Primary colorectal diffuse large B-cell lymphoma: A report of eighteen cases in a tertiary care center.

Primary colorectal diffuse large B-cell lymphoma (DLBCL) is very rare colon malignancy. It is important to know the main demographic and clinical characteristics of these patients. We conducted a retrospective analysis of 18 patients diagnosed with primary colorectal DLBCL during a 17-year period at the National Cancer Institute of Brazil (INCA) between 2000 and 2018. Demographic characteristics, tumor localization, HIV status, lactate dehydrogenase (LDH) levels, treatment modality and follow-up status were obtained from medical records. Survival was estimated from the date of diagnosis until death. There were 11 male and seven female patients in our cohort, the median age at diagnosis was 59.5 years and four patients were HIV positive. Tumor was mainly localized in the right colon. Patients were treated with chemotherapy (CT) and/or surgical resection. Eleven patients died during a median follow-up of 59 months and the median survival time was 10 months. Six or more cycles of CT (HR=0.19; CI 95% 0.054-0.660, p = 0.009), LDH levels below 350 U/L (HR=0.229; CI 95% 0.060-0.876, p = 0.031) and surgical resection (HR=0.23; CI 95% 0.065-0.828, p = 0.030) were associated with reduced risk of death in univariate analysis. Patient's age and DLBCL right colon localization should be considered at diagnosis to distinguish between DLBCL and other diseases for differential diagnosis. Six cycles of CT, LDH levels below 350 U/L and surgical resection were associated with better survival. Our results are consistent with previous publications and address the importance of correct colorectal DLBCL diagnosis and treatment.

Primary diffuse large B-cell lymphoma of the head and neck in a Brazilian single-center study.

OBJECTIVES: Lymphomas represent around 10% of head and neck neoplasms, among which the diffuse large B-cell lymphoma (DLBCL) is the most common histologic subtype. In the present study, we characterized demographic parameters, anatomical sites, and survival rates of patients in a Brazilian cancer center. MATERIALS AND METHODS: Single-center retrospective epidemiological study of 243 head and neck DLBCL patients. Demographic characteristics, tumor localization, HIV status, lactate dehydrogenase (LDH) activity, and treatment modality were obtained from electronic medical records. RESULTS: The most common primary head and neck tumor location in patients with DLBCL was Waldeyer's ring. Interestingly, age above 80 years, male gender, high LDH levels, and HIV positivity were significantly associated with shorter overall survival (OS) rates and increased risk of death. We further demonstrated that treatment had a protective effect, improving OS, and reducing risk of death. Notably, we found no benefit of combination of chemotherapy and radiotherapy versus isolated treatment modalities. CONCLUSION: The study showed that primary head and neck DLBCL is more incident in middle age and elderly patients with a small male patients' majority in a Brazilian population. Moreover, we observed a 3-year OS rate of almost 60% and multivariate analysis showed that treatment was the only protective factor.

Crosstalk between biological and chemical diversity with cytotoxic and cytostatic effects of Aphanothece halophytica in vitro.

Different solvent extracts from Aphanothece halophytica (A. halophytica) were evaluated for their cytotoxic effects against four human cancer cell lines. The samples demonstrated different percentages of cyanobacteria species populations. The samples containing 100% A. halophytica and 90% A. halophytica showed a significant cytotoxic effect in human breast cancer cells MDA231. The cytostatic effect was demonstrated in MDA231 and human glioblastoma T98G cells regardless of the treatment, resulting in a significant cell cycle arrest in the S phase. The chemical profiles of the extracts were proven to be diverse in qualitative and quantitative compositions. This variability was dependent on the A. halophytica´s abundance in each extract. The 100% A. halophytica extract induced cytotoxic and cytostatic effects in breast cancer cells, and those could be associated with the predominance of fatty acids, hydrocarbons and phthalates, indicating that A. halophytica is an interesting source of novel compound with anticancer effect.

O que mudou entre os períodos de pico e de platô durante a primeira onda do SARS-CoV-2? Estudo multicêntrico português em unidades de cuidados intensivos / What changed between the peak and plateau periods of the first COVID-19 pandemic wave? A multicentric Portuguese cohort study in intensive care

