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A 78 year-old female patient underwent a total mastectomy with axillary lymph node dissection for a primary breast osteosarcoma. Microscopically the tumor was identical to grade II skeletal osteosarcoma. Immunohistochemically no reactivity was detected, either for the epithelial markers EMA, AE1/AE3, CK8, 18, 19, or for HER-2/neu, estrogen and progesterone receptors, as well as fluorescent IN SITU hybridization for HER-2/neu. The diagnosis of this tumor fulfills certain clinicopathological criteria. Mammary osteosarcoma is usually developed in phyllodes tumors or carcinosarcomas of the breast as a result of metaplasia of the epithelial component. This rare tumor of the breast is occasionally associated with prior radiation therapy or well documented trauma. Mammary osteosarcoma is a biologically aggressive neoplasm with a 38% five-year survival rate. Surgical resection is the most effective therapy to date. Adjuvant treatment -chemotherapy or radiotherapy- has shown no clear benefit. An extensive review of the literature is also presented.

1. Introducció:El càncer de mama té un enorme impacte en la salut de la dona degut a la seva elevada incidència, mortalitat i prevalença. A Catalunya representa també la primera causa de mort per càncer a les dones. Segons dades recents aportades per l'actualització del metaanàlisi sobre tractament complementari a la cirurgia, la quimioteràpia adjuvant aporta un benefici significatiu sobre la recidiva i supervivència de les pacients amb càncer de mama. Aquest benefici està influenciat pel risc de recidiva que presentin aquestes pacients en el moment del diagnòstic. Les pacients amb extensa afectació ganglionar axil·lar presenten un elevat risc de recidiva, tot i no objectivar-se afectació metastàsica en el moment del diagnòstic. El càncer de mama presenta una gran heterogeneïtat en el seu comportament. Aprofundir en l'estudi de la biologia tumoral, delimitant la implicació de cada factor pronòstic i predictiu de risc de recidiva i mort, ha de permetre millorar la seva curabilitat amb la menor morbilitat possible.L'esquema de quimioteràpia adriamicina (A) seguit de CMF (ciclofosfamida, metotrexat i fluorouracil) s'ha mostrat superior a esquemes amb CMF i a pautes amb altes dosis de quimioteràpia. No és disposa encara d'estudis definitius que defineixin el seu paper front a esquemes amb taxans.2. Pacients i mètodesCent seixanta-tres pacients diagnosticades de càncer de mama entre els anys 1992 i 1997, amb afectació de més de tres ganglis axil·lars i sense metàstasis a distància han estat tractades amb l'esquema de quimioteràpia A  CMF (quatre cicles de adriamicina seguits de vuit cicles de CMF cada 21 dies). La radioteràpia sobre mama o paret amb àrees ganglionars es va iniciar desprès de l'adriamicina i simultàniament a CMF. Una tercera part de les pacients amb receptors hormonal positius van rebre tractament hormonal amb tamoxifè al finalitzar la quimioteràpia i durant cinc anys.En l'estudi dels factors pronòstics s'han inclòs l'edat, l'estat menstrual, la mida tumoral, el grau histològic, el nombre de ganglis axil·lars afectes, els receptors d'estrògens i progesterona, la sobreexpressió de l'oncogèn HER-2 i l'amenorrea quimioinduïda.El seguiment i anàlisi de les dades s'ha efectuat de forma centralitzada i prospectiva. Per l'estudi de l'eficàcia sobre la recidiva i la supervivència s'ha utilitzat la supervivència lliure de malaltia (SLLM) i la supervivència global (SG). 3. ResultatsAmb una mediana de seguiment en el moment de l'anàlisi dels resultats de 86 mesos, s'ha observat una SLLM del 68 % i una SG del 78 %. En poques pacients s'han observat toxicitats agudes serioses, sense observar-se morts tòxiques com a conseqüència del tractament. La cardiotoxicitat clínica atribuïble ha estat del 1 % i fins al moment una pacient ha estat diagnosticada d'un procés leucèmic.En l'anàlisi dels factor pronòstics, el nombre de ganglis afectes, la mida tumoral i el grau histològic s'han mostrat com a paràmetres amb una influència significativa en l'avaluació multivariada del conjunt de pacients. Quan s'ha analitzat només les pacients premenopàusiques en el moment de l'inclusió, l'amenorrea s'esdevé un factor amb significació pronòstica independent.La sobreexpressió d'HER-2 i els RH no aporten informació pronòstica significativa en l'anàlisi multivariat.Quan s'estableixen comparacions indirectes entre els nostres resultats i els obtinguts per altres pautes de tractament quimioteràpia i per aquest subgrup de pacients, cap dels estudis publicats fins l'actualitat mostra taxes de SLLM i SG clarament superiors.4. Discussió i conclusionsLa pauta de tractament A  CMF, dins d'un plantejament de tractament combinat amb radioteràpia, presenta una efectivitat remarcable, en termes de SLLM i SG, i en el grup de pacients d'alt risc de recidiva.La pauta estudiada és perfectament aplicable en la pràctica assistencial, donat el seu compliment, la seva acceptable toxicitat aguda i tardana i la seva aplicabilitat en règim multiinstitucional.La mida tumoral, el grau histològic i el nombre de ganglis afectes mostren una influència esperada significativa e independent sobre la SLLM i la SG. L'impacte clarament significatiu de l'amenorrea secundària a la quimioteràpia sobre la recidiva i la mort i en les pacients premenopàusiques, obliga a considerar la supressió ovàrica de les pacients que no presentin amenorrea desprès de la quimioteràpia.La sobreexpressió de l'oncogèn HER-2 no confereix un pronòstic advers en les pacients que l'expressen i són tractades amb A  CMF; a diferència de altres tipus de quimioteràpia. Aquest és un fet a considera quan es consideri l'inclusió del tractament específic amb trastuzumab per les pacients amb HER-2 positiu.Els receptors hormonals tampoc afecten significativament sobre l'eficàcia de la pauta A  CMF. Per tant, i a diferència d'esquemes amb paclitaxel, també les pacients amb receptors hormonals positius semblen beneficiar-se de la pauta estudiada.Un subgrup de pacient amb tumors ben o moderadament diferenciats, tot i l'extensa afectació ganglionar axil·lar, presenten taxes de supervivència molt favorables, suggerint que el règim de tractament A  CMF pot ser especialment actiu i recomanable per aquest subgrup de pacients., We sought to evaluate outcomes and the most likely prognostic factors associated with the use of sequential doxorubicin + cyclophosphamide-methotrexate-fluorouracil administered prior to scheduled radiation therapy, in non-selected consecutive patients (n=163) with 4 positive nodes. Variables included in statistical analyses included age, menopausal status, tumour size, histology grade, number of positive nodes (4 to 9 or 10), oestrogen and progesterone receptors (ER, PR), HER-2 status and chemotherapy-induced amenorrhoea. With a median follow-up of 86 months, the results showed disease-free survival (DFS) at 5 years was 68% and overall survival (OS) was 78% and no toxic deaths. Number of positive nodes, tumour size, histology grade and amenorrhoea showed prognostic significance for DFS and OS. ER and PR reached significance only for OS. HER-2 positive status did not predict poorer DFS or OS. Tumor size > 2 cm, 10+ positive nodes and grade 3 tumours predicted DFS and OS in multivariate analysis. Amenorrhoea reach prognostic significance for DFS and OS in premenopausal patients. We conclude that DOXCMF is an effective and safe regimen. Number of positive axillary nodes, histologic grade and tumor size , but not HER2 status, are predictive of outcome. Amenorrhoea favour DFS and OS in premenopausal patients.

