Biopsia cerebral estereotáctica sin marco: capacidad diagnóstica y complicaciones / Frameless stereotactic biopsy: Diagnostic yield and complications

OBJETIVO: El objetivo de este estudio fue evaluar las variables que condicionan el rendimiento diagnóstico de la BSTsm y las complicaciones de esta técnica. MATERIAL Y MÉTODO: Estudio retrospectivo de las BSTsm realizadas entre julio del 2008 y diciembre de 2011 en el Hospital Universitario Donostia. En el que estudiamos las variables tamaño, distancia al córtex, captación de contraste y localización. RESULTADOS: Incluimos 70 pacientes (75 biopsias), 39 varones y 31 mujeres con un rango de edad entre 39 y 83 años. El rendimiento diagnóstico total de nuestra serie fue del 97,1%. El punto de corte de la variable tamaño, a partir del cual encontramos el rendimiento más alto fue de 19 mm; así para las lesiones > 19 mm se obtuvo una sensibilidad del 95,2% (IC 95%: 86,9-98,4) y una especificidad del 57,1% (IC 95%: 25,0-84,2). Las lesiones localizadas a menos de 17 mm del córtex mostraron un rendimiento menor, con una sensibilidad del 74,6% (IC 95%: 62,1-84,7) y una especificidad del 71,4% (IC 95%: 29,0-96,3). Siete (10%) pacientes desarrollaron complicaciones después de la primera biopsia y ninguno tras la segunda. CONCLUSIONES: El rendimiento diagnóstico fue menor en las lesiones menores de 2 cm de diámetro y superficiales (< 17 mm del córtex). En esta serie no observamos un aumento de las complicaciones después de realizar una segunda biopsia

OBJECTIVE: The aim of this study was to evaluate the variables that could modify the diagnostic yield of frameless stereotactic biopsy, as well as its complications. MATERIALS AND METHOD: This was a retrospective study of frameless stereotactic biopsies carried out between July 2008 and December 2011 at Donostia University Hospital. The variables studied were size, distance to the cortex, contrast uptake and location. RESULTS: A total of 70 patients were included (75 biopsies); 39 males and 31 females with an age range between 39 and 83 years. The total diagnostic yield in our series was 97.1%. For lesions > 19 mm, the technique offered a sensitivity of 95.2% (95% CI: 86.9-98.4) and specificity of 57.1% (95% CI: 25.0-84.2). The yield was lower for lesions within 17 mm of the cortex: sensitivity of 74.6% (95% CI: 62.1-84.7) and specificity of 71.4% (95% CI: 29.0-96.3). Seven (10%) patients developed complications after the first biopsy and none after the second. CONCLUSIONS: The diagnostic yield was lower for lesions less than 2 cm in size and located superficially. In this series we did not observe an increased rate of complications after a second biopsy

[Frameless stereotactic biopsy: diagnostic yield and complications]. / Biopsia cerebral estereotáctica sin marco: capacidad diagnóstica y complicaciones.

OBJECTIVE: The aim of this study was to evaluate the variables that could modify the diagnostic yield of frameless stereotactic biopsy, as well as its complications. MATERIALS AND METHOD: This was a retrospective study of frameless stereotactic biopsies carried out between July 2008 and December 2011 at Donostia University Hospital. The variables studied were size, distance to the cortex, contrast uptake and location. RESULTS: A total of 70 patients were included (75 biopsies); 39 males and 31 females with an age range between 39 and 83 years. The total diagnostic yield in our series was 97.1%. For lesions >19mm, the technique offered a sensitivity of 95.2% (95% CI: 86.9-98.4) and specificity of 57.1% (95% CI: 25.0-84.2). The yield was lower for lesions within 17mm of the cortex: sensitivity of 74.6% (95% CI: 62.1-84.7) and specificity of 71.4% (95% CI: 29.0-96.3). Seven (10%) patients developed complications after the first biopsy and none after the second. CONCLUSIONS: The diagnostic yield was lower for lesions less than 2cm in size and located superficially. In this series we did not observe an increased rate of complications after a second biopsy.

