Small extracellular vesicle targeting of hypothalamic AMPKα1 promotes weight loss in leptin receptor deficient mice.

BACKGROUND AND AIMS: Leptin receptor (LEPR) deficiency promotes severe obesity and metabolic disorders. However, the current therapeutic options against this syndrome are scarce. METHODS: db/db mice and their wildtypes were systemically treated with neuronal-targeted small extracellular vesicles (sEVs) harboring a plasmid encoding a dominant negative mutant of AMP-activated protein kinase alpha 1 (AMPKα1-DN) driven by steroidogenic factor 1 (SF1) promoter; this approach allowed to modulate AMPK activity, specifically in SF1 cells of the ventromedial nucleus of the hypothalamus (VMH). Animals were metabolically phenotyped. RESULTS: db/db mice intravenously injected with SF1-AMPKα1-DN loaded sEVs showed a marked feeding-independent weight loss and decreased adiposity, associated with increased sympathetic tone, brown adipose tissue (BAT) thermogenesis and browning of white adipose tissue (WAT). CONCLUSION: Overall, this evidence indicates that specific modulation of hypothalamic AMPK using a sEV-based technology may be a suitable strategy against genetic forms of obesity, such as LEPR deficiency.

BMP8 and activated brown adipose tissue in human newborns.

The classical dogma states that brown adipose tissue (BAT) plays a major role in the regulation of temperature in neonates. However, although BAT has been studied in infants for more than a century, the knowledge about its physiological features at this stage of life is rather limited. This has been mainly due to the lack of appropriate investigation methods, ethically suitable for neonates. Here, we have applied non-invasive infrared thermography (IRT) to investigate neonatal BAT activity. Our data show that BAT temperature correlates with body temperature and that mild cold stimulus promotes BAT activation in newborns. Notably, a single short-term cold stimulus during the first day of life improves the body temperature adaption to a subsequent cold event. Finally, we identify that bone morphogenic protein 8B (BMP8B) is associated with the BAT thermogenic response in neonates. Overall, our data uncover key features of the setup of BAT thermogenesis in newborns.

A New Biomarker Tool for Risk Stratification in "de novo" Acute Heart Failure (OROME).

Background: Inflammation is one of the mechanisms involved in heart failure (HF) pathophysiology. Thus, the acute phase reactant protein, orosomucoid, was associated with a worse post-discharge prognosis in de novo acute HF (AHF). However, the presence of anti-inflammatory adipokine, omentin, might protect and reduce the severity of the disease. We wanted to evaluate the value of omentin and orosomucoid combination for stratifying the risk of these patients. Methods and Results: Two independent cohorts of patients admitted for de novo AHF in two centers were included in the study (n = 218). Orosomucoid and omentin circulating levels were determined by ELISA at discharge. Patients were followed-up for 317 (3-575) days. A predictive model was determined for the primary endpoint, death, and/or HF readmission. Differences in survival were evaluated using a Log-rank test. According to cut-off values of orosomucoid and omentin, patients were classified as UpDown (high orosomucoid and low omentin levels), equal (both proteins high or low), and DownUp (low orosomucoid and high omentin levels). The Kaplan Meier determined a worse prognosis for the UpDown group (Long-rank test p = 0.02). The predictive model that includes the combination of orosomucoid and omentin groups (OROME) + NT-proBNP values achieved a higher C-index = 0.84 than the predictive model with NT-proBNP (C-index = 0.80) or OROME (C-index = 0.79) or orosomucoid alone (C-index = 0.80). Conclusion: The orosomucoid and omentin determination stratifies de novo AHF patients into the high, mild, and low risk of rehospitalization and/or death for HF. Its combination with NT-proBNP improves its predictive value in this group of patients.

Intravitreal injections of anti-VEGF agents and antibiotic prophylaxis for endophthalmitis: A systematic review and meta-analysis.

