Expression of histone methyltransferases as novel biomarkers for renal cell tumor diagnosis and prognostication

Renal cell tumors (RCTs) are the most lethal of the common urological cancers. The widespread use of imaging entailed an increased detection of small renal masses, emphasizing the need for accurate distinction between benign and malignant RCTs, which is critical for adequate therapeutic management. Histone methylation has been implicated in renal tumorigenesis, but its potential clinical value as RCT biomarker remains mostly unexplored. Hence, the main goal of this study was to identify differentially expressed histone methyltransferases (HMTs) and histone demethylases (HDMs) that might prove useful for RCT diagnosis and prognostication, emphasizing the discrimination between oncocytoma (a benign tumor) and renal cell carcinoma (RCC), especially the chromophobe subtype (chRCC). We found that the expression levels of three genes-SMYD2, SETD3, and NO66-was significantly altered in a set of RCTs, which was further validated in a large independent cohort. Higher expression levels were found in RCTs compared to normal renal tissues (RNTs) and in chRCCs comparatively to oncocytomas. SMYD2 and SETD3 mRNA levels correlated with protein expression assessed by immunohistochemistry. SMYD2 transcript levels discriminated RCTs from RNT, with 82.1% sensitivity and 100% specificity (AUC=0.959), and distinguished chRCCs from oncocytomas, with 71.0% sensitivity and 73.3% specificity (AUC: 0.784). Low expression levels of SMYD2, SETD3, and NO66 were significantly associated with shorter disease-specific and disease-free survival, especially in patients with non-organ confined tumors. We conclude that expression of selected HMTs and HDMs might constitute novel biomarkers to assist in RCT diagnosis and assessment of tumor aggressiveness.

Antidepressant induced excessive yawning and indifference

Introduction Antidepressant induced excessive yawning has been described as a possible side effect of pharmacotherapy. A syndrome of indifference has also been described as another possible side effect. The frequency of those phenomena and their physiopathology are unknown. They are both considered benign and reversible after antidepressant discontinuation but severe cases with complications as temporomandibular lesions, have been described. Methods We report two unprecedented cases in which excessive yawning and indifference occurred simultaneously as side effects of antidepressant therapy, discussing possible physiopathological mechanisms for this co-occurrence. Case 1: A male patient presented excessive yawning (approximately 80/day) and apathy after venlafaxine XR treatment. Symptoms reduced after a switch to escitalopram, with a reduction to 50 yawns/day. Case 2: A female patient presented excessive yawning (approximately 25/day) and inability to react to environmental stressors with desvenlafaxine. Conclusion Induction of indifference and excessive yawning may be modulated by serotonergic and noradrenergic mechanisms. One proposal to unify these side effects would be enhancement of serotonin in midbrain, especially paraventricular and raphe nucleus.

Extensive subcutaneous emphysema as a presentation of ischemic colitis

Introduction . Subcutaneous emphysema is usually benign and self-limited; however, it may be associated with a life-threating situation. Case Report . An elderly woman with progressive malaise with extensive subcutaneous emphysema (cervical to abdominal wall) was observed at the emergency department. Colonic perforation was diagnosed and the patient underwent surgery. Intraoperatively, necrosis and perforation of the sigmoid colon into the retroperitoneum were found and a Hartmann procedure was performed. Conclusion . Cervical and thoracic subcutaneous emphysema may be the first sign of intra-abdominal lesion.

Contributo para a compreensão das causas de exclusão de ilicitude e da culpa no crime de acesso ilegítimo

Dissertação de mestrado em Direito e Informática

An efficient Azorean thermophilic consortium for lignocellulosic biomass degradation

[Excerpt] Lignocellulosic plant biomass is being envisioned by biorefinery industry as an alternative to current petroleum platform because of the large scale availability, low cost and environmentally benign production. The industrial bioprocessing designed to transform lignocellulosic biomass into biofuels are harsh and the enzymatic reactions may be severely compromised reducing the production of fermentable sugars from lignocellulosic biomass. Thermophilic bacteria consortium are a potential source of cellulases and hemicellulases adapted to extreme environmental conditions, which can be exploited as a new source for the development of more robust enzymatic cocktails. (...)