RESUMO Objetivo: Analisar e comparar as características de pacientes críticos com a COVID-19, a abordagem clínica e os resultados entre os períodos de pico e de platô na primeira onda pandêmica em Portugal. Métodos: Este foi um estudo de coorte multicêntrico ambispectivo, que incluiu pacientes consecutivos com a forma grave da COVID-19 entre março e agosto de 2020 de 16 unidades de terapia intensiva portuguesas. Definiram-se as semanas 10 - 16 e 17 - 34 como os períodos de pico e platô. Resultados: Incluíram-se 541 pacientes adultos com mediana de idade de 65 [57 - 74] anos, a maioria do sexo masculino (71,2%). Não houve diferenças significativas na mediana de idade (p = 0,3), no Simplified Acute Physiology Score II (40 versus 39; p = 0,8), na pressão parcial de oxigênio/fração inspirada de oxigênio (139 versus 136; p = 0,6), na terapia com antibióticos na admissão (57% versus 64%; p = 0,2) ou na mortalidade aos 28 dias (24,4% versus 22,8%; p = 0,7) entre o período de pico e platô. Durante o período de pico, os pacientes tiveram menos comorbidades (1 [0 - 3] versus 2 [0 - 5]; p = 0,002); fizeram mais uso de vasopressores (47% versus 36%; p < 0,001) e ventilação mecânica invasiva na admissão (58,1% versus 49,2%; p < 0,001), e tiveram mais prescrição de hidroxicloroquina (59% versus 10%; p < 0,001), lopinavir/ritonavir (41% versus 10%; p < 0,001) e posição prona (45% versus 36%; p = 0,04). Entretanto, durante o platô, observou-se maior uso de cânulas nasais de alto fluxo (5% versus 16%; p < 0,001) na admissão, remdesivir (0,3% versus 15%; p < 0,001) e corticosteroides (29% versus 52%; p < 0,001), além de menor tempo de internação na unidade de terapia intensiva (12 versus 8 dias; p < 0,001). Conclusão: Houve mudanças significativas nas comorbidades dos pacientes, nos tratamentos da unidade de terapia intensiva e no tempo de internação entre os períodos de pico e platô na primeira onda da COVID-19.

ABSTRACT Objective: To analyze and compare COVID-19 patient characteristics, clinical management and outcomes between the peak and plateau periods of the first pandemic wave in Portugal. Methods: This was a multicentric ambispective cohort study including consecutive severe COVID-19 patients between March and August 2020 from 16 Portuguese intensive care units. The peak and plateau periods, respectively, weeks 10 - 16 and 17 - 34, were defined. Results: Five hundred forty-one adult patients with a median age of 65 [57 - 74] years, mostly male (71.2%), were included. There were no significant differences in median age (p = 0.3), Simplified Acute Physiology Score II (40 versus 39; p = 0.8), partial arterial oxygen pressure/fraction of inspired oxygen ratio (139 versus 136; p = 0.6), antibiotic therapy (57% versus 64%; p = 0.2) at admission, or 28-day mortality (24.4% versus 22.8%; p = 0.7) between the peak and plateau periods. During the peak period, patients had fewer comorbidities (1 [0 - 3] versus 2 [0 - 5]; p = 0.002) and presented a higher use of vasopressors (47% versus 36%; p < 0.001) and invasive mechanical ventilation (58.1 versus 49.2%; p < 0.001) at admission, prone positioning (45% versus 36%; p = 0.04), and hydroxychloroquine (59% versus 10%; p < 0.001) and lopinavir/ritonavir (41% versus 10%; p < 0.001) prescriptions. However, a greater use of high-flow nasal cannulas (5% versus 16%, p < 0.001) on admission, remdesivir (0.3% versus 15%; p < 0.001) and corticosteroid (29% versus 52%, p < 0.001) therapy, and a shorter ICU length of stay (12 days versus 8, p < 0.001) were observed during the plateau. Conclusion: There were significant changes in patient comorbidities, intensive care unit therapies and length of stay between the peak and plateau periods of the first COVID-19 wave.

Bioremediation of hydrocarbon contaminated soil using local organic materials and earthworms.

Bioremediation technologies have demonstrated significant success on biological quality recovery of hydrocarbon contaminated soils, employing techniques among which composting and vermiremediation stand out. The aim of this study was to evaluate the efficiency of these processes to remediate diesel-contaminated soil, employing local organic materials and earthworms. During the initial composting stage (75 days), the substrate was made up using contaminated soil, lombricompost, rice hulls and wheat stubbles (60:20:15:5% w/w). Diesel concentration in the contaminated substrate was about 5 g kg-1, equivalent to a Total Petroleum Hidrocarbons (TPH) experimental concentration of 3425 ± 50 mg kg-1. During the later vermiremediation stage (60 days), the earthworm species Eisenia fetida and Amynthas morrisi were evaluated for their hydrocarbon degradation capacity. Physicochemical and biological assays were measured at different times of each stage and ecotoxicity assays were performed at the end of the experiments. TPH concentration reduced 10.91% after composting and from 45.2 to 60.81% in the different treatments after vermiremediation. Compared with TPH degradation in the treatment without earthworms (16.05%), results indicate that earthworms, along with indigenous microorganisms, accelerate the remediation process. Vermiremediation treatments did not present phytotoxicity and reflected high substrate maturity values (>80% Germination Index) although toxic effects were observed due to E. fetida and A morrisi exposure to diesel. Vermiremediation was an efficient technology for the recovery of substrate biological quality after diesel contamination in a short period. The addition of organic materials and suitable food sources aided earthworm subsistence, promoted the decontamination process and improved the substrate quality for future productive applications.