We performed a systematic review and meta-analysis to determine whether the use of local antibiotics is a beneficial prophylactic treatment for endophthalmitis in patients treated with anti-VEGF agents. We searched the MEDLINE and EMBASE databases, and the Cochrane Library over the period January 2007 to December 2016. The search terms used included "Endophthalmitis", "Antibiotic" and "Intravitreal injection". Studies in which the patients were treated exclusively with intravitreal injections of anti-VEGF were selected. Eight studies fit the inclusion criteria, which included a total of 276,774 injections; 109,178 (39.45%) were associated with the use of antibiotics and 114,821 (60.55%) were not associated with the use of antibiotics. Our meta-analysis indicated a significant risk for endophthalmitis that was 1.70 times greater with the use of antibiotics than that without antibiotics, with a confidence interval of 1.08 to 2.66 (p = 0.02). A meta-regression indicated that the location (operating rooms versus outpatient clinics) of injection did not have a significant effect on the incidence of endophthalmitis. The prophylactic use of antibiotics when administering anti-VEGF intravitreal injections may contribute to a greater incidence of endophthalmitis. This finding, in addition to reducing costs, would eliminate a treatment that has been shown to be unnecessary and even harmful to patients.

Analysis of the diversity of substrate utilisation of soil bacteria exposed to Cd and earthworm activity using generalised additive models.

Biolog EcoPlates™ can be used to measure the carbon substrate utilisation patterns of microbial communities. This method results in a community-level physiological profile (CLPP), which yields a very large amount of data that may be difficult to interpret. In this work, we explore a combination of statistical techniques (particularly the use of generalised additive models [GAMs]) to improve the exploitation of CLPP data. The strength of GAMs lies in their ability to address highly non-linear relationships between the response and the set of explanatory variables. We studied the impact of earthworms (Aporrectodea caliginosa Savigny 1826) and cadmium (Cd) on the CLPP of soil bacteria. The results indicated that both Cd and earthworms modified the CLPP. GAMs were used to assess time-course changes in the diversity of substrate utilisation (DSU) using the Shannon-Wiener index. GAMs revealed significant differences for all treatments (compared to control -S-). The Cd exposed microbial community presented very high metabolic capacities on a few substrata, resulting in an initial acute decrease of DSU (i.e. intense utilization of a few carbon substrata). After 54 h, and over the next 43 h the increase of the DSU suggest that other taxa, less dominant, reached high numbers in the wells containing sources that are less suitable for the Cd-tolerant taxa. Earthworms were a much more determining factor in explaining time course changes in DSU than Cd. Accordingly, Ew and EwCd soils presented similar trends, regardless the presence of Cd. Moreover, both treatments presented similar number of bacteria and higher than Cd-treated soils. This experimental approach, based on the use of DSU and GAMs allowed for a global and statistically relevant interpretation of the changes in carbon source utilisation, highlighting the key role of earthworms on the protection of microbial communities against the Cd.

A novel method to handle the effect of uneven sampling effort in biodiversity databases.

How reliable are results on spatial distribution of biodiversity based on databases? Many studies have evidenced the uncertainty related to this kind of analysis due to sampling effort bias and the need for its quantification. Despite that a number of methods are available for that, little is known about their statistical limitations and discrimination capability, which could seriously constrain their use. We assess for the first time the discrimination capacity of two widely used methods and a proposed new one (FIDEGAM), all based on species accumulation curves, under different scenarios of sampling exhaustiveness using Receiver Operating Characteristic (ROC) analyses. Additionally, we examine to what extent the output of each method represents the sampling completeness in a simulated scenario where the true species richness is known. Finally, we apply FIDEGAM to a real situation and explore the spatial patterns of plant diversity in a National Park. FIDEGAM showed an excellent discrimination capability to distinguish between well and poorly sampled areas regardless of sampling exhaustiveness, whereas the other methods failed. Accordingly, FIDEGAM values were strongly correlated with the true percentage of species detected in a simulated scenario, whereas sampling completeness estimated with other methods showed no relationship due to null discrimination capability. Quantifying sampling effort is necessary to account for the uncertainty in biodiversity analyses, however, not all proposed methods are equally reliable. Our comparative analysis demonstrated that FIDEGAM was the most accurate discriminator method in all scenarios of sampling exhaustiveness, and therefore, it can be efficiently applied to most databases in order to enhance the reliability of biodiversity analyses.

Intraspecific variability in several isolates of Philasterides dicentrarchi (syn. Miamiensis avidus), a scuticociliate parasite of farmed turbot.

