Rol de las metaloproteasas y caspasas en la conducta biológica del carcinoma basocelular. Role of the metalloproteinases and caspasa Biology in the conduct of basal cell carcinoma.

El carcinoma basocelular (CBC) representa el tumor maligno más frecuente de la piel en personas de raza blanca. Incide en mayores de 50 años de edad e infiltra los tejidos, pero rara vez ocasiona metástasis. En trabajos anteriores, observamos imágenes de cuerpos apoptóticos con MOAR (microscopía óptica de alta resolución), en pieles portadoras de CBC. Todas las células eucariotas poseen una maquinaria enzimática que interviene en la apoptosis. Estas enzimas son las proteincinasas pertenecientes a la familia de las caspasas. Se sintetizan como procaspasas, las cuales una vez activadas, actúan sobre otra caspasa en una reacción secuencial en cadena. Se reconoce que la apoptosis juega un rol importante en el balance y mantenimiento de los tejidos. Hipótesis: -Investigar si la expresión de metaloproteasas y caspasas en las células del CBC promueven y contribuyen a la progresión y extensión tumoral. Objetivos:-Profundizar el conocimiento de la conducta biológica del CBC, en referencia a la relación de los patrones de crecimiento macro y microscópicos del tumor, enfatizando en la expresión de metaloproteasas (colagenasa-3 y streptomelisina), CD-31, Ki-67, oncoproteína bcl-2 y caspasas, mediante la implementación de técnicas de inmunohistoquímica. -Analizar a nivel morfológico la presencia de cuerpos apoptóticos por medio de MOAR y determinar si influyen en la conducta biológica del tumor. Materiales y Métodos: Se realizará un estudio retrospectivo y prospectivo de pacientes asistidos en el Servicio de Dermatología del Hospital Nacional de Clínicas, desde el año 2000. Se utilizarán tomas incisionales de piel, que se fijarán en formol neutro al 10 por ciento y luego serán procesadas e incluidas en parafina y coloreadas con H-E (hematoxilina-eosina). Especimenes seleccionados, se utilizarán para técnicas de MOAR e inmunocitoquímica. El material para MOAR, se fijará en Karnovsky enfriado a pH 7,2 y se incluirá en resinas epóxicas, seccionados con un ultramicrótomo Porter BluMT1 (1micra) y coloreados con P.A.S. azul de toluidina, fuscina básica y metenamina-plata. La aplicación de las técnicas inmunohistoquímicas, se utilizaran sobre cortes desparafinados con xilol, que permitirán investigar en forma retrospectiva y prospectiva la matriz extracelular (MEC) y los elementos neoplásicos. Los anticuerpos monoclonales a utilizar serán: Colageno tipo IV clone CIV 22 M0785 monoclonal antibody (DAKO), CD44 clone E29 M0613 monoclonal antibody (DAKO), Oncoproteina bcl-2 clone 124 M0887 monoclonal antibody (DAKO), CD31: clone 35 BH11 MO631 monoclonal antibody (DAKO) K-167 clone PC10 M0879 monoclonal antibody.(DAKO) bclx y Caspasa 8. Metodología Estadística Los resultados serán evaluados mediante: a) Varianza Anova; para datos con distribución gausiana como media, SD. b) Krusal Walles; para rasgos de distribución no gausiana. c) Chi2 para determinar la diferencia significativa para proporciones. Resultados esperados: Del análisis histomorfológico cualitativo y cuantitativo de biopsias de piel con CBC, aportar determinaciones que clarifiquen la conducta biológica de este tumor.- Importancia del Proyecto: El CBC es una entidad que está aumentando su incidencia en forma exponencial, debido sobre todo a pautas sociales. No sólo ocasiona trastornos en los pacientes, sino que también, representa gastos en el sistema de salud. Asimismo, significa un desafío para la investigación. Esto permite explorar aspectos de la biología molecular, a partir de una entidad que posee características únicas.

