Evaluation of immunologic parameters in canine glioma patients treated with an oncolytic herpes virus.

Aim: To molecularly characterize the tumor microenvironment and evaluate immunologic parameters in canine glioma patients before and after treatment with oncolytic human IL-12-expressing herpes simplex virus (M032) and in treatment naïve canine gliomas. Methods: We assessed pet dogs with sporadically occurring gliomas enrolled in Stage 1 of a veterinary clinical trial that was designed to establish the safety of intratumoral oncoviral therapy with M032, a genetically modified oncolytic herpes simplex virus. Specimens from dogs in the trial and dogs not enrolled in the trial were evaluated with immunohistochemistry, NanoString, Luminex cytokine profiling, and multi-parameter flow cytometry. Results: Treatment-naive canine glioma microenvironment had enrichment of Iba1 positive macrophages and minimal numbers of T and B cells, consistent with previous studies identifying these tumors as immunologically "cold". NanoString mRNA profiling revealed enrichment for tumor intrinsic pathways consistent with suppression of tumor-specific immunity and support of tumor progression. Oncolytic viral treatment induced an intratumoral mRNA transcription signature of tumor-specific immune responses in 83% (5/6) of canine glioma patients. Changes included mRNA signatures corresponding with interferon signaling, lymphoid and myeloid cell activation, recruitment, and T and B cell immunity. Multiplexed protein analysis identified a subset of oligodendroglioma subjects with increased concentrations of IL-2, IL-7, IL-6, IL-10, IL-15, TNFα, GM-CSF between 14 and 28 days after treatment, with evidence of CD4+ T cell activation and modulation of IL-4 and IFNγ production in CD4+ and CD8+ T cells isolated from peripheral blood. Conclusion: These findings indicate that M032 modulates the tumor-immune microenvironment in the canine glioma model.

¿Repercutirán las nuevas cifras de las guías AHA/ACC en la definición y manejo de la hipertensión en Latinoamérica? / Will the new figures from the AHA/ACC guidelines on the definition and treatment of hypertension in Latin America have an impact?

Las nuevas guías de la Asociación Americana del Corazón (AHA) y del Colegio Americano del Corazón (ACC) han generado una importante discusión sobre los criterios diagnósticos y el manejo de hipertensión arterial (HTA), ya que estas nuevas guías proponen cambios importantes en la definición de la HTA, pasando su diagnóstico de cifras de presión arterial (PA) iguales o superiores a 140/90mmHg a cifras iguales o superiores a 130/80mmHg. Además, las nuevas guías proponen que las metas a alcanzar para definir el control adecuado de la HTA también sean más bajas, con cifras de PA menores de 130/80mmHg, con lo cual en términos globales se espera un incremento importante en el número de individuos considerados hipertensos, algunos de los cuales necesitarán más medicamentos para alcanzar las nuevas metas de adecuado control, todo lo que ha llevado al cuestionamiento sobre la viabilidad de la aplicabilidad clínica de estas nuevas guías, dado el enorme incremento financiero que significa el tratar con medicamentos a los nuevos millones de pacientes hipertensos. Además de estos inconvenientes prácticos, también se ha cuestionado la validez académica de las nuevas guías AHA/ACC, dado el hecho de que las recomendaciones emergen básicamente de los resultados obtenidos de un solo estudio, el cual tiene importantes diferencias metodológicas con la mayoría de estudios realizados previamente; además, sus resultados son inconsistentes con lo demostrado en otros estudios, lo que ha determinado que algunas sociedades científicas, como la Sociedad Latinoamericana de Hipertensión (LASH), mantengan las recomendaciones anteriores

Given the fact that new guidelines from the American Heart Association (AHA) and American College of Cardiology (ACC) propose important changes in the definition of hypertension, from equal or greater than 140/90mmHg to equal or greater than 130/80mmHg, major debate has been generated about the diagnostic criteria and the treatment of high blood pressure (HBP). In addition, these guidelines recommend that in order to achieve control of hypertension the goal should be a reading lower than 130/80mmHg. These new figures will significantly increase the amount of individuals considered hypertensive, some of whom will need more medication to achieve the new goal. This paradigm has led to questioning the goal's clinical applicability given the enormous financial burden that would result from treating millions of new hypertensive patients. The academic validity of the AHA-ACC guidelines has also been questioned given the fact that the recommendations emerged basically from results obtained from a single study with important methodological differences compared to the majority of studies carried out previously. Furthermore, its outcomes are inconsistent with those of previous studies. This conflict has led to some scientific institutions, such as the Latin American Society of Hypertension (LASH), continuing to adhere to the previous recommendations

[Will the new figures from the AHA/ACC guidelines on the definition and treatment of hypertension in Latin America have an impact?] / ¿Repercutirán las nuevas cifras de las guías AHA/ACC en la definición y manejo de la hipertensión en Latinoamérica?

