Ca2+ signals in human umbilical endothelial cells derived from pregnancy with fetal growth restriction associated with hypertensive disorder.

Fetal growth restriction associated with hypertensive disorders of pregnancy (FGR-HDP) is a prevalent pathology with a higher risk of perinatal morbimortality. In this condition, placental insufficiency and endothelial dysfunction serve key roles. The present prospective cohort study monitored 11 patients with an FGR-HDP and 15 with full-term normotensive pregnancies and studied post-natal intracellular calcium concentration ([Ca2+]i) signals in human umbilical vein endothelial cells (HUVECs). Small fetuses with placental insufficiency were identified using fetal biometry with Doppler velocimetry. Mean gestational age and birth weight were 31.8±4.1 weeks and 1,260±646 g for FGR-HDP and 39.2±0.8 weeks and 3,320±336 g for normal births, respectively. Abnormal umbilical artery Doppler waveforms were found in 64% of neonates with FGR-HDP. A significant percentage (86%) of FGR newborns were admitted to the neonatal intensive care unit at Gustavo Fricke hospital, Viña del Mar, Chile, with one case of death after birth. [Ca2+]i signals were measured by microfluorimetry in Fluo-3-loaded HUVECs from primary cultures. Altered [Ca2+]i signals were observed in HUVECs from FGR-HDP, where the sustained phase of ATP-induced [Ca2+]i responses was significantly reduced compared with the normotensive group. Also, the [Ca2+]i signals induced with 10 mM Ca2+ after depletion of internal Ca2+ stores were significantly higher. The present study provides a better comprehension of the role of altered cytosolic Ca2+ dynamics in endothelial dysfunction and an in vitro model to assess novel therapeutic approaches for decreasing or preventing complications in FGR-HDP.

Diabetic concentrations of metformin inhibit platelet-mediated ovarian cancer cell progression.

Clinical studies have suggested a survival benefit in ovarian cancer patients with type 2 diabetes mellitus taking metformin, however the mechanism by which diabetic concentrations of metformin could deliver this effect is still poorly understood. Platelets not only represent an important reservoir of growth factors and angiogenic regulators, they are also known to participate in the tumor microenvironment implicated in tumor growth and dissemination. Herein, we investigated if diabetic concentrations of metformin could impinge upon the previously reported observation that platelet induces an increase in the tube forming capacity of endothelial cells (angiogenesis) and upon ovarian cancer cell aggressiveness. We demonstrate that metformin inhibits the increase in angiogenesis brought about by platelets in a mechanism that did not alter endothelial cell migration. In ovarian cancer cell lines and primary cultured cancer cells isolated from the ascitic fluid of ovarian cancer patients, we assessed the effect of combinations of platelets and metformin upon angiogenesis, migration, invasion and cancer sphere formation. The enhancement of each of these parameters by platelets was abrogated by the present of metformin in the vast majority of cancer cell cultures tested. Neither metformin nor platelets altered proliferation; however, metformin inhibited the increase in phosphorylation of focal adhesion kinase induced by platelets. We present the first evidence suggesting that concentrations of metformin present in diabetic patients may reduce the actions of platelets upon both endothelial cells and cancer cell survival and dissemination.

Platelets enhance tissue factor protein and metastasis initiating cell markers, and act as chemoattractants increasing the migration of ovarian cancer cells.

