cometadata/datacite-titles-descriptions-related-identifiers huggingface dataset:
DOI: 10.6084/m9.figshare.5104690
title: Supplementary Material for: Effect of Arsenic Trioxide on Uveal Melanoma Cell Proliferation in vitro

description: <i>Aims:</i> The aim of this study was to investigate the effect of arsenic trioxide (ATO) on the growth of cultured uveal melanoma cells (OCM-1 cell line). <i>Methods:</i> Cultured OCM-1 cells were treated with 0.75–24 µ<i>M</i> of ATO for 4–96 h. Cell viability was evaluated with the methylthiazoletetrazolium (MTT) assay as proliferation test. Apoptotic and necrotic cells were quantified using flow cytometry following Annexin-V/PI double stain. The cell morphology alteration was examined by light and electron transmission microscopy. To investigate the underlying mechanism of ATO-induced apoptosis and necrosis, glutathione peroxidase activity was measured, and mitochondrial membrane potentials were quantified using confocal microscopy. <i>Results:</i> In the MTT assays, OCM-1 cell growth was inhibited at ATO concentrations of 1.5–24 µ<i>M</i>. The 50% inhibitory concentrationof ATO was 16.8 µ<i>M</i>. At ATO concentrations of 12 and 24 µ<i>M</i>, apoptosis and necrosis were induced after 24 h of incubation as shown by light and transmission electron microscopy. At ATO concentrations of 12 and 24 µ<i>M</i>, the glutathione peroxidase activity was significantly (p &lt; 0.05) reduced after 24 h of incubation, and the mitochondrial membrane potentials were significantly (p &lt; 0.01) decreased after 4, 12 and 24 h of incubation. <i>Conclusion:</i> ATO inhibited ocular melanoma cell line growth at concentrations of 1.5–24 µ<i>M</i> of ATO in a dose- and time-dependent manner by inducing apoptosis and necrosis of the tumor cells. The concurringly decreased glutathione peroxidase activity and the reduced mitochondrial membrane potentials may be possible underlying mechanisms for the apoptosis and necrosis of the tumor cells.
DOI: 10.6084/m9.figshare.5104693.v1
title: Supplementary Material for: Clinical MRI Cutoff Points for Predicting Lacunar Stroke May Not Exist: Need for a Grading rather than a Dichotomizing System

description: <i>Background:</i> Lacunar stroke has been defined as an infarct &lt;15 mm in diameter in the presence of symptoms of lacunar syndromes. We investigated a new approach in predicting whether a deep infarct is caused by small arterial occlusion. <i>Methods:</i> A total of 319 consecutive patients with acute symptomatic infarcts within the striatocapsular territory underwent diffusion-weighted imaging (DWI) and diagnostic workups, including vascular and cardiological studies. Predictors for nonlacunar mechanisms were evaluated by logistic regression analysis, with the size of infarct (1-mm increase) and stroke syndrome (traditional vs. atypical lacunar syndrome vs. cortical syndrome) graded rather than dichotomized. <i>Results:</i> Amongst the 171 patients who did not meet the established criteria for lacunar stroke, that is, deep infarct of ≧15 mm or presenting symptoms of nontraditional lacunar syndrome, a documented etiology could not be determined in 97 (56.7%) patients. In contrast, amongst the 148 patients who met the criteria, 27 (18.2%) had nonlacunar mechanisms. Logistic regression analysis identified the variables that predicted nonlacunar stroke mechanisms with statistical significance as nontraditional lacunar syndromes (OR 2.19 for atypical lacunar syndrome, and OR 6.72 for cortical syndrome), infarct size on DWI (OR 1.05 per 1-mm increase), and unilateral multiple deep infarcts on DWI (OR 2.22, p &lt; 0.05 in all cases). Receiver operating characteristic curves showed that discrimination power of the model derived from logistic regression analysis (grading system) was better than that of the previously established dichotomizing criterion in predicting nonlacunar mechanisms (p = 0.004). <i>Conclusions: </i>A clinically significant proportion of clinical MRI lacunae are associated with underlying nonlacunar mechanisms. Decisions regarding the extent of diagnostic procedures in patients with subcortical infarcts can be guided by the point value in terms of the stroke syndrome and infarct patterns, as well as the size of infarct.
DOI: 10.6084/m9.figshare.5104684
title: Supplementary Material for: Microtubule-Associated Protein 1B Interaction with Tubulin Tyrosine Ligase Contributes to the Control of Microtubule Tyrosination

