INTRODUÇÃO: O mixofibrossarcoma (MFS) é um dos mais frequentes sarcomas de partes moles (SPM) em idosos que afeta principalmente as extremidades. Historicamente, é um grupo de tumores heterogêneos. Clinicamente está caracterizado por apresentar uma alta incidência de recorrência local (RL) e um conhecimento limitado sobre a sua capacidade de metástase. O índice de RL após a ressecção cirúrgica é relativamente maior quando comparado com outros SPM. Não existe um consenso em como identificar os pacientes com maior risco. O objetivo deste estudo foi analisar os fatores de prognóstico dos pacientes diagnosticados com MFS em uma instituição única. MÉTODOS: Foram analisados retrospectivamente os prontuários de 75 pacientes com diagnóstico confirmado de MFS nas extremidades, que foram submetidos a tratamento cirúrgico, nos últimos 25 anos. Comparamos idade, sexo, tamanho e localização do tumor, grau histológico segundo a Fédération Nationale des Centres de Lutte Contre le Cancer (FNCLCC) e o estádio segundo a American Joint Committee on Cancer (AJCC). A média de idade foi 49,7 anos. A localização foi: membro superior (25,4%), membro inferior (66,6%) e pelve (8%). Os pacientes apresentaram baixo, intermediário e alto grau, em: 29,3%, 24% e 46,7% dos tumores, respectivamente. Pelo tamanho foram categorizados em: =/< 5 cm (13,3%), > 5 e <10 cm (42,7%), > 10 e <15 cm (9%) e =/> 15 cm (18,7%). Em total, 26,7% receberam radioterapia pós-operatória. As margens foram livres em 76% e comprometidas em 24%. A análise da regressão de Cox bivariada foi utilizada para determinar as associações entre os fatores clínicos e de tratamento com a RL. RESULTADOS: O tempo médio de seguimento foi 30.7 meses. O 26,7% dos pacientes apresentaram RL. Metástase foi reportada em 27 (36%) pacientes. O local mais comum de metástase foi: pulmão (92,6%) e gânglios linfáticos (18,5%). O tempo médio de sobrevida dos pacientes com metástase foi 21,2 meses. Os fatores preditivos para RL foram: margens comprometidas (hazard ratio 5.47, 95% intervalo de confiança, 2.23-13.40, P < 0.001) e metástase (hazard ratio 10.24, 95% intervalo de confiança, 3.53-26.68, P < 0.001). Os fatores preditivos da sobrevida livre de RL foram: grau histológico, margens comprometidas (hazard ratio 3.18, 95% intervalo de confiança, 1.51-6.70, P =0.001), e metástase (hazard ratio 15.23, 95% intervalo de confiança, 5.57-41.61, P= 0.001). Os fatores preditivos de sobrevida em geral foram: RL (hazard ratio 5.13, 95% intervalo de confiança, 2.15-12.24, P < 0.001), e metástase (hazard ratio 540.97, 95% intervalo de confiança, 5.04-58112.03, P < 0.001). CONCLUSÃO: As margens cirúrgicas comprometidas e a metástase estão diretamente associadas com a RL. O grau histológico do tumor, as margens comprometidas, a RL e a metástase, são fatores de pior prognóstico no MFS / BACKGROUND AND AIMS: Myxofibrosarcoma (MFS) is one of the most common soft tissue sarcomas (STS) in elderly patients and it primarily affects the extremities. They are a historically heterogeneous group of tumors. The clinical course of MFS is characterized by a high incidence of local recurrences (LR), but knowledge about distant metastasis is sparse. MFS is reported to have a higher risk of LR following definitive surgical excision relative to other STS. There is no agreement on how to identify patients at major risk. The objectives of this study were to analyze the prognostic factors and outcomes of patients with MFS treated at a single institution. METHODS: We retrospectively reviewed the records of 75 patients with pathologically confirmed MFS of the extremities who underwent surgery in the last 25 years. We compared the age, sex, tumor size and location, Fédération Nationale des Centres de Lutte Contre le Cancer (FNCLCC) grade and the American Joint Committee on Cancer (AJCC) stage. Median age was 49.7 years (range, 1 to 88 y). Site of disease was: upper extremity (25.4%), lower extremity (66.6%) and pelvic (8%). Patients had low, intermediate and high-grade, in: 29.3%, 24% and 46.7% of tumors, respectively. Tumors were categorized as =/< 5 cm (13.3%), > 5 and < 10 cm (42.7%), > 10 and < 15 cm (9%) and =/> 15 cm (18.7%). In total, 26.7% received postoperative radiotherapy. All patients underwent surgery. Margins were negative in 76% and positive in 24%. Bivariate Cox regression analysis was utilized to determine associations between clinical and treatment factors with LR. RESULTS: Median follow-up time was 30.7 months (range, 1.8 to 383.8 m). We found a 26.7% of LR. Distant metastasis was reported in 27 (36%) patients. The most common sites of metastasis