Differential Association of 4E-BP2-Interacting Proteins Is Related to Selective Delayed Neuronal Death after Ischemia.

Cerebral ischemia induces an inhibition of protein synthesis and causes cell death and neuronal deficits. These deleterious effects do not occur in resilient areas of the brain, where protein synthesis is restored. In cellular stress conditions, as brain ischemia, translational repressors named eukaryotic initiation factor (eIF) 4E-binding proteins (4E-BPs) specifically bind to eIF4E and are critical in the translational control. We previously described that 4E-BP2 protein, highly expressed in brain, can be a molecular target for the control of cell death or survival in the reperfusion after ischemia in an animal model of transient cerebral ischemia. Since these previous studies showed that phosphorylation would not be the regulation that controls the binding of 4E-BP2 to eIF4E under ischemic stress, we decided to investigate the differential detection of 4E-BP2-interacting proteins in two brain regions with different vulnerability to ischemia-reperfusion (IR) in this animal model, to discover new potential 4E-BP2 modulators and biomarkers of cerebral ischemia. For this purpose, 4E-BP2 immunoprecipitates from the resistant cortical region and the vulnerable hippocampal cornu ammonis 1 (CA1) region were analyzed by two-dimensional (2-D) fluorescence difference in gel electrophoresis (DIGE), and after a biological variation analysis, 4E-BP2-interacting proteins were identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Interestingly, among the 4E-BP2-interacting proteins identified, heat shock 70 kDa protein-8 (HSC70), dihydropyrimidinase-related protein-2 (DRP2), enolase-1, ubiquitin carboxyl-terminal hydrolase isozyme-L1 (UCHL1), adenylate kinase isoenzyme-1 (ADK1), nucleoside diphosphate kinase-A (NDKA), and Rho GDP-dissociation inhibitor-1 (Rho-GDI), were of notable interest, showing significant differences in their association with 4E-BP2 between resistant and vulnerable regions to ischemic stress. Our data contributes to the first characterization of the 4E-BP2 interactome, increasing the knowledge in the molecular basis of the protection and vulnerability of the ischemic regions and opens the way to detect new biomarkers and therapeutic targets for diagnosis and treatment of cerebral ischemia.

Stress Granule Induction after Brain Ischemia Is Independent of Eukaryotic Translation Initiation Factor (eIF) 2α Phosphorylation and Is Correlated with a Decrease in eIF4B and eIF4E Proteins.

Stress granules (SGs) are cytoplasmic ribonucleoprotein aggregates that are directly connected with the translation initiation arrest response to cellular stresses. Translation inhibition (TI) is observed in transient brain ischemia, a condition that induces persistent TI even after reperfusion, i.e. when blood flow is restored, and causes delayed neuronal death (DND) in selective vulnerable regions. We previously described a connection between TI and DND in the hippocampal cornu ammonis 1 (CA1) in an animal model of transient brain ischemia. To link the formation of SGs to TI and DND after brain ischemia, we investigated SG induction in brain regions with differential vulnerabilities to ischemia-reperfusion (IR) in this animal model. SG formation is triggered by both eukaryotic translation initiation factor (eIF) 2α phosphorylation and eIF4F complex dysfunction. We analyzed SGs by immunofluorescence colocalization of granule-associated protein T-cell internal antigen-1 with eIF3b, eIF4E, and ribosomal protein S6 and studied eIF2 and eIF4F complex. The results showed that IR stress induced SG formation in the CA1 region after 3-day reperfusion, consistent with TI and DND in CA1. SGs were formed independently of eIF2α phosphorylation, and their appearance was correlated with a decrease in the levels of eIF4F compounds, the cap-binding protein eIF4E, and eIF4B, suggesting that remodeling of the eIF4F complex was required for SG formation. Finally, pharmacological protection of CA1 ischemic neurons with cycloheximide decreased the formation of SGs and restored eIF4E and eIF4B levels in CA1. These findings link changes in eIF4B and eIF4E to SG induction in regions vulnerable to death after IR.

Thalamic deep brain stimulation for orthostatic tremor: A multicenter international registry.

