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The identification of a critical role of tumour stroma in the regulation of tumour interstitial fluid pressure and the simultaneous discovery of the impact of anti-angiogenic drugs on tumour hemodynamics have provided new potential for improving tumour delivery of anti-cancer drugs. Here, we review the most recent studies investigating how tumour-associated fibroblasts and macrophages as well as the extracellular matrix itself may be targeted to facilitate delivery of both low-molecular weight drugs and macromolecules. In addition, we summarize the current understanding of the use of vasoactive compounds, radiotherapy and vascular-disrupting agents as potential adjuvants to maximize tumour delivery of anti-cancer drugs. The impact of these strategies on the diffusive and convective modes of drug transport is discussed in the light of Fick's and Starling's laws. Finally, we discuss how transcytosis through caveolae may also be exploited to optimize the selective delivery of conventional chemotherapy to the subendothelial tumour cell compartment.

Objectives We investigated whether retroinfusion of liposomal endothelial nitric oxide synthase (eNOS) S1177D complementary deoxyribonucleic acid (cDNA) would affect neovascularization and function of the ischemic myocardium. Background Recently, we demonstrated the feasibility of liposomal eNOS cDNA transfection via retroinfusion in a model of acute myocardial ischemia/reperfusion. In the present study, we used this approach to target a phosphomimetic eNOS construct (eNOS S1177D) into chronic ischemic myocardium in a pig model of hibernation. Methods Pigs (n = 6/group) were subjected to percutaneous implantation of a reduction stent graft into the left anterior descending artery (LAD), inducing total occlusion within 28 days. At day 28, retroinfusion of saline solution containing liposomal green fluorescent protein or eNOS S1177D cDNA (1.5 mg/animal, 2 x 10 min) was performed. Furthermore, L-nitroarginine-methylester (L-NAME) was applied orally from day 28, where indicated. At day 28 and day 49, fluorescent microspheres were injected into the left atrium for perfusion analysis. Regional functional reserve (at atrial pacing 140/min) was assessed at day 49 by subendocardial segment shortening (SES) (sonomicrometry, percent of ramus circumflexus region). Results The eNOS S1177D overexpression increased endothelial cell proliferation as well as capillary and collateral growth at day 49. Concomitantly, eNOS S1177D overexpression enhanced regional myocardial perfusion from 62 +/- 4% (control) to 77 +/- 3% of circumflex coronary artery-perfused myocardium, unless L-NAME was co-applied (69 +/- 5%). Similarly, eNOS S1177D cDNA improved functional reserve of the LAD (33 +/- 5% vs. 7 +/- 3% of circumflex coronary artery-perfused myocardium), except for L-NAME coapplication (13 +/- 6%). Conclusions Retroinfusion of eNOS S1177D cDNA induces neovascularization via endothelial cell proliferation and collateral growth. The resulting gain of perfusion enables an improved functional

Efficient behavior requires that internally specified motor plans be integrated with incoming sensory information. Motor preparation and visual signals converge in the intermediate and deep layers of the superior colliculus (SC) to influence saccade planning and execution; however, the mechanism by which these sometimes conflicting signals are combined remains unclear. We studied this issue by presenting visual distractors as monkeys prepared saccades toward an upcoming target whose timing and location were fully predictable. Monkeys made more distractor-directed errors when the spatial location of visual distractors more closely coincided with the saccadic goal. Concomitant pretarget activity of SC visuomotor neurons, whose response fields were centered on the saccadic goal, was similarly increased by the presentation of nearby distractors and inhibited by the presentation of distant distractors. Finally, subthreshold microstimulation of the SC shifted the pattern of distractor-directed errors away from the saccadic goal toward that specified by the site of stimulation. Together, our results suggest that the likelihood of saccade generation is influenced by the spatial register of internal motor preparation signals and external sensory signals across the topographically organized SC map.

Hypoxia-inducible factor 1 (HIF-1) is a master transcriptional factor. Under normal oxygen tension, HIF-1 activity is usually suppressed due to the rapid, oxygen-dependent degradation of one of its two subunits, HIF-1 alpha. Here we report that normoxic HIF-1 activity can be upregulated through NO-mediated S-nitrosylation and stabilization of HIF-1 alpha. In murine tumors, exposure to ionizing radiation stimulated the generation of NO in tumor-associated macrophages. As a result, the HIF-1 alpha protein is S-nitrosylated at Cys533 (through "biotin switch" assay) in the oxygen-dependent degradation domain, which prevents its destruction. Importantly, this mechanism appears to be independent of the prolylhydroxylase-based pathway that is involved in oxygen-dependent regulation of HIF-1 alpha. Selective disruption of this S-nitrosylation significantly attenuated both radiation-induced and macrophage-induced activation of HIF-1 alpha. This interaction between NO and HIF-1 sheds new light on their involvement in tumor response to treatment as well as mammalian inflammation process in general.