were: lung (92.6%) and lymph nodes (18.5%). The overall survival rate in patients with metastasis was 21.2 months (range, 4.8 to 114.8 m). Predictors of LR were: positive margins (hazard ratio 5.47, 95% confidence interval, 2.23-13.40, P < 0.001) and distant metastasis (hazard ratio 10.24, 95% confidence interval, 3.53-26.68, P < 0.001). Predictors of overall survival free of LR were: grade, positive margins (hazard ratio 3.18, 95% confidence interval, 1.51-6.70, P =0.001), and distant metastasis (hazard ratio 15.23, 95% confidence interval, 5.57-41.61, P= 0.001). Predictors of overall survival were grade, LR (hazard ratio 5.13, 95% confidence interval, 2.15-12.24, P < 0.001), and distant metastasis (hazard ratio 540.97, 95% confidence interval, 5.04-58112.03, P < 0.001). CONCLUSION: In this institutional series of MFS, positive margins and distant metastasis were significantly associated with a higher risk of LR. Tumor grade, LR, positive margins and distant metastases were significant predictors of overall survival poor prognosis

Introdução: O carcinoma espinocelular (CEC) representa 7% de todos os novos casos de câncer no mundo, sendo o carcinoma espinocelular o tipo mais frequente. Tanto o comportamento biológico quanto o crescimento dos tumores devem ser melhores entendidos, uma vez que a sobrevida dos pacientes apresentou discreta melhora nas últimas décadas. Os modelos PDX foram desenvolvidos para estudar a biologia tumoral e principalmente os mecanismos de crescimento e proliferação através da manutenção da arquitetura e microambiente tumoral do tumor original. Os retinóides possuem a capacidade de restaurar o crescimento e a diferenciação de células normais através da ação dos receptores retinóides nucleares (RARs e RXRs) que são os principais mediadores destas ações que ao sofrerem alterações na sua expressão podem levar ao desenvolvimento e manutenção de tumores. No estudo da carcinogênese o modelo PDX é uma importante ferramenta pois mantém a arquitetura e microambiente do tumor original melhorando a compreensão de algumas vias, entre estas o processo de EMT/MET, na diferenciação das células tronco tumorais e quais receptores nucleares podem estar influenciando nestas vias. Objetivos: Analisar os padrões de comportamento biológico - tempo de formação e expansão do tumor e a manutenção dos padrões histológicos e de arquitetura do tumor original - em F0 e F1 no modelo PDX (xenoenxerto derivado de paciente) das amostras de centro de tumor e epitélio adjacente em camundongos Balb C/nude e avaliar a expressão gênica dos receptores retinóides, ALDH1 e marcadores do processo de EMT/MET por RT-PCR em PDX de carcinoma espinocelular oral em comparação com a amostra dos pacientes doadores nas passagens F(0) e F(1). Método: 24 camundongos Balb C/Nude, divididos em 2 grupos TG(I) – tumor graft paciente (I) e TG(II) – tumor graft paciente (II), subdivididos em 4 grupos de 3 animais: (A) – receberam PDX do centro do tumor; (B) – receberam PDX de epitélio adjacente ao tumor (margem de segurança cirúrgica); (C) receberam PDX de um animal do grupo (A); (D) receberam PDX de um animal do grupo (B). E Após estas fases, as amostras coletadas serão avaliadas por RT-PCR para comparação das expressões gênicas entre a amostra original (CT e EA) com os PDX´s nas passagens F(0) e F(1). Resultados: formação de tumores em todos os grupos – tanto do PDX de fragmento de centro do tumor quanto do PDX do epitélio adjacente. E A expressão gênica dos parâmetros observados não diferem no tumor original e passagem F(0) significativamente diferentes em F(1) (p<0,05). Conclusões: A técnida do PDX para o CEC é possível de ser realizada em menor tempo com a implantação de apenas um fragmento do tumor. Os tumores resultantes do PDX apresentaram tamanho suficiente para novas passagens, bem como para seu 6 uso em estudos de comportamento biológico das células neoplásicas. Quanto ao epitélio adjacente ao tumor (margem de segurança cirúrgica) constatou-se a presença de células tumorais com potencial de promover o crescimento de tumores devendo portanto ser melhor observada nas ressecções. O PDX de primeira passagem F(0) é o que mais se assemelha com o tumor original sendo o melhor para testes terapêuticos e estudos da carcinogênese do CEC oral. Keywords: CECP, modelo PDX, xenoenxerto, margem de segurança cirúrgica, , receptores retinóides, microdissecção a laser. / Introduction: Squamous cell carcinoma (SCC) represents 7% of all new cases of cancer in the world, with squamous cell carcinoma being the most frequent type. Both the biological behavior and the growth of the patients should