Analysis of the early response to spinal cord injury identified a key role for mTORC1 signaling in the activation of neural stem progenitor cells.

Xenopus laevis are able to regenerate the spinal cord during larvae stages through the activation of neural stem progenitor cells (NSPCs). Here we use high-resolution expression profiling to characterize the early transcriptome changes induced after spinal cord injury, aiming to identify the signals that trigger NSPC proliferation. The analysis delineates a pathway that starts with a rapid and transitory activation of immediate early genes, followed by migration processes and immune response genes, the pervasive increase of NSPC-specific ribosome biogenesis factors, and genes involved in stem cell proliferation. Western blot and immunofluorescence analysis showed that mTORC1 is rapidly and transiently activated after SCI, and its pharmacological inhibition impairs spinal cord regeneration and proliferation of NSPC through the downregulation of genes involved in the G1/S transition of cell cycle, with a strong effect on PCNA. We propose that the mTOR signaling pathway is a key player in the activation of NPSCs during the early steps of spinal cord regeneration.

Xenopus, a Model to Study Wound Healing and Regeneration: Experimental Approaches.

Xenopus has been widely used as a model organism to study wound healing and regeneration. During early development and at tadpole stages, Xenopus is a quick healer and is able to regenerate multiple complex organs-abilities that decrease with the progression of metamorphosis. This unique capacity leads us to question which mechanisms allow and direct regeneration at stages before the beginning of metamorphosis and which ones are responsible for the loss of regenerative capacities during later stages. Xenopus is an ideal model to study regeneration and has contributed to the understanding of morphological, cellular, and molecular mechanisms involved in these processes. Nevertheless, there is still much to learn. Here we provide an overview on using Xenopus as a model organism to study regeneration and introduce protocols that can be used for studying wound healing and regeneration at multiple levels, thus enhancing our understanding of these phenomena.

Quality of the Triage of Children With Anaphylaxis at the Emergency Department.

OBJECTIVE: Early recognition of symptoms is essential in anaphylaxis management. The Canadian Paediatric Emergency Triage and Acuity Scale prioritizes anaphylaxis to level I or II (resuscitation or emergency). We analyzed the accuracy of pediatric anaphylaxis triage. METHODS: This was a retrospective review of the triage charts (adaptation of the Canadian Paediatric Emergency Triage and Acuity Scale) of 137 children attended for anaphylaxis at our pediatric emergency department. Per triages' accuracy, charts were divided into TR1 (levels I-II) and TR2 (levels III-V), comparing demographics, initial triage level given by initial assessment (Paediatric Assessment Triangle), vital signs, observations recorded by the staff, and waiting times for physician. RESULTS: Forty-six (33.3%) were triaged correctly (TR1 group), and 91 (66.7%) were not. Median ages were similar (TR1: 5 years [interquartile range, 13.1 years] vs TR2: 4.5 years [interquartile range, 14.5 years]; P = 0.837). Initial triage level 5 was given by Paediatric Assessment Triangle to 69.5% of TR1 and 83% of TR2 cases (P = 0.001; likelihood ratio for TR2: 1.985 [95% confidence interval, 1.11-3.49]). Vital signs were normal in 71.7% of TR1 and 94.5% of TR2 patients (P < 0.001; likelihood ratio for TR2: 2.602 [95% confidence interval, 1.22-5.52]). Symptoms suggestive of anaphylaxis (mention of 2 different organs) were recorded in 45.6% of TR1 and 48.3% of TR2 charts (P = 0.08). Median waiting times were 3 minutes (interquartile range, 26 minutes) and 11 minutes (interquartile range, 111 minutes) for TR1 and TR2, respectively (P = 0.001). CONCLUSIONS: Current triage, based on severity perception, missed most of the cases. Anaphylaxis-defining symptoms were overlooked. Inaccurate triage delayed medical attention. Improving measures, such as emphasizing symptom recognition and defining anaphylaxis risk discriminators, is mandatory to improve their identification.

Characterization of spinal cord damage based on automatic video analysis of froglet swimming.

