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Foodborne diseases constitute a major concern in societies, and their causes are aimed to be identified and minimized. Only in the last few years, this is encouraged by the application of risk assessment, management, and communication. This work presents a probabilistic quantitative microbiological risk assessment and management of Listeria monocytogenes in ready-to-eat lettuce salads in Spain. For risk assessment, a guideline provided by Codex Alimentarius was followed. Food chain was modeled from processing of raw material at the factory up to consumption. Different assumptions were made to describe the variables of the model by probability distributions or mathematical models. Monte Carlo simulations of the model were run to estimate the number of cases in low-risk and high-risk populations. Although results deviated from the number of cases observed in Spain, given an ideal situation of 100% compliance of the microbiological criterion ≤100 cfu/g throughout the shelf-life of the product, the resulting number of cases was near the real situation. From the 4 risk management measures simulated, the injection of a mixture of gases into packages at manufacture (CO2 about 5.5%, O2 about 3%, and N2 for the balance) was the most effective in reducing the number of cases, followed by 4 d of storage at home and prevention of high-risk consumers from consumption of ready-to-eat lettuce salads. More research and cooperation between different stakeholder organizations are needed in order to progressively improve the model. With this work, a breakthrough has been made with regards to risk assessment and management procedures and implementation.

Legislation on food safety has led towards the standardization of food productions which, together with the existing quality certifications, aim to increase the level of protection of public health. It is recognized the need for the agri-food industry to have tools to harmonize their productions and to adequately manage their quality systems in order to improve consumers’ confidence. The implementation of microbiological criteria is focused on facilitating this harmonization by enabling the discrimination of defective lots and acting as control tools at industrial level. Therefore, knowledge of the principles, components and factors influencing the efficiency of microbiological criteria may be helpful to better understand the consequences of their application. In the present study the main principles, methodologies and applications of microbiological criteria in foods are addressed for their implementation as a part of the management quality systems of agrifood industries. In addition, potential limitations and impact of microbiological criteria on food safety are discussed. Finally, an assessment of the performance of microbiological criteria at EU level in berries is described for the compliance of the socalled risk-based metrics, namely Performance Objectives and Food Safety Objectives.

Strawberry production represents an agriculture sector of high relevance for the Spanish economy, due to their strawberries, highly appreciated for their organoleptic characteristics and health benefits. However, the reported outbreaks in different countries by enteric pathogens associated with this product have increased the safety concerns among different stakeholders. A number of factors and pathways for introducing pathogens in the strawberry production chain has been investigated. However, information on the potential internalization of enteric pathogens in strawberries is still scarce. The present study aimed to evaluate the potential internalization of Salmonella enterica sv Thompson via root in strawberry plants under the application of successive contamination events through capillary irrigation. Strawberry plants were organized in three groups (16 plants per group), and they were submitted to different contamination scenarios; groups 1, 2 and 3 were irrigated 1, 2 and 3 times respectively, with 100 mL of water containing Salmonella Thompson (8.8 log10 CFU/mL) every other day for 9 days. Fruits, calyces, and leaves were analyzed during the experiment, while roots were analyzed on the 9th day. The results showed that all calyces analyzed were negative, while one strawberry sample (group 3; day 8), four leaves samples (groups 2 and 3; day 8) and one root sample (group 3; day 9) were positive for Salmonella Thompson, representing the 0.85% (95% C.I.: [0.02–4.63%]), 8.88% (95% C.I.: [2.48–21.22%]), and 2.22% (95% C.I.: [0.06–11.77%]), respectively, of the overall samples analyzed. A significant association between the increased frequency of contamination (three irrigations) and the presence of the pathogen in the samples was elucidated. Our findings, in comparison with other studies, also point out the importance of watering by drip irrigation as a more hygienic and safer agronomic practice than systems like sprinkling irrigation.

A tool able to quantitatively assess the fate of potential pathogenic microorganisms in foods along the food chain and their impact on public health is highly valuable for food safety decision-makers. The aim of this work was to present an overview of the Predictive Microbiology software MicroHibro, which is able to assess the evolution of potential pathogens and spoilage microorganisms along the food chain, providing estimates for the exposure level and risk associated with a food product. The application is built on an extensive Predictive Microbiology Model Data Base (PMDB) including kinetic processes like growth, inactivation, transfer as well as dose-response models. PMDB can be populated with new models by using an on-line tool in combination with a standardized method for describing Predictive Microbiology models. This enables MicroHibro to be easily updated, increasing its applicability and use. Estimation of microbial risk associated with a food product can be achieved, in MicroHibro, by describing steps in any food chain using four different microbial processes (growth, inactivation, transfer and partitioning). As a result, an estimate of the concentration and prevalence of microorganisms in the food of interest as well as attendant risk are provided. Also, MicroHibro allows comparing different predictive models and validate them by introducing user's data. In this paper, examples are provided to illustrate how predictive models can be incorporated in MicroHibro, and then, used to develop a Quantitative Microbial Risk Assessment model. The use of expert computational systems is a powerful tool for supporting food safety and quality activities by Health Authorities and the food industry. They represent a breakthrough in the assessment and management of food safety based on scientific evidence.

Insulin resistance is a prevalent condition commonly associated with unhealthy lifestyles. It affects several metabolic pathways, increasing risk of abnormalities at different organ levels. Thus, diverse medical specialties should be involved in its diagnosis and treatment. With the purpose of unifying criteria about this condition, a scientific-based consensus was elaborated. A questionnaire including the most important topics such as cardio-metabolic risk, non-alcoholic fatty liver disease and polycystic ovary syndrome, was designed and sent to national experts. When no agreement among them was achieved, the Delphi methodology was applied. The main conclusions reached are that clinical findings are critical for the diagnosis of insulin resistance, not being necessary blood testing. Acquisition of a healthy lifestyle is the most important therapeutic tool. Insulin-sensitizing drugs should be prescribed to individuals at high risk of disease according to clinically validated outcomes. There are specific recommendations for pregnant women, children, adolescents and older people.

