Ewing's sarcoma family of tumors (ESFTs) contains multiple tumors with similar histological and immunohistochemical features. ESFTs are small, round cell, highly malignant tumors that arise from the neuroectoderm of bone and extraskeletal soft tissue. Ewing's sarcoma is the second most common primary malignant bone cancer in children and adolescents, with the second decade of life being the most common age of diagnosis. In this article, we present a case of a young male who presented to the emergency department complaining of shortness of breath and cough and was later diagnosed with Ewing's sarcoma of the chest wall, which is also called Askin's tumor, and it is an extremely rare disease with only 17 cases reported in the literature.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

No description available.

O HHV-8 (herpesvírus 8 humano) é o agente etiológico do sarcoma de Kaposi (SK). Diferentemente dos outros herpesvírus, o HHV-8 é distribuído de modo não ubíquo ao redor do mundo. São sete os principais genótipos de HHV-8, de acordo com o padrão de variabilidade da ORF K1: A, B, C, D, E, F e Z. Estudos da variabilidade genética do HHV-8 poderão trazer melhores interpretações sobre o potencial patogênico dos genótipos de HHV-8 e das variações genotípicas funcionais. Dados sobre a variabilidade genética do HHV-8 no Brasil, em que o SK é associado ao HIV, permanecem escassos. Pelo nosso conhecimento, esse é o primeiro estudo que compara a variabilidade genética de HHV-8 em indivíduos infectados por HIV com SK e sem SK no Brasil. Objetivo. O estudo visou analisar a variabilidade genética do HHV-8 entre indivíduos infectados por HIV com SK e sem histórico de SK. Métodos. Sequências de DNA de HHV-8 foram investigadas em amostras criopreservadas de células mononucleares do sangue periférico a partir de 37 indivíduos infectados por HIV com SK (grupo 1); e de amostras de saliva de indivíduos sem SK (grupo 2), as quais foram selecionadas por meio da detecção positiva de DNA/ORF73/HHV-8 a partir de um total de 751 indivíduos. Dados demográficos e clínicos do estadio e evolução do SK, assim como parâmetros laboratoriais foram caracterizados. As análises moleculares e reconstruções filogenéticas foram baseadas nas ORFs K1 e K12 do HHV-8. Resultados. Foram obtidas sequências de DNA dos loci K1 e/ou K12 de 75 indivíduos, 34 indivíduos do grupo 1 e 41 do grupo 2. O sistema de primers empregado foi capaz de detectar os genótipos A, B, C, F e amplo perfil de subgenótipos de K1/HHV-8. Os dados não mostraram associação de genótipos de HHV-8 com a presença de SK ou estadio de SK. Todavia, o subgenótipo B1 predominou naqueles em que não houve registro de piora de SK (p=0,04). Os subgenótipos B1 e C3 foram igualmente predominantes em ambos os grupos. As frequências do genótipos A foram de 24% e 12,2% e dos genótipos B e C foram de 34,1 e 35,3%, nos grupos 1 e 2, respectivamente. O genótipo F foi descrito pela primeira no Brasil, um caso de cada grupo. Um amplo perfil de subgenótipos de C no grupo 2 sem SK foi encontrado (C1, C2, C3, C5 e C7). Subgenótipos K1 C5 e C7, exclusivos do grupo 2 (7%), foram confirmados como recombinantes. Não houve variabilidade genotípica de HHV-8 em amostras biológicas diferentes do mesmo indivíduo em oito casos estudados. Sítios polimórficos (6/59) em regiões codificadoras de miRNA do locus K12 foram observados, sendo 70% presentes exclusivamente em sequências de HHV-8 do grupo com SK e protótipos de SK. Conclusão. Embora não houvesse associação entre genótipos de HHV-8 e presença ou estadio de SK, o subgenótipo B1 foi significativamente relacionado ao melhor prognóstico de SK. Alguns recombinantes foram observados no locus K1 de HHV-8 em indivíduos do grupo 2 sem SK. A presença de SNPs em regiões codificadoras de miR12-12 e miR12-10 predominou em sequências de HHV-8 de indivíduos com SK, grupo 1, e protótipos de SK epidêmico, endêmico e clássico. A escoha de primers foi importante para garantir a amplificação de todos os genótipos e amplo perfil de subgenotipos de HHV-8. / HHV-8 (Human Herpes Virus 8) is the etiological agent of Kaposi\'s sarcoma (KS). Unlike other herpesviruses, the distribution of HHV-8 is not so ubiquitous around the world. There are seven major HHV-8 genotypes, according to the variability pattern of the ORF K1: A, B, C, D, E, F and Z. Studies on the genetic variability of HHV-8 may help to better understand the pathogenic potential of HHV-8 genotypes and their functional