Efficient In Vitro and In Vivo Anti-Inflammatory Activity of a Diamine-PEGylated Oleanolic Acid Derivative.

Recent evidence has shown that inflammation can contribute to all tumorigenic states. We have investigated the anti-inflammatory effects of a diamine-PEGylated derivative of oleanolic acid (OADP), in vitro and in vivo with inflammation models. In addition, we have determined the sub-cytotoxic concentrations for anti-inflammatory assays of OADP in RAW 264.7 cells. The inflammatory process began with incubation with lipopolysaccharide (LPS). Nitric oxide production levels were also determined, exceeding 75% inhibition of NO for a concentration of 1 µg/mL of OADP. Cell-cycle analysis showed a reversal of the arrest in the G0/G1 phase in LPS-stimulated RAW 264.7 cells. Furthermore, through Western blot analysis, we have determined the probable molecular mechanism activated by OADP; the inhibition of the expression of cytokines such as TNF-α, IL-1ß, iNOS, and COX-2; and the blocking of p-IκBα production in LPS-stimulated RAW 264.7 cells. Finally, we have analyzed the anti-inflammatory action of OADP in a mouse acute ear edema, in male BL/6J mice treated with OADP and tetradecanoyl phorbol acetate (TPA). Treatment with OADP induced greater suppression of edema and decreased the ear thickness 14% more than diclofenac. The development of new derivatives such as OADP with powerful anti-inflammatory effects could represent an effective therapeutic strategy against inflammation and tumorigenic processes.

On the Use of a Multimodal Optimizer for Fitting Neuron Models. Application to the Cerebellar Granule Cell.

This article extends a recent methodological workflow for creating realistic and computationally efficient neuron models whilst capturing essential aspects of single-neuron dynamics. We overcome the intrinsic limitations of the extant optimization methods by proposing an alternative optimization component based on multimodal algorithms. This approach can natively explore a diverse population of neuron model configurations. In contrast to methods that focus on a single global optimum, the multimodal method allows directly obtaining a set of promising solutions for a single but complex multi-feature objective function. The final sparse population of candidate solutions has to be analyzed and evaluated according to the biological plausibility and their objective to the target features by the expert. In order to illustrate the value of this approach, we base our proposal on the optimization of cerebellar granule cell (GrC) models that replicate the essential properties of the biological cell. Our results show the emerging variability of plausible sets of values that this type of neuron can adopt underlying complex spiking characteristics. Also, the set of selected cerebellar GrC models captured spiking dynamics closer to the reference model than the single model obtained with off-the-shelf parameter optimization algorithms used in our previous article. The method hereby proposed represents a valuable strategy for adjusting a varied population of realistic and simplified neuron models. It can be applied to other kinds of neuron models and biological contexts.

Optimization of Efficient Neuron Models With Realistic Firing Dynamics. The Case of the Cerebellar Granule Cell.

Biologically relevant large-scale computational models currently represent one of the main methods in neuroscience for studying information processing primitives of brain areas. However, biologically realistic neuron models tend to be computationally heavy and thus prevent these models from being part of brain-area models including thousands or even millions of neurons. The cerebellar input layer represents a canonical example of large scale networks. In particular, the cerebellar granule cells, the most numerous cells in the whole mammalian brain, have been proposed as playing a pivotal role in the creation of somato-sensorial information representations. Enhanced burst frequency (spiking resonance) in the granule cells has been proposed as facilitating the input signal transmission at the theta-frequency band (4-12 Hz), but the functional role of this cell feature in the operation of the granular layer remains largely unclear. This study aims to develop a methodological pipeline for creating neuron models that maintain biological realism and computational efficiency whilst capturing essential aspects of single-neuron processing. Therefore, we selected a light computational neuron model template (the adaptive-exponential integrate-and-fire model), whose parameters were progressively refined using an automatic parameter tuning with evolutionary algorithms (EAs). The resulting point-neuron models are suitable for reproducing the main firing properties of a realistic granule cell from electrophysiological measurements, including the spiking resonance at the theta-frequency band, repetitive firing according to a specified intensity-frequency (I-F) curve and delayed firing under current-pulse stimulation. Interestingly, the proposed model also reproduced some other emergent properties (namely, silent at rest, rheobase and negligible adaptation under depolarizing currents) even though these properties were not set in the EA as a target in the fitness function (FF), proving that these features are compatible even in computationally simple models. The proposed methodology represents a valuable tool for adjusting AdEx models according to a FF defined in the spiking regime and based on biological data. These models are appropriate for future research of the functional implication of bursting resonance at the theta band in large-scale granular layer network models.