BACKGROUND: We report the accumulated experience with ventral intermediate nucleus deep brain stimulation for medically refractory orthostatic tremor. METHODS: Data from 17 patients were reviewed, comparing presurgical, short-term (0-48 months), and long-term (≥48 months) follow-up. The primary end point was the composite activities of daily living/instrumental activities of daily living score. Secondary end points included latency of symptoms on standing and treatment-related complications. RESULTS: There was a 21.6% improvement (P = 0.004) in the composite activities of daily living/instrumental activities of daily living score, which gradually attenuated (12.5%) in the subgroup of patients with an additional long-term follow-up (8 of 17). The latency of symptoms on standing significantly improved, both in the short-term (P = 0.001) and in the long-term (P = 0.018). Three patients obtained no/minimal benefit from the procedure. CONCLUSIONS: Deep brain stimulation of the ventral intermediate nucleus was, in general, safe and well tolerated, yielding sustained benefit in selected patients with medically refractory orthostatic tremor. © 2017 International Parkinson and Movement Disorder Society.

Usefulness of the H-Reflex for Intraoperative Monitoring of Thoracoabdominal Aneurysms.

PURPOSE: Intraoperative neurophysiologic monitoring in thoracoabdominal aneurysms (TAAA) is essential to avoid intraoperative spinal cord injury). Motor and somatosensory evoked potentials may be considered intraoperative tools for detecting spinal cord injury. H-reflex is a well-known neurophysiologic technique to evaluate L5-S1 root. Current evidence supports the observation that H-reflex changes may occur with spinal cord damage as high as the cervical level. This study aimed to evaluate the usefulness of the H-reflex in these surgeries. METHODS: The use of intraoperative H-reflex in TAAA monitoring was evaluated in 12 patients undergoing open or endovascular repair of TAAA for a period of four years (2016-2020) using somatosensory evoked potentials (SSEPs) and transcranial motor evoked potentials (TcMEPs) and bilateral H-reflex. RESULTS: Six neurophysiologic alarms were recorded in five of the 12 patients. Summarizing the neurophysiologic changes of our series, we considered a peripheral change when we detected a unilateral loss of SSEPs, TcMEPs, and H-reflex. Instead, we assumed a central change when we detected a unilateral or bilateral loss of TcMEPs and H-reflex with normal SSEPs, which we considered a sign of spinal cord ischemia. Interestingly H-reflex always changed significantly in combination with TcMEPs in the same fashion. CONCLUSIONS: According to our series, H-reflex can detect intraoperative changes with the same sensitivity as TcMEPs in TAAA surgeries. With the support of other techniques, it can be useful to localize the origin of the lesion (peripheral or central spinal cord), to help in surgical decision-making to avoid postoperative neurologic damage. Based on our results, we recommend the systematic use of H-reflex in TAAA surgeries.

Combined Stimulation of the Substantia Nigra and the Subthalamic Nucleus for the Treatment of Refractory Gait Disturbances in Parkinson's Disease: A Preliminary Study.

Deep brain stimulation of the subthalamic nucleus is efficient for the treatment of motor symptoms (i.e., tremors) in patients with Parkinson's disease. Gait disorders usually appear during advanced stages of idiopathic Parkinson's disease in up to 80% of patients and have an important impact on their quality of life. The effects of deep brain stimulation of the subthalamic nucleus on gait and balance are still controversial. For this reason, alternative targets have been considered, such as stimulation of the pedunculopontine nucleus and the pars reticulata of substantia nigra, involved in the integration of the functional connections for gait. Due to the proximity of the subthalamic nucleus to the substantia nigra, their combined stimulation is feasible and may lead to better outcomes, improving axial symptoms. Our objective was to prospectively compare simultaneous stimulation of both structures versus conventional subthalamic stimulation in improving gait disorders. In ten patients with advanced Parkinson's disease, deep brain stimulation leads (eight linear contacts) were implanted, and gait analysis was performed 6 months after surgery in off-stimulation and after 4 weeks of dual or single subthalamic stimulation. An improvement in gait parameters was confirmed with both stimulation conditions, with better results with combined substantia nigra and subthalamic stimulation compared with conventional subthalamic stimulation. Further studies are needed to determine if this effect remains after long-term dual-target stimulation.

Pulse-train versus single-pulse t-EMG stimulation for intraoperative neurophysiologic monitoring of thoracic pedicle screws in adolescent idiopathic scoliosis.