Effects of long-term exposure to MST-312 on lung cancer cells tumorigenesis: Role of SHH/GLI-1 axis.

Replicative immortality is a key feature of cancer cells and it is maintained by the expression of telomerase, a promising target of novel therapies. Long-term telomerase inhibition can induce resistance, but the mechanisms underlying this process remain unclear. The Sonic hedgehog pathway (SHH) is an embryogenic pathway involved in tumorigenesis and modulates the transcription of telomerase. We evaluated the effects of long-term treatment of the telomerase inhibitor MST-312 in morphology, proliferation, resistance, and in the SHH pathway molecules expression levels in lung cancer cells. Cells treated for 12 weeks with MST-312 showed changes in morphology, such as spindle-shaped cells, and a shift in the distribution of F-ACTIN from cortical to diffuse. Treatment also significantly reduced cells' efficiency to form spheroids and their clonogenic potential, independently of the cell cycle and telomeric DNA content. Moreover, GLI-1 expression levels were significantly reduced after 12 weeks of MST-312 treatment, indicating a possible inhibition of this signaling axis in the SHH pathway, without hindering NANOG and OCT4 expression. Here, we described a novel implication of long-term treatment with MST-312 functionally and molecularly, shedding new light on the molecular mechanisms of this drug in vitro.

What changed between the peak and plateau periods of the first COVID-19 pandemic wave? A multicentric Portuguese cohort study in intensive care. / O que mudou entre os períodos de pico e de platô durante a primeira onda do SARS-CoV-2? Estudo multicêntrico português em unidades de cuidados intensivos.

OBJECTIVE: To analyze and compare COVID-19 patient characteristics, clinical management and outcomes between the peak and plateau periods of the first pandemic wave in Portugal. METHODS: This was a multicentric ambispective cohort study including consecutive severe COVID-19 patients between March and August 2020 from 16 Portuguese intensive care units. The peak and plateau periods, respectively, weeks 10 - 16 and 17 - 34, were defined. RESULTS: Five hundred forty-one adult patients with a median age of 65 [57 - 74] years, mostly male (71.2%), were included. There were no significant differences in median age (p = 0.3), Simplified Acute Physiology Score II (40 versus 39; p = 0.8), partial arterial oxygen pressure/fraction of inspired oxygen ratio (139 versus 136; p = 0.6), antibiotic therapy (57% versus 64%; p = 0.2) at admission, or 28-day mortality (24.4% versus 22.8%; p = 0.7) between the peak and plateau periods. During the peak period, patients had fewer comorbidities (1 [0 - 3] versus 2 [0 - 5]; p = 0.002) and presented a higher use of vasopressors (47% versus 36%; p < 0.001) and invasive mechanical ventilation (58.1 versus 49.2%; p < 0.001) at admission, prone positioning (45% versus 36%; p = 0.04), and hydroxychloroquine (59% versus 10%; p < 0.001) and lopinavir/ritonavir (41% versus 10%; p < 0.001) prescriptions. However, a greater use of high-flow nasal cannulas (5% versus 16%, p < 0.001) on admission, remdesivir (0.3% versus 15%; p < 0.001) and corticosteroid (29% versus 52%, p < 0.001) therapy, and a shorter ICU length of stay (12 days versus 8, p < 0.001) were observed during the plateau. CONCLUSION: There were significant changes in patient comorbidities, intensive care unit therapies and length of stay between the peak and plateau periods of the first COVID-19 wave.

OBJETIVO: Analisar e comparar as características de pacientes críticos com a COVID-19, a abordagem clínica e os resultados entre os períodos de pico e de platô na primeira onda pandêmica em Portugal. MÉTODOS: Este foi um estudo de coorte multicêntrico ambispectivo, que incluiu pacientes consecutivos com a forma grave da COVID-19 entre março e agosto de 2020 de 16 unidades de terapia intensiva portuguesas. Definiram-se as semanas 10 - 16 e 17 - 34 como os períodos de pico e platô. RESULTADOS: Incluíram-se 541 pacientes adultos com mediana de idade de 65 [57 - 74] anos, a maioria do sexo masculino (71,2%). Não houve diferenças significativas na mediana de idade (p = 0,3), no Simplified Acute Physiology Score II (40 versus 39; p = 0,8), na pressão parcial de oxigênio/fração inspirada de oxigênio (139 versus 136; p = 0,6), na terapia com antibióticos na admissão (57% versus 64%; p = 0,2) ou na mortalidade aos 28 dias (24,4% versus 22,8%; p = 0,7) entre o período de pico e platô. Durante o período de pico, os pacientes tiveram menos comorbidades (1 [0 - 3] versus 2 [0 - 5]; p = 0,002); fizeram mais uso de vasopressores (47% versus 36%; p < 0,001) e ventilação mecânica invasiva na admissão (58,1% versus 49,2%; p < 0,001), e tiveram mais prescrição de hidroxicloroquina (59% versus 10%; p < 0,001), lopinavir/ritonavir (41% versus 10%; p < 0,001) e posição prona (45% versus 36%; p = 0,04). Entretanto, durante o platô, observou-se maior uso de cânulas nasais de alto fluxo (5% versus 16%; p < 0,001) na admissão, remdesivir (0,3% versus 15%; p < 0,001) e corticosteroides (29% versus 52%; p < 0,001), além de menor tempo de internação na unidade de terapia intensiva (12 versus 8 dias; p < 0,001). CONCLUSÃO: Houve mudanças significativas nas comorbidades dos pacientes, nos tratamentos da unidade de terapia intensiva e no tempo de internação entre os períodos de pico e platô na primeira onda da COVID-19.

