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The inhibition of glycogen synthase kinase-3 (GSK-3) can induce neurogenesis, and the associated activation of Wnt/β-catenin signaling via GSK-3 inhibition may represent a means to promote motor function recovery following spinal cord injury (SCI) via increased astrocyte migration, reduced astrocyte apoptosis, and enhanced axonal growth. Herein, we assessed the effects of GSK-3 inhibition in vitro on the neurogenesis of ependymal stem/progenitor cells (epSPCs) resident in the mouse spinal cord and of human embryonic stem cell-derived neural progenitors (hESC-NPs) and human-induced pluripotent stem cell-derived neural progenitors (hiPSC-NPs) and in vivo on spinal cord tissue regeneration and motor activity after SCI. We report that the treatment of epSPCs and human pluripotent stem cell-derived neural progenitors (hPSC-NPs) with the GSK-3 inhibitor Ro3303544 activates β-catenin signaling and increases the expression of the bIII-tubulin neuronal marker; furthermore, the differentiation of Ro3303544-treated cells prompted an increase in the number of terminally differentiated neurons. Administration of a water-soluble, bioavailable form of this GSK-3 inhibitor (Ro3303544-Cl) in a severe SCI mouse model revealed the increased expression of bIII-tubulin in the injury epicenter. Treatment with Ro3303544-Cl increased survival of mature neuron types from the propriospinal tract (vGlut1, Parv) and raphe tract (5-HT), protein kinase C gamma-positive neurons, and GABAergic interneurons (GAD65/67) above the injury epicenter. Moreover, we observed higher numbers of newly born BrdU/DCX-positive neurons in Ro3303544-Cl-treated animal tissues, a reduced area delimited by astrocyte scar borders, and improved motor function. Based on this study, we believe that treating animals with epSPCs or hPSC-NPs in combination with Ro3303544-Cl deserves further investigation towards the development of a possible therapeutic strategy for SCI.

The human iPSC cell line, ARS-FiPS4F1 (ESi063-A), derived from dermal fibroblast from the patient autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) caused by mutations on the gene SACSIN, was generated by non-integrative reprogramming technology using OCT3/4, SOX2, CMYC and KLF4 reprogramming factors. The pluripotency was assessed by immunocytochemistry and RT-PCR. Differentiation capacity was verified in vitro. This iPSC line can be further differentiated toward affected cells to better understand molecular mechanisms of disease and pathophysiology.

Cerebellar ataxias are clinically and genetically heterogeneous diseases affecting primary cerebellar cells. The lack of availability of affected tissue from cerebellar ataxias patients is the main obstacle in investigating the pathogenicity of these diseases. The landmark discovery of human-induced pluripotent stem cells (hiPSC) has permitted the derivation of patient-specific cells with an unlimited self-renewing capacity. Additionally, their potential to differentiate into virtually any cell type of the human organism allows for large amounts of affected cells to be generated in culture, converting this hiPSC technology into a revolutionary tool in the study of the mechanisms of disease, drug discovery, and gene correction. In this review, we will summarize the current studies in which hiPSC were utilized to study cerebellar ataxias. Describing the currently available 2D and 3D hiPSC-based cellular models, and due to the fact that extracerebellar cells were used to model these diseases, we will discuss whether or not they represent a faithful cellular model and whether they have contributed to a better understanding of disease mechanisms.

Background and Purpose— Animal models of transient ischemia suggest that oxygen-derived free radicals produced on reperfusion of ischemic brain could constitute the main cause of reperfusion injury. We aimed to determine the presence and role of lipid peroxidation and protein oxidation-related molecules after tissue plasminogen activator-induced recanalization in human stroke. Methods— A total of 160 patients with strokes involving the middle cerebral artery and treated with tissue plasminogen activator and 60 healthy controls were included. Blood samples, transcranial Doppler recordings, and National Institutes of Health Stroke Scale scores were obtained at baseline (pretreatment), 1 hour and 2 hours after tissue plasminogen activator bolus, and 12 hours and 24 hours after stroke onset. The main lipid peroxidation end-product malondialdehyde, advanced oxidation protein products, and plasma concentrations of myeloperoxidase were assessed. Results— At baseline, all oxidative stress biomarkers were higher than in control subjects ( P Conclusions— Our study showed that systemic oxidative damage to lipids and proteins had already occurred at baseline in stroke. In contrast to animal studies, a relationship between free radical-mediated oxidative damage to lipids or proteins and reperfusion injury after arterial recanalization could not be established.