Introducción: Se estima que en el año 2000 se diagnosticaron más de 10 millones de cánceres colorectales en el mundo y más de 6 millones murieron por esta causa. En Europa se ha observado un aumento de la incidencia desde los años 60 así como una mejora de la supervivencia. En su etiología, además de una causa genética, se ha evidenciado el estilo de vida occidentalizado y, dentro de él, la dieta como principal factor ambiental. Está en debate la mejor estrategia de cribado a nivel poblacional para este cáncer. El tratamiento de elección es la cirugía. Desde 1990 está demostrado el beneficio de la quimioterapia y radioterapia adyuvantes. Asimismo, la poliquimioterapia en la enfermedad avanzada aumenta la supervivencia. Objetivo:Conocer la incidencia y la supervivencia poblacional del cáncer colorectal en Tarragona por género, edad, sublocalización, histología y estadio.Estudiar la evolución temporal de estos indicadores.Comparar los resultados obtenidos con los de otros registros de cáncer de base poblacional.Construir un patrón epidemiológico del cáncer colorectal para la provinvia de Tarragona.Material y métodos:Se trata de un estudio epidemiológico poblacional descriptivo del cáncer colorectal a partir de la información del Registro de Cáncer de Tarragona desde el año 1980 al 1998. Se estiman las tasas de incidencia brutas, ajustadas por edad a la población mundial estándar, truncadas y acumuladas así como las tasas específicas por edad, global y por género, edad, sublocalización, histología y estadio. Se realiza un análisis de la tendencia temporal utilizando los modelos de regresión de Poisson y de Join Point, y se estiman los porcentajes anuales de cambio y su significación estadística.En cuanto a la supervivencia, se calcula la supervivencia observada global (método no paramétrico de Kaplan y Meier) y la supervivencia relativa global (método de Ederer) por género, edad, sublocalización, histología y estadio. Se determina la asociación de diversas variables con el tiempo de supervivencia utilizando el método de riesgos proporcionales de Cox, y se estudia la evolución de la supervivencia desde 1985 hasta 1998 por períodos. Los resultados de incidencia se han comparado con los publicados por diferentes registros de cáncer de todo el mundo y los de supervivencia con los de registros europeos.Resultados:La tasa bruta de incidencia es de 56,5 para ambos géneros juntos. Las tasas ajustadas son de 61,7 en hombres y 51,3 en mujeres. Las sublocalizaciones por orden de frecuencia son: recto, colon izquierdo y colon derecho en hombres, y colon izquierdo, colon derecho y recto en mujeres. La distribución porcentual por estadios (clasificación de Dukes) es: A 13%, B 27%, C 27% y D 22%. La incidencia ha aumentado un 3,3% anual en hombres y un 2,5% anual en mujeres de forma constante a lo largo de todo en período. El cáncer de colon derecho es el que más aumenta en hombres (6,5% anual) y en mujeres (5,2% anual).La supervivencia observada a 5 años es de 40,4% y la relativa de 49,0% (47,6% en hombres y 50,7% en mujeres). Por edades, las supervivencias relativas a 5 años son: 60% entre los de 35 y 54 años, 53% entre los de 55 y 74 años, y 45% en los mayores de 74 años. Por estadios, la supervivencia a 5 años es: A 90%, B 68%, C 47% y D 3%. El estadio es el factor pronóstico más importante, delante de la edad. La supervivencia relativa a 5 años pasó de 42% entre los casos diagnosticados en el periodo 1985-89 a 49% en el periodo 1990-94.Conclusiones:En Tarragona el cáncer colorectal es el cáncer más frecuente en ambos géneros juntos y presenta una incidencia media-baja con respecto a otros registros de Europa pero con una tendencia rápidamente ascendente y continua, muy probablemente debida a la adopción de un estilo de vida occidentalizado y, sobre todo, al cambio de la dieta. Las tasas de supervivencia alcanzadas se pueden considerar medio-altas en el contexto europeo, y han aumentado de forma similar a la de la mayoría de países de Europa., Introduction: It was estimated, for year 2000, that more than ten million people would be diagnosed of colorectal cancers in the world and more than six million people would die of it. In Europe, the incidence of colorectal cancer has been growing since 1960 and the survival has also improved. In its etiology, next to a genetic base, is the westernized way of life the most important factor (mainly a diet rich in proteins and fats and poor in fruits and vegetables). The adenoma is the precancerous lesion. There's controversy about the best population screening strategy for this cancer. The standard treatment is surgery. Since 1990, is the benefit of adjuvant therapies (chemotherapy and radiotherapie) confirmed. Also, the polichemotherapie has shown a survival benefit for the patients with advanced disease. Objetives:To know the incidence and survival, at a population level, of colorectal cancer in Tarragona according to gender, age, subsite of origin, histology and stage. To study the temporal trends of these sanitary indicators: incidence and survival.To compare our results with the results of other population-based cancer registries of the world.To describe an epidemiological pattern for colorectal cancer in Tarragona.Methods:Descriptive epidemiological study of colorectal cancer in Tarragona between 1980 and 1998. The data come from the population-based Cancer Registry of Tarragona. Incidence rates: normal, rates and standard rates, adjusted to the world population (Segi, 1950) have been calculated, according to gender, age, subsite of origin, histology and stage. For the temporal trends study (1980-1998) we used the Poisson model and the joinpoint model and calculated the anual percentage of change for each category. The data have been compared with the data of other registries published in Cancer Incidence Five Continents.Survival rates: observed survival rate (Kaplan and Meier model) and relative survival rate (Ederer model) have been calculated, according to gender, age, subsite of origin, histology and stage. The Cox model was uded to study the association between the different variables and the time of survival for the years 1996 and 1997. A study of the evolution of survival rates was also made (1985-1998). The data have been compared with the data of other registries published in EUROCARE-3 and the data of the SEER Programm in USA..Resultados:The normal incidence rate is 56,5 cases/100.000 inhabitants*year for both genders together. The adjusted incidence rate is 61,7 for men and 51,3 for women. The most frequent subsite of origin is left colon cancer. After this comes rectal cancer and right colon cancer. The distribution according to stages (Dukes classification) is: A (13%), B (27%), C (27%), D (22%). The incidence rates have grown 3,3% each year in men and 2,5% in women. The trend is constant between 1980 and 1998. The subsite of origin with a fastest growth is right colon cancer (6,5% in men and 5,2% in women). The 5-years observed survival rate is 40,4% and the 5-year relative survival rate is 49%. According to age-groups, the 5-year relative survival rates are: 60% for people between 35-55 years old; 53% for the ones between 55-75 years old and 45% for the people older than 74 years old. According to stages, the 5-year relative survival rates are: A (90%), B (68%), C (47%) and D (3%). The stage is the most important prognostic factor, next to the age. Between 1985 and 1994 the 5-year relative survival rate has improved from 42% to 49%.Conclusions:Colorectal cancer is the most frequent cancer in Tarragona for both genders together and the incidence rates are in the average in comparison to other European countries. The incidence has grown fast and constant. The cause of this change is probably the most westernized way of life with the time (for example: the changes in the diet). The survival rates are also in the average of the European countries and have grow from 1985 till 1994, like in the other countries, probably because of improvements in the therapies.

En la especie canina, la rodilla es la articulación que más frecuentemente presenta alteraciones, siendo las patologías de los ligamentos y meniscos las más comunes, asociadas a cambios degenerativos articulares secundarios. Para evaluar las estructuras que componen la rodilla y poder diagnosticar dichas alteraciones podemos emplear numerosas técnicas de diagnóstico por imagen como la artrografia, ecografía así como estudios de imagen por resonancia magnética (IRM). La artrografia de la rodilla, es una técnica fácil de realizar y económica, mientras que el resto de métodos son más sofisticados y necesitan una mayor inversión para llevarlos a cabo. Los estudios artrográficos se han empleado durante años en medicina humana, especialmente para el diagnóstico de lesiones en meniscos y ligamentos cruzados. Sin embargo, en veterinaria existe poca información sobre la técnica de la artrografia y anatomía radiográfica en la articulación de la rodilla porque comúnmente se emplea para el diagnóstico de las osteocondritis disecante de la articulación del hombro. Otro medio de diagnóstico de utilidad en el diagnóstico de anomalías en cartílagos, meniscos, músculos, tendones y ligamentos, así como de artropatías y neoplasias es la ecografía. La capacidad de los ultrasonidos para ofrecer imágenes de tejidos blandos intraarticulares permite obtener información que puede ser empleada de forma adicional a la imagen radiográfica. Por último, el estudio por imagen de resonancia magnética (IRM) es una de las técnicas más recientes de diagnóstico por imagen, aunque su empleo actual en medicina veterinaria es aún limitado. Debido a su capacidad de visualizar estructuras intraarticulares es la modalidad de imagen preferida para el estudio del cartílago articular, los meniscos y los ligamentos de las articulaciones sinoviales, además al contrario que la artrografia, la IRM es una técnica no invasiva y no utiliza radiación ionizante. Uno de los objetivos de este trabajo ha sido desarrollar una técnica de artrografía de la articulación de la rodilla canina, determinando la combinación de concentración y volumen de contraste, así como la efectividad de la artrografía para delimitar las superficies articulares y ligamentos de la articulación de la rodilla. Por otro lado hemos querido describir la anatomía ecográfica y por IRM normal de la rodilla canina y correlacionar de forma directa los cortes anatómicos con las imágenes ecográficas y de RM, para facilitar su interpretación. Para determinar la concentración y volumen de contraste del estudio astrográfico se realizó una experiencia previa en 3 animales en los que se combinaron diferentes volúmenes y concentraciones de iohexol, contraste iodado hidrosoluble no iónico de baja osmolaridad. La combinación elegida fue la de 0'4ml de iohexol por centímetro de grosor mediolateral de la articulación a una concentración de 300mgI/ml. El estudio artrográfico se llevó a cabo en 12 animales de tamaño mediano y grande, y los disparos se realizaron a los tiempos O, 3, 5, 10, 15, 20 Y 30 minutos tras la administración del contraste en proyecciones lateral y caudocraneal De los 12 artrogramas, 11 (91'6%) fueron clasificados como aceptables, resultando diagnósticos hasta el tiempo 10 en la proyección caudocraneal (CdCr) y hasta el 15 en la mediolateral (ML), posteriormente la absorción del contrastese incrementaba no pudiéndose discernir las estructuras articulares. El ligamento cruzado craneal, el receso suprarrotuliano y el cartílago articular de los cóndilos del fémur se clasificaron como obvios, claramente delineados y accesibles para la evaluación radiográfica, mientras que el resto de estructuras articulares fueron visualizadas y resultaron distinguibles y eventualmente accesibles para la evaluación radiográfica. No se encontraron diferencias significativas (p< 0'05) entre los dos grupos de animales, In dogs, the stifle is the most frequently injured joint, ligamentous and meniscal injuries being the most common, with its associated secondary degenerative changes. Diagnosis of stifle joint disorders is generally based on history of lameness, physical examination and radiography. Diagnostic imaging techniques Iike arthrography, ulttasonography and magnetic resonance imaging could be used to further evaluate these sttuctures. Knee arthrography is a cheap technique easy to perform, meanwhile all other are more sofisticated and expensive methods. Arthrographic studies have been used along the years in human beings, specially in the diagnosis of meniscal and cruciate ligaments injuries. Canine arthrography of this joint has received little attention in the veterinary literature. Ulttasound may probe to be useful in the diagnosis of cartilage abnormalities, meniscal tears, muscle, tendon and ligament abnormalities, arthropathies and neoplasia. The ability of ulttasound to image intra-articular soft tissues, allows it to provide additional information that can be used in conjunction with radiography. Magnetic resonance imaging (MRI) is a valuable diagnostic tool in humans, but is relatively new to veterinary medicine because is an expensive method and few veterinary centers currently have the equipment avaliable. MRl has become the preferred imaging modality for the evaluation of the articular cartiIage, menisci and ligaments of synovial joints. Unlike arthrography, MRl is noninterventional and does not require the use of ionizing radiation. The present study was done to evaluate the technique of stifle arthrography in the dog, to determine the volume and concentration of the contrast material and to determine the accurate evaluation of intraarticular structures and articular surfaces. We also wanted to describe the normal ultrasonographic appearance as well as the normal anatomy by MRl To conlude, we can say that arthrography performed with iohexol, does not provide a full anatomic detail of all structures of the stifle joint in dogs, although some articular structures as we said before could be perfectly accesible for radiographic evaluation. Is a gafe technique because no inflamatory changes in the synovial fluid 7 days following injection of iohexol was observed. Ultrasonography of the canine stifle could be a useful clinical application to noninvasively evaluate most of the intraarticular structures, meanwhile the increased soft tissue contrast resolution with MRI provides better detail of the soft tissue structures of the knee than conventional radiography and ultrasound and therefore is the best diagnostic imaging tool to evaluate injuries at the canine stifle joint. arthrographic procedures were clear, colorless and with citologic values remained within normal limits in all dogs.

