Differential phosphorylation of some proteins of the neuronal cytoskeleton during brain development.

The cytoskeleton is important for neuronal morphogenesis. During the postnatal development of cat brain, the molecular composition of the neuronal cytoskeleton changes with maturation. Several of its proteins change in their rate of expression, in their degree of phosphorylation, in their subcellular distribution, or in their biochemical properties. It is proposed that phosphorylation is an essential mechanism to regulate the plasticity of the early, juvenile-type cytoskeleton. Among such proteins are several microtubule-associated proteins (MAPs), such as MAP5a, MAP2c or the juvenile tau proteins. Phosphorylation may also act on neurofilaments, postulated to be involved in the adult-type stabilization of axons. These observations imply that phosphorylation may affect cytoskeleton function in axons and dendrites at various developmental stages. Yet, the mechanisms of phosphorylation and its regulation cascades are largely unknown. In view of the topic of this issue on CD15, the potential role of matrix molecules being involved in the modulation of phosphorylation activity and of cytoskeletal properties is addressed.

Differential control of Notch1 gene transcription by Klf4 and Sp3 transcription factors in normal versus cancer-derived keratinocytes.

In specific cell types like keratinocytes, Notch signaling plays an important pro-differentiation and tumor suppressing function, with down-modulation of the Notch1 gene being associated with cancer development. Besides being controlled by p53, little else is known on regulation of Notch1 gene expression in this context. We report here that transcription of this gene is driven by a TATA-less "sharp peak" promoter and that the minimal functional region of this promoter, which extends from the -342 bp position to the initiation codon, is differentially active in normal versus cancer cells. This GC rich region lacks p53 binding sites, but binds Klf4 and Sp3. This finding is likely to be of biological significance, as Klf4 and, to a lesser extent, Sp3 are up-regulated in a number of cancer cells where Notch1 expression is down-modulated, and Klf4 over-expression in normal cells is sufficient to down-modulate Notch1 gene transcription. The combined knock-down of Klf4 and Sp3 was necessary for the reverse effect of increasing Notch1 transcription, consistent with the two factors exerting an overlapping repressor function through their binding to the Notch1 promoter.

Silver space-time trellis-coded modulation

Silver Code (SilC) was originally discovered in [1–4] for 2×2 multiple-input multiple-output (MIMO) transmission. It has non-vanishing minimum determinant 1/7, slightly lower than Golden code, but is fast-decodable, i.e., it allows reduced-complexity maximum likelihood decoding [5–7]. In this paper, we present a multidimensional trellis-coded modulation scheme for MIMO systems [11] based on set partitioning of the Silver Code, named Silver Space-Time Trellis Coded Modulation (SST-TCM). This lattice set partitioning is designed specifically to increase the minimum determinant. The branches of the outer trellis code are labeled with these partitions. Viterbi algorithm is applied for trellis decoding, while the branch metrics are computed by using a sphere-decoding algorithm. It is shown that the proposed SST-TCM performs very closely to the Golden Space-Time Trellis Coded Modulation (GST-TCM) scheme, yetwith a much reduced decoding complexity thanks to its fast-decoding property.

Modulation of angiogenic and inflammatory response in glioblastoma by hypoxia.

Glioblastoma are rapidly proliferating brain tumors in which hypoxia is readily recognizable, as indicated by focal or extensive necrosis and vascular proliferation, two independent diagnostic criteria for glioblastoma. Gene expression profiling of glioblastoma revealed a gene expression signature associated with hypoxia-regulated genes. The correlated gene set emerging from unsupervised analysis comprised known hypoxia-inducible genes involved in angiogenesis and inflammation such as VEGF and BIRC3, respectively. The relationship between hypoxia-modulated angiogenic genes and inflammatory genes was associated with outcome in our cohort of glioblastoma patients treated within prospective clinical trials of combined chemoradiotherapy. The hypoxia regulation of several new genes comprised in this cluster including ZNF395, TNFAIP3, and TREM1 was experimentally confirmed in glioma cell lines and primary monocytes exposed to hypoxia in vitro. Interestingly, the cluster seems to characterize differential response of tumor cells, stromal cells and the macrophage/microglia compartment to hypoxic conditions. Most genes classically associated with the inflammatory compartment are part of the NF-kappaB signaling pathway including TNFAIP3 and BIRC3 that have been shown to be involved in resistance to chemotherapy.Our results associate hypoxia-driven tumor response with inflammation in glioblastoma, hence underlining the importance of tumor-host interaction involving the inflammatory compartment.