Research on intraspecific variation in ciliates is scarce, and in scuticociliate parasite of fish, virtually nonexistent. In this study, seven isolates obtained from turbots affected by scuticociliatosis in different parts of the Iberian Peninsula (northwest Spain and southwest Portugal) were morphologically and genetically characterized to investigate the intraspecific divergence in these amphizoic ciliates. The isolates were stained with ammoniacal silver carbonate and examined in an optical microscope; all were found to have the typical morphological characteristics described for Philasterides dicentrarchi (syn. Miamiensis avidus). Sixteen biometric characteristics of the seven isolates were used in a canonical discrimination analysis (CDA) to select a subset of those that best identified each isolate. Discriminant analysis indicated that the OPK3 width, length of the PM2, length of the buccal field, the body width, L:W ratio, the body length, the OPK1 width and the distance between OPK2 and OPK3 were the most important morphological variables for discriminating the isolates. The first three canonical functions accounted for 86% of the total variance. The scatter plots of the first two canonical variables grouped and separated the P. dicentrarchi isolates into five clusters. Flow cytometry analysis of isolates also indicated intraspecific polymorphisms among P. dicentrarchi isolates. Nuclear markers (a 349-bp and a 390-bp fragment of 18S rRNA and ß-tubulin genes) and a 398-bp of the mitochondrial cytocrome oxidase subunit I (Cox1) gene were then used to investigate the intraspecific genetic variation in P. dicentrarchi. Haplotype analysis and neighbour-joining phylogenies of nucleotide sequences of seven isolates revealed a high degree of intraspecific genetic variation among the isolates. Analysis of Cox1 and ß-tubulin genes revealed six haplotypes (and clusters) in both cases; however, analysis of the 18S rRNA gene revealed only two haplotypes. The results show clear intraspecific variation at morphological and genetic levels in the scuticociliate P. dicentrarchi, and verify the suitability of mitochondrial (Cox1) and nuclear (ß-tubulin) genes for detecting intraspecific genetic variation within populations of scuticociliates that infect cultured turbot. The existence of this intraspecific variation must be taken into account in the design of an effective vaccine to control scuticociliatosis.

Characteristics and dynamics of Salmonella contamination along the coast of Agadir, Morocco.

The occurrence of Salmonella enterica in the environment of tropical and desert regions has remained largely uninvestigated in many areas of the world, including Africa. In the present study, we investigated the presence of Salmonella spp. along 122 km of the coastline of Agadir (southern Morocco) in relation to environmental parameters. A total of 801 samples of seawater (243), marine sediment (279), and mussels (279) were collected from six sites between July 2004 and May 2008. The overall prevalence of Salmonella spp. was 7.1%, with the highest occurrence in mussels (10%), followed by sediment (6.8%) and seawater (4.1%). Only three serotypes were identified among the 57 Salmonella sp. strains isolated. S. enterica serotype Blockley represented 43.8% of all Salmonella strains and was identified in mussel and sediment samples. S. enterica serotype Kentucky (29.8%) was found almost exclusively in mussels, whereas S. enterica serotype Senftenberg (26.3%) was detected in sediment and seawater. Statistical analysis using generalized additive models identified seawater temperature, environmental temperature, rainfall, and solar radiation as significant factors associated with the presence of Salmonella. Rainfall was the only variable showing a linear positive effect on the presence of Salmonella in the sea, whereas the remaining variables showed more complex nonlinear effects. Twenty-eight (49.1%) Salmonella isolates displayed resistance to ampicillin (22 isolates), nalidixic acid (9 isolates), sulfonamide compounds (2 isolates), and tetracycline (1 isolate), with six of these isolates displaying multiple resistance to two of these antimicrobial agents. Pulsed-field gel electrophoresis analysis revealed homogenous restriction patterns within each serotype that were uncorrelated with the resistance pattern profiles.

