The synthetic molecule stauprimide impairs cell growth and migration in triple-negative breast cancer.

Stauprimide, a semi-synthetic derivative of staurosporine, is known mainly for its potent differentiation-enhancing properties in embryonic stem cells. Here, we studied the effects of stauprimide in cell growth and migration of triple-negative breast cancer cells in vitro, evaluating its potential antitumoral activity in an orthotopic mouse model of breast cancer in vivo. Our results from survival curves, EdU incorporation, cell cycle analysis and annexin-V detection in MDA-MB-231 cells indicated that stauprimide inhibited cell proliferation, arresting cell cycle in G2/M without induction of apoptosis. A decrease in the migratory capability of MDA-MB-231 was also assessed in response to stauprimide. In this work we pointed to a mechanism of action of stauprimide involving the modulation of ERK1/2, Akt and p38 MAPK signalling pathways, and the downregulation of MYC in MDA-MB-231 cells. In addition, orthotopic MDA-MB-231 xenograft and 4T1 syngeneic models suggested an effect of stauprimide in vivo, increasing the necrotic core of tumors and reducing metastasis in lung and liver of mice. Together, our results point to the promising role of stauprimide as a putative therapeutic agent in triple-negative breast cancer.

Semaphorin3B promotes an anti-inflammatory and pro-resolving phenotype in macrophages from rheumatoid arthritis patients in a MerTK-dependent manner.

Previous works from our group show that Semaphorin3B (Sema3B) is reduced in RA and plays a protective role in a mouse arthritis model. In turn, MerTK plays a protective function in murine arthritis models, is expressed by synovial tissue macrophages and is linked to remission in patients with RA. In this study, we examined the role of Sema3B in the phenotypic characteristics of RA macrophages and the implication of MerTK. Peripheral blood monocytes from RA patients were differentiated into IFN-γ (RA MØIFN-γ) or M-CSF (RA MØM-CSF) macrophages and stimulated with LPS, Sema3B or their combination. Alternatively, RA fibroblast like synoviocytes (FLS) were stimulated with RA MØIFN-γ and RA MØM-CSF supernatants. Gene expression was determined by qPCR and protein expression and activation by flow cytometry, ELISA and western blot. Sema3B down-regulated the expression of pro-inflammatory mediators, in both RA MØIFN-γ and RA MØM-CSF. We observed a similar reduction in RA FLS stimulated with the supernatant of Sema3B-treated RA MØIFN-γ and RA MØM-CSF. Sema3B also modulated cell surface markers in macrophages towards an anti-inflammatory phenotype. Besides, MerTK expression and activation was up-regulated by Sema3B, just as GAS6 expression, Resolvin D1 secretion and the phagocytic activity of macrophages. Importantly, the inhibition of MerTK and neuropilins 1 and 2 abrogated the anti-inflammatory effect of Sema3B. Our data demonstrate that Sema3B modulates the macrophage characteristics in RA, inducing a skewing towards an anti-inflammatory/pro-resolving phenotype in a MerTK-dependant manner. Therefore, here we identify a new mechanism supporting the protective role of Sema3B in RA pathogenesis.

YM155 sensitizes ovarian cancer cells to cisplatin inducing apoptosis and tumor regression.

OBJECTIVE: The objective of this study is to chemosensitize ovarian cancer (OVCa) cells to cisplatin (CDDP) using an inhibitor of Survivin, YM155. The efficacy of YM155 in combination with CDDP was determined in vitro, ex vivo and in vivo. METHODS: Human OVCa cell lines A2780p and their cisplatin-resistant derivative A2780cis, were treated with CDDP, YM155, and the combined treatment (YM155+CDDP), and cell viability, mRNA and protein expression levels, cell-cycle distribution, and DNA damage were then evaluated. Furthermore, the efficacy of YM155 combined with CDDP was further examined in established primary cell cultures and xenograft models. RESULTS: The combination of YM155 with CDDP induced G2/M cell cycle arrest and apoptosis, increased DNA damage, and decreased Survivin levels, especially in A2780cis CDDP-resistant cells. Additionally, YM155 in combination with CDDP sensitized primary cell cultures to CDDP. Studies in vivo showed how this combination significantly decreased the tumor size of OVCa xenografts. CONCLUSIONS: Our results demonstrate that in OVCa cells the expression of Survivin did not affect their sensitivity to YM155, suggesting that Survivin was not the only target of YM155. The combination of YM155 with CDDP could be a good option for therapy of CDDP-resistant OVCa, independently of p53 status.