OBJECTIVE: To assess the PTS method in neuromonitoring of thoracic pedicle screws in a young cohort of AIS patients. The accuracy of both PTS (Pulse-train stimulation) and SPS (Single-pulse stimulation) techniques in the detection of misplaced thoracic screws was compared with special reference to the screws placed at the concavity and the convexity of the curve. PATIENTS AND METHODS: A single-center prospective clinical cohort study. LEVEL OF EVIDENCE II: Twelve AIS patients who underwent elective surgery were included in this study. Screw stimulation using repetitive constant-current stimulus train of variable intensities was first performed after screw insertion. SPS was performed immediately after PTS. Postoperatively, CT scan was used to check the final position of the screws. A total of 246 thoracic pedicle screws were placed. Thresholds of screws placed in the convexity and the concavity were compared. RESULTS: Invasion of the spinal canal was postoperatively confirmed in 29 of the pedicle screws. The SPS technique detected three (10.3%) of these screws using a threshold limit of 12 mA. The PTS technique detected 25 (86.2%) of the 29 misplaced screws using threshold of 30 mA (negative predictive value, 93.1%). When using a PTS threshold of < 20 mA, the positive predictive value was 70%. SPS did not detect any of the misplaced screws at the apex level of the scoliotic curves, and PTS detected 10 out of 17 of these screws. Overall, area under the ROC curve was 0.82 for PTS and 0.61 for SPS. CONCLUSIONS: PTS is a reliable method for detecting medially misplaced screws among young AIS patients undergoing scoliosis surgery. PTS provides more accurate predictions than SPS and improves the identification of screws invading the spinal canal at the apex or near the apex levels.

Effects of deep brain stimulation on the kinematics of gait and balance in patients with idiopathic Parkinson's disease.

BACKGROUND: Advanced stages of idiopathic Parkinson's disease are often characterised by gait alterations and postural instability. Despite improvements in patients' motor symptoms after deep brain stimulation of the subthalamic nucleus, its effects on gait and balance remain a matter of debate. This study investigated the effects of deep brain stimulation on balance and kinematic parameters of gait. METHODS: The gait of 26 patients with advanced idiopathic Parkinson's disease was analysed before and after (between 3 and 6 months) after bilateral deep brain stimulation of the subthalamic nucleus. Computerised analysis was used to study cadence, number of cycles with the correct support sequence, number of cycles, duration of the cycle stages, and knee and ankle goniometry. Balance, postural instability, and mobility were assessed using the Tinetti and Timed Up and Go test. FINDINGS: After stimulation, the following changes were significant (p < 0.01): number of cycles with the correct support sequence, number of total cycles, and foot contact. Patients improved significantly (p < 0.01) in the Tinetti and Timed Up and Go tests, the risk factors for falls changed from high (median 17) to low (median 25), and they improved from minor dependence (statistical median 14) to normality (statistical median 8.70). INTERPRETATION: Deep brain stimulation to inhibit hyperactivity of the subthalamic nucleus was associated with an improvement in the space-time variables of gait and balance in patients with Parkinson's disease for up to 3-6 months. These results highlight the major role of the subthalamic nucleus in motor control mechanisms during locomotion and balance.

Recording triggered EMG thresholds from axillary chest wall electrodes: a new refined technique for accurate upper thoracic (T2-T6) pedicle screw placement.

This study was aimed at evaluating the sensitivity and safety of a new technique to record triggered EMG thresholds from axillary chest wall electrodes when inserting pedicle screws in the upper thoracic spine (T2-T6). A total of 248 (36.6%) of a total of 677 thoracic screws were placed at the T2-T6 levels in 92 patients with adolescent idiopathic scoliosis. A single electrode placed at the axillary midline was able to record potentials during surgery from all T2-T6 myotomes at each side. Eleven screws were removed during surgery because of malposition according to intraoperative fluoroscopic views. Screw position was evaluated after surgery in the remaining 237 screws using a CT scan. Malposition was detected in 35 pedicle screws (14.7%). Pedicle medial cortex was breached in 24 (10.1%). Six screws (2.5%) were located inside the spinal canal. Mean EMG threshold was 24.44 ± 11.30 mA in well-positioned screws, 17.98 ± 8.24 mA (p < 0.01) in screws violating the pedicle medial cortex, and 10.38 ± 3.33 mA (p < 0.005) in screws located inside the spinal canal. Below a threshold of 12 mA, 33.4% of the screws (10/30) were malpositioned. Furthermore, 36% of the pedicle screws with t-EMG stimulation thresholds within the range 6-12 mA were malpositioned. In conclusion, assessment of upper thoracic pedicle screw placement by recording tEMG at a single axillary electrode was highly reliable. Thresholds below 12 mA should alert surgeons to suspect screw malposition. This technique simplifies tEMG potential recording to facilitate safe placement of pedicle screws at upper thoracic levels.

