Understanding ageism towards older nursing staff and service users: A systematic mapping review from the perspective of clinical leaders and healthcare managers.

INTRODUCTION: Ageist attitudes negatively affect the quality of care for service users and the working conditions of older nursing staff. Clinical leaders' perceptions of older service users and nursing staff are unknown. AIM: To map research evidence on ageist attitudes in healthcare towards service users and older nursing staff, from the leadership perspective. MATERIAL AND METHODS: A systematic mapping review with database searches in March 2021 and May 2023). Nineteen articles (qualitative n = 13, quantitative n = 3, mixed methods n = 3) were analysed and mapped to the Nurse Executive Capability Framework. RESULTS: Future planning, team building, and self-awareness are leadership categories requiring consideration. Guidelines and policies addressing the holistic needs of older service users and older nursing staff are lacking. CONCLUSIONS: Organizational planning, team building, and self-awareness are crucial for the holistic care of older service users and for creating attractive workplaces for older nursing staff.

Factors Associated with Healing Outcomes in Primary Care Patients with Diabetic Foot Ulcers: A Retrospective Study in a Multiethnic Sample.

OBJECTIVE: To identify and determine patient- and ulcer-related factors associated with healing outcomes within 3 months for patients with diabetic foot ulcer (DFU) in a multiethnic primary care sample. METHODS: Retrospective data were collected over 3 months from 520 primary care patients with a DFU between April 1, 2016 and March 31, 2017. Multivariable prevalence ratios (PRs) were calculated using Poisson regression to find associations between patient- and ulcer-related factors and healing outcomes. RESULTS: Most patients were male (66%) and Chinese (49.8%) and had a diabetes mellitus duration longer than 5 years (81.8%). Toe ulcers (64%) were most common. Healing occurred for 33.9% of participants; 19.1% and 1.5% underwent minor and major amputation, respectively. Wound sizes between 1 and 10 cm2 (PR, 0.61; 95% confidence interval [CI], 0.46-0.76; P < .001) and over 10 cm2 (PR, 0.55; 95% CI, 0.33-0.76; P = .003), ulcer duration 6 months or longer (PR, 0.36; 95% CI, 0.19-0.53; P < .001), ischemic ulcers (PR, 0.54; 95% CI, 0.22-0.86; P = .044), and neuroischemic ulcers (PR, 0.73; 95% CI, 0.53-0.93; P = .027) were negatively associated with healing outcomes. Women were more likely to experience healing (PR, 1.18; 95% CI, 0.91-1.45; P = .157). CONCLUSIONS: Ulcer healing varied by sex and was affected by wound size, wound duration, and ischemic etiology, regardless of ethnicity. Prompt attention to these risk factors may reduce healing time. Further studies are warranted to elucidate the mechanism underlying sex differences in association with DFU healing.

Health-related quality of life and chronic wound characteristics among patients with chronic wounds treated in primary care: A cross-sectional study in Singapore.

Chronic wounds commonly decrease patients' quality of life. Understanding how chronic wounds impact a patient's health-related quality of life (HRQoL) is important for healthcare service delivery and treatment management. This study explored HRQoL among patients suffering from chronic wounds and investigated associations with patients' socio-demographics and wound characteristics. Two hundred and thirty-three patients across six primary care clinics were assessed and responded to a survey that collected information on socio-demographic, wound characteristics, and HRQoL using the EQ-5D-5L instrument. Data were analysed by descriptive statistics and generalised linear models. The mean age of patients was 61.2 (SD: 14.6) years; 68.2% were males; and 61.8% were of Chinese origin. Arterial ulcers had the greatest negative impact on HRQoL related to mobility, self-care, pain/discomfort and anxiety/depression, and the lowest VAS mean score 62.31 (SD: 28.3; range: 0-100) indicating the worst health. HRQoL related to mobility was significantly associated with age (ß = 0.008, P < .001), non-Chinese ethnicity (ß = 0.25, P = .001), mixed ulcers (ß = -0.41, P = .022), atypical hard-to-heal wounds (ß = -0.38, P = .021), wounds with low (ß = 0.24, P = .044) to moderate (ß = 0.29, P = .018) exudate level, and a wound duration ≥6 months (ß = 0.19, P = .033). The findings can be used to improve healthcare delivery for patients with chronic wound to optimise their HRQoL.