PARTICIPACIÓN DEL RECONOCIMIENTO INMUNE INNATO EN LA RESPUESTA DE CARDIOMIOCITOS DURANTE LA INFECCIÓN EXPERIMENTAL CON TRYPANOSOMA CRUZI. Characterization of the cardiac innate immune response during Trypanosoma cruzi experimental infection.

La enfermedad de Chagas, causada por Trypanosoma cruzi, constituye la principal miocarditis infecciosa a nivel mundial. Crecientes evidencias revelan que la respuesta inmune innata tendría un rol determinante en la fisiopatología de las enfermedades cardiovasculares. La inmunidad innata es la primera línea de defensa, no específica, preprogramada para combatir agentes infecciosos. Este sistema censa la presencia de antígenos extraños a través de los receptores tipo toll (TLR) produciendo citoquinas y activando mecanismos microbicidas. Sin embargo, los TLRs también se hayan distribuidos en las células parenquimales no inmunes, jugando un importante rol tanto en la defensa como en la homeostasis de cada tejido. Durante la etapa aguda de la infección, el T. cruzi invade y se replica dentro de una amplia variedad de células y tejidos. Pero posteriormente, los parásitos son efectivamente eliminados de la mayoría de los tejidos persistiendo durante toda la vida en las células del músculo cardíaco y esquelético de los pacientes infectados. Debido a que el mantenimiento de la célula cardíaca infectada es crítica para la patogénesis de la enfermedad, los mecanismos que participan en la sobrevida de los cardiomiocitos están siendo foco de nuestro estudio. Hemos demostrado, que la infección ejerce efectos antiapoptóticos sobre células cardíacas aisladas. Nuestra hipótesis es que la inmunidad innata cardíaca estaría involucrada en el mantenimiento de la sobrevida de los miocitos así como en la defensa contra el parásito. Objetivo general: determinar la participación de la respuesta inmune innata cardíaca en el desarrollo de la enfermedad de Chagas experimental murina. Objetivos específicos: 1) Analizar el compromiso de TLRs en la respuesta anti-apoptótica y de autofagia de cardiomiocitos aislados de ratones salvajes y de ratones deficientes en TLR4, TLR2 y en MyD88, molécula adaptadora de la señalización por TLRs, sometidos a la infección con el parásito. 2) Determinar la importancia de la actividad cisteín proteasa parasitaria en el grado de infectividad y la sobrevida de cultivos primarios de ratones salvajes infectados con parásitos transgénicos que poseen disminuída o nula actividad cisteín proteasa. 3) Establecer la cinética de expresión de TLR2/TLR6, TLR4 y TLR9, factores antiapoptóticos (Bcl-2, Bcl-xL, etc.), daño cardíaco y la carga parasitaria en el tejido cardíaco de ratones infectados salvajes y/o deficientes antes mencionados. Materiales y Métodos: Los animales serán infectados i.p. con 5x103 parásitos y se determinará la cinética de expresión de los mediadores mencionados por western blot e inmunofluorescencia, la carga parasitaria será determinada por qRT-PCR. Como controles se procesarán animales inyectados con solución salina. En cultivos primarios de cardiomiocitos de ratones neonatos salvajes y deficientes infectados se estudiará la carga parasitaria, la activación de los mecanismos microbicidas (producción de óxido nítrico, metabolitos reactivos del oxígeno y del nitrógeno, ciclooxigenasa, etc.), producción de citoquinas y expresión de moléculas anti-apoptóticas (Bcl-2, Bcl-xL, Bax, etc.). Se explorará la tasa de apoptosis en cultivos deprivados de suero. La autofagia se analizará por microscopia electrónica. Cultivos controles serán mantenidos en medio o tratados con ligandos de los diferentes TLRs. Resultados preliminares sugieren que tanto TLR2 como Bcl-2 se incrementan en tejido cardíaco infectado. Esto nos lleva a profundizar en los mecanismos observados en cultivos y estudiarlos en un modelo in vivo, analizando la posible importancia que tiene la inmunidad innata cardíaca en el control del establecimiento de la infección. La comprensión de los mecanismos que mantienen la sobrevida de los cardiomiocitos y su respuesta a la infección es importante ya que el conocimiento de las bases moleculares es fundamental para el desarrollo de nuevos agentes quimioterapéuticos. Chagas disease is endemic in Central and South America and causes the most common myocarditis worldwide. We have previously reported that the cardiotrophic parasite Trypanosoma cruzi, its etiological agent, protects cardiomyocytes against apoptosis induced by growth factor deprivation activating the PI3K/Akt and MEK1/ERK signaling pathways. Recent studies have shown that local innate immunity plays a key role in initiating and coordinating homeostatic as well as defense responses in the heart. One of the mechanisms by which the innate immune system senses the presence of foreign antigens is through TLRs. The stimulation of these receptors leads to the activation and nuclear translocation of NF-kB transcription factor and the production of cytokines. Proinflammatory cytokines, in turn, appear to play a central role in the orchestration and timing of the intrinsic cardiac stress response providing, under different situations, instantaneous anti-apoptotic cytoprotective signals, which allow tissue repair and/or remodeling. The aim of the present project is to study the cardiomyocyte innate immune responses to T. cruzi infection and its role in target cell protection from apoptosis. Specific objectives: 1) Study the mechanism triggered by TLR in the anti-apoptotic response and parasite load of infected cardiomyocyte primary cultures from wild type and mice deficient in TLR2, TLR4 or MyD88. 2) Determine the effect of parasite cisteín protease activity on primary cultures from wild type mice. 3) Determine the TLR signaling-involvement in parasite load and survival indicators in deficient mice. Preliminary results showed us that cardiac-TLR2 may be involved in the anti-apoptotic effect elicited by the parasite and prompted us to establish the mechanisms triggered by the innate immunity that mediate parasite persistence within the host cell.