La hipòtesi establerta va ser que la modificació oxidativa de les lipoproteïnes de baixa densitat (LDL) i de les lipoproteïnes d'alta densitat (HDL) augmenta el potencial aterogènic d'aquestes lipoproteïnes. Estudis anteriors havien mostrat que les LDL oxidades eren reconegudes pel receptor depurador dels macròfags i afavorien l'acúmul de colesterol dins de la cèl·lula, associat a la transformació del macròfag en cèl·lula espumosa, punt central en la formació de la placa d'ateroma. Recentment, s'ha observat que la modificació oxidativa de les HDL provoca una disminució de la sortida del colesterol cel·lular i afavoreix un major acúmul de colesterol dins de la cèl·lula. Per tant, la modificació oxidativa de les LDL i de les HDL pot jugar un paper important en la formació i desenvolupament de l'arteriosclerosi.Per un altre costat, sabem que la dieta modula la composició i el metabolisme de les LDL i HDL, la qual cosa explica, almenys en part, la diferent aterogenicitat lligada a les dietes. En particular, el nostre interès es va centrar en el fet que l'enriquiment en àcid oleic de les partícules lipoproteiques, a través de la dieta, proporcionava una protecció addicional enfront dels fenòmens oxidatius.Vam estudiar les LDL i HDL com a principals partícules lipoproteiques implicades en el transport lipídic i en la formació de la placa d'ateroma. Dins de les HDL vam estudiar les HDL3 per ser la subfracció més directament implicada en la sortida del colesterol de les cèl·lules.En el nostre estudi vam comparar els efectes d'una dieta rica en àcid linoleic, aportat per l'oli de gira-sol, i d'una dieta rica en àcid oleic, subministrat per un oli de gira-sol modificat per tal d'enriquir-lo en aquest àcid, sobre:-Les concentracions en lípids, lipoproteïnes i apolipoproteïnes del plasma.-La composició química global de les fraccions LDL i HDL3.-La composició en àcids grassos dels fosfolípids de les HDL3.-La fluidesa de les LDL i HDL3.-La peroxidació lipídica de les HDL3 natives, així com la susceptibilitat a l'oxidació de les LDL i HDL3.-La concentració en plasma, LDL i HDL3 dels antioxidants biològics: -tocoferol i +-tocoferol i retinol.-La capacitat de les LDL aïllades després de les dues dietes per a ésser reconegudes pel receptor depurador dels macròfags humans.-La capacitat de les HDL3 aïllades després de les dues dietes per extreure el colesterol cel·lular en fibroblasts i macròfags humans.Les LDL i HDL van ser obtingudes del plasma de 22 monjos sans amb una edat (mitjana  ES) de 53.7  3.1 anys. Aquests individus van ser assignats a l'atzar a dos grups de 11 individus cadascun d'ells. Es va calcular una dieta isocalòrica amb un aport de greixos del 38%, amb un 15% del total calòric provinent de l'oli (oli de gira-sol convencional o bé una nova varietat d'oli de gira-sol que conté més d'un 80% d'àcid oleic). Els nostres resultats reafirmen la hipòtesi que una dieta rica en àcid oleic augmenta la resistència de les LDL i de les HDL3 a l'estrès oxidatiu, a igualtat en el contingut d'antioxidants. Aquest fet explica, almenys en part, la menor aterogenicitat lligada a una dieta rica en àcid oleic. Per un altre costat, aquestes dues dietes, l'una rica en àcid oleic i l'altra rica en àcid linoleic, que provenen d'olis diferents originats a partir de la llavor de gira-sol, indueixen unes lipoproteïnes que han mostrat, segons les característiques estudiades, una estructura i un comportament metabòlic similar.Aquests resultats confirmen els efectes de la dieta en l'oxidació i contribueixen a explicar els mecanismes d'acció dels àcids grassos de la dieta en l'arteriosclerosi., This study was conducted to evaluate whether diets rich in oleic acid could result in HDL3 particles with greater resistance to oxidative modification compared with a linoleic acid enriched diet, as it has been observed for LDL.Plasma LDL and HDL3 were obtained from 22 healthy monks aged 53.7  3.1 years (mean  SE), who were randomly assigned to a 32-wk crossover study of a variant of sunflower oil containing more than 80% oleic acid (oleic acid diet) compared with conventional sunflower oil containing 67% linoleic acid (linoleic acid diet). The two diet test periods of 8 weeks each contained 38% of total calories as fat respectively. HDL3 isolated after the oleic acid diet were enriched in oleic acid in phospholipids. LDL and HDL3 obtained after the oleic acid diet were remarkably resistant to oxidative modification. Plasma lipids, lipoproteins and apolipoprotein (apo) B concentrations showed no differences after the two diets, but higher levels of apo A-I and A-II were observed after the oleic acid diet. LDL and HDL3 composition, fluidity and vitamins E, A and + tocopherol concentrations were not significantly different between the two diets. Competition studies with 125I-Ox LDL in macrophages were similar after the two diets. The capacity of HDL3 to induce [3H]free cholesterol efflux from fibroblasts and macrophages also showed no differences after the two diets.These results suggest that, like LDL, HDL3 rich in oleic acid are protected against oxidation, the resistance of these lipoproteins to oxidation being mainly determined by their oleic acid content This study demonstrates the feasibility of a diet rich in oleic acid as an attractive way to cardiovascular dietary prevention.

