[Risk factor control, lifestyle intervention, and prescribed medication in a cohort of patients with ischemic heart disease]. / Control sobre factores de riesgo, intervención sobre estilos de vida, y medicación prescrita en una cohorte de pacientes con cardiopatía isquémica.

OBJECTIVE: The aim of this study was to analyze the recommended prevention measures in our health area for patients discharged after a myocardial infarction. METHODS: This was a retrospective descriptive study that selected patients with acute coronary syndrome in our health area in the previous calendar year. Control of the risk factors observed at the time of the coronary event and at 1 year and medication prescribed 1 year after the episode were studied. Variables including age, sex, control of dyslipidemia, hypertension or diabetes mellitus, adherence to treatment and lifestyle habits were analyzed. RESULTS: Risk factor control was insufficient and sometimes even unassessed at the time of infarction. Although a slight improvement was perceived, control remained insufficient 1 year later. Moreover, patients, particularly women, were undertreated: one fifth (20%) more men were receiving appropriate treatment than women year after the myocardial event. CONCLUSIONS: An additional effort must be made compared to what is currently being done, both by specialists in Hospital Care and Primary Care, to carry out good control of risk factors, meaning the control of certain diseases such as diabetes, high blood pressure or dyslipidemia, as well as habits or lifestyles that increase the probability of suffering a cardiovascular event. Furthermore, it is important to avoid these cardiovascular diseases and their relapse to reinforce adherence to the prescribed treatments.

Data on natural radionuclide's activity concentration of cement-based materials.

Cement based materials may contain varying levels of radionuclides, mainly 226Ra (from the 238U series), 232Th and 40K, which are used to determine the Activity Concentration Index ("ACI"). According to the European directive Euratom 2013/59 in these materials, the "ACI" must be < 1 to be suitable for their use in construction. In this paper, data on the activity concentration of natural radionuclides in cement-based materials (i.e. cements, additions, pigments and aggregates) as well as their chemical composition are presented. Radioactivity measurements have been determined by using gamma spectroscopy the chemical compositions have been determined by X-Ray Fluorescence. Data for cements measured shown that white cements present a lower concentration of activity than conventional CEM I. In addition, the CAC (Calcium aluminate cements) present high activity concentration in the 232Th series. Regarding additions, FA (Fly Ash) are those that present the highest concentration of activity in the 238U and 232Th series, while olive biomass ashes are those supplementary cementitious materials that show the highest concentration of activity for 40K. Some pigments used in mortar and concrete technology were also characterized. Granitic and volcanic rocks, potentially used as aggregates present much higher activity concentration than the siliceous aggregate.

Characteristic limits of 230Th in alpha spectrometry with 229Th as tracer, calculated by simulating interfering tails and overlapping peaks.

This study explores the effect of 229Th tracer tail interference on the determination of the sensitivity of 230Th alpha spectrometry of samples with environmental levels of radioactivity. Tracer peak tail interference was calculated with Suma-Alpha, whilst Visual Basic for Applications (VBA in Excel©) software was used to study the variation in sensitivity in terms of the amount of tracer added. Unnecessary increases in the amount of tracer or extended sample measuring times were observed to have adverse effects on method sensitivity (Detection Limit- Ld).

Use of Genie 2000 and Excel VBA to correct for γ-ray interference in the determination of NORM building material activity concentrations.

The γ-radiation emitted by building materials is calculated from the activity indices for 232Th, 226Ra and 40K and expressed as the activity concentration index (ACI). Gamma spectroscopy is a non-destructive technique frequently used to simultaneously determine the indices for several radionuclides. Spectral interpretation poses a number of challenges, including identification of γ-lines subject to summing-in effects, interference from other γ-ray emitting radionuclides and the time required to reach secular equilibrium. These challenges are not fully addressed by Canberra Industries' Genie 2000, the software used by many laboratories to analyse samples. This article describes a Microsoft Excel workbook that exploits Genie 2000 flexibility to program applications with Visual Basic using Canberra's Nuclear Data Access Library and batch procedure tools. The workbook determines 40K activity concentration after correcting for 228Ac interference and 226Ra activity directly from the γ-peak at 186.5 keV. The method proposed was tested by participating in 13 national and international scale inter-comparison exercises. The results were statistically indistinguishable from the reference values at a coverage factor of k = 3 and no statistically significant differences were identified between the respective means by a Student's t pairwise comparison.

