The use of unfixed bone marrow trephines for multicolour flow cytometry.

An adequate bone marrow aspirate is essential for a rapid diagnosis of acute leukaemia by multicolour flow cytometry enabling the simultaneous assessment of multiple antigens on the cell surface as well as intracellular or nuclear ones. In the context of acute leukaemia, it is important to have a diagnosis of the blasts lineage as soon as possible to decide the appropriate treatment. This is sometimes delayed due to difficulties in obtaining a bone marrow aspirate due to a "dry tap". In this study we evaluated retrospectively cell markers results by flow cytometry of unfixed bone marrow trephines of 65 patients with leukaemia at diagnosis and including a few after treatment. Our aims were: 1) To compare cell markers results between bone marrow trephine (BMT) and bone marrow aspirate (BMA) 24 cases and BMT with peripheral blood (PB) 14 cases in paired samples to establish if they were reproducible with results of the unfixed bone marrow trephine biopsies. 2) To ascertain a precise diagnosis in 27 (42%) of the cases in which only a bone marrow trephine was available. We demonstrated that unfixed bone marrow trephine provides an adequate and representative cell suspension for flow cytometry and it is a powerful tool when no other material (bone marrow aspirate or peripheral blood) is available to make a rapid diagnosis. Furthermore when marrow aspirate or peripheral blood paired samples were available, flow cytometry results obtained were identical across all the sample types. Applicability to the clinical laboratory: We described a method to obtain a cell suspension from core biopsies that can easily be implemented routinely in a laboratory that performs diagnostic flow cytometry immunophenotyping. This method is simple, inexpensive and it doesn't require extra equipment.

Outcome of surgery in children with focal cortical dysplasia younger than 5 years explored by stereo-electroencephalography.

PURPOSE: Focal cortical dysplasia (FCD) is the most frequent etiology for drug-resistant epilepsy in young children. Complete removal of the lesion is mandatory to cure the epilepsy. Stereo-EEG (SEEG) is an excellent method to delimitate the zone to be resected in older children and adults. We studied its feasibility in younger children. METHODS: We retrospectively studied 19 children under 5 years of age who underwent SEEG between January 2009 and December 2012 and were subsequently operated on. FCD was diagnosed in all. We reviewed magnetic resonance imaging (MRI), electrophysiological and clinical data, as well as postoperative seizure outcome. We also included fluoro-deoxyglucose positron emission tomography (FDG-PET) studies, which had been systematically performed before invasive recording in 16 of the 19 children. RESULTS: The mean patient's age at the time of SEEG was 38.6 months, and the mean age at seizure onset was 8 months. Three patients had normal MRI. No SEEG-associated complications occurred. We were able to delineate the epileptogenic zone in all children, and electrode stimulation localized the motor area when necessary (12 patients). Hypometabolic areas on FDG-PET included the epileptogenic zone in 13 of the 16 children, with a lobar concordance in 9 (56 %) and the same anatomical extent in 6 (38 %). Twelve children subsequently underwent focal or sublobar resection, six had multilobar resection, and one had hemispherotomy. The etiology was FCD type 2 in 15 and FCD type 1 or type 3 in three children. Eighty-four percent of our population have remained seizure-free at a mean follow-up of 29 months (12-48 months). CONCLUSION: Although children with FCD can successfully undergo resective surgery without invasive EEG, poor seizure semiology at this age inclines to perform SEEG when the dysplastic lesion is ill-defined and/or the electroclinical correlation is unclear. In cases with normal imaging as well as with suspected huge malformations, as was the case in 52 % of our patients, we consider it to be indispensable.

Upgrades to the gamma ray imager on DIII-D enabling access to high flux hard x-ray measurements during the runaway electron plateau phase (invited).

The Gamma Ray Imager (GRI) is a pinhole camera providing 2D imaging of MeV hard x-ray (HXR) bremsstrahlung emission from runaway electrons (REs) over the poloidal cross section of the DIII-D tokamak. We report a series of upgrades to the GRI expanding the access to RE scenarios from the diagnosis of a trace amount of REs to high flux HXR measurements during the RE plateau phase. We present the implementation of novel gamma ray detectors based on LYSO and YAP crystals coupled to multi-pixel photon counters, enabling a count rate in excess of 1 MHz. Finally, we highlight new insights into the RE physics discovered during the current quench and RE plateau phase experiments as the result of these upgrades.