description: Microtubule-associated protein 1B (MAP1B) is the first microtubule-associated protein to be expressed during nervous system development. MAP1B belongs to a large family of proteins that contribute to the stabilization and/or enhancement of microtubule polymerization. These functions are related to the control of the dynamic properties of microtubules. The C-terminal domain of the neuronal α-tubulin isotype is characterized by the presence of an acidic polypeptide, with the last amino acid being tyrosine. This tyrosine residue may be enzymatically removed from the protein by an unknown carboxypeptidase activity. Subsequently, the tyrosine residue is again incorporated into this tubulinby another enzyme, tubulin tyrosine ligase, to yield tyrosinated tubulin. Because neurons lacking MAP1B have a reduced proportion of tyrosinated microtubules, we analyzed the possible interaction between MAP1B and tubulin tyrosine ligase. Our results show that these proteins indeed interact and that the interaction is not affected by MAP1B phosphorylation. Additionally, neurons lacking MAP1B, when exposed to drugs that reversibly depolymerize microtubules, do not fully recover tyrosinated microtubules upon drug removal. These results suggest that MAP1B regulates tyrosination of α-tubulin in neuronal microtubules. This regulation may be important for general processes involved in nervous system development such as axonal guidance and neuronal migration.
DOI: 10.6084/m9.figshare.5104702.v1
title: Supplementary Material for: Gene Expression Profiling of Hypothalamic Hamartomas: A Search for Genes Associated with Central Precocious Puberty

description: <i>Background:</i> Hypothalamic hamartomas (HHs) are congenital lesions composed of neurons and astroglia. Frequently, HHs cause central precocious puberty (CPP) and/or gelastic seizures. Because HHs might express genes similar to those required for the initiation of normal puberty, we used cDNA arrays to compare the gene expression profile of an HH associated with CPP with three HHs not accompanied by sexual precocity. <i>Methods:</i> Global changes in gene expression were detected using Affymetrix arrays. The results were confirmed by semiquantitative PCR, which also served to examine the expression of selected genes in the hypothalamus of female monkeys undergoing puberty. <i>Results:</i> All HHs were associated with seizures. Ten genes whose expression was increased in the HH with CPP were identified. They encode proteins involved in three key cellular processes: transcriptional regulation, cell-cell signaling, and cell adhesiveness. They include <i>IA-1</i> and <i>MEF2A</i>, two transcription factors required for neuronal development; <i>mGluR1</i> and<i> VILIP-1</i>, which encode proteins involved in neuronal communication, and <i>TSG-6 </i>that encodes a protein involved in cell adhesiveness. Of these, expression of <i>mGluR1</i> also increases in the female monkey hypothalamus at puberty. <i>Conclusions:</i> Increased expression of these genes in HHs may be relevant to the ability of some HHs to induce sexual precocity.
DOI: 10.6084/m9.figshare.5104678
title: Supplementary Material for: Nocodazole-Induced Changes in Microtubule Dynamics Impair the Morphology and Directionality of Migrating Medial Ganglionic Eminence Cells

description: We have shown previously that actomyosin contractility plays an important role in controlling nuclear movements in future interneurons born in the medial ganglionic eminence (MGE) [Bellion et al.: J Neurosci 2005;25:5691–5699]. Because microtubules are known to control the structural and motile properties of migrating neurons, we asked whether alterations in the dynamic instability of microtubules would impair MGE cell migration. Migration was analyzed in flat cocultures in which green-fluorescent-protein-expressing MGE cells migrate on cortical cells from their explant of origin. A low (100 n<i>M</i>) concentration of nocodazole shortened the leading process of MGE cells that nevertheless continued to migrate at the same rate but frequently changed their direction of migration relative to control cells. MGE cells treated with a higher (1 µ<i>M</i>) concentration of nocodazole that strongly destabilized microtubules took on multipolar morphology. They extended thin and labile processes. MGE cells no longer exhibited directional migration and migration velocity slowed 2-fold. These results suggest that microtubule stability is crucial for maintaining polarity and controlling the directional migration of MGE cells, whereas additional mechanisms are required to control cell motility.
DOI: 10.6084/m9.figshare.5104681
title: Supplementary Material for: Alternative Transcripts of <i>Dclk</i>1 and <i>Dclk</i>2 and Their Expression in <i>Doublecortin</i> Knockout Mice