Concordance Between Physician-rated and Caregiver-perceived Disease Severity in Children with Atopic Dermatitis: A Cross-sectional Study.

This study examined concordance between caregiver-reported and physician-rated estimates of severity of atopic dermatitis (AD) in paediatric patients and explored potential explanatory factors. Physician-reported severity of AD was retrieved from medical records, while caregiver-reported disease severity and sociodemographic data were obtained through a survey that also collected information on out-of-pocket expenses due to AD. There was 38.5% (95% confidence interval (95% CI) 30.1, 43.5) disagreement between physician and caregivers with regards to both underestimating and overestimating the condition. A duration since AD diagnosis shorter than 6 months showed higher concordance (kappa: 44.4%; 95% CI 30.6, 58.2) between caregiver and physician estimates of AD severity compared with a duration of 6 months or longer. Caregivers underestimating their child's AD accounted for 27.7% among all participants, while 10.8% overestimated the severity of AD compared with physicians. Factors significantly associated with caregiver's underestimation of disease severity were age of the child and time since disease diagnosis. Comparison of concordance between caregiver-reported and physician-rated estimates of severity of AD in paediatric patients revealed a tendency amongst caregivers to underestimate severity of AD. This information may have clinical implications for treatment outcomes if caregivers fail to adhere to medical advice.

Associations between health-related quality of life and health care costs among children with atopic dermatitis and their caregivers: A cross-sectional study.

BACKGROUND/OBJECTIVES: Atopic dermatitis is associated with a decreased health-related quality of life and contributes to substantial health care costs. It is important to understand what accelerates health care costs to inform various stakeholders, so they can effectively meet health care needs. This cross-sectional study aims to explore associations between health-related quality of life, health care costs, and cost-accelerating variables. METHODS: Information on health-related quality of life (HRQoL) was collected through generic and disease-specific instruments from caregivers and children <16 years of age with a physician-confirmed diagnosis of atopic dermatitis. The economic impact of atopic dermatitis was evaluated by analyzing information on health service utilization and other health care costs related to managing the condition. RESULTS: Children with high impact on health-related quality of life presented an annual health care cost of US$ 3787 compared with US$ 2548 for moderately impacted and US$ 2258 among children for which the condition had low impact. The severity of atopic dermatitis, disease duration, and a lower health-related quality of life was associated with greater health care costs. Analyses of subdomains of health-related quality of life revealed correlations between "mood" and "personal relationships" on one hand and caregivers' physical health and health care costs on the other hand. CONCLUSIONS: Highly affected HRQoL is associated with increased health care costs and could be a valuable complement to traditional severity assessments. By using HRQoL instruments, burdens and symptoms beyond severity can be identified and addressed with interventions to increase HRQoL and subsequently reduce health care costs.

The humanistic and economic burden of chronic wounds: A systematic review.

Chronic wounds are a health problem that have devastating consequences for patients and contribute major costs to healthcare systems and societies. To understand the magnitude of this health issue, a systematic review was undertaken. Searches were conducted in MEDLINE, EMBASE, EBM Reviews and Cochrane library, CINAHL, EBSCO, PsycINFO, and Global Health databases for articles published between 2000 and 2015. Included publications had to target adults (≥18 years of age), state wound chronicity (≥3 weeks) and/or label the wounds as chronic, complex, hard-to-heal, or having led to an amputation. The review excluded studies that did not present data on generic health-related quality of life and/or cost data, case studies, randomized controlled trials, economic modeling studies, abstracts, and editorials. Extracted data were summarized into a narrative synthesis, and for a few articles using the same health-related quality of life instrument, average estimates with 95% confidence intervals were calculated. Thirty articles met the inclusion criteria. Findings revealed that health-related quality of life was lowest for physical pathologies, and based on average estimates were scores most inferior in the domain physical role for both patients with chronic wounds and for those with wound-related amputations. The cost burden was mainly attributed to amputations for patients also comorbid with diabetes, where the cost for hospitalization ranged from US$12,851 to US$16,267 (median) for this patient group. Patients with chronic wounds have poor health-related quality of life in general and wound-related costs are substantial. Development and implementation of wound management strategies that focus on increasing health-related quality of life and effectively reduce costs for this patient group are urgently needed.

