Acceleration of the DNA methylation clock among lynch syndrome-associated mutation carriers.

BACKGROUND: DNA methylation (DNAm) age metrics have been widely accepted as an epigenetic biomarker for biological aging and disease. The purpose of this study is to assess whether or not individuals carrying Lynch Syndrome-associated mutations are affected in their rate of biological aging, as measured by the epigenetic clock. METHODS: Genome-wide bisulfite DNA sequencing data were generated using DNA from CD4 + T-cells obtained from peripheral blood using 27 patient samples from Lynch syndrome families. Horvath's DNAm age model based on penalized linear regression was applied to estimate DNAm age from patient samples with distinct clinical and genetic characteristics to investigate cancer mutation-related aging effects. RESULTS: Both Lynch mutation carriers and controls exhibited high variability in their estimated DNAm age, but regression analysis showed steeper slope for the Lynch mutation carriers. Remarkably, six Lynch Syndrome-associated mutation carriers showed a strong correlation to the control group, and two sisters carrying Lynch Syndrome-associated mutations, with no significant difference in lifestyle and similar chronological age, were assigned very different DNAm age. CONCLUSIONS: Future studies will be required to explore, in larger patient populations, whether specific epigenetic age acceleration is predictive of time-to-cancer development, treatment response, and survival. Epigenetic clock DNAm metrics may be affected by the presence of cancer mutations in the germline, and thus show promise of potential clinical utility for stratified surveillance strategies based on the relative risk for imminent emergence of tumor lesions in otherwise healthy Lynch Syndrome-associated mutation carriers.

Acciones educativas de la familia con el escolar durante el confinamiento por la COVID-19 / Family educational actions with schoolers during COVID-19 confinement

Introducción: El confinamiento por la COVID-19 ha generado un desafío en la crianza de los escolares. Objetivo: Examinar el desempeño de acciones educativas de la familia con el escolar durante el confinamiento por la COVID-19. Métodos: Investigación descriptiva, muestreo no probabilístico,112 familias. que cumplían aislamiento social en la comunidad Ramón López Peña, San Cristóbal, Artemisa en 2020. Instrumento utilizado: Escala de acciones educativas de la familia con el escolar durante el confinamiento por la COVID-19, construida y validada por los investigadores con alfa de Cronbach= 0,970). Resultados: Las acciones educativas: cumplir los horarios de alimentación y sueño (70,6 por ciento), distribuir los roles en el cuidado del menor (70,6 por ciento), apoyo emocional (70,6 por ciento), apoyar con las teleclases (70,6 por ciento), siempre se realizaron por las familias con desempeño favorable. Las acciones: ayudar a recuperarse de una emoción negativa (72,7 por ciento), alejar al escolar de las preocupaciones y tensiones del hogar (72,7 por ciento), explicar si no entiende los contenidos o buscar ayuda (72,7 por ciento), casi nunca fueron realizadas por las familias con desempeño desfavorable. Conclusiones: La mayoría de las familias presentaron desempeño favorable de acciones educativas. La dimensión Cuidados del menor fue la mejor expresada. Enseñar al escolar las medidas higiénicas de protección; brindar información a los escolares sobre cómo protegerse; enseñar y practicar formas de saludar y de mantenerse conectados con seres queridos y amigos, fueron las acciones educativas más realizadas por ambos grupos de familia. La complejidad del aprendizaje en casa y el manejo afectivo del niño requieren especial atención(AU)

Introduction: COVID-19 confinement has created a challenge in raising schoolers. Objective: Examine the performance of educational actions of the family with the school during the confinement by COVID-19. Methods: Descriptive research, non-probabilistic sampling, 112 families who complied with social isolation in Ramón López Peña community, San Cristobal, Artemisa province in 2020. Instrument used: Scale of educational actions of the family with school children during confinement by COVID-19, built and validated by researchers with Cronbach´s alpha = 0.970. Results: Educational actions as meeting food and sleep schedules (70.6 percent), distributing roles in child care (70.6 percent), emotional support (70.6 percent), supporting with teleclasses (70.6 percent) were always carried out by families with favorable performance. Actions like helping to recover from negative emotions (72.7 percent), keeping the schooler away from household concerns and tensions (72.7 percent), explaining whether they don't understand the contents or looking for help (72.7 percent) were almost never made by families with unfavorable performance. Conclusions: Most families had a favorable performance in educational actions. The Child Care dimension was the best performed one. Teaching the schoolchildren the hygienic protective measures, provide information to schoolchildren on how to protect themselves, teaching and practicing ways to greet and stay connected with loved ones and friends were the most performed educational actions by both family groups. The complexity of home learning and the child's affective management require special attention(AU)

In Patients With Obesity, the Number of Adipose Tissue Mast Cells Is Significantly Lower in Subjects With Type 2 Diabetes.

