[Glossopharyngeal neuralgia associated with syncopes and secondary to neck carcinoma]. / Neuralgia glosofaringea asociada a sincopes y secundaria a carcinoma de cuello.

TITLE: Neuralgia glosofaringea asociada a sincopes y secundaria a carcinoma de cuello.

An Update on Src Family of Nonreceptor Tyrosine Kinases Biology.

The members of the Src family of nonreceptor tyrosine kinases (SFKs) are implicated in multiple signaling processes that regulate key cellular functions, including proliferation, migration, differentiation, and survival. SFKs are activated by a large number of receptors for growth factors, cytokines, steroid hormones, G protein-coupled receptors, and also by adhesion proteins and other signaling partners. Through their common modular kinase an adapter protein domains, SFKs critically contribute to diversify different signal inputs, weaving a complex and dynamic network of cellular responses. Not surprisingly, SFKs are involved in embryo development and in the maintenance of different adult tissues and organs. Conversely, dysfunction of SFKs is associated with different pathologies, including cancer. Despite the continuous research in the field, several aspects of SFKs regulation and function are still not well defined and new roles for these proteins are steadily reported. The aim of this review is to provide an update on the major regulatory mechanisms of SFKs activity, including the emerging redox-dependent pathway. We have also focused on the functional implications of SFKs in Prolactin signaling and breast cancer development, two increasingly important aspects of SFKs biology. Finally, we briefly revisited the role of SFKs during embryo development and provide insights on the involvement of these proteins in the regulation of embryonic, somatic, and breast cancer stem cells.

Optimización de cultivos primarios de células de carcinoma renal de células claras como modelo "in vitro" para estudios metabólicos y determinantes de progresión neoplásica / Optimization of primary cultures of clear cell renal carcinoma cells as an "in vitro" model for metabolic studies and determinants of neoplastic progression

El objetivo del presente estudio fue optimizar la implementación de cultivos primarios a partir de muestras de carcinoma renal de células claras (CRCC) para comprobar la conservación del fenotipo lipogénico contra cortes fijados del mismo origen. Se utilizaron muestras de pacientes con CRCC, evaluándose diversas metodologías y condiciones experimentales de digestión de muestras, adherencia y despegue celular, fenotipo lipogénico, potencial de clonación, proliferación y capacidad de migración. El mayor rendimiento y viabilidad celular se verificó mediante digestión con colagenasa. La adherencia inicial se logró a las 24 hs de incubación, utilizando placas plásticas de cultivo, recubiertas con colágeno comercial y gelatina 0,2% en la mayoría de las muestras analizadas (60% de los casos). Se obtuvieron monocapas, con potencial de migración, en un 40% de los casos, tras 5 ± 1 días de incubación. El promedio de subcultivos fue de 3 ± 1. Este estudio permitió estandarizar cultivos primarios de CRCC comprobándose la conservación de la fenotipia lipogénica, logrando de dicha manera una herramienta importante y útil para el estudio de la biología tumoral y el ensayo de nuevas terapéuticas

The aim of this study was to optimize the implementation of primary cultures from samples of renal clear cell carcinoma (CRCC) to check the conservation of the lipogenic phenotype. CRCC Patient samples were used, in order to evaluate different methodologies and the experimental conditions of sample digestion, cell adhesion and lipogenic phenotype, proliferation and migration ability. The highest yield in cell number and viability was assessed using collagenase digestion. The initial adhesion was achieved after 24 hours of incubation in plastic plates recoverd with commercial collagen or 0.2% gelatin (60% of cases). Monolayers, with migration potential, were obtained in 40% of all cases, after 5 ± 1 days of incubation. The subcultures average was 3 ± 1. This study allowed us to standardize primary cultures of CRCC and check the conservation of the lipogenic phenotyping, achieving in this way an important and useful tool to study the tumor biology.