Insulin modulation of luteinizing hormone secretion in normal female volunteers and lean polycystic ovary syndrome patients.

BACKGROUND/AIMS: Endocrine features of polycystic ovary syndrome (PCOS) include altered ovarian steroidogenesis, hyperinsulinemia and abnormal luteinizing hormone (LH) secretion. This study was undertaken to further evaluate the role of insulin to modulate LH secretion in lean PCOS patients with normal insulin sensitivity and normal volunteers. METHODS: The study was performed in five nonobese patients diagnosed with PCOS on the basis of amenorrhea and a polycystic morphology at ovarian ultrasound, and 5 normal controls in early to mid-follicular phase and matched for weight and age. All subjects were phenotyped, and then admitted for 12 h of frequent (q 10') blood sampling on two separate occasions, once for a baseline study and the other time for a hyperinsulinemic and euglycemic clamp study. LH was measured in samples obtained throughout each admission in order to perform LH pulse analysis. RESULTS: Baseline LH secretion in PCOS subjects was significantly different from controls: they had higher LH levels, higher LH/FSH ratios as well as a faster LH pulse frequency than normal women. Insulin administration did not affect the pattern of LH secretion of PCOS patients, whereas it significantly increased the LH pulse frequency while decreasing the LH interpulse intervals in the controls. CONCLUSIONS: These data confirm that an abnormal pattern of LH secretion characteristic of PCOS can be observed in lean patients, and appears independent of peripheral insulin levels. Furthermore, our results in lean controls provide the first direct evidence that peripheral insulin can modulate the activity of hypothalamic gonadotropin-releasing hormone (GnRH) neurons in the human.

Genotype-phenotype interactions in primary dystonias revealed by differential changes in brain structure.

Our understanding of how genotype determines phenotype in primary dystonia is limited. Familial young-onset primary dystonia is commonly due to the DYT1 gene mutation. A critical question, given the 30% penetrance of clinical symptoms in DYT1 mutation carriers, is why the same genotype leads to differential clinical expression and whether non-DYT1 adult-onset primary dystonia, with and without family history share pathophysiological mechanisms with DYT1 dystonia. This study examines the relationship between dystonic phenotype and the DYT1 gene mutation by monitoring whole-brain structure using voxel-based morphometry. We acquired magnetic resonance imaging data of symptomatic and asymptomatic DYT1 mutation carriers, of non-DYT1 primary dystonia patients, with and without family history and control subjects with normal DYT1 alleles. By crossing the factors genotype and phenotype we demonstrate a significant interaction in terms of brain anatomy confined to the basal ganglia bilaterally. The explanation for this effect differs according to both gene and dystonia status: non-DYT1 adult-onset dystonia patients and asymptomatic DYT1 carriers have significantly larger basal ganglia compared to healthy subjects and symptomatic DYT1 mutation carriers. There is a significant negative correlation between severity of dystonia and basal ganglia size in DYT1 mutation carriers. We propose that differential pathophysiological and compensatory mechanisms lead to brain structure changes in non-DYT1 primary adult-onset dystonias and DYT1 gene carriers. Given the range of age of onset, there may be differential genetic modulation of brain development that in turn determines clinical expression. Alternatively, a DYT1 gene dependent primary defect of motor circuit development may lead to stress-induced remodelling of the basal ganglia and hence dystonia.