Randomised clinical trial: mesalazine versus placebo in the prevention of diverticulitis recurrence.

BACKGROUND: Previous studies have reached conflicting conclusions regarding the efficacy of mesalazine in the prevention of recurrent diverticulitis. AIM: To investigate the efficacy and safety of mesalazine granules in the prevention of recurrence of diverticulitis after acute uncomplicated diverticulitis. METHODS: Two phase 3, randomised, placebo-controlled, double-blind multicentre trials (SAG-37 and SAG-51) investigated mesalazine granules in patients with prior episodes (<6 months) of uncomplicated left-sided diverticulitis. Patients were randomised to receive either 3 g mesalazine once daily or placebo (SAG-37, n=345) or to receive either 1.5 g mesalazine once daily, 3 g once daily or placebo for 96 weeks (SAG-51, n=330). The primary endpoint was the proportion of recurrence-free patients during 48 weeks (SAG-37 and SAG-51) or 96 weeks (SAG-51) of treatment. RESULTS: Mesalazine did not increase the proportion of recurrence-free patients over 48 or 96 weeks compared to placebo. In SAG-37, the proportion of recurrence-free patients during 48 weeks was 67.9% with mesalazine and 74.4% with placebo (P=.226). In SAG-51, the proportion of recurrence-free patients over 48 weeks was 46.0% with 1.5 g mesalazine, 52.0% with 3 g mesalazine and 58.0% with placebo (P=.860 for 3 g mesalazine vs placebo) and over 96 weeks 6.9%, 9.8% and 23.1% respectively (P=.980 for 3 g mesalazine vs placebo). Patients with only one diverticulitis episode in the year prior to study entry had a lower recurrence risk compared to >1 episode. Safety data revealed no new adverse events. CONCLUSION: Mesalazine was not superior to placebo in preventing recurrence of diverticulitis.

The Latin American experience of allografting patients with severe aplastic anaemia: real-world data on the impact of stem cell source and ATG administration in HLA-identical sibling transplants.

We studied 298 patients with severe aplastic anaemia (SAA) allografted in four Latin American countries. The source of cells was bone marrow (BM) in 94 patients and PBSCs in 204 patients. Engraftment failed in 8.1% of recipients with no difference between BM and PBSCs (P=0.08). Incidence of acute GvHD (aGvHD) for BM and PBSCs was 30% vs 32% (P=0.18), and for grades III-IV was 2.6% vs 11.6% (P=0.01). Chronic GvHD (cGvHD) between BM and PBSCs was 37% vs 59% (P=0.002) and extensive 5% vs 23.6% (P=0.01). OS was 74% vs 76% for BM vs PBSCs (P=0.95). Event-free survival was superior in patients conditioned with anti-thymocyte globulin (ATG)-based regimens compared with other regimens (79% vs 61%, P=0.001) as excessive secondary graft failure was seen with other regimens (10% vs 26%, P=0.005) respectively. In multivariate analysis, aGvHD II-IV (hazard ratio (HR) 2.50, confidence interval (CI) 1.1-5.6, P=0.02) and aGvHD III-IV (HR 8.3 CI 3.4-20.2, P<0.001) proved to be independent negative predictors of survival. In conclusion, BM as a source of cells and ATG-based regimens should be standard because of higher GvHD incidence with PBSCs, although the latter combining with ATG in the conditioning regimen could be an option in selected high-risk patients.

Splicing regulator SLU7 preserves survival of hepatocellular carcinoma cells and other solid tumors via oncogenic miR-17-92 cluster expression.