Given the fact that new guidelines from the American Heart Association (AHA) and American College of Cardiology (ACC) propose important changes in the definition of hypertension, from equal or greater than 140/90mmHg to equal or greater than 130/80mmHg, major debate has been generated about the diagnostic criteria and the treatment of high blood pressure (HBP). In addition, these guidelines recommend that in order to achieve control of hypertension the goal should be a reading lower than 130/80mmHg. These new figures will significantly increase the amount of individuals considered hypertensive, some of whom will need more medication to achieve the new goal. This paradigm has led to questioning the goal's clinical applicability given the enormous financial burden that would result from treating millions of new hypertensive patients. The academic validity of the AHA-ACC guidelines has also been questioned given the fact that the recommendations emerged basically from results obtained from a single study with important methodological differences compared to the majority of studies carried out previously. Furthermore, its outcomes are inconsistent with those of previous studies. This conflict has led to some scientific institutions, such as the Latin American Society of Hypertension (LASH), continuing to adhere to the previous recommendations.

Community-wide measles outbreak in the Region of Madrid, Spain, 10 years after the implementation of the Elimination Plan, 2011-2012.

We describe a community-wide outbreak of measles due to a D4 genotype virus that took place in the Region of Madrid, Spain, between February 2011 and August 2012, along with the control measures adopted. The following variables were collected: date of birth, sex, symptoms, complications, hospital admission, laboratory test results, link with another cases, home address, places of work or study, travel during the incubation period, ethnic group, and Mumps-Measles-Rubella (MMR) vaccination status. Incidences were calculated by 100,000 inhabitants. A total of 789 cases were identified. Of all cases, 36.0% belonged to Roma community, among which 68.7% were 16 months to 19 y old. Non-Roma cases were predominantly patients from 6 to 15 months (28.1%) and 20 to 39 y (52.3%). Most cases were unvaccinated. We found out that 3.0% of cases were healthcare workers. The first vaccination dose was brought forward to 12 months, active recruitment of unvaccinated children from 12 months to 4 y of age was performed and the vaccination of healthcare workers and of members of the Roma community was reinforced. High vaccination coverage must be reached with 2 doses of MMR vaccine, aimed at specific groups, such as young adults, Roma population and healthcare workers.

Seroprevalence of measles and rubella virus antibodies in the population of the Community of Madrid, 2008-2009.

The seroprevalence (SP) of measles and rubella virus antibodies is presented by age groups obtained in the IV Serosurvey of the Region of Madrid (2008-2009). The target population is composed of residents with ages ranging between 2 and 60 years in the Region of Madrid. A two-stage cluster sample is used. The SP of measles virus antibodies is 97.8% (CI 95%: 97.3-98.2). The highest SP is observed in the 2-5 year and 41-60 year age groups. The point estimate does not reach 95% in the 16-20 and 21-30 year age groups. The SP of rubella virus antibodies is 97.2% (CI 95%: 96.5-97.7). The SP is over 95% in all of the age groups. In immigrant women between the ages of 16 and 49, the SP is 95.9% (CI 95%: 93.7-97.4). The identification of groups susceptible to the measles virus in young adults could lead to outbreaks as a result of importing the virus. The circulation of the rubella virus is possible among immigrant women aged between 16 and 49 years, which could lead to the appearance of SRC cases. Epidemiological surveillance will allow the impact on the measles and rubella elimination plan to be determined in the future.

Methamphetamine induces low levels of neurogenesis in striatal neuron subpopulations and differential motor performance.

Methamphetamine (METH) causes significant loss of some striatal projection and interneurons. Recently, our group reported on the proliferation of new cells 36 h after METH and some of the new cells survive up to 12 weeks (Tulloch et al., Neuroscience 193:162-169, 2011b). We hypothesized that some of these cells will differentiate and express striatal neuronal phenotypes. To test this hypothesis, mice were injected with METH (30 mg/kg) followed by a single BrdU injection (100 mg/kg) 36 h after METH. One week after METH, a population of BrdU-positive cells expressed the neuronal progenitor markers nestin (18 %) and ß-III-tubulin (30 %). At 8 weeks, 14 % of the BrdU-positive cells were also positive for the mature neuron marker, NeuN. At 12 weeks, approximately 7 % of the BrdU-positive cells co-labeled with ChAT, PV or DARPP-32. We measured motor coordination on the rotarod and psychomotor activity in the open-field. At 12 weeks, METH-injected mice exhibited delayed motor coordination deficits. In contrast, open-field tests revealed that METH-injected mice compared to saline mice displayed psychomotor deficits at 2.5 days but not at 2 or more weeks after METH. Taken together, these data demonstrate that some of the new cells generated in the striatum differentiate and express the phenotypes of striatal neurons. However, the proportion of these new neurons is low compared to the proportion that died by apoptosis 24 h after the METH injection. More studies are needed to determine if the new neurons are functional.