Effect of organic load and nutrient ratio on the operation stability of the moving bed bioreactor for kraft mill wastewater treatment and the incidence of polyhydroxyalkanoate biosynthesis.

This paper studies the effect of organic load rate (OLR) and nutrient ratio on operation stability of the moving bed bioreactor (MBBR) for kraft mill wastewater treatment, analyzing the incidence of polyhydroxyalkanoate (PHA) production. The MBBR operating strategy was to increase OLR from 0.25 ± 0.05 to 2.41 ± 0.19 kg COD m(-3) d(-1) between phases I and IV. The BOD(5):N:P ratio (100:5:1 and 100:1:0.2) was evaluated as an operation strategy for phases IV to V. A stable MBBR operation was found when the OLR was increased during 225 days in five phases. The maximum absolute fluorescence against the proportion of cells accumulating PHA was obtained for an OLR of 2.41 ± 0.19 kg COD m(-3)d(-1) and a BOD(5):N:P relationship of 100:1:0.2. The increase of PHA biosynthesis is due to the increased OLR and is not attributable to the increased cell concentration, which is maintained constant in stationary status during bioreactor biosynthesis.

[Profiles of cell proliferation and apoptosis in the mouse epithelial regeneration model K6b-E6/E7]. / Perfiles de proliferación celular y apoptosis en elmodelo murino de regeneración epitelial K6b-E6/E7.

BACKGROUND: Mammals have limited epithelial regeneration capacity. The K6b-E6/E7 mice model has been described as useful for the study of epithelial regeneration. The objective of this study is to compare the expression of E6/E7 oncogenes with those of cell proliferation and apoptosis during epithelization. The hypothesis of this study is that alterations in cell proliferation and apoptosis in K6b-E6/E7 mice will only occur during epithelization. METHODS: Deep 2 mm punches were performed in the middle of transgenic and control mice's ears. A biopsy was collected from the epithelization zone 72 hours and 2 weeks post-injury. Assays for cell proliferation and apoptosis were carried out by immunohistochemistry and TUNEL techniques, respectively. RT-PCR in situ was performed to compare E6/E7 expressions in the areas studied. RESULTS: Transgenic strain K6b-E6/E7 presented more proliferative cells and less apoptotic cells in epithelizated zones. This effect was limited to suprabasal stratum only, and correlates with E6/E7 oncogenes expression. Two weeks post-injury, cell proliferation and apoptosis were similar in both samples as the E6/E7 expression went down. CONCLUSION: K6b-E6/E7 mouse model is useful for epithelial regeneration. Its mechanisms should be considered for the treatment of deep wounds.

Cuantificación semiautomática de la función ventricular izquierda y derecha en resonancia magnética cardíaca. Estudio preliminar / Semiautomatic quantification of left and right ventricular function in cardiac magnetic resonance imaging: a preliminary study

Objetivos. El objetivo de este estudio es comparar un método de segmentación semiautomático para cuantificar la función de ambos ventrículos en imágenes de resonancia magnética (RM), con el trazado manual de los contornos. Material y métodos. 17 pacientes con diversas enfermedades cardiovasculares fueron examinados con un equipo de RM de 1,5 Tesla (Magnetom Symphony Quantum; maximum gradient, 30 mT/m; slew rate, 125 T/m/s- Siemens Medical Systems, Erlangen, Germany). Se adquirieron, en todos los estudios, imágenes en modo cine, en eje corto (SSFP, 6mm de grosor de corte, en apnea, desde la base al ápex ventricular). Para reducir la interacción del usuario, solo se utilizó una imagen por paciente para iniciar el método semiautomático. Este método se basa en un algoritmo de crecimiento regional y detección de bordes específicamente diseñado. En todos los pacientes, se cuantificaron los volúmenes telediastólico (VTD), telesistólico (VTS) y la fracción de eyección (FE) de ambos ventrículos. Resultados. No se detectaron diferencias estadísticamente significativas en la cuantificación de la función de ambos ventrículos con los métodos de segmentación propuestos (p>0,05). La diferencia de los volúmenes, aunque cercana a la significación, no conlleva un valor clínico relevante. La correlación al estimar la función ventricular izquierda fue excelente (r>0,9), disminuyendo levemente para la función ventricular derecha (r>0,7). Conclusiones. Nuestro método de segmentación semiautomático permite una cuantificación de la función de ambos ventrículos tan exacta como el método convencional(AU)