description: The <i>doublecortin</i> (<i>DCX</i>) gene, mutated in X-linked human lissencephaly, has 2 close paralogs, <i>doublecortin-like kinase</i> 1 and 2 (<i>Dclk</i>1 and 2). In this study we attempted to better understand the dramatic differences between human and mouse DCX/Dcx-deficient phenotypes, focusing on the <i>Dclk</i> genes which are likely to compensate for Dcx function in the mouse. Using sequence database screens, Northern blot analyses and in situ hybridization experiments, we characterized the developmental transcripts of <i>Dclk</i>1 and 2, questioning their conservation between mouse and human, and their similarity to <i>Dcx</i>. Like <i>Dcx</i>, <i>Dcx</i>-like transcripts of the <i>Dclk</i>1 gene are expressed in postmitotic neurons in the developing cortex. No changes of expression were observed at the RNA level for these transcripts in <i>Dcx</i> knockout mice. However, a minor change in expression at the protein level was detected. The <i>Dclk</i>2 gene is less well characterized than <i>Dclk1 </i>andwe show here that it is expressed both in proliferating cells and postmitotic neurons, with a notably strong expression in the ventral telencephalon. No major differences in <i>Dclk</i>2 expression at the RNA and protein levels were identified comparing <i>Dcx</i> knockout and wild-type brains. We also analyzed <i>Dclk</i>1 and 2 expression in the hippocampal CA3 region which, unlike the neocortex, is abnormal in <i>Dcx</i> knockout mice. Interestingly, each transcript was expressed in CA3 neurons, including in the heterotopic pyramidal layer of <i>Dcx</i> knockout animals, but is presumably not able to compensate for a lack of <i>Dcx</i>. These results, in addition to characterizing the transcript diversity of an important family of genes, should facilitate further studies of compensation in <i>Dcx</i>-deficient mice.
DOI: 10.6084/m9.figshare.5104690.v1
title: Supplementary Material for: Effect of Arsenic Trioxide on Uveal Melanoma Cell Proliferation in vitro

description: <i>Aims:</i> The aim of this study was to investigate the effect of arsenic trioxide (ATO) on the growth of cultured uveal melanoma cells (OCM-1 cell line). <i>Methods:</i> Cultured OCM-1 cells were treated with 0.75–24 µ<i>M</i> of ATO for 4–96 h. Cell viability was evaluated with the methylthiazoletetrazolium (MTT) assay as proliferation test. Apoptotic and necrotic cells were quantified using flow cytometry following Annexin-V/PI double stain. The cell morphology alteration was examined by light and electron transmission microscopy. To investigate the underlying mechanism of ATO-induced apoptosis and necrosis, glutathione peroxidase activity was measured, and mitochondrial membrane potentials were quantified using confocal microscopy. <i>Results:</i> In the MTT assays, OCM-1 cell growth was inhibited at ATO concentrations of 1.5–24 µ<i>M</i>. The 50% inhibitory concentrationof ATO was 16.8 µ<i>M</i>. At ATO concentrations of 12 and 24 µ<i>M</i>, apoptosis and necrosis were induced after 24 h of incubation as shown by light and transmission electron microscopy. At ATO concentrations of 12 and 24 µ<i>M</i>, the glutathione peroxidase activity was significantly (p &lt; 0.05) reduced after 24 h of incubation, and the mitochondrial membrane potentials were significantly (p &lt; 0.01) decreased after 4, 12 and 24 h of incubation. <i>Conclusion:</i> ATO inhibited ocular melanoma cell line growth at concentrations of 1.5–24 µ<i>M</i> of ATO in a dose- and time-dependent manner by inducing apoptosis and necrosis of the tumor cells. The concurringly decreased glutathione peroxidase activity and the reduced mitochondrial membrane potentials may be possible underlying mechanisms for the apoptosis and necrosis of the tumor cells.
DOI: 10.6084/m9.figshare.5104675
title: Supplementary Material for: Stromal-Derived Factor 1 Signalling Regulates Radial and Tangential Migration in the Developing Cerebral Cortex