Evaluation of the heart rate and arrhythmias following the Maze procedure for chronic atrial fibrillation

PURPOSE: To assess the presence and the prevalence of arrhythmias and the variability of the heart rate in the medium-term postoperative period following the maze procedure for chronic atrial fibrillation (AF). METHODS: Seventeen patients with a mean age of 51.7±12.9 years, who previously underwent the maze procedure without cryoablation for chronic atrial fibrillation, were evaluated with the 24 hour electrocardiogram (ECG) - Holter monitoring from the 6th month after the operation. Valvular and coronary procedures were concomitantly performed. RESULTS: The mean heart rate during Holter monitoring was 82±8bpm; the maximal heart rate was 126 ± 23bpm and the minimal heart rate 57±7bpm. Sinus rhythm was found in 10 (59%) patients and atrial rhythm was found in 7 (41%). Supraventricular extrasystoles had a rate of 2.3±5.5% of the total number of heartbeats and occurred in 16 (94%) patients. Six (35%) patients showed nonsustained atrial tachycardia. Ventricular extrasystoles, with a rate of 0.8±0.5% of the total heartbeats, occurred in 14 (82%) patients. The chronotropic competence was normal in 9 (53%) patients and attenuated in 8 (47%). The atrioventricular conduction (AV) was unchanged in 13 (76%) patients and there were 4 (24%) cases of first degree atrioventricular block (AVB). CONCLUSION: After the maze procedure, the values for the mean heart rate, AV conduction and chronotropic competence approach the normal range, although some cases show attenuation of the chronotropic response, first degree AV block or benign arrhythmias.

Cardiac tamponade in systemic lupus erythematosus. Report of four cases

OBJECTIVE: To report and assess the incidence of cardiac tamponade in systemic lupus erythematosus as a cardiac manifestation of the disease. METHODS: We reviewed the medical records of 325 patients diagnosed with systemic lupus erythematosus according to the American Rheumatism Association and their complementary laboratory tests compatible with cardiac tamponade. RESULTS: In the 325 medical recors reviewed, we found 108 patients with pericardial effusions corresponding to 33.2% of the total and 54% of the patients studied in the active phase of the disease. Clinical assessment and transthoracic echocardiogram allowed the clinical diagnosis of cardiac tamponade in only 4 (1.23%) patients, 3 of whom were females, white, with ages ranging from 25 to 44 years. The pericardial fluid was hemorrhagic or serosanguineous with high levels of FAN and positivity for LE cells. In the treatment, we successfully used pericardicentesis associated with high doses of corticosteroids. In clinical and laboratory follow-up performed for a period of 3 years, neither recrudescence of the pericardial effusion nor evolution to constriction occurred. CONCLUSION: Even though rare (1.23%), cardiac tamponade in patients with systemic lupus erythematosus has a benign evolution when properly treated, according to our experience.

The role of macroH2A1 in prostate carcinogenesis

Dissertação de mestrado em Genética Molecular

Chagas' heart disease: evolutive evaluation of electrocardiographic and echocardiographic parameters in patients with the indeterminate form

OBJECTIVE: To identify and associate potential electrocardiographic and echocardiographic changes in patients with the indeterminate form of Chagas' disease during long-term follow-up. METHODS: One hundred sixty patients underwent standard electrocardiography and two-dimensional guided M-mode echocardiography for left ventricular ejection fraction determination. Patients were followed up for 98.6±30.4 months, undergoing repeat electrocardiographic studies at 6-month intervals and echocardiographic studies at 12-month intervals. RESULTS: Based on the electrocardiographic findings, the patients were divided into group I, 125 patients (78.6%) with normal electrocardiograms throughout follow-up, and group II, 34 patients (21.3%) who developed electrocardiographic changes. Group II was further divided into group IIA (9 patients, 5.6%) with permanent electrocardiographic changes, group IIB (14 patients, 8.8%) with transitory electrocardiographic changes, and group IIC (11 patients, 6.9%) with changes appearing only on the final electrocardiogram. Left ventricular ejection fractions remained normal in the entire population studied and did not differ among groups. CONCLUSION: The indeterminate form of Chagas' disease clearly represents a benign condition with a favorable long-term prognosis. Although some patients develop electrocardiographic changes, left ventricular systolic function is well preserved.