Objectives. The aim of this study was to compare a semiautomatic segmentation method to quantify the function of both ventricles in magnetic resonance imaging (MRI) with the manual tracing method. Material and methods. We examined 17 patients with diverse cardiovascular diseases on a 1.5 Tesla MRI unit (Magnetom Symphony Quantum; Siemens Medical Systems, Erlangen, Germany) using the following parameters: maximum gradient, 30 mT/m; and slew rate, 125 T/m/s. In all studies, we acquired images in cine mode in the short axis (SSFP, 6mm slice thickness, from the base to the ventricular apex) with breath holding. To reduce the user interaction, we used only one image per patient to initiate the semiautomatic method. The semiautomatic method was based on a specifically designed algorithm of regional growth and border detection. We quantified the end-diastolic volume (EDV), end-systolic volume (ESV), and the ejection fraction (EF) for both ventricles in all patients. Results. No significant differences between the two segmentation techniques were found in the quantification of either ventricle (p>0.05). The difference in the volumes, although nearly significant, are clinically irrelevant. The correlation for the estimation of left ventricular function was excellent (r>0.9), and the correlation for the estimation of right ventricular function was good (r>0.7). Conclusions. Our semiautomatic segmentation method enables the function of both ventricles to be quantified as accurately as the conventional method(AU)

Fibrinogen drives dystrophic muscle fibrosis via a TGFbeta/alternative macrophage activation pathway.

In the fatal degenerative Duchenne muscular dystrophy (DMD), skeletal muscle is progressively replaced by fibrotic tissue. Here, we show that fibrinogen accumulates in dystrophic muscles of DMD patients and mdx mice. Genetic loss or pharmacological depletion of fibrinogen in these mice reduced fibrosis and dystrophy progression. Our results demonstrate that fibrinogen-Mac-1 receptor binding, through induction of IL-1beta, drives the synthesis of transforming growth factor-beta (TGFbeta) by mdx macrophages, which in turn induces collagen production in mdx fibroblasts. Fibrinogen-produced TGFbeta further amplifies collagen accumulation through activation of profibrotic alternatively activated macrophages. Fibrinogen, by engaging its alphavbeta3 receptor on fibroblasts, also directly promotes collagen synthesis. These data unveil a profibrotic role of fibrinogen deposition in muscle dystrophy.

Improved aerobic biodegradation of abietic acid in ECF bleached kraft mill effluent due to biomass adaptation.

Kraft pulp mill effluents contain elevated concentrations of resin acids, chiefly abietic and dehydroabietic acid, and other lipophilic wood constituents. Resin acids, if not efficiently removed during wastewater treatment, can be responsible for chronic toxicity in aquatic systems. The objective of this study was to investigate the biological removal of abietic acid (AbA) during the treatment of elemental chlorine free (ECF) kraft mill effluents in aerobic lagoons and to assess its improvement with time as a result of biomass adaptation. Under these conditions, the average removal efficiencies of AbA and BOD(5) attained in the aerobic lagoon were high and exceeded 80% and 95%, respectively. Microbial inhibition of non-acclimated and acclimated biomass by AbA was not detected in batch bioassays. Kinetic studies showed that the K(s) and V(m) values equalled 76.7 mg AbA/l and 0.011 l/h, respectively, for the non-acclimated biomass, and 1678 mg AbA/l and 0.13 l/h, respectively, for the acclimated biomass.

Proyecto de gestión de cuidados de la Gerencia de Atención Primaria de Mallorca / 17. Sánchez Ruiz-Cabello FJ, Bellón Saameño JA,