Xenopus laevis frogs are a widely used organism to study aspects of modern biology ( Harland and Grainger, 2011). Its central nervous system is particularly interesting, because in certain stages of metamorphosis the spinal cord can regenerate after injury and recover swimming. With this in mind, automatic gait analysis could help evaluate the regenerative performance by means of a method that automatically and quantitatively establishes the degree in froglets' limb movement. Here, we present an algorithm that characterizes spinal cord damage in froglets. The proposed method tracks the position of the limbs throughout videos and extracts kinematic features, which posteriorly serve to differentiate froglets with different levels of damage to the spinal cord. The detection algorithm and kinematic features chosen were validated in a pattern recognition experiment in which 90 videos (divided equally in three classes: uninjured, hemisected and transected) were classified. We conclude that our system is effective in the characterization of damage to the spinal cord through video analysis of a swimming froglet with a 97% accuracy. These results potentially validate this methodology to automatically compare the recovery of spinal cord function after different treatments without the need to manually process videos. In addition, the procedure could be used to measure the kinematics and behavioral response of froglets to different experimental conditions such as nutritional state, stress, genetic background and age.

In situ injectable hydrogels for spinal cord regeneration: advances from the last 10 years.

Spinal cord injury (SCI) is a tremendously devastating disorder with no effective therapy. Neuroprotective strategies have been applied aiming to prevent secondary cell death but no successful and robust effects have been observed. Recently, combinatorial approaches using biomaterials with cells and/or growth factors have demonstrated promising therapeutic effects because of the improvement of axonal growth and in vivo functional recovery in model organisms. In situ injectable hydrogels are a particularly attractive neuroregenerative approach to improve spinal cord repair and regeneration since they can be precisely injected into the lesion site filling the space prior to gelification, decrease scarring and promote axon growth due to the hydrogel's soft structure. Important advances regarding the use of hydrogels as potential therapeutic approaches has been reported during the last 10 years. Injectable alginate hydrogel loaded with GDNF, thermoresponsives heparin-poloxamer loaded with NGF and imidazole-poly(organophosphazenes) hydrogels are just three examples of biomaterials that can promote neurite, axon growth and improve functional recovery in hemisected and resected rats. Here we will review the status of in situ injectable hydrogels for spinal cord regeneration with special focus in the advantages of using hydrogel scaffolds, the ideal polymers to be used, the gelification process and the cells or growth factors combined. The in vitro and in vivo results reported for those biomaterials will be presented, compared and discussed.

Antitumor activity and carrier properties of novel hemocyanins coupled to a mimotope of GD2 ganglioside.

Conjugation to carrier proteins is a way to improve the immunogenicity of peptides. Such is the case for peptides mimicking carbohydrate tumor-associated antigens in cancer vaccine development. The most used protein for this purpose is the keyhole limpet hemocyanin (KLH) from Megathura crenulata. Its limited bioavailability has prompted interest in finding new candidates; nevertheless, it is not known whether other hemocyanins might be equally efficient as carrier of carbohydrate peptide mimotopes to promotes anti-tumor responses. Here, we evaluated the carrier and antitumor activity of novel hemocyanins with documented immunogenicity obtained from Concholepas concholepas (CCH) and Fissurella latimarginata (FLH), coupled through sulfo-SMCC to P10, a mimetic peptide of GD2, the major ganglioside constituent of neuroectodermal tumors, and incorporating AddaVax as an adjuvant. The humoral immune responses of mice showed that CCH-P10 and FLH-P10 conjugates elicited specific IgM and IgG antibodies against P10 mimotope, similar to those obtained with KLH-P10, which was used as a positive control. The CCH-P10 and FLH-P10 antisera, exhibited cross-reactivity with murine and human melanoma cells, like anti-CCH and anti-FLH sera suggesting a cross-reaction of CCH and FLH glycosylations with carbohydrate epitopes on the tumor cell surfaces, similar to the KLH antisera. When mice were primed with each hemocyanin-P10 and challenged with melanoma cells, better antitumor effects were observed for FLH-P10 than for CCH-P10 and, as for KLH-P10, irrespective of conjugation. These data demonstrate that CCH and FLH are useful carriers of carbohydrate mimotopes; however, the best antitumor activity of FLH preparations, indicate that is a suitable candidate for further cancer vaccines research.

