Gene expression in B-1 cells from lupus-prone mice.

New Zealand Black X New Zealand White F1 [(NZB/NZW)F1] mice develop an autoimmune condition with similarities to human systemic lupus erythematosus (SLE). In this study, we demonstrate that B-1 cells, which have previously been reported to be involved in several autoimmune diseases, have altered gene expression in these mice. RNA was extracted from purified B-1 cells of disease-free C57BL/6 mice and lupus-prone (NZB/NZW)F1 mice. Gene expression was analysed using DNA microarray techniques and validated by real time reverse transcriptase polymerase chain reaction (RT-PCR). In (NZB/NZW)F1 mice, some genes had altered expression patterns compared to disease-free controls. Specifically, the upregulation of Ifitm1, Pvrl2 and Ifi202b and downregulation of Trp53bp1 mRNA were observed in (NZB/NZW)F1 mice. These genes are known to be associated with autoimmune diseases. This pattern of gene expression in B-1 cells could understanding of the pathogenesis of SLE. Thus, it is reasonable to hypothesise that the altered gene expression observed in B-1 cells in our experimental model is important for SLE prognosis and therapy, and these implications are discussed herein.

Exposure to arsenic at levels found inU.S. drinking water modifies expression in the mouse lung.

The mechanisms of action of drinking water arsenic in the lung and the threshold for biologic effects remain controversial. Our study utilizes Affymetrix 22,690 transcript oligonucleotide microarrays to assess the long-term effects of increasing doses of drinking water arsenic on expression levels in the mouse lung. Mice were exposed at levels commonly found in contaminated drinking water wells in the United States (0, 0.1, 1 ppb), as well as the 50 ppb former maximum contaminant level, for 5 weeks. The expression profiles revealed modification of a number of important signaling pathways, many with corroborating evidence of arsenic responsiveness. We observed statistically significant expression changes for transcripts involved in angiogenesis, lipid metabolism, oxygen transport, apoptosis, cell cycle, and immune response. Validation by reverse transcription-PCR and immunoblot assays confirmed expression changes for a subset of transcripts. These data identify arsenic-modified signaling pathways that will help guide investigations into mechanisms of arsenic's health effects and clarify the threshold for biologic effects and potential disease risk.

Dorsal capsulodesis associated with arthoscopy-assisted scapholunate ligament reconstruction using a palmaris longus tendon graft.

OBJECTIVES: To measure the quality of life, the time to work return, and clinical, functional, and radiographic parameters of patients treated with dorsal capsulodesis associated with scapholunate (SL) reconstruction, assisted by arthroscopy. METHODS: From January 2015 to September 2016, 14 adult patients with SL dissociation underwent surgical treatment with the SL reconstruction procedure assisted by arthroscopy, using the new technique proposed in this study. All patients were assessed by the occupational therapy department at regular intervals after surgery and performed the same sequence of rehabilitation. The parameters analyzed were: range of motion (ROM), Disability of the Arm, Shoulder, and Hand (DASH), visual analog scale (VAS), and radiographic analysis to visualize the pre- and postoperative SL gap and the pre- and postoperative dorsal intercalated segment instability (DISI) deformity the. The complications and the time to return to work activities were described. RESULTS: The follow-up time was 12 months (3-17). The ROM averaged 321° (96.9% of the normal side). VAS was 1.79/10 (1-6). DASH was 6.50/100 (1-30). The time to work return work was 4.42 months (2-17). As for complications, one patient developed SLAC, and underwent four-corner fusion one year after ligament reconstruction. Currently, he has experienced pain relief, with a functional range of motion of the wrist, and has not yet returned to professional activities.The preoperative SL gap was 4.29 mm (2-7); in the postoperative period, it was 1.79 mm (1-4). The DISI deformity was present in ten patients with SL angle > 70° (preoperative) and it was corrected after surgery, in all patients. SLAC stage I was identified in a patient. Arthroscopy was performed in all cases. The SL instability was classified as Geissler grade III in four cases and as grade IV in ten cases. CONCLUSION: The new approach (dorsal capsulodesis associated with SL reconstruction, assisted by arthroscopy) presented in this study is safe and effective in the treatment of SL dissociation, since it offers satisfactory clinical, radiographic and functional results, showing low rates of complications. For patients, it allows the return to their social and professional activities, and increases their life quality.