Neuromuscular involvement in COVID-19 critically ill patients.

OBJECTIVE: Coronavirus disease 2019 (COVID-19) has a high incidence of intensive care admittance due to the severe acute respiratory syndrome (SARS). Intensive care unit (ICU)-acquired weakness (ICUAW) is a common complication of ICU patients consisting of symmetric and generalised weakness. The aim of this study was to determine the presence of myopathy, neuropathy or both in ICU patients affected by COVID-19 and whether ICUAW associated with COVID-19 differs from other aetiologies. METHODS: Twelve SARS CoV-2 positive patients referred with the suspicion of critical illness myopathy (CIM) or polyneuropathy (CIP) were included between March and May 2020. Nerve conduction and concentric needle electromyography were performed in all patients while admitted to the hospital. Muscle biopsies were obtained in three patients. RESULTS: Four patients presented signs of a sensory-motor axonal polyneuropathy and seven patients showed signs of myopathy. One muscle biopsy showed scattered necrotic and regenerative fibres without inflammatory signs. The other two biopsies showed non-specific myopathic findings. CONCLUSIONS: We have not found any distinctive features in the studies of the ICU patients affected by SARS-CoV-2 infection. SIGNIFICANCE: Further studies are needed to determine whether COVID-19-related CIM/CIP has different features from other aetiologies. Neurophysiological studies are essential in the diagnosis of these patients.

Expression of heat shock protein 90 at the cell surface in human neuroblastoma cells.

In addition to the activity of heat shock protein 90 (Hsp90/HSPC) as a chaperone, some recent studies have reported expression of Hsp90 at the cell surface in certain types of cancer and nervous system cells. We study the expression of Hsp90 at the cell surface in human neuroblastoma (NB69) cells. Immunofluorescence experiments labeling with anti-Hsp90 antibodies on both nonpermeabilized cells and live cells detected Hsp90 at the cell surface. Hsp90 was also identified in a membrane fraction from subcellular fractionation. Cell-surface Hsp90 was significantly more expressed in undifferentiated proliferative spherical neuroblastoma cells than in differentiated flattened cells. In addition, spherical cells were significantly more sensitive to Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin compared to flattened cells. This paper describes the first evidence of cell-surface Hsp90 expression in a cancer cell line from nervous tissue and may indicate a novel target for anti-tumoral agents.

[Intraoperative neurophysiological monitoring in radical prostatectomy and pudendal nerve surgical releasing.] / Monitorización neurofisiológica intraoperatoria en prostatectomía radical y liberación del nervio pudendo.

OBJECTIVE: Intraoperative neurophysiological monitoring (INM) allows obtaining real-time information on the functional integrity of nervous system structures. The objective of this article is to present the role of clinical neurophysiology in Urology in the identification and preservation, on the one hand, of the prostate neurovascular bundles in radical prostatectomy (RP), and of the pudendal nerve in the release of this in Pudendal Nerve Entrapment Syndrome (PNS). METHODS: A bipolar laparoscopic probe was used for intraoperative stimulation for both the identification in PNS and neurovascular bundles in RP, obtaining response at the external anal sphincter in the first one; and intracavernous pressure in the second through needle electrodes. RESULTS: Preservation of the periprostatic neurovascular bundles allows to increase rates of sexual potency and urinary continence after surgery. However, it has been shown that the innervation of the corpora cavernosa and the urinary sphincter has a variable disposition. Intraoperative neurostimulation allows the most precise identification and dissection of the vascular and nervous structures that surround the prostate. In the diagnosis of PNS, the neurophysiological study allows to rule out pathology at other levels, such as, primary muscular pathology of the anal sphincter. There are no pathognomonic neurophysiological findings of PNS. In addition, it allows the identification of the nerve during the laparoscopic release of the nerve at the entrance of the Alcock channel, as well as the evaluation of its functional integrity after the end of the dissection. CONCLUSIONS: The INM is a fundamental tool to allow an improved identification of nerve structures during RP and PNS in order to preserve them. It also helps with the diagnosis of PNS.