O objetivo deste trabalho foi otimizar a implementação de culturas primárias de amostras de carcinoma de células claras renal (CRCC) para verificar conservação fenótipo lipogenic contra os cortes previstos a mesma origem. As amostras dos pacientes foram utilizados CRCC, avaliando diferentes metodologias e as condições experimentais da digestão de amostras, adesão celular e fenótipo clonagem potencial take-lipogenic, proliferação e capacidade de migração. O maior rendimento e a viabilidade celular foi avaliada por digestão com colagenase. A adesão inicial foi obtida após 24 horas de incubação com colagénio e gelatina comercial 0,2% em 60% dos casos. As monocamadas foram obtidos em 40% após 5 ± 1 dias de incubação com o potencial de migração. As subculturas média foi de 3 ± 1. Este estudo nos permitiu padronizar culturas primárias de CRCC são verificados quanto à conservação da fenotipagem lipogenic, conseguindo desta forma um importante e útil para o estudo da biologia do tumor e teste de nova ferramenta terapêutica

Mediation analysis of an effective sexual health promotion intervention for Spanish adolescents.

The objective of this study is to determinate the factors that mediate in the self-reported consistent condom use over the 24-months post-intervention period in adolescents who received COMPAS, a sexual health promotion intervention targeted to Spanish adolescents. Twelve high schools located in Spain were randomized to an intervention or a control group with baseline, immediate-post, 12 and 24-month post-intervention assessments. Self-reported consistent condom use by 24 months post-intervention was the primary outcome. Based on the theory of planned behavior, we identified which theory-based variables mediated the intervention's effect on consistent condom use. Serial multiple mediation analysis indicated that attitudes toward condom use, when there are obstacles to use it, and self-efficacy mediated the COMPAS's effect in increasing consistent condom use. This is the first study that identifies the theoretical constructs that mediate the efficacy of a school-based intervention to promote sexual health in adolescents from Spain.

Nuevos modelos experimentales para el estudio de la homeostasis y la enfermedad cutánea / New Experimental Models of Skin Homeostasis and Diseases

La homeostasis de la piel, cuya regulación molecular es aún bastante desconocida, está íntimamente relacionada con la función de las células madre epidérmicas. El programa SkinModel-CM, auspiciado por la Comunidad de Madrid, reúne 5 grupos de investigación con el propósito de desarrollar nuevos modelos experimentales in vitro e in vivo para analizar la función de ADN metiltransferasa 1, la endoglina y la podoplanina en la actividad de las células madre epidérmicas y en la homeostasis y el cáncer cutáneos. Estos nuevos modelos comprenden tanto cultivos organotípicos 3 D, como ratones inmunodeficientes con la piel humanizada y ratones modificados genéticamente. Otro objetivo del programa es el uso de ratones con la piel humanizada como modelo para reconstruir enfermedades cutáneas, tales como el síndrome de Gorlin y el xeroderma pigmentoso, con el objeto de optimizar nuevos protocolos de intervención mediante la terapia fotodinámica (AU)

Homeostasis, whose regulation at the molecular level is still poorly understood, is intimately related to the functions of epidermal stem cells. Five research groups have been brought together to work on new in vitro and in vivo skin models through the SkinModel-CM program, under the auspices of the Spanish Autonomous Community of Madrid. This project aims to analyze the functions of DNA methyltransferase 1, endoglin, and podoplanin in epidermal stem cell activity, homeostasis, and skin cancer. These new models include 3-dimensional organotypic cultures, immunodeficient skin-humanized mice, and genetically modified mice. Another aim of the program is to use skin-humanized mice to model dermatoses such as Gorlin syndrome and xeroderma pigmentosum in order to optimize new protocols for photodynamic therapy (AU)

Cryptomphalus aspersa mollusc eggs extract promotes migration and prevents cutaneous ageing in keratinocytes and dermal fibroblasts in vitro.