Amamentação em menores de dois anos em uma cidade da Região Amazônica / Lactancia en menores de dos años en una ciudad de la Región Amazónica / Breastfeeding in infants under two years old in a city in the Amazon Region

Resumo Objetivo Analisar a duração do aleitamento materno e os fatores associados ao desmame total de crianças de seis a 23 meses e 29 dias de idade residentes no município de Cruzeiro do Sul, na Amazônia Ocidental Brasileira. Métodos Estudo transversal, realizado durante a Campanha Nacional de Multivacinação em 2016 e Contra a Influenza em 2017. A amostra foi calculada por conglomerados. A coleta de dados foi efetuada com as mães ou os responsáveis de 679 crianças que compareceram às campanhas de vacinação e responderam a um questionário. Utilizou-se a análise de sobrevivência de Kaplan-Meier e, para os fatores associados ao desmame total, a regressão de Cox. Para todos os testes estatísticos, foi considerado um nível de significância de 5%. Resultados O aleitamento materno foi praticado por 65,3% das crianças, cuja média de idade foi de 13,7 meses (DP± 4,9 meses). O tempo médio de desmame total foi de 16,7 meses (IC95%: 16,06 - 17,36) e a mediana de 22 meses, sendo a probabilidade de tempo de aleitamento materno até dois anos em 49,7%. Os fatores associados ao desmame total foram o tempo da experiência anterior em amamentação menor que seis meses, não praticar o aleitamento materno na primeira hora de vida, uso de chupeta e mamadeira. Conclusão A duração do aleitamento materno foi abaixo do recomendado. Os fatores associados ao desmame total de crianças entre 6 e 23 meses estão relacionados à experiência materna prévia, ao início precoce da prática de amamentação e ao uso de bicos artificiais.

Resumen Objetivo Analizar la duración de la lactancia materna y los factores asociados al destete total de niños de seis meses a 23 meses y 29 días de edad que viven en el municipio de Cruzeiro do Sul, en la Amazonía Occidental Brasileña. Métodos Estudio transversal, realizado durante la Campaña Nacional de Multivacunación en 2016 y Contra la Influenza en el 2017. La muestra fue calculada por conglomerados. La recopilación de datos se realizó con las madres o los responsables de 679 niños que asistieron a las campañas de vacunación y respondieron un cuestionario. Se utilizó el análisis de supervivencia de Kaplan-Meier y, para los factores asociados al destete total, la regresión de Cox. Para todas las pruebas estadísticas se consideró un nivel de significación del 5 %. Resultados La lactancia materna se practicó en el 65,3 % de los niños, cuyo promedio de edad fue de 13,7 meses (DP± 4,9 meses). El tiempo promedio de destete total fue de 16,7 meses (IC95 %: 16,06 - 17,36) y la mediana de 22 meses, con una probabilidad de tiempo de lactancia materna hasta los dos años del 49,7 %. Los factores asociados al destete total fueron el tiempo de una experiencia anterior de lactancia inferior a seis meses, no practicar la lactancia materna en la primera hora de vida, uso de chupete y de mamadera. Conclusión La duración de la lactancia materna estuvo por debajo de lo recomendado. Los factores asociados al destete total de niños entre 6 y 23 meses están relacionados a experiencias maternas previas, al inicio precoz de la práctica de la lactancia y al uso de tetinas artificiales.

Abstract Objective To analyze the duration of breastfeeding and the factors associated with total weaning of infants aged six to 23 months and 29 days living in the municipality of Cruzeiro do Sul, in the Brazilian Western Amazon. Methods A cross-sectional study conducted during the National Multi-Vaccination Campaign in 2016 and Against Influenza in 2017. The sample was estimated by clusters. The data was collected with the mothers or guardians of 679 infants who attended vaccination campaigns and answered a questionnaire. Kaplan-Meier survival analysis was used, and Cox regression was used for the factors associated with total weaning. A 5% significance level was considered for all statistical tests. Results Breastfeeding was practiced by 65.3% of the infants, whose mean age was 13.7 months (SD± 4.9 months). The mean total weaning time was 16.7 months (95% CI: 16.06 - 17.36) and the median of 22 months, which is the probability of duration of breastfeeding up to two years old in 49.7%. The factors associated with total weaning were previous breastfeeding experience for less than six months, not breastfeeding in the first hour of life, pacifier use and bottle-feeding. Conclusion The duration of breastfeeding was below the recommended. The factors associated with total weaning of infants between six and 23 months are related to previous maternal experience, early initiation of breastfeeding and the use of artificial nipples.