Estudios de quimioprevención en: 1) Modelo NMU de carcinogénesis de páncreas de rata. 2) Osteosarcoma inducido por NMU

El cáncer es una enfermedad común de causa de muerte en el mundo entero. Si bien el éxito en el tratamiento de casi todas las formas de esta enfermedad depende de su diagnóstico temprano seguido de resección quirúrgica, los enfoques en investigación sobre compuestos inhibitorios de la carcinogénesis podrían ser aplicados a implementar estrategias de quimioprevención poblacionales. Se sabe que el carcinoma de páncreas exocrino ocupa el cuarto lugar como causa de muerte en el hombre en los Estados Unidos. Se presupone que su incidencia es similar en nuestro país. La causa del cáncer de páncreas es desconocido en la mayoría de los casos. Los estudios epidemiológicos si bien no dan datos certeros tienden a señalar que componentes de la dieta son factores de riesgo en la génesis del cáncer de páncreas (Rogers y Longnecker, 1988). Se conoce que el cáncer de páncreas es clínicamente silencioso y sólo manifiesta síntomas tardíamente, cuando ya se ha producido la invasión y metástasis (Pour, 1991). De ahí que haya interés por desarrollar modelos multifactoriales que puedan aportar datos para comprender las causas y el mecanismo de desarrollo del cáncer en el hombre. Trabajos realizados en el laboratorio del Programa Centro de Biología Celular y Desarrollo (CEBYD-CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Director Dr. Benito Monis, han permitido establecer que la nitrosometilurea (NMU) induce la proliferación de poblaciones celulares exocrinas del páncreas de rata con la formación de estructuras nodulares atípicas, múltiples, de crecimiento expansivo que aumenta en número y tamaño con la edad, a los que identificaron con la sigla FACH (Focal Acinar Cell Hyperplasia). Este proceso hiperplásico es primariamente del epitelio acinar, aunque se han observado con notable menor frecuencia la aparición de estructuras ductulares neoformadas y aislados focos microscópicos de células con apariencia de hepatocitos (transdiferenciación o metaplasia hepatocítica). Los nódulos son con mayor frecuencia acidófilos aunque pueden observarse focos basófilos. Estos últimos son de dimensión microscópica. Excepcionalmente se han encontrado lesiones cancerosas en páncreas en el numeroso grupo de ratas inyectadas con NMU y estudiadas luego de la muerte natural en el laboratorio, por lo cual se infiere que las lesiones proliferativas que el NMU induce en páncreas de rata no posee tendencia a la transformación maligna. Este es el concepto central en las investigaciones sobre carcinogénesis pancreática en curso en el laboratorio CEBYD y sobre el cual proponemos el presente plan de trabajo. Además, hemos aislado células de un osteosarcoma inducido en la rata por la NMU (en curso) que han sido mantenidas tanto "in vivo" como "in vitro" para su caracterización y para el estudio de agentes inhibitorios de la carcinogénesis. El propósito del presente plan de trabajo es probar posibles agentes quimioterapéuticos en ambos modelos experimentales desarrollados en nuestro laboratorio.

Rol de la infección bacteriana en el desarrollo y progresión del cáncer prostático. Influence of bacterial infection in the development and progression of prostatitic cancer.