El desarrollo e implementación de la resonancia magnética intraoperatoria en neurocirugía (remain) y su aplicación en 83 intervenciones en la Argentina / Development of REMAIN for neurosurgery and application in 83 operative intervention in Argentina

Objective: To show implementation and development of an operating room in which we operated 83 patients using intraoperative MRI (REMAIN). Method: We used a side-opening-magnet, 0.23 Tesla, installed in a surgical area specially designed with all the advances of the modern operating rooms. Results: A great variety of neuro-surgical procedures can be made with REMAIN controls. The obtained images are clear, without devices and with an excellent definition of the anatomical structures and the pathology, that allows the neurosurgeon to make more precise and safer interventions. Conclusions: The images of REMAIN in a surgical scope, make possible that injuries can be identified and located with absolute precision. It is particularly useful in determining with exactitude the tumor-like limits, optimizing the surgical approaches, obtaining complete extirpations of brain injuries and controlling the possible intraoperative complications.

Indice de Proliferación Celular (MIB-1) relacionado con el Pronóstico de los Astrocinomas Malignos en Niños y Adultos

El objetivo de este trabajo es analizar los astrocitomas de alto grado que se evaluaron en el período comprendido entre Enero de 1991 hasta Diciembre del 2001 en el Hospital Nacional Prof. Alejandro Posadas; determinar el índice de proliferación celular y asociarlo con la sobrevida de los pacientes. Material y Método: se seleccionaron 58 pacientes con gliomas cerebrales de alto grado, se estudiaron sus características clínicas y se determinó el grado de resección quirúrgica. Se clasificaron como Astrocitoma grado III o IV siguiendo los criterios de Daumas-Duport y la OMS. Se realizaron nuevos cortes y procesaron con Ac.Monoclonal K167/MIB1. Se determinó el índice de proliferación celular contando las células marcadas en varios campos de 5 y 10 aumentos (aproximadamente 1000 células) y se calculo el promedio de ellas. Se consignó el tratamiento oncológico realizado (radio y quimioterapia). Resultados: la cohorte retrospectiva está compuesta por 58 pacientes entre 2 y 76 años de edad, con una mediana de 54. El grupo adulto está integrado por 50 pacientes y el pediátrico por 8 .Con criterios de Astrocitoma Anaplásico se diagnosticaron 16 casos, compatibles con Glioblastoma Multiforme 39; gliomas mixtos en 3 casos. El índice de proliferación celular (IPC) presenta amplia dispersión con valores entre 1.0 y 40.0 por ciento, con una mediana de 6,0 por ciento para los astrocitomas de tercer grado y 14,5 por ciento para los Glioblastoma. La sobrevida (mediana) en los adultos fue de 214 días y en los niños de 1107 días. Observamos que los pacientes que presentaron IPC menor o igual a 18 por ciento presentaron 214 días de sobrevida, aquellos con IPC mayor a 18 por ciento presentaron 274 días de supervivencia (mediana). Los casos con supervivencia mas prolongadas son aquellos que tuvieron resección parcial y que posteriormente recibieron radio y quimioterapia. Cooclusiones: la clasificación histológica de los gliomas de alto grado puede ser complementada con la determinación del índice de proliferación celular(KI67/MIB.Observamos que el IPC no se asoció directamente con la sobrevida especialmente de los niños. Los casos con IPC >18 por ciento presentaron sobrevida más prolongadas. Conocer los valores de proliferación celular de un tumor puede ayudar a valorar más integralmente el futuro de los pacientes, particularmente de los que presentan valores de proliferación elevados y mal pronóstico con los tratamientos habituales. H

Indice de Proliferación Celular (MIB-1) relacionado con el Pronóstico de los Astrocinomas Malignos en Niños y Adultos