Impact of a working group on gastrointestinal decontamination in Spanish emergency departments.

INTRODUCTION AND OBJECTIVES: The aim of this study is to analyze the impact of the actions carried out by the Intoxications Working Group (IWG) of Spanish Society of Pediatric Emergencies in the management of acute pediatric intoxications in Spain, specifically the publishing of a Handbook in 2004 or the creation of the Toxicologic Surveillance System in 2009. PATIENTS AND METHODS: Gastrointestinal decontamination procedures were analyzed in three periods of time in Pediatric Emergency Departments (PEDs) included in the IWG: group A (2001-2002, 17 PED, 2157 episodes), group B (2008-2009, 22 PED, 612 episodes), and group C (2009-2011, 42 PED, 400 episodes). These periods were chosen because the main actions of the IWG were developed in the time in-between them. RESULTS: Of the 3169 episodes included, a gastrointestinal decontamination procedure was performed in 1031. The use of ipecac syrup decreased from 22.8% in group A to 0 in group C and the performance of a gastric lavage decreased from 29.1% in group A to 26% in group C (NS), although on splitting yearly patients of group C, it decreased to 14.7% in 2011. CONCLUSION: Recommendations developed and spread by a Working Group have approached the management of acute pediatric intoxications in Spain to international guidelines on the basis of scientific evidence.

Accuracy of a sequential approach to identify young febrile infants at low risk for invasive bacterial infection.

INTRODUCTION: Much effort has been put in the past years to create and assess accurate tools for the management of febrile infants. However, no optimal strategy has been so far identified. A sequential approach evaluating, first, the appearance of the infant, second, the age and result of the urinanalysis and, finally, the results of the blood biomarkers, including procalcitonin, may better identify low risk febrile infants suitable for outpatient management. OBJECTIVE: To assess the value of a sequential approach ('step by step') to febrile young infants in order to identify patients at a low risk for invasive bacterial infections (IBI) who are suitable for outpatient management and compare it with other previously described strategies such as the Rochester criteria and the Lab-score. METHODS: A retrospective comparison of three different approaches (step by step, Lab-score and Rochester criteria) was carried out in 1123 febrile infants less than 3 months of age attended in seven European paediatric emergency departments. IBI was defined as isolation of a bacterial pathogen from the blood or cerebrospinal fluid. RESULTS: Of the 1123 infants (IBI 48; 4.2%), 488 (43.4%) were classified as low-risk criteria according to the step by step approach (vs 693 (61.7%) with the Lab-score and 458 (40.7%) with the Rochester criteria). The prevalence of IBI in the low-risk criteria patients was 0.2% (95% CI 0% to 0.6%) using the step by step approach; 0.7% (95% CI 0.1% to 1.3%) using the Lab-score; and 1.1% (95% CI 0.1% to 2%) using the Rochester criteria. Using the step by step approach, one patient with IBI was not correctly classified (2.0%, 95% CI 0% to 6.12%) versus five using the Lab-score or Rochester criteria (10.4%, 95% CI 1.76% to 19.04%). CONCLUSIONS: A sequential approach to young febrile infants based on clinical and laboratory parameters, including procalcitonin, identifies better patients more suitable for outpatient management.

Diagnostic value of procalcitonin in well-appearing young febrile infants.

