safety education must reach older adults in the settings in which they reside: nursing homes and other long-term care institutions, board and care homes, and independent living facilities including the person's own home. Training must also be targeted at the people who are responsible for fire safety. In the case of the nursing home or board and care home, the responsible people are the staff and owners. In the case of the majority of older adults who live independently in their homes, it is either the individual or family members. These programs must also be comprehensive. A fire safety education curriculum was developed by a group of experts in a variety of related fields including fire safety, gerontology, health care industry, developmental disabilities, research, and instructional design. Older adults were included in each planning session. Based on that curriculum, workshops and workshop materials were developed for each of the three target populations: staff of health care facilities, staff and owners of board and care homes, and elderly people living independently in their homes. Materials included both print and audiovisuals. A pilot test of each workshop was conducted to test the short-term effects of the programs. Results indicated significant gains in knowledge for all groups and a significant improvement in positive attitudes toward fire safety for most participants. Measures of effects of the programs on intentions to change fire safety practices indicated a potential for change. Results also showed that the measured traits, knowledge of fire safety and attitudes toward fire safety, were relatively stable.