Surgical Outcome in Extratemporal Epilepsies Based on Multimodal Pre-Surgical Evaluation and Sequential Intraoperative Electrocorticography.

OBJECTIVE: to present the postsurgical outcome of extratemporal epilepsy (ExTLE) patients submitted to preoperative multimodal evaluation and intraoperative sequential electrocorticography (ECoG). SUBJECTS AND METHODS: thirty-four pharmaco-resistant patients with lesional and non-lesional ExTLE underwent comprehensive pre-surgical evaluation including multimodal neuroimaging such as ictal and interictal perfusion single photon emission computed tomography (SPECT) scans, subtraction of ictal and interictal SPECT co-registered with magnetic resonance imaging (SISCOM) and electroencephalography (EEG) source imaging (ESI) of ictal epileptic activity. Surgical procedures were tailored by sequential intraoperative ECoG, and absolute spike frequency (ASF) was calculated in the pre- and post-resection ECoG. Postoperative clinical outcome assessment for each patient was carried out one year after surgery using Engel scores. RESULTS: frontal and occipital resection were the most common surgical techniques applied. In addition, surgical resection encroaching upon eloquent cortex was accomplished in 41% of the ExTLE patients. Pre-surgical magnetic resonance imaging (MRI) did not indicate a distinct lesion in 47% of the cases. In the latter number of subjects, SISCOM and ESI of ictal epileptic activity made it possible to estimate the epileptogenic zone. After one- year follow up, 55.8% of the patients was categorized as Engel class I-II. In this study, there was no difference in the clinical outcome between lesional and non lesional ExTLE patients. About 43.7% of patients without lesion were also seizure- free, p = 0.15 (Fischer exact test). Patients with satisfactory seizure outcome showed lower absolute spike frequency in the pre-resection intraoperative ECoG than those with unsatisfactory seizure outcome, (Mann- Whitney U test, p = 0.005). CONCLUSIONS: this study has shown that multimodal pre-surgical evaluation based, particularly, on data from SISCOM and ESI alongside sequential intraoperative ECoG, allow seizure control to be achieved in patients with pharmacoresistant ExTLE epilepsy.

New Aspects of VEGF, GABA, and Glutamate Signaling in the Neocortex of Human Temporal Lobe Pharmacoresistant Epilepsy Revealed by RT-qPCR Arrays.

In the epilepsy spectrum, temporal lobe epilepsy (TLE) is the most common and devastating focal and symptomatic epilepsy form in adults, where more than 30% of patients develop pharmacoresistance. It is not fully understood how the gene expression contributes to establishing an epileptic phenotype. Cerebrovascular remodeling directed by VEGF (vascular endothelial growth factor) signaling might modulate the synaptic neurotransmission in the epileptic brain. To address this question, the gene expression was profiled in biopsies of the temporal cortex from diagnosed patients with pharmacoresistant TLE that underwent surgical resection to seizure control. One hundred sixty-eight genes related to VEGF signaling and GABA and glutamate neurotransmissions were evaluated. Genes related to downstream signaling -phosphoinositide 3-kinase (PI3K), mitogen-activated protein kinases (MAPK), and Janus-activated kinase/signal transducer and activator of transcription (JAK/STAT) pathways- and neurotransmitters metabolism were evaluated too. Thirty-nine genes were upregulated. The genes encoding for G protein q polypeptide, serine racemase, gephyrin, and glutamate/cystine antiporter system xCT appeared as novel upregulated genes in the pharmacoresistant TLE. ClueGO, a Cytoscape plugin, was used to build a gene network associated using Gene Ontology (GO) terminology. Enrichment analysis by ClueGO retrieves that positive regulation of endothelial cell proliferation, nerve development, and neuronal apoptosis were over-represented categories. In conclusion, VEGF signaling is confirmed as a relevant mediator in the pharmacoresistant TLE. In addition, the enrichment analysis applied to differentially expressed genes suggests new pharmacological targets to be assessed in the treatment of pharmacoresistant TLE. Results make up an approximation to better understand the epileptic brain and complement the available data.

Relation of Structural and Functional Changes in Auditory and Visual Pathways after Temporal Lobe Epilepsy Surgery.