BACKGROUND: The search of substances that minimize cutaneous ageing has increased in the last few years. Previous studies have described the regenerative properties of the secretion of the mollusc Cryptomphalus aspersa (C. aspersa) when applied topically. OBJECTIVE: We evaluate the in vitro effects of a new product derived from the eggs of C. aspersa, IFC-CAF, on cell proliferation, migration, distribution of cytoskeletal proteins, production of extracellular components as well as its ability to prevent cutaneous ageing because of intrinsic or extrinsic factors (exposure to UVB) by determination of ageing markers. METHODS: We have used the human keratinocyte cell line (HaCaT cells), primary dermal fibroblasts (HDF) and senescent dermal fibroblasts (SHDF). The effects of the compound on cell proliferation and on the cell cycle were determined by the MTT colorimetric assay, estimation of total protein and/or trypan blue test and by flow cytometry, respectively. We also studied cell migration using the wound-healing migration assay, whereas ELISA assays, Western Blot and immunofluorescence microscopy were carried out to test the expression of proteins related to cytoskeleton, extracellular matrix and with ageing. RESULTS: We have found that IFC-CAF does not promote proliferation but induces migration of HaCaT, HDF and SHDF in a time- and dose-dependent manner; a better organization of cytoskeletal proteins (F-actin and vimentin) and promotes the production of extracellular components (fibronectin, collagen 1 and MMPs) and the adhesion to cell-substrate vinculin protein. IFC-CAF also prevents cutaneous ageing. The treatment decreases the expression of the ageing-related markers b-Gal, p53 and p16INK4 in SDDF cells, and improves cell survival after UVB irradiation and nuclear repair in HaCaT cells. CONCLUSION: IFC-CAF has regenerative properties and protects against ageing factors being, therefore, a potential therapeutic agent for treating or preventing skin ageing.

New experimental models of skin homeostasis and diseases.

Homeostasis, whose regulation at the molecular level is still poorly understood, is intimately related to the functions of epidermal stem cells. Five research groups have been brought together to work on new in vitro and in vivo skin models through the SkinModel-CM program, under the auspices of the Spanish Autonomous Community of Madrid. This project aims to analyze the functions of DNA methyltransferase 1, endoglin, and podoplanin in epidermal stem cell activity, homeostasis, and skin cancer. These new models include 3-dimensional organotypic cultures, immunodeficient skin-humanized mice, and genetically modified mice. Another aim of the program is to use skin-humanized mice to model dermatoses such as Gorlin syndrome and xeroderma pigmentosum in order to optimize new protocols for photodynamic therapy.

Spanish version of the Sleep Self-Report (SSR): factorial structure and psychometric properties.

BACKGROUND: The Sleep Self-Report (SSR) is a questionnaire initially created for use with a sample from the USA to assess sleep patterns and problems in school-aged children. The objective of this study was to validate the SSR among a Spanish sample. METHODS: Participants were 1228 Spanish children from 8 to 12 years of age who completed the questionnaires at school anonymously. RESULTS: Internal consistency was good (ω = 0.85). Convergent validity with anxiety (r = 0.54) and perceived welfare (r = -0.53) measures, and divergent validity with a measure of academic performance and positive influence of peers (r = -0.22) were acceptable. Exploratory analysis suggested a factorial structure composed by four subscales: sleep quality, sleep anxiety, bedtime refusal and sleep routines. Confirmatory analysis indicated a good fit for the model (RMSEA = 0.04; GFI = 0.95; AGFI = 0.93; χ(2)/gl = 2.48). CONCLUSIONS: The SSR has demonstrated to have good psychometric properties in the Spanish-speaking sample of children. The factorial structure supported by exploratory and confirmatory analysis examines the most relevant areas of sleep in children. The satisfactory psychometric properties support the use of the Spanish version of the SSR by researchers and clinicians.

Novel group A rotavirus G8 P[1] as primary cause of an ovine diarrheic syndrome outbreak in weaned lambs.

Rotavirus is a worldwide major cause of diarrhea outbreaks in neonatal ruminants. An outbreak of ovine diarrheic syndrome (ODS) in 50-75 days-old lambs (weaned lambs) is described. Fecal immunochromatography and intestinal immunohistochemistry for rotavirus group A were performed. In addition, semi-nested multiplex RT-PCR for G and P rotavirus genotyping in combination with sequencing were performed, to support the diagnosis and identify the viral strain. A novel ovine rotavirus group A G8 P[1] strain was determined as the main cause of the ODS observed, whereas other pathogens were ruled out.