Expresión de las proteínas relacionadas con la proliferación celular y la apoptosis en lesiones blancas de la mucosa oral, como precursoras de la cancerización en el epitelio escamoso oral.

El cáncer de la mucosa oral es una patología muy frecuente que llega en muchos casos a conformar entre el 8 y el 10% de los tumores malignos del hombre. La baja concientización de la población general sobre el tema es un factor importante que hace que estas lesiones sean detectadas en forma tardía y cuando ya se trata de lesiones avanzadas de muy mal pronóstico. Si bien ha habido grandes avances en el diagnóstico y tratamiento de lesiones blancas de la mucosa oral como el liquen plano oral, estas lesiones siguen siendo entidades con muchos interrogantes para todos los expertos en medicina oral, sobre todo en lo referente a su proceso de aparición y a su tratamiento. Es importante la diferenciación correcta de estas lesiones, ya que el carcinoma epidermoide bucal puede aparecer también como una lesión blanca. No hay suficiente conocimiento de lo que ocurre en otras lesiones blancas de la mucosa oral no liquen plano. Para evaluar el verdadero potencial de transformación maligna de leucoplasias, liquen plano oral, reacciones liquenoides y lesiones escamosas intraepiteliales (incluídas las inducidas por acción viral) se evaluará la expresión de las proteínas relacionadas con la proliferación celular y la apoptosis en estas lesiones. Se evaluará con técnicas inmunohistoquímicas la relación de bcl-2 como marcador apoptótico en leucoplasias, liquen plano oral, lesiones escamosas intraepiteliales y carcinomas epidermoides orales, así como también en estas mismas lesiones la expresión proteica de MIB-1 (ki67), ciclina D1, p16 y p53 para valorar si las alteraciones en la expresión proteica de estos marcadores, suceden de forma secuencial a través de las distintas etapas en la cancerización del campo de cavidad oral, ya que no existe consenso en los resultados y las conclusiones obtenidas en los diferentes estudios efectuados sobre la influencia exclusiva de los marcadores apoptóticos en el desarrollo de las lesiones de algunas de las lesiones como el LPO.

