The IL-33:ST2 axis is unlikely to play a central fibrogenic role in idiopathic pulmonary fibrosis.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disease (ILD) with limited treatment options. Interleukin-33 (IL-33) is proposed to play a role in the development of IPF however the exclusive use of prophylactic dosing regimens means that the therapeutic benefit of targeting this cytokine in IPF is unclear. METHODS: IL-33 expression was assessed in ILD lung sections and human lung fibroblasts (HLFs) by immunohistochemistry and gene/protein expression and responses of HLFs to IL-33 stimulation measured by qPCR. In vivo, the fibrotic potential of IL-33:ST2 signalling was assessed using a murine model of bleomycin (BLM)-induced pulmonary fibrosis and therapeutic dosing with an ST2-Fc fusion protein. Lung and bronchoalveolar lavage fluid were collected for measurement of inflammatory and fibrotic endpoints. Human precision-cut lung slices (PCLS) were stimulated with transforming growth factor-ß (TGFß) or IL-33 and fibrotic readouts assessed. RESULTS: IL-33 was expressed by fibrotic fibroblasts in situ and was increased by TGFß treatment in vitro. IL-33 treatment of HLFs did not induce IL6, CXCL8, ACTA2 and COL1A1 mRNA expression with these cells found to lack the IL-33 receptor ST2. Similarly, IL-33 stimulation had no effect on ACTA2, COL1A1, FN1 and fibronectin expression by PCLS. Despite having effects on inflammation suggestive of target engagement, therapeutic dosing with the ST2-Fc fusion protein failed to reduce BLM-induced fibrosis measured by hydroxyproline content or Ashcroft score. CONCLUSIONS: Together these findings suggest the IL-33:ST2 axis does not play a central fibrogenic role in the lungs with therapeutic blockade of this pathway unlikely to surpass the current standard of care for IPF.

A national survey of oncology survivors examining nutrition attitudes, problems and behaviours, and access to dietetic care throughout the cancer journey.

BACKGROUND: Attitudes of cancer survivors to nutrition and nutrition care have rarely been captured. A better understanding of their needs based on a review of their experiences would give voice to this patient group (which has rarely been captured) and allow for better planning of nutritional care. AIMS: To conduct a national survey to determine: (1) survivors' experience in relation to nutrition and diet-related problems, (2) perceived importance of the role of nutrition to cancer survivors, (3) the experience of accessing dietetic support, (4) the sources where survivors get nutrition information, and (5) their use of alternative dietary strategies. METHODS: Survivors (any adult ever diagnosed with cancer) who had been diagnosed with or treated for cancer in Ireland within the past 5 years, were asked to complete a 25-item paper-based survey at one of 20 different hospital sites in Ireland. The survey was also hosted online on the websites of major cancer charities. Descriptive statistics were used to examine quantitative data. RESULTS: In total, 1073 valid responses were received (63% female, mean age 57 years (range 18-88)). Breast cancer was the most common (n = 362), followed by colorectal (n = 121). One third of respondents had metastatic disease. Diet-related problems were reported by 45%. Weight loss was experienced by 44% and amongst those, 42% reported they were 'unhappy or worried' by this, while 27% reportedbeing 'delighted/happy' with their weight loss. Muscle loss was noted by 52%, with 20% reporting they had noticed 'a lot' of muscle loss. Nutrition was rated as 'very/extremely' important to cancer care by 89% of respondents, yet 58% reported being asked about dietary issues by their medical team only 'sometimes', 'rarely' or 'never'. Only 39% had been assessed/treated by a registered dietitian (RD) and 74% rated their advice/care as 'very/extremely' helpful. Worryingly, 39% of survivors with involuntary weight loss, and 29% of survivors on a texture modified diet had not received nutritional care from an RD. Overall, 57% of those who did not see an RD said they wanted more dietetic support (access to a helpline/dietitian/additional reliable information). Of concern, 37% of survivors were following or had tried alternative, unproven dietary strategies (e.g. restrictive diets, herbal remedies, juicing or detoxes), and 32% reported avoiding specific foods, e.g. processed meat or dairy. A majority (56%) felt confused by the often conflicting nutrition information available in the media and offered by people around them. CONCLUSIONS: While nutrition is considered highly important by cancer survivors and a high proportion experience potentially serious diet-related problems including weight and muscle loss, fewer than half surveyed had access to a dietitian. Over a third had used at least one alternative dietary strategy, and over half felt confused about nutrition. Comprehensive nutritional screening and referral programmes to oncology dietitians need to be implemented in the ambulatory setting in order to identify and facilitate early management of the nutritional concerns of cancer survivors.

