CDC Grand Rounds: Promoting Well-Being and Independence in Older Adults.

Healthy aging is not merely the absence of disease or disability, but requires physical and mental health and ongoing social engagement (1). As the average U.S. life expectancy increases, recognition that public health can play a vital role in promoting healthy, successful aging even in the face of increased prevalence of chronic diseases, including types of dementia, among older adults (i.e., aged ≥65 years) has grown. Furthermore, actively engaging adults in prevention and wellness along with involving their caregivers (i.e., the family and friends of older adults who provide them with unpaid and informal support and services) can serve to prevent or delay the onset of physical disabilities and cognitive decline. Adults often are reluctant to discuss their concerns about worsening memory with their health care providers although such discussions can lead to earlier diagnosis and better care, planning, and support. As advances in public health and health care have helped increase life expectancy, public health professionals and health care providers have the opportunity to improve the quality of life for older adults and their caregivers and reduce the burdens associated with aging.

Identification and analysis of murine pancreatic islet enhancers.

AIMS/HYPOTHESIS: The paucity of information on the epigenetic barriers that are blocking reprogramming protocols, and on what makes a beta cell unique, has hampered efforts to develop novel beta cell sources. Here, we aimed to identify enhancers in pancreatic islets, to understand their developmental ontologies, and to identify enhancers unique to islets to increase our understanding of islet-specific gene expression. METHODS: We combined H3K4me1-based nucleosome predictions with pancreatic and duodenal homeobox 1 (PDX1), neurogenic differentiation 1 (NEUROD1), v-Maf musculoaponeurotic fibrosarcoma oncogene family, protein A (MAFA) and forkhead box A2 (FOXA2) occupancy data to identify enhancers in mouse islets. RESULTS: We identified 22,223 putative enhancer loci in in vivo mouse islets. Our validation experiments suggest that nearly half of these loci are active in regulating islet gene expression, with the remaining regions probably poised for activity. We showed that these loci have at least nine developmental ontologies, and that islet enhancers predominately acquire H3K4me1 during differentiation. We next discriminated 1,799 enhancers unique to islets and showed that these islet-specific enhancers have reduced association with annotated genes, and identified a subset that are instead associated with novel islet-specific long non-coding RNAs (lncRNAs). CONCLUSIONS/INTERPRETATIONS: Our results indicate that genes with islet-specific expression and function tend to have enhancers devoid of histone methylation marks or, less often, that are bivalent or repressed, in embryonic stem cells and liver. Further, we identify a subset of enhancers unique to islets that are associated with novel islet-specific genes and lncRNAs. We anticipate that these data will facilitate the development of novel sources of functional beta cell mass.

The Type 1 Diabetes PhysioLab Platform: a validated physiologically based mathematical model of pathogenesis in the non-obese diabetic mouse.

Type 1 diabetes is an autoimmune disease whose clinical onset signifies a lifelong requirement for insulin therapy and increased risk of medical complications. To increase the efficiency and confidence with which drug candidates advance to human type 1 diabetes clinical trials, we have generated and validated a mathematical model of type 1 diabetes pathophysiology in a well-characterized animal model of spontaneous type 1 diabetes, the non-obese diabetic (NOD) mouse. The model is based on an extensive survey of the public literature and input from an independent scientific advisory board. It reproduces key disease features including activation and expansion of autoreactive lymphocytes in the pancreatic lymph nodes (PLNs), islet infiltration and beta cell loss leading to hyperglycaemia. The model uses ordinary differential and algebraic equations to represent the pancreas and PLN as well as dynamic interactions of multiple cell types (e.g. dendritic cells, macrophages, CD4+ T lymphocytes, CD8+ T lymphocytes, regulatory T cells, beta cells). The simulated features of untreated pathogenesis and disease outcomes for multiple interventions compare favourably with published experimental data. Thus, a mathematical model reproducing type 1 diabetes pathophysiology in the NOD mouse, validated based on accurate reproduction of results from multiple published interventions, is available for in silico hypothesis testing. Predictive biosimulation research evaluating therapeutic strategies and underlying biological mechanisms is intended to deprioritize hypotheses that impact disease outcome weakly and focus experimental research on hypotheses likely to provide insight into the disease and its treatment.