description: Stromal-derived factor 1 (SDF-1), a known chemoattractant, and its receptor CXCR4 are widely expressed in the developing and adult cerebral cortex. Recent studies have highlighted potential roles for SDF-1 during early cortical development. In view of the current findings, our histological analysis has revealed a distinct pattern of SDF-1 expression in the developing cerebral cortex at a time when cell proliferation and migration are at peak. To determine the role of chemokine signalling during early cortical development, embryonic rat brain slices were exposed to a medium containing secreted SDF-1 to perturb the endogenous levels of chemokine. Alternatively, brain slices were treated with 40 µ<i>M</i> of T140 or AMD3100, known antagonists of CXCR4. Using these experimental approaches, we demonstrate that chemokine signalling is imperative for the maintenance of the early cortical plate. In addition, we provide evidence that both neurogenesis and radial migration are concomitantly regulated by this signalling system. Conversely, interneurons, although not dependent on SDF-1 signalling to transgress the telencephalic boundary, require the chemokine to maintain their tangential migration. Collectively, our results demonstrate that SDF-1 with its distinct pattern of expression is essential and uniquely positioned to regulate key developmental events that underlie the formation of the cerebral cortex.
DOI: 10.6084/m9.figshare.5104672
title: Supplementary Material for: Migratory Behavior of Presumptive Interneurons Is Affected by AMPA Receptor Activation in Slice Cultures of Embryonic Mouse Neocortex

description: It has been reported that functional α-amino-3-hydroxy-5-methyl-isoxazolpro-prionic acid (AMPA) receptors permeable to calcium are already expressed by tangentially migrating prospective interneurons in the developing rodent cerebral cortex. However, the role of these receptors is not yet fully understood. To examine the effect of activation of AMPA receptors on tangential migration, we exposed migrating prospective interneurons derived from the medial ganglionic eminence (MGE) to AMPA in slice cultures and performed time lapse imaging. In the neocortex, a subpopulation of MGE-derived cells stopped migration or changed the direction of migration in response to AMPA in a dose-dependent manner. In contrast, neither MGE-derived cells migrating in the subcortical territory nor radially migrating cells in the neocortex were affected by exposure to AMPA. Transfection of dominant-negative AMPA receptor subunit GluR1 to the tangentially migrating cells prevented the effects of AMPA on migration. This study provides evidence that the activation of AMPA receptors can directly affect tangential migration in the neocortical part of telencephalic slice cultures.
DOI: 10.6084/m9.figshare.5104666
title: Supplementary Material for: How Neuronal Migration Contributes to the Morphogenesis of the CNS: Insights from the Zebrafish

description: We used transgenic zebrafish expressing GFP or YFP in subpopulations of neurons to study the migration, homing process and axon extension of groups of CNS neurons in different regions of the zebrafish brain. We found that extensive migration takes place at all levels of the CNS and gives rise to nuclei or cell populations with specific identities. Here, we describe 4 previously unknown or only partially characterized migratory events taking place in the zebrafish telencephalon and rhombic lip, using 3 different transgenic lines, and identify the phenotypes of the cells undertaking these migrations. The migration of a subgroup of mitral cell precursors from the dorsocaudal telencephalon to the olfactory bulb, visualized in the <i>tg(tbr1:YFP)</i> transgenic line, is coupled with morphogenetic transformation of the dorsal telencephalon. The <i>tg(1.4dlx5a-6a:GFP)</i> transgenic line provides a means to analyze the migration of GABAergic interneurons from the ventral to the dorsal telencephalon, thus extending the occurrence of this migration to another vertebrate. The <i>tg(Xeom:GFP)</i> transgenic line provides the first demonstration of the dorsoventral migration of glutamatergic septal neurons, present in mammals and now described in fish, thus reconciling the contrasting evidence of dorsal patterning genes <i>(tbr1, eomes)</i> expressed in a ventral cell population. Furthermore, migration studies in the <i>tg(1.4dlx5a-6a:GFP)</i> and <i>tg(Xeom:GFP)</i> lines help determine the origin of 2 important cell populations in the fish cerebellum: projection neurons and Purkinje cells. These examples reinforce the concept that migratory events contribute to the distribution of cell types with diverse identities through the CNS and that zebrafish transgenic lines represent excellent tools to study these events.
DOI: 10.6084/m9.figshare.5104672.v1
title: Supplementary Material for: Migratory Behavior of Presumptive Interneurons Is Affected by AMPA Receptor Activation in Slice Cultures of Embryonic Mouse Neocortex