El objetivo de este trabajo es analizar los astrocitomas de alto grado que se evaluaron en el período comprendido entre Enero de 1991 hasta Diciembre del 2001 en el Hospital Nacional Prof. Alejandro Posadas; determinar el índice de proliferación celular y asociarlo con la sobrevida de los pacientes. Material y Método: se seleccionaron 58 pacientes con gliomas cerebrales de alto grado, se estudiaron sus características clínicas y se determinó el grado de resección quirúrgica. Se clasificaron como Astrocitoma grado III o IV siguiendo los criterios de Daumas-Duport y la OMS. Se realizaron nuevos cortes y procesaron con Ac.Monoclonal K167/MIB1. Se determinó el índice de proliferación celular contando las células marcadas en varios campos de 5 y 10 aumentos (aproximadamente 1000 células) y se calculo el promedio de ellas. Se consignó el tratamiento oncológico realizado (radio y quimioterapia). Resultados: la cohorte retrospectiva está compuesta por 58 pacientes entre 2 y 76 años de edad, con una mediana de 54. El grupo adulto está integrado por 50 pacientes y el pediátrico por 8 .Con criterios de Astrocitoma Anaplásico se diagnosticaron 16 casos, compatibles con Glioblastoma Multiforme 39; gliomas mixtos en 3 casos. El índice de proliferación celular (IPC) presenta amplia dispersión con valores entre 1.0 y 40.0 por ciento, con una mediana de 6,0 por ciento para los astrocitomas de tercer grado y 14,5 por ciento para los Glioblastoma. La sobrevida (mediana) en los adultos fue de 214 días y en los niños de 1107 días. Observamos que los pacientes que presentaron IPC menor o igual a 18 por ciento presentaron 214 días de sobrevida, aquellos con IPC mayor a 18 por ciento presentaron 274 días de supervivencia (mediana). Los casos con supervivencia mas prolongadas son aquellos que tuvieron resección parcial y que posteriormente recibieron radio y quimioterapia. Cooclusiones: la clasificación histológica de los gliomas de alto grado puede ser complementada con la determinación del índice de proliferación celular(KI67/MIB.Observamos que el IPC no se asoció directamente con la sobrevida especialmente de los niños. Los casos con IPC >18 por ciento presentaron sobrevida más prolongadas. Conocer los valores de proliferación celular de un tumor puede ayudar a valorar más integralmente el futuro de los pacientes, particularmente de los que presentan valores de proliferación elevados y mal pronóstico con los tratamientos habituales. H

Indice de Proliferación Celular (MIB-1) relacionado con el Pronóstico de los Astrocinomas Malignos en Niños y Adultos

El objetivo de este trabajo es analizar los astrocitomas de alto grado que se evaluaron en el período comprendido entre Enero de 1991 hasta Diciembre del 2001 en el Hospital Nacional Prof. Alejandro Posadas; determinar el índice de proliferación celular y asociarlo con la sobrevida de los pacientes. Material y Método: se seleccionaron 58 pacientes con gliomas cerebrales de alto grado, se estudiaron sus características clínicas y se determinó el grado de resección quirúrgica. Se clasificaron como Astrocitoma grado III o IV siguiendo los criterios de Daumas-Duport y la OMS. Se realizaron nuevos cortes y procesaron con Ac.Monoclonal K167/MIB1. Se determinó el índice de proliferación celular contando las células marcadas en varios campos de 5 y 10 aumentos (aproximadamente 1000 células) y se calculo el promedio de ellas. Se consignó el tratamiento oncológico realizado (radio y quimioterapia). Resultados: la cohorte retrospectiva está compuesta por 58 pacientes entre 2 y 76 años de edad, con una mediana de 54. El grupo adulto está integrado por 50 pacientes y el pediátrico por 8 .Con criterios de Astrocitoma Anaplásico se diagnosticaron 16 casos, compatibles con Glioblastoma Multiforme 39; gliomas mixtos en 3 casos. El índice de proliferación celular (IPC) presenta amplia dispersión con valores entre 1.0 y 40.0 por ciento, con una mediana de 6,0 por ciento para los astrocitomas de tercer grado y 14,5 por ciento para los Glioblastoma. La sobrevida (mediana) en los adultos fue de 214 días y en los niños de 1107 días. Observamos que los pacientes que presentaron IPC menor o igual a 18 por ciento presentaron 214 días de sobrevida, aquellos con IPC mayor a 18 por ciento presentaron 274 días de supervivencia (mediana). Los casos con supervivencia mas prolongadas son aquellos que tuvieron resección parcial y que posteriormente recibieron radio y quimioterapia. Cooclusiones: la clasificación histológica de los gliomas de alto grado puede ser complementada con la determinación del índice de proliferación celular(KI67/MIB.Observamos que el IPC no se asoció directamente con la sobrevida especialmente de los niños. Los casos con IPC >18 por ciento presentaron sobrevida más prolongadas. Conocer los valores de proliferación celular de un tumor puede ayudar a valorar más integralmente el futuro de los pacientes, particularmente de los que presentan valores de proliferación elevados y mal pronóstico con los tratamientos habituales. H