Development of a Primary Human Cell Model for the Study of Human Cytomegalovirus Replication and Spread within Salivary Epithelium.

Various aspects of human cytomegalovirus (HCMV) pathogenesis, including its ability to replicate in specific cells and tissues and the mechanism(s) of horizontal transmission, are not well understood, predominantly because of the strict species specificity exhibited by HCMV. Murine CMV (MCMV), which contains numerous gene segments highly similar to those of HCMV, has been useful for modeling some aspects of CMV pathogenesis; however, it remains essential to build relevant human cell-based systems to investigate how the HCMV counterparts function. The salivary gland epithelium is a site of persistence for both human and murine cytomegaloviruses, and salivary secretions appear to play an important role in horizontal transmission. Therefore, it is important to understand how HCMV is replicating within the glandular epithelial cells so that it might be possible to therapeutically prevent transmission. In the present study, we describe the development of a salivary epithelial model derived from primary human "salispheres." Initial infection of these primary salivary cells with HCMV occurs in a manner similar to that reported for established epithelial lines, in that gH/gL/UL128/UL130/UL131A (pentamer)-positive strains can infect and replicate, while laboratory-adapted pentamer-null strains do not. However, while HCMV enters the lytic phase and produces virus in salivary epithelial cells, it fails to exhibit robust spread throughout the culture and persists in a low percentage of salivary cells. The present study demonstrates the utility of these primary tissue-derived cells for studying HCMV replication in salivary epithelial cells in vitroIMPORTANCE Human cytomegalovirus (HCMV) infects the majority of the world's population, and although it typically establishes a quiescent infection with little to no disease in most individuals, the virus is responsible for a variety of devastating sequelae in immunocompromised adults and in developing fetuses. Therefore, identifying the viral properties essential for replication, spread, and horizontal transmission is an important area of medical science. Our studies use novel human salivary gland-derived cellular models to investigate the molecular details by which HCMV replicates in salivary epithelial cells and provide insight into the mechanisms by which the virus persists in the salivary epithelium, where it gains access to fluids centrally important for horizontal transmission.

A proof-of-concept study of the "Employment and Arthritis: Making It Work" program.

OBJECTIVE: Work disability is a common outcome of inflammatory arthritis (IA), yet few services address employment. We conducted a proof-of-concept study of the "Employment and Arthritis: Making It Work" self-management program aimed at preventing work disability and maintaining at-work productivity in employed people with IA. METHODS: The program was developed using the precede-proceed model and self-management concepts. Program goals included modifying risk factors for work disability and enhancing self-management of work problems due to IA, as identified in initial focus groups. The program included a self-learning manual, 5 group sessions, and individual visits with an occupational therapist for an ergonomic assessment and a vocational rehabilitation counselor. It was pilot tested in 2 groups (n = 19) and evaluated over 12 months of followup. RESULTS: Participants consisted of 19 employed women with IA. Process evaluation demonstrated feasibility and excellent attendance and use of the self-learning manual. By 1 year, 80% reported increased confidence in requesting job accommodations, 74% had requested an accommodation, and 71% of requested accommodations were implemented. The occupational therapist and vocational rehabilitation counselor visits resulted in recommendations for change in 100% and 74% of participants, respectively, with implementation of some recommended changes in 89% and 63%, respectively. Improvements were observed in self-confidence in managing problems at work, fatigue interference with work, measures of limitations, and at-work productivity. CONCLUSION: We developed a novel intervention to prevent work disability in patients with IA, combining self-management group sessions and professional assessments aimed at job retention, which resulted in people making changes to adapt their work to their arthritis, and improved fatigue, self-efficacy, and at-work productivity.

Seeking Mind, Body and Spirit Healing-Why Some Men with Prostate Cancer Choose CAM (Complementary and Alternative Medicine) over Conventional Cancer Treatments.

UNLABELLED: Little is known about men with prostate cancer who decline conventional treatment and use only complementary and alternative medicine (CAM). OBJECTIVES: To 1) explore why men decline conventional prostate cancer treatment and use CAM 2) understand the role of holistic healing in their care, and 3) document their recommendations for health care providers. METHODS: Semi-structured interviews and follow-up focus groups. SAMPLE: Twenty-nine men diagnosed with prostate cancer who declined all recommended conventional treatments and used CAM. RESULTS: Based on strong beliefs about healing, study participants took control by researching the risks of delaying or declining conventional treatment while using CAM as a first option. Most perceived conventional treatment to have a negative impact on quality of life. Participants sought healing in a broader mind, body, spirit context, developing individualized CAM approaches consistent with their beliefs about the causes of cancer. Most made significant lifestyle changes to improve their health. Spirituality was central to healing for one-third of the sample. Participants recommended a larger role for integrated cancer care. CONCLUSION: Men who decline conventional prostate cancer treatment and use CAM only may benefit from a whole person approach to care where physicians support them to play an active role in healing while carefully monitoring their disease status.