En la patogénesis del cáncer, factores microambientales como la inflamación están estrechamente vinculados al desarrollo y crecimiento tumoral, sustentando la clásica hipótesis de Virchow del origen del cáncer en sitios de inflamación crónica, la cual incitaría a carcinogénesis por múltiples factores. Estudios previos en este laboratorio evidenciaron en un modelo de prostatitis bacteriana agudo con E. coli, profundos cambios estromales semejantes al "estroma reactivo", con predominio de miofibroblasto, que se genera en el cáncer. En correlación, existe abundante evidencia obtenida en modelos experimentales animales confirmando que el microambiente estromal en el cual se desarrollan los tumores epiteliales influencia profundamente la progresión tumoral. El rol protagónico del estroma del huésped en el crecimiento neoplásico, también se ha demostrado inoculando la misma línea tumoral en diferentes tejidos y analizando su comportamiento en comparación con su implantación en el sitio anatómico original del tumor (implante ortotópico); otro factor clave en la repuesta del huésped al tumor está dado por el infiltrado de células inmunes que puede favorecer o limitar el crecimiento tumoral de acuerdo al perfil de citoquinas que secreten. Teniendo en cuenta estos antecedentes, este proyecto tiene como Objetivo General estudiar la influencia de la infección bacteriana crónica en la inducción y evolución del cáncer prostático. Para ello trabajaremos in vivo con dos formas de formas de Tumores Prostáticos, un Tumor Inducido por combinación del carcinógeno N-methyl-N-nitrosourea (MNU) y testosterona; el segundo mediante Transplante Ortotópico de células tumorales prostáticas MAT-LU. En ambos modelos se inducirá previamente una prostatitis bacteriana, a fin de estudiar los efectos de la prostatitis en la inducción del tumor en el primer modelo, y en la implantación de las células tumorales en el segundo. También se inducirá prostatitis después de establecido el tumor por ambos procedimientos, a fin de determinar si la prostatitis bacteriana modula la progresión neoplásica. Finalmente, proponemos un modelo in vitro que permita estudiar la interacción tumor/estroma separado de la influencia del sistema inmune. A tal fin se utilizarán co-cultivos combinando células tumorales con estromales modificadas de diferente modo. La Inducción de Tumores Prostáticos se realizará en ratas de la cepa Wistar adultas, en las cuales se inducirán lesiones displásicas y neoplásicas siguiendo protocolos de carcinogénesis prostática por MNU, para lo cual es necesario el tratamiento previo con acetato de ciprosterona y propionato de testosterona, seguido por administración crónica de testosterona. Los estudios con Transplante Ortotópico de células tumorales se realizarán en ratas Copenhagen. La influencia de la infección bacteriana en el desarrollo tumoral será investigada inyectando E. coli intraprostáticamente: Se realizará Análisis Macroscópico, de Parámetros Morfológicos de las lesiones tumorales, grado de malignidad, extensión e invasión de las lesiones de acuerdo a consensos internacionales, y Bioquímicos mediante análisis de la expresión, por IHQ y WB, de fosfatasa ácida, citoqueratina 8, Prostatic Binding Protein, PTEN (gen supresor tumoral) y el receptor de Andrógenos, todos parámetros de actividad y de transformación celular. También se evaluará apoptosis por TUNEL y proliferación celular. Los cambios del compartimiento estromal en respuesta al implante tumoral y la influencia de la inflamación bacteriana se evaluarán mediante análisis morfológico e inmunocitoquímico, caracterizando el fenotipo de las poblaciones celulares con a-actina, vimentina, calponina y tenascina. Se espera que los resultados aporten evidencias acerca de las interacciones bidireccionales entre células neoplásicas prostáticas y su entorno estromal, que en un futuro puedan servir como base para establecr estrategias para prevenir y/o modificar el crecimiento neoplásico.

Caracterización funcional de zeb1 en células en diferenciación y metastásicas. Functional characterization of zeb1 in differentiating and metastatic cells