Es conocido que las sinapsis cambian en condiciones de normalidad durante el desarrollo que culmina en la maduración. Sin embargo, se han identificado cambios funcionales de corta duración actividad-dependientes en adultos normales. Así en el decremento de uso aparecen fenómenos de crecimiento axonal por elongación de algunos segmentos axonales. Y en el incremento de uso, dominan los fenómenos de retracción de algunas ramas axonales. En este contexto, el presente trabajo tiene como pretensión final conocer de forma clara la capacidad de adaptación plástica de las conexiones neuromusculares en el adulto normal.MATERIAL Y METODOS: Se utilizó el músculo Extensor digitorum longus (EDL) de ratas Sprague-Dawley macho, de 12 semanas de edad. Los animales fueron sometidos a un moderado incremento de actividad locomotora mediante una cinta sin fin. Las bandas de inervación de los músculos EDL fueron procesadas para microscopia electrónica de transmisión convencional. Con las imágenes obtenidas (a 5000x y 10000x) se realizo un estudio morfométrico con el programa semiautomático de análisis de imágenes MOP VIDEOPLAN.CONCLUSIONES MÁS RELEVANTES:1- La retracción axonal en músculos control es compensado por los siguientes cambios en estructuras relacionadas con la neurotransmisión: mas y mayores vesículas sinápticas y también incrementa el numero de vesículas que están en contacto con el axolema sináptico; mas zonas activas axonales; las membranas densificadas de las crestas postsinápticas decrecen en longitud pero esto es compensado por un mayor número.2- Los resultados morfométricos indican claramente que pequeños cambios en el nivel de actividad locomotora inducen a precisas adaptaciones morfológicas de una forma rápida. El incremento en las densificaciones postsinápticas sugiere un mecanismo compensatorio postsináptico a la retracción axonal., The activity pattern of neuromuscular junctions can influence the structure, growth and remodeling responses and differentiation of the motor nerve terminals. Nerve terminal remodeling involving changes of growth and retraction in the normal adult are affected by the level of the animal's activity. In fact, nerve terminal retraction in the adult can be induced by the level of increased synaptic activity. In the present work we tried to analyze if a small physiologic increase in the locomotor's activity of the normal adult rats during a brief period of time can induce pre- and postsynaptic ultrastructural adaptations.MATERIAL AND METHODS: The Extensor Digitorum Longus (E.D.L.) muscles of 24 three-month-old male Sprague-Dawley rats were studied. The experimental animals spontaneously adapted to the moderately exercise by training a treadmill. Normally housed rats were used as controls. The neural zones from the right and left E.D.L. muscles were fixed in glutaraldehyde and conventionally processed for T.E.M. Cross-sectioned different end plates were directly recorded in magnetic support with a high definition video system. Morphometric analysis of pre- and postsynaptic structural parametres was performed with a semiautomatic image analysis system (MOP VIDEOPLAN).MAIN CONCLUSIONS: 1- Axonal retraction in normal muscles can be compensated by the following changes in the neurotransmission-related structures: Synaptic vesicles increase in diameter and also increase the number of synaptic vesicles in contact with the synaptic axolemma; active zones increase in number; postsynaptic membrane densifications decrease in length and that is compensated by an increase in number of these.2- The morphometric data clearly indicate that small and early physiological changes in the locomotor's activity in the normal adult are followed by precise morphologic adaptations. The increased differentiation of postsynaptic membrane densifications would suggest the existence of the postsynaptic compensatory mechanism for the presynaptic retraction.

Young gerbil livers and kidneys were analyzed by means of light and electron microscope to assess the histopathological changes caused by prolonged systemic aluminum (Al) administration. The experimental group was injected with AlCl3 i.p. for 5 weeks, while litter mates received PBS as sham-injected controls or served as untouched controls. Mortality occurred in 33% of experimental and 12.5% of sham-injected groups. The animals were perfused intracardially with 1% glutaraldehyde plus 1% paraformaldehyde and samples of liver and kidneys were processed for aluminum and iron histochemistry and conventional light- and transmission electron microscopy. White deposits composed of cellular debris appeared on the surface of liver and kidneys and in the mesentery as a consequence of Al treatment. Adherences of Glisson capsule to the diaphragm, as well as scattered small foci of hepatocyte necrosis with non-caseificant microgranulomas and mild portal inflammation, developed in the experimental group. Sham-injected animals also exhibited these granulomas but to a lesser degree. Al deposits were found in experimental animal granulomas and inside macrophages cytoplasm scattered throughout the liver. Iron deposition appeared in pericentral hepatocytes of experimental animals, in granulomas and in portal spaces of the three groups of animals. Ultrastructurally, hepatocytes of experimental animals showed mitochondria hyalinization, disintegration of endoplasmic reticulum and clustering of ribosomes. Phagolysosomes appeared larger and occurred more frequently in both hepatocytes and Kupffer cells of experimental animals. In 2 out of the 6 experimental animals studied, tubular atrophy was present in the renal cortical region, the kidneys of the remaining animals appearing normal. Al and iron were found very occasionally in the kidney parenchyma of experimental animals, while isolated mesangial cells showed iron deposits in a few glomeruli of both experimental and the two groups of control animals.

The Hodgkin lymphoma (HL) is a B-cell lymphoma, as was proved by molecular studies with single-cell PCR. Histologically, it is characterized by a minority of neoplastic cells, Reed-Sternberg cells and its variants, related to a variable non-neoplastic inflammatory background. Nowadays, (WHO classification) the following types of HL are recognized: Nodular Paragranuloma and the Classical Hodgkin Lymphoma, the latter including Nodular Sclerosis, Mixed Cellularity, Lymphocyte-rich Classical Hodgkin Lymphoma and Lymphocyte Depletion. Morphology together with immunohistochemical studies allows to classify the different forms of Hodgkin lymphoma and to make a differential diagnosis with non-Hodgkin lymphomas. All classical Hodgkin lymphomas are treated similarly, and chances for remission and survival are currently good. Molecular parameters should be added to the current classification and patients could benefit from new therapeutic targets.

Gastric cancer is the second leading cause of cancer related death worldwide. In the UAE, recent data show an increase in the number of patients with gastric cancer highlighting the need for greater understanding of its pathogenesis. Gastric cancer is generally believed to develop on a background of chronic atrophic gastritis which eventually leads to intestinal metaplasia, dysplasia and finally invasive carcinoma. Recently this multistep process of carcinogenesis has been challenged. Therefore, the aim of this study is to define alterations in antral mucosal biopsies and cancer tissues to investigate whether they could be used to assemble a tissue array supporting the multistep model of carcinogenesis. Gastric mucosal tissues were obtained from informed individuals undergoing endoscopy (for upper gastrointestinal symptoms) and gastrectomy (for adenocarcinoma) in Tawam Hospital. All tissues were processed for microscopic examination. Eighty nine antral biopsies were categorized as: normal (33%), mild superficial gastritis (34%) and severe atrophic gastritis (33%). About 5% of the latter exhibited evidence of intestinal metaplasia. Cancer tissues obtained from three patients were microscopically examined in three regions: safe resected margin, tumor edge and tumor center. Progressive changes in mucosal thickness, dysplasia and cellular transformation were observed, and when compared with alterations in biopsies, all appeared to represent a continuum of progression toward invasive adenocarcinoma. In conclusion, the tissue array presented in this study supports the multistep process of gastric carcinogenesis and will be helpful in examining the expression pattern of tumor markers or molecules that could help in the early detection of gastric cancer.

Summary. Desmoid tumors are monoclonal proliferations that fall within a broad histologic spectrum of fibrous mesenchymal tumors that ranges from benign proliferations of scar tissue to high-grade fibrosarcomas. These low-grade tumors are extremely infiltrative locally, but lack the ability to metastasize systemically. While they are only rarely a direct cause of mortality, using current therapeutic modalities, these tumors have a high rate of local recurrence that can result in significant treatment related morbidity. Sporadic desmoids are usually associated with somatic mutations in codons 41 or 45 of exon 3 of ß-catenin (CTNNB1). Desmoid tumors occurring in the background of familial adenomatous polyposis (FAP) usually contain inactivating germline mutations in the adenomatous polyposis coli (APC) gene. CTNNB1 and APC are part of the Wnt signaling pathway and mutations in either gene result in stabilization of the ß-catenin protein and allow nuclear translocation and binding of ß-catenin to the T-cell factor/lymphoid enhancer factor (TCF/Lef) family of transcription factors, resulting in activation of target genes which may underlie desmoid tumor biology and clinical behavior. In an era of molecularly targeted therapeutics there is a real need to better grasp the molecular mechanisms behind desmoid tumorigenesis and progression. This knowledge will eventually result in the development of patient and tumor tailored therapies and assist in the control and eradication of this disease.

Temporomandibular joint internal derangement (TMJ ID) is characterised by disc displacement and degenerative tissue changes involving an active cellular response, with cell phenotype transformation from fibroblast-like to fibrochondrocyte and, eventually, to chondrocyte-like, possibly as a response to abnormal loading. However, only small patches of chondral tissue are detected in TMJ discs with ID. We decided to explore the reasons for such incomplete tissue change, postulating an involvement of the apoptosis process. Twenty-one discs removed from 19 patients with TMJ ID were processed for TRAIL and DR5 immunohistochemical localisation, and subjected to the TUNEL assay. Overexpression of DR5 receptor and its ligand (TRAIL) in chondrocyte-like cells suggested activation of programmed cell death, as also demonstrated by TUNEL-positive cells. The data suggest a failed adaptive response to disc displacement through chondroid metaplasia. The apoptotic death of chondrocyte-like cells, which is at least partly regulated by TRAIL and its death receptor, appears to underpin the failed disc repair, eventually leading to its perforation.