Novel heuristic search for ventricular arrhythmia detection using normalized cut clustering.

Processing of the long-term ECG Holter recordings for accurate arrhythmia detection is a problem that has been addressed in several approaches. However, there is not an outright method for heartbeat classification able to handle problems such as the large amount of data and highly unbalanced classes. This work introduces a heuristic-search-based clustering to discriminate among ventricular cardiac arrhythmias in Holter recordings. The proposed method is posed under the normalized cut criterion, which iteratively seeks for the nodes to be grouped into the same cluster. Searching procedure is carried out in accordance to the introduced maximum similarity value. Since our approach is unsupervised, a procedure for setting the initial algorithm parameters is proposed by fixing the initial nodes using a kernel density estimator. Results are obtained from MIT/BIH arrhythmia database providing heartbeat labelling. As a result, proposed heuristic-search-based clustering shows an adequate performance, even in the presence of strong unbalanced classes.

Unsupervised feature relevance analysis applied to improve ECG heartbeat clustering.

The computer-assisted analysis of biomedical records has become an essential tool in clinical settings. However, current devices provide a growing amount of data that often exceeds the processing capacity of normal computers. As this amount of information rises, new demands for more efficient data extracting methods appear. This paper addresses the task of data mining in physiological records using a feature selection scheme. An unsupervised method based on relevance analysis is described. This scheme uses a least-squares optimization of the input feature matrix in a single iteration. The output of the algorithm is a feature weighting vector. The performance of the method was assessed using a heartbeat clustering test on real ECG records. The quantitative cluster validity measures yielded a correctly classified heartbeat rate of 98.69% (specificity), 85.88% (sensitivity) and 95.04% (general clustering performance), which is even higher than the performance achieved by other similar ECG clustering studies. The number of features was reduced on average from 100 to 18, and the temporal cost was a 43% lower than in previous ECG clustering schemes.

Effect of latency on clustering of P300 recordings for ADHD discrimination.

This paper is focused on testing the latency contribution as regards the quality of formed groups for discriminating between healthy and attention deficit hyperactivity disorder children. To this end, two different cases are considered: nonaligned original recordings and aligned signals according to P300 position. For latter case, a novel approach to conduct time location of P300 component is introduced, which is based on derivative of event-related potential signals. The used database holds event-related potentials registered in auditory and visual oddball paradigm. Several experiments are carried out testing both configurations of considered data matrix. For grouping input data matrices, the k-means clustering technique is employed. To assess the quality of formed clusters and the relevance for clustering of latency-based features, relative values of distances between centroids and data points are computed in order to apprise separability and compactness of estimated clusters. Experimental results show that time localization of P300 component is not a decisive feature in formation of compact and well-defined groups within a discrimination framework for two considered data classes under certain conditions.

Molecular characterisation of KIR2DS2*005, a fusion gene associated with a shortened KIR haplotype.

KIR2DS2 is an activating homologue of KIR2DL2, an inhibitory killer-cell immunoglobulin-like receptor (KIR) that surveys expression of major histocompatibility complex-C allotypes bearing a C1 epitope. We have studied here its allele KIR2DS2*005, which shows a hybrid structure-it is identical to other KIR2DS2 alleles in the ectodomain, but has transmembrane and cytoplasmic regions identical to those of KIR2DS3(*)001, a short-tailed KIR of uncertain expression and function. Our results reveal that KIR2DS2*005 is a fusion gene-the product of an unequal crossing over by which the genes KIR2DS2 and KIR2DS3 recombined within a 400 base pair region of complete identity in intron 6. Also resulting from that recombination was a shortened KIR haplotype of the B group, in which three genes commonly linked to KIR2DS2 (KIR2DL2, KIR2DL5B and KIR2DS3) are deleted. Population studies indicate that KIR2DS2*005 is still associated to such haplotype, and it can be found in approximately 1.2% of Caucasoids. Using a combination of two monoclonal antibodies, we also demonstrate that KIR2DS2*005 encodes a molecule expressed on the surface of natural killer- and T-lymphocytes.