Low rate of relapse after twelve-dose multidrug therapy for hansen's disease: A 20-year cohort study in a brazilian reference center.

The World Health Organization has raised concerns about the increasing number of Hansen disease (HD) relapses worldwide, especially in Brazil, India, and Indonesia that report the highest number of recurrent cases. Relapses are an indicator of MDT effectiveness and can reflect Mycobacterium leprae persistence or re-infection. Relapse is also a potential marker for the development or progression of disability. In this research, we studied a large cohort of persons affected by HD treated with full fixed-dose multibacillary (MB) multidrug therapy (MDT) followed for up to 20 years and observed that relapses are a rare event. We estimated the incidence density of relapse in a cohort of patients classified to receive MB regime (bacillary index (BI) > 0), diagnosed between September 1997 and June 2017, and treated with twelve-dose MB-MDT at a HD reference center in Rio de Janeiro, Brazil. We obtained the data from the data management system of the clinic routine service. We linked the selected cases to the dataset of relapses of the national HD data to confirm possible relapse cases diagnosed elsewhere. We diagnosed ten cases of relapse in a cohort of 713 patients followed-up for a mean of 12.1 years. This resulted in an incidence rate of 1.16 relapse cases per 1000 person-year (95% CI = 0.5915-2.076). The accumulated risk was 0.025 in 20 years. The very low risk observed in this cohort of twelve-dose-treated MB patients reinforces the success of the current MDT scheme.

The pterocarpanquinone LQB­118 compound induces apoptosis of cytarabine­resistant acute myeloid leukemia cells.

Acute myeloid leukemia (AML) is a complex hematological disorder characterized by blockage of differentiation and high proliferation rates of myeloid progenitors. Anthracycline and cytarabine­based therapy has remained the standard treatment for AML over the last four decades. Although this treatment strategy has increased survival rates, patients often develop resistance to these drugs. Despite efforts to understand the mechanisms underlying cytarabine resistance, there have been few advances in the field. The present study developed an in vitro AML cell line model resistant to cytarabine (HL­60R), and identified chromosomal aberrations by karyotype evaluation and potential molecular mechanisms underlying chemoresistance. Cytarabine decreased cell viability, as determined by MTT assay, and induced cell death and cell cycle arrest in the parental HL­60 cell line, as revealed by Annexin V/propidium iodide (PI) staining and PI DNA incorporation, respectively, whereas no change was observed in the HL­60R cell line. In addition, the HL­60R cell line exhibited a higher tumorigenic capacity in vivo compared with the parental cell line. Notably, no reduction in tumor volume was detected in mice treated with cytarabine and inoculated with HL­60R cells. In addition, western blotting revealed that the protein expression levels of Bcl­2, X­linked inhibitor of apoptosis protein (XIAP) and c­Myc were upregulated in HL­60R cells compared with those in HL­60 cells, along with predominant nuclear localization of the p50 and p65 subunits of NF­κB in HL­60R cells. Furthermore, the antitumor effect of LQB­118 pterocarpanquinone was investigated; this compound induced apoptosis, a reduction in cell viability and a decrease in XIAP expression in cytarabine­resistant cells. Taken together, these data indicated that acquired cytarabine resistance in AML was a multifactorial process, involving chromosomal aberrations, and differential expression of apoptosis and cell proliferation signaling pathways. Furthermore, LQB­118 could be a potential alternative therapeutic approach to treat cytarabine­resistant leukemia cells.

Update on drug transporter proteins in acute myeloid leukemia: Pathological implication and clinical setting.

Acute myeloid leukemia (AML) is one of the most common hematological neoplasia causing death worldwide. The long-term overall survival is unsatisfactory due to many factors including older age, genetic heterogeneity and molecular characteristics comprising additional mutations, and resistance to chemotherapeutic drugs. The expression of ABCB1/P-glycoprotein, ABCC1/MRP1, ABCG2/BCRP and LRP transporter proteins is considered the major reason for multidrug resistance (MDR) in AML, however conflicting data have been reported. Here, we review the main issues about drug transporter proteins in AML clinical scenario, and highlight the clinicopathological significance of MDR phenotype associated with ABCB1 polymorphisms and FLT3 mutation.

Osteopontin­c isoform inhibition modulates ovarian cancer cell cisplatin resistance, viability and plasticity.