CDKN2B expression and subcutaneous adipose tissue expandability: possible influence of the 9p21 atherosclerosis locus.

Risk alleles within a gene desert at the 9p21 locus constitute the most prevalent genetic determinant of cardiovascular disease. Previous research has demonstrated that 9p21 risk variants influence gene expression in vascular tissues, yet the biological mechanisms by which this would mediate atherosclerosis merits further investigation. To investigate possible influences of this locus on other tissues, we explored expression patterns of 9p21-regulated genes in a panel of multiple human tissues and found that the tumor suppressor CDKN2B was highly expressed in subcutaneous adipose tissue (SAT). CDKN2B expression was regulated by obesity status, and this effect was stronger in carriers of 9p21 risk alleles. Covariation between expression of CDKN2B and genes implemented in adipogenesis was consistent with an inhibitory effect of CDKN2B on SAT proliferation. Moreover, studies of postprandial triacylglycerol clearance indicated that CDKN2B is involved in down-regulation of SAT fatty acid trafficking. CDKN2B expression in SAT correlated with indicators of ectopic fat accumulation, including markers of hepatic steatosis. Among genes regulated by 9p21 risk variants, CDKN2B appears to play a significant role in the regulation of SAT expandability, which is a strong determinant of lipotoxicity and therefore might contribute to the development of atherosclerosis.

The TGR5 gene is expressed in human subcutaneous adipose tissue and is associated with obesity, weight loss and resting metabolic rate.

Bile acids have emerged as a new class of signaling molecules that play a role in metabolism. Studies in mice have shown that the bile acid receptor TGR5 mediates several of these effects but the metabolic function of TGR5 in humans is less well established. Here we show that human adipose tissue TGR5 expression is positively correlated to obesity and reduced during diet-induced weight loss. Adipose tissue TGR5 expression was also positively correlated to resting metabolic rate. Our study indicates that human adipose tissue contributes to the TGR5 mediated metabolic effects of bile acids and plays a role in energy expenditure.

Nurses' general attitudes and caregiving-specific perceptions toward the oldest-old: A nationwide survey.

BACKGROUND: Despite the growing population of older adults worldwide, nurses may harbor negative attitudes toward older patients that might risk patient safety. Most studies have investigated general attitudes toward older adults. Only a few have focused on nurses' perceptions of caring for older patients, and there has been little focus on the oldest-old (aged ≥80 years), an age group that is particularly affected by negative attitudes. Emerging models to combat this issue suggest that education about aging and interaction with older adults are key factors. These models have primarily been evaluated using students, not healthcare professionals. OBJECTIVE: To examine if and to what extent education (training and education about working with older people) and interaction (professional and personal contact with older adults) are associated with nurses' general attitudes toward older persons and their perceptions of caring for older patients (aged ≥80 years). DESIGN: A cross-sectional survey study. PARTICIPANTS: A nationwide sample of 2294 Swedish registered nurses with background characteristics reflective of the national nursing population. METHODS: Emails with study information and survey links were distributed via the Swedish Association of Health Professionals' registry. The survey contained one instrument that measures general attitudes toward older adults and one that measures perceptions of caring for older patients. Following the Positive Education about Aging and Contact Experiences model, items about participants' education and their professional and personal interaction with the oldest-old adults were also included. Descriptive and comparative analyses were conducted. Multiple regression models used general attitudes and caregiving-specific perceptions as dependent variables in each model. RESULTS: Nurses' general attitudes and caregiving-specific perceptions were correlated. Regression analyses revealed that the education and interaction variables were independently associated with general attitudes and caregiving-specific perceptions, but these associations followed different directions. An older family member/close friend, extensive work experience, and working primarily in administration were independently associated with more positive general attitudes toward the oldest-old adults. Working in home care or assisted living, having education in geriatrics/gerontology, and frequent interaction with older patients were associated with less favorable general attitudes but more positive perceptions of caring for the oldest-old patients. CONCLUSIONS: Our results highlight the complex relationship between nurses' general attitudes and their caregiving-specific perceptions toward the oldest-old persons. Education in geriatrics/gerontology and interaction with older adults are warranted to improve nurses' caregiving-specific perceptions. Appropriate scales that consider this complexity beyond general attitudes are needed to capture caregiving-specific and contextual perceptions.