OBJETIVOS: Mensurar a qualidade de vida, o tempo de retorno ao trabalho, os resultados clínicos, funcionais e radiográficos dos pacientes submetidos à capsulodese dorsal associada à reconstrução ligamentar escafossemilunar assistida por artroscopia. MÉTODOS: De janeiro de 2015 a setembro de 2016, 14 pacientes, esqueleticamente maduros, adultos, com dissociação escafolunar (SL), foram submetidos ao tratamento cirúrgico com o procedimento de reconstrução do ligamento escafossemilunar assistido por artroscopia com a nova técnica proposta neste estudo. Todos os pacientes foram avaliados pelo setor de terapia ocupacional em intervalos regulares de pós-operatório e fizeram a mesma sequência de reabilitação. Os parâmetros analisados foram: arco de movimento (ADM), Disability Arm, Shoulder and Hand (Dash), escala visual analógica (EVA) e análise radiográfica pré e pós-operatória para visualizar o espaço escafolunar (sinal de Terry-Thomas) e deformidade em Dorsal Intercalated Segment Instability (DISI) pré e pós-operatória. Descrição das complicações e o tempo de retorno ao trabalho. RESULTADOS: O tempo de seguimento foi de 12 meses [3-17]. O ADM foi em média 321,07° (96,9% do lado normal). O valor da avaliação subjetiva da dor (VAS) foi 1,79/10 [1-6]. A mensuração da qualidade de vida pelo Dash foi de 6,50/100 [1-30]. O tempo de retorno ao trabalho foi de 4,42 meses [2-17]. Quanto às complicações, uma paciente evoluiu com SLAC e foi submetida à artrodese dos quatro cantos um ano após a reconstrução ligamentar. Evoluiu com melhoria da dor e está com o ADM do punho funcional, mas ainda não retornou às atividades profissionais. O intervalo do SL (gap) pré-operatório foi de 4,29 mm [2-7] e o pós-operatório foi de 1,79 mm [1-4]. A deformidade DISI estava presente em dez pacientes, com um ângulo SL acima de 70° (pré-operatório), e foi corrigida após a cirurgia em todos os pacientes. SLAC estágio I foi identificado em um paciente. A artroscopia foi feita em todos os casos. A instabilidade SL foi classificada como um grau Geissler III em quatro casos e grau IV em dez casos. CONCLUSÃO: A nova abordagem (capsulodese dorsal associada à reconstrução ligamentar escafossemilunar assistida por artroscopia) apresentada neste estudo é segura e eficaz no tratamento da dissociação escafolunar, já que apresenta resultados radiográficos, clínicos e funcionais satisfatórios, demonstra baixas taxas de complicações, permite o retorno às atividades sociais e profissionais e aumenta a qualidade de vida desses pacientes.

Hope matters to the glycemic control of adolescents and young adults with type 1 diabetes.

This study investigated the association of hope and its factors with depression and glycemic control in adolescents and young adults with type 1 diabetes. A total of 113 patients were invited to participate. Significant negative correlations were found between hope and HbA1c and also between hope and depression. Hope showed a significant association with HbA1c and depression in the stepwise regression model. Among the hope factors, "inner positive expectancy" was significantly associated with HbA1c and depression. This study supports that hope matters to glycemic control and depression. Intervention strategies focusing on hope should be further explored.

Web-based learning in undergraduate medical education: development and assessment of an online course on experimental surgery.

In order to increase the number of practical and discussion classes offered to students in the traditional-curriculum scenario, while decreasing the lecture-based ones and to create an online community to share knowledge on surgery, we developed and assessed the first online course for undergraduate medical students on experimental surgery at the Federal University of Sao Paulo-UNIFESP, Brazil. The purposes of the present study are: describe and discuss the process and the lessons learned involved in developing an undergraduate web-based course and analyze the students' attitude towards this educational environment. A group of medical students was taught online during 5 weeks on the theory of experimental surgery through video quizzes, required readings, collaborative activities using discussion board and asynchronous communication. The students' knowledge gain, their web session variables and the results of the course evaluation were used to support our study. The students have significantly improved their knowledge on experimental surgery after the course. Among factors in the online course that could possibly have contributed to this gain, the interactive activities (video quizzes), key element in our online material, seemed to be promising for candidates. The evaluation results demonstrated high levels of course functionality, effectiveness of its online content and acceptance among medical students. This study indicated that a web-based course for undergraduate students may be successfully developed and implemented in medical settings and the students seem to be quite supportive. We encourage undergraduate medical learning strategies involving the Web.