Development and preclinical assessment of a bioartificial pancreas.

Transplantation of insulin secreting cells is regarded as a possible treatment for type 1 diabetes. One major difficulty in this approach is, however, that the transplanted cells are exposed to the patient's inflammatory and autoimmune environment, which originally destroyed their own beta-cells. Therefore, even if a good source of insulin-secreting cells can be identified for transplantation therapy, these cells need to be protected against these destructive influences. The aim of this project was to evaluate, using a clonal mouse beta-cell line, whether genetic engineering of protective genes could be a viable option to allow these cells to survive when transplanted into autoimmune diabetic mice. We demonstrated that transfer of the Bcl-2 anti-apoptotic gene and of several genes specifically interfering with cytokines intracellular signalling pathways, greatly improved resistance of the cells to inflammatory stresses in vitro. We further showed that these modifications did not interfere with the capacity of these cells to correct hyperglycaemia for several months in syngeneic or allogeneic streptozocin-diabetic mice. However, these cells were not protected against autoimmune destruction when transplanted into type 1 diabetic NOD mice. This suggests that in addition to inflammatory attacks by cytokines, autoimmunity very efficiently kills the transplanted cells, indicating that multiple protective mechanisms are required for efficient transplantation of insulin-secreting cells to treat type 1 diabetes.

In vivo conditional Pax4 overexpression in mature islet β-cells prevents stress-induced hyperglycemia in mice.

OBJECTIVE To establish the role of the transcription factor Pax4 in pancreatic islet expansion and survival in response to physiological stress and its impact on glucose metabolism, we generated transgenic mice conditionally and selectively overexpressing Pax4 or a diabetes-linked mutant variant (Pax4R129 W) in β-cells. RESEARCH DESIGN AND METHODS Glucose homeostasis and β-cell death and proliferation were assessed in Pax4- or Pax4R129 W-overexpressing transgenic animals challenged with or without streptozotocin. Isolated transgenic islets were also exposed to cytokines, and apoptosis was evaluated by DNA fragmentation or cytochrome C release. The expression profiles of proliferation and apoptotic genes and β-cell markers were studied by immunohistochemistry and quantitative RT-PCR. RESULTS Pax4 but not Pax4R129 W protected animals against streptozotocin-induced hyperglycemia and isolated islets from cytokine-mediated β-cell apoptosis. Cytochrome C release was abrogated in Pax4 islets treated with cytokines. Interleukin-1β transcript levels were suppressed in Pax4 islets, whereas they were increased along with NOS2 in Pax4R129 W islets. Bcl-2, Cdk4, and c-myc expression levels were increased in Pax4 islets while MafA, insulin, and GLUT2 transcript levels were suppressed in both animal models. Long-term Pax4 expression promoted proliferation of a Pdx1-positive cell subpopulation while impeding insulin secretion. Suppression of Pax4 rescued this defect with a concomitant increase in pancreatic insulin content. CONCLUSIONS Pax4 protects adult islets from stress-induced apoptosis by suppressing selective nuclear factor-κB target genes while increasing Bcl-2 levels. Furthermore, it promotes dedifferentiation and proliferation of β-cells through MafA repression, with a concomitant increase in Cdk4 and c-myc expression.

Transcriptional regulation of CD4 gene expression by T cell factor-1/beta-catenin pathway.