Resisting death is a central hallmark of cancer cells. Tumors rely on a number of genetic mechanisms to avoid apoptosis, and alterations in mRNA alternative splicing are increasingly recognized to have a role in tumorigenesis. In this study, we identify the splicing regulator SLU7 as an essential factor for the preservation of hepatocellular carcinoma (HCC) cells viability. Compared with hepatocytes, SLU7 expression is reduced in HCC cells; however, further SLU7 depletion triggered autophagy-related cellular apoptosis in association with the overproduction of reactive oxygen species. Remarkably, these responses were not observed in primary human hepatocytes or in the well-differentiated HepaRG cell line. Mechanistically, we demonstrate that SLU7 binds the C13orf25 primary transcript in which the polycistronic oncomir miR-17-92 cluster is encompassed, and is necessary for its processing and expression. SLU7 knockdown altered the splicing of the C13orf25 primary transcript, and markedly reduced the expression of its miR-17, miR-20 and miR-92a constituents. This led to the upregulation of CDKN1A (P21) and BCL2L11 (BIM) expression, two bona fide targets of the miR-17-92 cluster and recognized mediators of its pro-survival and tumorigenic activity. Interestingly, altered splicing of miR-17-92 and downregulation of miR-17 and miR-20 were not observed upon SLU7 knockdown in non-transformed hepatocytes, but was found in other (HeLa, H358) but not in all (Caco2) non-hepatic tumor cells. The functional relevance of miR-17-92 dysregulation upon SLU7 knockdown was established when oxidative stress, autophagy and apoptosis were reversed by co-transfection of HCC cells with a miR-17 mimic. Together, these findings indicate that SLU7 is co-opted by HCC cells and other tumor cell types to maintain survival, and identify this splicing regulator as a new determinant for the expression of the oncogenic miR-17-92 cluster. This novel mechanism may be exploited for the development of antitumoral strategies in cancers displaying such SLU7-miR-17-92 crosstalk.

C-Jun N-terminal kinases are required for oncolytic adenovirus-mediated autophagy.

Oncolytic adenoviruses, such as Delta-24-RGD (Δ24RGD), are replication-competent viruses that are genetically engineered to induce selective cancer cell lysis. In cancer cells, Δ24RGD induces massive autophagy, which is required for efficient cell lysis and adenoviral spread. Understanding the cellular mechanisms underlying the regulation of autophagy in cells treated with oncolytic adenoviruses may provide new avenues to improve the therapeutic effect. In this work, we showed that cancer cells infected with Δ24RGDundergo autophagy despite the concurrent activation of the AKT/mTOR pathway. Moreover, adenovirus replication induced sustained activation of JNK proteins in vitro. ERK1/2 phosphorylation remained unchanged during adenoviral infection, suggesting specificity of JNK activation. Using genetic ablation and pharmacological inactivation of JNK, we unequivocally demonstrated that cells infected with Δ24RGD required JNK activation. Thus, genetic co-ablation of JNK1 and JNK2 genes or inhibition of JNK kinase function rendered Δ24RGD-treated cells resistant to autophagy. Accordingly, JNK activation induced phosphorylation of Bcl-2 and prevented the formation of Bcl-2/Beclin 1 autophagy suppressor complexes. Using an orthotopic model of human glioma xenograft, we showed that treatment with Δ24RGD induced phosphorylation and nuclear translocation of JNK, as well as phosphorylation of Bcl-2. Collectively, our data identified JNK proteins as an essential mechanistic link between Δ24RGD infection and autophagy in cancer cells. Activation of JNK without inactivation of the AKT/mTOR pathway constitutes a distinct molecular signature of autophagy regulation that differentiates Δ24RGD adenovirus from the mechanism used by other oncolytic viruses to induce autophagy and provides a new rationale for the combination of oncolytic viruses and chemotherapy.