The impact of knowledge about diabetes, resilience and depression on glycemic control: a cross-sectional study among adolescents and young adults with type 1 diabetes.

BACKGROUND: The purpose of this study is to evaluate the relationship between glycemic control and the factors of knowledge about diabetes, resilience, depression and anxiety among Brazilian adolescents and young adults with type 1 diabetes. METHODS: This cross-sectional study included 85 adolescents and young adults with type 1 diabetes, aged between 11-22 years, with an average age of 17.7 ± 3.72 years. Glycemic control degree was evaluated through HbA1c. To assess psychosocial factors, the following questionnaires were used: resilience (Resilience Scale, RS) and anxiety and depression (Hospital Anxiety and Depression Scale, HADS). The Diabetes Knowledge Assessment Scale (DKNA) was used to assess knowledge about diabetes. RESULTS: Significant correlations were found between HbA1c and resilience, anxiety and depression. Multiple linear regression analysis revealed that the only variable which presented significant association with the value of HbA1c was depression. CONCLUSIONS: Depression has a significant association with higher HbA1c levels, as demonstrated in a regression analysis. The results suggest that depression, anxiety and resilience should be considered in the design of a multidisciplinary approach to type 1 diabetes, as these factors were significantly correlated with glycemic control. Glycemic control was not correlated with knowledge of diabetes, suggesting that theoretical or practical understanding of this disease is not by itself significantly associated with appropriate glycemic control (HbA1c = 7.5%).

Crosstalk between B16 melanoma cells and B-1 lymphocytes induces global changes in tumor cell gene expression.

The analysis of gene expression patterns in cancers has improved the understanding of the mechanisms underlying the process of metastatic progression. However, the acquisition of invasive behavior in melanoma is poorly understood. In melanoma, components of the immune system can contribute to tumor progression, and inflammatory cells can influence almost all aspects of cancer progression, including metastasis. Recent studies have attributed an important role to B-1 cells, a subset of B lymphocytes, in melanoma progression. In vitro interactions between B16 melanoma cells and B-1 lymphocytes lead to increased B16 cell metastatic potential, but the molecular changes induced by B-1 lymphocytes on B16 cells have not yet been elucidated. In this study, we used a microarray approach to assess the gene expression profile of B16 melanoma cells following contact with B-1 lymphocytes (B16B1). The microarray analysis identified upregulation in genes involved with metastatic progression, such as ctss, ccl5, cxcl2 and stat3. RT-qPCR confirmed this increase in mRNA expression in B16B1 samples. As previous studies have indicated that the ERK1/2 MAPK cascade is activated in melanoma cells following contact with B-1 lymphocytes, RT-qPCR was performed with RNA from melanoma cells before and after contacting B-1 cells and untreated or treated with ERK phosphorylation inhibitors. The results showed that the expression of stat3, ctss and cxcl2 increased in B16B1 but decreased following ERK1/2 MAPK inhibition. Ccl5 gene expression increased after contacting B-1 cells and was maintained at the same level following inhibitor treatment. Stat3 was verified and validated at the protein level by Western blot analysis. STAT3 expression was also significantly increased in B16B1, suggesting that this pathway can also contribute to the increased metastatic phenotype observed in our model. These results indicated that B-1 cells induce important global gene expression changes in B16 melanoma cells. We also evaluated the relationship of some of the genes identified as differentially expressed and the ERK1/2 MAPK cascade. This work may have important implications for understanding the role of B-1 lymphocytes and the ERK/MAPK cascade in the metastatic process.

Dorsal capsulodesis associated with arthoscopy-assisted scapholunate ligament reconstruction using a palmaris longus tendon graft / Capsulodese dorsal associada à reconstrução assistida por artroscopia do ligamento escafossemilunar com enxerto do tendão do músculo palmar longo

