María Aparecida Santos e Campos

DETECTION OF BREAST CANCER CELLS IN THE PERIPHERAL BLOOD IS POSITIVELY CORRELATED WITH ESTROGEN-RECEPTOR STATUS AND PREDICTS FOR POOR PROGNOSIS

We investigated whether detection of cytokeratin‐positive (CK+) cells in the peripheral blood (PB) of breast cancer patients before chemotherapy could be a prognostic factor. Blood from a total of 92 breast cancer patients was evaluated for the presence of CK+ cells. Blood samples were collected before chemotherapy. Patients entered in the study included: neoadjuvant (n = 25), adjuvant (n = 42) and metastatic (n = 25). Blood samples (10 ml) were centrifuged using a double density‐gradient to recovering the mononuclear cell (MNC) and granulocyte cell (GC) fractions. Subsequently, positive immunomagnetic cell separation was carried out to isolating CK+ cells. The enriched cell fraction was cytocentrifuged and then immunocytochemically labeled using an anti‐cytokeratin antibody. Our results indicated that breast tumor cells sediment with both MNC and GC fractions. We therefore recommend examination of both fractions in all enrichment protocols. CK+ cells in PB were identified in 57 of 92 (62%) patients when MNC and GC fractions were assessed (range = 1–61 cells, median = 8). No CK+ cells were detected in blood samples of 16 healthy donors. There were significant differences in the presence of CK+ cells according to estrogen receptor expression (p = 0.049), and lymph node status (p = 0.033), but not to the age, menopausal status, type of patient (neoadjuvant, adjuvant or metastatic), TNM stage, histological type, progesterone receptor expression, c‐erbB2 expression, p53 expression or Ki67 expression. Regarding the relationship between tumor size (T) and the presence of CK+ cells, a borderline significant trend was observed (p = 0.07). The median follow‐up of the patients was 21 months and statistical analysis (Kaplan‐Meier analysis) showed that using the method we present, the detection of CK+ cells in PB before starting the chemotherapy in breast cancer patients was significantly correlated with both progression‐free survival (p = 0.058) and overall survival (p = 0.003). In conclusion, the present study suggests that detection of CK+ cells in PB before chemotherapy might identify breast cancer patients with poor prognosis.

Squalene protects against oxidative DNA damage in MCF10A human mammary epithelial cells but not in MCF7 and MDA-MB-231 human breast cancer cells

Until now, very little has been known about the antioxidant capacity of squalene and its effect on human breast tumourigenesis. In the present work, we investigated squalenes scavenging properties and its effect on cell proliferation, cell cycle profile, apoptosis, reactive oxygen species (ROS) level and oxidative DNA damage, using human breast cell lines. Our results showed that squalene neither possesses scavenging activity nor significantly alters cell proliferation rates, the cell cycle profile or cell apoptosis in human mammary epithelial cells (MCF10A), minimally invasive (MDA-MB-231) breast cancer cells, and highly invasive (MCF7) breast cancer cells. However, we found that squalene did exert the following effects on MCF10A epithelial cells in a dose-dependent manner: (a) it decreased intracellular ROS level, (b) it prevented H(2)O(2)-induced oxidative injury, and (c) it protected against oxidative DNA damage. Interestingly, squalene did not exert these effects on MCF7 and MDA-MB-231 cancer cells. Therefore, our data suggest that squalene, found in high amounts in virgin olive oils, could be partially responsible for the lower incidence of breast cancer in populations that consume the Mediterranean diet due to its protective activity against oxidative DNA damage in normal mammary cells.

