Effects of two platelet-rich gel supernatants at two concentrations on healthy cartilage explants from horses

Platelet-rich plasma is used as a treatment of arthropathies in horses. However, there is lack of in vitro information about the mechanism of action of this substance and its effects on healthy cartilage of these animals. The aims were: (1) to evaluate the effects at 48 and 96 h of two concentrations (25 and 50%) of leukocyte- and platelet-rich gel (L-PRG) and pure platelet rich gel (P-PRG) supernatants on the production/degradation in normal cartilage explants (CEs) of platelet-derived growth factor isoform BB, transforming growth factor beta-1, tumour necrosis factor alpha, interleukin (IL) 4 (IL-4), and IL-1 receptor antagonist (IL-1ra); and (2) to study possible correlations of these molecules with their respective PRG supernatant treatments. CEs from six horses were cultured for 96 h with L-PRG and P-PRG supernatants at 25 and 50% concentrations, respectively. CE culture media were changed each 48 h and used for determination, by ELISA, of the molecules. L-PRG and P-PRG supernatants at 25 and 50% concentrations influenced the molecular anti-inflammatory profile of CE groups cultured with these substances. 50% L-PRG supernatant produced the most robust pro-inflammatory effects when compared to the CE control group and the CE group treated with the other PRG supernatant concentrations. In general, PRG supernatants induced pro-inflammatory effects in normal CEs

Isolation and Identification of an Antimutagenic Phthalate Derivative Compound from Octopus (Paraoctopus limaculatus)

Purpose: To isolate and evaluate the antimutagenic properties of compounds previously identified in octopus (Paraoctopus limaculatus).Methods: Octopus fractions, previously obtained by a sequential thin layer chromatography (TLC) procedure, were subjected to further fractionation by TLC and their anti-mutagenic activity monitored using Salmonella tester strains TA98 and TA100 with metabolic activation (S9) in Ames test. The isolated fractions were subjected to structural studies by Fourier transformed infrared spectroscopy (FTIR), nuclear magnetic resonance (1H and 13C NMR), and gas chromatography-mass spectrometry.Results: Five new fractions were obtained from a previously isolated and reported anti-mutagenic octopus fraction. Fractions RB21321b2 and RB21321b3 inhibited > 80 % of the mutagenicity induced by 500 ng AFB1 on both tester strains and were selected for chemical/structural characterization. Data from IR and 1H and 13C NMR suggested the presence of phthalate type of compounds. GC-MS analysis revealed 278 m/z for both fractions which is consistent with a butyl isobutyl phthalate structure.Conclusion: Based on the findings, the compound responsible for the high anti-mutagenic activity of the isolated fraction from octopus is 1-butyl-2-isobutyl-phthalate.Keywords: Octopus, Anti-Mutagenic, Paraoctopus limaculatus, 1-Butyl-2-isobutyl-phthalat

Caracterización beta, gamma y de oscilaciones de alta frecuencia para localización de diana en procedimientos de Estimulación Cerebral Profunda