Trastuzumab has substantially changed the prognosis of breast carcinomas. As HER2 overexpression/ amplification is a prerequisite for treatment with trastuzumab, an accurate assessment of HER-2 status is the first step for successful treatment. In metastatic breast cancer, we routinely assess HER2 expression in the primary tumour, assuming that HER2 status remains stable through cancer progression. However, it is frequent to find reports that describe discordance between HER2 expression in primary and metastatic tumours. The aim of this paper was to verify whether HER2 status of breast carcinomas is maintained in the corresponding axillary metastasis. Immunohistochemistry was performed on 52 breast carcinomas and their matched axillary metastasis. HercepTest results were concordant in 46 out of 52 cases (88.5%). FISH proved that the differences observed were clinically relevant in only one of the 52 cases studied (98% concordance). We concluded that HER2 status was stable during axillary metastatic progression. Evaluation of gene HER2 status in axillary metastasis rather than in the primary can be useful in certain situations, e.g., small invasive component intimately mixed with in situ component and difficult to recognize in dark field, no tumor after biopsy, or axillary relapse (in this case we can find occasional de novo amplifications susceptible to trastuzumab treatment).

Claudins are tight junction proteins that are critical for the sealing of cellular sheets and controlling paracellular ion flux. The claudin family of proteins is composed of at least 24 closely related transmembrane proteins, most of them are well characterized at the gene and protein levels. The claudins are present in variety of normal tissues, hyperplastic conditions, benign neoplasms, and cancers that exhibit epithelial differentiation. Loss of claudins expression has also been reported in several malignancies as well. Differential expression of various members of the claudins family in cancers can be used in confirming the histologic identity of certain cancers and excluding others. Examples include the use of immunohistochemical detection of claudins to differentiate between oncocytoma and chromophobe renal cell carcinoma, endometrial endometrioid carcinoma and seropapillary carcinoma, mesothelioma and metastatic adenocarcinoma, hepatocellular and biliary tract carcinomas, and between intestinal-type and diffuse-type gastric carcinoma. Expression of certain claudins can also be used as markers that can predict patient’s prognosis. Thus, it seems that attempts to identify expression claudins in cancers are becoming increasingly useful in histologic diagnosis of tumors as well as means to assess patient’s prognosis.

Gut epithelial apoptosis is involved in the pathophysiology of multiple diseases. This study characterized intestinal apoptosis in three mechanistically distinct injuries with different kinetics of cell death. FVB/N mice were subjected to gamma radiation, Pseudomonas aeruginosa pneumonia or injection of monoclonal anti-CD3 antibody and sacrificed 4, 12, or 24 hours post-injury (n=10/time point). Apoptosis was quantified in the jejunum by hematoxylin and eosin (H&E), active caspase-3, terminal deoxynucleotidyl transferase dUTP-mediated nick end labeling (TUNEL), in situ oligoligation reaction (ISOL,) cytokeratin 18, and annexin V staining. Reproducible results were obtained only for H&E, active caspase-3, TUNEL and ISOL, which were quantified and compared against each other for each injury at each time point. Kinetics of injury were different with early apoptosis highest following radiation, late apoptosis highest following anti CD3, and more consistent levels following pneumonia. ISOL was the most consistent stain and was always statistically indistinguishable from at least 2 stains. In contrast, active caspase-3 demonstrated lower levels of apoptosis, while the TUNEL assay had higher levels of apoptosis in the most severely injured intestine regardless of mechanism of injury. H&E was a statistical outlier more commonly mechanism or kinetics of injury, ISOL correlates to other quantification methods of detecting gut epithelial apoptosis more than any other method studied and compares favorably to other commonly accepted techniques of quantifying apoptosis in a large intestinal cross sectional by balancing sensitivity and specificity across a range of times and levels of death.

Cervical displasia are classified as CIN-I, CIN-II and CIN-III. It has been observed that in at least 60% of CIN-I and CIN-II, the pathology disappears spontaneously, while around 30% persist at 24 months, 10% progress to CIN-III and 1% develops as a SCC. The factors involved in the evolution of the pathology are not defined, although infection of HPV is a necessary condition, but not the only one. For this reason, the identification of genetic changes is an essential element for understanding the carcinogenic process. It can also serve as a helpful tool for identifying patients who may be susceptible to its evolution and treatment, from patients whose lesions could regress spontaneous and for whom periodic follow-ups would be enough. Fiftty three cervical biopsies from patients with dysplasia and ISCC were included in the study. These biopsies were set into nine macroarrays. Eight genes and five proteins were examined in each samples (hTERT, PIK3CA, hTERC, MYC, CCND1, BCL2, ZNF217 and p16) by fluorescence in situ hybridization (FISH) and/or immunohistochemistry (IHC). The results reflected that the genetic alterations of PIK3CA, ZNF217 and CCND1 were associated with the evolution of normal tissue to CIN I, those of hTERC and ERBB with the evolution of LSIL to HSIL, those of hTERT and MYC with the evolution of CIN-II/CIN-III to ISCC, and those of BCL-2 with the inception of ISCC. With regards to proteins, the expression of MYC and CCND1 in the initial stages of the illness would help in the acquisition of the altered cellular phenotype.

Many investigators have undertaken radiologic studies in chelonians. However, descriptive papers focusing on the radiographic anatomy are limited to only a few species. The purpose of this article is to provide the normal cervical and coelomic radiographic appearance of the loggerhead sea turtle (Caretta caretta), in the dorsoventral view, and to indicate useful landmarks to identify internal anatomic structures. Dorsoventral radiographs were taken of the neck and body of 30 loggerhead sea turtles by means of analog and digital radiography. At various points, distortion or superimposition of images due to the natural curvature of the shell hindered the accuracy of interpretation. The pectoral and pelvic girdles were easily recognized. Important external landmarks included the vertebral and lateral scutes, and important internal landmarks included the bronchi, coracoid bones, the caudal border of the pulmonary fields, and the acetabulum.

Small GTPase Rabs constitute the largest family of membrane trafficking proteins that are conserved in all eukaryotic cells. The number of different Rab isoforms in multicellular organisms is usually greater than that in unicellular organisms (e.g., approximately 60 different Rabs in each species of mammals investigated versus approximately 10 Rabs in yeasts). The expansion of Rab isoforms in mammals is often regarded as due to the acquisition of specialized membrane trafficking events in the specialized cell types of higher eukaryotes. However, because of their large numbers the precise function of most mammalian Rab isoforms is still unknown. The recent development of new tools for comprehensive analysis (e.g., Rab panels) has paved the way for systematic investigation of the involvement of mammalian Rab isoforms in specialized membrane trafficking events. The tools include collections of enhanced green fluorescent protein (EGFP)-tagged mouse and human Rabs, FLAG-tagged Rabs, glutathione S-transferase (GST)-tagged Rabs, Gal4-binding domain (GBD)-tagged Rabs, Tre- 2/Bub2/Cdc16 (TBC) domain-containing Rab-GTPaseactivating proteins (GAPs), and small interfering RNAs. EGFP-Rabs are used to screen for Rabs that are localized on specific organelles and regulate their transport, and GST-Rabs and GBD-Rabs are used to screen for novel Rab effectors by GST pull-down assays and yeast two-hybrid assays, respectively. Combined use of these tools now makes it possible to efficiently determine the function of mammalian Rab isoforms in membrane traffic. This article reviews the development of new tools for systematic analysis of Rab proteins and their application to Rab-mediated membrane traffic.

We thank Dr. Vitor Carneiro (pathologist from the Hospital do Divino Espírito Santo, Ponta Delgada, São Miguel, Azores), for his help in retrieving the breast cancer samples for this study., The goal of the present work was to evaluate the correlation of glucose transporter 1 (GLUT1) and carbonic anhydrase IX (CAIX) with the monocarboxylate transporters 1 (MCT1) and 4 (MCT4) and their chaperone, CD147, in breast cancer. The clinico-pathological value of GLUT1 and CAIX was also evaluated. For that, we analysed the immunohistochemical expression of GLUT1 and CAIX, in a large series of invasive breast carcinoma samples (n=124), previously characterized for MCT1, MCT4 and CD147 expression. GLUT1 expression was found in 46% of the cases (57/124), while CAIX was found in 18% of the cases (22/122). Importantly, both MCT1 and CD147, but not MCT4, were associated with GLUT1 and CAIX expression. Also, GLUT1 and CAIX correlated with each other. Concerning the clinico-pathological values, GLUT1 was associated with high grade tumours, basal-like subtype, absence of progesterone receptor, presence of vimentin and high proliferative index as measured by Ki-67. Additionally, CAIX was associated with large tumour size, high histological grade, basal-like subtype, absence of estrogen and progesterone receptors and presence of basal cytokeratins and vimentin expression. Finally, patients with CAIX positive tumours had a significantly shorter disease-free survival. The association between MCT1 and both GLUT1 and CAIX may result from hypoxia-mediated metabolic adaptations, which confer a glycolytic, acid-resistant and more aggressive phenotype to cancer cells., CP and AA received fellowships from the Portuguese Foundation for Science and Technology (FCT), refs. SFRH/BD/27465/2006 and SFRH/BD/15316/2005, respectively. This work was supported by the FCT grant ref. PTDC/SAU-FCF/104347/2008, under the scope of “Programa Operacional Temático Factores de Competitividade” (COMPETE) of “Quadro Comunitário de Apoio III” and co-financed by Fundo Comunitário Europeu FEDER.