Desarrollar e intentar alcanzar una elevada calidad en el servicio prestado por los profesionales de enfermería de los centros geriátricos mediante la implantación de sistemas de gestión de calidad es tema de actualidad, hecho este probado ya en otros trabajos. Es por esta razón que el objetivo general de este estudio se haya centrado en analizar los efectos más relevantes que surten de la instauración de una forma de gestionar la calidad basada en la Norma UNE EN ISO 9001:2008 en el área de enfermería de cuatro residencias de ancianos. Se ha llevado a cabo un análisis de más de 200 encuestas de satisfacción respecto del servicio de enfermería de cuatro residencias de personas mayores, así como una revisión bibliográfica de los resultados obtenidos tras la implantación de un sistema de gestión de calidad en el área de enfermería de residencias geriátricas. Apreciamos resultados muy positivos en el desarrollo diario de las funciones de enfermería de las residencias geriátricas derivados de la utilización de un sistema de gestión de calidad, pues ha mejorado considerablemente la calidad en la práctica clínica. El estudio muestra que la implantación de un sistema de gestión de calidad proporciona una herramienta útil al servicio de enfermería en residencias de personas mayores evidenciándose los efectos positivos que resultan de dicha implantación mediante encuestas de satisfacción e indicadores sanitarios.

Introduction Type 1 diabetes (T1D) has a complex etiology in which genetic and environmental factors are involved, whose interactions have not yet been completely clarified. In this context, the role in PD-1 pathway and its ligands 1 and 2 (PD-L1 and PD-L2) have been proposed as candidates in several autoimmune diseases. The aim of this work was to determine the allele and haplotype frequency of six gene polymorphisms of PD-ligands (PD-L1 and PD-L2) in Chilean T1D patients and their effect on serum levels of PD-L1 and autoantibody profile (GAD65 and IA2). Methods This study cohort comprised 205 T1D patients and 205 normal children. We performed genotypic analysis of PD-L1 and PD-L2 genes by TaqMan method. Determination of anti-GAD65 and anti-IA-2 autoantibodies was performed by ELISA. The PD-L1 serum levels were measured. Results The allelic distribution of PD-L1 variants (rs2297137 and rs4143815) showed differences between T1D patients and controls (p = 0.035 and p = 0.022, respectively). No differences were detected among the PD-L2 polymorphisms, and only the rs16923189 showed genetic variation. T1D patients showed decreased serum levels of PD-L1 compared to controls: 1.42 [0.23–7.45] ng/mL versus 3.35 [0.49–5.89] ng/mL (p

Developing and trying to achieve a high quality in the service nursing personnel provide at nursing homes by means of implementing quality assurance systems is a topic of general interest nowadays. These systems have already proved to be effective in other fields. That is why the focus of this study is to analyse the most relevant effects due to the implementation of a quality assurance method based on standard UNE EN ISO 9001:2008 in the nursing service of four nursing homes. An analysis of more than 200 answers of a satisfaction survey about the nursing service at four nursing homes has been carried out. A study on the available bibliography about the results obtained after having implemented a quality assurance system in the nursing service at nursing homes has also been carried out. Very positive results can be appreciated in the daily nursing practice at nursing homes as a consequence of using a quality assurance system. Clinical practice has improved considerably as a consequence. The study shows quality assurance systems to be a useful tool at the service of nursing practice at nursing homes. Positive effects resulting from the implementation of such quality assurance systems are evident as it can be seen from satisfaction surveys and health indicators.

Background Pollution and viral infections could be associated with the incidence of type 1 diabetes mellitus. Aim To look for associations between the temporal patterns of Type 1 Diabetes Mellitus (T1D) in infants younger than the age of 15 years, and environmental factors, such as air pollution and viruses. Material and methods Data registries from hospitals, emergency services, and the Infantile Diabetes Foundation were reviewed, corresponding to children aged less than 15 years, who received their first insulin injection between 2000 and 2007. The incidence of type 1 diabetes was computed for each epidemiological week. Environmental ozone and particulate matter rates for each week were obtained from Environmental services. Rates of influenza and respiratory syncytial virus infections were obtained from the epidemiological department of the Ministry of Health. An ecological Bayesian Poisson regression model was fitted, introducing the covariates, lagged covariates and errors, to estimate the incidence by epidemiological week. Results Three factors were significant by the proposed model: particulate matter PPM 2.5 (relative risk (RR): 1.003) lagged by two weeks, influenza (RR: 0.1808) and RSV (RR: 1.021). Trends and seasonality were clearly controlled by these covariates, considering the epidemiological week as a counting period. Conclusions These results show that environmental factors could be related to peaks of type 1 diabetes incidence.

Purpose There is increasing evidence on the relationship between endogenously produced immunoglobulins and the clinical outcome in septic shock (SS). Materials and methods Levels of immunoglobulin G (IgG) subclasses, immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin E were measured in plasma from 42 patients with SS and in 36 patients with systemic inflammatory response syndrome at diagnosis. Association of immunoglobulins levels with disease severity and outcome was evaluated. Results Eighteen patients with SS finally died. Both patients with systemic inflammatory response syndrome and SS showed subnormal levels of total IgG, IgG2, and IgM. Patients with SS who died showed the lowest levels of total IgG and IgG1. Total IgG, IgG1, IgG2, IgG3, IgG4, and IgA correlated inversely with Acute Physiology and Chronic Health Evaluation II score in SS. Univariate Cox regression analysis showed that levels of IgG1, IgG2, IgG3, IgM, IgA, and total IgG were inversely associated to the probability of death at 28 days. Multivariate analysis showed that IgG1, total IgG, IgM, and IgA behaved as independent protective factors against mortality (hazard ratio, P ): 0.23, 0.026; 0.16, 0.028; 0.11, 0.042; 0.05, 0.010, respectively, whereas IgG3 showed a protective trend also. Conclusions Our study evidenced that, in addition to IgG1, other major endogenous immunoglobulins isotypes and subclasses seem to play a beneficial role in SS.