Primary Ewing's sarcoma of the skull: radical resection and immediate cranioplasty after chemotherapy. A technical note.

PURPOSE: The current standard treatment of Ewing's sarcoma is chemotherapy followed by surgery, making an immediate cranial reconstruction in a one-step surgical procedure possible. METHODS: We describe the technique used to repair a cranial defect after the resection of a primary Ewing's sarcoma of the skull in a one-step surgical procedure. RESULTS: Bone repair with a custom-made cranioplasty immediately after resection of a primary Ewing's sarcoma of the skull avoids deformities and late complications associated with reconstructive surgery after radiotherapy and not interfere with radiotherapy and neither with follow-up. CONCLUSION: A one-step surgical procedure after chemotherapy for primary Ewing's sarcoma of the skull could be safer, less aggressive and more radical; avoiding deformities and late complications.

Elevation of oil body integrity and emulsion stability by polyoleosins, multiple oleosin units joined in tandem head-to-tail fusions.

We have successfully created polyoleosins by joining multiple oleosin units in tandem head-to-tail fusions. Constructs encoding recombinant proteins of 1, 3 and 6 oleosin repeats were purposely expressed both in planta and in Escherichia coli. Recombinant polyoleosins accumulated in the seed oil bodies of transgenic plants and in the inclusion bodies of E. coli. Although polyoleosin was estimated to only accumulate to <2% of the total oil body protein in planta, their presence increased the freezing tolerance of imbibed seeds as well as emulsion stability and structural integrity of purified oil bodies; these increases were greater with increasing oleosin repeat number. Interestingly, the hexameric form of polyoleosin also led to an observable delay in germination which could be overcome with the addition of external sucrose. Prokaryotically produced polyoleosin was purified and used to generate artificial oil bodies and the increase in structural integrity of artificial oil bodies-containing polyoleosin was found to mimic those produced in planta. We describe here the construction of polyoleosins, their purification from E. coli, and properties imparted on seeds as well as native and artificial oil bodies. A putative mechanism to account for these properties is also proposed.

The search for a role of the caudal intralaminar nuclei in the pathophysiology of Parkinson's disease.

The situation of the caudal intralaminar thalamic nuclei within basal ganglia circuits has gained increased attention over the past few years. Although initially considered as a "non-specific" thalamic nuclei, tract-tracing studies carried out over the past two decades have demonstrated that the centromedian-parafascicular thalamic complex (CM-Pf) is connected to virtually all basal ganglia components and related nuclei. Although the anatomical basis sustaining the thalamic modulation of basal ganglia circuits has long been characterized, the functional significance of these transverse circuits still remain to be properly accommodated within the basal ganglia model, both under normal conditions as well as in situations of dopaminergic depletion. However, the recent demonstration of primary (e.g., non-dopamine related) neurodegenerative phenomena restricted to the CM-Pf in Parkinson's disease (PD) has renewed interest in the role played by the caudal intralaminar nuclei in the pathophysiology of PD. Concomitantly, evidence has become available of increased metabolic activity in the caudal intralaminar nuclei in rodent models of PD. Finally, CM-Pf neurosurgery in patients suffering from PD has produced contrasting outcomes, indicating that a consensus is still to be reached regarding the potential usefulness of targeting the caudal intralaminar nuclei to treat movement disorders of basal ganglia origin.

High-resolution neuroanatomical tract-tracing for the analysis of striatal microcircuits.