Hydroxytyrosol Protects against Oxidative DNA Damage in Human Breast Cells

Over recent years, several studies have related olive oil ingestion to a low incidence of several diseases, including breast cancer. Hydroxytyrosol and tyrosol are two of the major phenols present in virgin olive oils. Despite the fact that they have been linked to cancer prevention, there is no evidence that clarifies their effect in human breast tumor and non-tumor cells. In the present work, we present hydroxytyrosol and tyrosol’s effects in human breast cell lines. Our results show that hydroxytyrosol acts as a more efficient free radical scavenger than tyrosol, but both fail to affect cell proliferation rates, cell cycle profile or cell apoptosis in human mammary epithelial cells (MCF10A) or breast cancer cells (MDA-MB-231 and MCF7). We found that hydroxytyrosol decreases the intracellular reactive oxygen species (ROS) level in MCF10A cells but not in MCF7 or MDA-MB-231 cells while very high amounts of tyrosol is needed to decrease the ROS level in MCF10A cells. Interestingly, hydroxytyrosol prevents oxidative DNA damage in the three breast cell lines. Therefore, our data suggest that simple phenol hydroxytyrosol could contribute to a lower incidence of breast cancer in populations that consume virgin olive oil due to its antioxidant activity and its protection against oxidative DNA damage in mammary cells.

Phenotypic and genetic characterization of circulating tumor cells by combining immunomagnetic selection and FICTION techniques.

The presence of circulating tumor cells (CTCs) in breast cancer patients has been proven to have clinical relevance. Cytogenetic characterization of these cells could have crucial relevance for targeted cancer therapies. We developed a method that combines an immunomagnetic selection of CTCs from peripheral blood with the fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasm (FICTION) technique. Briefly, peripheral blood (10 ml) from healthy donors was spiked with a predetermined number of human breast cancer cells. Nucleated cells were separated by double density gradient centrifugation of blood samples. Tumor cells (TCs) were immunomagnetically isolated with an anti-cytokeratin antibody and placed onto slides for FICTION analysis. For immunophenotyping and genetic characterization of TCs, a mixture of primary monoclonal anti-pancytokeratin antibodies was used, followed by fluorescent secondary antibodies, and finally hybridized with a TOP2A/HER-2/CEP17 multicolor probe. Our results show that TCs can be efficiently isolated from peripheral blood and characterized by FICTION. Because genetic amplification of TOP2A and ErbB2 (HER-2) in breast cancer correlates with response to anthracyclines and herceptin therapies, respectively, this novel methodology could be useful for a better classification of patients according to the genetic alterations of CTCs and for the application of targeted therapies.

Effects of functional training on pain, leg strength, and balance in women with fibromyalgia

Aim: The aim of this study was to analyze the effect of 18-week functional training (FT) program consisting in two sessions a week of in-water exercise and one of on-land exercise on pain, strength, and balance in women with fibromyalgia. Methods: A sample consisting of 36 fibromyalgia patients was included in the study. The patients were allocated randomly into the experimental group (EG, n = 20), and control group (CG, n = 16). Standardized field-based fitness tests were used to assess muscle strength (30-s chair stand and handgrip strength) and agility/dynamic balance and static balance. Fibromyalgia impact and pain were analyzed by Fibromyalgia Impact Questionnaire (FIQ), tender points (TPs), visual analog scale (VAS). Results: We observed a significant reduction in the FIQ (p = 0.042), the algometer scale of TP (p = 0.008), TP (p < 0.001), and VAS (p < 0.001) in the EG. The EG shows better results in leg strength (p < 0.001), handgrip strength (p = 0.025), agility/dynamic balance (p = 0.032) and balance (p = 0.006). Conclusions: An 18-week intervention consisting in two sessions of in-water exercise and one session of on-land exercise of FT reduces pain and improves functional capacity in FM patients. These results suggested that FT could play an important role in maintaining an independent lifestyle in patients with FM.

Foot strike patterns in veteran runners with respect to the increase of the speed and fatigue