Loss of heterozygosity and chromosomal rearrangement of the WWOX gene, which is located at 16q23.3-24.1, have been detected in ovarian, breast, hepatocellular, and prostate carcinomas and in other neoplasias. This gene, which spans the common chromosomal fragile site 16D, contains 9 exons and encodes a 46 kDa WWOX protein that contains 414 amino acids. The evidence from cancer cell lines and primary tumor tissues suggests that WWOX is a tumor suppressor gene and that its inactivation contributes to cancer development. The results from studies of WWOX gene knockout cancer cells and a WWOX knockout mouse model partly confirm this hypothesis. The nature of the various proteins that the WWOX protein can interact with, such as c-Jun, TNF, p53, p73, AP- 2gamma, and E2F-1, suggests that WWOX plays a central role in tumor suppression through transcriptional repression and apoptosis, with its apoptotic function the more prominent of the two. However, there is not universal agreement that WWOX is a tumor suppressor gene. Further analysis is needed to reveal the true nature of WWOX.

We aimed to test whether antibodies raised against recombinant peptides corresponding to the variable region of immunoglobulin light chains are suitable for the immunohistochemical classification of amyloid. The Entrez database of the National Center for Biotechnology Information (NCBI) was searched for all protein sequence entries which met the search criteria “amyloid” and “lambda light chain”. Sixty-four different l-light chain-derived amyloid protein sequences were retrieved, aligned and categorized into the V region subgroups of l-light chain detailed by the NCBI, i.e. subgroup I (21 protein sequences), II (14), III (6), IV (1), V (1) and VI (21). V region subgroup I was chosen for epitope sequence selection and two rabbits were immunized with the following peptides: NH2- ISCSGSSSNIGSNTV-CONH2 and NH2-QRPSG VPDRFSGSKSGTS-CONH2. Sensitivity and specificity of the IgG-purified antibodies was tested by Western blotting using amyloid A- (AA), ALl- and ALk-amyloid proteins, and by immunohistochemistry on tissue microarrays with 110 different amyloid containing tissue samples obtained at autopsy from 22 patients, and on 27 biopsy specimens from a series of 24 patients. Our peptide antibodies specifically stained AL amyloid l- light chain-origin, in both Western blots and formalinthat peptide-antibodies directed against immunoglobulin- derived l-light chain proteins can be applied for the immunohistochemical classification of amyloid. This offers the opportunity to generate a large set of anti- l-light chain protein-antibodies for the immunohistochemical classification of amyloid independently from native human tissue sources.

An association between rheumatoid arthritis (RA) and malignancies has been ascertained and patients with RA appear to be at higher risk of lymphoma and lung cancer. The higher risk of the latter malignancy may be related to rheumatoid interstitial lung disease and immunosuppressive therapies. Herein we illustrate the case of a 59-year-old male smoker affected by RA and treated with cortisone, methotrexate and TNF-α antagonists, who underwent right lower lobectomy for a nodular lesion. On microscopic examination, the lesion consisted of two distinct areas: a central area of fibrinoid necrosis, bordered by histiocytes in a palisaded arrangement, lymphocytes and a 0.4 cm thick peripheral area constituted by a combined small cell anaplastic carcinoma, adenocarcinoma and squamous cell carcinoma. The combination of three histotypes is very rare in such a small tumour. In our case, it may be hypothesized that synchronous, heterogeneous mutations occurred in different type of committed cells or in stem cells, due to the production of cytokines by RA nodule histiocytes and lymphocytes, which are contiguous to the carcinomatous area. Since few studies have evaluated the topographic correlation between tumors and rheumatoid lung lesions, further morphological and molecular studies are needed to clarify this association and the pathogenetic relationship between RA and cancer of the lung.

Recent data challenge the relevance of the RB pathway to cancer based on RB inactivation, at least in breast tumors. To obtain information on the actual role of the components of the RB pathway in tumor progression we decided to investigate whether their quantitative changes were associated with variations in the level of RB phosphorylation in human breast cancer. A series of 68 human primary breast carcinomas was studied. Five cases were excluded from the study due to their lack of RB expression. In the remaining 63 cases the expression of cyclin D1, cdk4, cyclin E, and INK4a mRNA was assessed by real-time RT-PCR. The level of RB phosphorylated protein (ppRB) and p27 expression was immunohistochemically analyzed by measuring the percentage of stained cells (labeling index, LI). Cell proliferation rate was measured by Ki67 LI evaluation. The ppRB LI ranged from 5.2 to 73.8 and, as expected, was strongly related to the Ki67 LI (r=0.80; p

Summary. Aims: The purpose of this study was to draw up a TMJ disc histopathological score that is a semiquantitative transcription of the entire spectrum of TMJ disc degenerative diseases related to changes in disc tissue, and then validate the proposed grading, in order to contribute to a standardized histopathological diagnosis. Methods: Sections from sixty two temporomandibular joint disc specimens affected by tissue degenerative changes and stained with Hematoxylin & Eosin (H&E) were collected from among those found in the archives of the Department of Dentistry at Catania University. Specimens, included anterior, intermediate and posterior disc bands. Based on a literature search regarding the most frequent histopathological changes detected in TMJ disc tissue a grading score was designed. This score takes into account pathological disc tissue transformation i.e. collagen bundles, non-specific degenerative changes and the presence of blood vessels. This grading system results in a score ranging from 0 up to 8 for heavily degenerated disc tissue. Two observers performed the assessment of the TMJ disc H&E stained sections. Each specimen was scored twice by each observer after a minimum interval of 1 week. Results: The average TMJ disc degeneration score of the TMJ disc sample was 3.89±1.37. There was an almost perfect agreement, between the two observers, and degeneration scores from the two observers were highly correlated. Conclusions: The introduction of this validated degeneration score system may be of major importance for future research and collaboration between different centers in order to improve knowledge in TMJ disc histopathology.

Tumorigenesis in human glioblastoma multiforme (GBM) is driven by several genetic abnormalities with disruption of important molecular pathways, such as p53/MDM2/p14ARF and EGFR/PTEN/Akt/mTOR. The malignant progression of human GBM is also primarily associated with a peculiar multistep pathophysiological process characterized by intratumoral ischemic necrosis (i.e. pseudopalisading necrosis) and activation of the hypoxia-inducible factor (HIF)-1a pathway with consequent peritumoral microvascular proliferation and infiltrative behaviour. Predictable preclinical animal models of GBM should recapitulate the main pathobiological hallmarks of the human disease. In this study we describe two murine orthotopic xenograft models using U87MG and U251 human cell lines. Ten Balb/c nude male mice were orthotopically implanted with either U87MG (5 mice) or U251 (5 mice) cell lines. Intracranial tumor growth was monitored through Magnetic Resonance Imaging (MRI). Immunohistopathological examination of the whole cranium was performed 30 days after implantation. U251 orthotopic xenografts recapitulated the salient pathobiological features described for human GBM, including invasive behaviour, wide areas of pseudopalisading necrosis, florid peripheral angiogenesis, GFAP and vimentin expression, nonfunctional p53 expression, striking active-caspase-3 and HIF-1a expression along pseudopalisades. U87MG orthotopic xenografts proved to be very dissimilar from human GBM, showing expansile growth, occasional necrotic foci without pseudopalisades, intratumoral lacunar pattern of angiogenesis, lack of GFAP expression, fuctional p53 expression and inconsistent HIF-1a expression. Expression of pAkt was upregulated in both models. The results obtained suggest that the U251 orthotopic model may be proposed as a predictive and reliable tool in preclinical studies since it recapitulates the most salient pathobiological features reported for human GBM.

The computed tomography (CT)-based early lung cancer diagnostic technologies allow the detection of very small stage I lung tumors. As part of these screening protocols any suspicious nodule has to be diagnosed morphologically, which requires CT-guided Fine Needle Aspiration, open biopsy or surgery. Fine Needle Aspiration (FNA) cytology is a well-recognised method for a rapid and accurate diagnosis of small lung tumors. Molecular analysis of the FNA specimens could complement cytology diagnosis by the characterization of the biological traits at the preoperative stage. In this study, we aimed to characterize the biological profile of 33 paraffin-embedded transthoracic FNA samples obtained from three groups of lung cancer patients: two groups of small early-detected lung adenocarcinomas (radiologically subsolid and solid nodules) and a third group of small metastatic adenocarcinomas. Genetic analysis was performed by fluorescence in situ hybridization using the four-color LAVysion probe. p53 and Ki-67 protein expression was also evaluated by immunocytochemistry. The samples showed gains for all targets analyzed; two cases had EGFR gene amplification and two cases had MYC amplification. There were no significant differences in the percentage of genetically malignant cells and the expression of Ki- 67 among the three groups. However, p53 accumulation was significantly higher in the metastatic group compared to the subsolid early-detected group (P = 0.001). In conclusion, molecular analysis of FNA specimens may provide useful information at preoperative stages. In our series, a good prognostic profile in subsolid early detected adenocarcinomas is suggested.