“Hurdle technology” is a worldwide technique of food preservation based on the application of a combination of generally mild treatments which act as “obstacles” that microflora must overcome to start to grow. Then, bacteria invest their energy in trying to maintain their homeostatic equilibrium instead of multiplying. While the action mechanisms underlying these treatments are not fully understood, it is very useful to know their effect on bacteria cells as well as the extension of such effects. Growth/no growth models have been developed to offer a response to this need. A review on growth/no growth microbial modeling is presented in this paper, addressing the most important factors and approaches employed. Five growth/no growth models of Salmonella published in the period 2001–2011 are reviewed, and their boundary regions were represented (Temperature vs pH) at two water activity values (0.983 and 0.990) and two cut-off probabilities (0.1 and 0.5). With this illustration, a picture of the relative grade of conservatism of the five models is provided. Additionally, the most important predictive tools in food microbiology (or tertiary models), including a software for growth/no growth modeling (Microbial Responses Viewer), are commented. Finally, some caveats in growth/no growth Salmonella modeling are addressed for future research.

The presence of Salmonella in foodstuffs represents an internationally accepted human health concern. Although Salmonella causes many foodborne disease outbreaks, there is little evidence to support cross-contamination as a major contributing factor. However, the paramount importance of preventing cross-contamination and recontamination in assuring the safety of foodstuffs is well known. Sources and factors linked to cross-contamination and recontamination of Salmonella in foods are reviewed in detail. Those foods which are not submitted to lethal treatment at the end of processing or which do not receive further treatment in the home deserves special attention. Salmonella cross-contamination and recontamination episodes have been connected to the following factors: poor sanitation practices, poor equipment design, and deficient control of ingredients. We also examine potential cross-contamination in the home. Cross-contamination and recontamination events at factory level evidence the difficulty encountered for eradicating this pathogen from the environment and facilities, highlighting the need to reinforce industry preventive control measures such as appropriate and standardized sanitation. Also, at consumer level, Public Health Authorities should install hygiene education programs in order to raise consumer awareness of the risks of cross-contamination in the home and their role in its prevention. Finally, a review on cross-contamination models of Salmonella spp. is presented.

Severe sepsis is a common occurrence in critically ill patients and a major cause of morbidity and mortality in this population. Management relies on the early identification and treatment of the underlying causative infection, adequate and rapid hemodynamic resuscitation, support of associated organ failure and modulation of the immune response with drotrecogin alfa (activated) when it is not contraindicated, and corticosteroids in severe septic shock. We will review current approaches to each of these categories.

Chilled Ready-To-Eat (RTE) foods are of concern in hospital foodservices because they can support microbial growth when subjecting to time/temperature abuses during processing and distribution together with poor handling practices. This study was conducted in five different hospitals (A–E) of Southern Spain during 2008–2009 to perform an evaluation of their sanitary conditions and microbiological quality of two RTE meals: lettuce salads and cooked ham. A checklist based on hygiene principles embedded in Food European legislation was developed and applied in each hospital. In parallel, microbiological analysis of food contact surfaces, air quality and time/temperature measurements along the distribution chain were carried out. RTE samples (n = 150) were examined for mesophilic aerobic bacteria (MAB), total coliforms, coagulase-positive Staphylococci (CPS), Escherichia coli, Listeria spp. and Salmonella spp. Differences were found between hospitals regarding handling practices and cleanliness of working surfaces. Cooked ham samples presented lower counts of MAB and total coliforms (

Introduction: Deregulation of apoptosis across the Fas–FasL pathway is an increasingly relevant phenomenon in the pathogenic mechanisms associated with autoimmune diseases. Caspase-8 initiates the activation of the apoptotic process and interacts directly with Fas in the membrane of the T lymphocyte. Objectives: To standardize an Elisa essay to measure the concentration of anti-caspase-8 antibodies in plasma of Type 1 Diabetes (T1D) patients and analyze their possible distribution and association with characteristics of the disease. Methods and subjects: 124 patients newly diagnosed with T1D and 132 controls: children and youngsters. ELISA test was standardized to detect anti-caspase-8 antibodies in plasma. It correlated the concentration of this antibody with classical markers of autoimmunity as anti-IA-2 and anti-GAD65, and the clinical characteristics at onset of diabetes mellitus. The statistical analysis was performed using logistic regression. Results: Patients with T1D showed a higher concentration of anti-caspase-8 antibodies regarding the controls (87.5 ng/ml versus 24.3 ng/ml, p < 0.0001, values expressed as median). The proportion of patients with T1D and high concentrations of anti-caspase-8 (percentile 50–75) was significantly different from the control group (p < 0.0001). Anti-caspase-8 showed a strong association with positive anti-GAD65 (OR = 3.48, p < 0.035) and ketoacidosis (OR = 10.74, p < 0.0001) events, with glycemia and age at diagnosis as contributing variables. Conclusion: This is the first report in the literature of levels of anti-caspase-8 antibodies in T1D through ELISA. The high concentration in patients with T1D, and its strong correlation with anti-GAD65 autoantibodies, suggests a potential role of anti-caspase-8 auto-antibodies as surrogate marker autoimmunity in T1D patients. © 2010 Elsevier GmbH. All rights reserved.

To explore the contribution of the PD-1 gene polymorphisms involved in T1D as well as the relationship between the PD-1/CTLA-4 genes and soluble CTLA-4 concentrations.261 incident cases of T1D and 280 healthy children less 15 years old were included in this study. Haplotypes for polymorphisms of the PD-1 and CTLA-4 genes were determined by PCR and RFLP methods. Screening for soluble CTLA-4 was done using an ELISA assay. Statistical analysis was performed using the online SHESIS package.Our results show that sCTLA-4 levels were higher in T1D than in controls (2.99+/-1.7 ng/ml versus 1.43+/-0.31 ng/ml, p0.001). The allele dosage of CTLA-4 on PD-1 haplotypes, showing a significant modified effect of G carriers over AA genotype on the sCTLA-4 concentrations (5.48+/-2.09 ng/ml versus 3.27+/-1.30 ng/ml, p0.03 in T-C haplotype) and (1.92+/-0.79 ng/ml versus 3.41+/-1.10 ng/ml, p0.02 in C-T haplotype).Consistent with the higher serum sCTLA-4 levels observed in other autoimmune diseases, our results suggest that sCTLA-4 is elevated in T1D. Our data suggest a possible gene dosage effect of "G"CTLA-4 carriers on sCTLA-4 over the possible protective or susceptible effect conferred by PD-1 haplotypes.