Type 2 diabetes (T2D) is a rising global health problem mainly caused by obesity and a sedentary lifestyle. In healthy individuals, white adipose tissue (WAT) has a relevant homeostatic role in glucose metabolism, energy storage, and endocrine signaling. Mast cells contribute to these functions promoting WAT angiogenesis and adipogenesis. In patients with T2D, inflammation dramatically impacts WAT functioning, which results in the recruitment of several leukocytes, including monocytes, that enhance this inflammation. Accordingly, the macrophages population rises as the WAT inflammation increases during the T2D status worsening. Since mast cell progenitors cannot arrive at WAT, the amount of WAT mast cells depends on how the new microenvironment affects progenitor and differentiated mast cells. Here, we employed a flow cytometry-based approach to analyze the number of mast cells from omental white adipose tissue (o-WAT) and subcutaneous white adipose tissue (s-WAT) in a cohort of 100 patients with obesity. Additionally, we measured the number of mast cell progenitors in a subcohort of 15 patients. The cohort was divided in three groups: non-T2D, pre-T2D, and T2D. Importantly, patients with T2D have a mild condition (HbA1c <7%). The number of mast cells and mast cell progenitors was lower in patients with T2D in both o-WAT and s-WAT in comparison to subjects from the pre-T2D and non-T2D groups. In the case of mast cells in o-WAT, there were statistically significant differences between non-T2D and T2D groups (p = 0.0031), together with pre-T2D and T2D groups (p=0.0097). However, in s-WAT, the differences are only between non-T2D and T2D groups (p=0.047). These differences have been obtained with patients with a mild T2D condition. Therefore, little changes in T2D status have a huge impact on the number of mast cells in WAT, especially in o-WAT. Due to the importance of mast cells in WAT physiology, their decrease can reduce the capacity of WAT, especially o-WAT, to store lipids and cause hypoxic cell deaths that will trigger inflammation.

Vulnerabilidad al estrés en pacientes adultos hipertensos / Vulnerability to the stress in hypertensive adult patients

RESUMEN Introducción: En la hipertensión, el estrés constituye un factor sicosocial de riesgo. Objetivo: establecer la relación entre vulnerabilidad al estrés con variables sociodemográficas y clínicas en pacientes hipertensos adultos del Policlínico Santa Cruz. Métodos: Estudio descriptivo, transversal. El universo lo conformaron 65 hipertensos, diagnosticados de enero a junio del 2019. La muestra intencional fue de 58 hipertensos. Para la recogida de información se emplearon: la Historia Clínica Individual y familiar y el Test de Vulnerabilidad al Estrés. Resultados: Predominio del 70,7 % de hipertensos en la tercera subetapa de la adultez, supremacía femenina (69,0 %); la mayoría con secundaria básica terminada (36,2 %); casados (44,8 %) y trabajadores estatales (51,7 %); predominaron los fumadores (81 %); prevaleció la hipertensión arterial grado II (69 %) y dieron cumplimiento al tratamiento el 53,4 %. La mayoría presentó vulnerabilidad al estrés (65,5%); mejor representado en la tercera subetapa de la adultez (81,6 %), mujeres (63,2 %), con secundaria básica terminada (39,5 %), casados (42,1 %), trabajador estatal (55,3 %), fumadores (76,3 %), con hipertensión arterial grado II el 76,3 %; cumplieron el tratamiento siempre el 55,3 %; presentaron nivel vulnerable al estrés el 68,4 % de los hipertensos. Existió asociación entre el nivel seriamente vulnerable al estrés y la escolaridad primaria (p= 0,007). La mayoría de los hipertensos presentaron vulnerabilidad al estrés. El nivel vulnerable al estrés predominó; mejor representado en la tercera subetapa de la adultez, en las mujeres, los fumadores, los hipertensos grado II y los que siempre cumplían con el tratamiento. Existió asociación entre el nivel seriamente vulnerable al estrés y la escolaridad primaria.