Auditory and visual pathways may be affected as a consequence of temporal lobe epilepsy surgery because of their anatomical relationships with this structure. The purpose of this paper is to correlate the results of the auditory and visual evoked responses with the parameters of tractography of the visual pathway, and with the state of connectivity between respective thalamic nuclei and primary cortices in both systems after the surgical resection of the epileptogenic zone in drug-resistant epileptic patients. Tractography of visual pathway and anatomical connectivity of auditory and visual thalamus-cortical radiations were evaluated in a sample of eight patients. In general, there was a positive relationship of middle latency response (MLR) latency and length of resection, while a negative correlation was found between MLR latency and the anatomical connection strength and anatomical connection probability of the auditory radiations. In the visual pathway, significant differences between sides were found with respect to the number and length of tracts, which was lower in the operated one. Anatomical connectivity variables and perimetry (visual field defect index) were particularly correlated with the latency of P100 wave which was obtained by quadrant stimulation. These results demonstrate an indirect functional modification of the auditory pathway and a direct traumatic lesion of the visual pathway after anterior temporal lobectomy in patients with drug resistant epilepsy.

Rotating and Neurochemical Activity of Rats Lesioned with Quinolinic Acid and Transplanted with Bone Marrow Mononuclear Cells.

Huntington's disease (HD) is an inherited, neurodegenerative disorder that results from the degeneration of striatal neurons, mainly GABAergic neurons. The study of neurochemical activity has provided reliable markers to explain motor disorders. To treat neurodegenerative diseases, stem cell transplants with bone marrow (BM) have been performed for several decades. In this work we determine the effect of mononuclear bone marrow cell (mBMC) transplantation on the rotational behavior and neurochemical activity in a model of Huntington's disease in rats. Four experimental groups were organized: Group I: Control animals (n = 5); Group II: Lesion with quinolinic acid (QA) in the striatum (n = 5); Group III: Lesion with QA and transplant with mBMC (n = 5); Group IV: Lesion with QA and transplant with culture medium (Dulbecco's modified Eagle's medium (DMEM) injection) (n = 5). The rotational activity induced by D-amphetamine was evaluated and the concentration of the neurotransmitter amino acids (glutamate and GABA) was studied. The striatal cell transplantation decreases the rotations induced by D-amphetamine (p < 0.04, Wilcoxon matched pairs test) and improves the changes produced in the levels of neurotransmitters studied. This work suggests that the loss of GABAergic neurons in the brain of rats lesioned with AQ produces behavioral and neurochemical alterations that can be reversed with the use of bone marrow mononuclear cell transplants.

Follow-Up of Peripheral IL-1ß and IL-6 and Relation with Apoptotic Death in Drug-Resistant Temporal Lobe Epilepsy Patients Submitted to Surgery.

Increasing amounts of evidence support the role of inflammation in epilepsy. This study was done to evaluate serum follow-up of IL-1ß and IL-6 levels, as well as their concentration in the neocortex, and the relationship of central inflammation with NF-κB and annexin V in drug-resistant temporal lobe epileptic (DRTLE) patients submitted to surgical treatment. Peripheral and central levels of IL-1ß and IL-6were measured by ELISA in 10 DRTLE patients. The sera from patients were taken before surgery, and 12 and 24 months after surgical treatment. The neocortical expression of NF-κB was evaluated by western blotting and annexin V co-localization with synaptophysin by immunohistochemistry. The neocortical tissues from five patients who died by non-neurological causes were used as control. Decreased serum levels of IL-1 and IL-6 were observed after surgery; at this time, 70% of patients were seizure-free. No values of IL-1 and IL-6 were detected in neocortical control tissue, whereas cytokine levels were evidenced in DRTLE. Increased NF-κB neocortex expression was found and the positive annexin V neurons were more obvious in the DRTLE tissue, correlating with IL-6 levels. The follow-up study confirmed that the inflammatory alterations disappeared one year after surgery, when the majority of patients were seizure-free, and the apoptotic death process correlated with inflammation.

Oxidative Stress in Patients with Drug Resistant Partial Complex Seizure.