Bacterial transfer events are considered to be an important cause of transmission of food-borne diseases. In this review the most common ways of bacterial transfer to food are studied, concluding that high levels of moisture, contact time and pressure could result in higher transfer between surfaces. From a risk management approach, this means that the identification and elimination of certain tasks or activities involving higher levels of these factors could be helpful to prevent bacterial transfer. Besides, mathematical bacterial transfer models from literature are analysed, highlighting the use of transfer rates distributions to model bacterial transfer in a probabilistic Quantitative Microbiological Risk Assessment (QMRA) framework.

Fundamento y Objetivo Analizar el polimorfismo Fok I del gen del receptor de vitamina D (VDR) y su relacion con la susceptibilidad a desarrollar diabetes tipo 1. Se realizo un estudio de casos y controles y se relacionaron los genotipos del VDR con las concentraciones circulantes de 25(OH)-vitamina D y las concentraciones sericas de factor transformador de crecimiento β1 (TGF-β1) e interferon gamma (INF-γ). Pacientes y metodo Se analizaron 151 casos de diabetes tipo 1 y 182 controles sanos no relacionados. Se amplifico el exon 2 del gen VDR mediante reaccion en cadena de la polimerasa. Los genotipos se obtuvieron mediante analisis de fragmentos de restriccion. La concentracion de 25(OH) vitamina D se determino por radioinmunoanalisis y las de las citocinas TGF-β1 e INF-γ, por enzimoinmunoanalisis. Resultados La distribucion del polimorfismo Fok I no mostro diferencias entre casos y controles. El grupo de ninos diabeticos mostro valores mas altos de TGF-β1 (mediana, 282,6 pg/ml; intervalos, 131,8-3.031,4) que los ninos controles (mediana, 232,2 pg/ml; intervalos, 135,7-506,5) (p Conclusiones El polimorfismo Fok I del VDR podria tener un papel marginal en la alteracion inmuno-logica que se desarrolla en la diabetes tipo 1 a traves de una posible modulacion de la vitamina D.

Objective To evaluate the influence of ApaI, BsmI and TaqI polymorphisms of the VDR gene and HLA-DQB1* alleles in type 1 diabetic children and to assess their possible relationship with circulating levels of 25-hydroxyvitamin D3, auto-antibodies, and INFγ/TGFβ1 cytokines levels in Chilean cases and controls. Methods DNA and serum samples from 216 newly diagnosed type 1 diabetic and 203 unrelated control children were evaluated for IA-2 and GAD65 auto-antibodies, 25-hydroxyvitamin D3 levels, HLA-DQB1* alleles, and VDR gene polymorphisms. Results The frequency of the b allele and the bb genotype in type 1 diabetic patients was significantly lower compared with the control group (0.635 versus 0.749, p

Within the elderly population, the specific circumstances that some of the elderly find themselves due to certain socio-cultural factors make them more likely to be abused while residing in assisted living residences, due to the facts that the abuse is coming from the organisation it is unlikely to be reported, or brought to the attention of others, making it difficult to detect and, as a result, an issue not often discussed. We intend to investigate these possibilities; however, due to lack of information on this subject, we aim to address this issue by investigating the number of elderly people living in nursing homes who have suffered abuse due to agents outside of the organisation, such as the family, other residents and Friends. A brief questionnaire was created on an Excel Spreadsheet for the collection of data for this study, these questionnaires where then passed on the social workers of 4 assisted living residences, in the geographical scope of the Murcia region. The data collected correspond to the time period between January and December 2013. The results lead us to believe that the principal risk factors associated with this type of abuse is being a woman, a widow and with and age of about 80 years. Psychological abuse from family members was the most frequently reported type of abuse in the study.

The aim of this study was to estimate the prevalence of type 2 diabetes mellitus (DM2), impaired glucose tolerance (IGT), and the frequency of dyslipidemia, obesity, and hypertension in the rural Aymara population from Northern Chile. In this cross-sectional study, 196 Aymara adult subjects were characterized with respect to their reported physical activity, fasting plasma glucose levels, insulin concentrations, blood pressures, body mass indexes, and plasma lipid profiles. The participants also underwent a 2-h oral glucose tolerance test. The diagnostic criteria for DM2 and IGT followed those of the World Health Organization. The overall prevalence of DM2 was estimated as 1.5% (95% confidence interval: 0.3--4.5). Overall prevalence of IGT was calculated as 3.6% (1.5--7.3). The occurrence of obesity and dyslipidemia was relatively high in the Aymara population, although the frequency of sedentary habits, and the prevalence of hypertension were low. In conclusion, the prevalence of DM2 in the rural Aymara population living at high altitudes in Northern Chile, was much lower than that of other Amerindian groups that adopted lifestyles from industrialized Western societies. Despite a relatively high prevalence of a body mass index of at least 30 kg/m(2), especially in women (23.5%), high physical activity levels and low plasma-insulin concentrations may have been responsible in part for the low prevalence of DM2 in the Aymara population.

The purpose of this study was to examine the relationship of plasma leptin levels with respect to obesity, gender, age and insulin levels in the native Aymara population. The Aymara natives live at high altitudes in isolated regions in the north of Chile, and they maintain distinctive genetic and cultural characteristics. Plasma leptin and insulin levels were correlated with body mass index (BMI), sex and age in a sample of 147 adult Aymara subjects who participated in a cross-sectional study. Multivariate analysis showed significant differences in leptin levels (dependent variable: natural log of leptin) by gender (p < 0.0001), and by BMI (p < 0.001), without significant statistical interaction between gender and BMI. The effect of age achieved statistical significance in the multivariate analysis (p = 0.02). Gender, BMI and insulin are independently associated with plasma leptin levels. On the other hand, the multivariate analysis of the plasma insulin concentration (dependent variable: natural log of insulin levels) shows that insulin is strongly associated with BMI (p < 0.0001), although non-statistically significant differences of insulin levels by sex (p = 0.07) or age (p = 0.9) were detected at alpha 0.05 level. Thus, in the special ecosystem where the Aymara population live, a strong and independent association between sex, obesity and insulin levels with plasma leptin levels has been detected.

CTLA-4 plays a key role in T cells regulation. We analysed the CTLA-4 +49A/G and −318C/T polymorphisms in 178 cases of type 1 diabetes and their parents (534 individuals) from Santiago, Chile. A significant overall association with T1D ( p = 0.028) was observed, possibly due to an overtransmission of the G–T haplotype.