ABSTRACT Introduction: In the hypertension, the stress constitutes a psychosocial risk factor. Objective: To establish the relation between vulnerability to the stress with socio-demographics and clinical variables in adult hypertensive patients of the General hospital Santa Cruz. Methods: Descriptive, transverse study. The universe was constituted of 65 hypertensive patients, diagnosed from January until June, 2019. The intentional sample was 58 hypertensive patients. For the information collection, it was used: the Individual and familiar Case history and the Test of Vulnerability to the Stress. Results: Predominance 70.7 % of hypertensive patients in the third sub-stage of the adulthood, feminine supremacy (69.0 %); the majority with secondary school finished (36.2 %); married (44.8 %) and state workpeople (51.7 %); there prevailed the smokers (81 %); the arterial hypertension grade II prevailed (69 %) and they fulfilled the treatment 53.4 %. The majority presented vulnerability to the stress (65.5 %); better represented in the third sub-stage of the adulthood (81.6 %), women (63.2 %), with secondary school finished (39.5 %), married (42.1 %), state worker (55.3 %), smokers (76.3 %), with arterial hypertension grade II 76.3 %; they always fulfilled the treatment 55.3 %; they presented vulnerable level to the stress 68.4 % of the hypertensive patients. Association existed between the level seriously vulnerably to the stress and the primary schooling (p = 0.007). Discussion: Most of the hypertensive patients presented vulnerability to the stress. The vulnerable level to the stress prevailed; better represented in the third sub-stage of the adulthood, in the women, the smokers, hypertensive grade II and those who were always fulfilled with the treatment. Association existed between the level seriously vulnerably to the stress and the primary schooling.

ProphTools: general prioritization tools for heterogeneous biological networks.

Background: Networks have been proven effective representations for the analysis of biological data. As such, there exist multiple methods to extract knowledge from biological networks. However, these approaches usually limit their scope to a single biological entity type of interest or they lack the flexibility to analyze user-defined data. Results: We developed ProphTools, a flexible open-source command-line tool that performs prioritization on a heterogeneous network. ProphTools prioritization combines a Flow Propagation algorithm similar to a Random Walk with Restarts and a weighted propagation method. A flexible model for the representation of a heterogeneous network allows the user to define a prioritization problem involving an arbitrary number of entity types and their interconnections. Furthermore, ProphTools provides functionality to perform cross-validation tests, allowing users to select the best network configuration for a given problem. ProphTools core prioritization methodology has already been proven effective in gene-disease prioritization and drug repositioning. Here we make ProphTools available to the scientific community as flexible, open-source software and perform a new proof-of-concept case study on long noncoding RNAs (lncRNAs) to disease prioritization. Conclusions: ProphTools is robust prioritization software that provides the flexibility not present in other state-of-the-art network analysis approaches, enabling researchers to perform prioritization tasks on any user-defined heterogeneous network. Furthermore, the application to lncRNA-disease prioritization shows that ProphTools can reach the performance levels of ad hoc prioritization tools without losing its generality.

DrugNet: network-based drug-disease prioritization by integrating heterogeneous data.

OBJECTIVE: Computational drug repositioning can lead to a considerable reduction in cost and time in any drug development process. Recent approaches have addressed the network-based nature of biological information for performing complex prioritization tasks. In this work, we propose a new methodology based on heterogeneous network prioritization that can aid researchers in the drug repositioning process. METHODS: We have developed DrugNet, a new methodology for drug-disease and disease-drug prioritization. Our approach is based on a network-based prioritization method called ProphNet which has recently been developed by the authors. ProphNet is able to integrate data from complex networks involving a wide range of types of elements and interactions. In this work, we built a network of interconnected drugs, proteins and diseases and applied DrugNet to different types of tests for drug repositioning. RESULTS: We tested the performance of our approach on different validation tests, including cross validation and tests based on real clinical trials. DrugNet achieved a mean AUC value of 0.9552±0.0015 in 5-fold cross validation tests, and a mean AUC value of 0.8364 for tests based on recent clinical trials (phases 0-4) not present in our data. These results suggest that DrugNet could be very useful for discovering new drug uses. We also studied specific cases of particular interest, proving the benefits of heterogeneous data integration in this problem. CONCLUSIONS: Our methodology suggests that new drugs can be repositioned by generating ranked lists of drugs based on a given disease query or vice versa. Our study shows that the simultaneous integration of information about diseases, drugs and targets can lead to a significant improvement in drug repositioning tasks. DrugNet is available as a web tool from http://genome2.ugr.es/drugnet/ (accessed 23.09.14). Matlab source code is also available on the website.