OBJETIVO: La monitorización neurofisiológica intraoperatoria (MNI) permite obtener información en tiempo real sobre la integridad funcional de las estructuras del sistema nervioso. El objetivo de este trabajo exponer el papel de neurofisiología clínica en Urología en la identificación y preservación, por un lado, de los haces neurovasculares prostáticos en la prostatectomía radical (PR), y del nervio pudendo en la liberación de este en el Síndrome de Atrapamiento del Nervio Pudendo (SANP).MÉTODOS: Se empleó una sonda bipolar laparoscópica para la estimulación intraoperatoria tanto para la identificación en SANP como de haces neurovasculares en PR obteniéndose respuesta a nivel del esfínter anal en la primera y presión intracavernosa en la segunda por medio de electrodos de aguja. RESULTADOS: La presentación de los haces neurovasculares periprostáticos permite aumentar las tasas de potencia sexual y continencia urinaria. Si bien, posteriormente, se evidenció que la inervación de los cuerpos cavernosos y el esfínter urinario tiene disposición variable. La neuroestimulación intraoperatoria permite la identificación y disección más precisa de las estructuras vasculonerviosas que circundan la próstata. En el diagnóstico del SANP, el estudio neurofisiológico permite descartar patología a otros niveles, como por ejemplo patología primaria muscular del esfínter anal; así como, valorar las ramas del nervio pudendo. No existen hallazgos neurofisiológicos patognomónicos del SANP. Además, permite la identificación del nervio durante la liberación laparoscópica del nervio a la entrada del canal de Alcock, así como la evaluación de su integridad funcional tras finalizar la disección. CONCLUSIONES: La MNI es una herramienta fundamental para permitir una mejora en la identificación de estructuras nerviosas durante PR y SANP para poder preservarlas. Así mismo, ayuda con el diagnóstico de SANP.

Impact of Disease Duration in Effectiveness of Treatment with Levodopa-Carbidopa Intestinal Gel and Factors Leading to Discontinuation.

BACKGROUND: Levodopa-carbidopa intestinal gel (LCIG) is effective in the treatment of advanced Parkinson's disease (PD). However, the patients' profile that might benefit from treatment with LCIG has not been characterized. OBJECTIVE: This retrospective study explored the influence of disease duration (DD) on the effectiveness of LCIG and identified factors associated with treatment discontinuation in a cohort of advanced PD patients. METHODS: Patients initiating LCIG therapy between Jan-2006 and Dec-2011 in 18 Spanish centers were included. Effectiveness in treating motor symptoms (MSs), non-motor symptoms (NMSs), and adverse events (AEs) occurrence was compared in DD≥10 or <10 years and LCIG continuation/discontinuation groups. Factors associated with LCIG discontinuation were evaluated using univariate and multivariate analyses. RESULTS: Overall, 177 PD patients were included (52.5% male; mean age 70.6±8.4 years; mean LCIG duration 35.6±18.6 months). Patients with DD≥10 years (n = 125) experienced less reduction in "off" time (-29%) than those with DD <10 years (-38%; n = 51; p = 0.021), and reported more severe AEs (32.8% vs. 17.6%; p = 0.043). DD did not significantly influence changes in NMSs or discontinuation rates. Fifty-four patients discontinued LCIG therapy, factors associated with discontinuation were higher percentages of waking day in the "off" state (OR, 1.028; 95% CI, 1.002-1.055; p = 0.0360) and in the "on" state with troublesome dyskinesia (OR, 1.032; 95% CI, 1.002-1.064; p = 0.0376) at baseline. CONCLUSIONS: Advanced PD patients with DD <10 years might benefit more from treatment with LCIG than patients with a longer DD. Although MSs severity at baseline was statistically associated with LCIG discontinuation, the probability was very low with little clinical significance.

Duodenal Levodopa Infusion for Long-Term Deep Brain Stimulation-Refractory Symptoms in Advanced Parkinson Disease.