genotypic variations. Data on the genetic variability of HHV-8 in Brazil, where KS is associated with HIV infected people, remain scarce. To our knowledge, this is the first study comparing the genetic variability of HHV-8 among HIV-infected individuals with KS and without KS in Brazil. Objective. The study aimed to analyze the genetic variability of HHV-8 among HIV-infected individuals with and without KS. Casuistry and Methods. HHV-8 DNA sequences were obtained from samples of cryopreserved peripheral blood mononuclear cells from 37 individuals infected with HIV-KS (group 1); and saliva from individuals without KS (group 2) who were selected by means of detection of positive DNA/ORF73/HHV-8 from a total of 751 individuals. Demographic and clinical data (stage and progression of KS), as well as laboratory parameters were characterized. Molecular analysis and phylogenetic reconstructions were based on sequences of the ORFs K1 and/or K12 of HHV-8. Results. K1 and/or K12 DNA sequences of HHV-8 were obtained from 75 subjects, 34 from group 1 and 41 from group 2. The primer system used was able to detect the genotypes A, B, C, F and a wide profile of HHV- 8 K1 subgenotypes. Data showed no association of genotypes with the occurrence of KS or with visceral KS either. However, subgenotype B1 predominated in individuals who did not report any progression of KS (p=0.04). Subgenotypes B1 and C3 were equally prevalent in both groups. Genotype A frequencies were 24 % and 12.2% and genotypes B and C were 34.1 and 35.3 % in groups 1 and 2, respectively. We also described here for the first time the genotype F of HHV-8 in Brazil. A wide profile of subgenotypes C (C1, C2, C3, C5 and C7) in the group without KS was found. HHV-8 K1 DNA sequences of group 2 (7%) belonging to subgenotypes C5 and C7 were confirmed as recombinants. Our findings did not show virus variability in the same patient in samples collected at different times or from different biological material in the eight cases studied here. There was no statistical difference regarding the presence/absence of a given SNP from locus K12 between groups with and without KS. However, there were a total of 6/59 polymorphic sites in coding regions of miRNA, 70% of which present only in the HHV-8 DNA sequence of group with KS and KS prototypes. Conclusion. Although there was no association between HHV-8 genotypes and the presence of KS and KS clinical stage, subgenotype B1 was significantly related to the absence of progression of KS. Some recombinants in K1/HHV-8 locus were observed in the group without KS. The presence of SNPs in coding regions of miR12-12 and miR12- 10 predominated in sequences of HHV- 8 of SK cases (group 1) and of epidemic, endemic and classic KS prototypes. The choice of primers was essential to ensure amplification of all HHV- 8 genotypes and wide profile de subgenotypes.

O Sarcoma de Aplicação Felino (SAF) possui características específicas pouco estudadas. A agressividade, com invasão profusa de tecidos adjacentes, bem como a presença de células anaplásicas em sua composição fazem do SAF um obstáculo a ser transposto, para obtenção do melhor tratamento. Sua etiologia e etiopatogenia ainda permanecem obscuras. O objetivo deste estudo consistiu em determinar fatores prognósticos para o paciente portador de SAF, caracterizando-o quanto ao seu peso e escore de condição corporal (ECC), à origem e diferenciação da neoplasia por ele apresentada, bem como caracterizar o microambiente perineoplásico quanto à presença de células inflamatórias. Foram selecionados 46 casos de SAF, sendo 31 casos novos de pacientes e 22 casos reavaliados. Foi encontrada diferença significativa entre o ECC (p=0,001) e peso (p<0,001) de pacientes que apresentaram SAF e os do grupo controle. Não foram encontradas diferenças entre gênero e raça. A localização mais comumente acometida foi a tóraco-abdominal (78%). Quanto à origem, a maioria dos SAF apresentou-se como originada em células fibroblastos. Enquanto não houve marcação para componentes inflamatórios (CD3 e CD68), houve para receptor de quinase (c-kit), para Cox-2 e FelV. Não houve influencia dos parâmetros avaliados com as variáveis clínicas e com a sobrevida total. Houve influencia estatisticamente significante quando comparamos a invasão observada no exame de tomografia computadorizada e a escolha do tratamento instituído (p=0,019). / The feline injection-site sarcoma (FISS) has specific characteristics which are little studied. FISS treatment is a hurdle to getting because of their aggressiveness, with profuse invasion of adjacent tissue, as well as the presence of anaplastic cells in its composition. Its etiology and pathogenesis remain obscure. This research aims to determine prognostic factors for patients with FISS, characterizing it as their weight and body condition score (BCS) and by the origin and differentiation of the tumor presented, along with characterizing the microenvironment peri-neoplastic for the presence of inflammatory cells. 