CisMiner: genome-wide in-silico cis-regulatory module prediction by fuzzy itemset mining.

Eukaryotic gene control regions are known to be spread throughout non-coding DNA sequences which may appear distant from the gene promoter. Transcription factors are proteins that coordinately bind to these regions at transcription factor binding sites to regulate gene expression. Several tools allow to detect significant co-occurrences of closely located binding sites (cis-regulatory modules, CRMs). However, these tools present at least one of the following limitations: 1) scope limited to promoter or conserved regions of the genome; 2) do not allow to identify combinations involving more than two motifs; 3) require prior information about target motifs. In this work we present CisMiner, a novel methodology to detect putative CRMs by means of a fuzzy itemset mining approach able to operate at genome-wide scale. CisMiner allows to perform a blind search of CRMs without any prior information about target CRMs nor limitation in the number of motifs. CisMiner tackles the combinatorial complexity of genome-wide cis-regulatory module extraction using a natural representation of motif combinations as itemsets and applying the Top-Down Fuzzy Frequent- Pattern Tree algorithm to identify significant itemsets. Fuzzy technology allows CisMiner to better handle the imprecision and noise inherent to regulatory processes. Results obtained for a set of well-known binding sites in the S. cerevisiae genome show that our method yields highly reliable predictions. Furthermore, CisMiner was also applied to putative in-silico predicted transcription factor binding sites to identify significant combinations in S. cerevisiae and D. melanogaster, proving that our approach can be further applied genome-wide to more complex genomes. CisMiner is freely accesible at: http://genome2.ugr.es/cisminer. CisMiner can be queried for the results presented in this work and can also perform a customized cis-regulatory module prediction on a query set of transcription factor binding sites provided by the user.

Terapia secuencial con medicamentos. Conversión de la vía intravenosa a la vía oral. ¿Una buena estrategia para disminuir la bacteriemia relacionada con catéter? / Sequential drug therapy. Switching from the intravenous to the oral route. A good strategy for reducing catheter-related bacteremia?

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ProphNet: a generic prioritization method through propagation of information.

BACKGROUND: Prioritization methods have become an useful tool for mining large amounts of data to suggest promising hypotheses in early research stages. Particularly, network-based prioritization tools use a network representation for the interactions between different biological entities to identify novel indirect relationships. However, current network-based prioritization tools are strongly tailored to specific domains of interest (e.g. gene-disease prioritization) and they do not allow to consider networks with more than two types of entities (e.g. genes and diseases). Therefore, the direct application of these methods to accomplish new prioritization tasks is limited. RESULTS: This work presents ProphNet, a generic network-based prioritization tool that allows to integrate an arbitrary number of interrelated biological entities to accomplish any prioritization task. We tested the performance of ProphNet in comparison with leading network-based prioritization methods, namely rcNet and DomainRBF, for gene-disease and domain-disease prioritization, respectively. The results obtained by ProphNet show a significant improvement in terms of sensitivity and specificity for both tasks. We also applied ProphNet to disease-gene prioritization on Alzheimer, Diabetes Mellitus Type 2 and Breast Cancer to validate the results and identify putative candidate genes involved in these diseases. CONCLUSIONS: ProphNet works on top of any heterogeneous network by integrating information of different types of biological entities to rank entities of a specific type according to their degree of relationship with a query set of entities of another type. Our method works by propagating information across data networks and measuring the correlation between the propagated values for a query and a target sets of entities. ProphNet is available at: http://genome2.ugr.es/prophnet. A Matlab implementation of the algorithm is also available at the website.