An integrative methodology based on protein-protein interaction networks for identification and functional annotation of disease-relevant genes applied to channelopathies.

BACKGROUND: Biologically data-driven networks have become powerful analytical tools that handle massive, heterogeneous datasets generated from biomedical fields. Protein-protein interaction networks can identify the most relevant structures directly tied to biological functions. Functional enrichments can then be performed based on these structural aspects of gene relationships for the study of channelopathies. Channelopathies refer to a complex group of disorders resulting from dysfunctional ion channels with distinct polygenic manifestations. This study presents a semi-automatic workflow using protein-protein interaction networks that can identify the most relevant genes and their biological processes and pathways in channelopathies to better understand their etiopathogenesis. In addition, the clinical manifestations that are strongly associated with these genes are also identified as the most characteristic in this complex group of diseases. RESULTS: In particular, a set of nine representative disease-related genes was detected, these being the most significant genes in relation to their roles in channelopathies. In this way we attested the implication of some voltage-gated sodium (SCN1A, SCN2A, SCN4A, SCN4B, SCN5A, SCN9A) and potassium (KCNQ2, KCNH2) channels in cardiovascular diseases, epilepsies, febrile seizures, headache disorders, neuromuscular, neurodegenerative diseases or neurobehavioral manifestations. We also revealed the role of Ankyrin-G (ANK3) in the neurodegenerative and neurobehavioral disorders as well as the implication of these genes in other systems, such as the immunological or endocrine systems. CONCLUSIONS: This research provides a systems biology approach to extract information from interaction networks of gene expression. We show how large-scale computational integration of heterogeneous datasets, PPI network analyses, functional databases and published literature may support the detection and assessment of possible potential therapeutic targets in the disease. Applying our workflow makes it feasible to spot the most relevant genes and unknown relationships in channelopathies and shows its potential as a first-step approach to identify both genes and functional interactions in clinical-knowledge scenarios of target diseases. METHODS: An initial gene pool is previously defined by searching general databases under a specific semantic framework. From the resulting interaction network, a subset of genes are identified as the most relevant through the workflow that includes centrality measures and other filtering and enrichment databases.

Detección de Betalactamasas de espectro extendido en Enterobacteriaceae en un Centro de Salud de Maracaibo, Venezuela / Detection of Extended-Spectrum ß-lactamase-producing Enterobacteriaceae in a Healthcare Center from Maracaibo city, Venezuela

La alta incidencia de las enfermedades infecciosas y el aumento de la resistencia a los antibióticos se han convertido en la actualidad en un problema de salud pública, siendo las enterobacterias productoras de Betalactamasas de Espectro Extendido (BLEE) un ejemplo de este fenómeno. En el presente estudio se determinó la producción de BLEE en aislados clínicos de la familia Enterobacteriaceae procedentes de una institución de salud de la ciudad de Maracaibo, durante el periodo septiembre de 2014 a febrero de 2015. Para la detección de BLEE se utilizó como método preliminar el de Kirby-Baüer, siguiendo los lineamientos del CLSI; adicionalmente se utilizó como prueba confirmatoria fenotípica el método de sinergia del doble disco y como prueba confirmatoria genotípica la detección de los genes blaCTX-M, blaTEM y blaSHV mediante PCR. Se analizaron 55 enterobacterias productoras de BLEE, distribuidas de la siguiente manera: Escherichia coli 56,36%, Klebsiella pneumoniae 21,82%, Enterobacter cloacae 7,27%, Proteus mirabilis y Serratia marcescens 5,45% para cada especie, por último, Salmonella spp. y Morganella morganii 1,82% respectivamente. En cuanto al tipo de BLEE detectado mediante PCR, se observó que el 83,63% de los aislados presentó el tipo TEM, seguido de CTX-M (23,63%) y SHV (21,81%), mientras que el 27,27% de los aislados produjo dos o tres BLEE de manera simultánea. Los resultados de este estudio confirman la alta diseminación de este mecanismo de resistencia entre las enterobacterias productoras de infecciones en nuestras instituciones públicas de salud, por lo que deben aplicarse medidas de control que permitan controlar y disminuir su incidencia.