Problems faced at work due to inflammatory arthritis: new insights gained from understanding patients' perspective.

OBJECTIVE: A qualitative study was conducted to better understand patients' perspective on their experience at work in relation to their inflammatory arthritis (IA). Objectives were to identify the problems and barriers to employment that persons with IA face at work because of arthritis, understand why these issues are problematic, and identify strategies helpful for maintaining employment. METHODS: Five focus groups were conducted with 36 employed adults with IA (75% rheumatoid arthritis) recruited from rheumatology practices and outpatient arthritis treatment programs. Script design used brainstorming techniques to identify problems and helpful strategies, and root cause analysis to capture in-depth information about underlying causes of problems. Descriptive qualitative analysis of transcripts was performed by 2 researchers independently to identify problems and organize them into topics and broad categories. RESULTS: Problems clustered around 4 categories: arthritis symptoms, working conditions, interpersonal difficulties at work, and emotional challenges. New insights gained included identifying fatigue as the aspect of IA most limiting employment; challenges posed by invisibility, fluctuation, and unpredictability of arthritis; complexity of interpersonal relationships at work; reluctance to disclose or draw attention to arthritis; numerous barriers to using available supports and requesting job accommodations, including fear of disclosure and concern it could be perceived by coworkers as favoritism; loss of self-efficacy at work; and many emotional challenges. CONCLUSION: This research identified new issues that are meaningful to individuals working with arthritis and that deserve greater attention by professionals counseling people on employment, in intervention efforts to help maintain employment, and in arthritis employment studies.

Galanin modulates cholinergic neurotransmission in the heart.

The role of galanin (Gal) in the modulation of cholinergic neurotransmission in the heart in wild-type (129 SvJ), and GALR1 knockout mice has been studied. The mice were anaesthetised and ventilated. Blood pressure (BP) and the increase in pulse interval evoked by stimulation of the vagus nerve (deltaPI) were recorded. Resting BP and PI were not different in control and GALR1-KO mice. In control mice an intravenous, bolus injection of Gal (0.8-13 nmol/kg; n = 4-6) attenuated the deltaPI, dose dependently from 33 +/- 7% to 78 +/- 9.5%. In GALR1-KO mice, Gal (0.8-13 nmol/kg) did not attenuate deltaPI at any dose (n = 3-4). In control mice intravenous, bolus injection of neuropeptide Y (NPY; 0.5-10 nmol/kg, n = 5-7) attenuated the deltaPI by 13 +/- 10% to 67 +/- 7% with a half time to recovery of 0.5-5 +/- 1 min. In control mice, following activation of the cardiac sympathetic nerve (10 Hz for 2 min; n = 3) the deltaPI was attenuated by 92 +/- 2% with a half time to recovery of 7 +/- 1 min. In control mice in the presence of the beta-adrenoceptor antagonist propranolol (1 mg/kg), and 1 micromol/kg BIIE0426 (an NPY Y2 receptor antagonist) the deltaPI was 57+/-3% with a half time to recovery of 2.5+/-0.5 min. In GALR1-KO mice, in the presence of propranolol and BIIE0426 there was no inhibition of deltaPI. In mice, it is proposed that both Gal and NPY contribute to the prolonged attenuation of parasympathetic slowing of the heart following activation of the cardiac sympathetic nerve.

Toward a patient-centered approach: incorporating principles of participatory action research into clinical studies.

UNLABELLED: Despite the shift to patient-centered care in recent years, many clinical studies continue to reinforce the traditional researcher/subject relationship. In contrast, action research engages study participants in a collaborative relationship with researchers. OBJECTIVES: To review the benefits of adding a participatory component to an existing study with respect to (1) engaging participants in the research process to clarify and validate qualitative findings, (2) engaging participants in the change process to develop potential solutions for improving integrative cancer services, and (3) giving voice to the concerns of patients using complementary and alternative medicine. METHOD: Focus groups were used to clarify concepts arising from patient interviews and to provide a forum for participants to develop recommendations and facilitate dissemination. CONCLUSION: Our approach empowered patients by involving them in the research and in developing solutions for how health care providers, policy makers, and researchers can enhance an integrative approach to cancer care.