Health-Related Quality of Life and Work Satisfaction in Working-Aged Adults Pre- and Post-Cochlear Implant: A Longitudinal Study.

Hearing loss is a growing public health concern associated with decreased health-related quality of life (HRQoL) and a negative impact on work life. Knowledge about the long-term benefits for patients receiving cochlear implants may provide knowledge imperative for policymakers to promote better HRQoL and working life outcomes for individuals with hearing loss. The purpose of this study was to explore how HRQoL, hearing disabilities, and work satisfaction outcomes changed in working-aged adults with severe to profound hearing loss from pre- to post-receiving a cochlear implant (CI) between the baseline, year one, and year two. This longitudinal study used Cochlear's Implant Recipient Observational Study (IROS) registry data to assess HRQoL, hearing disabilities, and work satisfaction in 18-65-year-old CI recipients. Data were collected pre- and post-implantation at baseline, year one, and year two follow-up. One hundred and twenty-seven CI recipients participated in the study. Significant improvements were observed for HRQoL outcomes for hearing, speech, emotion, and health utility post-implant. Overall hearing disability decreased post-CI, and work satisfaction improved. With the increasing prevalence of hearing disabilities, this is pertinent knowledge that supports the use of CIs for hearing rehabilitation which may promote better HRQoL and work satisfaction.

A Scoping Review of End-of-Life Communication in International Palliative Care Guidelines for Acute Care Settings.

CONTEXT: End-of-life communication in acute care settings can be challenging and many patients and families have reported low satisfaction with those conversations. OBJECTIVE: To explore existing guidelines around palliative care to increase current understanding of end-of-life communication processes applicable to the acute care setting. METHODS: A scoping review following the method of Arksey and O'Malley was undertaken to identify eligible documents and thematically summarize findings. Web sites of government authorities, departments, and ministries of health as well as palliative care organizations were searched as were MEDLINE, CINAHL (EBSCOhost), EMBASE, Cochrane Library, Joanna Briggs Institute, and PsycINFO databases. Searches were limited to documents published between January 2009 and August 2019 that were nondisease specific and applicable to the acute care setting. RESULTS: Thirteen guidelines from nine different countries were identified. Thematic analysis produced eight themes: 1) The purpose and process of end-of-life communications, 2) cognitive understanding and language in end-of-life communication, 3) legal aspects of end-of-life communication, 4) conflicts and barriers related to end-of-life care, 5) end-of-life communication related to medical record documentation, 6) healthcare professionals' responsibilities and collaboration, 7) education and training, and 8) policies, guidelines, and tools for end-of-life communications. CONCLUSIONS: Palliative and end-of-life guidelines applicable to acute care settings outline the purpose of end-of-life communication and address how, when, and by whom such conversations are best initiated and facilitated. How guidelines are developed and what aspects of communications are included and emphasized may differ across countries related to role differences of physicians and nurses and national laws and regulations.

Digital health professions education on chronic wound management: A systematic review.

BACKGROUND: Continuing education is crucial for healthcare professionals to keep up with research but attending classroom lectures is a major barrier. Chronic wound management is increasingly relevant for continuous professional training. Digital education offers learning tailored to individual needs and could be an effective alternative to healthcare professionals' training. However, the effectiveness of digital education for chronic wound management training has not been explored. OBJECTIVES: To assess the effectiveness of digital education in improving healthcare professionals' knowledge, attitudes, practical skills and behaviour change on chronic wound management, and their satisfaction with the intervention. DESIGN: This systematic review follows Cochrane methodology and is one of a series of reviews on the use of digital education for health professions education. Protocol registration: PROSPERO CRD42018109971 DATA SOURCES: Searches were conducted in MEDLINE, Embase, Web of Science, ERIC, PsycINFO, CINAHL, CENTRAL, and ProQuest Dissertation and Theses Database. REVIEW METHODS: We included randomised control trials, cluster randomised control trials and quasi-randomised control trials comparing digital or blended education with traditional learning, no intervention or other forms of digital or blended education for pre- or post-registration healthcare professionals in chronic wound management. A narrative summary of findings is presented. RESULTS: Seven studies (1,404 participants) were included. All studies investigated interventions for nursing students or professionals working in hospitals or community settings, and all but one study focused on pressure ulcers. Five studies (935 participants) assessed post-intervention knowledge, and indicated that digital education was more effective than no intervention, while blended learning was superior to exclusive digital education. Three studies (543 participants) assessed post-intervention skills and reported mixed results. One study (140 participants) compared post-intervention behaviour change and satisfaction with blended and online digital education, and reported no difference in behaviour between the groups, and higher satisfaction with blended education. For knowledge retention up to six months, digital education was more effective than no intervention, while blended learning was superior to digital education. The risk of bias in included studies was mostly high or unclear. CONCLUSIONS: Digital education on chronic wound management appears to be less effective than blended education and more effective than no intervention in improving knowledge among nurses and nursing students. Data for other outcomes is scarce and inconclusive. Future studies should assess participants' skills, attitudes, satisfaction and behaviour change; cost-effectiveness and potential untoward effects of digital education, compare digital education to other learning modalities and include other healthcare professionals in diverse clinical settings.