IDENTIFICACIÓN ZEB1 (Zinc Finger E-box Binding Homeobox) es un factor de transcripción funcionalmente asociado con la diferenciación de células como miocitos, neuronas, células de sostén y linfocitos T, además de estar involucrado en la Transición Epitelial-Mesenquimatosa (EMT) de los tumores sólidos epiteliales. Aún no se ha revelado en profundidad la participación de ZEB1 en los procesos de proliferación y diferenciación en los que participa. Estamos interesados en los mecanismos de regulación de ZEB1 y los factores que intervienen en los procesos de diferenciación y transformación celular. HIPÓTESIS 1. Las vías de señalamiento regulan el estado de fosforilación y la función de ZEB1 en la célula normal, el cual se desregularía en la célula neoplásica llevando a cambios en la función normal de ZEB1 y consecuentemente a metástasis. 2. IGF-1 es la señal que, en asociación con el supresor de tumores CCN6, juega un rol causal en la regulación de ZEB1 y esto a su vez en la metástasis del cáncer de mama. OBJETIVO GENERAL: establecer el rol funcional de ZEB1, su interrelación con otros factores y su regulación en los procesos de diferenciación y transformación celular. OBJETIVOS ESPECIFICOS (incluye Materiales y Métodos) 1. Estudiar la participación de vías de señalización sobre la función biológica de ZEB1 en células normales y neoplásicas. Analizaremos la participación de señales intracelulares en la fosforilación de ZEB1 por experimentos de ganancia/pérdida de función de la vía (por uso de inhibidores farmacologicos, mutantes silenciadoras y siRNAs), lo cual sera evaluado en EMSAs, ChIP, transfecciones, inmunofluoresc, etc. 2. Estudiar el rol de IGF-1 y CCN6 sobre la expresión y el estado de fosforilación de ZEB1 en tumores mamarios benignos, no invasivos e invasivos y metastatizantes. A) Se estudiará la expresión y localización subcelular de ZEB1 en líneas celulares de cáncer mamario y en xenotransplantes de ratón con variada expresión de CCN6. B) Investigar la relevancia de la fosforilación de ZEB1 mediada por IGF-1 en el EMT por experimentos con ganancia/pérdida de función. RESULTADOS ESPERADOS Esperamos poder delinear la/s vía/s de señalización intracelular que fosforilan ZEB1 y así conocer sobre la regulación del mismo. Podremos establecer algunas bases para entender la biología básica del cáncer de mama e identificar blancos terapéuticos. IMPORTANCIA Un amplio conocimiento de los factores de transcripción y sus vías de señalamiento es necesario para el desarrollo tanto de pruebas diagnósticas como para la identificación de nuevos blancos terapéuticos para neoplasias. De modo que resulta de gran importancia clínica determinar el rol de ZEB1, sus proteínas y vías reguladoras en el proceso de oncogénesis. El desarrollo del proyecto prevé la formación de dos tesistas. Se continuaran colaboraciones con dos grupos extranjeros y se iniciara una tercera. ZEB1 (Zinc Finger E-box Binding Homeobox) is a transcription factor involved in cell differentiation and Epithelial Mesenchymal Transition (EMT) of epithelial tumors. We are interested in the study of mechanisms of regulation (pre and post transcriptional). S.A.1. To investigate post translational mechanisms of ZEB1 regulation in normal and cancer cells. We will analyze the involvement of intracellular signals in phosphorylation of ZEB1 by gain- and lost-of-function experiments. S.A.2. A) To determine the role of IGF-1 signaling and CCN6 in regulating the expression of hypo- and hyperphosphorylated forms of ZEB1 in benign and malignant breast cell lines and in xenograft mouse models by overexpressing and inhibiting CCN6 in breast cancer cells. B) To investigate the relevance of CCN6-mediated ZEB1 phosphorylation to EMT, breast cancer invasion and metastasis. The role of CCN6 on ZEB1 phosphorylation and regulation of E-cadherin, induction of EMT, invasion and metastasis of breast cells will be investigated using gain- and loss-of-function experiments.

Mortality and Embolic Potential of Cardiac Tumors

Background: Cardiac tumors are rare, mostly benign with high embolic potential. Objectives: To correlate the histological type of cardiac masses with their embolic potential, implantation site and long term follow up in patients undergoing surgery. Methods: Between January 1986 and December 2011, we retrospectively analyzed 185 consecutive patients who underwent excision of intracardiac mass (119 females, mean age 48±20 years). In 145 patients, the left atrium was the origin site. 72% were asymptomatic and prior embolization was often observed (19.8%). The diagnosis was established by echocardiography, magnetic resonance and histological examination. Results: Most tumors were located in the left side of the heart. Myxoma was the most common (72.6%), followed by fibromas (6.9%), thrombi (6.4%) and sarcomas (6.4%). Ranging from 0.6cm to 15cm (mean 4.6 ± 2.5cm) 37 (19.8%) patients had prior embolization, stroke 10.2%, coronary 4.8%, peripheral 4.3% 5.4% of hospital death, with a predominance of malignant tumors (40% p < 0.0001). The histological type was a predictor of mortality (rhabdomyomas and sarcomas p = 0.002) and embolic event (sarcoma, lipoma and fibroelastoma p = 0.006), but not recurrence. Tumor size, atrial fibrillation, cavity and valve impairment were not associated with the embolic event. During follow-up (mean 80±63 months), there were 2 deaths (1.1%) and two recurrences 1 and 11 years after the operation, to the same cavity. Conclusion: Most tumors were located in the left side of the heart. The histological type was predictor of death and preoperative embolic event, while the implantation site carries no relation with mortality or to embolic event.