Langmuir probe array for the small angle slot divertor in DIII-D.

The DIII-D small angle slot (SAS) divertor is designed for divertor physics studies with enhanced neutral confinement and special target geometries in a closed divertor. The closed nature of the SAS makes optical diagnostic measurements difficult, so a specially designed, multipurpose array of Langmuir probes has been implemented to study the plasma conditions in and around the slot. The probes are spaced to provide at least 2 mm resolution (shorter than the energy decay length) of the near scrape-off layer when mapped to the outer mid-plane. Due to space limitations at the bottom of the slot, a novel spring-loaded probe and tile design was developed to clamp several short rooftop probe tips and insulators to the cooled baseplate. Initial probe measurements revealed tile to tile edge shadowing, especially where magnetic field line surface angles were less than 1°. Additionally, it was found, using three Langmuir probes (at 90°, 180°, and 270°), that the strike point variation of ±5 mm radially around the torus was not well aligned with the circular slot geometry [Watkins et al., Nucl. Mater. Energy 18, 46 (2019)]. These issues were resolved by (1) designing tiles with all probes mounted near the tile center instead of near the edges and (2) aligning these new custom tiles to the measured strike point toroidal surface with a very accurate laser scanning alignment tool. Post-alignment Langmuir probe measurements and plasma behavior demonstrated close agreement at two separate toroidal locations that were 45° apart.

Adipocytes disrupt the translational programme of acute lymphoblastic leukaemia to favour tumour survival and persistence.

The specific niche adaptations that facilitate primary disease and Acute Lymphoblastic Leukaemia (ALL) survival after induction chemotherapy remain unclear. Here, we show that Bone Marrow (BM) adipocytes dynamically evolve during ALL pathogenesis and therapy, transitioning from cellular depletion in the primary leukaemia niche to a fully reconstituted state upon remission induction. Functionally, adipocyte niches elicit a fate switch in ALL cells towards slow-proliferation and cellular quiescence, highlighting the critical contribution of the adipocyte dynamic to disease establishment and chemotherapy resistance. Mechanistically, adipocyte niche interaction targets posttranscriptional networks and suppresses protein biosynthesis in ALL cells. Treatment with general control nonderepressible 2 inhibitor (GCN2ib) alleviates adipocyte-mediated translational repression and rescues ALL cell quiescence thereby significantly reducing the cytoprotective effect of adipocytes against chemotherapy and other extrinsic stressors. These data establish how adipocyte driven restrictions of the ALL proteome benefit ALL tumours, preventing their elimination, and suggest ways to manipulate adipocyte-mediated ALL resistance.

Optimizing the Detection of Subtle Insular Lesions on MRI When Insular Epilepsy Is Suspected.

Insular epilepsy is underdiagnosed and accounts for a number of failed operations. Identifying insular target lesions on MR imaging can help guide intracranial electroencephalography and improve the outcome of surgery. In this study, we present a novel method of exploring the insular region for subtle lesions on 3D MR imaging by MPR postprocessing of slices in oblique reference planes. Using this method, we retrospectively reviewed presurgical MRIs that were initially considered to have normal findings in 7 pediatric patients with intractable insular epilepsy. Insular epilepsy was confirmed in these patients on stereo-electroencephalography and histopathology. The MPR postprocessing method we describe helped detect subtle insular lesions in all 7 patients.

Thomson scattering measurements on DIII-D using in-vessel laser mirrors and lenses to diagnose a new divertor location.

Translatable in-vessel mirrors have enabled the DIII-D Thomson scattering system to diagnose the divertor plasma in high triangularity shaped plasmas. Previous divertor Thomson scattering measurements in DIII-D were restricted to spatial locations along a Nd:YAG laser beam that was directed through a vertical port. This only allowed measurements to be made in low triangularity shaped plasmas. The new mirrors re-route the laser underneath floor tiles to a position of smaller major radius as necessary for high triangularity plasmas. New in-vessel collection optics transmit scattered light from regions inaccessible to external lenses. Damage to mirrors and high stray light levels are challenges that were overcome to successfully make these measurements. Through the careful use of baffles and light shields, stray light leakage into polychromator detector channels was reduced to negligible levels, allowing temperature measurements below 1 eV. The system is described and the initial results presented.