Oxidative stress (OS) has been implicated as a pathophysiological mechanism of drug-resistant epilepsy, but little is known about the relationship between OS markers and clinical parameters, such as the number of drugs, age onset of seizure and frequency of seizures per month. The current study’s aim was to evaluate several oxidative stress markers and antioxidants in 18 drug-resistant partial complex seizure (DRPCS) patients compared to a control group (age and sex matched), and the results were related to clinical variables. We examined malondialdehyde (MDA), advanced oxidation protein products (AOPP), advanced glycation end products (AGEs), nitric oxide (NO), uric acid, superoxide dismutase (SOD), glutathione, vitamin C, 4-hydroxy-2-nonenal (4-HNE) and nitrotyrosine (3-NT). All markers except 4-HNE and 3-NT were studied by spectrophotometry. The expressions of 4-HNE and 3-NT were evaluated by Western blot analysis. MDA levels in patients were significantly increased (p ≤ 0.0001) while AOPP levels were similar to the control group. AGEs, NO and uric acid concentrations were significantly decreased (p ≤ 0.004, p ≤ 0.005, p ≤ 0.0001, respectively). Expressions of 3-NT and 4-HNE were increased (p ≤ 0.005) similarly to SOD activity (p = 0.0001), whereas vitamin C was considerably diminished (p = 0.0001). Glutathione levels were similar to the control group. There was a positive correlation between NO and MDA with the number of drugs. The expression of 3-NT was positively related with the frequency of seizures per month. There was a negative relationship between MDA and age at onset of seizures, as well as vitamin C with seizure frequency/month. We detected an imbalance in the redox state in patients with DRCPS, supporting oxidative stress as a relevant mechanism in this pathology. Thus, it is apparent that some oxidant and antioxidant parameters are closely linked with clinical variables.

Long-Term Electroclinical and Employment Follow up in Temporal Lobe Epilepsy Surgery. A Cuban Comprehensive Epilepsy Surgery Program.

The purpose of this paper is to present a long- term electroclinical and employment follow up in temporal lobe epilepsy (TLE) patients in a comprehensive epilepsy surgery program. Forty adult patients with pharmacoresistant TLE underwent detailed presurgical evaluation. Electroencephalogram (EEG) and clinical follow up assessment for each patient were carried out. The occurrence of interictal epileptiform activity (IEA) and absolute spike frequency (ASF) were tabulated before and after 1, 6, 12, 24 and 72 months surgical treatment. Employment status pre- to post-surgery at the last evaluated period was also examined. Engel scores follow-up was described as follows: at 12 months 70% (28) class I, 10% (4) class II and 19% (8) class III-IV; at 24 months after surgery 55.2% (21) of the patients were class I, 28.9% (11) class II and 15.1% (6) class III-IV. After one- year follow up 23 (57.7%) patients were seizure and aura-free (Engel class IA). These figures changed to 47.3%, and 48.6% respectively two and five years following surgery whereas 50% maintained this condition in the last follow up period. A decline in the ASF was observed from the first year until the sixth year after surgery in relation to the preoperative EEG. The ASF one year after surgery allowed to distinguish "satisfactory" from "unsatisfactory" seizure relief outcome at the last follow up. An adequate social functioning in terms of education and employment in more than 50% of the patients was also found. Results revealed the feasibility of conducting a successful epilepsy surgery program with favorable long term electroclinical and psychosocial functioning outcomes in a developing country as well.

Autoradiography reveals selective changes in serotonin binding in neocortex of patients with temporal lobe epilepsy.

The main goal of the present study was to evaluate binding to serotonin in the neocortex surrounding the epileptic focus of patients with mesial temporal lobe epilepsy (MTLE). Binding to 5-HT, 5-HT(1A), 5-HT(4), 5-HT(7) receptors and serotonin transporter (5-HTT) in T1-T2 gyri of 15 patients with MTLE and their correlations with clinical data, neuronal count and volume were determined. Autopsy material acquired from subjects without epilepsy (n=6) was used as control. The neocortex from MTLE patients demonstrated decreased cell count in layers III-IV (21%). No significant changes were detected on the neuronal volume. Autoradiography experiments showed the following results: reduced 5-HT and 5-HT(1A) binding in layers I-II (24% and 92%, respectively); enhanced 5-HT(4) binding in layers V-VI (32%); no significant changes in 5-HT(7) binding; reduced 5-HTT binding in all layers (I-II, 90.3%; III-IV, 90.3%, V-VI, 86.9%). Significant correlations were found between binding to 5-HT(4) and 5-HT(7) receptors and age of seizure onset, duration of epilepsy and duration of antiepileptic treatment. The present results support an impaired serotoninergic transmission in the neocortex surrounding the epileptic focus of patients with MTLE, a situation that could be involved in the initiation and propagation of seizure activity.

Transient glutathione depletion in the substantia nigra compacta is associated with neuroinflammation in rats.