Habilidades emocionales para el cuidado de enfermos esquizofrénicos / Emotional Abilities for Schizophrenic Patients´ Care Givers

Objetivo: determinar el nivel de desarrollo de las habilidades emocionales para el cuidado que poseen cuidadores principales de enfermos esquizofrénicos. Métodos: estudio no experimental, descriptivo. La muestra, no probabilística, se conformó con 31 cuidadores del San Cristóbal, Artemisa. Se recolectó información mediante el método empírico: cuestionario Habilidades Emocionales para el Cuidado del enfermo Esquizofrénico (HECE). Los datos fueron analizados mediante el método estadístico, fundamentalmente, el análisis porcentual. Resultados: existe un predominio de niveles de desarrollo insuficiente de las habilidades emocionales, expresándose en más de la mitad de los cuidadores. El indicador que mostró resultados más satisfactorios fue el reconocimiento de las propias emociones ante síntomas y comportamientos problemáticos del enfermo y las exigencias propias de la tarea de cuidar. El indicador más deficiente fue emitir respuestas ajustadas ante las emociones y sentimientos del enfermo esquizofrénico, con mayor porcentaje de insuficiencia en casi la mitad de la muestra. Los estados emocionales negativos fueron más representativos ante los síntomas y comportamientos problemáticos del enfermo, mientras que los positivos se asociaron al cumplimiento de las exigencias de la tarea de cuidar. Las principales estrategias de autocontrol emocional fueron la búsqueda de la distracción y la racionalización positiva. Los cuidadores pueden reconocer con frecuencia los estados emocionales de los enfermos, y la estrategia más empleada por los mismos para conseguir el control del enfermo resultó ser el apoyo emocional. Conclusiones: Estos resultados confirmaron la importancia de potenciar el desarrollo de habilidades emocionales en estos cuidadores(AU)

Objective: to determine the development of the emotional abilities that has the care givers of schizophrenic patients. Methods: a descriptive, non experimental study was done. The non probabilistic sample was formed by 31 care givers from San Cristobal, Artemisa. Through the empiric method, the questionnaire about emotional abilities for schizophrenic patients ‘care givers (HECE), the information was gathered. Data was analyzed through the statistical method, mainly through the porcentual analysis. Results: there is a predominance of insufficient developing levels of emotional abilities in more than the half of care givers. The indicator that showed better results was the recognition of the own emotions and problematic behaviors as well as the proper demands of the task of taking care. The more deficient indicator was the one related to issue adjust answers before the emotions and feelings of the schizophrenic patient with a major percentage of insufficiency in almost the half part of the sample. Negative emotions were the most representative symptoms considering the patient´s problematic behaviors and symptoms, meanwhile the positive symptoms were related to the achievement of the demands in the task of taking care. The main strategies of self control were distraction and positive appliance of logic. The care givers can frequently recognize the emotional states of the patients and the strategy more used to obtain the patient´s control was the same emotional support. Conclusions: these results confirmed the importance of promoting the development of emotional abilities on these care givers(AU)

Microarray profiling of mononuclear peripheral blood cells identifies novel candidate genes related to chemoradiation response in rectal cancer.

Preoperative chemoradiation significantly improves oncological outcome in locally advanced rectal cancer. However there is no effective method of predicting tumor response to chemoradiation in these patients. Peripheral blood mononuclear cells have emerged recently as pathology markers of cancer and other diseases, making possible their use as therapy predictors. Furthermore, the importance of the immune response in radiosensivity of solid organs led us to hypothesized that microarray gene expression profiling of peripheral blood mononuclear cells could identify patients with response to chemoradiation in rectal cancer. Thirty five 35 patients with locally advanced rectal cancer were recruited initially to perform the study. Peripheral blood samples were obtained before neaodjuvant treatment. RNA was extracted and purified to obtain cDNA and cRNA for hybridization of microarrays included in Human WG CodeLink bioarrays. Quantitative real time PCR was used to validate microarray experiment data. Results were correlated with pathological response, according to Mandard´s criteria and final UICC Stage (patients with tumor regression grade 1-2 and downstaging being defined as responders and patients with grade 3-5 and no downstaging as non-responders). Twenty seven out of 35 patients were finally included in the study. We performed a multiple t-test using Significance Analysis of Microarrays, to find those genes differing significantly in expression, between responders (n = 11) and non-responders (n = 16) to CRT. The differently expressed genes were: BC 035656.1, CIR, PRDM2, CAPG, FALZ, HLA-DPB2, NUPL2, and ZFP36. The measurement of FALZ (p = 0.029) gene expression level determined by qRT-PCR, showed statistically significant differences between the two groups. Gene expression profiling reveals novel genes in peripheral blood samples of mononuclear cells that could predict responders and non-responders to chemoradiation in patients with locally advanced rectal cancer. Moreover, our investigation added further evidence to the importance of mononuclear cells' mediated response in the neoadjuvant treatment of rectal cancer.