Fast and slow voltage modulation of apical Cl- permeability in toad skin at high [K+]

The influence of voltage on the conductance of toad skin was studied to identify the time course of the activation/deactivation dynamics of voltage-dependent Cl- channels located in the apical membrane of mitochondrion-rich cells in this tissue. Positive apical voltage induced an important conductance inhibition which took a few seconds to fully develop and was instantaneously released by pulse inversion to negative voltage, indicating a short-duration memory of the inhibiting factors. Sinusoidal stimulation at 23.4 mM [Cl-] showed hysteresis in the current versus voltage curves, even at very low frequency, suggesting that the rate of voltage application was also relevant for the inhibition/releasing effect to develop. We conclude that the voltage modulation of apical Cl- permeability is essentially a fast process and the apparent slow components of activation/deactivation obtained in the whole skin are a consequence of a gradual voltage build-up across the apical membrane due to voltage sharing between apical and basolateral membranes

Gap junctions in isolated rat aorta: evidence for contractile responses that exhibit a differential dependence on intercellular communication

Connexin43 (Cx43) is a major gap junction protein present in the Fischer-344 rat aorta. Previous studies have identified conditions under which selective disruption of intercellular communication with heptanol caused a significant, readily reversible and time-dependent diminution in the magnitude of a1-adrenergic contractions in isolated rat aorta. These observations have indentified a significant role for gap junctions in modulating vascular smooth muscle tone. The goal of these steady-state studies was to utilize isolated rat aortic rings to further evaluate the contribution of intercellular junctions to contractions elicited by cellular activation in response to several other vascular spasmogens. The effects of heptanol were examined (0.2-2.0 mM) on equivalent submaximal (»75% of the phenylephrine maximum) aortic contractions elicited by 5-hydroxytryptamine (5-HT; 1-2 µM), prostaglandin F2a (PGF2a; 1 µM) and endothelin-1 (ET-1; 20 nM). Statistical analysis revealed that 200 µM and 500 µM heptanol diminished the maximal amplitude of the steady-state contractile responses for 5-HT from a control response of 75 ± 6% (N = 26 rings) to 57 ± 7% (N = 26 rings) and 34.9 ± 6% (N = 13 rings), respectively (P<0.05), and for PGF2a from a control response of 75 ± 10% (N = 16 rings) to 52 ± 8% (N = 19 rings) and 25.9 ± 6% (N = 18 rings), respectively (P<0.05). In contrast, 200 µM and 500 µM heptanol had no detectable effect on the magnitude of ET-1-induced contractile responses, which were 76 ± 5.0% for the control response (N = 38 rings), 59 ± 6.0% in the presence of 200 µM heptanol (N = 17 rings), and 70 ± 6.0% in the presence of 500 µM heptanol (N = 23 rings) (P<0.13). Increasing the heptanol concentration to 1 mM was associated with a significant decrease in the magnitude of the steady-state ET-1-induced contractile response to 32 ± 5% (21 rings; P<0.01); further increasing the heptanol concentration to 2 mM had no additional effect. In rat aorta then, junctional modulation of tissue contractility appears to be agonist-dependent.

The attentional modulation of the flash-lag effect

If a dot is flashed in perfect alignment with a pair of dots rotating around the visual fixation point, most observers perceive the rotating dots as being ahead of the flashing dot (flash-lag effect). This perceptual effect has been interpreted to result from the perceptual extrapolation of the moving dots, the differential visual latencies between flashing and moving stimuli, as well as the modulation of attentional mechanisms. Here we attempted to uncouple the attentional effects brought about by the spatial predictability of the flashing dot from the sensory effects dependent on its visual eccentricity. The stimulus was a pair of dots rotating clockwise around the fixation point. Another dot was flashed at either the upper right or the lower left of the visual field according to three separate blocked situations: fixed, alternate and random positions. Twenty-four participants had to judge, in all three situations, the location of the rotating dots in relation to the imaginary line connecting the flashing dot and the fixation point at the moment the dot was flashed. The flash-lag effect was observed in all three situations, and a clear influence of the spatial predictability of the flashing dot on the magnitude of the perceptual phenomenon was revealed, independently of sensory effects related to the eccentricity of the stimulus in the visual field. These findings are consistent with our proposal that, in addition to sensory factors, the attentional set modulates the magnitude of the differential latencies that give rise to the flash-lag phenomenon.