Glutathione (GSH) deficiency has been identified as an early event in the progression of Parkinson's disease. However, the role of GSH in the etiology and pathogenesis of this neurodegenerative disorder is not well established. The aim of this study is to assess the effect of transient GSH depletion in the substantia nigra pars compacta (SNpc) on neuroinflammation after the injection of a single dose of l-buthionine sulfoximine (BSO) into the SNpc of male Sprague-Dawley rats. The results showed that BSO treatment stimulates microglia (p<0.01) and astroglial response (p<0.01), c-Jun N-terminal kinase and inducible nitric oxide synthase (iNOS) (p<0.001) in the SNpc, accompanied by dopaminergic dysfunction. In addition, high levels of tumor necrosis factor α (p<0.01), interleukins IL-1ß p<0.01), IL-6 p<0.001) and nitric oxide p<0.01) were found in the treated animals compared to control groups, while no significant differences were found in IL-10 levels. These results suggest that transient GSH depletion can increase the susceptibility of SNpc to degeneration by promoting an inflammatory response and nitrosative stress, reinforcing the possible role of GSH unbalance, oxygen/nitrogen reactive species and neuroinflammation as causal factors on the degeneration of the SNpc.

Multimodal imaging in nonlesional medically intractable focal epilepsy.

Identification and localization of epileptogenic zone (EZ) is vital in patients with medically-intractable focal epilepsy, who may be candidates for potentially curative resective epilepsy surgery. Presence of a lesion on magnetic resonance imaging (MRI) influences both diagnostic classification and selection for surgery. However, the implications for MRI-negative cases are not well-defined for such patients. Most of these patients undergo invasive long-term Electroencephalography recordings before a final decision regarding resection is possible. Recent developments in structural and functional neuroimaging which include quali-quantitative MRI, Positron Emission Tomography, Single Photon Emission Computed Tomography, and functional MRI have significantly changed presurgical epilepsy evaluation. Source analysis based on electrophysiological information, using either EEG or magnetoencephalography are also promising in order to noninvasively localize the EZ and to guide surgery in medically-intractable focal epilepsy patients that exhibit nonlesional MRI. This chapter aims to review the value of the combined use of structural and functional imaging techniques, and how this multimodal approach improves both selection of surgical candidates and post-operative outcomes in medically-intractable nonlesional focal epilepsy.

Nerve growth factor levels in Parkinson disease and experimental parkinsonian rats.

Nerve growth factor (NGF) is well established for its ability to promote growth and survival for specific neuronal populations. However, its participation in the pathogenesis of human nervous system disorders such as Parkinson's disease (PD) remains to be resolved. This study examined NGF levels in the serum of healthy persons, in patients with PD and in parkinsonian rats using a double site immune-enzymatic assay (EIA) with the murine 27/21 anti-beta-NGF monoclonal antibody. PD patients were divided in two groups according to the stages of the disease (Grade: I-II and Grade: III-IV of Hoenh and Yahr scale). NGF levels in parkinsonian rats showed significant (P<0.01) reductions when compared with serum from normal animals. The NGF levels in early states of the disease (Grade I-II) showed greater reductions (P<0.01) in comparison to those with advanced stages (Grade III-IV). We consider that alterations in NGF levels may reflect ongoing neurodegenerative processes in PD.

Temporal lobe epilepsy surgery modulates the activity of auditory pathway.

The purpose of this paper is to evaluate the effects of the anterior temporal lobectomy on the functional state of the auditory pathway in a group of drug-resistant epileptic patients, linking the electrophysiological results to the resection magnitude. Twenty-seven patients with temporal lobe epilepsy and a matched control group were studied. Auditory brainstem and middle latency responses (ABR and MLR respectively) were carried out before and after 6, 12 and 24 months surgical treatment. The volume and longitude of temporo-mesial resected structures were estimated on magnetic resonance images taken 6 months after surgery. Before the intervention the patients showed a significant delay of latency in waves III, V, Pa and Nb, with an increase in duration of I-V interval in comparison with healthy subjects (Mann-Whitney U-test, p<0.05). After resection, additional significant differences in waves I and Na latency were observed. Na and Pa waveforms showed a tendency to increase in amplitude, which became statistically significant 12 months after surgery for right hemisphere lobectomized patients in the midline electrode, and in Pa waveform for all patients in the temporal electrodes ipsilateral to resection (Wilcoxon test, p<0.05). In general, latency variations of MLR correlated with resection longitude, while changes in amplitude correlated with the volume of the resection in the middle temporal pole and amygdala (Pearson' correlation test, p<0.05). As a result, we assume that anterior temporal lobectomy provokes functional modifications into the auditory pathway, probably related to an indirect modulation of its activity by the temporo-mesial removed structures.

Inflammatory mediators in epilepsy.