be better understood, since the patients' survival show unobtrusive improvement in the last decades. PDX models were developed to study a tumor biology and especially the mechanisms of growth and proliferation through maintenance of the architecture and tumor microenvironment of the original tumor. Retinoids have a capacity to restore normal cell growth and differentiation through the action of nuclear retinoid receptors (RARs and RXRs) that are the main mediators and maintenance actions of tumors. In the study of carcinogenesis, the PDX model is an important tool because it maintains an architecture and microenvironment of the original tumor, improving an understanding of some pathways, among them in the EMT / MET process, the difference in tumor stem cells and which nuclear receptors may be influencing these routes. Objectives: To analyze changes in methodology and patterns of biological behavior - time of tumor formation and expansion and maintenance of histological and architectural patterns of the original tumor - in F0 and F1 without PDX model (patient derived xenograft) tumor and adjacent epithelium in Balb C / nude mice and to evaluate the gene expression of retinoid receptors, ALDH1 and EMT / MET process markers by RT-PCR in PDX of oral squamous cell carcinoma compared to a sample of donor patients in F ( 0) and F (1). Method: 24 Balb C / Nude mice, divided into 2 groups TG (I) - patient tumor graft (I) and TG (II) - patient tumor graft (II) subdivided into 4 groups of 3 animals: (A) - received PDX from the center of the tumor; (B) - received epithelial PDX adjacent to the tumor (surgical margin of safety); (C) received PDX from one animal of group (A); (D) received PDX from one animal of group (B). E After these phases, as samples collected for RT-PCR evaluation for comparison of gene expressions between an original sample (CT and EA) with F passages of PDX F (0) and F (1). Results: tumor formation in all groups - both the PDX of the tumor center fragment and the PDX of the adjacent epithelium. E The gene expression of the observed parameters did not differ without original tumor and F (0) differential passage in F (1) (p <0.05). Conclusions: The PDX technique for CPB is possible to be performed in a shorter time with a tumor fragment implantation. Tumors resulting from PDX presented the solution for new passages, as well as for their use in studies of the biological behavior of neoplastic cells. As for the epithelium adjacent to the tumor (surgical margin of safety), a presence of tumor cells with the potential to promote the growth of tumors has been observed and should therefore be better observed in the resections. The first pass PDX F (0) is the one that most closely resembles the 8 original tumor being the best for therapeutic tests and studies of oral SCC carcinogenesis.

Introdução: O carcinoma espinocelular (CEC) representa 7% de todos os novos casos de câncer no mundo, sendo o carcinoma espinocelular o tipo mais frequente. Tanto o comportamento biológico quanto o crescimento dos tumores devem ser melhores entendidos, uma vez que a sobrevida dos pacientes apresentou discreta melhora nas últimas décadas. Os modelos PDX foram desenvolvidos para estudar a biologia tumoral e principalmente os mecanismos de crescimento e proliferação através da manutenção da arquitetura e microambiente tumoral do tumor original. Os retinóides possuem a capacidade de restaurar o crescimento e a diferenciação de células normais através da ação dos receptores retinóides nucleares (RARs e RXRs) que são os principais mediadores destas ações que ao sofrerem alterações na sua expressão podem levar ao desenvolvimento e manutenção de tumores. No estudo da carcinogênese o modelo PDX é uma importante ferramenta pois mantém a arquitetura e microambiente do tumor original melhorando a compreensão de algumas vias, entre estas o processo de EMT/MET, na diferenciação das células tronco tumorais e quais receptores nucleares podem estar influenciando nestas vias. Objetivos: Analisar os padrões de comportamento biológico - tempo de formação e expansão do tumor e a manutenção dos padrões histológicos e de arquitetura do tumor original - em F0 e F1 no modelo PDX (xenoenxerto derivado de paciente) das amostras de centro de tumor e epitélio adjacente em camundongos Balb C/nude e avaliar a expressão gênica dos receptores retinóides, ALDH1 e marcadores do processo de EMT/MET por RT-PCR em PDX de carcinoma espinocelular oral em comparação com a amostra dos pacientes doadores nas passagens F(0) e F(1). Método: 24 camundongos Balb C/Nude, divididos em 2 grupos TG(I) – tumor graft