description: It has been reported that functional α-amino-3-hydroxy-5-methyl-isoxazolpro-prionic acid (AMPA) receptors permeable to calcium are already expressed by tangentially migrating prospective interneurons in the developing rodent cerebral cortex. However, the role of these receptors is not yet fully understood. To examine the effect of activation of AMPA receptors on tangential migration, we exposed migrating prospective interneurons derived from the medial ganglionic eminence (MGE) to AMPA in slice cultures and performed time lapse imaging. In the neocortex, a subpopulation of MGE-derived cells stopped migration or changed the direction of migration in response to AMPA in a dose-dependent manner. In contrast, neither MGE-derived cells migrating in the subcortical territory nor radially migrating cells in the neocortex were affected by exposure to AMPA. Transfection of dominant-negative AMPA receptor subunit GluR1 to the tangentially migrating cells prevented the effects of AMPA on migration. This study provides evidence that the activation of AMPA receptors can directly affect tangential migration in the neocortical part of telencephalic slice cultures.
DOI: 10.6084/m9.figshare.5104678.v1
title: Supplementary Material for: Nocodazole-Induced Changes in Microtubule Dynamics Impair the Morphology and Directionality of Migrating Medial Ganglionic Eminence Cells

description: We have shown previously that actomyosin contractility plays an important role in controlling nuclear movements in future interneurons born in the medial ganglionic eminence (MGE) [Bellion et al.: J Neurosci 2005;25:5691–5699]. Because microtubules are known to control the structural and motile properties of migrating neurons, we asked whether alterations in the dynamic instability of microtubules would impair MGE cell migration. Migration was analyzed in flat cocultures in which green-fluorescent-protein-expressing MGE cells migrate on cortical cells from their explant of origin. A low (100 n<i>M</i>) concentration of nocodazole shortened the leading process of MGE cells that nevertheless continued to migrate at the same rate but frequently changed their direction of migration relative to control cells. MGE cells treated with a higher (1 µ<i>M</i>) concentration of nocodazole that strongly destabilized microtubules took on multipolar morphology. They extended thin and labile processes. MGE cells no longer exhibited directional migration and migration velocity slowed 2-fold. These results suggest that microtubule stability is crucial for maintaining polarity and controlling the directional migration of MGE cells, whereas additional mechanisms are required to control cell motility.
DOI: 10.6084/m9.figshare.5104657
title: Supplementary Material for: Extending the Platinum-Free Interval with a Non-Platinum Therapy in Platinum-Sensitive Recurrent Ovarian Cancer

description: <i>Background:</i> It has been proposed that extending the platinum-free interval with intervening non-platinum therapy increases the efficacy of a later re-treatment with platinum in platinum-sensitive recurrent ovarian cancer. This hypothesis is based on data from small series and although it has not been validated prospectively, this strategy has entered general practice in Italy in the last years. The SOCRATES study retrospectively assessed the pattern of care of a cohort of patients with recurrent platinum-sensitive ovarian cancer observed in the years 2000–2002 in 37 Italian centres. Data were collected between April and September 2005. <i>Methods:</i> Patients with recurrent ovarian cancer with a platinum-free interval &gt;6 months were eligible. 493 patient files were collected and 428 were eligible and analyzed. <i>Results:</i> The interval from the end of the 1st line to relapse was 6–12 months in 164 patients (39.5%) and &gt;12 months in 251 cases (60.5%). Patients received a 2nd (100%), 3rd (80.1%), 4th (50.2%), 5th (28.3%), and 6th (11.9%) line of chemotherapy. At 2nd line 282 (65.9%) received platinum (group A), while 146 (34.1%) received non-platinum chemotherapy (group B). In the latter group, 67 patients received platinum at later progression (group B1), while 79 never received platinum (group B2). Median time to platinum re-treatment was 20 and 23.1 months in patients of groups A and B1, respectively. The response rate to the first platinum received was 74.4 and 57.4% in groups A and B1, respectively (p = 0.02). Group B2 was characterized by the worst response rate and survival. At multivariate analysis time of first platinum re-treatment (2nd line vs. later; p = 0.0132; OR = 2.34) and age (p = 0.0029; OR = 2.41) was independently associated with a higher possibility of response to platinum. <i>Conclusions:</i> With the limits of a retrospective study, our data question the hypothesis that extending the platinum-free interval with an intervening non-platinum therapy in patients with recurrent platinum-sensitive ovarian cancer improves the response rate of a further platinum re-treatment.