BACKGROUND AND OBJECTIVE: Procalcitonin (PCT) has been introduced in many European protocols for the management of febrile children. Its value among young, well-appearing infants, however, is not completely defined. Our objective was to assess its performance in diagnosing serious bacterial infections and specifically invasive bacterial infections (IBIs) in well-appearing infants aged <3 months with fever without source (FWS). METHODS: Well-appearing infants aged <3 months with FWS admitted to 7 European pediatric emergency departments were retrospectively included. IBI was defined as the isolation of a bacterial pathogen in blood or cerebrospinal fluid culture. RESULTS: We included 1112 infants who had PCT measured and a blood culture performed. IBI was diagnosed in 23 cases (2.1%). In the multivariate analysis including clinical and laboratory data, PCT was the only independent risk factor for IBI (odds ratio 21.69; 95% confidence interval [CI] 7.93-59.28 for PCT ≥ 0.5 ng/mL). Positive likelihood ratios for PCT ≥ 2 ng/mL and C-reactive protein (CRP) >40 mg/L were 11.14 (95% CI 7.81-15.89) and 3.45 (95% CI 2.20-5.42), respectively. Negative likelihood ratios for PCT <0.5 ng/mL and CRP <20 mg/L were 0.25 (95% CI 0.12-0.55) and 0.41 (95% CI 0.22-0.76). Among patients with normal urine dipstick results and fever of recent onset, areas under the receiver operator characteristic curve for PCT and CRP were 0.819 and 0.563, respectively. CONCLUSIONS: Among well-appearing young infants with FWS, PCT performs better than CRP in identifying patients with IBIs and seems to be the best marker for ruling out IBIs. Among patients with normal urine dipstick results and fever of recent onset, PCT remains the most accurate blood test.

Diagnostic performance of the lab-score in predicting severe and invasive bacterial infections in well-appearing young febrile infants.

BACKGROUND AND OBJECTIVES: The "Lab-score" combining C-reactive protein, procalcitonin and urine dipstick results has recently been derived and validated as an accurate tool for predicting severe bacterial infections (SBIs) in children with fever without source. We aimed to assess the Lab-score usefulness in predicting SBI, especially invasive bacterial infections (IBIs), in well-appearing infants <3 months with fever without source. METHODS: A multicenter retrospective study was conducted in 7 pediatric emergency departments in Spain and Italy. An SBI was defined as isolation of a bacterial pathogen from urine, blood, cerebrospinal fluid or stools, an IBI as isolation of a bacterial pathogen from blood or cerebrospinal fluid. The diagnostic characteristics of the Lab-score for detection of SBI and IBI were calculated. RESULTS: An SBI was diagnosed in 287 (28.3%) of 1012 patients and an IBI in 23 (2.1%) of 1098. The positive and negative likelihood ratios of a score ≥3 for SBI prediction were 10.2 (95% confidence interval [CI]: 9.5-10.9) and 0.5 (95% CI: 0.5-0.5), respectively. The area under the receiver operating characteristic curve was 0.83 (95% CI: 0.80-0.86). The same diagnostic accuracy measures for identification of IBI were 4.3 (95% CI: 4-4.6), 0.4 (95% CI: 0.3-0.5) and 0.85 (95% CI: 0.76-0.94), respectively. Use of Lab-score would have resulted in misdiagnosis of 7 (30%) infants with IBI. CONCLUSIONS: In well-appearing infants with fever without source, the Lab-score seems a more useful tool for ruling in, rather than ruling out, SBI. Its accuracy for IBI prediction was unsatisfactory.

Estudian las características histopatológicas y moleculares de las células de Langerhans en la mucosa bucal / Langerhans cells in oral mucosa: hystopatological and molecular characterization

Actualmente, las células dendríticas en tejidos periféricos, piel y mucosas son centro de numerosas publicaciones. De acuerdo con su localización se clasifican en subtipos de diferente denominación. Su posición apropiada les permite ejercer un papel crucial en la detección y la captación de antígenos y su presentación a los linfocitos T en centros linfoides, iniciando la inmunidad innata y la inmunidad adaptativa. Debe destacarse además su actuación en la tolerancia inmunitaria y su participación en diversas enfermedades autoinmunitarias como la enfermedad periodontal e incluso el cáncer de células escamosas. Presentamos una actualización resumida de los últimos estudios realizados y destacamos el conocimiento de los marcadores inmunológicos y las características fundamentales de los subtipos, células de Langerhans y células plasmocitoides, especialmente en la mucosa oral. La disminución de los distintos fenotipos de células dendríticas en lesiones cancerizables y en la vejez, como su ausencia en neoplasias malignas son aspectos destacables. Comunicamos a la vez nuestro primer acercamiento a la microscopia electrónica de barrido para observarlas en tejido mucoso sano y de tres lesiones: mucosa hiperplásica, gingivitis descamativa y úlceras aftosas recidivantes con vasculitis.