ABSTRACT OBJECTIVES: To measure the quality of life, the time to work return, and clinical, functional, and radiographic parameters of patients treated with dorsal capsulodesis associated with scapholunate (SL) reconstruction, assisted by arthroscopy. METHODS: From January 2015 to September 2016, 14 adult patients with SL dissociation underwent surgical treatment with the SL reconstruction procedure assisted by arthroscopy, using the new technique proposed in this study. All patients were assessed by the occupational therapy department at regular intervals after surgery and performed the same sequence of rehabilitation. The parameters analyzed were: range of motion (ROM), Disability of the Arm, Shoulder, and Hand (DASH), visual analog scale (VAS), and radiographic analysis to visualize the pre- and postoperative SL gap and the pre- and postoperative dorsal intercalated segment instability (DISI) deformity the. The complications and the time to return to work activities were described. RESULTS: The follow-up time was 12 months (3-17). The ROM averaged 321° (96.9% of the normal side). VAS was 1.79/10 (1-6). DASH was 6.50/100 (1-30). The time to work return work was 4.42 months (2-17). As for complications, one patient developed SLAC, and underwent four-corner fusion one year after ligament reconstruction. Currently, he has experienced pain relief, with a functional range of motion of the wrist, and has not yet returned to professional activities. The preoperative SL gap was 4.29 mm (2-7); in the postoperative period, it was 1.79 mm (1-4). The DISI deformity was present in ten patients with SL angle > 70° (preoperative) and it was corrected after surgery, in all patients. SLAC stage I was identified in a patient. Arthroscopy was performed in all cases. The SL instability was classified as Geissler grade III in four cases and as grade IV in ten cases. CONCLUSION: The new approach (dorsal capsulodesis associated with SL reconstruction, assisted by arthroscopy) presented in this study is safe and effective in the treatment of SL dissociation, since it offers satisfactory clinical, radiographic and functional results, showing low rates of complications. For patients, it allows the return to their social and professional activities, and increases their life quality.

RESUMO OBJETIVOS: Mensurar a qualidade de vida, o tempo de retorno ao trabalho, os resultados clínicos, funcionais e radiográficos dos pacientes submetidos à capsulodese dorsal associada à reconstrução ligamentar escafossemilunar assistida por artroscopia. MÉTODOS: De janeiro de 2015 a setembro de 2016, 14 pacientes, esqueleticamente maduros, adultos, com dissociação escafolunar (SL), foram submetidos ao tratamento cirúrgico com o procedimento de reconstrução do ligamento escafossemilunar assistido por artroscopia com a nova técnica proposta neste estudo. Todos os pacientes foram avaliados pelo setor de terapia ocupacional em intervalos regulares de pós-operatório e fizeram a mesma sequência de reabilitação. Os parâmetros analisados foram: arco de movimento (ADM), Disability Arm, Shoulder and Hand (Dash), escala visual analógica (EVA) e análise radiográfica pré e pós-operatória para visualizar o espaço escafolunar (sinal de Terry-Thomas) e deformidade em Dorsal Intercalated Segment Instability (DISI) pré e pós-operatória. Descrição das complicações e o tempo de retorno ao trabalho. RESULTADOS: O tempo de seguimento foi de 12 meses [3-17]. O ADM foi em média 321,07° (96,9% do lado normal). O valor da avaliação subjetiva da dor (VAS) foi 1,79/10 [1-6]. A mensuração da qualidade de vida pelo Dash foi de 6,50/100 [1-30]. O tempo de retorno ao trabalho foi de 4,42 meses [2-17]. Quanto às complicações, uma paciente evoluiu com SLAC e foi submetida à artrodese dos quatro cantos um ano após a reconstrução ligamentar. Evoluiu com melhoria da dor e está com o ADM do punho funcional, mas ainda não retornou às atividades profissionais. O intervalo do SL (gap) pré-operatório foi de 4,29 mm [2-7] e o pós-operatório foi de 1,79 mm [1-4]. A deformidade DISI estava presente em dez pacientes, com um ângulo SL acima de 70° (pré-operatório), e foi corrigida após a cirurgia em todos os pacientes. SLAC estágio I foi identificado em um paciente. A artroscopia foi feita em todos os casos. A instabilidade SL foi classificada como um grau Geissler III em quatro casos e grau IV em dez casos. CONCLUSÃO: A nova abordagem (capsulodese dorsal associada à reconstrução ligamentar escafossemilunar assistida por artroscopia) apresentada neste estudo é segura e eficaz no tratamento da dissociação escafolunar, já que apresenta resultados radiográficos, clínicos e funcionais satisfatórios, demonstra baixas taxas de complicações, permite o retorno às atividades sociais e profissionais e aumenta a qualidade de vida desses pacientes.

Mining gene expression signature for the detection of pre-malignant melanocytes and early melanomas with risk for metastasis.