In vitro modulation of alkaline phosphatase activity of Saccharomyces cerevisiae grown in low or high phosphate medium

Our objective was to characterize the modulation of the activity of Saccharomyces cerevisiae alkaline phosphatases (ALPs) by classic inhibitors of ALP activity, cholesterol and steroid hormones, in order to identify catalytic similarities between yeast and mammalian ALPs. S. cerevisiae expresses two ALPs, coded for by the PHO8 and PHO13 genes. The product of the PHO8 gene is repressible by Pi in the medium. ALP activity from yeast (grown in low or high phosphate medium) homogenates was determined with p-nitrophenylphosphate as substrate, pH 10.4 (lPiALP or hPiALP, respectively). Activation of hPiALP was observed with 5 mM L-amino acids (L-homoarginine _ 186%, L-leucine _ 155% and L-phenylalanine - 168%) and with 1 mM levamisole (122%; percentage values, in comparison to control, of recovered activity). EDTA (5 mM) and vanadate (1 mM) distinctly inhibited hPiALP (2 and 20%, respectively). L-homoarginine (5 mM) had a lower activating effect on lPiALP (166%) and was the strongest hPiALP activator. Corticosterone (5 mM) inhibited hPiALP to 90%, but no effect was observed in low phosphate medium. Cholesterol, ß-estradiol and progesterone also had different effects on lPiALP and hPiALP. A concentration-dependent activation of lPiALP minus hPiALP was evident with all three compounds, most especially with ß-estradiol and cholesterol. These results do not allow us to identify similarities of the behavior of S. cerevisiae ALPs and any of the mammalian ALPs but allow us to raise the hypothesis of differential regulation of S. cerevisiae ALPs by L-homoarginine, ß-estradiol and cholesterol and of using these compounds to discriminate between S. cerevisiae lPiALP and hPiALP.

Glutamate-N-methyl-D-aspartate receptor modulation and minocycline for the treatment of patients with schizophrenia: an update

Growing consistent evidence indicates that hypofunction of N-methyl-D-aspartate (NMDA) transmission plays a pivotal role in the neuropathophysiology of schizophrenia. Hence, drugs which modulate NMDA neurotransmission are promising approaches to the treatment of schizophrenia. The aim of this article is to review clinical trials with novel compounds acting on the NMDA receptor (NMDA-R). This review also includes a discussion and translation of neuroscience into schizophrenia therapeutics. Although the precise mechanism of action of minocycline in the brain remains unclear, there is evidence that it blocks the neurotoxicity of NMDA antagonists and may exert a differential effect on NMDA signaling pathways. We, therefore, hypothesize that the effects of minocycline on the brain may be partially modulated by the NMDA-R or related mechanisms. Thus, we have included a review of minocycline neuroscience. The search was performed in the PubMed, Web of Science, SciELO, and Lilacs databases. The results of glycine and D-cycloserine trials were conflicting regarding effectiveness on the negative and cognitive symptoms of schizophrenia. D-serine and D-alanine showed a potential effect on negative symptoms and on cognitive deficits. Sarcosine data indicated a considerable improvement as adjunctive therapy. Finally, minocycline add-on treatment appears to be effective on a broad range of psychopathology in patients with schizophrenia. The differential modulation of NMDA-R neurosystems, in particular synaptic versus extrasynaptic NMDA-R activation and specific subtypes of NMDA-R, may be the key mediators of neurogenesis and neuroprotection. Thus, psychotropics modulating NMDA-R neurotransmission may represent future monotherapy or add-on treatment strategies in the treatment of schizophrenia.

Modulation différentielle par la privation de sommeil des processus attentionnels frontaux et pariétaux: une étude de potentiels évoqués cognitifs