46 cases of cats that presented FISS were assisted, with 31 new cases and 22 cases of patients reassessed. Statistical difference was found between the BCS (p=0,001) and weight (p<0,001) of FISS group and control group. No differences were found between gender and race. The region most commonly affected was the toraco-abdominal (78%). The FISS majority presented as fibroblasts origin, while no marking for inflammatory components (CD3 and CD68). There was marking for kinases cascade (c-KIT), for COX-2, and for FelV. There was no influence of the parameters evaluated with clinical variables and overall survival. There was a statistically significant influence when comparing the invasion observed in computerized tomographic examination and choice of the treatment (p = 0.019).

A mucosa bucal e a pele podem apresentar distúrbios vasculares proliferativos com comportamento biológico variado. Para classificar tais lesões foram adotados critérios etiopatogênicos e a avaliação desses critérios em grupos específicos de lesões angioproliferativas pode contribuir para o diagnóstico e tratamento das mesmas. O objetivo desse estudo retrospectivo foi caracterizar as principais lesões angioproliferativas da mucosa bucal, comparando-as com os mesmos tipos de lesões cutâneas, em relação aos achados histopatológicos e à expressão endotelial do fator de crescimento fibroblástico básico (FGFb) e do receptor p75 para neurotrofinas (p75NTR). Foram utilizados espécimes arquivados em blocos parafinados, divididos conforme o diagnóstico microscópico em seis grupos: (I) malformações venosas; (II) malformações linfáticas; (III) granulomas piogênicos; (IV) hemangiomas capilares; (V) hemangiomas arteriovenosos; (VI) sarcomas de Kaposi. Cada grupo foi subdividido em dois grupos quanto a sua localização bucal e cutânea. Os padrões microscópicos das lesões angioproliferativas bucais e cutâneas de nossa amostra são similares e podem ser classificadas sob os mesmos parâmetros. No entanto, alguns grupos apresentaram diferenças quanto ao estádio de maturação das lesões, em relação à incidência e principalmente em relação à expressão dos mediadores estudados. A diferença na expressão de FGFb e p75NTR, observada entre os grupos bucais e cutâneos dos mesmos tipos de lesões, permitiunos concluir que a evolução da angiogênese na mucosa bucal e na pele não é necessariamente mediado pelos mesmos fatores. Tais diferenças abrem perspectivas para pesquisas futuras e abordagens terapêuticas baseadas em tecnologias moleculares. / The oral mucosa and the skin may present proliferative vascular diseases with varied biological behavior. To classify such lesions, pathogenic criteria were adopted in specific groups of vascular lesions. The objective of this retrospective study was to characterize the main oral vascular lesions, comparing them with the same types of cutaneous lesions. The histopathologic findings were compared, as well as the endothelial expression of basic fibroblastic growth factor (FGFb) and p75 neurotrophin receptor (p75NTR). Paraffin embedded specimens were purchased and divided, according to microscopic diagnostic, into six groups: (I) venous malformations; (II) lymphatic malformations; (III) pyogenic granulomas; (IV) capillary hemangiomas; (V) arteriovenous hemangiomas; (VI) Kaposi sarcomas. Each group was divided in two subgroups of oral and cutaneous location. The microscopic data demonstrated that oral and cutaneous vascular lesions, in our sample, are similar and they can be classified under the same parameters. However, some groups presented differences related to maturation stage, incidence, and expression FGFb and p75NTR. These differences allowed us to conclude that the evolution of angiogenic process inside oral mucosa and in the skin, not necessarily is mediated by the same biological factors. Such differences open perspectives for future researches and therapeutic approaches, based on molecular technologies.