Although currently available retrograde tracers are useful tools for identifying striatal projection neurons, transported tracers often remained restricted within the neuronal somata and the thickest, main dendrites. Indeed, thin dendrites located far away from the cell soma as well as post-synaptic elements such as dendritic spines cannot be labeled unless performing intracellular injections. In this regard, the subsequent use of anterograde tracers for the labeling of striatal afferents often failed to unequivocally elucidate whether a given afferent makes true contacts with striatal projections neurons. Here we show that such a technical constraint can now be circumvented by retrograde tracing using rabies virus (RV). Immunofluorescence detection with a monoclonal antibody directed against the viral phosphoprotein resulted in a consistent Golgi-like labeling of striatal projection neurons, allowing clear visualization of small-size elements such as thin dendrites as well as dendritic spines. The combination of this retrograde tracing together with dual anterograde tracing of cortical and thalamic afferents has proven to be a useful tool for ascertaining striatal microcircuits. Indeed, by taking advantage of the trans-synaptic spread of RV, different subpopulations of local-circuit neurons modulating striatal efferent neurons can also be identified. At the striatal level, structures displaying labeling were visualized under the confocal laser-scanning microscope at high resolution. Once acquired, confocal stacks of images were firstly deconvoluted and then processed through 3D-volume rendering in order to unequivocally identify true contacts between pre-synaptic elements (axon terminals from cortical or thalamic sources) and post-synaptic elements (projection neurons and/or interneurons labeled with RV).

Expression of vesicular glutamate transporters 1 and 2 in the cells of origin of the rat thalamostriatal pathway.

The present study is focused on the analysis of the vesicular glutamate transporters 1 and 2 (VGLUT1 and VGLUT2) used by thalamic neurons giving rise to the thalamostriatal system. Instead of studying the distribution of VGLUT proteins at the level of thalamostriatal terminals, this report is focused on identifying the expression of the VGLUT mRNAs within the parent cell bodies of thalamic neurons innervating the striatum. For this purpose, we have combined dual in situ hybridization to detect both VGLUT1 and VGLUT2 mRNAs together with retrograde tracing with cholera toxin. Our results show that VGLUT2 is the only vesicular glutamate transporter expressed in thalamostriatal-projecting neurons located in the midline and intralaminar nuclei, whereas all neurons from the ventral thalamic nuclei innervating the striatum express both VGLUTs, at least at the mRNA level. Indeed, the mRNAs encoding for VGLUT1 and VGLUT2 displayed a sharp complementary subcellular distribution within neurons from the ventral thalamic nuclei giving rise to thalamostriatal projections. The differential distribution of VGLUT mRNAs lead us to conclude that the thalamostriatal pathway is a dual system, composed by a preponderant projection arising from the midline and intralaminar nuclei using VGLUT2 as the glutamate transporter, together with another important source of striatal afferents arising from neurons in the ventral thalamic relay nuclei containing both kinds of vesicular glutamate transporters.

Detection of two different mRNAs in a single section by dual in situ hybridization: a comparison between colorimetric and fluorescent detection.

We have compared the performance of two methods designed to simultaneously detect two different mRNAs within a single brain section by dual ISH. Specific mRNA riboprobes labeled with biotin and digoxigenin were simultaneously hybridized and visualized using either brightfield or fluorescence microscopy. For brightfield visualization, the biotin-labeled riboprobe was detected with a peroxidase chromogen, whereas, an alkaline phosphatase substrate was used for the detection of the digoxigenin-labeled riboprobe. Dual fluorescent ISH involved the detection of the biotin-labeled riboprobe with an Alexa((R))488-conjugated streptavidin followed by the visualization of the digoxigenin-labeled riboprobe with the red fluorescent substrate HNPP. The dual ISH protocols presented here offer sensitive methods to detect the expression of two mRNAs of interest, with both colorimetric and fluorescent ISH each having its strengths and limitations. For example, dual colorimetric ISH has proven to be particularly useful to study the distribution of two mRNAs in different brain nuclei, whereas, dual fluorescent ISH has provided better results when studying the co-localization of two different mRNAs in single neurons. The comprehensive step-by-step procedure is presented, together with a troubleshooting section in which the advantages and limitations of these procedures are reviewed in depth. Moreover, alternative protocols for dual ISH were also compared to those presented here.

Expression of the mRNAs encoding for the vesicular glutamate transporters 1 and 2 in the rat thalamus.