Psychophysical functions for perceived and remembered size.

Separate groups of people estimated the sizes of perceived or of remembered objects. In three independent experiments, both sets of data were well fit by power functions, and the exponent was reliably smaller by remembered than for perceived size.

An analysis of bibliometric data exploring factors affecting research capacity amongst HCPC registered radiographers in England.

INTRODUCTION: Pre-existing studies argue, radiographers are "apathetic" when it comes to research. However, labelling everyone as such seems a gross simplification of a multifaceted issue; especially as current evidence tends to err towards the anecdotal, subjective or is compared to similar professions. Considering this, the SCoR periodically issues a Research Strategy, recognising the necessity to embed/improve research-capacity across all levels of the profession. The aim of this study is to ascertain factors characteristically influencing research-capacity amongst HCPC-registered radiographers in England. METHODS: A purposeful sample of 5 years' bibliometric data from the journal 'Radiography.' A critical and thematic analysis followed based on current peer reviewed journals and grey literature. RESULTS: Of 374 eligible articles and 143 research-active authors (published 2 + articles), collaborations prominently featured (74.9%) across 19 international partnerships. HCPC registrants in England were principal investigator(s) in 49.20% of cases and registrants affiliated with the journal/publisher generally published more than non-affiliates. Preferred topic-areas included 'Education & Research' and 'Technical Practice.' Males published more than females (M = 5.13/F = 3.45). Average length of practice equalled 22.93 years. Outputs mostly originated from HEIs (62.07%), but contributions varied (mean = 10.05/std.deviation = ±17.09); modestly correlating high-REF scoring HEIs (r = 0.330); however, regional workforce ratio(s) proved the strongest indicator (r = 0.601). CONCLUSION: No "one-size-fits all" approach to research-capacity applies; multiple variables affect capacity/activeness. Many seem contingent on extrinsic factors e.g. regional locale, organisational type and culture/support. Personal/professional influences included career status, length of qualification and gender. Future strategies may benefit from refinement; mindful of the dynamics influencing the heterogeneity of the current workforce. Recommendations are that future strategies/studies may benefit from more specific targeting.

Top research priorities in healthcare-associated infection in the UK.

BACKGROUND: There is a mismatch between research questions which are considered to be important by patients, carers and healthcare professionals and the research performed in many fields of medicine. No relevant studies which have assessed research priorities in healthcare-associated infection (HCAI) that have involved patients' and carers' opinions were identified in the literature. AIM: The Healthcare-Associated Infections Priority Setting Partnership was established to identify the top research priorities in the prevention, diagnosis and treatment of HCAI in the UK, considering the opinions of all these groups. METHODS: The methods broadly followed the principles of the James Lind Alliance (JLA) priority setting activity. FINDINGS: In total, 259 unique valid research questions were identified from 221 valid responses to a consultation of patients, carers and healthcare professionals after seeking their opinions for research priorities. The steering committee of the priority setting partnership rationalized these to 50 unique questions. A literature review established that for these questions there were no recent high-quality systematic reviews, high-quality systematic reviews which concluded that further studies were necessary, or the steering committee considered that further research was required despite the conclusions of recent systematic reviews. An interim survey ranked the 50 questions, and the 10 main research priorities were identified from the top 32 questions by consensus at a final priority setting workshop of patients, carers and healthcare professionals using group discussions. CONCLUSIONS: A priority setting process using JLA methods and principles involving patients, carers and healthcare professionals was used to identify the top 10 priority areas for research related to HCAI. Basic, translational, clinical and public health research would be required to address these uncertainties.

Research priorities in mesothelioma: A James Lind Alliance Priority Setting Partnership.