Galanin and neuropeptide Y reduce cholinergic transmission in the heart of the anaesthetised mouse.

(1) This study investigated the effects of galanin (GAL) on inhibition of cholinergic (vagal) activity in the mouse heart using control galanin knockout (GAL-KO) and GAL-1R receptor knockout (GAL-1R-KO) mice. (2) In pentobarbitone anaesthetised mice, supramaximal stimulation every 30 s of the vagus nerve innervating the heart, increased pulse interval (PI) by approximately 50 ms or decreased heart rate by approximately 100 beats min-1. This response was attenuated by intravenous administration of GAL (dose ranged from 0.8 to 13 nmol kg-1) in a dose-dependent manner. (3) In GAL-KO mice, the magnitude of inhibition of the increase in PI (DeltaPI) following a bolus dose of GAL was not different from the DeltaPI in control mice, and neuropeptide Y (NPY), previously shown to attenuate vagal inhibitory activity in mice, evoked a comparative inhibition of DeltaPI in GAL-KO mice. (4) In GAL-1R-KO mice, an intravenous, bolus injection of GAL had no inhibitory effect on vagal activity. (5) In control mice, stimulation of the sympathetic nerve at 25 V, 10 Hz for 2 min in the presence of propranolol evoked a long-lasting attenuation of DeltaPI. The inhibitory effect on DeltaPI was reduced in the presence of the NPY Y2 antagonist, BIIE0246. (6) In GAL-1R-KO mice, stimulation of the sympathetic nerve in the presence of propranolol evoked an attenuation of DeltaPI not significantly different from the response in control mice in the presence of BIIE0246. Following administration of BIIE0246 in GAL-1R-KO mice, the inhibition of DeltaPI that followed stimulation of the sympathetic nerve was abolished. (7) These findings support the view that the nerve terminals of parasympathetic neurons in the mouse heart possess both GAL-1R and NPY Y2 receptors which, when activated, reduce acetylcholine release.

Decision-making control: why men decline treatment for prostate cancer.

UNLABELLED: The purpose of this study was to conduct a qualitative analysis of decision-making control by men with prostate cancer who refuse conventional cancer therapies. The transcripts for 8 prostate cancer patients from a larger qualitative study were analyzed separately to explore in depth the factors related to decision-making control. RESULTS: Most men were newly diagnosed when they made the decision to forgo conventional cancer treatment in favor of alternative approaches. Five areas were identified in which patients took control over the treatment process. These include control over (1) the timing of treatment, (2) information about conventional treatment and risk assessment, (3) designing an alternative treatment plan, (4) coordination of cancer care, and (5) monitoring and evaluation of disease progression. Clinicians can support patients who delay or forgo treatment for prostate cancer by helping them maintain a sense of control over the treatment process. This can be achieved by supporting patients' efforts to integrate complementary therapies into their cancer care, by addressing fears related to treatment early in the decision-making process, and by encouraging open communication about the reasons for seeking alternatives to conventional treatment. Findings from this study need to be evaluated in a larger, quantitative study.

Cardiac function in neuropeptide Y Y4 receptor-knockout mice.

Autonomic control of cardiovascular function in neuropeptide Y (NPY) Y4 receptor-knockout mice was investigated using pancreatic polypeptide (PP), NPY and specific agonists and antagonists for other NPY receptors well characterised in cardiovascular function. Y4 receptor-knockout mice, anaesthetised with sodium pentobarbitone, displayed slower heart rate, indicated by a higher pulse interval and lower blood pressure compared to control mice. After vagus nerves were cut heart rate increased but was still significantly slower than in control mice. PP had no effect on blood pressure or cardiac vagal activity in either group of mice, which was consistent with earlier studies in other species. Injection of NPY evoked an increase in blood pressure but the response was significantly reduced in Y4 receptor-knockout mice compared to the controls. The reduction in pressor activity was not Y1 mediated as the selective Y1 antagonist, BIBP 3226, was effective in blocking NPY pressor activity in knockout mice. In addition, cardiac vagal inhibitory activity evoked by low doses of NPY was also reduced when compared to control responses. As N-acetyl [Leu(28, 31)] NPY 24-36 inhibited vagal activity dose dependently in both groups of mice with no difference in response at any dose, it is unlikely that this effect also is receptor mediated. We propose that the reduced vasoconstrictor and vagal inhibitory activity evoked by NPY in Y4 receptor-knockout mice is due to a lack of adrenergic tone bought about by a proposed reduction in sympathetic activity, possibly resulting from altered NPY activity secondarily affecting adrenergic transmission. We conclude that Y4 receptor deletion disrupts autonomic balance within the cardiovascular system.