Association of serum amyloid A levels with adipocyte size and serum levels of adipokines: differences between men and women.

The aim of this study was to characterize the association between adipocyte enlargement and circulating levels of serum amyloid A (SAA). Furthermore, we wanted to search for possible associations with measures of glycemic control and levels of circulating adipokines and/or inflammatory markers in men and women with a large range in body mass index. The study cohort consisted of 167 subjects, 114 non-diabetic and 53 with Type 2 diabetes. Adipocyte diameter as well as circulating levels of SAA, C-reactive protein (CRP), adiponectin, leptin, interleukin-6, tumor necrosis factor alpha, glucose and insulin were measured. Women had higher serum levels of SAA than men (p=0.044). SAA levels were weakly but positively correlated with BMI (p=0.043) and % body fat (p=0.027) in all subjects as well as subcutaneous adipocyte diameter (p=0.034) in women. Furthermore, in all subjects we found correlations between SAA levels and levels of CRP (p<0.001), interleukin-6 (p<0.001), leptin (p=0.003), insulin (p=0.006), HbA1c (p=0.02) and HOMA-IR (p=0.002). A majority of the correlations were strongest in women. In conclusion, serum levels of SAA are strongly correlated with serum levels of inflammatory markers as well as measures of glycemic control. There seems to be large sex differences in these associations suggesting that sex-specific factors need to be considered when analyzing SAA levels in relation to metabolic disease.

Prevalence of chronic wounds in the general population: systematic review and meta-analysis of observational studies.

PURPOSE: Chronic wounds are a major public health challenge, but little is known about the true burden with studies reporting different estimates because of disparities in study designs and measurement methods. This hampers efficient resource allocation, planning, and improvement of wound care. METHODS: Our study aimed to pool prevalence estimates from a global perspective by systematically carrying out searches in MEDLINE, EMBASE, Cochrane, CINAHL, Global Health, and PsycINFO databases for articles reporting the prevalence of chronic wounds in adults, from January 2000 to June 2018. The included publications had to define wound chronicity by duration (≥3 weeks), and/or labeling the wounds as chronic, complex, or hard-to-heal. RESULTS: Seventeen studies met the inclusion criteria, and 11 studies analyzing chronic wounds in the general population were included in random effects meta-analyses to calculate pooled prevalence. Chronic wounds of mixed etiologies (n = 3) showed a pooled prevalence of 2.21 per 1000 population, and for chronic leg ulcers (n = 9), the prevalence was estimated at 1.51 per 1000 population. CONCLUSIONS: Our findings, aligned to previous studies reporting point prevalence of chronic wounds identified within the healthcare system, showed that the vast majority of chronic wounds in epidemiological studies are made up by chronic leg ulcers.

Genome-wide association meta-analysis of functional outcome after ischemic stroke.