Dynamics of Colon Monocyte and Macrophage Activation During Colitis.

Background: Macrophages are pivotal in coordinating a range of important processes in the intestines, including controlling intracellular infections and limiting damaging inflammation against the microbiota. However, it is not clear how gut macrophages, relative to recruited blood monocytes and other myeloid cells, contribute to the intestinal inflammatory milieu, nor how macrophages and their monocyte precursors mediate recruitment of other immune cells to the inflamed intestine. Methods: Myeloid cell populations isolated from colonic inflammatory bowel disease (IBD) or murine dextran sulphate sodium (DSS) induced colitis were assessed using flow cytometry and compared to healthy controls. In addition, mRNA expression profiles in human and murine colon samples, and in macrophages and monocytes from healthy and inflamed murine colons, were analysed by quantitative PCR (qPCR) and mRNA microarray. Results: We show that the monocyte:macrophage balance is disrupted in colon inflammation to favour recruitment of CD14+HLA-DRInt cells in humans, and Ly6CHi monocytes in mice. In addition, we identify that murine blood monocytes receive systemic signals enabling increased release of IL-1ß prior to egress from the blood into the colon. Further, once within the colon and relative to other myeloid cells, monocytes represent the dominant local source of both IL-1ß and TNF. Finally, our data reveal that, independent of inflammation, murine colon macrophages act as a major source of Ccl7 and Ccl8 chemokines that trigger further recruitment of their pro-inflammatory monocyte precursors. Conclusions: Our work suggests that strategies targeting macrophage-mediated monocyte recruitment may represent a promising approach for limiting the chronic inflammation that characterises IBD.

Human monocytes and macrophages regulate immune tolerance via integrin αvß8-mediated TGFß activation.

Monocytes are crucial immune cells involved in regulation of inflammation either directly or via differentiation into macrophages in tissues. However, many aspects of how their function is controlled in health and disease are not understood. Here we show that human blood monocytes activate high levels of the cytokine TGFß, a pathway that is not evident in mouse monocytes. Human CD14+, but not CD16+, monocytes activate TGFß via expression of the integrin αvß8 and matrix metalloproteinase 14, which dampens their production of TNFα in response to LPS. Additionally, when monocytes differentiate into macrophages, integrin expression and TGFß-activating ability are maintained in anti-inflammatory macrophages but down-regulated in pro-inflammatory macrophages. In the healthy human intestine, integrin αvß8 is highly expressed on mature tissue macrophages, with these cells and their integrin expression being significantly reduced in active inflammatory bowel disease. Thus, our data suggest that integrin αvß8-mediated TGFß activation plays a key role in regulation of monocyte inflammatory responses and intestinal macrophage homeostasis.

Culture of Human Monocyte-Derived Macrophages.

The study of human macrophages is often hampered by access to tissue and inability of this cell type to survive in vitro following isolation. The culture of human monocyte-derived macrophages (MDMs) represents a tool to study macrophages, with monocytes known to give rise to tissue macrophages influenced by certain environmental cues. Here we describe a method of culturing monocyte-derived macrophages from CD14+ blood monocytes and polarization toward different macrophage phenotypes.

Viral microRNA effects on persistent infection of human lymphoid cells by polyomavirus SV40.