Background and goal of study: there is evidence that perioperative intravenous ketamine and lidocaine reduce postoperative pain, postoperative opioids consumption, shortens hospital stay and accelerates intestinal function recovery. However, it has not been studied the beneficial effects in the intraoperative period. The aim of this study was to evaluate the effect of a single dose of lidocaine and ketamine on intraoperative opioids requirements in patients undergoing elective gynecological laparotomies under general anesthesia.we performed a single-centre, prospective, randomized, double-blinded, placebo-controlled study. We included 33 patients (11 in the ketamine group, 11 in the lidocaine group and 11 in the placebo group). Postoperative analgesia was accomplished by patient-controlled morphine. Patients were randomly assigned to receive either a 1.5 mg/kg of 2% lidocaine, 0.5 mg/kg of 5% ketamine or 0.9% saline bolus. The primary outcome was the opioids consumption during surgery. The secondary outcomes included: emergence time, pain scores, opioids consumption within 24 h after surgery and side effects.decreased intraoperative opioids requirements were noted in the experimental groups (ketamine: 402.3 } 106.3 and lidocaine: 397.7 } 107.5, compared with saline: 561.4 } 97.1); p = 0.001. We found a positive correlation between intraoperative opioids consumption and emergence time (r = 0.864, p0.001). There was no significant difference between the groups in VAS pain scores at rest within the first 24 postoperative hours. Total morphine consumption within 24 h after surgery did not differ significantly among the groups (placebo: 27.54 } 11.75; ketamine: 30.95 } 7.88; lidocaine 34.77 } 4510.25; p = 0.26). Postoperative nausea and vomiting were more common in placebo group (it was observed in 3 subjects in ketamine group, in 5 subjects in lidocaine group and in 9 subjects in placebo group; p = 0.027).our results do not support the use of intraoperative single dose of lidocaine or ketamine to reduce postoperative pain and postoperative opioids consumption after open gynecological surgery. However, they seem to decrease intraoperative opioids requirements and shorten emergence time. Nevertheless, these findings should be validating in further studies with large sample size.Introducción y objetivos del estudio: existe evidencia de que la administracion perioperatoria de ketamina y lidocaina intravenosa reduce el dolor y el consumo de opioides postoperatorio, acorta la estancia hospitalaria y acelera la recuperacion de la funcion intestinal. Sin embargo, no se han estudiado los efectos beneficiosos en el periodo intraoperatorio. El objetivo de este estudio fue evaluar el efecto de una unica dosis de lidocaina y ketamina sobre el consumo intraoperatorio de opioides en pacientes sometidas a cirugia ginecologica electiva bajo anestesia general. Material y métodos: estudio prospectivo, aleatorizado, doble ciego, controlado con placebo en un solo centro. Se incluyeron 33 pacientes (11 en el grupo ketamina, 11 en el grupo lidocaina y 11 en el grupo placebo). Para la analgesia postoperatoria se utilizo una bomba PCA (Analgesia Controlada por el Paciente ) de morfina. Los pacientes fueron asignados al azar a uno de los tres grupos de estudio: 1,5 mg/kg de lidocaina al 2%, 0,5 mg/kg de ketamina al 5% o solucion salina 0.9%. La variable principal del estudio fue el consumo de opioides durante la cirugia. Las variables secundarias fueron: tiempo de educcion de la anestesia, intensidad del dolor, consumo de opioides en las 24 horas posteriores a la cirugia y efectos adversos. Resultados: se observo una disminucion del consumo intraoperatorio de opioides en los grupos ketamina (402,3 } 106,3) y lidocaina (397,7 } 107,5) frente al grupo placebo (561,4 } 97,1); p = 0,001. Se encontro una correlacion positiva entre el consumo intraoperatorio de opioides y el tiempo de despertar (r = 0,864, p.