Vigilancia de infecciones y otros eventos adversos en pacientes en diálisis en el área sur de Gran Canaria / Surveillance for infections and other adverse events in dialysis patients in southern Gran Canaria

Introducción: Las infecciones bacterianas representan un gran desafío en las estrategias de gestión del riesgo, prevención y seguridad del paciente en hemodiálisis de las cuales las infecciones del acceso vascular (AV) representan la primera causa morbi-mortalidad en estos pacientes. Métodos: Estudio prospectivo de incidencia de eventos adversos e infecciones de 7 meses (marzo-septiembre 2008) en las unidades de Hemodiálisis del Área sanitaria Sur de Gran Canaria (Hospital y Centro periférico) utilizando la metodología del Dialysis Surveillance Network del CDC. Resultados: Se vigilaron 1545 pacientes/mes, 60,5% con fístula (FAV), 35,5% con catéter permanente (CP), 3,0% con prótesis y 1,0% con catéter temporal. La incidencia de eventos en ambos centros fue 8,7 casos por 100 pacientes-mes; la tasa de eventos infecciosos fue de 9,1 para FAV y 20,6 para CP en ámbito hospitalario, mientras las tasas de otras infecciones (respiratorias, herida, orina) fueron similares. Se realizó cultivo antes de empezar tratamiento antibiótico en el 91,0% frente a sospecha de bacteriemia y/o infección AV. El 90,0% de tratamientos se ajustaron con antibiograma. Destaca una baja incidencia de bacterias mutirresistentes mientras que las infecciones relacionadas con el AV fueron causadas en proporción similar por bacterias grampositivas y gramnegativas. Conclusiones: El acceso vascular es el principal factor de riesgo para el desarrollo de infecciones. La vigilancia epidemiológica he permitido detectar oportunidades de mejora en ámbitos asistenciales distintos, integrándose como elemento fundamental en el desarrollo de estrategias multidisciplinarias de seguridad del paciente (AU)

Background: Bacterial infections pose a great challenge to risk management activities in the area of chronic haemodialysis, as vascular access related infections are the main cause of mortality among these patients. Methods: Prospective surveillance study lasting 7 months (March-September, 2008) at the two haemodialysis units in a district health area in Gran Canaria, Spain. We have used methodology proposed by CDC´s Dialysis Surveillance Network. Results: 1545 patientsmonth were enrolled, 60,5% having an arterio-venous fistula (AVF), 35,5% permanent catheter (PC), 3,0% graft and 1,0% temporary catheters. Events incidence rate at both centers was 8,6 cases per 100 patients-month, 9,1 rate for FAV and 2,9 rates for CP, So, the greatest incidence of vascular access related infections was for permanent catheter as compared with AFV. Nevertheless the other type of infections (respiratory, urinary tract, skin and chronic ulcers) showed a similar rate. Microbiological cultures before antibiotic treatment were performed in 82,2 %, but increased up to 91,0% when a vascular related infection was suspected. Empiric treatment was adjusted to antibiogram results in 90,0% of occasions. A low incidence of multirresistant microbes was seen. Gram-positive and gram-negative bacteria appeared in a similar proportion. Conclusions: Vascular access is the main risk factor for infectious events. Epidemiological surveillance has allowed us to detect areas of improvement in different settings, appearing as a key element in the risk management and patient safety areas (AU)

Surveillance for infections and other adverse events in dialysis patients in southern Gran Canaria.

BACKGROUND: Bacterial infections pose a major challenge to risk management activities in the area of chronic haemodialysis, as vascular access-related infections are the main cause of mortality among these patients. METHODS: Prospective surveillance study lasting 7 months (March-September, 2008) at two haemodialysis units in a district health area Gran Canaria, Spain. We used the methodology proposed by CDC´s Dialysis Surveillance Network. RESULTS: 1545 patients/month were recorded, 60.5% with an arteriovenous fistula (AVF), 35.5% with a permanent catheter (PC), 3.0% with grafts and 1.0% with temporary catheters. The rate of adverse events was 8.6 cases per 100 patients/month, 9.1 for AVF patients, and 2.9 for PC. Nevertheless, the other types of infections (respiratory, urinary tract, skin and chronic ulcers) showed similar rates. Microbiological cultures were taken in 82.2%, but this rate increased to 91.0% when a vascular access-related infection was suspected. Empirical treatment was adjusted to antibiogram results in 90.0% of occasions. A low incidence of multi-resistant microbes was observed. Gram-positive and gram-negative bacteria appeared in similar proportions. CONCLUSIONS: Vascular access is the main risk factor for infectious events. Epidemiological surveillance has allowed us to detect areas of improvement in different settings, acting as a key element in risk management and patient safety.