En 2001 se inició el proyecto para facilitar a las enfermeras de atención primaria (AP) la adopción de un modelo conceptual (adaptación de M. Teresa Luis, Carmen Fernández y M. Victoria Ramos del modelo de V. Henderson) que permitiese utilizar el proceso de atención de enfermería como método de trabajo. Objetivos. Mejorar la calidad de los cuidados enfermeros profundizando en el conocimiento del modelo así como integrarlo en la práctica. Aplicar criterios de calidad para la evaluación del producto y adaptar la organización. Descripción del proyecto: Primera fase. Actividad formativa dirigida a las responsables de enfermería (RI) del centro de salud (CS) para facilitar su liderazgo en el proyecto. Segunda fase (2002-2003). Las RI realizaron un "proyecto de mejora de su CS" para analizar la situación y determinar cómo facilitar los cambios necesarios en la práctica enfermera teniendo en cuenta el modelo propuesto y seleccionar los centros piloto. Además se formaron 4 grupos, y cada uno realiza una sesión mensual con el objetivo de profundizar en el conocimiento del modelo e introducir elementos para facilitar la adopción. Resultados. El 82 por ciento de los equipos de AP presentaron un "proyecto de mejora", de los que se seleccionó a 3 como centros piloto. Los 4 grupos han realizado 21 sesiones, en las que trabajaron los problemas/diagnósticos de enfermería más frecuentes en AP, así como una propuesta para la adaptación del proceso enfermero al programa informático e-SIAP, y la adaptación al modelo de los programas de salud. Conclusiones. Consideramos que el proyecto de gestión de cuidados está consolidado. Las propuestas de continuidad son editar la documentación trabajada, hacer realidad la informatización en e-SIAP y formar nuevos grupos de trabajo (AU)

Latent membrane protein-1 oncogene deletions in nasopharyngeal carcinoma in Caucasian patients.

OBJECTIVE: The latent membrane protein-1 (LMP-1) is an Epstein-Barr virus (EBV)-transforming protein expressed in nasopharyngeal carcinoma (NPC). A 30-bp deletion in the LMP-1 oncogene has been described in NPC patients from Asia. The purpose of this study was to evaluate the association between NPC and such an EBV deletion in Caucasian patients. MATERIAL AND METHODS: Twenty-seven patients with a diagnosis of NPC were selected. Most of the NPCs were classified as Stages III and IV using the International Union Against Cancer system. Formalin-fixed, paraffin-embedded NPC specimens were found for these cases. Hematoxylin-eosin slides were reviewed and survival analysis was done using the log-rank method. In situ hybridization for EBV-encoded non-polyadenylated RNAs and expression of LMP-1 by means of immunohistochemistry was also performed. Polymerase chain reaction for LMP-1 oncogene analysis was performed to detect the presence of a 30-bp deletion in NPC specimens and EBV-related controls RESULTS: The 30-bp deletion was identified in 67% of NPC cases and in 30% of controls, a statistically significant difference (p = 0.01, chi2 test). LMP-1 deletion was not statistically associated with a worse prognosis in NPC patients (5-year survival: 33% in wild-LMP-1 strains vs 24% in deleted-LMP-1 strains; p = 0.053, log-rank test). CONCLUSION: A 30-bp deletion in the LMP-1 oncogene is present in more than half of Caucasian NPC cases EBV carrying partial deletions in the LMP-1 oncogene may play a role in the pathogenesis of NPC in Caucasian patients.

Occurrence and antimicrobial susceptibility of Ureaplasma parvum (Ureaplasma urealyticum biovar 1) and Ureaplasma urealyticum (Ureaplasma urealyticum biovar 2) from patients with adverse pregnancy outcomes and normal pregnant women.

A recent phylogenetic analysis of Ureaplasma urealyticum resulted in the proposal to divide their 2 biovars into species. We used PCR to compare the distribution of species and the presence of the tet(M) and int-Tn resistance determinants in 63 strains of Ureaplasma spp. isolated from the amniotic fluid of patients with an adverse pregnancy outcome and in 22 strains obtained from the lower genital tract of healthy pregnant women. We also determined the antimicrobial susceptibility of the organisms to erythromycin and tetracycline. U. parvum was the most frequent Ureaplasma species detected in our study. Thus, 50/63 (79.4%) invasive isolates and 17/22 (77.3%) lower genital tract isolates corresponded to U. parvum, whereas 12/63 (19%) invasive isolates and 4/22 (18.2%) non-invasive strains corresponded to U. urealyticum. A mixture of species was found in 2 women. We found no significant differences in the antimicrobial susceptibility of isolates according to species or origin of isolation. Sixty-two strains of Ureaplasma spp. (74.7%) were susceptible to erythromycin, and 21 strains (25.3%) were intermediately susceptible. Sixty-eight isolates (81.9%) were susceptible to tetracycline, 2 strains (2.4%) were intermediate and 13 strains (15.7%) were resistant. DNA sequences related to the tet(M) determinant and the int-Tn gene were found in all tetracycline-resistant isolates.