By interacting with MHC class II molecules, CD4 facilitates lineage development as well as activation of Th cells. Expression of physiological levels of CD4 requires a proximal CD4 enhancer to stimulate basic CD4 promoter activity. T cell factor (TCF)-1/beta-catenin pathway has previously been shown to regulate thymocyte survival via up-regulating antiapoptotic molecule Bcl-xL. By both loss and gain of function studies, in this study we show additional function of TCF-1/beta-catenin pathway in the regulation of CD4 expression in vivo. Mice deficient in TCF-1 displayed significantly reduced protein and mRNA levels of CD4 in CD4+ CD8+ double-positive (DP) thymocytes. A transgene encoding Bcl-2 restored survival but not CD4 levels of TCF-1(-/-) DP cells. Thus, TCF-1-regulated survival and CD4 expression are two separate events. In contrast, CD4 levels were restored on DP TCF-1(-/-) cells by transgenic expression of a wild-type TCF-1, but not a truncated TCF-1 that lacks a domain required for interacting with beta-catenin. Furthermore, forced expression of a stabilized beta-catenin, a coactivator of TCF-1, resulted in up-regulation of CD4. TCF-1 or stabilized beta-catenin greatly stimulated activity of a CD4 reporter gene driven by a basic CD4 promoter and the CD4 enhancer. However, mutation of a potential TCF binding site located within the enhancer abrogated TCF-1 and beta-catenin-mediated activation of CD4 reporter. Finally, recruitment of TCF-1 to CD4 enhancer was detected in wild-type but not TCF-1 null mice by chromatin-immunoprecipitation analysis. Thus, our results demonstrated that TCF/beta-catenin pathway enhances CD4 expression in vivo by recruiting TCF-1 to stimulate CD4 enhancer activity.

Effect of antiretroviral therapy on apoptosis markers and morphology in peripheral lymph nodes of HIV-infected individuals.

BACKGROUND: CD4+ T cell depletion and destruction and the involution of the lymphoid tissue are hallmarks of HIV infection. Although the underlying mechanisms are still unclear, apoptosis appears to play a central role. The objective of this study was to investigate the effect of antiretroviral therapy on the lymph node tissue, particularly with respect to morphology and apoptosis. PATIENTS AND METHODS: Between 1997 and 1999, two inguinal lymph nodes were excised from 31 previously untreated individuals who were in an early stage of HIV infection, the first one prior to treatment and the second after 16 to 20 months of treatment. Paraffin sections were investigated for lymph node architecture, distribution of cellular and viral markers, apoptosis, and expression of apoptotic key molecules which indirectly reflect apoptotic processes. RESULTS: After 16-20 months of antiretroviral therapy, a significant decrease in highly activated HIV-driven immune response was observed in the lymph node tissue as a marked reduction in follicular hyperplasia, a normalization of the follicular dendritic cell network, a significant increase in the number of CD4+ T cells, and a significant decrease in the number of CD8+ T cells. The expression of several proapoptotic (Fas, TRAIL, and active caspase 3) and antiapoptotic (Bcl-2 and IL-7Ralpha) molecules that were reconstituted in the tissues during therapy resembled their expression in lymph nodes of HIV-negative individuals. Limitations of the study are (a) the lack of untreated patients in the late stages, (b) for ethical reasons, the lack of a control group with untreated patients, and (c) for methodological reasons, the restriction of sequential measurements of apotpotic markers to one-third of the patients. CONCLUSION: Antiretroviral therapy initiated in the early stages in HIV infection may halt the irreversible destruction of the lymph node tissue and may partially normalize apoptotic processes.

The beta-catenin--TCF-1 pathway ensures CD4(+)CD8(+) thymocyte survival.

The association of trans-acting T cell factors (TCFs) or lymphoid enhancer factor 1 (LEF-1) with their coactivator beta-catenin mediates transient transcriptional responses to extracellular Wnt signals. We show here that T cell maturation depends on the presence of the beta-catenin--binding domain in TCF-1. This domain is necessary to mediate the survival of immature CD4(+)CD8(+) double-positive (DP) thymocytes. Accelerated spontaneous thymocyte death in the absence of TCF-1 correlates with aberrantly low expression of the anti-apoptotic protein Bcl-x(L). Increasing anti-apoptotic effectors in thymocytes by the use of a Bcl-2 transgene rescued TCF-1-deficient DP thymocytes from apoptosis. Thus, TCF-1, upon association with beta-catenin, transiently ensures the survival of immature T cells, which enables them to generate and edit T cell receptor (TCR) alpha chains and attempt TCR-mediated positive selection.