L’objectif de la présente étude visait à évaluer les effets différentiels de la privation de sommeil (PS) sur le fonctionnement cognitif sous-tendu par les substrats cérébraux distincts, impliqués dans le réseau fronto-pariétal attentionnel, lors de l’administration d’une tâche simple et de courte durée. Les potentiels évoqués cognitifs, avec sites d’enregistrement multiples, ont été prévilégiés afin d’apprécier les effets de la PS sur l’activité cognitive rapide et ses corrélats topographiques. Le matin suivant une PS totale d’une durée de 24 heures et suivant une nuit de sommeil normale, vingt participants ont exécuté une tâche oddball visuelle à 3 stimuli. L’amplitude et la latence ont été analysées pour la P200 et la N200 à titre d’indices frontaux, tandis que la P300 a été analysée, à titre de composante à contribution à la fois frontale et pariétale. Suite à la PS, une augmentation non spécifique de l’amplitude de la P200 frontale à l’hémisphère gauche, ainsi qu’une perte de latéralisation spécifique à la présentation des stimuli cibles, ont été observées. À l’opposé, l’amplitude de la P300 était réduite de façon prédominante dans la région pariétale pour les stimuli cibles. Enfin, un délai de latence non spécifique pour la N200 et la P300, ainsi qu’une atteinte de la performance (temps de réaction ralentis et nombre d’erreurs plus élevé) ont également été objectivées. Les résultats confirment qu’une PS de durée modérée entraîne une altération des processus attentionnels pouvant être objectivée à la fois par les mesures comportementales et électrophysiologiques. Ces modifications sont présentes à toutes les étapes de traitement, tel que démontré par les effets touchant la P200, la N200 et la P300. Qui plus est, la PS affecte différemment les composantes à prédominance frontale et pariétale.

Expression des histones déméthylases dans les cellules hématopoïétiques humaines et les leucémies aiguës

L’importance des modificateurs de la chromatine dans la régulation de l’hématopoïèse et des hémopathies malignes est illustrée par l’histone méthyltransférase Mixed-Lineage Leukemia (MLL) qui est essentielle au maintien des cellules souches hématopoïétiques (CSH) et dont le gène correspondant, MLL, est réarrangé dans plus de 70% des leucémies du nourrisson. Les histones déméthylases (HDM), récemment découvertes, sont aussi impliquées dans le destin des CSH et des hémopathies malignes. Le but de ce projet est d’étudier l’expression des HDM dans les cellules hématopoïétiques normales et leucémiques afin d’identifier de potentiels régulateurs de leur destin. Nous avons réalisé un profil d'expression génique des HDM par qRT-PCR et par séquençage du transcriptome (RNA-seq) dans des cellules de sang de cordon (cellules CD34+ enrichies en CSH et cellules différenciées) et des cellules de leucémie aiguë myéloïde (LAM) avec réarrangement MLL. Les deux techniques montrent une expression différentielle des HDM entre les populations cellulaires. KDM5B et KDM1A sont surexprimés dans les cellules CD34+ par rapport aux cellules différenciées. De plus, KDM4A et PADI2 sont surexprimés dans les cellules leucémiques par rapport aux cellules normales. Des études fonctionnelles permettront de déterminer si la modulation de ces candidats peut être utilisée dans des stratégies d’expansion des CSH, ou comme cible thérapeutique anti-leucémique. Nous avons aussi développé et validé un nouveau test diagnostique pour détecter les mutations de GATA2 qui code pour un facteur de transcription clé de l’hématopoïèse impliqué dans les LAM. Ces travaux soulignent l’importance des facteurs nucléaires dans la régulation de l’hématopoïèse normale et leucémique.

Neurotensinergic modulation of glutamatergic neurotransmission in VTA neurons.