All common contributing factors to epilepsy such as trauma, malignancies and infections are accompanied by different levels of central nervous system inflammation that in turn have been associated with the occurrence of seizure. Emerging data from human brain tissue and experimental models of epilepsy support the proposed involvement of inflammation in epilepsy. Key mediators of this process include, among others: interleukin (IL) -1ß, IL-6, tumor necrosis factor-α, adhesion molecules and component of complement. Recent advances suggest the involvement of specific inflammatory pathways in the pathogenesis of seizures in patients with pharmacoresistant temporal lobe epilepsy, highlighting the potential for new therapeutic strategies. This review provides an overview of the current knowledge on the relationship between inflammatory mediators and epilepsy. We also describe experimental and clinical evidence of inflammation in epilepsy with special emphasis on clinical aspects once the epileptogenic focus has been resected. Further insight into the complex role of inflammation in epileptogenesis may provide new treatment options.

Mecanismos de neurodegeneración en la epilepsia del lóbulo temporal / Mechanisms of neurodegeneration in temporal lobe epilepsy

Epilepsy affects 1 and 2 percent of the worldwide population, while temporal lobe epilepsy (TLE) covers 40 percent of all epilepsy cases. Controversy in defining epilepsy as a neurodegenerative disease exists because, no there is enough evidence to show seizures and status epilepticus (SE) as cause for irreversible neuronal damage. Epileptogenic insult at the beginning of the disease produces an acute and delayed neuronal death, resulting in gliosis, but also triggers compensatory processes such as angiogenesis, cell proliferation and reorganization of extracellular matrix as receptors, channels and drug transporter proteins. In neurogenesis and axonal regrowth, the age of onset is crucial for the formation of abnormal neurons and aberrant circuits as a result of seizures; approximately 30 percent begin in the temporal lobe. These disturbances continue in parallel or sequentially during the course of epilepsy, which implies a great challenge in the search of new treatments...

La epilepsia es una enfermedad que afecta entre el 1 al 2 por ciento de la población mundial, siendo la epilepsia del lóbulo temporal (ELT) la que abarca el 40 por ciento de todos los casos de epilepsia. La controversia en definir a la epilepsia como una enfermedad neurodegenerativa, se debe a que no hay pruebas suficientes que demuestren como las convulsiones y el estado de mal epiléptico (SE) provocan un daño neuronal irreversible. El insulto epileptógenico presente al inicio de la enfermedad genera la muerte neuronal aguda y tardía, para dar lugar a la gliosis; pero también se desencadenan procesos compensatorios como la angiogénesis, la proliferación celular y una reorganización tanto de la matriz extracelular como de los receptores, canales y proteínas transportadoras de fármacos. En el caso de la neurogénesis y recrecimiento axonal, la edad de inicio es determinante para la formación de neuronas anormales y circuitos aberrantes como consecuencia de las convulsiones, dónde aproximadamente un 30 por ciento comienzan en el lóbulo temporal. Estas alteraciones se continúan en paralelo o de forma secuencia! durante la evolución de la epilepsia, lo que implica un gran desafío en la búsqueda de nuevos tratamientos...

El núcleo pedunuulopontino y su relación con la fisiopatología de la enfermedad de parkinson / The Pedunculopontine Nucleus and its Relationship to the Pathophysiology of Parkinson's Disease

Varias décadas de investigaciones neuropatológicas e imagenológicas han proporcionado suficientes evidencias acerca de las alteraciones en la neurotransmisión colinérgica que acompañan a la disfunción dopaminérgica en la enfermedad de Parkinson (EP). El núcleo pedunculopontino tegmental laterodorsal (NPP) representa una de las fuentes principales de proyecciones colinérgicas en el cerebro y a su vez es el origen de la única proyección colinérgica que recibe la substantia nigra pars compacta (SNpc). Actualmente el estudio de la participación del NPP en la fisiopatología de la EP toma en cuenta dos vertientes: el impacto de la pérdida temprana de la influencia excitatoria pontina sobre la SNpc asociado a la degeneración temprana del NPP y la estimulación a baja frecuencia del NPP como tratamiento quirúrgico beneficioso para los signos axiales de la EP. El NPP ha emergido como una estructura esencial en la comprensión de la fisiopatología de la EP dado sus relaciones con los núcleos de los ganglios basales, el tálamo, la corteza motora y la médula espinal. La degeneración de algunas de sus poblaciones neuronales en etapas presintomáticas de la EP ha sugerido una relación causa-efecto entre este hallazgo y la muerte de las células dopaminérgicas nigrales. Por otra parte la estimulación del NPP tiene resultados favorables sobre los trastornos posturales y de la marcha, los cuales se presentan en etapas tardías de la EP y son refractarios a otros tratamientos farmacológicos y quirúrgicos.