Evolution over a 15-year period of clinical characteristics and outcomes of critically ill patients with community-acquired bacteremia.

OBJECTIVE: In recent years, outcomes for critically ill patients with severe sepsis have improved; however, no data have been reported about the outcome of patients admitted for community-acquired bacteremia. We aimed to analyze the changes in the prevalence, characteristics, and outcome of critically ill patients with community-acquired bacteremia over the past 15 yrs. DESIGN: A secondary analysis of prospective cohort studies in critically ill patients in three annual periods (1993, 1998, and 2007). SETTING: Forty-seven ICUs at secondary and tertiary care hospitals. PATIENTS: All adults admitted to the participating ICUs with at least one true-positive blood culture finding within the first 48 hrs of admission. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 829 patients was diagnosed with community-acquired bacteremia during the study periods (148, 196, and 485 in the three periods). The prevalence density rate of community-acquired bacteremia increased from nine per 1000 ICU admissions in 1993 to 24.4 episodes per 1,000 ICU admissions in 2007 (p < 0.001). The prevalence of septic shock also increased from 4.6 episodes/1,000 admissions in 1993 to 14.6 episodes/1,000 admissions in 2007 (p < 0.001). Patients with community-acquired bacteremia were significantly older and had more comorbidities. No significant differences were observed in the presence of Gram-positive and Gram-negative micro-organisms among the three study periods. Mortality related to community-acquired bacteremia decreased over the three study periods: 42%, 32.2%, and 22.9% in 1993, 1998, and 2007, respectively (p < 0.01). The occurrence of septic shock and the number of comorbidities were independently associated with worse outcome. Appropriate antibiotic therapy and development of community-acquired bacteremia in 1998 and 2007 were independently associated with better survival. CONCLUSIONS: The prevalence of community-acquired bacteremia in ICU patients has increased. Despite a higher percentage of more severe and older patients, the mortality associated with community-acquired bacteremia decreased. Improved management of severe sepsis might explain the improvements in outcomes.

Value of ß-D-glucan and Candida albicans germ tube antibody for discriminating between Candida colonization and invasive candidiasis in patients with severe abdominal conditions.

PURPOSE: To assess the value of (1→3)-ß-D: -glucan (BDG), Candida albicans germ tube antibody (CAGTA), C-reactive protein (CRP), and procalcitonin (PCT) levels for the diagnosis of invasive candidiasis (IC) and for differentiating Candida spp. colonization from infection in ICU patients with severe abdominal conditions (SAC). METHODS: Prospective study of 176 non-neutropenic patients, with SAC at ICU admission, and expected to stay at least 7 days. Surveillance cultures and BDG, CAGTA, CRP, and PCT levels were performed on the third day of ICU stay and twice a week for four consecutive weeks. Patients were grouped into invasive candidiasis (IC), Candida colonization, and neither colonized/nor infected. The classification and regression tree (CART) analysis was used to predict IC in colonized patients. The discriminatory ability of the obtained prediction rule was assessed by the area under the ROC curve (AUC). RESULTS: The probabilities of IC were 59.3 % for the terminal node of BDG greater than 259 pg/mL and 30.8 % for BDG less than 259 pg/mL and CAGTA positivity, whereas there was a 93.9 % probability in predicting the absence of IC for BDG less than 259 pg/mL and negative CAGTA. Using a cutoff of 30 % for IC probability, the prediction rule showed 90.3 % sensitivity, 54.8 % specificity, 42.4 % positive predictive value, and 93.9 % negative predictive value with an AUC of 0.78 (95 % confidence interval 0.76-0.81). Significant differences in CRP (p = 0.411) and PCT (p = 0.179) among the studied groups were not found. CONCLUSIONS: BDG with a positive test for CAGTA accurately differentiated Candida colonization from IC in patients with SAC, whereas CRP and PCT did not.

Genome-wide differential genetic profiling characterizes colorectal cancers with genetic instability and specific routes to HLA class I loss and immune escape.