Genome sequencing is emerging as a powerful tool to identify the molecular mechanism of cancer progression. However, the software tools to define genomic and post-genomic mutations are just in its infancy. We have used a novel software algorithm to analyse the cancer genome by DNAseq and expressed cancer genome arising from transcription by RNAseq to define dominant sources of potentially expressed tumour-specific mutations and oncogenic targets. We focus primarily on the rare human pleomorphic sarcoma as a disease of high unmet clinical need but use a range of cancer models to accelerate the development of the pipeline. First, we applied next generation sequencing of whole exomes of tumour tissues and two matched normal tissues (blood and “normal” tumour adjacent tissue) from a small set of patients to define parameters for use of the new software. The approaches identified significant mutations in tumour relative to germline DNA, but also in normal adjacent tissue, relative to normal germline, consistent with known field cancerization. Thus, in setting up the larger sequencing screen in the subsequent set of twenty cancer pleomorphic sarcoma cancer patients, whole exome sequencing was performed on tumour tissue and their matched normal adjacent tissues, rather than germline blood derived DNA, to define truly tumour-specific mutations. This approach provided sets of recurrent non-synonymous mutations in tumour tissue such as a transmembrane protease and suggests potential therapeutic targets for future focus that are highly tumour specific in pleomorphic sarcoma. A major problem with using DNA genomics only to define drugable landscapes in cancer is that the tumour genome is static and the mutations do not reflect the expressed cancer landscape at the time of surgery. Thus, in a smaller subset of patients we also applied shotgun RNAseq to determine the number of expressed mutated genes. We defined within the parameters chosen, from 8-17% of the mutated genome is expressed as defined at the RNA level. However, to our surprise, there were an order of magnitude more RNA mutations that were not DNA encoded suggestive of RNA editing events. Each patient showed elevated RNA edits that were independent of each other suggesting a highly-patient, cancer-specific perturbation in the specificity of the RNA editing machinery. We thus developed a cancer cell model to validate the RNA-editing software and we found we could recapitulate some of the RNA edits observed in clinical tumour tissue, in particular the signalling kinase in the MAP kinase-kinase-kinase-kinase super-family. It was interesting that RNA edits can often cluster in exon-intron boundaries suggesting a link to splicing and allows us to begin to produce “rules” for RNA editing. These data provide future direction to understand the role of RNA editing, as well as DNA encoded mutations, as mutagenic events and possible drugable targets in cancer signalling. Lastly, novel or orphan mutant proteins observed in human cancers, whether from DNA encoded mutant proteins or from RNA-edited driven mutant protein synthesis require new tools and technologies to discover new oncogenic signalling mechanisms. We developed an SBP-tagged affinity purification method in combination with label-free SWATH mass spectrometry to identify a novel binding protein for the gain-of-function mutant protein in a key metastatic gene, ELMO1. This identified an elevated interaction with another oncogenic protein encoded by AGR2 gene and validates this proteomics discovery platform to further advance function of new mutated proteins. In conclusion, we have applied and validated newly emerging software to begin to interrogate cancer tissue from patients of unmet clinical need in order to define new mechanisms of cancer progression and to define possibly new or better drug targets for new therapies. The data identified highly recurrent genome encoded mutations in human pleomorphic sarcoma and a potentially novel, targetable landscape represented by RNA editing driven mutant protein production. This will provide a foundation for future work on making better choices to advance our ability to improve patient management in human pleomorphic sarcoma.