Wnt signalling and cancer stem cells

Intracellular signalling mediated by secreted Wnt proteins is essential for the establishment of cell fates and proper tissue patterning during embryo development and for the regulation of tissue homeostasis and stem cell function in adult tissues. Aberrant activation of Wnt signalling pathways has been directly linked to the genesis of different tumours. Here, the components and molecular mechanisms implicated in the transduction of Wnt signal, along with important results supporting a central role for this signalling pathway in stem cell function regulation and carcinogenesis will be briefl y reviewed (AU)

Hedgehog signalling as a target in cancer stem cells

Hedgehog (Hh) is one of the most important signalling pathways. Together with the Wnt, TGF-Beta/BMP and Notch pathways, it is involved in both embryonic development and adult tissue homeostasis. This is because Hh plays a central role in the proliferative control and differentiation of both embryonic stem cells and adult stem cells. In this way, an alteration in the Hh pathway, either by misexpression of components of that pathway or by changes in the expression of other cellular components that interfere with the Hh signalling system, may trigger the development of several types of cancer. This occurs because normal stem cells or their intermediaries toward differentiated mature cells are not part of the normal proliferative/ differentiation balance and begin to expand without control, triggering the generation of the so-called cancer stem cells. In this review, we will focus on the molecular aspects and the role of Hh signalling in normal tissues and in tumour development (AU)

[Experiences with the Eurotest in neuropsychological examinations. A screening test]. / Experiencia con el Eurotest en la exploración neuropsicológica. Un test de cribado.

INTRODUCTION: The increasing number of foreign patients being attended in our health centres makes it necessary to have tests that are compatible among different countries, especially in Europe. The objective of this study is to compare the Eurotest with other screening tests and to determine whether there are any differences between European (other than Spanish) and Spanish patients. PATIENTS AND METHODS: We included patients who were referred because of complaints that were compatible with cognitive impairment and they were asked to carry out a protocol of tests that included the global deterioration scale (GDS), Folstein's MMSE (Mini-Mental State Examination), the Clock Test (CT) and the Eurotest. Epidemiological data were collected. The sample used in the study included 45 patients, 21 males and 24 females, with a mean age of 69.76 years; 34 were born in Spain and 11 abroad. RESULTS: Diagnoses were 17 with mild cognitive impairment, 8 with Alzheimer's disease, 6 with age-associated impairment, 5 with vascular dementia, 5 with a psychiatric disorder, 1 with mixed dementia and 1 with dementia with Lewy bodies. The distribution according to the GDS was as follows: 10 with a GDS of 2, 18 with a GDS of 3, and 17 with a GDS of 4. All the patients with a GDS 2 completed the tests correctly, those with a GDS 3 scored below the cut-off point 5 for the Eurotest, 4 for the MMSE and 3 for CT, and in the GDS 4 they were 9, 7 and 6, respectively. Results did not vary between Spaniards and foreigners, the kappa index between MMSE and the Eurotest was 0.39 and the Pearson coefficient was 0.67. CONCLUSION: The Eurotest was slightly more useful in patients with a greater degree of impairment.

Experiencia con el Eurotest en la exploración neuropsicológica. Un test de cribado / Experiences with the eurotest in neuropsychological examinations. A screening test

Introducción. El incremento de pacientes extranjeros atendidos en nuestras consultas hace necesario disponer de tests compatibles entre múltiples países, especialmente del ámbito europeo. El objetivo del estudio es comparar el Eurotest con otros tests de cribado y comprobar si existen diferencias entre pacientes europeos (no españoles) y españoles. Pacientes y métodos. Incluimos pacientes remitidos por presentar quejas compatibles con deterioro cognitivo; se les realizó un protocolo de tests para su estudio, que incluyó la escala global de deterioro (GDS), el MMSE (Mini-Mental state examination) de Folstein, el test del reloj (TR) y el Eurotest. Recogimos datos epidemiológicos. Fueron 45 pacientes, 21 hombres y 24 mujeres, con una edad media de 69,76 años, 34 nacieron en España y 11 fuera. Resultados. Los diagnósticos fueron 17 con deterioro cognitivo leve, 8 con enfermedad de Alzheimer, 6 con deterioro asociado a la edad, 5 con demencia vascular, 5 con trastorno psiquiátrico, 1 con demencia mixta y 1 con demencia por cuerpos de Lewy. La distribución según GDS fueron: 10 con GDS de 2, 18 con GDS de 3, y 17 con GDS de 4. Todos los pacientes con GDS 2 realizaron los tests correctamente, con GDS 3 dieron por debajo del punto de corte 5 para Eurotest, 4 para MMSE y 3 para TR, y en GDS 4 fueron 9, 7 y 6, respectivamente. Los resultados no variaron entre españoles y extranjeros, el índice kappa entre MMSE y Eurotest fue de 0,39, y el coeficiente de Pearson, de 0,67. Conclusión. El Eurotest fue ligeramente más útil en pacientes más deteriorados (AU)