OBJECTIVES: This study assesses the effect of levodopa/carbidopa intestinal infusion gel (LCIIG) as an additional treatment in patients with advanced idiopathic Parkinson disease (PD) previously treated with deep brain stimulation (DBS). METHODS: Prospective study of advanced PD patients, satisfactorily treated with bilateral DBS of the subthalamic nucleus, who had developed refractory symptoms and LCIIG was added. Controls were advanced PD patients treated with LCIIG. Measurements included the Unified Parkinson Disease Rating Scale (UPDRS)-III and the UPDRS axial compound. RESULTS: There were 19 patients in the DBS-LCIIG therapy group and 21 in the control group. The DBS-LCIIG patients were younger and had disease duration longer than controls. The median time from DBS to gastrostomy was 7.8 years (range, 2-12 years). In both study groups, the mean scores of the UPDRS-III and UPDRS axial subscales improved significantly after LCIIG treatment (DBS-LCIIG group: UPDRS-III, 62.0 [15.7] vs 30.9 [12.1]; UPDRS axial, 24.7 [4.9] vs 10.2 [2.7]; P < 0.0005 for all comparisons). There were no differences in adverse events between the groups. In the follow-up of the DBS-LCIIG group. 5 patients discontinued DBS-LCIIG therapy and returned to DBS, 5 discontinued DBS and were maintained with LCIIG, and the remaining 9 continued with DBS-LCIIG therapy. Mean time until discontinuation in the double DBS-LCIIG group was 891 days. The main risk factors for discontinuation were age at the beginning of LCIIG and severity of the UPDRS axial subscale. CONCLUSIONS: Levodopa/carbidopa intestinal infusion gel therapy may be a valuable option in selected patients with advanced PD who develop refractory symptoms after long-term subthalamic nucleus-DBS.

Long-term effectiveness of levodopa-carbidopa intestinal gel in 177 Spanish patients with advanced Parkinson's disease.

AIM: To assess long-term effectiveness and tolerability of levodopa-carbidopa intestinal gel (LCIG) in Spanish patients with advanced Parkinson's disease. PATIENTS & METHODS: This was an observational, multicenter, cross-sectional, retrospective study. RESULTS: Data of 177 patients were analyzed. LCIG treatment led to a reduction in the percentage of daily 'off' time (16.2 vs 47.6% before LCIG), an increase in the percentage of daily 'on' time without disabling dyskinesia (55.6 vs 21.6%). Most patients experienced improvements in freezing of gait, tremor, dizziness, fatigue or flat mood. Adverse events related to levodopa, gastrostomy and technical issues were reported in 36.2, 42.4 and 43.5% of patients, respectively. CONCLUSION: This study confirms the long-term effectiveness and safety profile of LCIG in patients with advanced Parkinson's disease.

Dissociation of eIF4E-binding protein 2 (4E-BP2) from eIF4E independent of Thr37/Thr46 phosphorylation in the ischemic stress response.

Eukaryotic initiation factor (eIF) 4E-binding proteins (4E-BPs) are translational repressors that bind specifically to eIF4E and are critical in the control of protein translation. 4E-BP2 is the predominant 4E-BP expressed in the brain, but their role is not well known. Here, we characterized four forms of 4E-BP2 detected by two-dimensional gel electrophoresis (2-DGE) in brain. The form with highest electrophoretic mobility was the main form susceptible to phosphorylation at Thr37/Thr46 sites, phosphorylation that was detected in acidic spots. Cerebral ischemia and subsequent reperfusion induced dephosphorylation and phosphorylation of 4E-BP2 at Thr37/Thr46, respectively. The induced phosphorylation was in parallel with the release of 4E-BP2 from eIF4E, although two of the phosphorylated 4E-BP2 forms were bound to eIF4E. Upon long-term reperfusion, there was a decrease in the binding of 4E-BP2 to eIF4E in cerebral cortex, demonstrated by cap binding assays and 4E-BP2-immunoprecipitation experiments. The release of 4E-BP2 from eIF4E was without changes in 4E-BP2 phosphorylation or other post-translational modification recognized by 2-DGE. These findings demonstrated specific changes in 4E-BP2/eIF4E association dependent and independent of 4E-BP2 phosphorylation. The last result supports the notion that phosphorylation may not be the uniquely regulation for the binding of 4E-BP2 to eIF4E under ischemic stress.