PP2A impaired activity is a common event in acute myeloid leukemia and its activation by forskolin has a potent anti-leukemic effect.

Protein phosphatase 2A (PP2A) is a human tumor suppressor that inhibits cellular transformation by regulating the activity of several signaling proteins critical for malignant cell behavior. PP2A has been described as a potential therapeutic target in chronic myeloid leukemia, Philadelphia chromosome-positive acute lymphoblastic leukemia and B-cell chronic lymphocytic leukemia. Here, we show that PP2A inactivation is a recurrent event in acute myeloid leukemia (AML), and that restoration of PP2A phosphatase activity by treatment with forskolin in AML cells blocks proliferation, induces caspase-dependent apoptosis and affects AKT and ERK1/2 activity. Moreover, treatment with forskolin had an additive effect with Idarubicin and Ara-c, drugs used in standard induction therapy in AML patients. Analysis at protein level of the PP2A activation status in a series of patients with AML at diagnosis showed PP2A hyperphosphorylation in 78% of cases (29/37). In addition, we found that either deregulated expression of the endogenous PP2A inhibitors SET or CIP2A, overexpression of SETBP1, or downregulation of some PP2A subunits, might be contributing to PP2A inhibition in AML. In conclusion, our results show that PP2A inhibition is a common event in AML cells and that PP2A activators, such as forskolin or FTY720, could represent potential novel therapeutic targets in AML.

EVI1 controls proliferation in acute myeloid leukaemia through modulation of miR-1-2.

BACKGROUND: The EVI1(ecotropic virus integration site 1) gene codes for a zinc-finger transcription factor, whose transcriptional activation leads to a particularly aggressive form of acute myeloid leukaemia (AML). Although, EVI1 interactions with key proteins in hematopoiesis have been previously described, the precise role of this transcription factor in promoting leukaemic transformation is not completely understood. Recent works have identified specific microRNA (miRNA) signatures in different AML subgroups. However, there is no analysis of miRNAs profiles associated with EVI1 overexpression in humans. METHODS: We performed QT-RT-PCR to assess the expression of 250 miRNAs in cell lines with or without EVI1 overexpression and in patient samples. We used ChIP assays to evaluated the possible binding of EVI1 binding to the putative miRNA promoter. Proliferation of the different cell lines transfected with the anti- or pre-miRs was quantified by MTT. RESULTS: Our data showed that EVI1 expression was significantly correlated with the expression of miR-1-2 and miR-133-a-1 in established cell lines and in patient samples. ChIP assays confirmed that EVI1 binds directly to the promoter of these two miRNAs. However, only miR-1-2 was involved in abnormal proliferation in EVI1 expressing cell lines. CONCLUSIONS: Our data showed that EVI1 controls proliferation in AML through modulation of miR-1-2. This study contributes to further understand the transcriptional networks involving transcription factors and miRNAs in AML.

[Independent multiple primary tumors and second primary neoplasms. Relationship between smoking]. / Tumores primarios múltiples independientes y segundas neoplasias primarias. Relación con el hábito de fumar.

Multiple primary tumors and second primary neoplasms have been increasing in incidence in recent decades and are reviewed in this paper. The reasons attributed to this significant increase are fundamentalment the best diagnosis of multiple concurrent cases and increased overall survival of patients diagnosed with cancer, allowing surface new primary tumors in other organs during or after standard monitoring. At the same time are invoked as possible causes of the widespread use of radio and chemotherapy for the first tumor. The genitourinary system is frequently involved in cases of multiple neoplasms; urological organs are one of the few settlement sites of primary tumors in almost a quarter of cases. This suggests a susceptibility/genitourinary system increased target for neoplastic disease. For this same reason, the urologist has a fundamental role in managing these patients and especially to follow up. We believe that the concept of clinical monitoring of this subset of patients should be revised, and should entail a screening of the most common second primary neoplasms since the risk of developing a subsequent independent cancer after presenting a urothelial tumor is considerably increased.