Vesicular glutamate transporters (VGLUTs) are responsible for glutamate trafficking and for the subsequent regulated release of this excitatory neurotransmitter at the synapse. Three isoforms of the VGLUT have been identified, now known as VGLUT1, VGLUT2, and VGLUT3. Both VGLUT1 and VGLUT2 have been considered definitive markers of glutamatergic neurons, whereas VGLUT3 is expressed in nonglutamatergic neurons such as cholinergic striatal interneurons. It is widely believed that VGLUT1 and VGLUT2 are expressed in a complementary manner at the cortical and thalamic levels, suggesting that these glutamatergic neurons fulfill different physiological functions. In the present work, we analyzed the pattern of VGLUT1 and VGLUT2 mRNA expression at the thalamic level by using single and dual in situ hybridization. In accordance with current beliefs, we found significant expression of VGLUT2 mRNA in all the thalamic nuclei, while moderate expression of VGLUT1 mRNA was consistently found in both the principal relay and the association thalamic nuclei. Interestingly, individual neurons within these nuclei coexpressed both VGLUT1 and VGLUT2 mRNAs, suggesting that these individual thalamic neurons may have different ways of trafficking glutamate. These results call for a reappraisal of the previously held concept regarding the mutually exclusive distribution of VGLUT transporters in the central nervous system.

Thalamic innervation of the direct and indirect basal ganglia pathways in the rat: Ipsi- and contralateral projections.

The present study describes the thalamic innervation coming from the rat parafascicular nucleus (PF) onto striatal and subthalamic efferent neurons projecting either to the globus pallidus (GP) or to the substantia nigra pars reticulata (SNr) by using a protocol for multiple neuroanatomical tracing. Both striatofugal neurons targeting the ipsilateral SNr (direct pathway) as well as striatal efferent neurons projecting to the ipsilateral GP (indirect pathway) were located within the terminal fields of the thalamostriatal afferents. In the subthalamic nucleus (STN), both neurons projecting to ipsilateral GP as well as neurons projecting to ipsilateral SNr also appear to receive thalamic afferents. Although the projections linking the caudal intralaminar nuclei with the ipsilateral striatum and STN are far more prominent, we also noticed that thalamic axons could gain access to the contralateral STN. Furthermore, a small number of STN neurons were seen to project to both the contralateral GP and PF nuclei. These ipsi- and contralateral projections enable the caudal intralaminar nuclei to modulate the activity of both the direct and the indirect pathway.

Thalamic innervation of striatal and subthalamic neurons projecting to the rat entopeduncular nucleus.

The present study analyses the anatomical arrangement of the projections linking the Wistar rat parafascicular thalamic nucleus (PF) and basal ganglia structures, such as the striatum and the subthalamic nucleus (STN), by using neuroanatomical tract-tracing techniques. Both the thalamostriatal and the striato-entopeduncular projections were topographically organized, and several areas of overlap between identified circuits were noticed, sustaining the existence of up to three separated channels within the Nauta-Mehler loop. Thalamic afferents arising from dorsolateral PF territories are in register with striatofugal neurons located in dorsolateral striatal areas, which in turn project to dorsolateral regions of the entopeduncular nucleus (ENT). Medial ENT regions are innervated by striatal neurons located within medial striatal territories, these neurons being the target for thalamic afferents coming from medial PF areas. Finally, afferents from neurons located in ventrolateral PF areas approached striatal neurons in ventral and lateral striatal territories, which in turn project towards ventral and lateral ENT regions. Efferent STN neurons projecting to ENT were found to be the apparent postsynaptic target for thalamo-subthalamic axons. The thalamo-subthalamic projection was also topographically organized. Medial, central and lateral STN territories are innervated by thalamic neurons located within medial, ventrolateral and dorsolateral PF areas, respectively. Thus, each individual PF subregion projects in a segregated fashion to specific parts of the striato-entopeduncular and subthalamo-entopeduncular systems. These circuits enabled the caudal intralaminar nuclei to modulate basal ganglia output.