BACKGROUND: An increase in circulating platelets, or thrombocytosis, is recognized as an independent risk factor of bad prognosis and metastasis in patients with ovarian cancer; however the complex role of platelets in tumor progression has not been fully elucidated. Platelet activation has been associated with an epithelial to mesenchymal transition (EMT), while Tissue Factor (TF) protein expression by cancer cells has been shown to correlate with hypercoagulable state and metastasis. The aim of this work was to determine the effect of platelet-cancer cell interaction on TF and "Metastasis Initiating Cell (MIC)" marker levels and migration in ovarian cancer cell lines and cancer cells isolated from the ascetic fluid of ovarian cancer patients. METHODS: With informed patient consent, ascitic fluid isolated ovarian cancer cells, cell lines and ovarian cancer spheres were co-cultivated with human platelets. TF, EMT and stem cell marker levels were determined by Western blotting, flow cytometry and RT-PCR. Cancer cell migration was determined by Boyden chambers and the scratch assay. RESULTS: The co-culture of patient-derived ovarian cancer cells with platelets causes: 1) a phenotypic change in cancer cells, 2) chemoattraction and cancer cell migration, 3) induced MIC markers (EMT/stemness), 3) increased sphere formation and 4) increased TF protein levels and activity. CONCLUSIONS: We present the first evidence that platelets act as chemoattractants to cancer cells. Furthermore, platelets promote the formation of ovarian cancer spheres that express MIC markers and the metastatic protein TF. Our results suggest that platelet-cancer cell interaction plays a role in the formation of metastatic foci.

Metformin, at concentrations corresponding to the treatment of diabetes, potentiates the cytotoxic effects of carboplatin in cultures of ovarian cancer cells.

The use of the type 2 diabetics drug metformin has been correlated with enhanced progression-free survival in ovarian cancer. The literature has speculated that this enhancement is due to the high concentration of metformin directly causing cancer cell death. However, this explanation does not fit with clinical data reporting that the women exposed to constant micromolar concentrations of metformin, as present in the treatment of diabetes, respond better to chemotherapy. Herein, our aim was to examine whether micromolar concentrations of metformin alone could bring about cancer cell death and whether micromolar metformin could increase the cytotoxic effect of commonly used chemotherapies in A2780 and SKOV3 cell lines and primary cultured cancer cells isolated from the peritoneal fluid of patients with advanced ovarian cancer. Our results in cell lines demonstrate that no significant loss of viability or change in cell cycle was observed with micromolar metformin alone; however, we observed cytotoxicity with micromolar metformin in combination with chemotherapy at concentrations where the chemotherapy alone produced no loss in viability. We demonstrate that previous exposure and maintenance of metformin in conjunction with carboplatin produces a synergistic enhancement in cytotoxicity of A2780 and SKOV3 cells (55% and 43%, respectively). Furthermore, in 5 (44%) of the 11 ovarian cancer primary cultures, micromolar metformin improved the cytotoxic response to carboplatin but not paclitaxel or doxorubicin. In conclusion, we present data that support the need for a clinical study to evaluate the adjuvant maintenance or prescription of currently approved doses of metformin during the chemotherapeutic treatment of ovarian cancer.

Paclitaxel-PHBV nanoparticles and their toxicity to endometrial and primary ovarian cancer cells.

This report is an integrated study to include the molecular simulation, physicochemical characterization and biological analysis of a paclitaxel-loaded PHBV nanoparticle that demonstrates uptake, release and cytotoxicity in cancer cell lines. Taking this nanoparticle one step closer to its use in a clinical setting, we demonstrate that it causes significant cell death in primary cultures of stage IIIc serous ovarian cancer cells isolated from six patients. Molecular simulations revealed a high affinity of paclitaxel for the water-polymer interface, thus the drug is delivered only when the polymer near it is degraded. The Fourier transform infrared spectroscopy suggests the formation of a short-lived crystalline phase, also observed in the CG simulations, and transmission electron microscopy revealed branched structures on the surface of particles, which disappeared after 4 days. Biological analyses indicated that these particles have a 48-h window of toxicity protection, allowing for the endocytosis of the particle by the cells; this finding was corroborated by confocal microscopy and flow cytometry. The low cost to synthesize PHBV using microorganisms and the potential chemical modifications of the polymer make it attractive for inexpensive, large-scale pharmaceutical production.

Radical vaginal trachelectomy and laparoscopic pelvic lymphadenectomy in IB1 cervical cancer during pregnancy.

► We expose an novel surgical technique in pregnant women with cervical cancer. ► We preserve the pregnancy in this case. ► After 40 months of monitoring both patient and her child are healthy.