Faraday-effect polarimeter diagnostic for internal magnetic field fluctuation measurements in DIII-D.

Motivated by the need to measure fast equilibrium temporal dynamics, non-axisymmetric structures, and core magnetic fluctuations (coherent and broadband), a three-chord Faraday-effect polarimeter-interferometer system with fast time response and high phase resolution has recently been installed on the DIII-D tokamak. A novel detection scheme utilizing two probe beams and two detectors for each chord results in reduced phase noise and increased time response [δb ∼ 1G with up to 3 MHz bandwidth]. First measurement results were obtained during the recent DIII-D experimental campaign. Simultaneous Faraday and density measurements have been successfully demonstrated and high-frequency, up to 100 kHz, Faraday-effect perturbations have been observed. Preliminary comparisons with EFIT are used to validate diagnostic performance. Principle of the diagnostic and first experimental results is presented.

Upgraded divertor Thomson scattering system on DIII-D.

A design to extend the unique divertor Thomson scattering system on DIII-D to allow measurements of electron temperature and density in high triangularity plasmas is presented. Access to this region is selectable on a shot-by-shot basis by redirecting the laser beam of the existing divertor Thomson system inboard - beneath the lower floor using a moveable, high-damage threshold, in-vacuum mirror - and then redirecting again vertically. The currently measured divertor region remains available with this mirror retracted. Scattered light is collected from viewchords near the divertor floor using in-vacuum, high temperature optical elements and relayed through the port window, before being coupled into optical fiber bundles. At higher elevations from the floor, measurements are made by dynamically re-focusing the existing divertor system collection optics. Nd:YAG laser timing, analysis of the scattered light spectrum via polychromators, data acquisition, and calibration are all handled by existing systems or methods of the current multi-pulse Thomson scattering system. Existing filtered polychromators with 7 spectral channels are employed to provide maximum measurement breadth (Te in the range of 0.5 eV-2 keV, ne in the range of 5 × 1018-1 × 1021 m3) for both low Te in detachment and high Te measurement up beyond the separatrix.

Gamma ray imager on the DIII-D tokamak.

A gamma ray camera is built for the DIII-D tokamak [J. Luxon, Nucl. Fusion 42, 614 (2002)] that provides spatial localization and energy resolution of gamma flux by combining a lead pinhole camera with custom-built detectors and optimized viewing geometry. This diagnostic system is installed on the outer midplane of the tokamak such that its 123 collimated sightlines extend across the tokamak radius while also covering most of the vertical extent of the plasma volume. A set of 30 bismuth germanate detectors can be secured in any of the available sightlines, allowing for customizable coverage in experiments with runaway electrons in the energy range of 1-60 MeV. Commissioning of the gamma ray imager includes the quantification of electromagnetic noise sources in the tokamak machine hall and a measurement of the energy spectrum of background gamma radiation. First measurements of gamma rays coming from the plasma provide a suitable testbed for implementing pulse height analysis that provides the energy of detected gamma photons.

Durable remissions of myelodysplastic syndrome and acute myeloid leukemia after reduced-intensity allografting.