Destruction of conditional insulinoma cell lines in NOD mice: a role for autoimmunity.

AIMS/HYPOTHESIS: betaTC-tet (H2(k)) is a conditional insulinoma cell line derived from transgenic mice expressing a tetracycline-regulated oncogene. Transgenic expression of several proteins implicated in the apoptotic pathways increase the resistance of betaTC-tet cells in vitro. We tested in vivo the sensitivity of the cells to rejection and the protective effect of genetic alterations in NOD mice. METHODS: betaTC-tet cells and genetically engineered lines expressing Bcl-2 (CDM3D), a dominant negative mutant of MyD88 or SOCS-1 were transplanted in diabetic female NOD mice or in male NOD mice with diabetes induced by high-dose streptozotocin. Survival of functional cell grafts in NOD-scid mice was also analyzed after transfer of splenocytes from diabetic NOD mice. Autoreactive T-cell hybridomas and splenocytes from diabetic NOD mice were stimulated by betaTC-tet cells. RESULTS: betaTC-tet cells and genetically engineered cell lines were all similarly rejected in diabetic NOD mice and in NOD-scid mice after splenocyte transfer. In 3- to 6-week-old male NOD mice treated with high-dose streptozotocin, the cells temporarily survived, in contrast with C57BL/6 mice treated with high-dose streptozotocin (indefinite survival) and untreated 3- to 6-week-old male NOD mice (rejection). The protective effect of high-dose streptozotocin was lost in older male NOD mice. betaTC-tet cells did not stimulate autoreactive T-cell hybridomas, but induced IL-2 secretion by splenocytes from diabetic NOD mice. CONCLUSION/INTERPRETATION: The autoimmune process seems to play an important role in the destruction of betaTC-tet cells in NOD mice. Genetic manipulations intended at increasing the resistance of beta cells were inefficient. Similar approaches should be tested in vivo as well as in vitro. High dose streptozotocin influences immune rejection and should be used with caution.

Children Order (PDF 508 KB)

The Children Order series of regulations and guidance consists of the following: Volume 1: Court Orders and other Legal Issues Volume 2: Family Support, Child Minding and Day Care Volume 3: Family Placements and Private Fostering Volume 4: Residential Care Volume 5: Children with a Disability Volume 6: Co-operating to Protect Children Volume 7: Schools Accommodating Child åÊ

Growing up in Ireland. Key findings: infant cohort (at 3 years).