Down’s syndrome (DS) is characterized by a complex phenotype associated with chronic oxidative stress and mitochondrial dysfunction. Overexpression of genes on chromosome-21 is thought to underlie the pathogenesis of the major phenotypic features of DS, such as premature aging. Using cultured fibroblasts with trisomy 21 (T21F), this study aimed to ascertain whether an imbalance exists in activities, mRNA, and protein expression of the antioxidant enzymes SOD1, SOD2, glutathione-peroxidase, and catalase during the cell replication processin vitro. T21F had high SOD1 expression and activity which led to an interenzymatic imbalance in the antioxidant defense system, accentuated with replicative senescence. Intracellular ROS production and oxidized protein levels were significantly higher in T21F compared with control cells; furthermore, a significant decline in intracellular ATP content was detected in T21F. Cell senescence was found to appear prematurely in DS cells as shown by SA-β-Gal assay and p21 assessment, though not apoptosis, as neither p53 nor the proapoptotic proteins cytochrome c and caspase 9 were altered in T21F. These novel findings would point to a deleterious role of oxidatively modified molecules in early cell senescence of T21F, thereby linking replicative and stress-induced senescence in cultured cells to premature aging in DS.

Background and Purpose— Fluorescent molecular peroxidation products (FMPPs) are considered potential markers of molecular oxidative damage and may provoke increased permeability and disruption of the blood–brain barrier. This study aimed to determine the value of FMPPs as a biomarker to predict neurological worsening related to early hemorrhagic transformation. Methods— Baseline FMPP levels were measured in 186 consecutive acute ischemic stroke patients before tissue plasminogen activator treatment was administered. A serial FMPP profile (baseline before tissue plasminogen activator treatment, and 1, 2, 12, and 24 hours from treatment) was determined in a subset of 100 patients. Computed tomographic scans were performed at admission and repeated at 24 to 48 hours or after neurological worsening occurred. Symptomatic intracranial hemorrhage was defined as blood at any site in the brain associated with neurological deterioration. Results— Patients who worsened had higher median FMPP levels compared with those who did not (59.68 [48.63–85.73] versus 44.87 [36.37–58.90] Uf/mL; P =0.035) at baseline. After logistic regression multivariate analysis, FMPP >48.2 Uf/mL together with age, hypertension, and systolic blood pressure remained baseline predictors of worsening at 48 hours. Moreover, baseline FMPP determination helped to distinguish between patients who worsened and those who did not (Integrated Discrimination Improvement index, 5.7%; P =0.0004). Finally, within patients who had worsened at 48 hours, those with symptomatic intracranial hemorrhage had higher FMPP levels ( P =0.038). Conclusions— FMPPs might be a valuable biomarker of poor early neurological outcome and be related to the appearance of symptomatic intracranial hemorrhage in tissue plasminogen activator–treated patients, one of the most feared neurological complications after thrombolytic treatment of acute ischemic stroke.

To assess the profile of patients receiving antibiotics and the appropriateness of these prescriptions for the clinical conditions.Cross-sectional study of prescription-indication.A primary health care area in Andalusia.Patients assigned to primary care centres. Patients with antibiotic prescriptions during 2009 were selected by simple random sampling (confidence level: 95%, accuracy: 5%). Primary endpoint: appropriateness of antibiotics prescribing to recommendations included in local guidelines. Data were obtained through the billing computerised prescriptions system and medical histories.Twenty-five per cent of the population area received antibiotics during 2009. The 1,266 patient samples showed the following characteristics: 57.9% were women, with a mean age of 41 (±1) years. There were 39.3% pensioners. The percentage of appropriate antibiotic prescriptions was 19.9%, with no difference due to gender. Statistically significant differences were related to age, being those over 65 years the group of patients with the highest percentage of inappropriateness. The main reasons for inappropriateness were: no recording of the infectious process (44.5%), a wrong treatment duration (15.5%), and the use of an inadequate antibiotic (11.5%).There is a high level of inappropriateness in antibiotic prescribing in primary care. The high level of under-recording of diagnoses, mainly in elderly patients, followed by the use of wrong schedules, and the wrong type of antibiotics were the main reasons of inappropriateness.