Osteopontin (OPN) is upregulated in several types of tumor and has been associated with chemoresistance. However, the contribution of OPN splicing isoforms (OPN­SIs) to chemoresistance requires further investigation. The present study aimed to evaluate the expression patterns of each tested OPN­SI in cisplatin (CDDP)­resistant ovarian carcinoma cell lines, focusing on the role of the OPN­c isoform (OPNc) in drug resistance. ACRP ovarian cancer cells resistant to CDDP, as well as their parental cell line A2780, were used. Analyses of the transcriptional expression of OPN­SIs, epithelial­mesenchymal transition (EMT) markers and EMT­related cytokines were performed using reverse transcription­quantitative PCR. OPNc was silenced in ACRP cells using anti­OPNc DNA oligomers and stably overexpressed by transfecting A2780 cells with a mammalian expression vector containing the full length OPNc cDNA. Functional assays were performed to determine cell proliferation, viability and colony formation. The results demonstrated that among the three tested OPN­SIs, OPNc was the most upregulated transcript in the ACRP cells compared with the parental A2780 cells. In addition, the expression levels of P­glycoprotein multidrug transporter were upregulated in CDDP­resistant ACRP cells compared with those in A2780 cells. OPNc knockdown sensitized ACRP cells to CDDP treatment and downregulated P­gp expression levels compared with those in the negative control group. Additionally, silencing of OPNc impaired cell proliferative and colony formation abilities, as well as reversed the expression levels of EMT markers and EMT­related cytokines compared with those in the negative control cells. Notably, although stable OPNc overexpression resulted in increased A2780 cell proliferation, it notably increased CDDP sensitivity compared with that in the cells transfected with a control vector. These results suggested that OPNc silencing may represent a putative approach to sensitize resistant ovarian cancer cells to chemotherapeutic agents.

Detection of TNF-α Protein in Extracellular Vesicles Derived from Tumor Cells by Western Blotting.

Detection of tumor necrosis factor-alpha (TNF-α) is usually performed in cell cultured medium or body fluids via measurement of its soluble extracellular form. However, depending on cellular condition, TNF-α might be transported through extracellular vesicles (EV) from donor cells to recipient cells. EV are small membrane-delimited structures (∼50 nm to 10 µm) that are spontaneously released from multiple cell types. In cancer, EV arise as important mediators in intercellular communication, and their molecular content may support tumor progression. This chapter describes methods to identify protein content in EV released from the tumor cell cultures. Through this protocol, we show first how to purify EV from in vitro cell culture by using differential centrifugation technique and then we demonstrate how to identify both membrane and soluble TNF-α forms in EV by Western blotting.

Agreement between nutritional status and perception of body image in indigenous Khisêdjê of the Xingu Indigenous Park. / Concordância entre estado nutricional e percepção da imagem corporal em indígenas khisêdjê do Parque Indígena do Xingu.

OBJECTIVE: To determine the agreement between body self-image (based on the Stunkard figure rating scale) and nutritional status and to evaluate body satisfaction among the Khisêdjê indigenous people of Parque Indígena do Xingu (Xingu Indigenous Park). METHODS: A cross-sectional study involving 131 natives aged 20 and older. Data on body image, body mass index and waist circumference were collected. Kappa statistics, χ2 (p < 0.05), crude and adjusted prevalence ratios and Student's t-test were used for data analysis. RESULTS: The prevalence of overweight and obesity was respectively 42 and 5.3%. The percentage of satisfaction with body profile was 61.8% with no difference between the sexes. There was good agreement between actual and ideal self-image (p < 0.001), but poor agreement between actual and ideal self-image with nutritional status for both sexes. A higher prevalence of body dissatisfaction due to overweight was detected in individuals with central obesity and overweight. CONCLUSION: The results suggest that body self-image evaluated by the Stunkard silhouette scale has little applicability as an indicator of nutritional status among the indigenous Khisêdjê of Xingu Indigenous Park.

Expression of nuclear XIAP associates with cell growth and drug resistance and confers poor prognosis in breast cancer.

Evasion from apoptosis is one of the hallmarks of cancer. X-linked inhibitor of apoptosis protein (XIAP) is known to modulate apoptosis by inhibiting caspases and ubiquitinating target proteins. XIAP is mainly found at the cytoplasm, but recent data link nuclear XIAP to poor prognosis in breast cancer. Here, we generated a mutant form of XIAP with a nuclear localization signal (XIAPNLS-C-term) and investigated the oncogenic mechanisms associated with nuclear XIAP in breast cancer. Our results show that cells overexpressing XIAPΔRING (RING deletion) and XIAPNLS-C-term exhibited XIAP nuclear localization more abundantly than XIAPwild-type. Remarkably, overexpression of XIAPNLS-C-term, but not XIAPΔRING, conferred resistance to doxorubicin and increased cellular proliferative capacity. Interestingly, Survivin and c-IAP1 expression were not associated with XIAP oncogenic effects. However, NFκB expression and ubiquitination of K63, but not K48 chains, were increased following XIAPNLS-C-term overexpression, pointing to nuclear signaling transduction. Consistently, multivariate analysis revealed nuclear, but not cytoplasmic XIAP, as an independent prognostic factor in hormone receptor-negative breast cancer patients. Altogether, our findings suggest that nuclear XIAP confers poor outcome and RING-associated breast cancer growth and chemoresistance.