Introduction. The increasing number of foreign patients being attended in our health centres makes it necessary to have tests that are compatible among different countries, especially in Europe. The objective of this study is to compare the Eurotest with other screening tests and to determine whether there are any differences between European (other than Spanish) and Spanish patients. Patients and methods. We included patients who were referred because of complaints that were compatible with cognitive impairment and they were asked to carry out a protocol of tests that included the global deterioration scale (GDS), Folstein’s MMSE (Mini-Mental State Examination), the Clock Test (CT) and the Eurotest. Epidemiological data were collected. The sample used in the study included 45 patients, 21 males and 24 females, with a mean age of 69.76 years; 34 were born in Spain and 11 abroad. Results. Diagnoses were 17 with mild cognitive impairment, 8 with Alzheimer's disease, 6 with age-associated impairment, 5 with vascular dementia, 5 with a psychiatric disorder, 1 with mixed dementia and 1 with dementia with Lewy bodies. The distribution according to the GDS was as follows: 10 with a GDS of 2, 18 with a GDS of 3, and 17 with a GDS of 4. All the patients with a GDS 2 completed the tests correctly, those with a GDS 3 scored below the cutoff point 5 for the Eurotest, 4 for the MMSE and 3 for CT, and in the GDS 4 they were 9, 7 and 6, respectively. Results did not vary between Spaniards and foreigners, the kappa index between MMSE and the Eurotest was 0.39 and the Pearson coefficient was 0.67. Conclusion. The Eurotest was slightly more useful in patients with a greater degree of impairmen (AU)

Epigenetic control of nuclear architecture.

The cell nucleus is a highly structured compartment where nuclear components are thought to localize in non-random positions. Correct positioning of large chromatin domains may have a direct impact on the localization of other nuclear components, and can therefore influence the global functionality of the nuclear compartment. DNA methylation of cytosine residues in CpG dinucleotides is a prominent epigenetic modification of the chromatin fiber. DNA methylation, in conjunction with the biochemical modification pattern of histone tails, is known to lock chromatin in a close and transcriptionally inactive conformation. The relationship between DNA methylation and large-scale organization of nuclear architecture, however, is poorly understood. Here we briefly summarize present concepts of nuclear architecture and current data supporting a link between DNA methylation and the maintenance of large-scale nuclear organization.

Epigenetic inactivation of the Wnt antagonist DICKKOPF-1 (DKK-1) gene in human colorectal cancer.

Colorectal cancer is a major cause of cancer death worldwide. A number of key oncogenes and tumor suppressor genes have been proposed to drive progression from healthy colonic epithelia to malignant tumors, including members of the Wnt/beta-catenin pathway. Recently, CpG island promoter hypermethylation was shown to cause inactivation of two extracellular Wnt inhibitors in colon cancer: secreted frizzled-related proteins (sFRPs) and Wnt inhibitory factor-1 (WIF-1). Here, we show for the first time that another extracellular Wnt inhibitor, the DICKKOPF-1 (DKK-1) gene, is transcriptionally silenced by CpG island promoter hypermethylation in colon cancer cell lines (n=9), whereas treatment with the DNA-demethylating agent 5-aza-2-deoxycytidine restored DKK-1 expression. Restoration of DKK-1 function in non-expressing cells bearing a truncated APC (Adenomatous Polyposis Coli) gene had no effect on beta-catenin/T-cell factor-dependent transcription, but induced tumor suppressor-like features such as reduced colony formation density and tumor growth inhibition in nude mice. These results suggest additional functions for DKK-1 other than inhibiting canonical Wnt signaling. In primary colorectal tumors, DKK-1 was found hypermethylated in 17% (nine of 54) of cases. Furthermore, while for both SFRP-1 and WIF-1 methylation-associated silencing occurred across the whole spectrum of colorectal tumorigenesis, DKK-1 promoter was selectively hypermethylated in advanced colorectal neoplasms (Duke's C and D tumors).