Objective To investigate the impact of early cleavage (EC) on embryo quality, implantation, and live-birth rates. Design Prospective cross-sectional study. Setting Multicenter study. Patient(s) Seven hundred embryo transfers and 1,028 early-stage human embryos. Intervention(s) None. Main Outcome Measure(s) Implantation according to the presence of EC and embryo quality. Result(s) The presence of EC is associated with embryo quality, especially in cycles with autologous oocytes. However, the use of EC as an additional criterion for selecting an embryo for transfer does not appear to significantly improve likelihood of implantation. Furthermore, embryos that presented EC had live-birth rates per implanted embryo similar to those that did not show any sign of cleavage. Conclusion(s) At least for conventional embryo culture and morphologic evaluations, the additional evaluation of EC in embryos may not be valuable to improve embryo implantation.

Background and aims Chitotriosidase, a component of innate immunity, constitutes a sensitive parameter of macrophage activation and its elevated plasma activity reflects an inflammatory response. Given the deleterious effects of inflammation in brain ischemia, we aimed to assess the prognostic value of chitotriosidase activity in acute stroke patients. Methods The study comprised 159 acute stroke patients and 51 age-matched controls. Plasma chitotriosidase activity was serially determined by fluorometric assay. Short-term neurological outcome was determined at 48 h and functional outcome at three-months. Predictors of neurological and functional outcome were determined via multivariate analysis, and the additional predictive value of chitotriosidase was tested with the Integrated Discrimination Index and the Net Reclassification Improvement. Results Stroke patients showed increased levels of baseline chitotriosidase activity compared to controls [114·2 (74·65–182·95) nmol/ml/h vs. 54·4 (32·7–76·4); P < 0·0001]. Chitotriosidase activity (Conclusions Baseline chitotriosidase activity in acute stroke patients treated with tissue plasminogen activator (tPA) may constitute a prognostic predictor of short-term outcome, adding a moderate additional predictive value. Our results underline the deleterious role of inflammation in acute stroke patients.

Several soft-tissue dermal fillers have been reported to provoke immunogenicity and may cause adverse reactions despite claims regarding their safety. This study aimed to assess biomaterial-induced macrophage activation, cell-mediated immune response and oxidative stress in 169 patients with dermal bioimplants. To this end, we analysed plasma concentrations of myeloperoxidase (MPO), the chitinase-like proteins chitotriosidase and YKL-40 and molecular oxidative damage. The present study shows, for the first time, that the components of innate immunity: chitotriosidase and YKL-40, are significantly higher in patients with certain bioimplants and these markers of monocyte/macrophage activation rose progressively as adverse reactions (AR) evolved. Plasma MPO levels increased 4-fold in filler users with AR and 3-fold in those without. Analysis by filler type showed subjects injected with calcium hydroxylapatite, methacrylate, acrylamides and silicone to have values significantly above those of non-filler subjects for at least two plasma biomarkers, probably because the afore-mentioned biomaterials are permanent and prone to trigger AR in the long term. By contrast, hyaluronic acid alone elicited little immune response. Plasma concentrations of markers of oxidative damage to lipids and proteins were found to be significantly higher in users of four of the nine dermal fillers studied. These diffusible products of molecular peroxidation would stem from the reaction catalysed by MPO that generates potent oxidants, leading to cell oxidative damage which, in turn, may exert deleterious effects on the organism. Overall, the results of this study on the effects of a range of dermal fillers point to chronic activation of the immune response mediated by macrophages and PMNs. The increases in plasma of MPO, chitotriosidase and YKL-40 proteins and products of macromolecular peroxidation suggests that these molecules could serve as blood-based biochemical markers and alert to the risk of chronic immune system activation and development of adverse events that may arise from the use of certain bioimplants.

A recent study has shown that chitotriosidase (Chit) may play a role in the pathogenesis of multiple sclerosis (MS). Plasma Chit activity was investigated in 219 untreated MS patients and 160 healthy controls (HC) by means of a fluorometric enzyme activity assay. Chit activity was also measured in a subgroup of 46 patients following treatment with interferon-beta (IFNbeta). Overall, plasma Chit activity was significantly increased in MS patients compared with HC, but no differences were observed between relapsing and progressive clinical forms. In addition, Chit activity was similar between patients during relapse and patients during clinical remission. Treatment with IFNbeta was associated with a significant increase in Chit activity compared with untreated patients in both responders and non-responders to treatment. Although these findings suggest a role of Chit in MS, our data do not support an association between plasma Chit activity and MS clinical course and clinical response to IFNbeta treatment.