In case-control studies, spurious associatons between Human Leukocyte Antigen (HLA) alleles and Type 1 diabetes could arise as a result of population stratification, if there are ethnic differences between cases and non-related controls. The Chilean population has several unique features which make it ideal for the study of the effect of stratification by ethnicity on genetic epidemiological research. The incidence rates of Type 1 diabetes in Chilean Aboriginal populations are very low compared to Caucasian populations, while the frequency of the alleles in HLA loci also vary across ethnic groups. In order to avoid the confounding effect of ethnicity, one possible remedy would be the use of cases and their parents in place of non-related controls. The case-parent design offers an adequate framework for the study of the association between HLA polymorphisms and Type 1 diabetes in the Chilean population and can also be applicable to other genetically mixed populations especially in the Americas.

The establishment of validated methodologies for the determination of food shelf-life is currently demanded by both food industries and Health Authorities at national and international scale. It is well known that most foods are perishable, since they are subjected to modifications in their structure, composition and properties during storage before consumption. These changes are of physico-chemical origin attributed to food composition together with the action of intrinsic and extrinsic environmental factors, and also microbiological, where spoilage flora play an important role. These modifications are “translated into“ sensorial deterioration at a specific time point. In this respect, food-borne bacteria, despite representing a threat for consumers health, do not affect sensorial changes.

It is a principal aim of governments to assure the safety of societies in all sectors. In the food field, safety has been dealt with for a long time by making decisions in an empirical manner. Recently, risk management has been appointed as the formal scientific-based approach to address food safety issues. From a global perspective, food safety risk management can be described “as the process of weighting control alternatives by government (and international standard-setting bodies) in consultation with interested stakeholders, taking into account scientific information on risks to consumers as well as other relevant inputs (e.g. economics, technical feasibility, societal preferences), and choosing and implementing food safety measures as appropriate” (Food Agriculture Organization/World Health Organization [FAO/WHO], 2006b). Indeed, governments must make decisions, whose effects are especially noted during food crises. Nevertheless, other stakeholders should also manage food risks, for example, at manufacture or consumer level. Nowadays, manufacturers and other operators involved in the food chain are aware of the importance of producing and assuring food safety, as well as the devastating consequences of supplying contaminated food products. However, at consumer level, the relevance of consumer s hygiene practices in the home may not always be evident in order to avoid foodborne conditions. Food safety risk management should be based on risk assessment, as proposed by the Regulation (EC) No. 178/2002 of 28 January 2002, laying down the general principles and requirements of food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety. Currently, risk assessment is being gradually introduced at governmental level as a systematic practice. In the case of manufacturers and other food business operators, only big food enterprises have adopted risk assessment procedures; risk management in medium and small food companies is based on the implementation of Hazard Analysis and Critical Control Point (HACCP) systems, which is in fact compulsory. Hygiene practices in homes are out of the control of Health Authorities; nevertheless, for example, a risk management option mandated by governments may consist of developing educational programs which could enhance a positive attitude among consumers towards a more hygienic preparation of foods. Food safety risk management has been proved to be useful in making science-based decisions. In this chapter, food safety risk management is addressed from various perspectives, together with management metrics to facilitate its implementation. Also, a review of risk assessment is included. A promising future can be envisaged for food safety risk management activities.

Food safety measures in long-term care facilities (LTCFs) are being improved by the introduction of quality control management systems during food production and by the implementation of good manufacturing practices. This study was conducted in LTCFs (geriatric homes) in Andalusia, Spain, during 2008 and 2009 to evaluate sanitary conditions and the microbiological quality and safety of salads and cooked meat products served. A regulation-based checklist was applied to the evaluated centers. Samples of ready-to-eat foods (n = 60) were examined for mesophilic aerobic bacteria (MAB), total coliforms, coagulase-positive staphylococci (CPS), Escherichia coli, Listeria spp., and Salmonella. In parallel, food contact surfaces (working tables, cutting boards, sinks, and faucets) were swabbed and analyzed for MAB and Enterobacteriaceae. The air quality in processing rooms, near sinks, and in canteens also was measured through an active air sampling method for MAB and Staphyloccocus spp. The results obtained revealed some deficiencies regarding handling practices and sanitary conditions tested (i.e., use and change of gloves, hand washing, and cleanliness of work surfaces). The microbial safety of foods examined indicated the absence of pathogens. Average levels of coagulase-positive staphylococci were below 10² CFU/g, and prevalence of E. coli was 6.3% in samples collected. Surface counts were higher on cutting boards and faucets, indicating insufficient cleanliness procedures. This study provides a descriptive analysis of the sanitary conditions of food service systems in LTCF, and this information can help risk managers to better define control measures needed to prevent foodborne infections.

The relationships between cytokine responses in septic shock are currently poorly understood. Some studies have pointed to a biphasic model, with an initial proinflammatory phase, followed by a reactive, anti-inflammatory response to explain the pathogenesis of the most severe form of sepsis. However, evidence for the coexistence of both responses has been found. In this study, the plasma levels of 17 cytokines and chemokines, in 20 patients with septic shock, 11 patients with systemic inflammatory response syndrome (SIRS), during the first 24 hours following diagnosis, and 10 healthy controls, were analyzed and compared. Patients with septic shock showed increased levels of IL-6, IL-8, MCP-1, MIP-1β, IFN-γ, GM-CSF and IL-10 compared to healthy controls. Patients with SIRS showed higher levels of IL-6, IL-8, MCP-1, MIP-1β, G-CSF and IL-10 than controls. Patients with septic shock showed higher levels of IL-8, GM-CSF, MIP-1β than those with SIRS. The Spearman test demonstrated a positive association between the pro-inflammatory mediators IL-6, IL-8, MCP-1, MIP-1β, IFN-γ, GM-CSF and the immunomodulatory cytokine IL-10 in septic shock. Consequently, correlation studies supported the notion that secretion of pro- and anti-inflammatory mediators in septic shock occurs as a simultaneous immune response program initiated early in the course of the disease, revealing that both types of cytokine play a role from the very beginning of this life-threatening condition.