Reticulo-endotheliomatose maligna

A spindle-cell sarcoma (fig. 5) apparently originating from the dura (fig. 4) was found at the autopsy of a male, mulato, 17 years of age. The bones of the skull (occipital and both parietals) were penetrated and destroyed (fig. 1 and 2). The nervous tissue was not penetrated, the only change in the brain being a depressed area where the tumor was included. Metastatic nodules were found in the liver (fig. 3),hepatic lymphnodes (fig. 14), spleen (fig. 12) and suprarenal bodies (fig. 15). The structure, however, in all those different locations was that of a typical endothelioma (figs. 8, 11 and 13). The cells are of large and moderate size, of polyhedral form, with vesicular nuclei, diminutive nucleoli and clear cytoplasm. (Figs. 6 and 8). They are arranged about a central lumen which represents a rudimentary vessel (figs. 9 and 13). Other areas are composed of cells without concentric arrangement (figs. 4 and 10). In small areas, the colums of liver cells are marginated in one side by typical sinusoids, while in the other side tumor cells arranged about a narrow lumen are seen suggesting a pathological (neoplastic) sinusoid (figs. 7 and 9). The case is considered as a multiple diffuse endothelioma. The origin of the tumor is referred to the reticulo-endothelial apparatus of the liver, the spleen, the suprarenal bodies and the lymph nodes, the structure being rather uniform in those organs. In the dura, the endothelioma reproduces the structure and presents the general character of a fibroblastic sarcoma; in some places, however, the structure of endothelioma could be found (fig.6). It corresponds to the reticulo-endotheliomatosis maligna according to Puhr's grouping of progressive changes in the reticulo-endothelial apparatus which is a follows: 1. HYPERPLASTIC - 1. Mnnocytic leukemia. 2. a) Aleukemic reticulosis (Goldschmid and Isaac). b) Idiopathic sarcoma of skin (Kaposi). c) Cutaneous sarcoid (Spiegler). 3. Secretory reticulosis. a) Gaucher's disease. b) Generalized xanthomatosis. c) Spleno-hepatomegaly with lipoidic cells (Pick). II. BLASTOMATOSUS OR NEOPLASTIC - 1. Benign - a) Circumscribed tumors. a) Epulis sarcomatosa; b) Benign giant-cells sarcoma of the bone - marrow of long bones. b) Generalized brown tumors of osteitis fibrosa. 2. Malignant - a) Circumscribed haemangio - endothelioma (reticulo- endothelioma (maligum). of {liver, spleen, bone-marrow. b) Generalized haemangio-endotheliomatosis (reticulo-endotheliomatosis maligna) (Grabowski).

Sôbre a hiperplasia celular no mixoma infectuoso do coelho

Definite hyperplasia of cells occurs in the skin lesions of the infectious myxoma of rabbits, more visible in such stages in which the intercellular basophilic substance is rather scanty (fig. 2). The increase in number of cells is the result of simplified forms of mitosis (modified type of mitosis, pseudoamitosis) which might readily be mistaken for amitosis in their final stages. Budding (figs. 20, 28, 29, 30) as well as constriction of the nucleus (figs. 18, 31, 32), and the formation of giant-cells (figs. 33, 34) are not rare. During the entire process the nuclear membrane does not desintegrate as in typical mitosis. Division of the cytoplasm following division of the nucleus has been demonstrated (fig. 17). Typical mitosis is practically absent. The cells which undergo hyperplasia present remarkable changes in their dimension, shape, and structure. The nucleus and cell-body are considerably enlarged (figs. 6, 7, 8). The shape of the nucleus is modified (figs. 8, 10, 15). Hypertrophy of nuclein, either as an intranuclear network (spireme?, figs. 9, 23), or in the form conspicuous, deeply staining masses which appear not to be homogeneous but to be composed of small particles closely clumped ("mulberries"?, figs. 12, 13, 14, 25, 26) occurs in most cells. While some of these pictures are probably related to necrosis of the cells as started by most of the previous workers, it is lekely that some of them may represent developmental stages of the modified mitosis (pseudoamitosis) here reported. In fact, fine cytological details not ordinarily preserved in necrotic cells (figs. 35, 36, 37) may be demonstrated in the socalled myxoma-cells subtted to approved cytological methods of study (fixation in B-15 and P. F. A.-3, staining in iron-hematoxylin).