Evaluación de disfunción tiroidea según valores de TSH en una población del Centro de Atención Ambulatorio (CAA) Central Quito del Instituto Ecuatoriano de Seguridad Social, durante el año 2008 / Evaluation of thyroid dysfunction according to TSH values ​​in a population of the Center of Ambulatory Attention (CAA) Central Quito del Ecuadorian Social Security Institute, during 2008

Introducción: La cuantificación de la hormona estimulante de la tiroides (TSH) es una prueba confiable para detectar patologías tiroideas. En los pacientes que acuden por otras causas a nuestra unidad no existen datos epidemiológicos publicados. Objetivo: Determinar la frecuencia de disfunción tiroidea en la población que acude al Centro de Atención Ambulatorio (CAA) Central del Instituto Ecuatoriano de Seguridad Social (IESS). Métodos: Estudio retrospectivo que incluyó 270 mujeres y 96 hombres adultos, sin antecedentes de patología tiroidea ni consumo de hormona tiroidea, referidos por orden médica al laboratorio clínico del CAA Central Quito del IESS para la determinación por primera vez de niveles séricos de TSH por electroquimioluminiscencia. Resultados: La edad promedio de la población general fue de 55.97 (DE 14.80) años, sin diferencias significativa entre hombre y mujeres. El valor de TSH promedio para toda la muestra fue 3.08 mUI/L (DE 8.8; rango 0.01 a 32.77), sin diferencias significativas según sexo. Se encontró valores de TSH <0.27 mUI/L en 8 pacientes (2.18%), en rango normal (0.27-4.2 mUI/L) en 303 pacientes (82.78%), > 4.2 mUI/L pero < 10 mUI/L en 49 pacientes (13.38%) y niveles mayores de 10 mUI/L en 6 pacientes (1.63%). Se hallaron niveles elevados de TSH con mayor frecuencia a partir de la cuarta década de vida. Conclusión: Se justifica una búsqueda más activa de hipotiroidismo y su realización como prueba de tamizaje en mujeres mayores de 40 años. Los valores hallados de disfunción tiroidea solo aplican para esta población evaluada, en la cual se utilizó más una estrategia diagnóstica que de tamizaje y cuyas incidencias no pueden extrapolarse a la población general.

Introduction: The quantification of thyroid stimulating hormone (TSH) is a reliable test to detect thyroid pathologies. In the patients who come For other reasons, our unit does not have published epidemiological data. Objective: To determine the frequency of thyroid dysfunction in the population that attends to the Central Ambulatory Care Center (CAA) of the Ecuadorian Institute of Social Security (IESS). Methods: Retrospective study that included 270 women and 96 adult men, without history of thyroid pathology or thyroid hormone consumption, referred in order to the clinical laboratory of the CAA Central Quito of the IESS for the determination for the first time of serum TSH levels by electrochemiluminescence. Results: The average age of the general population was 55.97 (SD 14.80) years, without significant differences between men and women. The average TSH value for the entire sample it was 3.08 mUI / L (DE 8.8; range 0.01 to 32.77), without differences significant according to sex. TSH values ​​<0.27 mUI / L were found in 8 patients (2.18%), in normal range (0.27-4.2 mUI / L) in 303 patients (82.78%),> 4.2 mUI / L but <10 mUI / L in 49 patients (13.38%) and levels greater than 10 mUI / L in 6 patients (1.63%). Elevated levels of TSH were found more frequently at from the fourth decade of life. Conclusion: A more active search for hypothyroidism and its realization is justified as a screening test in women over 40 years old. The values ​​found of Thyroid dysfunction only applies to this population evaluated, in which more was used a diagnostic strategy that screening and whose incidents cannot be extrapolated to the general population.