Neuronal apoptosis induced by cerebrospinal fluid from multiple sclerosis patients correlates with hypointense lesions on T1 magnetic resonance imaging.

Neuronal damage seems to be a major source of disability in multiple sclerosis (MS) patients and at present magnetic resonance imaging (MRI) is a sensitive method to evaluate lesion and disease activity. We studied the potential correlation between changes in MS patients' disability after relapse, the degree of T1 lesion hypointensity on MRI in vivo and neuronal apoptosis induced by cerebrospinal fluid (CSF) on neuron cultures. In this study, we included 24 MS patients with relapsing disease. Clinical recovery from relapse was measured by the Expanded Disability Status Scale (EDSS). T1-weighted MRI studies were done according to established standards and neuronal apoptosis was induced by treatment of neuronal cultures with CSF from patients while relapsing. Recovery after relapse is inversely correlated with neuronal apoptosis (r=-0.725, p<0.0001). A correlation was found between T1 lesion hypointensity and a poor recovery from relapse (r=0.656, p=0.0005) and such hypointensity correlated strongly with neuronal apoptosis (r=-0.779, p<0.0001). CSF from all patients with hypointense T1 lesions caused significantly increased neuronal apoptosis, whereas all CSF that did not induced such effects corresponded to patients without T1 lesions. The recovery from an acute MS relapse is significantly worse in patients with hypointense T1 lesions in MRI and in those whose CSF damaged neurons on cultures in vitro, phenomena that closely correlated each other.

Low concentrations of glutamate induce apoptosis in cultured neurons: implications for amyotrophic lateral sclerosis.

Evidence is accumulating that excessive glutamate concentration in the extracellular space is neurotoxic and plays a role in amyotrophic lateral sclerosis (ALS). However, the published results on glutamate levels in cerebrospinal fluid (CSF) and on glutamate-mediated toxicity of CSF in ALS disease remain controversial. In this report, we studied CSF from patients with sporadic ALS and controls to determine glutamate concentrations, and then analyzed the neurotoxic effect of glutamate at the concentrations present in CSF from ALS patients on cultured cortical neuronal cells. Our study shows that glutamate, at the concentrations found in CSF from ALS patients (5.8 microM), diminished cell viability and increased apoptosis determined by the fluorescent DNA-binding dye Hoechst 33342 as well as by Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP Nick End-Labeling (TUNEL) reaction in cultured neuronal cells. However, glutamate concentrations as those found in CSF from controls (2.8 microM or below) did not induce any effect. Both significant glutamate-induced effects were inhibited in the presence of NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline-2,3-dione), an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate-sensitive glutamate receptor antagonist. These results demonstrate that AMPA/kainate receptors are involved in the glutamate-mediated neurotoxic effects on cultured neurons, according to reports that implicate these receptors in ALS disease. We conclude that the glutamate-mediated neuronal apoptosis through AMPA/kainate receptors could occur in ALS patients who have elevated CSF glutamate concentration.

Electromyographic thresholds after thoracic screw stimulation depend on the distance of the screw from the spinal cord and not on pedicle cortex integrity.

BACKGROUND CONTEXT: Present studies concerning the safety and reliability of neurophysiological monitoring during thoracic pedicle screw placement remain inconclusive, and therefore, universally validated threshold levels that confirm osseous breakage of the instrumented pedicles have not been properly established. PURPOSE: The objective of this work was to analyze whether electromyographic (EMG) thresholds, after stimulation of the thoracic pedicle screw, depend on the distance between the neural structures and the screws. The modifier effect of different interposed tissues between a breached pedicle and neural structures was also investigated. STUDY DESIGN: This experimental study uses a domestic pig model. METHODS: Electromyographic thresholds were recorded after the stimulation of 18 thoracic pedicle screws that had been inserted into five experimental animals using varying distances between each screw and the spinal cord (8 and 2 mm). Electromyographic thresholds were also registered after the medial pedicle cortex was broken and after different biological tissues were interposed (blood, muscle, fat, and bone) between the screw and the spinal cord. RESULTS: Mean EMG thresholds increased to 14.1±5.5 mA for screws with pedicle cortex integrity that were placed 8 mm away from the dural sac. After the medial pedicle cortex was broken and without varying the distance of the screw to the dural sac, the mean EMG thresholds were not appreciably changed (13.6±6.3 mA). After repositioning the screw at a distance of 2 mm from the spinal cord and after medial cortical breach of the pedicle, the mean threshold significantly slowed to 7.4±3.4 mA (p<.001). When the screw was placed in contact with the spinal dural sac, even lower EMG thresholds were registered (4.9±1.9, p<.001). Medial pedicle cortex rupture and the interposition of different biological tissues in experimental animals did not alter the stimulation thresholds of the thoracic pedicle screws. CONCLUSIONS: In the experimental animals, the observed electrical impedance depended on the distance of screws from the neural structures and not on the integrity of the pedicle cortex. The screw-triggered EMG technique did not reliably discriminate the presence or absence of bone integrity after pedicle screw placement. The response intensity was not related to the type of interposed tissue.