Gestational diabetes mellitus (GDM) is highly prevalent in women with polycystic ovary syndrome (PCOS). Women with GDM have considerable risk for developing both type 1 and type 2 diabetes.To evaluate the prevalence of anti-GAD65 and anti-IA2 auto-antibodies in Chilean pregnant women with GDM, normal pregnancy (NP) and with PCOS (PPCOS) to establish whether in PCOS women GDM is partially induced by auto-antibodies.Women with singleton pregnancies matched by age and gestational age were included: 50 GDM, 59 NP and 50 PPCOS. During gestational weeks 22-28, a 2-h, 75 g oral glucose tolerance test was performed, with measurement of glucose, insulin, lipids and auto-antibodies.A highly prevalence of anti-GAD65 antibodies (12%) was observed in women with GDM. PPCOS and NP women showed a similar distribution of anti-GAD65 antibodies (2.0% and 1.7%, respectively). Anti-IA2 antibodies were present in 4.0% of women with GDM, in 1.7% of NP women and 2.0% PPCOS women.A highly prevalence of anti-GAD65 was observed in women with GDM which is in agreement with previous studies. Nevertheless, the frequency of these auto-antibodies was very low in NP and PPCOS women.

A practical approach was used to achieve specified food safety objectives (FSOs) by incorporating risk assessment methodology. A series of cross-contamination scenarios at the retail level were evaluated in a spreadsheet and ranked according to their level of risk, and the degree to which they exceeded an FSO for Listeria monocytogenes (100 CFU/g) was determined. Percentiles and analyses of variance were used to identify which scenarios would be most likely not to achieve the FSO. It was also possible to incorporate into the model the effect of additional risk management interventions (e.g., hand washing method) by using "what if" scenarios analysis. The highest risk corresponded to the use of the same gloves to handle contaminated meat and then sliced ham compared with the safer method of using different gloves to handle each product. Nevertheless, in all scenarios bacterial concentrations could be higher than 10(8) CFU/g. A significant lack of knowledge regarding the transfer rates of L. monocytogenes was revealed, which provided the model with an important uncertainty component. The "what if" scenarios analysis based on the scenario representing handling food with bare hands followed by hand washing (scenario 2) revealed that the hand washing procedure had less impact than expected. This study illustrates the use of scenarios to model the most effective food processes and thus provide risk managers with appropriate information on which to base their decisions.

Background: There are great geographical differences in the incidence of type I diabetes mellitus. Aim: To determine the incidence rate of type 1 diabetes mellitus (DM1) in the Metropolitan Region of Santiago, Chile from January 1, 2000 to December 31, 2004 and to observe the distribution of cases in the different counties of Santiago. Material and methods: All the cases diagnosed with DM1 in the Metropolitan Region who fulfilled the following requirements were included in the study: age of onset

Study objective and setting The aim of this study is to determine the incidence rate of type 1 diabetes in the metropolitan region of Santiago, Chile from 1 January 1986 to 31 December 2003. Design Population-based incidence study. Participants: A case must fulfill the following requirements to be included in this study: age at onset: 0 to 14 years, diagnosed with diabetes and placed on insulin, diagnosed within the defined time period, and to be a resident of the metropolitan region of Santiago at the time of the diagnosis. The population ‘at risk’ is the population less than 15 years of age. Result The overall rate of type 1 diabetes was estimated as 4.02 cases per 100 000 children per year (95% confidence interval: 2.98–4.83). Conclusion The incidence of type 1 diabetes in Santiago, Chile has increased during the last years. This data are concordant with the observation that the incidence of type 1 diabetes is increasing in Latin America and worldwide. Copyright © 2005 John Wiley & Sons, Ltd.

0.41 yr). Incidence of GDMwas 14.4% among women 25 years old or older and increased to 21.4% when they had, in addi-tion, a BMI of 25 or over. Age, BMI, and family history of diabetes were all independently correlat-ed with the development of GDM. Elective caesarean sections were more common in GDM than innon-GDM women (p =

Background: Type 1 diabetes is an organ specifc autoimmune disease whose incidence is increasing worldwide. A functional imbalance in cytokine production resulting in dominance of T helper (Th1) over Th2-type response has been suggested to play a critical role in the pathogenesis of type 1 diabetes. Aim: To measure serum concentrations of interleukin (IL)-1ß, IL-2 and IL-4 in children with recently diagnosed type 1 diabetes and to evaluate the autoimmune response measuring glutamic acid decarboxylase (GAD65) and tyrosine phosphatase like (IA-2) autoantibodies. Patients and Methods: 120 diabetic children and 118 age and gender matched control children, were recruited for this study. Circulating levels of IL-1ß, IL-2 and IL-4 were measured by ELISA. GAD65 and IA-2 were measured by RIA. Results: Circulating levels of IL-1ß were elevated in type 1 diabetic children as compared to the control group (9.3±7.3 and 4.9±3.8 pg/ml respectively, p=0,01). Serum concentration of IL-2 was also higher in diabetic patients (19.8±13.1 and 11.3±9.1 pg/ml respectively, p=0,01). No differences in serum IL-4 were observed between diabetics and control. Diabetic children with one or two positive autoantibodies (IA-2 and/or GAD65) had significantly higher levels of IL-1ß and IL-2 and lower levels of IL-4 than diabetic children without positive autoantibodies. High concentrations of IL-1ß were associated with an early onset of the disease. Conclusions: High levels of IL-1ß and IL-2 were found in diabetic children with recent diagnosis of the disease. Diabetics with positive antibodies against GAD65 and IA-2 had higher levels of IL-1ß and IL-2 and lower levels of IL-4 than their counterparts without positive antibodies (Rev Méd Chile 2004; 132: 413-20)

Objective: To compare the levels of bovine serum albumin (BSA) antibodies and their relationship with duration of breast feeding, age of exposure to cow's milk, and human leukocyte antigen (HLA-DQ) genotype in children with and without type 1 diabetes. Methods: Serum samples from 143 (0.3–14.7 yr) newly diagnosed children with type 1 diabetes and 107 unrelated control children (0.8–13.5 yr) were evaluated for BSA antibodies. Duration of breast feeding and exposure to cow's milk were recorded on questionnaires. HLA-DQ typing was determined by polymerase chain reaction. Results: One hundred percent of the diabetic children were positive for BSA antibodies compared to 1.9% for healthy controls (p

Several reports have found a relation between polymorphisms of the vitamin D receptor gene (VDR) and the development of type 1 diabetes. We have examined the association of three VDR polymorphism with type 1 diabetes in 59 Chilean case-parents trios. Genotyping for Bsm1, Apal and Taq1 polymorphism were performed. Transmission/ disequiibrium tests were used to assess gene-disease associations through the evaluation of allelic transmission to affected offspring. Non-significant increased transmissions of B allele (probability of transmission = 52.5%, p = 0.69), A allele (probability of transmission = 58.4%, p = 0.17) and T allele (probability of transmission = 52.0%, p = 0.77) were estimated in Bsml, Apal and Taql sites, respectively. Haplotype-based analyses showed non-significant preferential transmissions (global p = 0.52). The present study does not support the hypothesis of a significant contribution of VDR alleles in the etiology of type 1 diabetes of Chilean cases.