AIM: We compared the expression of genes related to inflammatory and cytotoxic functions between MSI and MSS (HLA-class I-negative and HLA-class I-positive) colorectal cancers (CRCs), seeking evidence of differences in inflammatory mediators and cytotoxic T-cell responses. Twenty-two CRCs were divided into three study groups as a function of HLA class I expression and MSI phenotype: 8 MSI tumours, 6 MSS/HLA- tumours and 6 MSS/HLA+ tumours (controls). FINDINGS: A first comparison between eight MSI and six MSS/HLA-positive (control) cancers, based on microarray analysis on an Affymetrix(®) HG-U133-Plus-PM plate, identified 1974 differentially expressed genes (P < 0.05). We grouped genes in Gene Ontology functional categories: apoptotic programme (72 genes, P = 5.5·10(-3)), leucocyte activation (43 genes, P = 1.8·10(-5)), T-cell activation (24 genes, P = 6.3·10(-4)), inflammatory response (40 genes, 2.3·10(-2)) and cytokine production (10 genes, P = 1.9·10(-2)). Real-time PCR and immunohistochemical evaluation were used to validate the data, finding that increased mRNA levels of pro-inflammatory cytokines and cytotoxic mediators were associated with greater infiltration by CD8+T lymphocytes in the MSI group (P < 0.001). Finally, HLA-class I-negative tumours were not grouped together but rather in accordance with features of the gene expression profile of MSI or MSS tumours. As expected, genes associated with antigen processing machinery and MHC class I molecules (TAP2, B2m) were downregulated in MSS/HLA-class I-negative CRCs (n = 6) in comparison to controls. CONCLUSIONS: In conclusion, microarray and immunohistochemical data may be useful to comprehensively assess tumour-host interactions and differentiate MSI from MSS cancers. The two types of tumour, MSI/HLA-class I-negative and MSS/HLA-class I-negative, showed marked differences in the composition and intensity of infiltrating leucocytes, suggesting that their immune escape strategies involve distinct pathways.

Latin American consensus: children born small for gestational age.

BACKGROUND: Children born small for gestational age (SGA) experience higher rates of morbidity and mortality than those born appropriate for gestational age. In Latin America, identification and optimal management of children born SGA is a critical issue. Leading experts in pediatric endocrinology throughout Latin America established working groups in order to discuss key challenges regarding the evaluation and management of children born SGA and ultimately develop a consensus statement. DISCUSSION: SGA is defined as a birth weight and/or birth length greater than 2 standard deviations (SD) below the population reference mean for gestational age. SGA refers to body size and implies length-weight reference data in a geographical population whose ethnicity is known and specific to this group. Ideally, each country/region within Latin America should establish its own standards and make relevant updates. SGA children should be evaluated with standardized measures by trained personnel every 3 months during year 1 and every 6 months during year 2. Those without catch-up growth within the first 6 months of life need further evaluation, as do children whose weight is ≤ -2 SD at age 2 years. Growth hormone treatment can begin in SGA children > 2 years with short stature (< -2.0 SD) and a growth velocity < 25th percentile for their age, and should continue until final height (a growth velocity below 2 cm/year or a bone age of > 14 years for girls and > 16 years for boys) is reached. Blood glucose, thyroid function, HbA1c, and insulin-like growth factor-1 (IGF-1) should be monitored once a year. Monitoring insulin changes from baseline and surrogates of insulin sensitivity is essential. Reduced fetal growth followed by excessive postnatal catch-up in height, and particularly in weight, should be closely monitored. In both sexes, gonadal function should be monitored especially during puberty. SUMMARY: Children born SGA should be carefully followed by a multidisciplinary group that includes perinatologists, pediatricians, nutritionists, and pediatric endocrinologists since 10% to 15% will continue to have weight and height deficiency through development and may benefit from growth hormone treatment. Standards/guidelines should be developed on a country/region basis throughout Latin America.

HER2 status in breast cancer: experience of a Spanish National Reference Centre.

