Symmetric and dissymmetric carbohydrate-phenyl ditriazole derivatives as DNA G-quadruplex ligands: Synthesis, biophysical studies and antiproliferative activity.

Here we present a novel G4-binding family of compounds based on a central core of phenyl ditriazole (PDTZ) modified with carbohydrates and phenyl pyrrolidinyl side-chains. Their synthesis was achieved using controlled click chemistry conditions to obtain both, symmetric and dissymmetric carb-PDTZ derivatives without any intermediate protecting steps through an optimized methodology. Binding of the new carb-PDTZ to a variety of G-quadruplex motifs was examined using different biophysical techniques. The symmetric carb-PDTZ derivatives were not able to stabilize G4, but the dissymmetric ones (containing one sugar and one phenyl pyrrolidinyl side-chain) did. Interestingly, the dissymmetric carb-PDTZ derivatives showed much higher G4 vs duplex DNA selectivity than the control compound PDTZ 1, which contains two phenyl pyrrodilinyl side-chains and no carbohydrates. Their potential antitumoral activity was also investigated by in vitro cytotoxicity measurements on different cancerous cell lines. All carb-PDTZ derivatives showed higher IC50 values than the control PDTZ 1, probably due to the lack of compound stability of some derivatives and to lower cellular uptake.

Oxidative stress and cell death induction by amitraz and its metabolite BTS-27271 mediated through cytochrome P450 and NRF2 pathway alteration in primary hippocampal cell.

Amitraz is a neurotoxic formamidine pesticide that induces cell death in hippocampal neurons, although its mechanisms are unknown. Amitraz produces reactive oxygen species (ROS), which could lead to cell death. Amitraz was shown to induce different cytochrome P450 (CYP) isoenzymes involved with ROS and apoptotic cell death induction. Finally, amitraz was described to decrease the activity of antioxidant enzymes regulated through KEAP1/NRF2 pathway, thus likely leading to a reduction of ROS elimination and to cell death induction. We evaluated the effect of amitraz or BTS-27271 co-treatment with or without the antioxidant N-acetylcysteine and/or the unspecific CYP inhibitor 1-aminobenzotriazole on cell viability and its related mechanisms in wild type and silenced primary hippocampal neurons after 24 h treatment. We observed that amitraz produced oxidative stress and CYPs induction leading to apoptotic cell death. ROS generation was partially mediated by CYPs induction and downregulation of NRF2-pathway through KEAP1 overexpression. These data could help explain the mechanism by which amitraz induces cell death and oxidative stress and provide a therapeutic strategy to protect against this effect in case of poisoning.

G-Quadruplex Identification in the Genome of Protozoan Parasites Points to Naphthalene Diimide Ligands as New Antiparasitic Agents.

G-quadruplexes (G4) are DNA secondary structures that take part in the regulation of gene expression. Putative G4 forming sequences (PQS) have been reported in mammals, yeast, bacteria, and viruses. Here, we present PQS searches on the genomes of T. brucei, L. major, and P. falciparum. We found telomeric sequences and new PQS motifs. Biophysical experiments showed that EBR1, a 29 nucleotide long highly repeated PQS in T. brucei, forms a stable G4 structure. G4 ligands based on carbohydrate conjugated naphthalene diimides (carb-NDIs) that bind G4's including hTel could bind EBR1 with selectivity versus dsDNA. These ligands showed important antiparasitic activity. IC50 values were in the nanomolar range against T. brucei with high selectivity against MRC-5 human cells. Confocal microscopy confirmed these ligands localize in the nucleus and kinetoplast of T. brucei suggesting they can reach their potential G4 targets. Cytotoxicity and zebrafish toxicity studies revealed sugar conjugation reduces intrinsic toxicity of NDIs.

Percepciones de gestores de salud y facilitadores de la estrategia Atención Integrada a las Enfermedades Prevalentes (AIEPI) en una zona del Noreste de Brasil y Perú / Perceptions of health managers and facilitators of the Integrated management of childhood illness (IMCI) in Northeastern Brazil and Peru / Percepções de gestores de saúde e facilitadores da estratégia Atenção Integrada às Doenças Prevalentes na Infância (AIDPI) em uma área do Nordeste do Brasil e Peru

El objetivo del estudio fue explorar las percepciones sobre la estrategia de Atención Integrada a las Enfermedades Prevalentes en la Infancia (AIEPI) de gestores de salud y facilitadores de la estrategia. Se realizó una investigación de tipo cualitativo mediante entrevistas semiestructuradas. Se llevaron a cabo 14 entrevistas entre mayo y junio del 2011, ocho en el Estado de Ceará-Brasil y seis en el Perú. En general los profesionales manifestaron una buena percepción de la estrategia AIEPI, observándose una predominancia del componente clínico, en comparación con el componente comunitario y de servicios de salud. A diferencia de Brasil, en el Perú hubo una promoción gubernamental para la adopción estrategia. En conclusión, este estudio muestra una intervención compleja de la estrategia AIEPI, con predominancia del componente clínico poniendo en evidencia las limitaciones en la integración de los componentes de la estrategia.