OBJECTIVE: To discover common genetic variants associated with poststroke outcomes using a genome-wide association (GWA) study. METHODS: The study comprised 6,165 patients with ischemic stroke from 12 studies in Europe, the United States, and Australia included in the GISCOME (Genetics of Ischaemic Stroke Functional Outcome) network. The primary outcome was modified Rankin Scale score after 60 to 190 days, evaluated as 2 dichotomous variables (0-2 vs 3-6 and 0-1 vs 2-6) and subsequently as an ordinal variable. GWA analyses were performed in each study independently and results were meta-analyzed. Analyses were adjusted for age, sex, stroke severity (baseline NIH Stroke Scale score), and ancestry. The significance level was p < 5 × 10-8. RESULTS: We identified one genetic variant associated with functional outcome with genome-wide significance (modified Rankin Scale scores 0-2 vs 3-6, p = 5.3 × 10-9). This intronic variant (rs1842681) in the LOC105372028 gene is a previously reported trans-expression quantitative trait locus for PPP1R21, which encodes a regulatory subunit of protein phosphatase 1. This ubiquitous phosphatase is implicated in brain functions such as brain plasticity. Several variants detected in this study demonstrated suggestive association with outcome (p < 10-5), some of which are within or near genes with experimental evidence of influence on ischemic stroke volume and/or brain recovery (e.g., NTN4, TEK, and PTCH1). CONCLUSIONS: In this large GWA study on functional outcome after ischemic stroke, we report one significant variant and several variants with suggestive association to outcome 3 months after stroke onset with plausible mechanistic links to poststroke recovery. Future replication studies and exploration of potential functional mechanisms for identified genetic variants are warranted.

A Genome-wide Study of Common and Rare Genetic Variants Associated with Circulating Thrombin Activatable Fibrinolysis Inhibitor.

Thrombin-activatable fibrinolysis inhibitor (TAFI) plays a central role in haemostasis, and plasma TAFI concentrations are heritable. Candidate gene studies have identified several variants within the gene encoding TAFI, CPB2, that explain part of the estimated heritability. Here, we describe an exploratory genome-wide association study to identify novel variants within and outside of the CPB2 locus that influence plasma concentrations of intact TAFI and/or the extent of TAFI activation (measured by released TAFI activation peptide, TAFI-AP) amongst 3,260 subjects from Southern Sweden. We also explored the role of rare variants on the HumanExome BeadChip. We confirmed the association with previously reported common variants in CPB2 for both intact TAFI and TAFI-AP, and discovered novel associations with variants in putative CPB2 enhancers. We identified a gene-based association with intact TAFI at CPB2 (PSKAT-O = 2.8 × 10-8), driven by two novel rare nonsynonymous single nucleotide polymorphisms (SNPs; I420N and D177G). Carriers of the rare variant of D177G (rs140446990; MAF 0.2%) had lower intact TAFI and TAFI-AP concentrations compared with non-carriers (intact TAFI, geometric mean 53 vs. 78%, PT-test = 5 × 10-7; TAFI-AP 63 vs. 99%, PT-test = 7.2 × 10-4). For TAFI-AP, we identified a genome-wide significant association at an intergenic region of chromosome 3p14.1 and five gene-based associations (all PSKAT-O < 5 × 10-6). Using well-characterized assays together with a genome-wide association study and a rare-variant approach, we verified CPB2 to be the primary determinant of TAFI concentrations and identified putative secondary loci (candidate variants and genes) associated with intact TAFI and TAFI-AP that require independent validation.

Discrimination and scoring using small sets of genes for two-sample microarray data.

Comparison of gene expression for two groups of individuals form an important subclass of microarray experiments. We study multivariate procedures, in particular use of Hotelling's T2 for discrimination between the groups with a special emphasis on methods based on few genes only. We apply the methods to data from an experiment with a group of atopic dermatitis patients compared with a control group. We also compare our methodology to other recently proposed methods on publicly available datasets. It is found that (i) use of several genes gives a much improved discrimination of the groups as compared to one gene only, (ii) the genes that play the most important role in the multivariate analysis are not necessarily those that rank first in univariate comparisons of the groups, (iii) Linear Discriminant Analysis carried out with sets of 2-5 genes selected according to their Hotelling T2 give results comparable to state-of-the-art methods using many more genes, a feature of our method which might be crucial in clinical applications. Finding groups of genes that together give optimal multivariate discrimination (given the size of the group) can identify crucial pathways and networks of genes responsible for a disease. The computer code that we developed to make computations is available as an R package.

Genome-wide methylation profiling identifies novel methylated genes in neuroblastoma tumors.