BACKGROUND: Metastatic melanoma is a highly aggressive skin cancer and currently resistant to systemic therapy. Melanomas may involve genetic, epigenetic and metabolic abnormalities. Evidence is emerging that epigenetic changes might play a significant role in tumor cell plasticity and metastatic phenotype of melanoma cells. PRINCIPAL FINDINGS: In this study, we developed a systematic approach to identify genes implicated in melanoma progression. To do this, we used the Affymetrix GeneChip Arrays to screen 34,000 mouse transcripts in melan-a melanocytes, 4C pre-malignant melanocytes, 4C11- non-metastatic and 4C11+ metastatic melanoma cell lines. The genome-wide association studies revealed pathways commonly over-represented in the transition from immortalized to pre-malignant stage, and under-represented in the transition from non-metastatic to metastatic stage. Additionally, the treatment of cells with 10 µM 5-aza-2'-deoxycytidine (5AzaCdR) for 48 hours allowed us to identify genes differentially re-expressed at specific stages of melan-a malignant transformation. Treatment of human primary melanocytes with the demethylating agent 5AzaCdR in combination to the histone deacetylase inhibitor Trichostatin A (TSA) revealed changes on melanocyte morphology and gene expression which could be an indicator of epigenetic flexibility in normal melanocytes. Moreover, changes on gene expression recognized by affecting the melanocyte biology (NDRG2 and VDR), phenotype of metastatic melanoma cells (HSPB1 and SERPINE1) and response to cancer therapy (CTCF, NSD1 and SRC) were found when Mel-2 and/or Mel-3-derived patient metastases were exposed to 5AzaCdR plus TSA treatment. Hierarchical clustering and network analyses in a panel of five patient-derived metastatic melanoma cells showed gene interactions that have never been described in melanomas. SIGNIFICANCE: Despite the heterogeneity observed in melanomas, this study demonstrates the utility of our murine melanoma progression model to identify molecular markers commonly perturbed in metastasis. Additionally, the novel gene expression signature identified here may be useful in the future into a model more closely related to translational research.

Ensino baseado na Web: desenvolvimento, implementaçäo e avaliaçäo de um curso de graduaçäo em medicina / Web-based learning: development, implementation and assessment of an undergraduate medical course

Objetivo: 0 objetivo do presente estudo foi desenvolver, implementar e avaliar um curso baseado na tecnologia Web em cirurgia experimental para alunos do curso de graduaçao em Medicina da Universidade Federal do Estado de Sao Paulo (UNIFESPEPM). 0 curso tradicional em cirurgia experimental, baseado em palestras formais e atividades laboratoriais, é oferecido na grade curricular do terceiro ano médico desta Universidade. A parte teórica do curso foi desenvolvida e implementada na Web e oferecida aos alunos no segundo semestre do ano de 2001. A efetividade do curso, o ganho em conhecimento obtido pelos alunos e suas atitudes em relaçao a este novo ambiente de aprendizado foram avaliados. Os processos de desenvolvimento e implementaçao deste novo formato de curso também foram descritos e discutidos. Métodos: A populaçao do estudo foi de 66 estudantes do terceiro ano médico desta Universidade que, durante cinco semanas, participaram do curso e o avaliaram. 0 desenho de pesquisa usado foi: Um Grupo Pré e Pós-teste Sem Grupo Controle. Seis questionários de avaliaçao baseados em uma escala de Likert de cinco pontos foram apresentados aos alunos no web site do curso. A efetividade do curso e o ganho dos alunos em conhecimento sobre cirurgia experimental foram determinados pela análise dos seguintes dados: os resultados das avaliaçoes, as notas dos testes pré e pós-curso e as variáveis do rastreamento dos alunos. Resultados: Os resultados demonstraram níveis altos de funcionalidade do curso e de efetividade do seu material. 0 grau de aceitaçao entre os estudantes também foi alto. 0 ganho em conhecimento foi estatisticamente significativo após o curso (p < 0,001). A correlaçao entre o ganho e o desempenho dos alunos na atividade interativa (notas do vídeo quiz), embora positiva e significativa, foi muito fraca. Este estudo apresentou evidências de que os vídeos interativos sao úteis para os cursos baseados na Web, especialmente por influenciarem no ganho em conhecimento. Nao houve correlaçao entre o ganho e o tempo que os alunos gastaram navegando nas páginas do curso. Conclusoes: o Curso Teórico de Técnica Operatória e Cirurgia Experimental para a graduaçao médica na Web foi desenvolvido e implementado com êxito. A avaliaçao realizada mostrou que o curso é efetivo e aceito entre os alunos. 0 material didático on-line de alta qualidade, os graus elevados de funcionalidade e de aceitaçao do curso...(au)