Podoplanin is involved in tumorigenesis and cancer progression in head and neck malignancies and its expression is not restricted to lymphatic vessel endothelium. The aim of this study was to establish podoplanin expression in the tumor-free resection margins of oral squamous cell carcinomas (OSCCs) and to evaluate the geometric complexity of the lymphatic vessels in oral mucosa by utilizing fractal analysis. As concerns the podoplanin expression in noncancerous tissue, forty tumor-free resection margins from OSCCs were investigated utilizing immunohistochemistry for D2-40 antibody and image densitometry analysis. Podoplanin expression was extremely low in basal cells, especially in resection margins of OSCCs developed in the lower lip regions. However, a highly variable D2-40 expression in tumor-free resection margins associated with hyperplastic or dysplastic lesions was identified. Moreover, podoplanin expression also extended to the basal layer of the lower lip skin appendages, the myoepithelial cells of acini and ducts of minor salivary glands, and other structures from the oral cavity. As concerns the study of the density and complexity of oral lymphatic vessels architecture by means of immunohistochemistry (D2-40, CD31 and Ki-67 antibodies) and fractal analysis, we demonstrated that in normal oral mucosa the geometry of the lymphatic vessels was less complex at the level of the lower lip compared to the anterior part of the oral floor mucosa or the tongue. A comparative analysis between the normal and pathological aspects revealed statistically significant differences between the fractal dimension (FD) of the vessels’ outline, especially in the tongue. Fractal analysis proved an increasing lymphatic network complexity from normal to premalignant oral mucosal lesions, providing additional prognostic information in oral malignant tumors

The aim of the study was to evaluate the performance of immunohistochemical MS110 expression in a series of familial and sporadic breast cancer patients. An immunohistochemical study was performed on TMA samples from 93 sporadic and 94 familial breast cancer patients with (7/94) and without BRCA1 germline mutations. BRCA1 protein expression level was evaluated using the monoclonal MS110 antibody. Immunohistochemistry, performed on TMA samples, showed positive nuclear staining for BRCA1 in 34 sporadic and 37 familial breast tumours, respectively. All the tumours from patients carrying BRCA1 mutations showed complete loss of both BRCA1 and ERa expression, regardless of the type of mutation. The percentage of MS110 positive cases was significantly lower in mutated versus wild type BRCA1 familial cases (p=0.02) while the percentage of patients with higher ERa expression was significantly lower in BRCA1- mutated versus BRCA1-wild type familial patients (p=0.05). Interestingly, the presence of the E1038G polymorphism in BRCA1 exon 11 was significantly associated with protein expression (p=0.029). The frequency of MS110 negative cases also detected in BRCA1-wild type tumours, points to the inability of the BRCA1 IHC expression in discriminating between familial and sporadic breast cancer.

EPH receptors and their ephrin ligands constitute the largest sub-family of receptor tyrosine kinases (RTKs) and are components of cell signaling pathways involved in animal development. The ability of the EPH/ephrin guidance system to position cells and modulate cell morphology underlies their various roles in development. In addition, EPH signaling plays an important role in oncogenic processes observed in several organs. These receptors are involved in a wide range of processes directly related with tumorigenesis and metastasis, including cell attachment and shape, migration, and angiogenesis. Accordingly, deregulation of EPH expression and signaling activity could be crucial for the tumorigenic process. This review focuses on EPH receptors’ roles in oncogenic transformation and tumor progression.

Granular cell tumor (GCT) is a relatively rare neoplasm, usually located in the upper aerodigestive tract, skin and soft tissue. Because of its uncertain histogenesis, GCT has been the object of many immunohistochemical and ultrastructural studies that have suggested a Schwann cell origin. Our recent observation of a case of GCT immunoreactive for Galectin-3 and HBME-1 led us to further investigate the immunohistochemical profile of these neoplasms. We evaluated the immunohistochemical expression of the traditional markers for GCT (S-100, CD68) along with new markers (Galectin-3, HBME-1, Calretinina and Inhibin-a) in 22 granular cell tumors. Our results showed, in all cases, a constant diffuse positivity for S- 100 protein, CD68 and Galectin-3. HBME-1 was positive in 95% of cases. The present study gives a new immunophenotypic profile for GCT, which could help pathologists in distinguishing morphologically ambiguous granular lesions in unusual sites.

Objective: Ghrelin is predominantly produced by neuroendocrine cells of stomach and has been expressed in several normal and tumour endocrine tissues. It has been reported that the localization of ghrelin is exclusively in the cortex of human and rat adrenal gland and in adrenocortical tumours. This prompted us to analyze the expression of this peptide in medulla of human and rat adrenal glands and in human pheochromocytomas. Design and methods: Analysis of ghrelin mRNA expression in rat adrenal gland was conducted by means of semi-quantitative RT-PCR. Ghrelin localization was studied in medulla of human and rat adrenal gland by immunohistochemistry. In addition, we have carried out a double immunofluorescence with chromogranin A to determine the specific cell type expressing ghrelin immunoreactivity. Ghrelin expression was also analyzed in five cases of pheochromocytoma by immunohistochemistry. Finally, Western blotting analysis was performed with goat ghrelin antibody in the cortex and in the medulla of rat adrenal gland. Results: RT-PCR demonstrated expression of ghrelin mRNA in rat adrenal gland. We also detected ghrelin expression in virtually all rat pheochromocytes by immunohistochemistry and double immunofluorescence. Furthermore, we showed ghrelin immunoreactivity in the medulla of human adrenal gland and in pheochromocytomas. By Western blotting, we found the expression of ghrelin precursor, proghrelin and mature ghrelin in the medulla of rat adrenals. However, the cortex of rat adrenal gland only expressed ghrelin precursor. Conclusions: Our study is the first to demonstrate a medullar expression of ghrelin in human and rat adrenal gland; we also showed ghrelin expression in pheochromocytomas.

Microvascular density (MVD) counting protocols have become the morphological gold standard to assess the neovasculature in human tumors. This method requires the use of specific markers to vascular endothelium and of immunohistochemical procedures to visualize microvessels. MVD determined in primary tumors is significantly associated with metastasis and prognosis in several tumors and is most predictive in those tumors that induce significant angiogenesis, namely carcinomas of breast and prostate, and haematological malignancies. There is such a wide range of antibodies and suppliers, antigen retrieval methods, designation of high and low vessel count groups, patient study groups and data interpretation, that it is exceedingly difficult to compare results.

Aims: The intake of salt and salty food is known as a risk factor for gastric cancer. We have previously demonstrated that a high-salt diet dosedependently enhances Helicobacter pylori (H. pylori)- associated gastritis and stomach carcinogenesis in Mongolian gerbils. In this study, we focused on the influence of excessive salt intake on the expression of inflammatory mediators involved in progression of H. pylori-induced chronic gastritis. Methods and Results: A total of 45 stomach samples from Mongolian gerbils were evaluated by immunohistochemistry. The animals were infected with H. pylori and fed basal (0.32%) or a high-salt (10%) diet, and sacrificed after 40 weeks. Proliferative activity and expression of cyclooxygenase-2 (COX-2) in gastric mucosa were significantly increased in H. pyloriinfected gerbils. The additional high-salt diet significantly up-regulated the expression of inducible nitric oxide synthase (iNOS) and COX-2 in H. pyloriinfected groups (P

In recent years, classification of soft-tissue sarcomas (STS) has improved with cytogenetic analyses, but their clinical behavior is still not easily predictable. The aim of this study was to detect alterations in the urokinase-type plasminogen system, involved in tumor growth and invasion, by comparing mRNA levels of its components with those of paired normal tissues, and relating them with patient clinical course. Real-time PCR was performed on human STS cell lines and tissues from highly malignant STS, including leiomyosarcomas and malignant fibrous histiocytomas, to evaluate the expression of urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1). Immunohistochemistry of gene products was also performed. Median mRNA values of all genes studied were higher in tumors than in paired normal tissues. In agreement with data on STS cell lines, significant upregulation for uPA and PAI-1 genes compared to reference values was seen. Moreover, different levels of expression were related to histotype and metastatic phenotype. There was accordance between uPA mRNA and protein expression, while immunodetection of PAI-1 product was weak and scattered. In recent years, classification of soft-tissue sarcomas (STS) has improved with cytogenetic analyses, but their clinical behavior is still not easily predictable. The aim of this study was to detect alterations in the urokinase-type plasminogen system, involved in tumor growth and invasion, by comparing mRNA levels of its components with those of paired normal tissues, and relating them with patient clinical course. Real-time PCR was performed on human STS cell lines and tissues from highly malignant STS, including leiomyosarcomas and malignant fibrous histiocytomas, to evaluate the expression of urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1). Immunohistochemistry of gene products was also performed. Median mRNA values of all genes studied were higher in tumors than in paired normal tissues. In agreement with data on STS cell lines, significant upregulation for uPA and PAI-1 genes compared to reference values was seen. Moreover, different levels of expression were related to histotype and metastatic phenotype. There was accordance between uPA mRNA and protein expression, while immunodetection of PAI-1 product was weak and scattered.