PURPOSE: To evaluate the use of reduced-intensity (RI) conditioning with allogeneic hematopoietic stem cell transplantation (HSCT) from HLA-identical family donors in patients with myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML). PATIENTS AND METHODS: Sixteen patients (median age, 54 years; range, 37 to 66 years) underwent RI-HSCT using a conditioning regimen of fludarabine 25 mg/m2 daily for 5 days and either cyclophosphamide 1 g/m2 daily for 2 days (14 patients) or melphalan 140 mg/m2 for 1 day (two patients). The median number of CD34+ cells and CD3+ cells infused per kilogram of recipient weight was 4.5 x 106 (range, 1.8 to 7.3 x 106 cells) and 2.9 x 108 (range, 0.1 to 9.6 x 108 cells), respectively. RESULTS: There was no transplant-related mortality (TRM) within 100 days of HSCT. Grade 1 to 2 acute graft-versus-host disease (GVHD) occurred in three patients, but neither grade 3 nor grade 4 disease was observed. Chronic GVHD occurred in 10 patients. One patient had cytomegalovirus (CMV) reactivation but did not develop CMV disease. With a median follow-up of 26 months (range, 15 to 45 months), 11 patients are alive (nine in continuous complete remission and one in complete remission after a second transplantation), and five have died (four from disease progression and one from bone-marrow aplasia induced by cyclosporine withdrawal). The 2-year actuarial overall and event-free survival rates were 69% (95% confidence interval [CI], 40% to 86%) and 56% (95% CI, 30% to 68%), respectively. CONCLUSION: This strategy of RI-HSCT resulted in reliable engraftment with low incidence of acute GVHD and TRM. Durable remissions were observed in patients with MDS and AML consistent with a graft-versus-leukemia effect.

Invasive EEG explorations.

The Wada test was adapted from the procedure described by Wada in 1964. It still has a role in the prognostic evaluation of memory disorders after mesial temporal lobectomy. The test consists of injecting a short-acting anesthetic into one hemisphere, under continuous EEG monitoring and during carotid catheterization, to verify the function of contralateral structures. Intracranial EEG recordings deliver signals with few artifacts, and which are quite specific of the zone explored. Three types of electrodes are in common use: (a) foramen ovale (FO) electrodes: electrodes can be inserted directly, without any stereotactic procedure, to provide easy and comparative EEG recordings of the lower and middle portions of the temporal lobe close to the hippocampus. These allow validation of the temporal lobe origin of seizures using FO electrodes recording coupled with scalp EEG; (b): subdural strip or grip electrodes. This relatively aggressive technique carries infectious and hemorrhagic risks and does not allow the exploration of deep cortical structures. However, it permits precise functional cortical mapping via electrical stimulation because of dense and regular positioning of electrodes over the cortical convexity; (c) stereotactically implanted depth electrodes (stereo-electroencephalography [SEEG]). Electrodes are individually planned and inserted within the brain parenchyma through small burr holes. This technique is less aggressive than subdural grid exploration. However it offers relatively limited spatial sampling that may be less well adapted to precise functional evaluation. It allows recording from deep cortical structures and can be argued to be the gold standard of presurgical EEG exploration.

Long-term EEG in children.

Long-term video-EEG corresponds to a recording ranging from 1 to 24 h or even longer. It is indicated in the following situations: diagnosis of epileptic syndromes or unclassified epilepsy, pre-surgical evaluation for drug-resistant epilepsy, follow-up of epilepsy or in cases of paroxysmal symptoms whose etiology remains uncertain. There are some specificities related to paediatric care: a dedicated pediatric unit; continuous monitoring covering at least a full 24-hour period, especially in the context of pre-surgical evaluation; the requirement of presence by the parents, technician or nurse; and stronger attachment of electrodes (cup electrodes), the number of which is adapted to the age of the child. The chosen duration of the monitoring also depends on the frequency of seizures or paroxysmal events. The polygraphy must be adapted to the type and topography of movements. It is essential to have at least an electrocardiography (ECG) channel, respiratory sensor and electromyography (EMG) on both deltoids. There is no age limit for performing long-term video-EEG even in newborns and infants; nevertheless because of scalp fragility, strict surveillance of the baby's skin condition is required. In the specific context of pre-surgical evaluation, long-term video-EEG must record all types of seizures observed in the child. This monitoring is essential in order to develop hypotheses regarding the seizure onset zone, based on electroclinical correlations, which should be adapted to the child's age and the psychomotor development.

Evaluation of four candidate genes encoding proteins of the dopamine pathway in familial and sporadic Parkinson's disease: evidence for association of a DRD2 allele.