paciente (I) e TG(II) – tumor graft paciente (II), subdivididos em 4 grupos de 3 animais: (A) – receberam PDX do centro do tumor; (B) – receberam PDX de epitélio adjacente ao tumor (margem de segurança cirúrgica); (C) receberam PDX de um animal do grupo (A); (D) receberam PDX de um animal do grupo (B). E Após estas fases, as amostras coletadas serão avaliadas por RT-PCR para comparação das expressões gênicas entre a amostra original (CT e EA) com os PDX´s nas passagens F(0) e F(1). Resultados: formação de tumores em todos os grupos – tanto do PDX de fragmento de centro do tumor quanto do PDX do epitélio adjacente. E A expressão gênica dos parâmetros observados não diferem no tumor original e passagem F(0) significativamente diferentes em F(1) (p<0,05). Conclusões: A técnida do PDX para o CEC é possível de ser realizada em menor tempo com a implantação de apenas um fragmento do tumor. Os tumores resultantes do PDX apresentaram tamanho suficiente para novas passagens, bem como para seu 6 uso em estudos de comportamento biológico das células neoplásicas. Quanto ao epitélio adjacente ao tumor (margem de segurança cirúrgica) constatou-se a presença de células tumorais com potencial de promover o crescimento de tumores devendo portanto ser melhor observada nas ressecções. O PDX de primeira passagem F(0) é o que mais se assemelha com o tumor original sendo o melhor para testes terapêuticos e estudos da carcinogênese do CEC oral. Keywords: CECP, modelo PDX, xenoenxerto, margem de segurança cirúrgica, , receptores retinóides, microdissecção a laser. / Introduction: Squamous cell carcinoma (SCC) represents 7% of all new cases of cancer in the world, with squamous cell carcinoma being the most frequent type. Both the biological behavior and the growth of the patients should be better understood, since the patients' survival show unobtrusive improvement in the last decades. PDX models were developed to study a tumor biology and especially the mechanisms of growth and proliferation through maintenance of the architecture and tumor microenvironment of the original tumor. Retinoids have a capacity to restore normal cell growth and differentiation through the action of nuclear retinoid receptors (RARs and RXRs) that are the main mediators and maintenance actions of tumors. In the study of carcinogenesis, the PDX model is an important tool because it maintains an architecture and microenvironment of the original tumor, improving an understanding of some pathways, among them in the EMT / MET process, the difference in tumor stem cells and which nuclear receptors may be influencing these routes. Objectives: To analyze changes in methodology and patterns of biological behavior - time of tumor formation and expansion and maintenance of histological and architectural patterns of the original tumor - in F0 and F1 without PDX model (patient derived xenograft) tumor and adjacent epithelium in Balb C / nude mice and to evaluate the gene expression of retinoid receptors, ALDH1 and EMT / MET process markers by RT-PCR in PDX of oral squamous cell carcinoma compared to a sample of donor patients in F ( 0) and F (1). Method: 24 Balb C / Nude mice, divided into 2 groups TG (I) - patient tumor graft (I) and TG (II) - patient tumor graft (II) subdivided into 4 groups of 3 animals: (A) - received PDX from the center of the tumor; (B) - received epithelial PDX adjacent to the tumor (surgical margin of safety); (C) received PDX from one animal of group (A); (D) received PDX from one animal of group (B). E After these phases, as samples collected for RT-PCR evaluation for comparison of gene expressions between an original sample (CT and EA) with F passages of PDX F (0) and F (1). Results: tumor formation in all groups - both the PDX of the tumor center fragment and the PDX of the adjacent epithelium. E The gene expression of the observed parameters did not differ without original tumor and F (0) differential passage in F (1) (p <0.05). Conclusions: The PDX technique for CPB is possible to be performed in a shorter time with a tumor fragment implantation. Tumors resulting from PDX presented the solution for new passages, as well as for their use in studies of the biological behavior of neoplastic cells. As for the epithelium adjacent to the tumor (surgical margin of safety), a presence of tumor cells with the potential to promote the growth of tumors has been observed and should therefore be better observed in the resections. The first pass PDX F (0) is the one that most closely resembles the 8 original tumor being the best for therapeutic tests and studies of oral SCC carcinogenesis.