 The latest results from the study paint a picture of how these families are faring across a range of areas in their lives including their health, family life and financial and economic circumstances. In general the findings show that three-year-olds in Ireland are in good health with a few notable public health and related issues (including overweight and obesity), there is overall stability in family structures over the short term and that the recession has had a substantial effect on families with young children over the last number of years. These are the first longitudinal findings from the study. The first wave of fieldwork with the families of the Infant Cohort included approximately 11,100 nine-month-olds, their parents and carers. Interviews began in September 2008 and were completed in March 2009. Interviews for the second round of interviews with this cohort took place between January and August 2011. A total of 90% of the original sample of nine-month-olds were successfully re-interviewed. (A full download of the results released today, presented in three briefing documents can be found by clicking here. Key findings include: Health â?¢ Most of the children were described as being in good health; 75% were rated as very healthy and a further 23% were rated as healthy, but a few minor problems. Girls were more likely to be reported as very healthy (78%) compared with boys (72%). â?¢ One in four or almost one quarter of three-year-old children were overweight (19%) or obese (6%). â?¢ Childrenâ?Ts weight was related to household social class. 5% of children in families in the professional/managerial group were classified as obese at three years of age compared with 9% of those in the most disadvantaged social class group. However, at least one-fifth of children in every social class were overweight. â?¢ Childrens consumption of energy-dense foods such as crisps, sweets, chips, and non-diet fizzy drinks increased as parental education fell. 63% of children whose mother had a lower secondary education or less ate at least one portion of crisps compared with 36% of those from degree-level backgrounds, although consumption of biscuits/chocolates was over 70% for both groups of children. â?¢ Two-thirds (66%) of three-year-olds had received at least one course of antibiotics in the 12 months preceding the interview. Children with a full medical card (35% of the sample) or a GP-only medical card (5% of the sample) were more likely to have received a course of antibiotics than â?¢ Children with a full medical card received a higher number of antibiotic courses on average (2.6) compared with those without a medical card (2.1). â?¢ Just under 16% of three-year-old children were reported as having at least one longstanding illness, condition or disability. The most commonly reported illness types included Asthma (5.8%), Eczema/Skin allergies (3.9%) and Food/digestive allergies (1.2%) Family Life and Childcare â?¢ While the overall distribution of family structure was stable, there have been transitions from one-parent families to two-parent families and vice-versa over the 27 months between interview â?" approximately 2 to 3 percent in each direction. â?¢ 50% of three year olds were in some form of non-parental childcare for eight or more hours a week. The most common form used was centre-based childcare which almost tripled between nine months and three years, from 11% to 30%. â?¢ A similar percentage of grandparents were caring for children at both nine months and three years, 12% and 11% respectively. A total of 10% of three-year-olds were being minded by a childminder, an increase of 3 percentage points from when the children were nine months of age. â?¢ Children who were in some form of non-parental childcare were spending an average of 23 hours a week in their main type of childcare. â?¢ At time of interview the vast majority of mothers reported that they had regular contact with the Study Childâ?Ts grandparents (91%). In offering support to parents, grandparents were most likely to babysit (50%), and buy clothes (40%) at least on a monthly basis. One-parent families were more likely than two-parent families to receive financial support from grandparents with just under one-third (66%) of one-parent families receiving financial support from grandparents at least once every three months. â?¢ The most frequently used discipline technique was â?~discussing or explaining why the behaviour was wrongâ?T, with 63% of mothers saying they always did this. â?¢ 12% of mothers said they used â?~smackingâ?T as a form of discipline now and again and less than 1% used â?~smackingâ?T as a form of discipline more frequently. Over half reported that they never smacked the Study Child. Financial and Economic Circumstances â?¢ Just over half (53%) of mothers of three-year-olds worked outside the home, 38% said they were on home duties and 6% said they were unemployed. â?¢ The biggest change in terms of the work status of three-year-oldsâ?T parents was an increase in the percentage of unemployed fathers â?" 6% when the child was nine months rising to almost 14% when s/he was three years of age. â?¢ 61% of families of three-year-olds reported experiencing difficulties in making â?~ends meetâ?T. This was a substantial increase from 44% in the first round of interviews when the children were nine-months-old. â?¢ Almost two thirds (63%) of all families with three-year-olds reported that the recession had had a very significant or significant effect on them. â?¢ The most frequently recorded effects were: a reduction in wages (63%); canâ?Tt afford luxuries (54%), social welfare reduction (53%) and canâ?Tt afford/cut back on basics (32%). Growing Up in Ireland is a Government funded study tracking the development of two nationally representative cohorts of children: an Infant Cohort which was interviewed initially at nine months and subsequently at three years of age; and a Child Cohort which was interviewed initially at nine years and subsequently at 13 years of age. The study is being conducted by a consortium of researchers led by the Economic and Social Research Institute (ESRI) and Trinity College Dublin. For Further Information Please Contact: Jillian Heffernan Communications Officer, Growing Up in Ireland Tel: 01 896 3378 Mobile: 087 9016880This resource was contributed to our repository by the National Documentation Centre on Drug Use.