We assessed the role of four candidate genes encoding proteins involved in dopaminergic transmission, the dopamine transporter (DAT), the dopamine receptor D2 (DRD2), and the main catabolic enzymes of dopamine, monoamine oxidase A (MAOA) and B (MAOB), through allelic association studies in a population of familial and sporadic Parkinson's disease (PD). Using intronic polymorphisms of the four candidate genes, we studied the allelic distributions of the polymorphic markers in 18 affected members, one patient was chosen randomly from each PD family; 60 sporadic PD and 60 healthy unrelated control subjects were matched for sex and for country of origin. All subjects were white. To complete the study of the DRD2, we subsequently tested 40 additional sporadic PD and 40 control patients, who were recruited using a similar procedure. For DAT, MAOA, MAOB polymorphisms, similar allelic frequencies were present in familial, sporadic PD and control patients. In contrast, at the DRD2 locus, the overall allelic distribution was significantly different in the sporadic PD (p < 0.01) and in the familial PD groups (p < 0.05), each was compared with the controls. The odd ratios were significant (p < 0.01) in sporadic PD and in familial PD for allele 3 with respective values of 1.84 (95% CI, 1.23-2.74) and 2.83 (95% CI, 1.32-6.08). Individuals who were homozygous for allele 3 were 2.3 times more frequent in the sporadic PD than in controls. Results suggest that DRD2, but not DAT, MAOA and MAOB, might be a genetic determinant of PD in the population tested.

Sternal osteomyelitis caused by Aspergillus fumigatus in a patient with previously treated Hodgkin's disease.

This report details the case of a 67 year old woman with sternal osteomyelitis caused by Aspergillus fumigatus. She was diagnosed with Hodgkin's disease in 1975 and was successfully treated with chemotherapy. A lobectomy for recurrence localised to the left lung was complicated nine years later by severe bronchiectasis, for which she required a total left sided pneumonectomy. At surgery, a non-invasive aspergillus was found. She presented eight years later with symptoms that were initially attributed to recurrence of Hodgkins's disease, but on investigation were found to be caused by fungal sternal osteomyelitis. Treatment with itraconazole suspension at a dose of 400 mg daily was successful.

A clinical and genetic study of familial cases of Parkinson's disease.

We assessed a group of patients with a family history of Parkinson's disease (PD) in order to see if they differed clinically from sporadic cases and to study their genetic characteristics. Index cases were selected on the basis of clinically typical PD, and at least one affected relative. Fourteen families including 110 first degree and 40 second degree relatives were ascertained. A total of 31 individuals (17 females and 14 males) were found to be affected. This group was compared for selected clinical parameters to 31 age matched patients with sporadic PD. No statistical difference was found between the two groups. In familial cases, both the clinical parameters studied and the course of the disease varied within and between families, as observed in sporadic cases. The genetic transmission was compatible with an autosomal dominant model. The total segregation ratio of 0.25 suggested an incomplete penetrance, which increased with age, from 0 below the age of 30 to 0.43 over the age of 70. Age at onset was earlier in children than in their parents in the 8 multigeneration kindreds studied (mean difference 26 +/- 4.6 years, p = 0.01), whereas it was identical within a generation (mean difference 4.7 +/- 5.7 years, p = 0.1). Although we cannot exclude an ascertainment bias, our results are compatible with an anticipation phenomenon, which deserves further studies for confirmation.

Gelastic seizures: video-EEG and scintigraphic analysis of a case with a frontal focus; review of the literature and pathophysiological hypotheses.

We report scalp EEG and SPECT findings in a young patient who experienced gelastic seizures; clinical, EEG and scintigraphic data strongly suggested a frontal focus in a context of cryptogenic epilepsy. Few cases of gelastic seizures originating in the frontal lobe have been reported in the literature, most of them involving a diencephalic hamartoma or a temporal focus although, no clinical pattern has been found to be specific for each of these three anatomical regions. The ictal laughter is of variable nature, unmotivated or associated with feelings of mirth, forced or natural, except in the case of a frontal focus where the laughter seems consistently described as forced and unmotivated. However, mirth and laughter are two dissociable clinical elements; their genesis probably involves distinct mechanisms. Anatomical considerations lead to several hypotheses concerning laughter generation: it could be a simple reactional behavior in response to a modified cognitive process, an automatic behavior or a forced action. In a few cases with a temporal focus, laughter seems directly related to a disorganization of the associative temporal cortex and may be considered as a reactional behavior. In cases with frontal focus, anterior cingulate and orbital structures would be particularly implicated in laughter genesis, although with possible different pathophysiological routes: in the first case by disconnection within the premotor mesial system or by an imbalance between premotor mesial and premotor lateral systems, and in the second case by activation of a previously conditioned orbital region.