Introdução: O carcinoma espinocelular (CEC) representa 7% de todos os novos casos de câncer no mundo, sendo o carcinoma espinocelular o tipo mais frequente. Tanto o comportamento biológico quanto o crescimento dos tumores devem ser melhores entendidos, uma vez que a sobrevida dos pacientes apresentou discreta melhora nas últimas décadas. Os modelos PDX foram desenvolvidos para estudar a biologia tumoral e principalmente os mecanismos de crescimento e proliferação através da manutenção da arquitetura e microambiente tumoral do tumor original. Os retinóides possuem a capacidade de restaurar o crescimento e a diferenciação de células normais através da ação dos receptores retinóides nucleares (RARs e RXRs) que são os principais mediadores destas ações que ao sofrerem alterações na sua expressão podem levar ao desenvolvimento e manutenção de tumores. No estudo da carcinogênese o modelo PDX é uma importante ferramenta pois mantém a arquitetura e microambiente do tumor original melhorando a compreensão de algumas vias, entre estas o processo de EMT/MET, na diferenciação das células tronco tumorais e quais receptores nucleares podem estar influenciando nestas vias. Objetivos: Analisar os padrões de comportamento biológico - tempo de formação e expansão do tumor e a manutenção dos padrões histológicos e de arquitetura do tumor original - em F0 e F1 no modelo PDX (xenoenxerto derivado de paciente) das amostras de centro de tumor e epitélio adjacente em camundongos Balb C/nude e avaliar a expressão gênica dos receptores retinóides, ALDH1 e marcadores do processo de EMT/MET por RT-PCR em PDX de carcinoma espinocelular oral em comparação com a amostra dos pacientes doadores nas passagens F(0) e F(1). Método: 24 camundongos Balb C/Nude, divididos em 2 grupos TG(I) – tumor graft paciente (I) e TG(II) – tumor graft paciente (II), subdivididos em 4 grupos de 3 animais: (A) – receberam PDX do centro do tumor; (B) – receberam PDX de epitélio adjacente ao tumor (margem de segurança cirúrgica); (C) receberam PDX de um animal do grupo (A); (D) receberam PDX de um animal do grupo (B). E Após estas fases, as amostras coletadas serão avaliadas por RT-PCR para comparação das expressões gênicas entre a amostra original (CT e EA) com os PDX´s nas passagens F(0) e F(1). Resultados: formação de tumores em todos os grupos – tanto do PDX de fragmento de centro do tumor quanto do PDX do epitélio adjacente. E A expressão gênica dos parâmetros observados não diferem no tumor original e passagem F(0) significativamente diferentes em F(1) (p<0,05). Conclusões: A técnida do PDX para o CEC é possível de ser realizada em menor tempo com a implantação de apenas um fragmento do tumor. Os tumores resultantes do PDX apresentaram tamanho suficiente para novas passagens, bem como para seu 6 uso em estudos de comportamento biológico das células neoplásicas. Quanto ao epitélio adjacente ao tumor (margem de segurança cirúrgica) constatou-se a presença de células tumorais com potencial de promover o crescimento de tumores devendo portanto ser melhor observada nas ressecções. O PDX de primeira passagem F(0) é o que mais se assemelha com o tumor original sendo o melhor para testes terapêuticos e estudos da carcinogênese do CEC oral. Keywords: CECP, modelo PDX, xenoenxerto, margem de segurança cirúrgica, , receptores retinóides, microdissecção a laser. / Introduction: Squamous cell carcinoma (SCC) represents 7% of all new cases of cancer in the world, with squamous cell carcinoma being the most frequent type. Both the biological behavior and the growth of the patients should be better understood, since the patients' survival show unobtrusive improvement in the last decades. PDX models were developed to study a tumor biology and especially the mechanisms of growth and proliferation through maintenance of the architecture and tumor microenvironment of the original tumor. Retinoids have a capacity to restore normal cell growth and differentiation through the action of nuclear retinoid receptors (RARs and RXRs) that are the main mediators and maintenance actions of tumors. In the study of carcinogenesis, the PDX model is an important tool because it maintains an architecture and microenvironment of the original tumor, improving an understanding of some pathways, among them in the EMT / MET process, the difference in tumor stem cells and which nuclear receptors may be influencing these routes. Objectives: To analyze changes in methodology and patterns of biological behavior - time of tumor formation and expansion and maintenance of histological and architectural patterns of the original tumor - in F0 and F1 without PDX model (patient derived xenograft) tumor and adjacent epithelium in Balb C / nude mice and to evaluate the gene expression of retinoid receptors, ALDH1 and EMT / MET process markers by RT-PCR in PDX of oral squamous cell carcinoma compared to a sample of donor patients in F ( 0) and F (1). Method: 24 Balb C / Nude mice, divided into 2 groups TG (I) - patient tumor graft (I) and TG (II) - patient tumor graft (II) subdivided into 4 groups of 3 animals: (A) - received PDX from the center of the tumor; (B) - received epithelial PDX adjacent to the tumor (surgical margin of safety); (C) received PDX from one animal of group (A); (D) received PDX from one animal of group (B). E After these phases, as samples collected for RT-PCR evaluation for comparison of gene expressions between an original sample (CT and EA) with F passages of PDX F (0) and F (1). Results: tumor formation in all groups - both the PDX of the tumor center fragment and the PDX of the adjacent epithelium. E The gene expression of the observed parameters did not differ without original tumor and F (0) differential passage in F (1) (p <0.05). Conclusions: The PDX technique for CPB is possible to be performed in a shorter time with a tumor fragment implantation. Tumors resulting from PDX presented the solution for new passages, as well as for their use in studies of the biological behavior of neoplastic cells. As for the epithelium adjacent to the tumor (surgical margin of safety), a presence of tumor cells with the potential to promote the growth of tumors has been observed and should therefore be better observed in the resections. The first pass PDX F (0) is the one that most closely resembles the 8 original tumor being the best for therapeutic tests and studies of oral SCC carcinogenesis.