BACKGROUND: In the UK, despite the import and use of all forms of asbestos being banned more than 15 years ago, the incidence of mesothelioma continues to rise. Mesothelioma is almost invariably fatal, and more research is required, not only to find more effective treatments, but also to achieve an earlier diagnosis and improve palliative care. Following a debate in the House of Lords in July 2013, a package of measures was agreed, which included a James Lind Alliance Priority Setting Partnership, funded by the National Institute for Health Research. The partnership brought together patients, carers, health professionals and support organisations to agree the top 10 research priorities relating to the diagnosis, treatment and care of patients with mesothelioma. METHODS: Following the established James Lind Alliance priority setting process, mesothelioma patients, current and bereaved carers, and health professionals were surveyed to elicit their concerns regarding diagnosis, treatment and care. Research questions were generated from the survey responses, and following checks that the questions were currently unanswered, an interim prioritisation survey was conducted to identify a shortlist of questions to take to a final consensus meeting. FINDINGS: Four hundred and fifty-three initial surveys were returned, which were refined into 52 unique unanswered research questions. The interim prioritisation survey was completed by 202 responders, and the top 30 questions were taken to a final meeting where mesothelioma patients, carers, and health professionals prioritised all the questions, and reached a consensus on the top 10. INTERPRETATION: The top 10 questions cover a wide portfolio of research (including assessing the value of immunotherapy, individualised chemotherapy, second-line treatment and immediate chemotherapy, monitoring patients with pleural thickening, defining the management of ascites in peritoneal mesothelioma, and optimising follow-up strategy). This list is an invaluable resource, which should be used to inform the prioritisation and funding of future mesothelioma research.

An immunoassay for the quantitation of cell membrane MHC class II antigen.

A method is described for the quantitative assay of cellular MHC class II proteins. The assay has been developed for special use with epithelial cells isolated from the intestine, but has been successfully used with other cell types. It comprises a competitive immunoassay of anti-class II activity in cell lysates, using IgG-coated Sephacryl S300 as the solid phase and is sensitive over the range 1 ng-1 microgram.

Transforming growth factor-beta messenger RNA and protein in murine colitis.

Using a CD4+ T-cell-transplanted SCID mouse model of colitis, we have analyzed TGF-beta transcription and translation in advanced disease. By in situ hybridization, the epithelium of both control and inflamed tissues transcribed TGF-beta1 and TGF-beta3 mRNAs, but both were expressed significantly farther along the crypt axis in disease. Control lamina propria cells transcribed little TGF-beta1 or TGF-beta3 mRNA, but in inflamed tissues many cells expressed mRNA for both isoforms. No TGF-beta2 message was detected in either control or inflamed tissues. Immunohistochemistry for latent and active TGF-beta1 showed that all cells produced perinuclear latent TGF-beta1. The epithelial cell basal latent protein resulted in only low levels of subepithelial active protein, which co-localized with collagen IV and laminin in diseased and control tissue. Infiltrating cells expressed very low levels of active TGF-beta. By ELISA, very low levels (0-69 pg/mg) of soluble total or active TGF-beta were detected in hypotonic tissue lysates. TGF-beta1 and TGF-beta3 are produced by SCID mouse colon and transcription is increased in the colitis caused by transplantation of CD4+ T-cells, but this does not result in high levels of soluble active protein. Low levels of active TGF-beta may be a factor contributing to unresolved inflammation.

Vaginal and rectal infection of cats with feline immunodeficiency virus.