The objective of this study was to explore the Integrated Management of Childhood Illness (IMCI) strategy through the perceptions of the health managers and facilitators of strategy. A qualitative study was conducted using semi-structured interviews. Fourteen interviews were carried out, eight in the State of Ceará-Brazil and six in Peru, between May and June 2011. In general, participants expressed a positive perception of the IMCI strategy. Also was observed the predominance of the clinical component, compared with the community or the health services component. In Peru unlike Brazil, the government promoted the adoption of the strategy. In conclusion, this study shows a complex intervention of the IMCI strategy, with a predominance of the clinical component, evidencing the limitations in the integration of the components of the strategy.

O objetivo do estudo foi explorar as percepções da estratégia de Atenção Integrada às Doenças Prevalentes na Infância (AIDPI) de gestores de saúde e facilitadores da estratégia. A pesquisa qualitativa foi realizada por meio de entrevistas semiestruturadas. Foram realizadas 14 entrevistas, entre maio e junho de 2011, oito no Estado do Ceará-Brasil e seis no Peru. De um modo geral, os profissionais relataram uma boa percepção da estratégia AIDPI. Observou-se a predominância do componente clínico, em comparação com os serviços de componentes da comunidade e de saúde. Ao contrário do Brasil, no Peru, houve uma promoção do governo para adoção da estratégia. Em conclusão, este estudo mostra uma intervenção complexa da estratégia AIDPI, com predomínio do componente clínico, colocando em evidência as limitações na integração dos componentes da estratégia.

Amitraz changes NE, DA and 5-HT biosynthesis and metabolism mediated by alterations in estradiol content in CNS of male rats.

Amitraz is a formamidine insecticide/acaricide that alters different neurotransmitters levels, among other neurotoxic effects. Oral amitraz exposure (20, 50 and 80 mg/kg bw, 5 days) has been reported to increase serotonin (5-HT), norepinephrine (NE) and dopamine (DA) content and to decrease their metabolites and turnover rates in the male rat brain, particularly in the striatum, prefrontal cortex, and hippocampus. However, the mechanisms by which these alterations are produced are not completely understood. One possibility is that amitraz monoamine oxidase (MAO) inhibition could mediate these effects. Alternatively, it alters serum concentrations of sex steroids that regulate the enzymes responsible for these neurotransmitters synthesis and metabolism. Thus, alterations in sex steroids in the brain could also mediate the observed effects. To test these hypothesis regarding possible mechanisms, we treated male rats with 20, 50 and 80 mg/kg bw for 5 days and then isolated tissue from striatum, prefrontal cortex, and hippocampus. We then measured tissue levels of expression and/or activity of MAO, catechol-O-metyltransferase (COMT), dopamine-ß-hydroxylase (DBH), tyrosine hydroxylase (TH) and tryptophan hydroxylase (TRH) as well as estradiol levels in these regions. Our results show that amitraz did not inhibit MAO activity at these doses, but altered MAO, COMT, DBH, TH and TRH gene expression, as well as TH and TRH activity and estradiol levels. The alteration of these enzymes was partially mediated by dysregulation of estradiol levels. Our present results provide new understanding of the mechanisms contributing to the harmful effects of amitraz.

Synthesis, Binding Properties, and Differences in Cell Uptake of†G-Quadruplex Ligands Based on Carbohydrate Naphthalene Diimide Conjugates.

The G-quadruplexes (G4s) are currently being explored as therapeutic targets in cancer and other pathologies. Six carbohydrate naphthalene diimide conjugates (carb-NDIs) have been synthesized as G4 ligands to investigate their potential selectivity in G4 binding and cell penetration. Carb-NDIs have shown certain selectivity for G4 structures against DNA duplexes, but different sugar moieties do not induce a preference for a specific G4 topology. Interestingly, when monosaccharides were attached through a short ethylene linker to the NDI scaffold, their cellular uptake was two- to threefold more efficient than that when the sugar was directly attached through its anomeric position. Moreover, a correlation between more efficient cell uptake of these carb-NDIs and their higher toxicity in cancerous cell lines has been observed. Carb-NDIs seem to be mainly translocated into cancer cells through glucose transporters (GLUT), of which GLUT4 plays a major role.