The aim of this study was to estimate annual incidence rate of type 1 diabetes according to the levels of genetic susceptibility provided by HLA-DQA1 and HLA-DQB1 genotypes. Two information sources were used: (1) a population-based incidence study in which 61 incident cases were ascertained during 1 year in Santiago, Chile (incidence rate: 4.11 cases per 100,000 children per year) and (2) a case-control comparison of 57 cases (recruited from the incidence study) and 125 controls. Susceptibility alleles were defined as DQA1*0301 and DQA1*0501 for DQA1 gene and alleles DQB1*0201 and DQB1*0302 for DQB1 gene. In DQA1 gene, the highest point estimate of the incidence rate was calculated for the genotype DQA1*0501/DQA1*0501 (33.04 cases per 100,000 children aged less than 15 years old and per year; 95% CI: 9.22-118.33). In the DQB1 gene, the highest risk was estimated for the genotype DQB1*0201/DQB1*0201 (20.35 cases per 100,000 children aged less than 15 years old and per year; 95% CI: 5.26-78.67). This study shows an application on how a transformation of the logistic equation based on Bayes' theorem can be used to estimate incidence rates from case-control studies and population-based incidence rates.

Background: Environmental and genetic factors (viruses, toxins and diet) are involved in the aetiology of type 1 diabetes. Among the dietary factors, the role of breast feeding and exposure to cow’s milk proteins deserve special attention. Aim: To determine the anti-BSA-IgG levels in type 1 diabetic children and to analyse the possible association with breast feeding duration, exposure to cow’s milk and s pancreatic auto-antibodies. Patients and methods: Blood samples were collected from 161 diabetic children and 144 controls to measure anti-BSA-IgG level, GAD65, IA-2 and ICA autoantibodies. All children answered a questionnaire about dietary habits during infancy. Results: anti-BSA-IgG was positive (using a cut off point of 25.6 ng/ml) in 98% of diabetic children and 0% of the control population. The length of breast feeding or early exposure to cow’s milk did not influence the concentration of anti-BSA-IgG. Positive BSA titers did not increase the s pancreatic reactivity (ICA+, GAD+, IA2+). Conclusions: Our data confirm the high frequency of anti-BSA-IgG among diabetic children. However, a specific role in the immunological process of type 1 diabetes cannot be attributed to this protein (Rev Med Chile 2003; 131: 865-72). (Key Words: Breast feeding; Diabetes Mellitus, insulin-dependent; Insulin antibodies; Milk proteins)

Background: Islet cell-specific autoantibodies such as islet cell antibody (ICA), antiinsulin (IAA), anti-glutamic acid decarboxylase (GAD) and anti-tyrosine phosphatase (IA2) can be present in patients with type I diabetes. Breast feeding duration and the early exposure to milk substitutes are environmental factors associated to etiology of type 1 diabetes. Aim To study the frequency of the anti-GAD, anti-IA-2 e ICA antibodies in Chilean type 1 diabetic patients and determine the possible modulator effect of the breast feeding. Patients and methods: One hundred thirty four type I diabetic patients, aged one to 15 years old, were studied at the moment of their diagnosis. Patients were classified according to the duration of exclusive breast feeding. IA-2 and GAD were determined by radio immuno assay and ICA by means of indirect immunofluorescence. Results: Subjects with three months or less and those with more than three months of breast feeding were positive for ICA in 78.8 and 90.6% of cases respectively, for GAD in 75 and 54.6% of cases respectively (p=0.024) and for IA-2 in 73 and 43.8% of cases respectively (p=0.001). All three antibodies were positive in 53.9 and 21.8% of children with less or more than three months of breast feeding (p=0.001). Conclusion: Both IA-2 and GAD antibodies are less frequently positive in type 1 diabetic patients who have been breast fed for more than three months. These findings suggest a possible attenuating role of exclusive breast feeding on pancreatic aggression events in patients with type 1 diabetes (Rev Med Chile, 2001; 129: 611-9)

The aim of this study was to estimate the prevalence of Type 2 diabetes, impaired glucose tolerance (IGT), and obesity in the Mapuche natives from rural areas in Chile. This cross-sectional study involved men (n = 95) and women (n = 224) older than 20 y from an aboriginal ethnic group (Mapuches), residing in rural communities from the south of Chile. Prevalence of Type 2 diabetes and IGT was calculated according to the World Health Organization criteria. Data on age, degree of ancestral purity, obesity, and hypertension were also obtained. The prevalence of Type 2 diabetes in rural Mapuche natives was estimated as 3.2% (95% CI: 0.7--9.0) in men and 4.5% (95% CI: 2.2--8.1) in women. The overall prevalence of obesity was 56.1% (95% CI: 50.5--61.6): 40.0% (95% CI: 30.1--40.8) in men and 62.9% (95% CI: 56.3--69.3) in women (P value0.001). These data suggest that the prevalence of obesity and Type 2 diabetes has been increasing during recent years in the Mapuche communities. The prevalence estimated in this study is higher than that reported 15 y ago. This suggests an important role of lifestyle changes as a possible explanation for epidemiologic transition.