INTRODUÇÃO: O mixofibrossarcoma (MFS) é um dos mais frequentes sarcomas de partes moles (SPM) em idosos que afeta principalmente as extremidades. Historicamente, é um grupo de tumores heterogêneos. Clinicamente está caracterizado por apresentar uma alta incidência de recorrência local (RL) e um conhecimento limitado sobre a sua capacidade de metástase. O índice de RL após a ressecção cirúrgica é relativamente maior quando comparado com outros SPM. Não existe um consenso em como identificar os pacientes com maior risco. O objetivo deste estudo foi analisar os fatores de prognóstico dos pacientes diagnosticados com MFS em uma instituição única. MÉTODOS: Foram analisados retrospectivamente os prontuários de 75 pacientes com diagnóstico confirmado de MFS nas extremidades, que foram submetidos a tratamento cirúrgico, nos últimos 25 anos. Comparamos idade, sexo, tamanho e localização do tumor, grau histológico segundo a Fédération Nationale des Centres de Lutte Contre le Cancer (FNCLCC) e o estádio segundo a American Joint Committee on Cancer (AJCC). A média de idade foi 49,7 anos. A localização foi: membro superior (25,4%), membro inferior (66,6%) e pelve (8%). Os pacientes apresentaram baixo, intermediário e alto grau, em: 29,3%, 24% e 46,7% dos tumores, respectivamente. Pelo tamanho foram categorizados em: =/< 5 cm (13,3%), > 5 e <10 cm (42,7%), > 10 e <15 cm (9%) e =/> 15 cm (18,7%). Em total, 26,7% receberam radioterapia pós-operatória. As margens foram livres em 76% e comprometidas em 24%. A análise da regressão de Cox bivariada foi utilizada para determinar as associações entre os fatores clínicos e de tratamento com a RL. RESULTADOS: O tempo médio de seguimento foi 30.7 meses. O 26,7% dos pacientes apresentaram RL. Metástase foi reportada em 27 (36%) pacientes. O local mais comum de metástase foi: pulmão (92,6%) e gânglios linfáticos (18,5%). O tempo médio de sobrevida dos pacientes com metástase foi 21,2 meses. Os fatores preditivos para RL foram: margens comprometidas (hazard ratio 5.47, 95% intervalo de confiança, 2.23-13.40, P < 0.001) e metástase (hazard ratio 10.24, 95% intervalo de confiança, 3.53-26.68, P < 0.001). Os fatores preditivos da sobrevida livre de RL foram: grau histológico, margens comprometidas (hazard ratio 3.18, 95% intervalo de confiança, 1.51-6.70, P =0.001), e metástase (hazard ratio 15.23, 95% intervalo de confiança, 5.57-41.61, P= 0.001). Os fatores preditivos de sobrevida em geral foram: RL (hazard ratio 5.13, 95% intervalo de confiança, 2.15-12.24, P < 0.001), e metástase (hazard ratio 540.97, 95% intervalo de confiança, 5.04-58112.03, P < 0.001). CONCLUSÃO: As margens cirúrgicas comprometidas e a metástase estão diretamente associadas com a RL. O grau histológico do tumor, as margens comprometidas, a RL e a metástase, são fatores de pior prognóstico no MFS / BACKGROUND AND AIMS: Myxofibrosarcoma (MFS) is one of the most common soft tissue sarcomas (STS) in elderly patients and it primarily affects the extremities. They are a historically heterogeneous group of tumors. The clinical course of MFS is characterized by a high incidence of local recurrences (LR), but knowledge about distant metastasis is sparse. MFS is reported to have a higher risk of LR following definitive surgical excision relative to other STS. There is no agreement on how to identify patients at major risk. The objectives of this study were to analyze the prognostic factors and outcomes of patients with MFS treated at a single institution. METHODS: We retrospectively reviewed the records of 75 patients with pathologically confirmed MFS of the extremities who underwent surgery in the last 25 years. We compared the age, sex, tumor size and location, Fédération Nationale des Centres de Lutte Contre le Cancer (FNCLCC) grade and the American Joint Committee on Cancer (AJCC) stage. Median age was 49.7 years (range, 1 to 88 y). Site of disease was: upper extremity (25.4%), lower extremity (66.6%) and pelvic (8%). Patients had low, intermediate and high-grade, in: 29.3%, 24% and 46.7% of tumors, respectively. Tumors were categorized as =/< 5 cm (13.3%), > 5 and < 10 cm (42.7%), > 10 and < 15 cm (9%) and =/> 15 cm (18.7%). In total, 26.7% received postoperative radiotherapy. All patients underwent surgery. Margins were negative in 76% and positive in 24%. Bivariate Cox regression analysis was utilized to determine associations between clinical and treatment factors with LR. RESULTS: Median follow-up time was 30.7 months (range, 1.8 to 383.8 m). We found a 26.7% of LR. Distant metastasis was reported in 27 (36%) patients. The most common sites of metastasis were: lung (92.6%) and lymph nodes (18.5%). The overall survival rate in patients with metastasis was 21.2 months (range, 4.8 to 114.8 m). Predictors of LR were: positive margins (hazard ratio 5.47, 95% confidence interval, 2.23-13.40, P < 0.001) and distant metastasis (hazard ratio 10.24, 95% confidence interval, 3.53-26.68, P < 0.001). Predictors of overall survival free of LR were: grade, positive margins (hazard ratio 3.18, 95% confidence interval, 1.51-6.70, P =0.001), and distant metastasis (hazard ratio 15.23, 95% confidence interval, 5.57-41.61, P= 0.001). Predictors of overall survival were grade, LR (hazard ratio 5.13, 95% confidence interval, 2.15-12.24, P < 0.001), and distant metastasis (hazard ratio 540.97, 95% confidence interval, 5.04-58112.03, P < 0.001). CONCLUSION: In this institutional series of MFS, positive margins and distant metastasis were significantly associated with a higher risk of LR. Tumor grade, LR, positive margins and distant metastases were significant predictors of overall survival poor prognosis