Synthesis, RNAi activity and nuclease-resistant properties of apolar carbohydrates siRNA conjugates.

Oligoribonucleotide conjugates carrying apolar carbohydrates at the 5'-end and the corresponding siRNA duplexes have been prepared using phosphoramidite chemistry. All the carbohydrate-siRNA derivatives were compatible with RNA interference machinery if transfected with oligofectamine. In the absence of a transfection agent, some of them exerted certain reduction of gene expression. Double-tailed permethylated glucose conjugated to siRNA through a long spacer inhibited gene expression up to 26% compared to the scrambled duplex. Such modifications contribute positively to the stability of oligoribonucleotides against 5'-exonuclease degradation.

Ex vivo microbial leakage after using different final irrigation regimens with chlorhexidine.

OBJECTIVE: To assess the influence of final irrigation protocols with chlorhexidine in the coronal leakage of Enterococcus faecalis in filled root canals. MATERIAL AND METHODS: Seventy single-root canals from extracted teeth were prepared using ProTaper instruments. The irrigation protocol accomplished an alternating irrigation with 5 mL of 2.5% sodium hypochlorite (NaOCI) and 17% EDTA between each file. The teeth were randomly divided into four experimental groups (n=15) according to the final irrigation regimen: group 1, without final irrigation; group 2, irrigation with 10 mL 2.0% chlorhexidine (CHX); group 3, with a final application of EC40™; and group 4, irrigation with the combination (1:1) of 0.2% CHX + 0.1% cetrimide (CTR). All the teeth were mounted in a two-chamber apparatus and the coronal access was exposed to E. faecalis. The presence of turbidity in the BHI broth over a period of 180 days was observed. The Friedman test was used for statistical analysis. RESULTS: EC40™ varnish showed the least leakage at 180 days, and was statistically similar to 2% CHX. No significant differences were observed between the group without final irrigation and the 2% CHX group or 0.2% CHX + 0.1% CTR. CONCLUSIONS: In this ex vivo study, EC40™ showed the longest delayed coronal leakage of E. faecalis, although without significant differences from 2% CHX.

Prospective Open-Label, Single-Arm, Multicenter, Safety, Tolerability, and Pharmacokinetic Studies of Intravenous Levetiracetam in Children With Epilepsy.

Levetiracetam given via intravenous administration has been shown to be an effective alternative in adults with epilepsy when oral administration is not feasible. This study was a prospective single-arm, multicenter study to assess tolerability, safety, and pharmacokinetics of intravenous levetiracetam in children with epilepsy. Children with epilepsy ages 1 month to 16 years requiring intravenous levetiracetam were enrolled. Assessments included vital signs, electrocardiogram, hematology, chemistry, plasma concentrations of antiepileptic medications, weight, physical/neurological examinations, and pharmacokinetics. A total of 52 patients were enrolled. Mild to moderate treatment-emergent adverse events occurred in 63%, the most frequent being pyrexia and dry mouth. Most other treatment-emergent adverse events were considered unrelated to intravenous levetiracetam administration. Therefore, intravenous levetiracetam in the acute setting was overall well tolerated in children 1 month to 16 years.

Ex vivo study of bacterial coronal leakage in indirect pulp treatment.

OBJECTIVE: The aim of this study was to evaluate, ex vivo, bacterial coronal leakage with different antimicrobial agents applied to the dentine for indirect pulp treatment (IPT). STUDY DESIGN: Sixty extracted teeth were prepared and randomly distributed into 5 groups (n=10): Group 1: no antimicrobial dentine treatment; group 2: 1% chlorhexidine (CHX)+1% thymol varnish (Cervitec); group 3: 2 % CHX solution; group 4: 40% CHX varnish (EC40) and group 5: Clearfil Protect Bond (CPB). Ten teeth served as controls. The teeth were restored using a resin-modified glass ionomer cement (GIC) and then mounted in a two-chamber device. The coronal access was exposed to Streptococcus mutans for 45 days. The appearance of turbidity in the BHI broth of the lower chamber was considered as specimen leakage. RESULTS: Survival analysis, determined by non parametric Kaplan-Meier and log-rank tests, showed that the best results were for groups EC40+GIC and GIC alone; yet there were not statistically significant differences between them. All specimens of CPB+GIC and 2% CHX+GIC, leaked at 45 days. CONCLUSIONS: In IPT the use of GIC without pretreatment of the dentine and pretreatment with 40% CHX varnish resulted in a significant delay of bacterial coronal leakage.

Prevalence of enamel defects in primary and permanent teeth in a group of schoolchildren from Granada (Spain).

