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Additional file1. Additional figures. Fig. S1 Galectin-3 is upregulated in the BAL fluid of mice following IAV infection. Mice were intratracheally inoculated with IAV (105 PFU) at day 0, and the BAL fluid was collected at different time points. a Immunoblotting of galectin-3 in the BAL fluid of individual mice. b The intensity of the 30-kDa band corresponding to galectin-3 was determined by densitometric analysis, and relative expression levels of galectin-3 at different time points after viral infection were compared, where the ratio of day 0 was arbitrarily set to 1. Values shown are mean ± SD (n = 5). Fig. S2 Recombinant mouse galectin-3 proteins induce macrophage migration at low concentration as well as binds to IAV and inhibits viral hemagglutination activity at high concentrations. a, b Microscopic images (a) and quantification of migratory cells (b) in the Boyden chamber assay. RAW 264.7 cells and various concentrations of recombinant galectin-3 proteins were applied to the upper and lower chambers, respectively. After 6 h, cells that migrated through the membrane to the lower surface were stained and quantified. The number of migratory cells was the average of the cells counted in three randomly selected fields in each well (n = 3). c Binding of galectins to IAV. Serial two-fold dilutions of galectin-3 or galectin-1, ranging from 100 μg/well to 6.25 μg/well, were applied to 96-well plates coated with IAV (4 HAU/well). The bound galectin-3 and galectin-1 proteins were detected by ELISA with anti-galectin-3 and anti-galectin-1 antibodies, respectively. Note that galectin-3 bound to IAV much weaker than galectin-1. d Hemagglutination inhibition activity of galectin-3. IAV (2 HAU) was incubated with various concentrations of galectin-3 for 60 min, followed by addition of human erythrocyte suspension. After an additional 60-min incubation, the presence or inhibition of hemagglutination was recorded. Fig. S3 Treatment with recombinant mouse galectin-3 proteins does not impact body weight and survival of IAV-infected mice. Groups of C57BL/6 mice were intrantracheally inoculated with IAV (106 PFU) at day 0 and treated with mouse galectin-3 proteins (50 μg) or BSA at days 2, 4, and 5 p.i. via the same route. a Changes in body weights from day 0 through day 4 while all mice were still alive. Body weights were recorded and expressed as the percentage of pre-infection (day 0) body weight. b Kaplan-Meier survival curves. Values shown are mean ± SD (n = 5). Fig. S4 Exogenous galectin-3 affects the nucleocytoplasmic distribution of the vRNP complex in IAV-infected A549 cells. a, b A549 cells treated with recombinant human galectin-3 proteins (2 μg/ml) or untreated cells were infected with IAV at an MOI of 5 for 60 min, fixed, and then permeabilized at 2 h, 3.5 h, 4.5 h, and 6 h p.i. for immunofluorescence staining with the anti-NP antibody. Nuclei were stained with DAPI. Localization of the viral NP indicative of the vRNP complex (a) and quantitative analysis of nucleocytoplasmic distribution of the NP at 6 h p.i. (n = 4) (b, c). NP-positive (green staining) cells were examined for their nuclear/cytoplasmic localization at 6 h p.i. Representative images (original magnification × 200, scale bar = 50 μm) (a) and percentages of the NP observed only in the nucleus (b) and in both the nucleus and cytoplasm (c) are shown.

Additional file 1. Supplemental Figure 1. The effect of IAV NS1 1-124 on blocking RIG-I signaling to IFN-β activation. Supplemental Figure 2. IAV NS1 is co-localized with TRAF3 in mammalian cells. Supplemental Figure 3. IAV NS1 uses its ED-C to block the K63-linked ubiquitination of TRAF3 during RIG-Isignaling. Supplemental Figure 4. IAV NS1 E152/E153 residues are not essential for blocking the NF-κB promoter activity during RIG-I, TLR3 or TLR7 signaling. Supplemental Figure 5. NS1 E152/E153 residues confer the in vivo pathogenicity of IAV infection. Supplemental Table 1. Primers used for RT-PCR, cloning and mutagenesis. Supplemental Table 2. Antibodies.

This study was motivated by the researcher’s experience of working in end-of-life care and by the literature review which revealed a gap in the knowledge and understanding of the role of religious methods as complementary approaches in managing the experience of living with a life-limiting illness in Taiwan. Trans-cultural issues are extremely important to end-of-life care. In Taiwan, patients approaching death have used religious methods as complementary approaches to manage the experience of living with a life-limiting illness, and religious belief systems shape patients’ understandings of what is happening. Current literature coupled with the experience of palliative care personnel identified that some patients with religious persuasions were refusing western medical treatments when they recognised that they were in the end stage of disease because they believed that these treatments could not control death and rebirth. However, few studies have discussed this experience and its meaning. Buddhist chaplains, as providers of supportive palliative care services through therapeutic care, have presented their understanding of the way that people move towards death and dying in Buddhist temples, universities, and in public speeches, but not often in hospitals. Buddhist chaplains’ life experience and interpretations influence the thinking processes and decision-making of many of those they come in contact with, especially those who share the Buddhist faith. However, few studies have demonstrated the way in which patients have made use of religious methods as complementary approaches from the perspective of hospice palliative Buddhist chaplains. The perspectives and work of hospice palliative Buddhist chaplains regarding “hospice palliative care” and patients’ use of religious methods as complementary approaches in end-of-life care in Taiwan were explored. The research questions were: (1) How do the Buddhist chaplains define “hospice palliative care”? (2) How do Buddhist chaplains use Buddhist religious methods as complementary approaches in clinical end-of-life care? (3) What are the experiences of Buddhist chaplains regarding the patients’ use of Buddhist religious methods as complementary approaches in clinical end-of-life care? (4) What are the opinions of Buddhist chaplains regarding patients’ use of Buddhist religious methods as complementary approaches in clinical end-of-life care? Charmaz’s (2006) constructivist grounded theory method was adopted. Data collection used triangulation and included demographic questionnaires, semi-structured face-to-face interviews, field notes, and written memos. Purposive sampling was used to recruit participants with rich working experiences in clinical end-of-life care. Twenty female and two male Buddhist chaplains aged between 33 and 67 years old participated. Charmaz’s (2006) constructivist grounded theory, which included comparative method, and three analytical phases (initial coding, focused coding and theoretical coding) informed the data analysis. The findings demonstrate that Buddhist concepts of death, the process of dying, and the ethics and tools of the Buddhist religion formed the basis of the practice of the chaplains who regarded compassionate care and Mahayana Buddhism as the main content of Buddha’s teachings. All participants used aspects of Buddhist philosophy to define “hospice palliative care”. The final theoretical framework emerged from the data to provide a structure to interpret “the dynamic process of compassionate care”. Compassionate care is a multifaceted, dynamic phenomenon practised by the chaplains. Mahayana Buddhism provides the specific tools through which they interacted with patients creating a sacred relationship that allowed patients to understand their context and cope with their end-of-life experiences. The thesis concluded that Buddhist chaplains’ understanding of compassionate care was influenced by their educational background, hospice training courses, and Buddha’s teachings to enable them to play important roles in end-of-life care in Taiwan. Recommendations are made for future studies to test the theoretical framework regarding “the dynamic process of compassionate care” with different professional staff such as nurses, psychologists and mental health physicians. The findings are also relevant for future government policy concerning the financial cost of end-of-life care which is currently provided by Buddhist chaplains from a Charity rather than by Taiwanese National Health Insurance. Finally it recommends that the findings inform the future education of medical and nursing students and staff in hospice end-of-life care in Taiwan.

Validerat; 20101217 (root)