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.

The proliferating cell nuclear antigen index in breast carcinomas does not correlate with mitotic index and estrogen receptor immunoreactivity

To investigate the expression of a marker of cell proliferation (PCNA/Cyclin) and its putative relationship with histological grading, mitotic index and estrogen receptor immunoreactivity, we studied twenty-seven cases of invasive breast carcinoma in formalin-fixed, paraffin-embedded tissue sections. The PCNA and estrogen receptor were detected by the PC10 and H222 monoclonal antibodies respectively, using an avidin-biotin-pernxidase method. The median value of PCNA index was 20.9% with a range from 1.4 to 84.2%. We did not find any significant relationship between PCNA index anti the histological grading, mitotic index and estrogen receptor immunoreactivity. We conclude that PCNA detected by the monoclonal antibody PC10 in formalin-fixed material looks at present unrealiable as a proliferation marker in breast carcinoma.

DETECTION OF CELL-PROLIFERATION IN TISSUE-SECTIONS

1. Cell proliferation is of interest since abnormal cell proliferation appears to be a precursor of tumorigenesis and also because the quantitative description of cell proliferation in tumors can be used to predict the biological behavior of a particular neoplasia.2. Them am several reliable methods of studying cell proliferation in tissues. One of the most important is the detection of the Ki67 defined antigen in frozen sections. The number of cells expressing Ki67 correlates with histological grades of tumors and can also be predictive of clinical outcome. The Ki67 can be localized in tissue sections using monoclonal antibodies in association with the immunoperoxidase technique.3. Proliferating cell nuclear antigen (PCNA) is a component of DNA polymerase-delta and is another important cell proliferation marker manifesting a striking increase in concentration during the S phase of the cell cycle. 19A2 and PC10 are two different monoclonal antibodies which can be employed to detect PCNA in paraffin-embedded tissues.4. Molecular biology has also been making a great contribution to the study of cell proliferation. The most recent innovation in tissue identification of proliferating cells is the use of in situ hybridization for the localization of histone H3 and/or H4 mRNA. H3 mRNA-positive cells appear to be present in basal cells of the skin and in crypt cells of the intestine which are sites with high proliferation rate.

Influência do clareamento dental na cor, translucidez e fluorescência do esmalte e dentina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da alteração de cor, difusão de peróxido de hidrogênio e citotoxicidade trans-amelodentinária causadas por diferentes técnicas de clareamento dental

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Age determination of sediment cores from the Skagerrak

Paleoceanographic and paleoenvironmental interpretations based on foraminifera, sedimentary data, radiocarbon dates, and stable isotope measurements were derived from two sections in the Skagerrak: a 115-m-thick Holocene marine section drilled onshore at Skagen near the northernmost tip of Jutland, Denmark, and a 9-m piston core from the Skagerrak, north of Skagen. The foraminiferal data show that arctic-subarctic environments in the deep Skagerrak-Kattegat area were succeeded by boreal conditions at 9.6 ka. This was a result of northward migration of the Atlantic polar front and inflow of warm Atlantic water into the area through the Norwegian Channel. A gradual warming of the water masses after 9.6 ka is indicated by the data. Rare foraminifera and high sedimentation rates are found between approximately 8.6 ka and 7.6 ka at both core locations. The modern foraminiferal assemblages of the area were fully established at 7.6 ka indicating that the modern circulation pattern in the Skagerrak-Kattegat after the opening of the English Channel and the Danish Straits was not established before this date. At 5.5 ka a sudden change to coarser sediments (higher-energy environments) and the appearance of the foraminifer Eoeponidella laesoeensis is recorded in the Skagen core. This indicates a rapid change in the hydrography reflecting altered meteorological and hydrographic conditions in the Skagerrak-Kattegat, including a strengthening of the Jutland Current and increased inflow of North Sea water into the Kattegat. The event is interpreted as a response to cooling at the end of the Holocene climatic optimum in late Atlantic time and possibly reflects a rapid cooling event of North Atlantic surface water masses.

(Table 2) Radiocarbon ages of selected southern Baja California sediment cores

Piston, gravity, and multicores as well as hydrographic data were collected along the Pacific margin of Baja California to reconstruct past variations in the intensity of the oxygen-minimum zone (OMZ). Gravity cores collected from within the OMZ north of 24°N did not contain laminated surface sediments even though bottom water oxygen (BWO) concentrations were close to 5 µmol/kg. However, many of the cores collected south of 24°N did contain millimeter- to centimeter-scale, brown to black laminations in Holocene and older sediments but not in sediments deposited during the Last Glacial Maximum. In addition to the dark laminations, Holocene sediments in Soledad Basin, silled at 290 m, also contain white coccolith laminae that probably represent individual blooms. Two open margin cores from 430 and 700 m depth that were selected for detailed radiocarbon dating show distinct transitions from bioturbated glacial sediment to laminated Holocene sediment occurring at 12.9 and 11.5 ka, respectively. The transition is delayed and more gradual (11.3-10.0 ka) in another dated core from Soledad Basin. The observations indicate that bottom-water oxygen concentrations dropped below a threshold for the preservation of laminations at different times or that a synchronous hydrographic change left an asynchronous sedimentary imprint due to local factors. With the caveat that laminated sections should therefore not be correlated without independent age control, the pattern of older sequences of laminations along the North American western margin reported by this and previous studies suggests that multiple patterns of regional productivity and ventilation prevailed over the past 60 kyr.