Several decades of neuropathologic and imagenologic investigations have provided sufficient evidences about alterations in cholinergic neurotransmission that go together with the dopaminergic dysfunction in Parkinson s disease (PD). The laterodorsal tegmental pedunculopontine nucleus (PPN) represents one of the main sources of cholinergic projections into the brain and at the same time the origin of the only cholinergic projection that substantia nigra pars compacta (SNpc) receives. At present, the study of the PPN participation as part of the physiopathology of PD has two notions: the impact of the lack of pontine excitatory influence on SNpc, associated to the early degeneration of PPN as well as the low frequency stimulation in the PPN as a beneficial surgical treatment for the axial symptoms of PD. PPN has emerged as an essential structure in the comprehension of PD physiopathology, given by its relation with the basal ganglia nuclei, thalamus, motor cortex and the spinal cord. The degeneration of some of its neuronal populations in PD pre symptomatic steps, has suggested a cause- and-effect relation on this finding and the death of nigral dopaminergic cells. On the other hand, PPN stimulation has favorable results on postural and gait disorders, which present themselves in late PD stages and are refractory to other pharmacological and surgical treatments.

Efecto de la adiinistración intracerebral de mk801 y (-) nicotina en las conceentraciones extracelulares de glu y gaba en el núcleo pedunculopontino de ratas / Effect of the Mk801 and (-) Nicotine Intracerebral Administration on Glu and Gaba Extracellular Concentration in the Pedunculopontine Nucleus from Rats

Aunque la manipulación farmacológica de los sistemas glutamatérgico y colinérgico se ha tratado en modelos experimentales de enfermedad de Parkinson (EP), pocos autores han realizado estudios de esta temática a nivel del núcleo pedunculopontino (NPP). El presente trabajo aborda los cambios en las concentraciones extracelulares (CE) de glutamato (Glu) y ácido δ-amino butírico (GABA) en el NPP de ratas hemiparkinsonizadas por inyección de 6-hidroxidopamina (6-OHDA) y sometidas a infusión local de MK-801 (10 µmol/L) o (-) nicotina (10 mM). La infusión se realizó mediante microdiálisis cerebral y la determinación de CE de neurotransmisores se realizó a través de cromatografía líquida de alta resolución acoplada a detección de fluorescencia. La infusión de MK-801 en el NPP produjo disminución significativa de CE de Glu (p< 0,01) y de GABA (p < 0,01) en ratas hemiparkinsonizadas y controles. La infusión de (-) nicotina mostró un incremento significativo de CE de Glu (p < 0,001) y GABA (p< 0,001) en el NPP de ratas hemiparkinsonizadas y controles. El bloqueo local de receptores NMDA por MK-801 facilita la interacción de Glu con sus receptores metabotrópicos que participan en mecanismos de inhibición presináptica y bloquean la liberación de neurotransmisores. Mientras que la infusión de nicotina en el NPP suma los efectos de activación de los receptores nicotínicos a los cambios conocidos en la neurotransmisión glutamatérgica y gabaérgica en el NPP en parkinsonismo. La infusión de fármacos glutamatérgicos y colinérgicos en el NPP, impone un reajuste a la neurotransmisión a este nivel que se añade a los cambios neuroquímicos asociados a denervación dopaminérgica.

Although the pharmacological manipulation of the glutamatergic and cholinergic systems have been studied in animal models of Parkinson´s disease (PD), only some authors have done work on this topic at the pedunculopontine nucleus (PPN). The present work studied the changes in glutamate (Glu) and δ-aminobutyric acid (GABA) extracellular concentrations (EC) in the PPN from hemiparkinsonian rats by 6hydroxydopamine injection. The rats were locally perfused by MK-801 (10 µmol/L) or (-) nicotine (10 mM) solutions by cerebral microdyalisis. The biochemical studies were carried out through high performance liquid chromatography coupled to fluorescence detection. MK-801 infusion induced a significant decrease of Glu (p< 0.01) and GABA (p< 0.01) EC in PPN. On the other hand (-) nicotine infusion induced a significant increase of Glu (p< 0.001) and GABA (p< 0.001) EC in PPN from hemiparkinsonian rats. The local blockade of NMDA receptors by MK-801 infusion facilitates the interaction between Glu and their metabotropic receptors that take part in presynaptic inhibition mechanisms and interfere with neurotransmitters release. Meanwhile, the nicotine infusion sums the effects of nicotinic receptor activation with the glutamatergic and gabaergic neurotransmission changes produced in the PPN in the parkinsonian condition. The cholinergic and glutamergic drug infusion in PPN impose a new adjustment to the neurotransmition at this level that is added to the neurochemical changes associated to dopaminergic denervation.