Deep Brain Stimulation (DBS) has been successfully used to treat patients with Parkinson’s Disease. DBS employs an electrode that regulates the oscillatory activity of the basal ganglia, such as the subthalamic nucleus (STN). A critical point during the surgical implantation of such electrode is the precise localization of the target. This is done using presurgical images, stereotactic frames, and microelectrode recordings (MER). The latter allows neurophysiologists to visualize the electrical activity of different structures along the surgical track, each of them with well-defined variations in the frequency pattern; however, this is far from an automatic or semi-automatic method to help these specialists make decisions concerning the surgical target. To pave the way to automation, we analyzed three frequency bands in MER signals acquired from 11 patients undergoing DBS: beta (13-40 Hz), gamma (40-200 Hz), and high-frequency oscillations (HFO – 201-400 Hz). In this study, we propose and assess five indexes in order to detect the STN: variations in autoregressive parameters and their derivative along the surgical track, the energy of each band calculated using the Yule-Walker power spectral density, the high-to-low (H/L) ratio, and its derivative. We found that the derivative of one parameter of the beta band and the H/L ratio of the HFO/gamma bands produced errors in STN targeting like those reported in the literature produced by image-based methods (<2 mm). Although the indexes introduced here are simple to compute and could be applied in real time, further studies must be conducted to be able to generalize their results.La estimulación cerebral profunda (DBS por sus siglas en inglés) ha sido usada exitosamente en el tratamiento de pacientes con enfermedad de Párkinson. La DBS tiene un electrodo que regula la actividad oscilatoria de los ganglios basales involucrados, como el núcleo subtalámico (STN). Un aspecto crítico en el implante de dicho electrodo es la localización precisa de la diana quirúrgica. Esta se realiza mediante imágenes pre-quirúrgicas, marcos estereotácticos y registros de micro-electrodos (MER). Este último permite visualizar la actividad eléctrica de diferentes estructuras a través del recorrido quirúrgico, cada una de ellas con un patrón de variaciones bien definidas en frecuencia; sin embargo, esto dista de ser un método automático o semi-automático que ayude al neurofisiólogo a tomar decisiones en cuanto a la diana quirúrgica. Con el ánimo de contribuir a la automatización, analizamos tres bandas de frecuencias de señales MER adquiridas en 11 pacientes sometidos a DBS: beta (13-40 Hz), gamma (40-200 Hz) y oscilaciones de alta frecuencia (HFO – 201-400 Hz). Se propusieron y evaluaron 5 índices para detectar el STN: variaciones de parámetros auto-regresivos y su derivada a lo largo del recorrido quirúrgico, la energía de cada banda a partir de la densidad espectral de potencia mediante el método de Yule-Walker, la relación de frecuencias altas a bajas y su derivada. Encontramos que la derivada de un parámetro de la banda beta y la relación alta-bajas de las bandas HFO/gamma alcanzaron errores en la localización del STN, similares a los reportados en la literatura (<2mm). Aunque los índices propuestos son sencillos de calcular y de fácil implementación en tiempo real, se deben seguir explorando para incrementar la capacidad de generalización de los resultados obtenidos

Da Vinci robot-assisted radical prostatectomy: initial experience in 50 consecutive cases

Indexación: Web of Science; ScieloObjective: To report our initial experience in 50 cases submitted to a Robotic Radical Prostatectomy (RRP), evaluating results and the learning curve. Material and Methods: From january to october 2010 we performed 50 consecutives cases of RRP with the da Vinci S-HD Surgical System®. The database was performed prospectively, and was analyzed retrospectively. We evaluate demographic data (age, body mass index) and perioperative data such as clinical stage, preoperative PSA (Prostate Specific Antigen), Gleason Score, ASA, operative times, estimated blood loss, morbidity, hospital stay, time of bladder catheterization and positive margins. A statistical analysis of exponential regression was performed to estimate the learning curve. Results: The mean age was 62 years and the most frequent clinical stage was T1c (84%). The mean PSA was 6.36 ng/mL and in 50% of the patients the Gleason Score was 7. The median surgical time was 199 minutes. The mean blood loss was 666 mL (50-4.000 mL). The hospital stay and the average bladder catheterization time were 2 and 6 days, respectively. There were 2 conversions to a laparoscopic approach, none to open surgery, and 8% of postoperative complication (all Clavien 1). Inmediat urinary continence and potency rates were 88.3% and 33.3%, respectively. When comparing the 25 initial cases versus the last 25, there was a decrease in surgical time and estimated blood loss (254 minutes vs 189 minutes and 876 mL vs 467 mL, respectively). We also found a lower rate of positive margins (20% vs 12%). The learning curve statistically estimated is 40 procedures. Conclusion: The surgeon's experience determine a decrease in surgical time, intraoperative bleeding and especially in the rate of positive margins.Objetivo: Comunicar nuestra experiencia inicial en 50 casos de Prostatectomía Radical Robótica (PRR), evaluando resultados y curva de aprendizaje. Material y Métodos: Desde enero a octubre de 2010 se realizaron 50 PRR con el sistema da Vinci S-HD®. La base de datos fue confeccionada en forma prospectiva y se evaluaron en forma retrospectiva los datos demográficos (edad, índice de masa corporal), estadio clínico, valor de Antígeno Prostático Específico (APE), Score de Gleason, ASA, tiempos quirú;rgicos, sangrado estimado, complicaciones, estadía hospitalaria, tiempo de sonda vesical y tasa de márgenes positivos. Se realizó un análisis estadístico de regresión exponencial para estimar la curva de aprendizaje del método. Resultados: La edad media fue de 62 años y el estadio clínico más frecuente fue el T1c (84%). El valor medio de APE fue de 6,36 ng/mL. El score de Gleason en un 50% correspondió al 7 y la media del ASA a 2. La mediana del tiempo quirú;rgico fue de 199 minutos. El sangrado medio estimado fue de 666 mL (50-4.000 mL). La media de la estadía hospitalaria y el tiempo de sonda fueron de 2 y 6 días, respectivamente. Hubo 2 conversiones a cirugía laparoscópica, ninguna a cirugía abierta y un 8% de complicaciones postoperatorias (todas Clavien 1). La tasa de continencia y de potencia inmediata fue de 88,3% y 33,3%, respectivamente. Cuando comparamos los 25 casos iniciales versus los 25 finales hubo un descenso significativo en el tiempo quirú;rgico y sangrado estimado (254 minutos vs 189 minutos y 876 mL vs 467 mL, respectivamente). También encontramos una menor tasa de márgenes positivos en el grupo 2 (12% vs 20%). El análisis estadístico determinó la curva de aprendizaje en 40 procedimientos. Conclusión: Una mayor experiencia del cirujano, determina una disminución en los tiempos quirú;rgicos, sangrado intraoperatorio y sobre todo en la tasa de márgenes positivos.http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-40262011000600011&nrm=is