OBJECTIVE: The purpose of this study was to determine and compare the prevalence and presentations of developmental defects of the enamel (DDE) in the primary and permanent dentitions of a group of healthy schoolchildren residing in Granada (Spain). STUDY DESIGN: A total of 1,414 healthy schoolchildren were examined using modified DDE criteria for recording enamel defects. RESULTS: The prevalence of DDE of any type was 40.2% in primary dentition and 52% in permanent dentition (p<0.033). Of the 31,820 primary and permanent teeth examined in the study, 699 (4.1%) primary and 1,232 (8.3%) permanent teeth had some form of DDE. Diffuse opacity was the most common type of DDE observed in primary teeth, and demarcated opacity in the permanent teeth. Enamel hypoplasia was the least prevalent defect in both dentition types. CONCLUSIONS: The study population showed a high prevalence of DDE in primary as well as in permanent dentition, reflecting the current increasing trend of this condition, which should be considered as a significant public health problem.

Ex vivo microbial leakage after using different final irrigation regimens with chlorhexidine

Objective: To assess the influence of final irrigation protocols with chlorhexidine in the coronal leakage of Enterococcus faecalis in filled root canals. Material and Methods: Seventy single-root canals from extracted teeth were prepared using ProTaper instruments. The irrigation protocol accomplished an alternating irrigation with 5 mL of 2.5% sodium hypochlorite (NaOCI) and 17% EDTA between each file. The teeth were randomly divided into four experimental groups (n=15) according to the final irrigation regimen: group 1, without final irrigation; group 2, irrigation with 10 mL 2.0% chlorhexidine (CHX); group 3, with a final application of EC40™; and group 4, irrigation with the combination (1:1) of 0.2% CHX + 0.1% cetrimide (CTR). All the teeth were mounted in a two-chamber apparatus and the coronal access was exposed to E. faecalis. The presence of turbidity in the BHI broth over a period of 180 days was observed. The Friedman test was used for statistical analysis. Results: EC40™ varnish showed the least leakage at 180 days, and was statistically similar to 2% CHX. No significant differences were observed between the group without final irrigation and the 2% CHX group or 0.2% CHX + 0.1% CTR. Conclusions: In this ex vivo study, EC40™ showed the longest delayed coronal leakage of E. faecalis, although without significant differences from 2% CHX.

[Clinical care characteristics and factors associated with in-hospital mortality for lung cancer in Spain]. / Características clinicoasistenciales y factores asociados a la mortalidad intrahospitalaria por cáncer de pulmón en España.

BACKGROUND AND OBJECTIVE: Lung cancer causes high morbimortality in Spain and is currently experiencing a significant increase in women. The aim of this study was to describe differential clinical and health care characteristics by sex, as well as factors associated with and geographic differences of in-hospital mortality. MATERIAL AND METHOD: Descriptive study of episodes registered in the National Hospital Discharge Minimum Basic Data Set for admission type and gender in 2005. Two logistic regression models by sex were built in order to explain the individual influence of variables on in-hospital mortality. Using predictive values of the models, standardized mortality rates were calculated to study the variation between Spanish regions. RESULTS: Women presented a lower mean age, smoking habit and in-hospital mortality than men. However, women presented more adenocarcinomas, greater care in high volume centers, more surgery in readmissions and were subjected to chemotherapy more often in new admissions than men. Adenocarcinoma in men and no specific location in women were associated with higher mortality. Smoking habit and lung diagnosis procedures in men, and middle lobe location and bronchoscopy in women were associated with lower mortality. The geographical mortality pattern detected was similar in both sexes only in some regions of Spain. CONCLUSIONS: Differential clinical characteristics, health care and overall results appear to exist depending on individuals' gender. Recognition of these differences are crucial in order to improve the effectiveness and equity of our health care system.

Dentinogenesis imperfecta: the importance of early treatment.

Dentinogenesis imperfecta, also known as hereditary opalescent dentin, is a dentin development disorder with autosomal dominant transmission that affects both the primary and permanent dentition. A case is reported of a family in which the mother and her 6- and 20-year-old children were diagnosed with dentinogenesis imperfecta type II. The mouths of these patients illustrate the progressive deterioration of affected teeth if not adequately treated. The treatment of the 6-year-old son is described, and therapeutic approaches to this disorder in primary and permanent dentition are reviewed. This family exemplifies the need for the earliest possible diagnosis and treatment of dentinogenesis imperfecta to prevent extensive deterioration of the dentition and occlusion.

[Hospital management of pancreatic cancer in Spain and assessment of factors associated with in-hospital mortality]. / Manejo hospitalario del cáncer de páncreas en España y estudio de los factores asociados a la mortalidad intrahospitalaria.