BACKGROUND: Accurate HER2 testing is of great clinical value for the identification of breast cancer patients who are eligible for trastuzumab therapy. The aim of this study is to review breast carcinomas diagnosed from 2001 to 2007 at a Spanish National Reference Centre for HER2 testing, evaluating the agreement between HER2 immunohistochemical (IHC) tests and fluorescence in situ hybridisation (FISH) tests. METHODS: Demographic and clinical information was obtained from 2751 breast carcinoma patients. HER2 IHC and FISH tests were performed both in a local laboratory and in the reference centre. The HER2 IHC0/1+, IHC2+, IHC3+ and FISH-positive patients comprised 64%, 20%, 16% and 24% of the available population, respectively (results from the reference centre). Using statistical approaches, we evaluated the agreement between: (1) HER2 IHC and FISH tests, and (2) results provided by the local and the reference laboratories. RESULTS: The data confirmed a statistically significant relation between HER2 overexpression and amplification. We also found that instances of polysomy 17 and heterogeneous patterns of HER2 expression (heterogeneous staining distribution in different areas of the same tumour) are more frequently observed in HER2-positive tumours. Finally, since the diagnoses were made from 2001 to 2007, we could also observe a rising agreement rate between laboratories/pathologists with time. CONCLUSIONS: HER2 testing is most accurate when performed by experienced pathologists and at a high-volume reference laboratory. Polysomy 17 and HER2 heterogeneous staining patterns should also be considered for a better understanding of the variation in the anti-HER2 therapeutic response.

Health-care-associated bloodstream infections at admission to the ICU.

BACKGROUND: Infections occurring among outpatients having recent contact with the health-care system have been recently classified as health-care-associated infections to distinguish them from hospital- and community-acquired infections. Patients with bloodstream infections (BSIs) were studied to assess health-care-associated infections at admission in the ICU. METHODS: This work was a multicenter, prospective, observational study of all adult patients with BSI at ICU admission at 27 Spanish hospitals and one Argentine hospital. Cases of BSI were classified as community-acquired BSI (CAB), health-care-associated BSI (HCAB), or hospital-acquired BSI (HAB), and their characteristics were compared. RESULTS: Of 726 BSIs, 343 (47.2%) were CABs, 252 (34.7%) were HABs, and 131 (18.0%) were HCABs. Potentially antibiotic-resistant pathogens were more frequently isolated in HABs (34.8%) and HCABs (27.6%) than in CABs (10.3%) (P < .001). Logistic regression analysis revealed that HABs (OR, 4.6; 95% CI, 2.9-7.3), HCABs (OR, 3.1; 95% CI, 1.8-5.4), and BSIs of unknown origin (OR, 1.7; 95% CI, 1.0-2.8) were independently associated with the isolation of potentially antibiotic-resistant pathogens. The incidence of inappropriate treatment was significantly higher in HABs (OR, 3.4; 95% CI, 2.1-5.3) and in HCABs (OR, 1.8; 95% CI, 1.0-3.2) than in CABs. CONCLUSIONS: One in five BSIs diagnosed at ICU admission is health-care-associated. The incidence of potentially drug-resistant pathogens in HCABs is more similar to that of HABs, and they should be treated as such until culture data are available.

Genome-wide gene expression analysis in mouse embryonic stem cells.

Embryonic stem cell studies have generated great interest, due to their ability to form a wide variety of matured cells. However, there remains a poor understanding of mechanisms regulating the cell state of embryonic stem cells (ESCs) and of the genes they express during early differentiation. Gene expression analysis may be a valuable tool to elucidate either the molecular pathways involved in self-renewal and pluripotency, or early differentiation and to identify potential molecular therapy targets. The aim of this study was to characterize at the molecular level the undifferentiated mouse ESC state and the early development towards embryoid bodies. To attempt this issue, we performed CodeLink Mouse Uniset I 20K bioarrays in a well-characterized mouse ESC line, MES3, 3- and 7 day-old embryoid bodies and we compared our findings with those in adult tissue cells. Gene expression results were subsequently validated in a commercial stem cell line, CGR8 (ATCC). Significance Analysis of Microarrays (SAM) was used to identify statistically significant changes in microarray data. We identified 3664 genes expressed at significantly greater levels in MES3 stem cells than in adult tissue cells, which included 611 with 3-fold higher gene expression levels versus the adult cells. We also investigated the gene expression profile during early embryoid body formation, identifying 2040 and 2243 genes that were up-regulated in 3- and 7- day-old embryoid bodies, respectively. Our gene expression results in MES3 cells were partially confirmed in CGR8 cells, showing numerous genes that are expressed in both mouse stem cells. In conclusion, our results suggest that commonly expressed genes may be strong candidates for involvement in the maintenance of a pluripotent and undifferentiated phenotype and in early development.