The Effect of Nanosystems on ATP-Binding Cassette Transporters: Understanding the Influence of Nanosystems on Multidrug Resistance Protein-1 and P-glycoprotein.

The cancer multidrug resistance is involved in the failure of several treatments during cancer treatment. It is a phenomenon that has been receiving great attention in the last years due to the sheer amount of mechanisms discovered and involved in the process of resistance which hinders the effectiveness of many anti-cancer drugs. Among the mechanisms involved in the multidrug resistance, the participation of ATP-binding cassette (ABC) transporters is the main one. The ABC transporters are a group of plasma membrane and intracellular organelle proteins involved in the process of externalization of substrates from cells, which are expressed in cancer. They are involved in the clearance of intracellular metabolites as ions, hormones, lipids and other small molecules from the cell, affecting directly and indirectly drug absorption, distribution, metabolism and excretion. Other mechanisms responsible for resistance are the signaling pathways and the anti- and pro-apoptotic proteins involved in cell death by apoptosis. In this study we evaluated the influence of three nanosystem (Graphene Quantum Dots (GQDs), mesoporous silica (MSN) and poly-lactic nanoparticles (PLA)) in the main mechanism related to the cancer multidrug resistance such as the Multidrug Resistance Protein-1 and P-glycoprotein. We also evaluated this influence in a group of proteins involved in the apoptosis-related resistance including cIAP-1, XIAP, Bcl-2, BAK and Survivin proteins. Last, colonogenic and MTT (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assays have also been performed. The results showed, regardless of the concentration used, GQDs, MSN and PLA were not cytotoxic to MDA-MB-231 cells and showed no impairment in the colony formation capacity. In addition, it has been observed that P-gp membrane expression was not significantly altered by any of the three nanomaterials. The results suggest that GQDs nanoparticles would be suitable for the delivery of other multidrug resistance protein 1 (MRP1) substrate drugs that bind to the transporter at the same binding pocket, while MSN can strongly inhibit doxorubicin efflux by MRP1. On the other hand, PLA showed moderate inhibition of doxorubicin efflux by MRP1 suggesting that this nanomaterial can also be useful to treat MDR (Multidrug resistance) due to MRP1 overexpression.

Unraveling survivin expression in chronic myeloid leukemia: Molecular interactions and clinical implications.

Chronic myeloid leukemia (CML) is a myeloproliferative disorder characterized by the BCR-ABL oncoprotein, known to drive leukemogenesis by orchestrating multiple signaling pathways ultimately involved in cell survival. Despite successful response rates of CML patients to tyrosine kinase inhibitors (TKIs), resistance eventually arises due to BCR-ABL-dependent and independent mechanisms. Survivin is an inhibitor of apoptosis protein acting in the interface between apoptosis deregulation and cell cycle progression. In CML, high levels of survivin have been associated with late stages of disease and therapy resistance. In this review, we provide an overview of important aspects concerning survivin subcellular localization and expression pattern in CML patients and cell lines. Moreover, we highlight the relevance of molecular networks involving survivin for disease progression and treatment resistance. Finally, we discuss the mechanisms accounting for survivin overexpression, as well as novel therapeutic interventions that have been designed to counteract survivin-associated malignancy in CML.

LQB­118 compound inhibits migration and induces cell death in glioblastoma cells.

Glioblastoma (GBM) is the most frequent malignant brain tumor. It represents the most aggressive astrocytoma with an overall survival of 14 months. Despite improvements in surgery techniques, radio­ and chemotherapy, most patients present treatment resistance, recurrence and disease progression. Therefore, development of effective alternative therapies is essential to overcome treatment failure. The purpose of the study was to evaluate the antitumoral activity of the synthetic compound LQB­118, in vitro. Monolayer and three­dimensional (3D) cell culture systems of human­derived GBM cell lines were used to evaluate the effect of LQB­118 on cell viability, cell death and migration. LQB­118 reduced cell viability as determined by MTT and trypan blue exclusion assays and promoted apoptosis in monolayer cell lines with an intrinsic temozolomide (TMZ)­resistance profile. In 3D culture models, LQB­118 reduced cell viability as evaluated by APH assay and inhibited cell migration while the TMZ resistance profile was maintained. Moreover, LQB­118 reduced p38 and AKT expression and phosphorylation, whereas it reduced only the phosphorylated ERK1/2 form. LQB­118 reduced p38 and NRF2 expression, an axis that is associated with TMZ resistance, revealing a mechanism to overcome resistance. LQB­118 also demonstrated an additional effect when combined with ionizing radiation and cisplatin. In conclusion, the present data demonstrated that LQB­118 maintained its effectiveness in a 3D cell conformation, which shares more similarities with the tumor mass. LQB­118 is a promising agent for GBM treatment as monotherapy and associated with radiotherapy or cisplatin. Its effect is associated with inhibition of GBM­related survival signaling pathways.