The objective of this study was to examine the potential of vaginal and rectal mucosal routes for feline immunodeficiency virus (FIV) uptake and infection, as a model of mucosal HIV infection, and to determine the fate of virus at these mucosal sites following transmission of infection. SPF cats were exposed to FIV isolates (PET, GL-8, T637), administered as either cell-associated or cell-free inocula, via the rectum or vagina. Establishment of infection was confirmed by isolation of infectious FIV from peripheral blood mononuclear cells (PBMC), and by presence of FIV proviral DNA in PBMC using a nested polymerase chain reaction. Fate of virus in tissue taken at necropsy from cats infected for 6-48 weeks was assessed by localizing FIV core and envelope proteins, p24 and gp41, using a biotin-streptavidin linked immunoperoxidase (IP) technique. Cells susceptible to infection were identified by an in situ hybridization technique for FIV viral DNA and RNA. Cell-free, as well as cell-associated, virus was infectious across intact vaginal and rectal mucosal surfaces. Transmission was most successful using cell-associated inocula, and via the rectal route. Cells infected with FIV were detected by IP staining in the colon of 6/9 rectally challenged cats and 1/5 vaginally challenged cats. Virus was predominantly localized within the epithelium at the base of the colonic crypts associated with lymphoid aggregates (follicle associated epithelium; FAE), and within the lymphoid follicle itself. Occasional infected cells were also noted within the lamina propria. The distribution of FIV DNA positive cells in the colon was similar to that for FIV antigen whilst FIV RNA positive cells were found more extensively, including within the lamina propria and lymphoid follicle. FIV infected cells were not detected within the vagina, or colonic and ileac lymph nodes. Similar patterns of infected cells were seen in all of the positive cats, indicating that colonic tissues remain persistently actively infected with FIV. We conclude that the FIV/cat model of rectal and vaginal mucosal infection should prove useful for characterizing the mechanism by which HIV infects mucosal surfaces and as a challenge system for the design of vaccines effective at preventing HIV infection via rectal and vaginal routes.

Hib vaccine coverage in children attending day care/nursery school in East York.

This survey assessed Haemophilus influenzae b vaccine coverage and factors affecting compliance in children at increased risk. A telephone survey was conducted of a random sample of 75 children, aged 24-60 months, attending licensed day care/nursery schools in East York. 96% of the sample was surveyed; only 51% were vaccinated. Older children were less likely to be immunized and there were more single-parent families among the non-vaccinated. The age, educational level and country of birth of the parents and the number of siblings were not associated with vaccination status. In 20% of the non-vaccinated group, concern about side effects was the main reason for non-compliance. For 91%, a factor of "moderate/very much concern" was the perceived unimportance of preventing Haemophilus influenzae b illness. Education for parents and physicians, special programs aimed at risk groups, and consideration of mandatory immunization against Haemophilus influenzae b in children attending day care/nursery schools are needed.

A comparison of fluorimetric and colorimetric methods for the determination of alpha-mannosidase activity in bovine plasma.

This paper compares both manual colorimetric and fluorimetric methods for the assay of acidic alpha-mannosidase activity in bovine EDTA plasma. The effect of variable blood volumes per 10 ml blood collecting tube has been investigated, and the colorimetric assay conditions adjusted (for example, zinc concentration in pre-incubation buffer) to cope with low blood volumes resulting in high EDTA/plasma ratios. We have shown that with non-haemolysed EDTA plasma samples, the two methods give similar results, but for haemolysed samples, the colorimetric method yields significantly lower results than those obtained fluorimetrically. Using the manual fluorimetric method, samples can be assayed more rapidly and simply than by using the colorimetric method. This provides definite advantages in a laboratory receiving variable numbers of samples, or which cannot afford the high cost of automated equipment.

Healing of patellar fractures in two kittens.

Two kittens aged between four and five months were presented having sustained patellar fractures. In both cases, healing was subsequently documented radiographically; this has not been reported previously in the literature. One kitten had bilateral patellar fractures - the symptomatic right stifle was treated with a pin and tension-band-wire which later failed, at which point partial patellectomy was performed. The fracture of the left patella was minimally displaced and was treated conservatively. A radiograph of the left patella taken eleven months after initial presentation showed complete healing of the fracture. The second case was treated surgically with a circumferential wire; healing of the fracture was demonstrated radiographically at twelve weeks postoperatively. Radiographic images taken five weeks postoperatively had shown some narrowing of the fracture gap. These two cases demonstrate that bony union of patellar fractures can be documented, given a long enough duration of radiographic follow-up; circumferential wire was an effective treatment in a displaced fracture, and conservative treatment resulted in complete healing of a minimally displaced fracture.