Activated peripheral lymphocytes with increased expression of cell adhesion molecules and cytotoxic markers are associated with dengue fever disease

The immune mechanisms involved in dengue fever and dengue hemorrhagic/dengue shock syndrome are not well understood. The ex vivo activation status of immune cells during the dengue disease in patients was examined. CD4and CD8 T cells were reduced during the acute phase. Interestingly, CD8 T cells co-expressing activation marker HLA-DR, Q, P, and cytolytic granule protein-Tia-1 were significantly higher in dengue patients than in controls. Detection of adhesion molecules indicated that in dengue patients the majority of T cells (CD4 and CD8) express the activation/memory phenotype, characterized as CD44HIGH and lack the expression of the naïve cell marker, CD62L LOW. Also, the levels of T cells co-expressing ICAM-1 (CD54), VLA-4, and LFA-1 (CD11a) were significantly increased. CD8 T lymphocytes expressed predominantly low levels of anti-apoptotic molecule Bcl-2 in the acute phase, possibly leading to the exhibition of a phenotype of activated/effector cells. Circulating levels of IL-18, TGF-b1 and sICAM-1 were significantly elevated in dengue patients. Early activation events occur during acute dengue infection which might contribute to viral clearance. Differences in expression of adhesion molecules among CD4 and CD8 T cells might underlie the selective extravasation of these subsets from blood circulation into lymphoid organs and/or tissues. In addition, activated CD8 T cells would be more susceptible to apoptosis as shown by the alteration in Bcl-2 expression. Cytokines such as IL-18, TGF-b1, and sICAM-1 may be contributing by either stimulating or suppressing the adaptative immune response, during dengue infection, thereby perhaps establishing a relationship with disease severity.

Delay in maturation of the submandibular gland in Chagas disease correlates with lower DNA synthesis

It has been demonstrated that the acute phase of Trypanosoma cruzi infection promotes several changes in the oral glands. The present study examined whether T. cruzi modulates the expression of host cell apoptotic or mitotic pathway genes. Rats were infected with T. cruzi then sacrificed after 18, 32, 64 or 97 days, after which the submandibular glands were analyzed by immunohistochemistry. Immunohistochemical analyses using an anti-bromodeoxyuridine antibody showed that, during acute T. cruzi infection, DNA synthesizing cells in rat submandibular glands were lower than in non-infected animals (p < 0.05). However, after 64 days of infection (chronic phase), the number of immunolabeled cells are similar in both groups. However, immunohistochemical analysis of Fas and Bcl-2 expression did not find any difference between infected and non-infected animals in both the acute and chronic stages. These findings suggest that the delay in ductal maturation observed at the acute phase of Chagas disease is correlated with lower expression of DNA synthesis genes, but not apoptotic genes.

Induction of apoptosis in A549 pulmonary cells by two Paracoccidioides brasiliensis samples

Paracoccidioidomycosis presents a variety of clinical manifestations and Paracoccidioides brasiliensis can reach many tissues, most importantly the lungs. The ability of the pathogen to interact with host surface structures is essential to its virulence. The interaction between P. brasiliensis and epithelial cells has been studied, with particular emphasis on the induction of apoptosis. To investigate the expression of different apoptosis-inducing pathways in human A549 cells, we infected these cells with P. brasiliensis Pb18SP (subcultured) and 18R (recently isolated from cell culture and showing a high adhesion pattern) samples in vitro. The expressions of Bcl-2, Bak and caspase 3 were analysed by flow cytometry and DNA fragmentation using the TUNEL technique. Apoptosis of human A549 cells was induced by P. brasiliensis in a sample and time-dependent manner. Using an in vitro model, our data demonstrates that caspase 3, Bak, Bcl-2 and DNA fragmentation mediate P. brasiliensis-induced apoptosis in A549 cells. The overall mechanism is a complex process, which may involve several signal transduction pathways. These findings could partially explain the efficient behaviour of this fungus in promoting tissue infection and/or blood dissemination.