BACKGROUND: Polyomaviruses, including simian virus 40 (SV40), display evidence of lymphotropic properties. This study analyzed the nature of SV40-human lymphocyte interactions in established cell lines and in primary lymphocytes. The effects of viral microRNA and the structure of the viral regulatory region on SV40 persistence were examined. RESULTS: SV40 DNA was maintained in infected B cell and myeloid cell lines during cell growth for at least 28 days. Limiting dilution analysis showed that low amounts of SV40 DNA (~2 copies per cell) were retained over time. Infected B cells remained viable and able to proliferate. Genome copies of the SV40 microRNA-null mutant persisted at higher levels than the DNA of wild-type viruses. Complex viral regulatory regions produced modestly higher DNA levels than simple regulatory regions. Viral large T-antigen protein was detected at low frequency and at low levels in infected B cells. Following infection of primary lymphocytes, SV40 DNA was detected in CD19+ B cells and CD14+ monocytes, but not in CD3+ T cells. Rescue attempts using either lysates of SV40-infected B lymphocytes, coculture of live cells, or infectious center assays all showed that replication-competent SV40 could be recovered on rare occasions. SV40 infections altered the expression of several B cell surface markers, with more pronounced changes following infections with the microRNA-null mutant. CONCLUSION: These findings indicate that SV40 can establish persistent infections in human B lymphocytes. The cells retain low copy numbers of viral DNA; the infections are nonproductive and noncytolytic but can occasionally produce infectious virus. SV40 microRNA negatively regulates the degree of viral effects on B cells. SIGNIFICANCE: Lymphocytes may serve as viral reservoirs and may function to disseminate polyomaviruses to different tissues in a host. To our knowledge, this report is the first extensive analysis of viral microRNA effects on SV40 infection of human lymphocytes.

Characteristics of nanocrystalline thin films of cadmium sulphide deposited at the water-oil interface.

Thin films of nanocrystalline CdS were obtained at the water-toluene interface by reacting cadmium diethyldithiocarbamate in toluene with aq. Na2S. Three parameters unique to the topical deposition scheme: the effect of column heights, stirring and the action of molecular surfactants are systematically investigated. The obtained nanocrystalline aggregates are characterized by scanning- and transmission electron microscopy, X-ray diffraction and profilometric measurements. Conditions for obtaining smooth device quality thin films have been identified during these experiments.

CD141⁺ myeloid dendritic cells are enriched in healthy human liver.

BACKGROUND & AIMS: Extensive populations of liver immune cells detect and respond to homeostatic perturbation caused by damage, infection or malignancy. Dendritic cells (DCs) are central to these activities, governing the balance between tolerance and immunity. Most of our knowledge about human liver DCs is derived from studies on peritumoral tissue. Little is known about the phenotype and function of DCs, in particular the recently described CD141(+) subset, in healthy human liver and how this profile is altered in liver disease. METHODS: During liver transplantation, healthy donor and diseased explant livers were perfused and hepatic mononuclear cells isolated. Dendritic cell subset frequency and phenotype were characterised in liver perfusates by flow cytometry and the function of CD141(+) DCs was evaluated by mixed lymphocyte reactions (MLRs) and measuring cytokine secretion. RESULTS: Almost one third of liver CD11c(+) myeloid DCs (mDCs) expressed CD141 compared to <5% of circulating mDCs. Hepatic CD141(+) DCs demonstrated pro-inflammatory function in allogeneic MLRs, inducing T cell production of interferon gamma (IFN-γ) and interleukin (IL)-17. While CD123(+) plasmacytoid DCs (pDCs) and CD1c(+) mDCs were expanded in diseased liver perfusates, CD141(+) DCs were significantly depleted. Despite their depletion, CD141(+) DCs from explant livers produced markedly increased poly(I:C)-induced IFN lambda (IFN-λ) compared with donor DCs. CONCLUSIONS: Accumulation of CD141(+) DCs in healthy liver, which are significantly depleted in liver disease, suggests differential involvement of mDC subsets in liver immunity.

Occupational noise exposure of nightclub bar employees in Ireland.

Due to the transposition of the EU Directive 2003/10/EC into Irish Law, the entertainment sector was obligated to comply with the requirements of the Safety, Health and Welfare at Work (General Application) Regulations 2007, Chapter 1 Part 5: Control of Noise at Work since February 2008. Compliance with the Noise Regulations was examined in 9 nightclubs in Ireland. The typical daily noise exposure of 19 bar employees was measured using 2 logging dosimeters and a Type 1 fixed position sound level meter. Physical site inspections identified nightclub noise control measures. Interviews and questionnaires were used to assess the managers and employees awareness of the noise legislation. The average bar employee daily noise exposure (L(EX, 8h)) was 92 dBA, almost 4 times more than the accepted legal limit. None of the venues examined were fully compliant with the requirements of the 2007 Noise Regulations, and awareness of this legislation was limited.