Video-EEG and ictal SPECT in three patients with both epileptic and non-epileptic seizures.

We report scalp EEG and ictal SPECT findings in epileptic (complex partial) and non-epileptic seizures in three patients who experienced both types of event during presurgical investigation of medically intractable epilepsies. In all three patients, ictal SPECT showed localizing changes in cerebral blood flow during epileptic seizures, but showed no change during pseudoseizures. In two patients, the physical manifestations of the pseudoseizures were similar to those of the epileptic seizures, supporting the contention that physiological activation is unlikely to mimic ictal perfusion changes. In one patient, the EEG recording was rendered difficult to interpret by muscle artefact, while SPECT was clear and showed no change. SPECT is not a primary tool for diagnosis of pseudoseizures, but when patients undergoing presurgical investigation are injected during pseudoseizures, then SPECT is unlikely to show misleading perfusion changes due to activation effects, and may aid diagnosis where there is muscle artefact on EEG.

[Indications and risks of neurosurgical techniques for drug-resistant partial epilepsy in adults]. / Indications et risques des techniques neurochirurgicales chez l'adulte pour l'épilepsie partielle pharmaco-résistante.

Many different surgical procedures are performed for medically refractory partial epilepsy. Some surgical therapies are performed to cure the epilepsy (for example unifocal epilepsy), others are palliative procedures. To cure epilepsy, temporal lobectomy is the most common surgical procedure. The different techniques are shortly described, indications and complications are discussed.

[Atypical familial parkinsonian syndromes. Parkinson diseases or specific entities?]. / Les syndromes parkinsoniens familiaux "atypiques". Maladies de Parkinson ou entités autonomes?

RECENT DESCRIPTION: Recent report have described "atypical" familial extrapyramidal syndromes similar to authentic Parkinson's disease and well-defined genetic diseases. PERRY SYNDROME: Onset occurs between 35 and 57 years, leading to death within 3 to 7 years. The syndrome associates a Parkinson's syndrome, athymormia and hypoventilation. Massive neuronal depopulation in the locus niger and rare Lewy bodies are seen. PARKINSON'S SYNDROME WITH PERIPHERAL NEUROPATHY: In addition to the extrapyramidal signs, there is ptosis, neuropathy and sometimes dementia and major neurone loss in the locus niger. No Lewy bodies have been identified. PARKINSON'S SYNDROME WITH PALLIDOPONTONIGRAL DEGENERATION: Onset occurs between 32 and 58 years, leading to death within 8 years. Extrapyramidal signs, falls, supranuclear palsy and dementia are observed. Neurone loss is severe in the pars compacta, locu sniger, palladium, pons, and mesencephalic tegmentum. There are no Lewy bodies. EARLY-ONSET PARKINSON'S SYNDROME: Beginning between 2 and 39 years, there are no associated neurological signs. Severe neurone loss in the pars compacta and the pars reticulata of the niger locus without Lewy bodies. PARKINSON'S SYNDROME-DEMENTIA WITH "BALLOON NEURONES": This syndrome begins at 24-59 years and leads to death in 8 to 11 years. There are extrapyramidal signs, a pyramidal syndrome, dementia, generalized seizures and dysautonomia. Major neurone loss occurs with balloon neurones in the anterior temporal cortex, the amygdala, the parahippocampal gyrus, the hypothalamus, the dorsal nucleus of the X and rare Lewy bodies. PARKINSON'S SYNDROME FRONTAL DEMENTIA AND AMYOTROPHY: Beginning between 27 and 56 years, the syndrome leads to death in 13 years and associates frontal dementia with motor neurone defects with the extrapyramidal signs. There is neurone loss in the locus niger and amygdala as well as in the anterior horn of the cord. There are no Lewy bodies. SPECIFIC CLINICOPATHOLOGICAL ENTITIES: is the most likely hypothesis. There is no anatomoclinical evidence suggesting these syndromes should be considered to be Parkinson's disease.