A mucosa bucal e a pele podem apresentar distúrbios vasculares proliferativos com comportamento biológico variado. Para classificar tais lesões foram adotados critérios etiopatogênicos e a avaliação desses critérios em grupos específicos de lesões angioproliferativas pode contribuir para o diagnóstico e tratamento das mesmas. O objetivo desse estudo retrospectivo foi caracterizar as principais lesões angioproliferativas da mucosa bucal, comparando-as com os mesmos tipos de lesões cutâneas, em relação aos achados histopatológicos e à expressão endotelial do fator de crescimento fibroblástico básico (FGFb) e do receptor p75 para neurotrofinas (p75NTR). Foram utilizados espécimes arquivados em blocos parafinados, divididos conforme o diagnóstico microscópico em seis grupos: (I) malformações venosas; (II) malformações linfáticas; (III) granulomas piogênicos; (IV) hemangiomas capilares; (V) hemangiomas arteriovenosos; (VI) sarcomas de Kaposi. Cada grupo foi subdividido em dois grupos quanto a sua localização bucal e cutânea. Os padrões microscópicos das lesões angioproliferativas bucais e cutâneas de nossa amostra são similares e podem ser classificadas sob os mesmos parâmetros. No entanto, alguns grupos apresentaram diferenças quanto ao estádio de maturação das lesões, em relação à incidência e principalmente em relação à expressão dos mediadores estudados. A diferença na expressão de FGFb e p75NTR, observada entre os grupos bucais e cutâneos dos mesmos tipos de lesões, permitiunos concluir que a evolução da angiogênese na mucosa bucal e na pele não é necessariamente mediado pelos mesmos fatores. Tais diferenças abrem perspectivas para pesquisas futuras e abordagens terapêuticas baseadas em tecnologias moleculares. / The oral mucosa and the skin may present proliferative vascular diseases with varied biological behavior. To classify such lesions, pathogenic criteria were adopted in specific groups of vascular lesions. The objective of this retrospective study was to characterize the main oral vascular lesions, comparing them with the same types of cutaneous lesions. The histopathologic findings were compared, as well as the endothelial expression of basic fibroblastic growth factor (FGFb) and p75 neurotrophin receptor (p75NTR). Paraffin embedded specimens were purchased and divided, according to microscopic diagnostic, into six groups: (I) venous malformations; (II) lymphatic malformations; (III) pyogenic granulomas; (IV) capillary hemangiomas; (V) arteriovenous hemangiomas; (VI) Kaposi sarcomas. Each group was divided in two subgroups of oral and cutaneous location. The microscopic data demonstrated that oral and cutaneous vascular lesions, in our sample, are similar and they can be classified under the same parameters. However, some groups presented differences related to maturation stage, incidence, and expression FGFb and p75NTR. These differences allowed us to conclude that the evolution of angiogenic process inside oral mucosa and in the skin, not necessarily is mediated by the same biological factors. Such differences open perspectives for future researches and therapeutic approaches, based on molecular technologies.

According to the UNAIDS global report 2013, an estimated 6.1 million people are living with human immunodeficiency virus (HIV) in South Africa. The incidence of Kaposi sarcoma (KS) has increased dramatically since the Acquired Immunodeficiency Syndrome (AIDS) epidemic. Of the estimated 66 200 cases of KS worldwide, 58 800 are thought to have occurred in SSA (Parkin 2002). However, there remains a paucity of published data about KS from South Africa. This retrospective study was conducted to describe the epidemiology of KS at Chris Hani Baragwanath Academic Hospital (CHBAH) and to determine possible links among the CD4 counts, intensity and distribution of human herpes virus 8 latency- associated nuclear antigen 1 (HHV8 LNA-1) immunohistochemical staining and the stage of KS. Nine hundred and thirty eight histopathology reports of KS diagnosed in 901 patients at CHBAH between 2005 and 2009 were reviewed and demographic data (age, gender, topographic site, CD4 count, HIV status, KS stage, HHV8 LNA-1 staining, concomitant pathology) were recorded. The H&E stained sections and HHV8 LNA-1 immunostains of a cohort of 127 cases were subsequently reviewed and categorised with regard to intensity and distribution of staining. The male:female ratio was 1,2:1. The mean age was 36,8 years (standard deviation {SD} 10,2 years) and the median CD4 count 127,5 cells/mm3 (quartile range {QR} 184,5 cells/mm3). Lower limb skin biopsies accounted for 49,6% of cases. Concomitant pathology was seen in 4,6% of cases. Infections and inflammatory dermatoses were the most frequently diagnosed concomitant pathology in cutaneous biopsies. Paediatric, visceral and endemic KS accounted for only limited proportions of cases (1,44% of patients; 1,4% and 1,3% respectively). There was a significant difference in the distribution of HHV8 LNA-1 staining in patch versus nodular KS (p = 0,011). The CD4 counts were not predictive of KS Page | v stage (p = 0,701) or the intensity (p = 0,877) and distribution (p = 0,846) of HHV8 LNA-1 immunohistochemical staining. This study highlights the epidemiology of KS and the variability in HHV8 LNA-1 immunohistochemical staining across CD4 counts and stages of KS.

Thesis (Ph. D.)--Cornell University, January, 2007. / Vita. Includes bibliographical references (leaves 146-195).