Weighted-PCA for unsupervised classification of cardiac arrhythmias.

A method that improves the feature selection stage for non-supervised analysis of Holter ECG signals is presented. The method corresponds to WPCA approach developed mainly in two stages. First, the weighting of the feature set through a weight vector based on M-inner product as distance measure and a quadratic optimization function. The second one is the linear projection of weighted data using principal components. In the clustering stage, some procedures are considered: estimation of the number of groups, initialization of centroids and grouping by means a soft clustering algorithm. In order to decrease the procedure computational cost, segment analysis, grouping contiguous segments and establishing union and exclusion criteria per each cluster, is carried out. This work is focused to classify cardiac arrhythmias into 5 groups, according to the standard of the AAMI (ANSI/AAMI EC57:1998/ 2003). To validate the method, some recordings from MIT/BIH arrhythmia database are used. By employing the labels of each recording, the performance is assessed with supervised measures (Se = 90.1%, Sp = 98.9% y Cp = 97.4%), enhancing other works in the literature that do not take into account all heartbeat types.

Unsupervised feature selection in cardiac arrhythmias analysis.

The problem of detecting clinical events related to cardiac arrhythmias in long term electrocardiograms is a difficult one due to the large amount of irrelevant information that hides such events. This problem has been addressed in the literature by means of clustering or classification algorithms that create data partitions according to a cost function based on heartbeat features dissimilarity measures. However, studies about the type or number of heartbeat features is lacking. Usually, the feature sets used are relevant but redundant, which degrades algorithm performance. This paper describes a method for automatic selection of heartbeat features. This method is assessed using real signals from the MIT database and common features used in previous works.

Multiple sclerosis associates with LILRA3 deletion in Spanish patients.

The genetic susceptibility to multiple sclerosis (MS) is only partially explained, and it shows geographic variations. We analyse here two series of Spanish patients and healthy controls and show that relapsing MS (R-MS) is associated with a gene deletion affecting the hypothetically soluble leukocyte immunoglobulin (Ig)-like receptor A3 (LILRA3, 19q13.4), in agreement with an earlier finding in German patients. Our study points to a gene-dose-dependent, protective role for LILRA3, the deletion of which synergizes with HLA-DRB1(*)1501 to increase the risk of R-MS. We also investigated whether the risk of suffering R-MS might be influenced by the genotypic diversity of killer-cell Ig-like receptors (KIRs), located only approximately 400 kb telomeric to LILRA3, and implicated in autoimmunity and defence against viruses. The relationship of LILRA3 deletion with R-MS is not secondary to linkage disequilibrium with a KIR gene, but we cannot exclude some contributions of KIR to the genetic susceptibility to R-MS.

The 5' intergenic, promoter, pseudoexon 3 and complete coding sequences of the hybrid gene KIR2DS3*002.

Genomic and mRNA sequences support the KIR2DS3*002 gene being a hybrid of KIR2DS3*00103 and KIR2DS5.

Duplication, mutation and recombination of the human orphan gene KIR2DS3 contribute to the diversity of KIR haplotypes.

The KIR2DS3 gene is an activating homologue of the inhibitory killer-cell immunoglobulin-like receptors (KIR) that recognize HLA-C molecules, enabling NK cells to survey the normal function of endogenous antigen presentation. The genetics of KIR2DS3 is complicated by the existence of alleles with similar coding sequences that map to different regions of the KIR complex in chromosome 19, or whose location in this complex is unknown. Here, by studying the family segregation of the KIR alleles 2DS3*001, *002 and *003N, and the distribution of these in unrelated individuals, we demonstrate the existence of two paralogous KIR2DS3 genes that can be inherited separately or, as it happens frequently in Caucasoids due to linkage disequilibrium, together. Each KIR2DS3 gene is almost invariably associated in its 5' end to a different copy of KIR2DL5, a gene previously shown to be duplicated in humans. KIR2DL5 and KIR2DS3 thus form two highly homologous gene clusters situated in the centromeric and the telomeric intervals of KIR haplotypes. Recombination between those clusters is the likely origin of new haplotypes, characterized in this study, which harbour further duplications or deletions of multiple KIR genes. Our results help understand the genetics of KIR2DS3 and the diversity of human KIR genotypes.

Crowding effect on adult growth, pre-patent period and egg shedding of Fasciola hepatica.