Neuroblastoma is a very heterogeneous tumor of childhood. The clinical spectra range from very aggressive metastatic disease to spontaneous regression, even without therapy. Aberrant DNA methylation pattern is a common feature of most cancers. For neuroblastoma, it has been demonstrated both for single genes as well as genome-wide, where a so-called methylator phenotype has been described. Here, we present a study using Illumina 450K methylation arrays on 60 neuroblastoma tumors. We show that aggressive tumors, characterized by International Neuroblastoma Risk Group (INRG) as stage M, are hypermethylated compared to low-grade tumors. On the contrary, INRG stage L tumors display more non-CpG methylation. The genes with the highest number of hypermethylated CpG sites in INRG M tumors are TERT, PCDHGA4, DLX5, and DLX6-AS1. Gene ontology analysis showed a representation of neuronal tumor relevant gene functions among the differentially methylated genes. For validation, we used a set of independent tumors previously analyzed with the Illumina 27K methylation arrays, which confirmed the differentially methylated sites. Top candidate genes with aberrant methylation were analyzed for altered gene expression through the R2 platform ( http://r2.amc.nl), and for correlations between methylation and gene expression in a public dataset. Altered expression in nonsurvivors was found for the genes B3GALT4 and KIAA1949, CLIC5, DLX6-AS, TERT, and PIRT, and strongest correlations were found for TRIM36, KIAA0513, and PIRT. Our data indicate that methylation profiling can be used for patient stratification and informs on epigenetically deregulated genes with the potential of increasing our knowledge about the underlying mechanisms of tumor development.

Adipose tissue-derived human serum amyloid a does not affect atherosclerotic lesion area in hSAA1+/-/ApoE-/- mice.

Chronically elevated serum levels of serum amyloid A (SAA) are linked to increased risk of cardiovascular disease. However, whether SAA is directly involved in atherosclerosis development is still not known. The aim of this study was to investigate the effects of adipose tissue-derived human SAA on atherosclerosis in mice. hSAA1+/- transgenic mice (hSAA1 mice) with a specific expression of human SAA1 in adipose tissue were bred with ApoE-deficient mice. The hSAA1 mice and their wild type (wt) littermates were fed normal chow for 35 weeks. At the end of the experiment, the mice were euthanized and blood, gonadal adipose tissue and aortas were collected. Plasma levels of SAA, cholesterol and triglycerides were measured. Atherosclerotic lesion areas were analyzed in the aortic arch, the thoracic aorta and the abdominal aorta in en face preparations of aorta stained with Sudan IV. The human SAA protein was present in plasma from hSAA1 mice but undetectable in wt mice. Similar plasma levels of cholesterol and triglycerides were observed in hSAA1 mice and their wt controls. There were no differences in atherosclerotic lesion areas in any sections of the aorta in hSAA1 mice compared to wt mice. In conclusion, our data suggest that adipose tissue-derived human SAA does not influence atherosclerosis development in mice.

Endogenous Acute Phase Serum Amyloid A Lacks Pro-Inflammatory Activity, Contrasting the Two Recombinant Variants That Activate Human Neutrophils through Different Receptors.

Most notable among the acute phase proteins is serum amyloid A (SAA), levels of which can increase 1000-fold during infections, aseptic inflammation, and/or trauma. Chronically elevated SAA levels are associated with a wide variety of pathological conditions, including obesity and rheumatic diseases. Using a recombinant hybrid of the two human SAA isoforms (SAA1 and 2) that does not exist in vivo, numerous in vitro studies have given rise to the notion that acute phase SAA is a pro-inflammatory molecule with cytokine-like properties. It is however unclear whether endogenous acute phase SAA per se mediates pro-inflammatory effects. We tested this in samples from patients with inflammatory arthritis and in a transgenic mouse model that expresses human SAA1. Endogenous human SAA did not drive production of pro-inflammatory IL-8/KC in either of these settings. Human neutrophils derived from arthritis patients displayed no signs of activation, despite being exposed to severely elevated SAA levels in circulation, and SAA-rich sera also failed to activate cells in vitro. In contrast, two recombinant SAA variants (the hybrid SAA and SAA1) both activated human neutrophils, inducing L-selectin shedding, production of reactive oxygen species, and production of IL-8. The hybrid SAA was approximately 100-fold more potent than recombinant SAA1. Recombinant hybrid SAA and SAA1 activated neutrophils through different receptors, with recombinant SAA1 being a ligand for formyl peptide receptor 2 (FPR2). We conclude that even though recombinant SAAs can be valuable tools for studying neutrophil activation, they do not reflect the nature of the endogenous protein.