L’aire tegmentaire ventrale (VTA) contient une forte densité de terminaisons neurotensinergiques ainsi que des récepteurs à la surface des neurones dopaminergiques et non-dopaminergiques. Le VTA a été impliqué dans des maladies comme la schizophrénie, les psychoses et l’abus de substance. Les drogues d’abus sont connues pour induire le phénomène de sensibilisation - un processus de facilitation par lequel l’exposition à un stimulus produit une réponse augmentée lors de l’exposition subséquente au même stimulus. La sensibilisation se développe dans le VTA et implique mécanismes dopaminergiques et glutamatergiques. Il a été montré que les antagonistes neurotensinergiques bloquaient le développement de la sensibilisation et certains mécanismes de récompense et ces effets pourraient être médiés indirectement par une modulation de la neurotransmission glutamatergique. Cependant, on connaît peu les mécanismes de modulation de la transmission glutamatergique par la neurotensine (NT) dans le VTA. Le but de la présente thèse était d’étudier la modulation neurotensinergique de la neurotransmission glutamatergique dans les neurones dopaminergiques et non-dopaminergiques du VTA. Pour ce faire, nous avons utilisé la technique du patch clamp dans la cellule entière dans des tranches horizontales du VTA pour étudier les effets de différents agonistes et antagonistes neurotensinergiques. Les neurones ont été identifié comme Ih+ (présumés dopaminergiques) ou Ih- (présumés non-dopaminergiques) selon qu’ils exprimaient ou non un courant cationique activé par l’hyperpolarisation (Ih). Des techniques d’immunocytochimie ont été utilisées pour marquer les neurones et vérifier leur localisation dans le VTA. Dans une première étude nous avons trouvé que la neurotensine indigène (NT1-13) ou son fragment C-terminal, NT8-13, induisait une augmentation comparable des courants postsynaptiques excitateurs glutamatergiques (CPSEs) dans les neurones Ih+ ou Ih- du VTA. L'augmentation induite dans les neurones Ih+ par la NT8-13 a été bloquée par le SR48692, un antagoniste des récepteurs NTS1, et par le SR142948A, un antagoniste des récepteurs NTS1 et NTS2, suggérant que l'augmentation était médiée par l’activation des récepteurs NTS1. Dans les neurones Ih- l'augmentation n’a été bloquée que par le SR142948A indiquant une implication des récepteurs NTS2. Dans une deuxième étude, nous avons testé les effets de la D-Tyr[11]NT (un analogue neurotensinergique ayant différentes affinités de liaison pour les sous-types de récepteurs neurotensinergiques) sur les CPSEs glutamatergiques dans les neurones Ih+ et Ih- en parallèle avec une série d’expériences comportementales utilisant un paradigme de préférence de place conditionnée (PPC) menée dans le laboratoire de Pierre-Paul Rompré. Nous avons constaté que la D-Tyr[11]NT induisaient une inhibition dépendante de la dose dans les neurones Ih+ médiée par l'activation de récepteurs NTS2. En revanche, la D-Tyr[11]NT a produit une augmentation des CPSEs glutamatergiques médiée par des récepteurs NTS1 dans les neurones Ih-. Les résultats des expériences comportementales ont montré que des microinjections bilatérales de D-Tyr[11]NT dans le VTA induisait une PPC bloquée uniquement par la co-injection de SR142948A et SR48692, indiquant un rôle pour les deux types de récepteurs, NTS1 et NTS2. Cette étude nous a permis de conclure que i) la D-Tyr[11]NT agit dans le VTA via des récepteurs NTS1 et NTS2 pour induire un effet de récompense et ii) que cet effet est dû, au moins en partie, à une augmentation de la neurotransmission glutamatergique dans les neurones non-dopaminergiques (Ih-). Dans une troisième étude nous nous sommes intéressés aux effets de la D-Tyr[11]NT sur les réponses isolées médiées par les récepteurs N-méthyl-D-aspartate (NMDA) et acide α-amino-3- hydroxy-5-méthyl-4-isoxazolepropionique (AMPA) dans les neurones du VTA. Nous avons constaté que dans les neurones Ih+ l’amplitude des CPSEs NMDA et AMPA étaient atténuées de la même manière par la D-Tyr[11] NT. Cette modulation des réponses était médiée par les récepteurs NTS1 et NTS2. Au contraire, dans les neurones Ih-, l’amplitude des réponses NMDA et AMPA étaient augmentées en présence de D-Tyr[11]NT et ces effets dépendaient de l’activation des récepteurs NTS1 localisés sur les terminaisons glutamatergiques. Ces résultats fournissent une preuve supplémentaire que le NT exerce une modulation bidirectionnelle sur la neurotransmission glutamatergique dans les neurones du VTA et met en évidence un nouveau type de modulation peptidergique des neurones non-dopaminergiques qui pourrait être impliqué dans la sensibilisation. En conclusion, la modulation neurotensinergique de la neurotransmission glutamatergique dans les neurones dopaminergiques et non-dopaminergiques du VTA se fait en sens opposé soit, respectivement, par une inhibition ou par une excitation. De plus, ces effets sont médiés par différents types de récepteurs neurotensinergiques. En outre, nos études mettent en évidence une modulation peptidergique de la neurotransmission glutamatergique dans le VTA qui pourrait jouer un rôle important dans les mécanismes de lutte contre la toxicomanie.

Notes on analogue and digital amplitude modulation

Notes on AM, DSBSC, QAM, BPSK, 4QAM, 8PSK, 16QAM