Introduction: In the last few decades the number of participants in the popular endurance races has increased,especially for athletes in the category of veterans (over 35 years), as well as the number of organized events. Objective: To analyze the foot strike patterns in veteran runners with respect to an increase of speed and fatigue. Methods: Participation of 43 athletes, 40 men (mean age: 41.3±1.10 years old) and 3 women (mean age: 44.3±3.17 years old). After warm-up, three running protocols were performed: comfortable pace, competition pace and competicion pace until the increase two values of the Borg scale, freely chosen by each athlete. Foot strike patterns (contact surface, lateral rotation and tilting of the foot) and a time sequence were analyzed by high-speed video. Results: 84.5% of the athletes had rearfoot, increasing with the speed and fatigue. Among 52.7% of the athletes had mild external rotation and 69% had mild lateral tilt. No relationship was found between athletes, either with or without lesions in the foot strike patterns on the three race protocols. The contact time was higher in rearfoot regarding the rest of the types of foot strike patterns in comfortable running and competition pace. Conclusion: The findings of this study may be relevant for the design of athletic shoes and contribute information on the relationship between the foot strike patterns of the veteran runner and the risk of injuries.INTRODUCCION: En las ultimas decadas, el numero de participantes en las carreras populares de resistencia ha aumentado, especialmente, de los atletas de la categoria de veteranos (mas de 35 anos), asi como el numero de pruebas organizadas.OBJETIVO: Analizar la dinamica de apoyo en la carrera del atleta veterano con relacion al incremento de la velocidad y de la fatiga.METODOS: Participaron 43 atletas, 40 hombres (promedio de edad: 41,3 ± 1,10 anos) y 3 mujeres (promedio de edad 44,3 ± 3,17 anos). Despues del calentamiento, fueron realizados tres protocolos de carrera: confortable, ritmo de competicion y ritmo de competicion hasta el incremento de los valores de la escala de Borg, escogidos libremente por cada atleta. La dinamica de apoyo (superficie de contacto, rotacion lateral e inclinacion del pie) y su secuencia temporal fueron analizados en video de alta velocidad.RESULTADOS: Del total, 84,5% de los atletas presentaron apoyo atrasado, con incremento correspondiente al aumento de la velocidad y de la fatiga en la carrera. De los atletas, 52,7% presentaron rotacion externa leve y 69% presentaron inclinacion lateral leve. No fue encontrada relacion entre atletas lesionados o no ni entre la dinamica de apoyo en los tres tipos de carrera. El tiempo de contacto en la fase de amortiguacion fue superior en el apoyo atrasado con relacion al resto de los tipos de pasos en la carrera confortable y en el ritmo de competicion.CONCLUSION: Los descubrimientos de este estudio pueden ser importantes para el diseno de zapatos deportivos y para aportar informaciones sobre la relacion entre el estandar del paso del corredor veterano y el riesgo de lesiones.

Dinámica de apoyo en corredores veteranos con relación al incremento de la velocidad y de la fatiga

Introduction: In the last few decades the number of participants in the popular endurance races has increased,especially for athletes in the category of veterans (over 35 years), as well as the number of organized events. Objective: To analyze the foot strike patterns in veteran runners with respect to an increase of speed and fatigue. Methods: Participation of 43 athletes, 40 men (mean age: 41.3±1.10 years old) and 3 women (mean age: 44.3±3.17 years old). After warm-up, three running protocols were performed: comfortable pace, competition pace and competicion pace until the increase two values of the Borg scale, freely chosen by each athlete. Foot strike patterns (contact surface, lateral rotation and tilting of the foot) and a time sequence were analyzed by high-speed video. Results: 84.5% of the athletes had rearfoot, increasing with the speed and fatigue. Among 52.7% of the athletes had mild external rotation and 69% had mild lateral tilt. No relationship was found between athletes, either with or without lesions in the foot strike patterns on the three race protocols. The contact time was higher in rearfoot regarding the rest of the types of foot strike patterns in comfortable running and competition pace. Conclusion: The findings of this study may be relevant for the design of athletic shoes and contribute information on the relationship between the foot strike patterns of the veteran runner and the risk of injuries.INTRODUCCION: En las ultimas decadas, el numero de participantes en las carreras populares de resistencia ha aumentado, especialmente, de los atletas de la categoria de veteranos (mas de 35 anos), asi como el numero de pruebas organizadas.OBJETIVO: Analizar la dinamica de apoyo en la carrera del atleta veterano con relacion al incremento de la velocidad y de la fatiga.METODOS: Participaron 43 atletas, 40 hombres (promedio de edad: 41,3 ± 1,10 anos) y 3 mujeres (promedio de edad 44,3 ± 3,17 anos). Despues del calentamiento, fueron realizados tres protocolos de carrera: confortable, ritmo de competicion y ritmo de competicion hasta el incremento de los valores de la escala de Borg, escogidos libremente por cada atleta. La dinamica de apoyo (superficie de contacto, rotacion lateral e inclinacion del pie) y su secuencia temporal fueron analizados en video de alta velocidad.RESULTADOS: Del total, 84,5% de los atletas presentaron apoyo atrasado, con incremento correspondiente al aumento de la velocidad y de la fatiga en la carrera. De los atletas, 52,7% presentaron rotacion externa leve y 69% presentaron inclinacion lateral leve. No fue encontrada relacion entre atletas lesionados o no ni entre la dinamica de apoyo en los tres tipos de carrera. El tiempo de contacto en la fase de amortiguacion fue superior en el apoyo atrasado con relacion al resto de los tipos de pasos en la carrera confortable y en el ritmo de competicion.CONCLUSION: Los descubrimientos de este estudio pueden ser importantes para el diseno de zapatos deportivos y para aportar informaciones sobre la relacion entre el estandar del paso del corredor veterano y el riesgo de lesiones.