Proinflammatory cytokine synthesis by mucosal fibroblasts from mouse colitis is enhanced by interferon-gamma-mediated up-regulation of CD40 signalling.

Gut mesenchymal fibroblasts form complex phenotypical and functional populations. They participate actively in homeostatic maintenance of the extracellular matrix, epithelial barrier function, repair mechanisms and leucocyte migration. In inflammation, they become activated, change matrix expression and synthesize proinflammatory mediators. Subpopulations of mucosal fibroblasts express CD40 and the aim of this study was to define its role in their proinflammatory function. Stable primary fibroblast lines derived from normal mouse colon and inflamed colon from CD4(+) CD45RB(high)-transplanted SCID mice were used as models to explore the role of mucosal fibroblast CD40 in the inflammatory process. Phenotype correlated with in situ fibroblast phenotype in the tissues of origin. Lines from both sources co-expressed CD40 and Thy1.2 independently of alpha-smooth muscle actin. A subpopulation of CD40(+) fibroblasts from normal colon expressed CD40 at high levels and expression was enhanced by interferon (IFN)-gamma treatment, whereas all CD40(+) fibroblasts from colitis expressed at low levels and expression was unaffected by IFN-gamma treatment. Despite lower-level expression of CD40 by cells from colitis, they secreted constitutively interleukin (IL)-6 and C-C chemokine (CCL)2. Ligation of CD40 enhanced secretion of these mediators and induced secretion of CCL3. CD40 in cells from colitis was more responsive to ligation than CD40 on cells from normal tissue and this sensitivity was amplified selectively by the action of IFN-gamma. We conclude that the inflammatory milieu in colitis induces long-lasting changes in phenotype and proinflammatory function in colonic fibroblasts. In particular, proinflammatory signalling from fibroblast CD40 is amplified synergistically by the Th1 effector T cell cytokine, IFN-gamma.

Systemic administration of the chemokine macrophage inflammatory protein 1alpha exacerbates inflammatory bowel disease in a mouse model.

INTRODUCTION: Exacerbations of inflammatory bowel disease are thought to be related to concurrent infections. As infections are associated with elevated local and serum concentrations of chemokines, we have determined whether systemic administration of the CC chemokine macrophage inflammatory protein 1alpha (MIP-1alpha) exacerbates colitis in a mouse model. METHODS: Colitis was induced in Balb/c mice using trinitrobenzene sulfonic acid (TNBS). Starting four days later, animals received daily intraperitoneal injections of recombinant MIP-1alpha. On day 7, mice were killed and pieces of colon taken for immunohistology and polymerase chain reaction analysis. The direct effects of MIP-1alpha on mucosal T cells and fibroblasts in vitro were also investigated. RESULTS: Systemic administration of MIP-1alpha markedly enhanced colitis with mice developing large transmural ulcers filled with granulation tissue. Treatment resulted in increased numbers of CD4 cells infiltrating the colonic lamina propria, increased interferon gamma (IFN-gamma) levels, and increased transcripts for tumour necrosis factor alpha (TNF-alpha) and matrix metalloproteinase 3 (MMP3). Isolated lamina propria lymphocytes from mice with TNBS colitis contained increased numbers of IFN-gamma and TNF-alpha transcripts when stimulated with MIP-1alpha in vitro. Colonic lamina propria fibroblasts also responded to MIP-1alpha with increased proliferation and decreased collagen 1 synthesis but fibroblast proliferation was not seen in vivo. CONCLUSIONS: These experiments show that increasing serum concentrations of a chemokine, MIP-1alpha, exacerbates immune mediated colitis. The effect seems to be due to the ability of MIP-1alpha to boost Th1 responses in the gut wall. Our findings also suggest a potential pathway by which peripheral infections can exacerbate inflammatory bowel disease.

School performance of children who have experienced maltreatment.

