Replication of adult pancreatic-beta cells cultured on bovine corneal endothelial cell extracellular matrix

Summary A valuable system has been developed to study replication of adult beta cells. Isolated islets from adult rats were partially dispersed and plated on dishes coated with extracellular matrix from bovine-corneal endothelial cells. Within 24 h islet cells attached to the matrix and formed a monolayer. The proportion of insulin-, glucagon-, and somatostatin-containing cells in the cultures was characteristic of whole islets. Function of beta cells was assessed by measuring glucosestimulated insulin release. Insulin release from 7-day-old cultures increased 19-fold after a 16.7 mM glucose challenge indicating that beta-cell function was normal. Cellular replication in the cultures was assessed using the thymidine analogue 5-bromo-2′-deoxyuridine (BrdU). BrdU incorporation was noted in insulin-, glucagon-, and somatostatin-containing cells and also in non-endocrine cells. Among endocrine cells, the majority of BrdU labeling occurred in beta cells. Beta-cell replication potential was assessed using different concentrations of glucose. The incorporation of BrdU into beta cells was affected in a dose-dependent manner by glucose; over a 10-fold increase of beta-cell BrdU labeling was observed when glucose concentration was raised from 5.5 to 16.7 mM. The system proved advantageous for studying the replication potential of adult beta cells.     

Pyrogenic toxins of Staphylococcus aureus in sudden unexpected nocturnal deaths in adults and older children: factors influencing the control of inflammatory responses to toxic shock syndrome toxins

Sudden unexpected nocturnal deaths (SUND) occur in young immigrant workers, mainly from south-east Asia, who are employed in countries such as Singapore and Saudi Arabia. Pyrogenic toxins of Staphylococcus aureus have been identified in two cases of sudden unexpected death in adults in the UK and it has been suggested that these or other toxins with superantigen properties might induce strong inflammatory responses leading to sudden unexpected nocturnal deaths. The objectives of the present study were (1) to assess the levels of antibodies to pyrogenic staphylococcal toxins in the general population, (2) to assess the levels of IgG to the toxins needed to reduce the production of inflammatory mediators by 50% in a model system, (3) to assess in a model system the effects on inflammatory responses to toxic shock syndrome toxin-1 (TSST) of cortisol levels present at night, during the day and under conditions of physiological stress. Enzyme linked immunosorbent assays were used to assess levels of IgG to TSST, staphylococcal enterotoxin A (SEA) and staphylococcal enterotoxin C (SEC). Human buffy coats were used to examine the effect of IgG to the toxins for neutralising activity and the effect of cortisol on induction of inflammatory mediators. Tumour necrosis factor alpha (TNF-alpha) was detected by a bioassay with L929 cells, interleukin-6 (IL-6) and interleukin-10 (IL-10) were measured by an enzyme linked immunosorbent assay. IL-6 and TNF-alpha levels elicited by the toxins were not reduced by night time levels of cortisol (5-10 microg dl(-1)) levels. Day time levels of cortisol (10-20 microg dl(-1)) significantly inhibited IL-6 production but not TNF-alpha in responses. Stress levels of cortisol (40 80 microg dl(-1)) significantly reduced all three cytokines earlier than the normal day time levels. The majority of the population tested had sufficient antibodies to reduce TNF-alpha and IL-6 responses elicited by TSST and SEC in the model system. In the age range in which most sudden unexpected nocturnal death cases occur (20-39 years), males had significantly lower levels of IgG to TSST compared with females. If these toxins play a role in precipitating the series of events leading to sudden unexpected nocturnal death, the higher levels of IgG to the toxins observed in females might explain partly the much higher prevalence of these deaths among men in this age range. If inflammatory responses play a role in sudden unexpected nocturnal death, the inability of the night time levels of cortisol to control IL-6 and TNF-alpha in the model system might reflect these interactions in vivo. The methods developed for detection of the toxins in tissue samples and the quantitative IgG assays for anti-toxins can be applied to investigation of SUND victims to test the hypothesis that some of these deaths are precipitated by pyrogenic staphylococcal toxins.     