Tumores primarios múltiples independientes y segundas neoplasias primarias. Relación con el hábito de fumar / Independent multiple primary tumors and second primary neoplasms. Relationship between smoking

Los tumores primarios múltiples, así como las segundas neoplasias primarias, han experimentado un aumento de la incidencia en estas últimas décadas y son objeto de revisión en este trabajo. Los motivos que se atribuyen a este aumento significativo son fundamentalmente el mejor diagnóstico de los casos múltiples concomitantes y la mayor supervivencia en general de los pacientes diagnosticados de cáncer, lo que permite que afloren nuevos tumores primarios en otros órganos durante o después del seguimiento estándar. Al mismo tiempo, se invocan como posibles causas el extenso uso de radioterapia y quimioterapia para el primer tumor. El sistema genitourinario está muy frecuentemente implicado en los casos de neoplasias múltiples; los órganos urológicos son uno de los sitios de asentamiento de algunos de los tumores primarios en casi una cuarta parte de los casos. Esto sugiere una susceptibilidad/diana incrementada del sistema genitourinario para la enfermedad neoplásica. Y, por esta misma razón, el urólogo tiene una responsabilidad esencial en el manejo de estos pacientes y de manera especial durante el seguimiento. Creemos que el concepto de seguimiento clínico de este subgrupo de pacientes debe ser revisado y debe comportar un cribaje de las más frecuentes segundas neoplasias primarias, ya que el riesgo de desarrollar un cáncer independiente subsiguiente después de presentar un tumor urotelial está considerablemente incrementado (AU)

Multiple primary tumors and second primary neoplasms have been increasing in incidence in recent decades and are reviewed in this paper. The reasons attributed to this significant increase are fundamentalment the best diagnosis of multiple concurrent cases and increased overall survival of patients diagnosed with cancer, allowing surface new primary tumors in other organs during or after standard monitoring. At the same time are invoked as possible causes of the widespread use of radio and chemotherapy for the first tumor. The genitourinary system is frequently involved in cases of multiple neoplasms; urological organs are one of the few settlement sites of primary tumors in almost a quarter of cases. This suggests a susceptibility/genitourinary system increased target for neoplastic disease. For this same reason, the urologist has a fundamental role in managing these patients and especially to follow up. We believe that the concept of clinical monitoring of this subset of patients should be revised, and should entail a screening of the most common second primary neoplasms since the risk of developing a subsequent independent cancer after presenting a urothelial tumor is considerably increased (AU)

Tie2-mediated multidrug resistance in malignant gliomas is associated with upregulation of ABC transporters.

Resistance and relapse are still primary causes that result in poor effectiveness of chemotherapy in malignant gliomas. Therefore, development of new therapeutic strategies requires the identification of key molecular pathways regulating chemoresistance. We previously found that abnormal high expression of the Tie2 receptor in gliomas was associated with tumor malignancy. Here, we studied the role of Tie2 activation in drug resistance by testing the cytotoxicity of several chemotherapeutic drugs in a panel of human glioma cell lines and brain tumor stem cells and found that Tie2 activation was significantly related to chemoresistance. The essential role of Tie2 in this phenotype was illustrated by silencing Tie2 using specific siRNA, and the subsequent abrogation of the angiopoietin 1 (Ang1)-mediated chemoresistance. Using quantitative real-time PCR and functional drug efflux studies, we observed that Tie2 activation resulted in increased expression of ATP-binding cassette (ABC) transporters. Consistent with these results, downmodulation of ABCG2 or ABCC2 resulted in the inability of Tie2 activation to induce a chemoresistant phenotype. Our results indicate that Tie2 activation may be important in modifying the evolution of gliomas during conventional chemotherapy regimens, and open new avenues for the search of more effective therapies to avoid the inevitable brain tumor recurrence.

Oncolytic adenovirus retargeted to Delta-EGFR induces selective antiglioma activity.