Concordância entre estado nutricional e percepção da imagem corporal em indígenas khisêdjê do Parque Indígena do Xingu / Agreement between nutritional status and perception of body image in indigenous Khisêdjê of the Xingu Indigenous Park

RESUMO: Objetivos: Verificar a concordância entre autoimagem corporal (escala de silhuetas de Stunkard et al.) e estado nutricional e avaliar a satisfação corporal em indígenas khisêdjê do Parque Indígena do Xingu. Métodos: Estudo transversal que incluiu 131 indígenas khisêdjê, com 20 anos ou mais. Coletaram-se dados sobre imagem corporal, índice de massa corporal e perímetro da cintura. Foram utilizados a estatística kappa, o teste χ2 (p < 0,05), as razões de prevalências brutas e ajustadas e o teste t de Student. Resultados: As prevalências de sobrepeso e obesidade foram, respectivamente, 42 e 5,3%. A porcentagem de satisfação com o perfil corporal foi de 61,8%, sem diferença entre os sexos. Houve boa concordância entre autoimagem real e autoimagem ideal (p < 0,001), porém baixa concordância entre a autoimagem real e ideal e o estado nutricional para ambos os sexos. Maior prevalência de insatisfação corporal por excesso de peso foi detectada entre indivíduos com obesidade central e excesso de peso. Conclusão: Os resultados sugerem que a autoimagem corporal avaliada por meio da escala de silhuetas de Stunkard et al. tem pouca aplicabilidade como indicador do estado nutricional de indígenas khisêdjê do Parque Indígena do Xingu.

ABSTRACT: Objective: To determine the agreement between body self-image (based on the Stunkard figure rating scale) and nutritional status and to evaluate body satisfaction among the Khisêdjê indigenous people of Parque Indígena do Xingu (Xingu Indigenous Park). Methods: A cross-sectional study involving 131 natives aged 20 and older. Data on body image, body mass index and waist circumference were collected. Kappa statistics, χ2 (p < 0.05), crude and adjusted prevalence ratios and Student's t-test were used for data analysis. Results: The prevalence of overweight and obesity was respectively 42 and 5.3%. The percentage of satisfaction with body profile was 61.8% with no difference between the sexes. There was good agreement between actual and ideal self-image (p < 0.001), but poor agreement between actual and ideal self-image with nutritional status for both sexes. A higher prevalence of body dissatisfaction due to overweight was detected in individuals with central obesity and overweight. Conclusion: The results suggest that body self-image evaluated by the Stunkard silhouette scale has little applicability as an indicator of nutritional status among the indigenous Khisêdjê of Xingu Indigenous Park.

The LQB-223 Compound Modulates Antiapoptotic Proteins and Impairs Breast Cancer Cell Growth and Migration.

Drug resistance represents a major issue in treating breast cancer, despite the identification of novel therapeutic strategies, biomarkers, and subgroups. We have previously identified the LQB-223, 11a-N-Tosyl-5-deoxi-pterocarpan, as a promising compound in sensitizing doxorubicin-resistant breast cancer cells, with little toxicity to non-neoplastic cells. Here, we investigated the mechanisms underlying LQB-223 antitumor effects in 2D and 3D models of breast cancer. MCF-7 and MDA-MB-231 cells had migration and motility profile assessed by wound-healing and phagokinetic track motility assays, respectively. Cytotoxicity in 3D conformation was evaluated by measuring spheroid size and performing acid phosphatase and gelatin migration assays. Protein expression was analyzed by immunoblotting. Our results show that LQB-223, but not doxorubicin treatment, suppressed the migratory and motility capacity of breast cancer cells. In 3D conformation, LQB-223 remarkably decreased cell viability, as well as reduced 3D culture size and migration. Mechanistically, LQB-223-mediated anticancer effects involved decreased proteins levels of XIAP, c-IAP1, and Mcl-1 chemoresistance-related proteins, but not survivin. Survivin knockdown partially potentiated LQB-223-induced cytotoxicity. Additionally, cell treatment with LQB-223 resulted in changes in the mRNA levels of epithelial-mesenchymal transition markers, suggesting that it might modulate cell plasticity. Our data demonstrate that LQB-223 impairs 3D culture growth and migration in 2D and 3D models of breast cancer exhibiting different phenotypes.