Use of the Population Grouping Methodology of the Canadian Institute for Health Information to predict high-cost health system users in Ontario

BACKGROUND:Prior research has consistently shown that the heaviest users account for a disproportionate share of health care costs. As such, predicting high-cost users may be a precondition for cost containment. We evaluated the ability of a new health risk predictive modelling tool, which was developed by the Canadian Institute for Health Information (CIHI), to identify future high-cost cases.METHODS:We ran the CIHI model using administrative health care data for Ontario (fiscal years 2014/15 and 2015/16) to predict the risk, for each individual in the study population, of being a high-cost user 1 year in the future. We also estimated actual costs for the prediction period. We evaluated model performance for selected percentiles of cost based on the discrimination and calibration of the model.RESULTS:A total of 11 684 427 individuals were included in the analysis. Overall, 10% of this population had annual costs exceeding $3050 per person in fiscal year 2016/17, accounting for 71.6% of total expenditures; 5% had costs above $6374 (58.2% of total expenditures); and 1% exceeded $22 995 (30.5% of total expenditures). Model performance increased with higher cost thresholds. The c-statistic was 0.78 (reasonable), 0.81 (strong) and 0.86 (very strong) at the 10%, 5% and 1% cost thresholds, respectively.INTERPRETATION:The CIHI Population Grouping Methodology was designed to predict the average user of health care services, yet performed adequately for predicting high-cost users. Although we recommend the development of a purpose-designed tool to improve model performance, the existing CIHI Population Grouping Methodology may be used — as is or in concert with additional information — for many applications requiring prediction of future high-cost users.

A substantial literature across health systems shows that the highest users of services account for disproportionate shares of the public costs of health care. It has recently been reported that more than three-quarters of individual health care costs in Ontario were incurred by just 10% of the population.¹ Similarly, an Ontario Ministry of Health and Long-Term Care (MOHLTC) analysis of inpatient and home care costs found that the top 5% of patients were responsible for 61% of spending in those domains.² Consistent findings have been reported for Manitoba, Alberta and British Columbia.^3–7Some of the highest-cost cases may be explained by rare, unpredictable events, but others arise in the presence of multiple chronic conditions. Research from the United States has suggested that spending on chronic conditions accounts for the majority of health care expenditures.⁸ Predicting high-cost users may help us to understand and better manage public spending on health care.Cognizant of this need, the Ontario MOHLTC developed a predictive model for high-cost users based on sociodemographic, utilization and clinical diagnostic characteristics.⁹ Although the model performed well, it relied on a coarse categorization of 20 diagnostic variables consisting of broadly defined chapters of the International Statistical Classification of Diseases and Related Health Problems, 10th Revision (ICD-10) and a small number of chronic conditions, which limited its utility for explaining predictions. Moreover, this model is not available for use outside the MOHLTC. As such, there is a need for a predictive model that can be applied more widely by researchers and other stakeholders with an interest in health policy and spending in Canada.The Canadian Institute for Health Information (CIHI) has recently released a new population-based case mix product, the Population Grouping Methodology, which uses diagnoses obtained from patient health care encounters in multiple settings to summarize the universe of diagnosis codes into a clinically meaningful set of 226 health conditions. The grouping and modelling methodologies are described in more detail in Appendix 1 (available at www.cmaj.ca/lookup/suppl/doi:10.1503/cmaj.191297/-/DC1) and in previous reports.^10,11 The CIHI grouping methodology was not designed to predict high-cost cases, and previous work has already shown that the model performs better for low- and moderate-cost users than for highest-cost users (i.e., those with annual costs exceeding $25 000).¹¹ We evaluated the suitability of CIHI’s model for predicting future high-cost users in Ontario by examining the predicted costs for individuals who exceeded the top 10%, 5%, and 1% thresholds of actual cost.     

Blood group, secretor status and susceptibility to bacterial meningitis

Epidemiological evidence is summarized for associations of ABO blood group and secretor status with susceptibility to invasive disease due to capsulate organisms responsible for the majority of bacterial meningitis. Host-parasite interactions that might underly these findings are proposed and evidence to support or refute them provided.