The high incidence of the infectious diseases and the antimicrobial resistance arise represent a public health threat today. The extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae are an example of this phenomenon. We determined the ESBL-production in Enterobacteriaceae isolates from a Healthcare Center in Maracaibo, during September 2014 to February 2015. The Kirby-Baüer method was perform to preliminary phenotypic detection of ESBL, according to CLSI guidelines. ESBL-production was confirmed by a double-disk synergy test according to the CLSI standards. To genotypic confirmation, the genes blaCTX-M, blaTEM and blaSHV were amplified by PCR. Fifty-five (n=55) strains were analyzed distributed in Escherichia coli (56.36 %), Klebsiella pneumoniae (21.82 %), Enterobacter cloacae (7.27 %), Proteus mirabilis and Serratia marcescens (5.45 % each one), Salmonella spp. and Morganella morganii (1.82 % each one). The major encoded ESBL was the blaTEM gene (83.63 %); followed by 23.63% of the blaCTX-M gene, and 21.81 % encoded the blaSHV gene. 27.27 % of the isolates produced two or three ESBL simultaneously. These results confirmed the high spread of this resistant mechanism among Enterobacteriaceae-producing infections in our public health institutions, therefore control measures should applied to control and reduce its incidence.

Staphylococcus aureus con resistencia múltiple a los antibióticos (MDR) en un Hospital de Maracaibo- Venezuela / Multidrug-resistant (MDR) Staphylococcus aureus in a Maracaibo’s hospital, Venezuela

Staphylococcus aureus resistente a oxacilina (SAOR) continúa siendo una causa importante de infecciones nosocomiales en todo el mundo. Se determinó la resistencia a los antibióticos de cepas intrahospitalarias, clasificándolas en multidrogo-resistentes, extensamente drogo-resistentes o pandrogo-resistentes. Las muestras biológicas fueron recolectadas entre septiembre 2013-febrero 2014 y procesadas de acuerdo a técnicas de bacteriología convencional. La resistencia a los antibióticos se determinó mediante el método de difusión con discos en agar y el gen mecA se detectó mediante reacción en cadena de la polimerasa. Se observó baja prevalencia de SAOR intrahospitalario (13,86%). La mayor resistencia fue a eritromicina (66,07%), mientras que la resistencia frente a aminoglucósidos, fluoroquinolonas, clindamicina y tetraciclina fue inferior al 25%; la resistencia frente a trimetoprim/sulfametoxazol fue muy baja y el 100% de las cepas mostraron sensibilidad a rifampicina, linezolid, vancomicina y teicoplanina. El fenotipo de resistencia a MLSB más frecuente fue el de resistencia a eritromicina y susceptibilidad a clindamicina (33,93%, fenotipo MSB). Las cepas SAOR aisladas presentaron 25 antibiotipos diferentes, siendo la mayoría de los aislamientos multidrogo-resistentes (55,36%). No se observó resistencia extensa a los antibióticos ni pandrogo-resistencia y la presencia del gen mecA se demostró en todos los aislamientos resistentes a oxacilina.

Oxacillin-resistant Staphylococcus aureus (ORSA) has remained a major cause of nosocomial infections worldwide. The antibiotic resistance of isolations was determined and we classify them in multidrug-resistant, extensively drug-resistant or pandrug-resistant. The biological samples of patients from a Maracaibo’s Hospital, during September 2013 to February 2014, were processed according to conventional techniques of bacteriology. Antibiotic resistance was determined by disk diffusion method in agar and the mecA gene was detected by polymerase chain reaction. It was observed a low prevalence of nosocomial ORSA (13.86%). The higher antibiotic resistance was observed against erythromycin (66.07%) and a resistance lower than 25% to aminoglycosides, fluoroquinolones, tetracycline and clindamycin. The isolates showed a very low resistance to trimethoprim/sulfamethoxazole and all isolates were susceptible to rifampicin, linezolid, vancomycin and teicoplanin.The majority of isolates had a MSB phenotype (33.93%), with erythromycin resistance and susceptibility to clindamycin. The ORSA isolates in this study had 25 different antibiotypes and the majority of them were multidrug-resistant (55.36%). There was not both extensively drug-resistant and pandrug-resistant isolates and the presence of the mecA gene was demonstrated in all isolates of ORSA.