The fact that glioblastomas, which are one of the most devastating cancers, frequently express the Delta-EGFR (epithelial growth factor receptor) also called mutant variant III of EGFR (EGFRvIII) suggests that this cancer cell-specific receptor might serve as an ideal target for cancer therapy. To assess its potential as such a target, we constructed an oncolytic adenovirus with Retargeted Infectivity Via EGFR (Delta-24-RIVER) on the backbone of Delta-24. This new oncolytic adenovirus targets, as Delta-24 does, the disrupted Rb pathway in cancer cells; in addition, this adenovirus has also been retargeted through the abrogation of CAR binding (Y477A mutation in adenoviral fiber protein) and insertion of an EGFRvIII-specific binding peptide in the HI loop of the fiber protein. As compared with Delta-24, Delta-24-RIVER induced EGFRvIII-selective cytotoxicity in U-87 MG isogenic cell lines and in tetracycline-inducible EGFRVIII expressing U-251 MG cells. Accordingly, by tittering the viral progeny and examining fiber protein expression in the above cells, we showed that the replication of this new construct also correlated with EGFRvIII expression. Consistently, immunohistochemistry staining of the adenoviral capsid protein hexon in the virus-treated tumors revealed that the virus replicated more efficiently in EGFRvIII-expressing U-87 MG.DeltaEGFR xenografts than in the tumors grown from U-87 MG cells. Importantly, treatment with Delta-24-RIVER prolonged the survival of animals with intracranial xenografts derived from U-87 MG.DeltaEGFR cells. Therefore, our results constitute the first proof of the direct targeting of a cancer-specific receptor using an oncolytic adenovirus.

Angiopoietin-2 decreases vascular endothelial growth factor expression by modulating HIF-1 alpha levels in gliomas.

Angiogenesis is thought to depend on a perfectly coordinated balance between endogenous-positive and negative regulatory factors. Of these factors, the vascular endothelial growth factor (VEGF) and angiopoietins (Angs) seem to play an essential role. Recently, we reported the expression of the Ang-natural receptor, Tie2, in neoplastic astrocytic cells within gliomas. Because of the VEGF/Ang2 functional partnership together with the presence of Tie2 in gliomas, we hypothesized a role of Ang2 on the modulation of VEGF levels in these tumors. We examined the effect of Ang2 on VEGF expression in a panel of glioma cells, which showed that Ang2 inhibited VEGF expression at both mRNA and protein levels in Tie2-expressing cells, but not in Tie2-negative cells. VEGF promoter analysis showed that Ang2 regulated VEGF expression at the transcriptional level in relation to a decrease in HIF-1alpha expression and HIF-DNA-binding activity. Tie2 silencing by siRNA rescued the Ang2-mediated downmodulation of VEGF, suggesting an essential role for Tie2 in this regulatory loop. To our knowledge, this is the first report on the role of Ang2/Tie2 in the regulation of HIF-1alpha/VEGF expression, providing additional evidence of the intrinsic coordination that occurs among these factors during angiogenesis.

Combination of the oncolytic adenovirus ICOVIR-5 with chemotherapy provides enhanced anti-glioma effect in vivo.

Novel therapies are clearly needed for gliomas, and the combination of oncolytic vectors with chemotherapy possesses a significant hope for the treatment of this malignancy. In addition, combination with chemotherapy allows for lower virus doses to achieve anticancer effect, thus resulting in lower undesirable toxicities due to viral proteins. In this work, we sought to determine whether combination of an oncolytic adenovirus ICOVIR-5, with RAD001 or temozolomide (TMZ) could result in enhanced anti-glioma effect in vivo. We assessed the in vitro cytotoxic effect and replication properties of ICOVIR-5 in combination with RAD001 or TMZ in U87 MG glioma cell line by MTT and TCID(50), respectively. Our data showed that in vitro treatment with RAD001 or TMZ not only interfered with adenovirus replication but, in addition, enhanced its oncolytic properties. To evaluate the in vivo anticancer effect, athymic mice bearing glioma xenografts (5 x 10(5) U87 MG cells/animal) received a single intratumoral injection of ICOVIR-5 (10(7) PFU/animal). RAD001 was given as a regimen of 5 mg/kg 5 days per week until the end of the experiment and TMZ was administered for 5 days at 7.5 mg/kg/mice. Of significance, combination of ICOVIR-5 with RAD001 or TMZ showed a potent anti-glioma effect in vivo, resulting in a dramatic extension of the median animal survival and in 20-40% animals becoming free of disease beyond 90 days.