Establecimiento de las condiciones para el trasplante de células de médula ósea en un modelo de enfermedad de Huntington y su efecto funcional a través de la conducta motora / Establishment of the conditions for the bone marrow cells transplants in a model of Huntington´s disease and functional effect on the motor behavior

La enfermedad de Huntington (EH) es un trastorno degenerativo hereditario que afecta a personas con predisposición genética. No existe hasta hoy un tratamiento efectivo; la enfermedad avanza lentamente y el paciente termina en incapacidad o muerte después de 15 o 20 años. Los estudios relacionados con el tratamiento de las manifestaciones clínicas que aparecen en la enfermedad, incluyen tratamientos medicamentosos y el uso de trasplante de células. En la actualidad se conoce que es posible reproducir algunas características de la enfermedad en modelos experimentales para ensayar posibles terapéuticas (ej. el modelo de lesión estriatal por inyección de ácido quinolínico; [AQ]). No se conoce el efecto restaurativo de las células de médula ósea (CMO) en este modelo. Objetivos: 1) Caracterizar morfológicamente la lesión por inyección intraestriatal de AQ. 2) Caracterizar inmunocitoquímicamente las CMO. 3) Evaluar la concentración óptima de CMO para el trasplante en el modelo y 4) Evaluar el estado funcional del trasplante de CMO, a través de la conducta motora.

Huntington Disease (HD) is a heritable neurodegenerative disease that affects people with genetic history. Until today, an effective treatment doesn't exist; the illness advances slowly and the patient finishes in inability or death after 15 or 20 years. The studies related with the treatment of the clinical manifestations, include treatments with medications and the use of cells transplant. At the present time it is known that it is possible to reproduce, some characteristics of the disease in experimental models for to use possible therapies [example: estriatal lesion of quinolínico acid; (QA)]. the restorative effect of the bone marrow cells (BMC) is not known in this model. Objectives. 1) characterizationmorphofological of the estriatal lesion whith QA. 2) to characterization immunochemical of BMC. 3) to evaluate the BMC concentration for the transplant and 4) to evaluate the functional state of BMC transplant, through the motor behavior.

Enfermedad de hnntington: modelos experimentales y perspectivas terapéuticas / Huntington'disease: Experimentals Models and Therapeutic Perspectives

La enfermedad de Huntington (EH) es un trastorno degenerativo de Weiss de origen hereditario. Hasta el momento no existe un tratamiento efectivo para la enfermedad que inexorablemente después de transcurridos 15 a 20 años, evoluciona hacia incapacidad total o muerte. En este trabajo se revisan las características clínicas y morfológicas de la EH y los modelos experimentales más utilizados para su estudio tomando como fuente, artículos indexados en la base de datos Medline publicados en los últimos 20 años. Se valoran las ventajas y desventajas de estos modelos y su perspectiva para el desarrollo de ensayos clínicos. El consenso de lo reportado plantea que de los modelos tóxicos, los inducidos por neurotoxinas tales como ácido quinolínico parecen ser los más adecuados para reproducir las características neuropatológicas, y por otro lado los modelos genéticos contribuyen con más evidencias al conocimiento del origen etiológico de la enfermedad. Numerosos tratamientos han sido aplicados en el manejo de las manifestaciones clínicas que aparecen en EH, sin poder detener o disminuir las afectaciones que derivan de la pérdida neuronal. La sintomatología clínica ha sido posible reproducirla, al menos en parte, en animales de experimentación lo que ha permitido realizar ensayos terapéuticos. Desde el punto de vista de tratamiento, lo que más promisorio parece ser, la terapia celular con células provenientes de diferentes fuentes y dentro de ellas las no neurales, que implican menor censura ética y mayor factibilidad de obtención para la aplicación en los enfermos. Por otro lado el desarrollo de la tecnología del ARN de interferencia, emerge como una herramienta terapéutica potencial para el tratamiento de EH, así como para responder interrogantes básicas relacionadas con el desarrollo de la enfermedad.