Epidemiology of musculoskeletal pain in a pediatric emergency department.

The objectives of this study were (1) to determine the percentage of emergency department (ED) visits due to musculoskeletal pain (MSP) by children 3-14 years of age during a period of 1 year; (2) to determine the most frequent presenting complaints; and (3) to characterize their etiology. A cross-sectional study was performed on children aged 3-14(11/12) years attended at the ED of a tertiary hospital due to MSP. The demographic and clinical characteristics of the patients were reviewed 5 days each month for 12 consecutive months. Study days were selected by computer-generated simple random sampling. Out of 4,531 visits to the ED, 826 were due to MSP (18.2 %; 95 % CI 17.1-19.4 %). When compared with children with no skeletal complaints, children with MSP had a similar sex distribution but were older (mean ± SD 7 ± 3.5 years vs 9.9 ± 3.1 years; p < 0.0001). The most common complaints were pain at the wrist (19 %), ankle (19 %) and finger (15 %). The most common etiology was trauma (88.4 %), including contusions (38 %), fractures (21 %) and sprains (18 %). Children with hip (6.7 ± 3 years; p < 0.0001) and elbow (7.8 ± 3.5 years; p < 0.0001) complaints were younger than children with pain in other locations, whereas children with wrist pain (10.5 ± 2.6 years; p < 0.002) and joint sprains (10.7 ± 2.7 years; p < 0.0001) were older. Fractures were more frequent in boys (64 vs 36 %, p = 0.008; OR 1.6; CI 1.1-2.2). Visits to the ED due to MSP increased with age. Pain at three locations represented 50 % of the presenting complaints. Trauma was the principal etiology, but fractures only represented one-fifth of the total.

Riesgo microbiológico asociado al uso de teléfonos móviles en laboratorios clínicos hospitalarios de Maracaibo-Venezuela / Microbiological hazard associated with using of mobile phones in hospital clinical laboratories from Maracaibo-Venezuela

El uso de teléfonos móviles se ha generalizado en las instituciones de salud, inclusive en áreas con riesgos microbiológicos definidos, como los laboratorios clínicos. En el presente estudio, se investigó la presencia de bacterias potencialmente patógenas, en los teléfonos móviles del personal de laboratorio de cuatro instituciones hospitalarias de Maracaibo, Venezuela. Se realizó el cultivo microbiológico cualitativo de hisopados de la superficie de 200 teléfonos. Mediante un cuestionario se indagó la adherencia del personal a las prácticas higiénicas estándar durante la jornada laboral. En 83% de los teléfonos se evidenció contaminación bacteriana y en 29% se identificó agentes con potencial patogénico definido, predominando Enterococcus spp., anaerobios estrictos, Staphylococcus aureus y enterobacterias. Algunas cepas patógenas presentaron patrones de resistencia sugestivos de gérmenes nosocomiales. Un elevado porcentaje del personal abordado admitió no aplicar medidas higiénicas mínimas al utilizar sus teléfonos en el ambiente laboral. Los resultados microbiológicos obtenidos, aunados al bajo nivel de compromiso del personal de laboratorio con las prácticas higiénicas estándar, permiten atribuirle un importante riego microbiológico al uso de teléfonos móviles en los laboratorios clínicos, que podría afectar no sólo a los dueños y manipuladores habituales de tales dispositivos, sino también extenderse a sus manipuladores ocasionales a nivel extrahospitalario.

The use of mobile phones is widespread in health institutions, including areas with defined microbiological risk as clinical laboratories. In this study, the presence of potentially pathogenic bacteria was investigated in mobile phones of laboratory personnel of four hospitals of Maracaibo, Venezuela. Qualitative microbiological culture of swabs from the surface of 200 mobile phones was performed. A questionnaire was applied, to evaluate adherence of staff to standard hygiene practices during the workday. In 83% of phones bacterial contamination was evidenced, and 29% had bacteria with defined pathogenic potential, predominantly Enterococcus spp., strict anaerobes bacteria, Staphylococcus aureus and enterobacteria. Some pathogenic strains showed resistance patterns suggestive of nosocomial bacteria. A high percentage of staff refused to apply minimal hygiene measures to manipulate their phones during the workday. Microbiological results, analyzed together with the low level of adherence of personnel to the standard hygienic practices, allow attributing an important hazard to mobiles phone use in clinical laboratories, which could involve not only owners of such devices and usual manipulators, but also could extend to his occasional manipulators in the community.