The human leukocyte antigen (HLA) system plays a crucial role in the autoimmune process leading to childhood diabetes. The purpose of this study was to evaluate the association between type 1 diabetes and the polymorphism encoded by the HLA-DQB1 gene by using case-parents trios. The study area was the metropolitan region of Santiago, Chile, and cases were ascertained from March 1997 to August 1998. Genotyping was performed in 94 trios comprising incident cases less than 17 years of age at the time of diagnosis and their parents. The transmission/disequilibrium test was used to detect differential transmission in the HLA-DQB1 locus. The authors found that alleles DQB1(*)0302 and DQB1(*)0201 were strongly associated with the disease. By using 1:3 matched sets of cases-pseudosibs and conditional logistic regression models, allelic relative risks were estimated for DQB1(*)0302 (r = 7.2, 95% confidence interval: 2.8, 18.5) and DQB1(*)0201 (r = 4.7, 95% confidence interval: 1.9, 11.6); DQB1(*)0301 was considered the baseline allele. When case-parents trios were used, alleles DQB1(*)0302 and DQB1(*)0201 were strongly associated with a higher risk of type 1 diabetes in the population of SANTIAGO

Background: Although there is a clear relationship between body mass index and leptin levels, few authors have addressed the possible influence of ethnic factors on these levels. Aim: To measure serum leptin in three different Chilean aboriginal populations. Subjects and methods: Fasting serum leptin and insulin levels were measured by radioimmunoassay in 345 rural mapuche individuals, 247 rural aymara subjects and 162 urban mapuche subjects. A body mass index of 27.5 kg/m2 was used as cutoff point to classify study subjects. Results: Among the three ethnic groups, women had serum leptin levels three times higher than men. In all three ethnic groups, there was a significant association between leptin levels, body mass index and gender (r2= 0.32 and 0.5 p

Background: Chilean aboriginal ethnic groups (mapuche and aymaras) have a very low prevalence rate of type 2 diabetes. The investigation of a possible relationship between this low prevalence of diabetes and obesity, hypertension and serum lipid profiles in both groups is worthwhile. Aim: To study the prevalence of obesity, hypertension and lipid profile in two Chilean aboriginal communities. Subjects and Methods: The prevalence of obesity, hypertension, fasting serum total cholesterol, HDL cholesterol, triglycerides, glucose, insulin, leptin and oral glucose tolerance test were measured in 345 mapuche (106 male) and 247 aymara (100 male) individuals. Results: Sixty three percent of mapuche women, 37.9% of mapuche men, 39.7% of the aymara women and 27.0% of aymara men had a body mass index over 27 kg/m2. Twenty percent of mapuche men, 18.0% of mapuche women, 9.0% of aymara men and 4.8% of the aymara women had high blood pressure values. Serum HDL cholesterol was below 35 mg/dl in 16% of mapuche women, 14% of mapuche men, 25% of the aymara women and 27% of aymara men. No differences in total cholesterol levels were observed between mapuches and aymaras. Conclusion: Mapuche women have higher prevalence of obesity and high blood pressure than aymara women. Low serum HDL cholesterol has a higher prevalence among aymara individuals.

OBJECTIVE: Leptin, the product of the human ob gene is increased in obese individuals, suggesting resistance to its effect. We examined the relationship of serum leptin levels with respect to obesity, gender and insulin levels in two populations with different ethnic compositions in Chile. METHODS: Leptin and insulin levels were determined by radioimmunoassay (RIA) and correlated with body mass index (BMI), gender and ethnic background. 79 Caucasian subjects from Santiago and 65 Mapuche natives from the Araucania region, Chile, were included in this study. RESULTS: Leptin concentrations in obese subjects were significantly increased in both ethnic groups in relation to lean status: Caucasian and Mapuche obese 19.3±11.6 and 10.1±5.8 (P

Producción Científica, Evaluation of host immune response to infection at the molecular level is a promising avenue to obtain diagnostic and prognostic tools for the clinical management of patients with sepsis. A recent report from Cajander and colleagues [1] has shown the potential of HLA-DR mRNA quantification by real-time PCR as a biomarker of immunosuppression in these patients. IgM is the first immunoglobulin produced in response to infection. In a pilot study, we have employed a next generation quantitative PCR method (nanoliter-sized droplet technology paired with digital PCR (ddPCR)) for detecting the early transcriptomic response of IgM in blood from patients with sepsis. Approval for the study protocol for both scientific and ethical aspects was obtained from the Committee for Clinical Research of Hospital Clínico Universitario, Valladolid, Spain. Written informed consent was obtained directly from each patient or a legal surrogate. The target gene transcript was IGHM, which encodes the constant region of the mu heavy chain, which defines the IgM isotype [2]. In blood, the cells producing IgM transcripts are B lymphocytes expressing CD20 [3], which was employed as housekeeping gene., Instituto de Salud Carlos III (grant PI13/02110)

The goal of this study was to estimate the average annual incidence rate of insulin-dependent diabetes mellitus (IDDM) in Santiago as part of a Multinational Project in Childhood Diabetes (Diabetes Mondiale or DiaMond). Incidence was calculated among subjects under 15 years of age, through a retrospective search and confirmation method from 1 January 1986 to 31 December 1992. Hospitals and private offices of endocrinologists and specialists in diabetes were surveyed. A total of 252 registered cases, 118 boys and 134 girls, for an annual incidences of 2.36/100 000 hab.year, which is one of the lowest validated rates in the Americas.

This study was designed to examine the hypothesis that some environmental factors increase the risk for insulin-dependent diabetes mellitus. Data on dietary history was collected from 80 diabetic children from the Santiago de Chile Registry and from 85 nondiabetic control subjects who were comparable in terms of age, sex, and ethnic characteristics. Early exposure was defined as the ingestion of food sources other than maternal milk before 3 months of age. To define genetic susceptibility to insulin-dependent diabetes mellitus each subject was typed in terms of HLA DQA1 and DQB1, and the possible conformation of susceptible heterodimers was considered as a risk marker. Fewer children were exclusively breast fed in the diabetic group than in the control group (21.55 +/- 15.05 vs 33.95 +/- 20.40 weeks, P0.01). In addition, exposure to cow's milk and solid foods occurred earlier in the diabetic group than in the control group (15.90 +/- 10.95 vs 21.15 13.65 and 16.85 +/- 10.25 vs 21.20 +/- 12.35 weeks, P0.05). Our data show that a short duration of breast-feeding and early exposure to cow's milk and solid foods may be important factors in the development of insulin-dependent diabetes mellitus. The high relative risk observed in individuals genetically predisposed indicates an interaction effect between genetic and environmental components.