Huntington'disease (HD) is a degenerative dysfunction of hereditary origin. Up to date there is not, an effective treatment to the disease which having lapsed 15 or 20 years advances inexorably, in a slow form, toward the total inability or death. This paper reviews the clinical and morphological characteristics of Huntington's disease as well as the experimental models more commonly used t study this disease, having as source the articles indexed in Medline data base, published in the last 20 years. Advantages and disadvantages of all experimental models to reproduce the disease as well as the perspectives to therapeutic assay have been also considered. The consent of outline reported about the toxic models, those induced by neurotoxins such as quinolinic acid, appears to be the most appropiate to reproduce the neuropathologic characteristic of the disease, an genetic models contributing with more evidence to the knowledge of the disease ethiology. Numerous treatments ameliorate clinical manifestations, but none of them has been able to stop or diminish the affectations derived from neuronal loss. At present time it is possible to reproduce, at least partially, the characteristics of the disease in experimentation animals that allow therapy evaluation in HD. From the treatment view point, the more promissory seems to be transplantation of no neuronal cells, taking into account ethical issues and factibility. On the other hand the new technology of interference RNA, emerges as a potential therapeutic tool for treatment in HD, and to respond basic questions on the development of the disease.

Aplicación del eest de la barra transversal modficado para evaluar ratas hemipparkinsonizadas. / Modify Beam Transversal Test to Evauate Hemiparkinsonian Rats.

La degeneración nigroestriatal que caracteriza a la enfermedad de Parkinson (EP) es estudiada en modelos experimentales en roedores por inyección de 6-hidroxidopamina (6-OHDA). El presente estudio presenta una versión modificada del test de la barra transversal (TBT) que permite la cuantificación del déficit motor a través de: tiempo que demora la rata en alcanzar una de las plataformas (latencia de escape, LE); tiempo que demora en caer de la barra (latencia de caída, LC); número total de errores cometidos durante la ejecución en cada barra (número de errores, NE). La forma y el diámetro de la sección transversal de la barra se modificaron desde barras rectangulares y circulares de 2,5 cm de diámetro hasta barras con esta misma forma y 1 cm de diámetro respectivamente lo cual impuso la mayor dificultad a la ejecución del test. Tres grupos de ratas Wistar fueron evaluados: no tratadas (n=15), lesionadas con 6-OHDA (n=14) y falsas operadas (n=14). Todas las variables estudiadas mostraron diferencias signifi-cativas entre ratas controles y hemiparkinsonizadas. Para todos los tipos de barras, las variables LE y NE se incrementaron mientras que la LC disminuyó significativamente en las ratas hemiparkinsonizadas en comparación con las ratas controles. La LC mostró diferencias altamente significativas (p<0,001) entre las barras de mayor y menor diámetro. TBT es un test que explora la función sensoriomotora, no requiere grandes sesiones de entrenamiento previo ni motivación aversiva ni deprivación de alimento. Este test resulta de gran utilidad para evaluar las deficiencias motoras que se presentan en el modelo de hemiparkinsonismo unilateral así como en otros modelos experimentales de enfermedades neurodegenerativas.

The nigrostriatal degeneration underlying Parkinson’s disease (PD) is commonly studied in experimental animals by injection of the neurotoxin 6-hydroxydopamine. The present study describes a modified version of a beam traversal test which allows the quantification of the motor deficit through the time spent to arrive to the platform once all four paws of the animals are in contact with the beam (escape latency, EL), the time spent before falling (tumbled down latency, TDL) and the number of errors (NE) committed for the animals in each beam. The shape and the diameter of the cross section of the beams were modified from rectangular and circular cross section with 2,5 cm of diameter to the same shape with 1 cm of diameter, which induced a high difficulty to the execution of the test. Three groups of Wistar rats were examined: untreated (n=15), lesioned with 6-hydroxydopamine (n=14), and sham-operated (n=14). All variables studied showed significant differences between control and hemiparkinsonian rats. The EL and the NE were increased and the TDL was decreased in hemiparkinsonian rats for all beams in comparison with control rats. In TDL the significant differences between groups were more evident (p<0.001) for the beams with high cross section irrespective of the shape of the cross section. BTT is a convenient sensorimotor test that does not need to be trained extensively, and require adverse motivation or food deprivation and appears to be very useful in evaluating the motor deficits in established unilateral model of PD and also other experimental models.

Estudio de la función citotóxica de las células K y las subpoblaciones de células T en pacientes con anemia aplástica (AA) / Study of cytotoxic function of K cells and T cell subpopulations in patients with aplastic anemia (AA)

En 17 pacientes con anemia aplástica se estudiaron con sangre periférica: la función de las células citotóxicas (K) y los linfocitos T (CD3 a la + y CD2 a la +), la subpoblación auxiliadora-inductora (CD4 a la +) y supresora-citotóxica (CD8 a la +). Se compararon los resultados con los obtenidos en un grupo de controles normales. Se encontró una disminución estadísticamente significativa en la función citotóxica de las células K y en las subpoblaciones de células CD4 a la + y CD8 a la +, así como el índice CD4/CD8