Dinâmica do apoio em corredores veteranos com relação ao incremento da velocidade e da fadiga

Introduction: In the last few decades the number of participants in the popular endurance races has increased,especially for athletes in the category of veterans (over 35 years), as well as the number of organized events. Objective: To analyze the foot strike patterns in veteran runners with respect to an increase of speed and fatigue. Methods: Participation of 43 athletes, 40 men (mean age: 41.3±1.10 years old) and 3 women (mean age: 44.3±3.17 years old). After warm-up, three running protocols were performed: comfortable pace, competition pace and competicion pace until the increase two values of the Borg scale, freely chosen by each athlete. Foot strike patterns (contact surface, lateral rotation and tilting of the foot) and a time sequence were analyzed by high-speed video. Results: 84.5% of the athletes had rearfoot, increasing with the speed and fatigue. Among 52.7% of the athletes had mild external rotation and 69% had mild lateral tilt. No relationship was found between athletes, either with or without lesions in the foot strike patterns on the three race protocols. The contact time was higher in rearfoot regarding the rest of the types of foot strike patterns in comfortable running and competition pace. Conclusion: The findings of this study may be relevant for the design of athletic shoes and contribute information on the relationship between the foot strike patterns of the veteran runner and the risk of injuries.INTRODUCCION: En las ultimas decadas, el numero de participantes en las carreras populares de resistencia ha aumentado, especialmente, de los atletas de la categoria de veteranos (mas de 35 anos), asi como el numero de pruebas organizadas.OBJETIVO: Analizar la dinamica de apoyo en la carrera del atleta veterano con relacion al incremento de la velocidad y de la fatiga.METODOS: Participaron 43 atletas, 40 hombres (promedio de edad: 41,3 ± 1,10 anos) y 3 mujeres (promedio de edad 44,3 ± 3,17 anos). Despues del calentamiento, fueron realizados tres protocolos de carrera: confortable, ritmo de competicion y ritmo de competicion hasta el incremento de los valores de la escala de Borg, escogidos libremente por cada atleta. La dinamica de apoyo (superficie de contacto, rotacion lateral e inclinacion del pie) y su secuencia temporal fueron analizados en video de alta velocidad.RESULTADOS: Del total, 84,5% de los atletas presentaron apoyo atrasado, con incremento correspondiente al aumento de la velocidad y de la fatiga en la carrera. De los atletas, 52,7% presentaron rotacion externa leve y 69% presentaron inclinacion lateral leve. No fue encontrada relacion entre atletas lesionados o no ni entre la dinamica de apoyo en los tres tipos de carrera. El tiempo de contacto en la fase de amortiguacion fue superior en el apoyo atrasado con relacion al resto de los tipos de pasos en la carrera confortable y en el ritmo de competicion.CONCLUSION: Los descubrimientos de este estudio pueden ser importantes para el diseno de zapatos deportivos y para aportar informaciones sobre la relacion entre el estandar del paso del corredor veterano y el riesgo de lesiones.