Research has shown that children who have experienced maltreatment are at increased risk of negative developmental outcomes, including difficulties with development of secure attachment relationships and effective peer relationships. However, until recently, few studies have examined the effects of maltreatment on the school performance of children in elementary school through high school. This review highlights published studies that shed light on this question. The need for further research and intervention on the part of occupational therapies who work in school systems is discussed.

Differential control of major histocompatibility complex class II I-Ek alpha protein expression in the epithelium and in subsets of lamina propria antigen-presenting cells of the gut.

In the gut, both the villus epithelium and cells of macrophages and dendritic cell lineages of the lamina propria and Peyer's patches express major histocompatibility complex (MHC) class II glycoproteins and have the potential to present soluble protein antigen. Using mice transgenic for the X and Y promoter deletion mutants of the gene encoding the I-Ek alpha class II protein we have shown: that an intact promoter is essential for expression of I-Ek alpha on the epithelium and lamina propria macrophages; that only the Y box is essential for expression by lamina propria dendritic cells; and that dendritic cells in Peyer's patches are phenotypically more restricted than in the lamina propria and express I-Ek alpha under different regulatory control mechanisms. The results show that different inductive mechanisms exist for class II in distinct mucosal cell populations and provide a model for the analysis of differential antigen handling in the gut mucosa.

Antigen processing by isolated rat intestinal villus enterocytes.

The processing of ovalbumin by isolated rat enterocytes and by splenic adherent cells (SAC) was compared. Paraformaldehyde fixation blocked presentation of ovalbumin to T cells by both cell types when given before, but not after, an antigen pulse. Presentation of ovalbumin by both cell types was blocked by treatment with chloroquine, ammonium chloride or monensin before the antigen pulse. Leupeptin treatment before the antigen pulse inhibited presentation by SAC, but not by enterocytes. Ovalbumin processed by enterocytes was not presented by fixed SAC. Radiolabelled ovalbumin was degraded to small molecular weight fragments by SAC, but not by enterocytes. It is suggested that, compared to the degradative processing seen in macrophages. B cells and dendritic cells, processing of ovalbumin by enterocytes comprises a more subtle, non-degradative mechanism.

Induction of MHC class II gene products in rat intestinal epithelium during graft-versus-host disease and effects on the immune function of the epithelium.

The induction of major histocompatibility complex (MHC) class II expression in the epithelium of the small intestine of the rat during graft-versus-host disease (GVHD) and the effect of this process on the capacity of isolated epithelial cells to present antigen has been investigated. By immunohistology, increased class II (I-A) was noted in lamina propria cells and villus epithelium and de novo expression in crypt epithelium by Day 7 after transfer of parental spleen cells into irradiated hybrid rats. This increased expression reached a maximum by Day 9 or 10. The kinetics of induction of I-E products paralleled those of I-A in villus epithelium, but I-E was not seen in crypt epithelium. Direct radioimmunoassay of class II in isolated villus and crypt epithelial cells revealed a minor peak of class II, particularly in villus cells, very soon after cell transfer which waned and then increased to a second peak at Days 7-9. Assay of presentation of ovalbumin by isolated enterocytes to primed T cells at the peak of class II induction showed that increased class II expression by villus cells mediated enhanced antigen-presenting activity, whereas increases in crypt cell class II did not enable these cells to present ovalbumin.

Effects of passive limb movement on pulmonary ventilation.

This study was undertaken to determine if the observed increase in ventilation during passive limb movement was a reflex hyperventilation or a response to an increased metabolic need for oxygen. Experiments on human volunteers were designed to test the hypothesis that the rapid increase of ventilation at the onset of exercise was due to stimulation of the joints. Results of these studies showed significant increases in ventilation, oxygen consumption, carbon dioxide production, ventilation/oxygen consumption ratio, and heart rate compared to rest and recovery values. The data lead to the conclusion that the rapid increase of ventilation at the onset of exercise is a true hyperventilation and that stimulation of the joints can be a significant contributor to increased pulmonary ventilation.