[Molecular epidemiology of a streptococcus pyogenes related nosocomial outbreak in a burn unit]. / Epidemiología molecular de un brote de infecciones por streptococcus pyogenes en una unidad de quemados.

BACKGROUND: Group A Streptococcal (GAS) infections have increased in frequency and severity worldwide. During April 1996, a nosocomial outbreak associated to GAS infections affected seven patients admitted to a pediatric burn unit. The causative organism was likely disseminated from the source patient to another child in the emergency room before he was transferred to the burn unit. Patients developed burn infections or invasive disease. One of them died due to a toxic shock syndrome and 3 other lost their skin grafts. Perineal and nasal microbiological surveillance of 42 related health care workers identified only one of them as carrier of S pyogenes. AIM: To report a molecular analysis of an apparently clonal outbreak. MATERIAL AND METHODS: The available isolates were analyzed by molecular methods including random amplified polymorphic DNA analysis (RAPD) with 4 different primers, Sma-I pulsed field gel electrophoresis (PFGE) analysis, and speA, speB and speC detection by polymerase chain reaction (PCR). RESULTS: Two phylogenetically distant and sequentially isolated bacterial groups were identified either by RAPD analysis with selected primers or by Smal-PFGE analysis. The first group involved isolates identified in two patients that included the lethal case. The second bacterial group comprised 5 clinical isolates and the perineal and nasal isolates obtained from a health care worker. Only strains belonging to the first group harbored the speA gene and were associated with invasive disease. The second group could be split further in two subgroups according to their speB profile. CONCLUSIONS: RAPD analysis with selected primers can reproduce the PFGE-discriminating ability on the epidemiological analysis of GAS infections.

Enfermedad hemolítica del recién nacido (EHRN) por anticuerpos anti-M / Hemolytic disease of the newborn infant caused by anti M antibody

Se describe un caso de enfermedad hemolítica del recién nacido producida por un anticuerpo anti-M. En la paciente se encontraron dos anticuerpos de esta especificidad, uno de los cuales se mostraba como IgM y el otro como IgG. De los glóbulos rojos del recién nacido pudo eluirse el anti-M de tipo IgG y se encontró además un test de Coombs directo positivo. En ambos casos se utilizó sangre de cordón. No pudo realizarse control postparto, ya que la paciente no concurrió

Frecuencia de grupos sanguíneos: ABO, sistema Rh-Hr y factor Du / Frequency of blood groups: ABO, Rh-Hr and Du factor

Este trabajo muestra la incidencia de grupos sanguíneos y del sistema Rh-Hr en una Maternidad Universitaria durante un período de 3 años. Los resultados arrojan para el "O" (55,5%), "A" (33,1%), "B" (8,7%) y "AB" (2,5%), Rho positivos (86,6%), Rho negativos (13,1%) y Du positivos (0,25%)

Enfermedad hemolítica del recién nacido (EHRN) por anticuerpos anti-M / Hemolytic disease of the newborn infant caused by anti M antibody

Se describe un caso de enfermedad hemolítica del recién nacido producida por un anticuerpo anti-M. En la paciente se encontraron dos anticuerpos de esta especificidad, uno de los cuales se mostraba como IgM y el otro como IgG. De los glóbulos rojos del recién nacido pudo eluirse el anti-M de tipo IgG y se encontró además un test de Coombs directo positivo. En ambos casos se utilizó sangre de cordón. No pudo realizarse control postparto, ya que la paciente no concurrió (AU)

Frecuencia de grupos sanguíneos: ABO, sistema Rh-Hr y factor Du / Frequency of blood groups: ABO, Rh-Hr and Du factor

Este trabajo muestra la incidencia de grupos sanguíneos y del sistema Rh-Hr en una Maternidad Universitaria durante un período de 3 años. Los resultados arrojan para el "O" (55,5%), "A" (33,1%), "B" (8,7%) y "AB" (2,5%), Rho positivos (86,6%), Rho negativos (13,1%) y Du positivos (0,25%) (AU)