Fascioliasis pathogenesis depends on fluke burden. In human hyperendemic zones, individual infection intensities reach very high levels and the majority of infected subjects should be in the advanced chronic phase. The rat model offers a useful approach for pathological research in the advanced chronic period. The influence of infection intensity per rat on fluke development, pre-patent period and egg shedding (eggs/g faeces/worm) was analysed in 3 groups (I: 1-3 worms/rat; II: 4-6; III: 7-9). Ontogenetic trajectories of fluke body measures followed a logistic model. Results showed that when the burden increases, the maximum values of fluke measures decrease. The crowding effect is manifest when fluke measures approximate their maximums in the advanced chronic stage. The pre-patent period and egg production decrease when the burden increases. This means that measurements of eggs per gramme of faeces tend to underestimate the fluke burden. The present study demonstrates how to quantify the fascioliasis experimental rat model crowding effect on adult growth, pre-patent period and egg production. This quantification may be of great interest in epidemiological studies and in experimental research on the in vivo actions of different anthelminthic drugs and vaccines, pathology, immunology and resistance studies.

Hermafroditismo verdadero asociado a síndrome de insensibilidad a los andrógenos. Revisión de la literatura y diagnóstico diferencial / True hermaphroditism associated to androgenics insensitivity syndrome. Literature revision and diferential diagnosis

El hermafroditismo verdadero es una causa muy poco común de intersexualidad caracterizada por la coexistencia de tejido ovárico y testicular en el mismo individuo, independientemente de su cariotipo, manifestándose en la mayoría de las ocasiones como genitales ambiguos en el recién nacido. Es fundamental establecer un diagnóstico diferencial precoz con otros cuadros clínicos como la disgenesia gonadal mixta o el seudohermafroditismo masculino, para la correcta orientación terapéutica, así como para la asignación del género más apropiado lo antes posible, con objeto de facilitar el adecuado desarrollo físico, psicológico, social y sexual de la persona (AU)

Primary breast lymphoma: report of 2 cases / Primary breast lymphoma: report of 2 cases

El linfoma no hodgkiniano primario de mama (LNHPM) constituye un hallazgo muy poco común, representa un 0,04-0,53 por ciento de todos los tumores malignos de mama y aproximadamente un 2,2 por ciento de los linfomas extranodales. El diagnóstico se basa en los hallazgos clínicos y anatomopatológicos y exige un exhaustivo estudio de extensión. Las técnicas de imagen no han demostrado ser útiles en el diagnóstico precoz debido al rápido crecimiento del LNHPM y su similitud radiológica con otras tumoraciones mamarias. El fenotipo histológico es un factor determinante en la elección del tratamiento, esencialmente en los casos que precisa quimioterapia, que variarán dependiendo de cada entidad histopatológica. La radioterapia puede ser una opción como tratamiento complementario en la mama. La estadificación de Ann Arbor representa el único factor pronóstico estadísticamente significativo. A continuación, presentamos 2 casos de esta variedad clínica, diagnosticados en nuestro servicio en los 2 últimos años. El primer caso corresponde a un linfoma de Burkitt estadio IV A de Ann Arbor y el segundo a un linfoma difuso de células grandes B estadio I. En ambos casos se ha conseguido la remisión completa mediante tratamiento con exéresis local, radioterapia y quimioterapia, y no se ha evidenciado recurrencia ni progresión clínica tras un año de seguimiento. Hemos realizado una amplia revisión de la bibliografía al respecto, haciendo énfasis sobre su verdadera incidencia, factores pronósticos y combinaciones terapéuticas recomendadas. El LNHPM constituye un hallazgo muy poco frecuente, pero a pesar de su excepcionalidad, no debemos olvidar incluirlo en el diagnóstico diferencial de las tumoraciones mamaria (AU)

Cytotoxic effects of two antimolting insecticides in mammalian CHO-K1 cells.

Cytotoxicity of two insect growth regulators, diflubenzuron, a benzoylphenylurea derivative that inhibits the synthesis of new chitin in target organisms, and pyriproxyfen, an insect juvenile hormone analogue, were tested on CHO-K1 cultures, using the neutral red incorporation assay. Both compounds displayed cytotoxic effects that rise with time exposure. The presence of either fetal calf serum or bovine serum albumin diminished significantly the cytotoxicity of both compounds, thus pointing to a strong protein binding. In addition, extensive metabolization with rat liver submitochondrial fraction gave rise to metabolites less toxic than the parent compounds, implying the relative safety of both diflubenzuron and pyriproxyfen in mammals.