<div> Advancements in cancer treatments have increased rapidly in recent years, but cures remain elusive. Surgical tumor resection is a central treatment for many solid malignancies. Residual tumor at surgical margins leads to tumor recurrence. Novel tools for assessing residual tumor at surgical margins could improve surgical outcomes by helping to maximize the extent of resection. Ambient ionization-mass spectrometry (MS) methods generate and analyze ions from minimally prepared samples in near-real-time (e.g. seconds to minutes). These methods leverage the high sensitivity and specificity of mass spectrometry for analyzing gas phase ions and generating those ions quickly and with minimal sample preparation. Recent work has shown that differential profiles of ions, corresponding to phospholipids and small metabolites, are detected from cancerous and their respective normal tissue with ambient ionization-MS methods. When properly implemented, ambient ionization-MS could be used to assess for tumor at surgical margins and provide a molecular diagnosis during surgery. </div><div><br></div><div>The research herein reports efforts in developing rapid intraoperative ambient ionization-MS methods for the molecular assessment of cancerous tissues. Touch spray (TS) ionization and desorption electrospray ionization (DESI) were utilized to analyze kidney cancer and brain cancer.</div><div><br></div><div> As a demonstration of the applicability of TS-MS to provide diagnostic information from fresh surgical tissues, TS-MS was used to rapidly analyze renal cell carcinoma and healthy renal tissue biopsies obtained from human subjects undergoing nephrectomy surgery. Differential phospholipid profiles were identified using principal component analysis (PCA), and the significant ions were characterized using multiple stages of mass spectrometry and high resolution/exact mass MS. The same TS-MS analyzed renal tissues were subsequently analyzed with DESI-MS imaging to corroborate the TS-MS results, and the significant DESI-MS ions were also characterized with MS.</div><div><br></div><div>Significant efforts were made in developing and evaluating a standalone intraoperative DESI-MS system for analyzing brain tissue biopsies during brain tumor surgery. The intraoperative DESI-MS system consists of a linear trap quadrupole mass spectrometer placed on a custom-machined cart that contains all hardware for operating the mass spectrometer. This instrument was operated in the neurosurgical suites at Indiana University School of Medicine to rapidly analyze brain tissue biopsies obtained from glioma resection surgeries. A DESI-MS library of normal brain tissue and glioma was used to statistically classify the brain tissue biopsies collected in the operating room. Multivariate statistical methodologies were employed to predict the disease state and tumor cell percentage of the samples. A DESI-MS assay for detecting 2-hydroxyglutarate (2HG), the oncometabolic product of the isocitrate dehydrogenase (IDH) mutation (a key glioma prognostic marker), was developed and applied to determine the IDH mutation status during the surgical resection. The strengths, weaknesses, and areas of future work in this field are discussed. </div><div><br></div>

<p>My research has focused on the development and translation of ambient ionization mass spectrometry (MS)-based platforms in clinical and surgical settings, specifically in the area of brain cancer diagnostics and surgical decision making. Ambient ionization MS methods, such as those described herein, generate and analyze gas phase ions with high sensitivity and specificity from minimally prepared samples in near-real-time, on the order of seconds to minutes, rendering them well suited to point-of-care applications. We used ambient ionization MS methods, specifically desorption electrospray ionization mass spectrometry (DESI-MS) and extraction nanoelectrospray ionization mass spectrometry (nESI-MS) to molecularly characterize brain cancer biopsies. The characterization was made using diagnostic compounds identified as markers of disease state, tissue composition, tumor type, and genotype in human brain tissue. Methods were developed and validated offline in the laboratory and translated to clinical and surgical settings, thereby generating chemical information on prognostic features intraoperatively and providing valuable information that would be otherwise unavailable. We believe that, with approval, the methodologies described can assist physicians and improve patient outcomes by providing analytical tools and molecular information that can inform surgical decision making and adjuvant treatment strategies, complementing and not interfering with standard of care protocols.