Gene discovery for facioscapulohumeral muscular dystrophy by machine learning techniques

Facioscapulohumeral muscular dystrophy (FSHD) is a neuromuscular disorder that shows a preference for the facial, shoulder and upper arm muscles. FSHD affects about one in 20-400,000 people, and no effective therapeutic strategies are known to halt disease progression or reverse muscle weakness or atrophy. Many genes may be incorrectly regulated in affected muscle tissue, but the mechanisms responsible for the progressive muscle weakness remain largely unknown. Although machine learning (ML) has made significant inroads in biomedical disciplines such as cancer research, no reports have yet addressed FSHD analysis using ML techniques. This study explores a specific FSHD data set from a ML perspective. We report results showing a very promising small group of genes that clearly separates FSHD samples from healthy samples. In addition to numerical prediction figures, we show data visualizations and biological evidence illustrating the potential usefulness of these results.Peer ReviewedPostprint (published version

Proinflammatory and Anabolic Gene Expression Effects of Platelet-Rich Gel Supernatants on Equine Synovial Membrane Explants Challenged with Lipopolysaccharide

Platelet-rich plasma (PRP) preparations are used in horses with osteoarthritis (OA). However, some controversies remain regarding the ideal concentration of platelets and leukocytes to produce an adequate anti-inflammatory and anabolic response in the synovial membrane. The aims of this study were to study the influence of leukoconcentrated platelet-rich gel (Lc-PRG) and leukoreduced platelet-rich gel (Lr-PRG) supernatants on the quantitative expression of some proinflammatory and anabolic genes in equine synovial membrane explants (SMEs) challenged with lipopolysaccharide (LPS). SMEs from six horses were cultured over 96 h. Then, SMEs were harvested for RNA extraction and quantitative gene expression analysis by RT-qPCR for nuclear factor kappa B (NFκB), matrix metalloproteinase 13 (MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4), collagen type I alpha 1 (COL1A1), collagen type II alpha 1 (COL2A1), and cartilage oligomeric matrix protein (COMP). The 25% and 50% Lc-PRG supernatants led to downregulation of NFκB, MMP-13, ADAMTS-4, COL1A1, COL2A1, and COMP in SMEs. Lr-PRG supernatants (particularly at the 50% concentration) induced downregulation of NFκB, MMP-13, ADAMTS-4, and COL1A1 and upregulation of COL2A1 and COMP. Lr-PRG supernatants should be used for the treatment of inflammatory arthropathies in horses because they have anti-inflammatory and anabolic effects in the synovial membrane