Safe pedicle screw placement in thoracic scoliotic curves using t-EMG: stimulation threshold variability at concavity and convexity in apex segments.

STUDY DESIGN: A cross-sectional study of nonconsecutive cases (level III evidence). OBJECTIVE: In a series of young patients with thoracic scoliosis who were treated with pedicle screw constructs, data obtained from triggered electromyography (t-EMG) screw stimulation and postoperative computed tomographic scans were matched to find different threshold limits for the safe placement of pedicle screws at the concavity (CC) and convexity (CV) of the scoliotic curves. The influence of the distance from the medial pedicle cortex to the spinal cord on t-EMG threshold intensity was also investigated at the apex segment. SUMMARY OF BACKGROUND DATA: Whether the t-EMG stimulation threshold depends on pedicle bony integrity or on the distance to neural tissue remains elusive. Studying pedicle screws at the CC and CV at the apex segments of scoliotic curves is a good model to address this issue because the spinal cord is displaced to the CC in these patients. METHODS: A total of 23 patients who underwent posterior fusions using 358 pedicle thoracic screws were reviewed. All patients presented main thoracic scoliosis, with a mean Cobb angle of 58.3 degrees (range, 46-87 degrees). Accuracy of the screw placement was tested at surgery by the t-EMG technique. During surgery, 8 screws placed at the CC showed t-EMG threshold values below 7 mA and were carefully removed. Another 25 screws disclosed stimulation thresholds within the range of 7 to 12 mA. After checking the screw positions by intraoperative fluoroscopy, 15 screws were removed because of clear signs of malpositioning. Every patient underwent a preoperative magnetic resonance imaging examination, in which the distances from the spinal cord to the pedicles of the concave and convex sides at 3 apex vertebrae were measured. Postoperative computed tomographic scans were used in all patients to detect screw malpositioning of the final 335 screws. RESULTS: According to postoperative computed tomographic scans, 44 screws (13.1%) showed different malpositions: 40 screws (11.9%) perforated the medial pedicle wall, but only 11 screws (3.2%) were completely inside the spinal canal. If we considered the 23 screws removed during surgery, the true rate of misplaced screws increased to 18.7%. In those screws that preserved the pedicle cortex (well-positioned screws), EMG thresholds from the CC showed statistically significantly lower values than those registered at the CV of the deformity (21.1 ± 8.2 vs 23.9 ± 7.7 mA, P < 0.01). In the concave side, t-EMG threshold values under 8 mA should be unacceptable because they correspond to screw malpositioning. Threshold values above 14 mA indicate an accurate intrapedicular position with certainty. At the convex side, threshold values below 11 mA always indicate screw malpositioning, and values above 19 mA imply accurate screw placement. At the 3 apex vertebrae, the average pedicle-spinal cord distance was 2.2 ± 0.7 mm at the concave side and 9.8 ± 4.3 mm at the convex side (P < 0.001). In well-positioned screws, a correlation between pedicle-dural sac distance and t-EMG threshold values was found at the concave side only (Pearson r = 0.467, P < 0.05). None of the patients with misplaced screws showed postoperative neurological impairment. CONCLUSION: Independent of the screw position, average t-EMG thresholds were always higher at the CV in the apex and above the apex regions, presuming that the distance from the pedicle to the spinal cord plays an important role in electrical transmission. The t-EMG technique has low sensitivity to predict screw malpositioning and cannot discriminate between medial cortex breakages and complete invasion of the spinal canal.