Factors in making the decision to forgo conventional cancer treatment.

PURPOSE: The purpose of this study was to explore why and how patients with cancer decide to forgo conventional cancer treatments in favor of alternative treatments and which factors influence such decisions. DESCRIPTION OF STUDY: Due to the exploratory nature of the study, this was a qualitative study using focus groups and in-depth interviews in a convenience sample of patients. All patients had received diagnoses of cancer and had refused one or more conventional treatments offered to them by their cancer healthcare professionals. RESULTS: Thirty-one persons with cancer, widely varying in age and tumor sites, volunteered to take part in the study. Of these, 12 refused all conventional treatment, 13 refused most or some of the treatments recommended, and 6 discontinued conventional treatment. The decision-making model, which emerged from the data, identifies several groups of variables. These include factors that predispose participants to the decision to forgo conventional treatment(s), such as having a close relative or friend who has died from cancer when receiving conventional treatment; experiences around the diagnosis; and factors relevant after the diagnosis, such as beliefs, need for control, side effects of conventional cancer treatment, and communication with physicians. Last, perceived outcomes of the decision proved to be an important theme in the focus groups and interviews. CLINICAL IMPLICATIONS: Patients with cancer may benefit from counseling to help them explore the difference between their diagnosis and treatment plan and those of family members or friends who died of cancer while receiving conventional treatment. Counseling also may be helpful in resolving emotional issues underlying the decision to forgo treatment. Last, patients should have access to healthcare professionals, including physicians and counselors, who would assist them with their decision making without judging or intimidating them.

Stabilization of the helical structure of Y2-selective analogues of neuropeptide Y by lactam bridges.

The importance of helical structure in an analogue of NPY selective for the Y2 receptor, Ac[Leu28,31]NPY24-36, has been investigated by introducing a lactam bridge between positions 28 and 32. The resulting analogue, Ac-cyclo28/32[Ala24,Lys28,Leu31,Glu32]NPY24-36, is a potent Y2-selective agonist. Structural analysis by NMR shows that this analogue forms a helical structure in a 40% trifluoroethanol/water mixture, whereas in water only the region around the lactam bridge (Lys28-Glu32) adopts helical-like structure, with both N- and C-termini being poorly defined. The observation of well-defined helical structure in aqueous TFE contrasts with that reported for a similar analogue, Ac-cyclo28/32[Lys28,Glu32]NPY25-36 (Rist et al. FEBS Lett. 1996, 394, 169-173), which consisted of a hairpin-like structure that brought the N- and C-termini into proximity. We have therefore determined the structures of this analogue, as well as those of Ac-cyclo28/32[Ala24,Lys28,Leu31,Glu32]NPY24-36 and Ac-cyclo28/32[Ala24,Lys28,Glu32]NPY24-36, under identical solution conditions (30% TFE/H2O mixture at 308 K) and find essentially the same helical structure in all three peptides. These findings support the proposal that these Y2-selective analogues adopt a helical structure when bound to the Y2 receptor.

Role of neuropeptide Y Y(2) receptors in modulation of cardiac parasympathetic neurotransmission.

The aim of the study was to clarify the role of the Y(2) receptor in regulation of vagal control of the heart, using Y(2)((-/-)) receptor-knockout mice. Adult Y(2)((+/+),(-/-)) mice (50% C57BL/6-50% 129/SvJ background) were anaesthetised and artificially ventilated. Arterial blood pressure and pulse interval was recorded and both vagus nerves were cut. The cardiac end of the right vagus nerve was stimulated supra-maximally every 30 s (7 V, 2-2.5 Hz, 5 s). Neuropeptide Y (NPY) and a Y(2) receptor agonist, N-acetyl [Leu(28, 31)]NPY 24-36, were injected intravenously in both groups of mice. N-acetyl [Leu(28, 31)] NPY 24-36 was also administered to control mice in the presence of a Y(2) receptor antagonist, BIIE0246. Stimulation of the vagus nerve increased pulse interval (PI) by approximately 100 ms. NPY and N-acetyl [Leu(28, 31)] NPY 24-36 attenuated the increase in PI evoked by vagal stimulation in control mice only. The attenuation was reduced in the presence of BIIE0246. The results presented here show in Y(2)((-/-)) receptor-knockout mice that NPY and N-acetyl [Leu(28, 31)] NPY 24-36 have no effect on PI evoked by vagal stimulation. These findings demonstrate that NPY attenuates parasympathetic activity to the heart via the Y(2) receptor.