Use of combined transmucosal fentanyl, nitrous oxide, and hematoma block for fracture reduction in a pediatric emergency department.

OBJECTIVE: The objective of this study was to evaluate the use of combined inhaled nitrous oxide (NO), hematoma block (HB), and transmucosal fentanyl (TMF) as sedoanalgesia in the reduction of radioulnar fractures in children in a pediatric emergency department (PED). METHODS: A retrospective, analytical observational study examining the cases of radioulnar fracture reduction in PED from 2007 to 2009 in children from 4 to 15 years old. The cases were divided into 2 groups: those in which only NO + HB was used and those in which TMF was combined with NO + HB. The pain perceived by the child, the doctor, and the nurse was studied during the procedure with 0- to 10-point scales (10 being severe pain). Satisfaction of the medical professionals, duration of the procedure, and the adverse effects that appeared were also studied. RESULTS: Eighty-one children were included. Sixty-four children (79%) received NO + TMF + HB, and 17 children (21%) received NO + HB only. The pain perceived by the child during the procedure in the group receiving NO + TMF + HB was 2.5 (95% confidence interval [CI], 1.8-3.1) compared with 3.9 (95% CI, 2.3-5.5) in the NO + HB group (P = 0.035), the pain perceived by the doctor was 2.6 (95% CI, 2-3.2) compared with 4 (95% CI, 1.6-4), and by the nurse was 2.7 (95% CI, 2-3.3) compared with 3.9 (95% CI, 2.3-5.5), respectively. Adverse events appeared in 15.3% of the NO + TMF + HB group and in 40% of the NO + HB group. CONCLUSIONS: The association of NO + TMF + HB in the reduction of radioulnar fractures in PED improves pain control compared with the NO + HB combination. New studies are required to confirm the benefit and safety of this drug combination.

Utilidad de las pruebas rápidas de detección antigénica en el diagnóstico de la nueva gripe A (H1N1) en niños / Rapid influenza diagnostic tests for detection of novel influenza A (H1N1) virus in children

Introducción Utilidad de la prueba rápida BD Directigen® EZ Flu A+B frente a gripe A (H1N1) en niños. Metódos Estudio descriptivo retrospectivo con 140 niños a los que se realizó la prueba rápida y la reacción en cadena de la polimerasa en tiempo real frente a gripe A (H1N1).Resultados Sensibilidad del 70,4% (IC: 58,9–79,7), especificidad de 100% (IC: 94,7–100), valor predictivo negativo del 76,6% (IC: 66,9–84,2) y valor predictivo positivo del 100% (IC: 92,8–100).Conclusión La prueba presenta buena sensibilidad y buena especificidad en niños, siendo similares a los de la gripe estacional (AU)

Introduction Accuracy of rapid BD Directigen® EZ Flu A+B diagnostic test against Influenza A (H1N1) Virus in children. Methods A descriptive retrospective study was performed. One hundred and forty children underwent the rapid influenza test and the RT-PCR against Influenza A (H1N1) Virus. Results The sensitivity was 70.4% (95% CI: 58.9–79.7), specificity 100 % (95% CI: 94.7–100), NPV 76.6% (95% CI: 66.9–84,2) and PPV 100% (95% CI: 92.8–100).Conclusion This test showed high sensitivity and specificity, very similar to the seasonal influenza, in children (AU)

[Rapid influenza diagnostic tests for detection of novel influenza A (H1N1) virus in children]. / Utilidad de las pruebas rápidas de detección antigénica en el diagnóstico de la nueva gripe A (H1N1) en niños.

INTRODUCTION: Accuracy of rapid BD Directigen(®) EZ Flu A+B diagnostic test against Influenza A (H1N1) Virus in children. METHODS: A descriptive retrospective study was performed. One hundred and forty children underwent the rapid influenza test and the RT-PCR against Influenza A (H1N1) Virus. RESULTS: The sensitivity was 70.4% (95% CI: 58.9-79.7), specificity 100 % (95% CI: 94.7-100), NPV 76.6% (95% CI: 66.9-84,2) and PPV 100% (95% CI: 92.8-100). CONCLUSION: This test showed high sensitivity and specificity, very similar to the seasonal influenza, in children.