BACKGROUND: Pancreatic cancer is becoming an increasingly important health problem in Spain. This study aimed to analyze the hospital management of this process and the factors associated with mortality by using an administrative data base. MATERIAL AND METHOD: We performed a descriptive study. Socio-demographic, clinical, diagnostic, and therapeutic variables of episodes registered in the national Hospital Discharge Minimum Data Set for 2004 were gathered. Comorbidity was assessed with the Charlson index. A logistic regression model was built to explain the individual influence of variables on in-hospital mortality. Mortality in the 17 autonomous regions of Spain was analyzed by using standardized mortality rates, through predicted mortality obtained from the multivariate model. RESULTS: The mean age was 68 years and men represented 56%. Readmissions accounted for 80% of the cases. The most frequent localization was in the pancreatic head and the most frequently employed procedures were computed tomography and surgery. In-hospital mortality was 25%, was higher in men, and increased with age. Mortality was higher in new admissions than in readmissions. Factors associated with higher mortality in the multivariate analysis were male sex, age, unspecified location or location in the tail, emergency admission, hospital stay, and comorbidity. Observed mortality was higher than expected in the Canary Islands and Madrid and was lower than expected in Catalonia and the Valencian Community. DISCUSSION: Future studies with more detailed information should be performed to allow the factors associated with in-hospital mortality from pancreatic cancer to be confirmed and to clarify the reasons for the geographical differences identified.

[Analysis of in-hospital mortality from coronary artery bypass grafting surgery]. / Análisis de la mortalidad intrahospitalaria de la cirugía de revascularización coronaria.

INTRODUCTION AND OBJECTIVES: Outcomes research and monitoring are of key importance in efforts to improve health care effectiveness and quality. The aim of this study was to describe in-hospital mortality from coronary artery bypass grafting in Spain. Data in an administrative database were used to estimate the statistical performance of two risk-adjustment methods, the Charlson and Ghali indexes. PATIENTS AND METHOD: From the Spanish Hospital Minimum Basic Data Set corresponding to 1997 and 1998 all records which included a code for coronary artery bypass grafting were selected. With in-hospital mortality as the outcome variable, two risk-adjusted logistic multiple regression models were constructed. RESULTS: The database included 13,203 cases, of which 80% were men; mean age was 64.5 years. In-hospital mortality was 7.3%. The figure was significantly higher for women and increased with age. A score of one on the Charlson and Ghali indexes was associated, respectively, with a 23 and 20% increase in the risk of mortality. Probability calculated with the Hosmer-Lemeshow goodness of fit test was 0.765 and 0.965, and the C index was 0.66 and 0.67. Values of Nagelkerke's R2 were 0.051 y 0.058. CONCLUSIONS: In-hospital mortality from coronary artery bypass grafting is much higher in Spain than in other countries. The Minimum Basic Data Set, a low-cost information system that is easy to access, yields interesting and useful information to measure health care quality.

Evaluación de anticuerpos específicos antivirus herpes simple en pacientes con queratoconjuntivitis dendrica viral / Evaluation of herpes simplex virus specific antibodies in patients with dendritic keratoconjunctivitis

Sueros e anticuerpos específicos para herpes simple virus (HSV) de las clases IgG e IgM fueron administrados entre 52 pacientes con queatitis hepética y 66 pacientes de la casa, clínicamente libres de infecciones herpéticas. Nuestros resultados que la demostración de anticuerpos IgG anti-HSV es de poco valor declarativo para la existencia o ausencia de la enfermedad y que la demostración de IgM anti-HSV anticuerpos no está asociada con la actividad de la enfermedad. Además, la titulación siguiente de anticuerpos específicos HSV en el paciente, no demostró ninguna correlación con la evolución clínica de la enfermedad

Evaluación de la respuesta inmune humoral específica en pacientes con infección conjuntival por virus herpes simple (VHS) / Specific inmune response evaluation in patients with herpetic conjuntivitis

Los anticuerpos séricos específicos de los virus simple (VHS) tanto de la clase IgG como IgM fueron cuantificados en un total de 52 pacientes con el diagnóstico de conjuntivitis herpética y en 66 individuos relacionados con los pacientes y clínicamente sanos. Los resultados obtenidos demuestran el pobre valor predictivo de los anticuerpos IgG anti VHS para predecir la presencia de la enfermedad, así como que la presencia de anticuerpos IgM anti VHS no está asociada a la actividad del padecimiento. Más aún, la titulación subsecuente de anticuerpos específicos anti-VHS en los pacientes no mostró correlación con la evolución clínica de l enfermedad