The stratified squamous epithelium of the nipple-areola complex may contain pale or clear cells including: Paget’s disease cells (PDCs), Toker cells (TCs), and so-called clear cells (CCs). Paget’s disease is an uncommon presentation of breast carcinoma. PDCs are large, atypical, have abundant, pale-staining cytoplasm that may contain mucin secretion vacuoles and bulky heterochromatic nuclei. They are commonly concentrated along the basal layer and stain for EMA, CAM5.2, cytokeratin 7, and HER2/neu oncoprotein. TCs are bland cells with roundish and scant chromatin nuclei. They are found incidentally and are reactive for EMA, CAM5.2, and cytokeratin 7, but show negativity for HER2/neu oncoprotein. So-called CCs show varied morphology, are found incidentally, and have been variably interpreted by different authors. The majority of cells that have been called epidermal CCs fit the features of pagetoid dyskeratosis. These cells are reactive for high molecular weight cytokeratin. Other CCs showing signet-ring morphology present negativity for mucins and correspond to a fixation artefact.

The aim of the present study was to ascertain the relationship between the level of RB1 mRNA and the expression of phosphorylated RB protein and the relevance of these two parameters in cancer cell proliferation and clinical outcome in human breast cancer. Sixty-eight primary human breast cancers were considered. The amount of RB1 mRNA was evaluated by quantitative RT-PCR analysis. The level of RB phosphorylation was immunohistochemical defined by measuring the phosphorylated (pp) RB labelling index (LI). Cell proliferation rate was measured by calculating the Ki67 LI. No relation was found between the RB1 mRNA level and the ppRB LI (p=0.565). Both RB1 mRNA value and ppRB LI were related (in an inverse and direct manner, respectively) to Ki67 LI. RB1 mRNA expression was more strictly associated with KI67 LI (p=0.001) than the ppRB LI (p=0.013). Regarding the patient clinical outcome, the separately considered RB parameters did not reach the prognostic significance. However, patients with low RB1 mRNA quantity and patients with high ppRB LI, taken together, had a significantly shorter disease free and overall survival than the group comprehending patients with high RB1 mRNA value and low ppRB LI, and this despite the low number of patients considered. Our results demonstrated that the ppRB LI was independent of the RB1 mRNA level; that both RB parameters are related to the cell proliferation rate and, if collectively considered, have a high informative value on breast tumour prognosis.

Angiogenesis is a dynamic, hypoxiastimulated and growth factor-dependent process, eventually leading to the formation of new vessels from pre-existing blood vessels. In the last decade experimental and clinical studies have described the occurrence of hepatic angiogenesis in a number of different pathophysiological conditions, including those involving inflammatory, fibrotic and ischemic features. In particular, the literature evidence indicates that hepatic angiogenesis is strictly associated with, and may even favour fibrogenic progression of chronic inflammatory liver diseases of different aetiology. In this review, current “in vivo” and “in vitro” evidence supporting the potential pathogenetic role of angiogenesis in chronic liver diseases will be reviewed in an attempt to outline cellular and molecular mechanisms involved, with a specific emphasis on the crucial role of hypoxic conditions and hepatic stellate cells (HSCs), particularly when activated to the myofibroblast-like pro-fibrogenic phenotype.

Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase that resides at the sites of integrin clustering, known as focal adhesions. The FAK protein has a molecular mass of 125kDa and is encoded by the FAK gene located on human chromosome 8q24. Structurally, FAK consists of an amino-terminal regulatory FERM domain, a central catalytic kinase domain, two proline-rich motifs, and a carboxy-terminal focal adhesion targeting domain. FAK has been shown to be an important mediator of cell growth, cell proliferation, cell survival and cell migration, all of which are often dysfunctional in cancer cells. Our lab was the first to isolate FAK from primary human tissue and link it to the process of tumorigenesis. We analyzed FAK mRNA expression in normal, invasive and metastatic human tissues and demonstrated through Northern blot analysis that normal tissues had very low levels of FAK mRNA while primary and metastatic tumors significantly overexpressed FAK. We also demonstrated and confirmed FAK overexpression in colorectal carcinoma and liver metastases with real-time PCR. In this review we summarized immunohistochemical data of FAK expression and role in different cancer types tumors and discussed FAK inhibition therapy approaches.

EGFR kinase activity triggers numerous signaling pathways, such as the Ras/Raf/MAPK cascade, leading to the activation of various mitogen activated protein kinases, which are implicated in cell proliferation through induction of several genes, including c-fos. The possible effect of diabetes on the expression of the oncogenes EGFR, H-ras and c-fos was investigated in an experimental model of chemically induced oral oncogenesis in normal and diabetic (type I) Sprague- Dawley rats. Thirteen diabetic and twelve normal rats developed cancer after 4NQO treatment, while six diabetic and six normal animals were used as controls. The biopsies were classified pathologically (ranging from dysplasia to moderately differentiated oral squamous cell carcinoma) and were studied immunohistochemically. Several representative histological regions from each biopsy were analysed in regard to EGFR, H-ras and c-fos expression, and a comparison between normal and diabetic rats was effected. A trend of decreased EGFR expression in diabetic compared to normal rats was revealed throughout oncogenesis, which was significant in the stage of dysplasia (P

At an ISUP (International Society of Urological Pathology) consensus conference in 2005 in San Antonio, Texas, the old Gleason grading system for prostatic carcinoma from 1966 underwent its first major revision. With this modified Grading system a shift of the most frequent Gleason scores from 6 to 7a (3+4) in biopsy specimens and an increased degree of agreement between specimens of biopsies and radical prostatectomies with carcinoma of the prostate could be demonstrated. After modified grading of GS 3+4=7a tumours 95% were stage pT2, while 79% of GS 4+3=7b tumours were stage pT3-4. In cases with PSA 10ng/ml or tumour extent >20% had higher scores (7b or higher). Cancers with tumour infiltration of 10ng/ml or tumour extent >20% had higher scores (7b or higher). Cancers with tumour infiltration of

DNA methylation regulates gene expression in normal cells. This epigenetic mechanism acts in at least two different ways: at global genomic level by targeting repetitive sequences distributed among the whole genome (LINEs, SINEs, satellite DNA, transposons) and at local level by targeting CpG islands in promoter regions. Both epigenetic mechanisms are involved in the carcinogenetic process; however, different evidences suggest that promoter hypermethylation occurring in genes involved in cellcycle regulation, DNA repair, cell signalling, transcription and apoptosis likely plays a prominent role. Opposite to genetic defects DNA hypermethylation is a reversible process that can be handled through “epigenetic drugs” in a wide spectrum of tumors. Along this line, recent data have demonstrated the ability of DNA hypomethylating agents to up-regulate and/or induce the expression of genes silenced by promoter hypermethylation in cancer. Particularly relevant seems the ability of these drugs to modulate the expression of genes coding for molecules crucial for tumor immunogenicity and immune recognition of neoplastic cells by host’s immune system, such as Cancer Testis Antigens, HLA class I molecules, costimulatory molecules. These evidences, coupled to the well-known cytotoxic, pro-apoptotic, and differentiating activities of epigenetic drugs, encourage to design and to develop new therapeutic strategies able to circumvent the immune escape of neoplastic cells and to potentiate the efficacy of immunotherapy in cancer patients. This review will provide an update on the most recent information about aberrant DNA methylation in cancer and on innovative therapeutic strategies of “epigenetic remodelling” of human malignancies, with particular attention to the immunologic and immunotherapeutic potential of this approach.

HuR, an ubiquitously expressed member of the Hu family, selectively binds and stabilizes AREcontaining mRNAs encoding proto-oncogenes, cell cycle regulators, cytokines and growth factors. The mechanism of HuR stabilization on target mRNAs is believed to be mediated through competition with destabilizing ARE-BPs. HuR is mainly localized within the cell nucleus and the nucleo-cytoplasmic shuttling of HuR is generally assumed as the initial and critical step of its stabilizing effects. A number of signaling pathways are believed to be involved in HuR shuttling. Due to the pivotal role played by HuR in stabilizing the mRNA of key factors or cytokines involved in carcinogenesis and subsequent progression, its implication and therapeutic potential in cancer have been investigated intensively since its discovery in 1996. This review discusses the role of HuR in the stabilization of key mRNAs and it’s the nucleo-cytoplasmic shuttling. The review also covers the current knowledge of HuR’s role in carcinogenesis, particularly its involvement in breast cancer, and the feasibility of using HuR as a therapeutic target for the treatment of breast cancer.