</p> <p><br></p> <p>We have successfully demonstrated the use of desorption electrospray ionization mass spectrometry (DESI-MS) for the expedient molecular assessment of human glioma tissue biopsies based on lipid profiles and prognostic metabolites, both at the tumor core and near surgical margins, in two small-scale, clinical studies. Maximal surgical resection of gliomas that avoids non-infiltrated tissue is associated with survival benefit in patients with glioma. The infiltrative nature of gliomas, as well as their morphological and genetic diversity, renders treatment difficult and demands an integrated imaging and diagnostic approach during surgery to guide clinicians in achieving maximal tumor resection. Further, the estimation of tumor cell percentage (TCP), a measure of tumor infiltration at surgical margins, is not routinely assessed intraoperatively. </p> <p>We have previously shown that rapid, offline molecular assessment of tumor infiltration in tissue biopsies is possible and believe that the same assessment performed intraoperatively in biopsied tissue near surgical margins could improve resection and better inform patient management strategies, including postoperative radiotherapy. Using a DESI-MS spectral library of normal brain tissue and glioma biopsies to generate a statistical model to classify brain tissue biopsies intraoperatively, multivariate statistical approaches were used to predict the disease state and tumor cell percentage (TCP) of each biopsy, thereby providing an measure of tumor infiltration at surgical margins via molecular indicators. In addition to assessment of tumor infiltration, we have developed DESI-MS assays for detecting the oncometabolite 2-Hydroxyglutarate (2HG) to detect isocitrate dehydrogenase (IDH) mutations in gliomas intraoperatively. Knowledge of IDH genotypes at the time of surgical resection could improve patient outcomes, as more aggressive tumor resection of IDH-mutated gliomas is associated with increased survival. While assessments of IDH genotype are typically not available until days after surgery, we have demonstrated the ability to provide this information is less than five minutes. An intraoperative DESI-MS system has successfully been used in a proof-of-concept clinical study and intraoperative performance validation of this platform is ongoing. The findings of these two studies as well as strengths, weaknesses, and areas of improvement for upcoming future iterations of the research are discussed.</p> <p><br></p> <p>Point-of-care applications necessitate the adaptation of MS methodologies to smaller devices. Miniature mass spectrometers (Mini MS) boast small footprints, simple operation, and low power consumption, noise levels, and cost, making them attractive candidates for point-of-care use. In a small-scale clinical study, we demonstrated the first application of a Mini MS for determination of IDH mutation status in gliomas intraoperatively. This study paves a path forward for the application of Mini MS in the OR. With its small footprint and low power consumption and noise level, this application of miniature mass spectrometers represents a simple and cost-effective platform for an important intraoperative measurement. </p> <p><br></p> <p>While MS-based methods of tissue analysis can detect molecular features of interest and rapidly produce large quantities of data, their inherent speed is rarely utilized because they are traditionally coupled with time-consuming separation techniques (e.g., chromatography). Ambient ionization MS, specifically DESI-MS, is well suited for high-throughput applications due to its lack of sample preparation and purification techniques. In an attempt to rapidly characterize microarrays of tissue biopsies, we developed a high-throughput DESI-MS (HT-DESI-MS) method for the rapid characterization of disease state, human brain tumor type, glioma classification, and detection of IDH mutations in tissue microarrays (TMA) of banked and fresh human brain tissue biopsies. We anticipate that HT-DESI-MS analysis of TMAs could become a standard tool for the generation of spectral libraries for sample classification, the identification of biomarkers through large-scale studies, the correlation of molecular features with anatomical features when coupled to digital pathology, and the assessment of drug efficacy. </p>