Recent studies have shown that same-race (SR) faces are processed more holistically than other-race (OR) faces, a difference that may underlie the greater difficulty at recognizing OR than SR faces (the "other-race effect"). This article provides original evidence suggesting that the holistic processing of faces may be sensitive to the observers' racial categorization of the face. In Experiment 1, Caucasian participants performed a face-composite task with Caucasian faces, Asian faces, and racially ambiguous morphed face stimuli. Identical morphed face stimuli were processed more holistically when categorized as SR than as OR faces. Experiment 2 further suggests that this finding was not underlain by strategic or training effects. Overall, these results support the view that one's categorization of a face as belonging to the same or another race plays a critical role in the holistic processing of this face.

Objectives: This article outlines the role of testosterone in prostate cancer (PCa) homeostasis and describes the effect of androgen-deprivation therapy (ADT) by luteinising hormone-releasing hormone (LHRH) agonists on PCa. In addition, the outcome and benefits of medical versus surgical castration are evaluated. Methods: This paper is based on a presentation during a satellite symposium held at the European Association of Urology 2007 annual meeting. Relevant articles on testosterone importance and hormonal control in PCa were selected from literature databases. Results: The goal of ADT is to induce a rapid, profound, and sustained suppression of testosterone. The testosterone threshold to trigger apoptosis is not yet known; therefore, it is hypothesised that it is better to achieve lower testosterone values. Historically, it has been assumed that ADT succeeds if testosterone levels reach a threshold of 50 ng/dl or lower. However, surgical castration, which remains the "gold standard", achieves lower testosterone values, closer to 20 ng/dl. Analysis of clinical trials with conventional LHRH agonists shows that these agents fail to reduce testosterone to <= 50 ng/dl in 1-12.5% of the patients, whereas 13-40% of the cases do not achieve testosterone levels <= 20 ng/dl. Furthermore, the incidence of miniflares to repeat injections varies around 10%, whereas breakthrough responses occur in 2-24% of the patients. Conclusions: Conventional LHRH agonists do not induce profound and sustained testosterone suppression in all patients, similar to orchidectomy. To give patients maximum benefit from medical castration via LHRH agonist treatment, we must have new formulations. (c) 2007 European Association of Urology. Published by Elsevier B.V. All rights reserved.

A significant proportion of patients with prostate cancer experience biochemical failure following primary therapy. Of these, some may be at high risk of dying from prostate cancer. Salvage therapy can improve survival outcomes in these patients but at the cost of adverse effects that may affect quality of life. The choice of salvage therapy depends on the location of the tumour recurrence and how aggressive the disease is. This article examines the treatment options for patients with a rising prostate-specific antigen (PSA) level after radical prostatectomy or radiotherapy, using two clinical scenarios. After radical prostatectomy fails, radiotherapy is a salvage option for low-risk patients with local disease recurrence; however, hormonal therapy, with or without radiotherapy, may represent a better option for patients with high-risk disease. Local salvage therapy options in patients with PSA failure after radiotherapy include brachytherapy, cryotherapy, high-intensity focused ultrasound, and surgery. In patients with moderate- to high-risk disease, most clinicians would also prescribe some form of hormonal treatment. (c) 2006 European Association of Urology. Published by Elsevier B.V. All rights reserved.

Objectives: This review paper discusses the importance of optimal testosterone control in prostate cancer (PCa) and the advantages of the luteinising hormone-releasing hormone (LHRH) agonist Eligard (R) in this respect. Methods: During an expert meeting in Marbella, Spain, in October 2006, PCa experts discussed optimal testosterone control and the role of Eligard. An interactive voting on these topics was held among the delegates. This paper reports on this meeting. Results: LHRH agonists should suppress testosterone levels to an extent equivalent to surgical castration. A significant proportion of patients on conventional LHRH agonists do not achieve surgical castration levels. In addition, a significant percentage of patients experience hormonal escape during treatment. Eligard uses a modern delivery system and offers 1-, 3- and 6-mo depot formulations. The Eligard 1-mo, 3-mo, and 6-mo formulations achieve testosterone levels <= 20 ng/dl in up to 98% of patients. Injection-related and breakthrough testosterone escapes seldom occur. Eligard 6-mo formulation (45 mg; Eligard 6) is the first LHRH agonist able to extend treatment for 6 mo. Eligard 6 allows flexible monitoring of patients because the need for visits solely to administer injections is removed. In addition, reducing the number of injections may help reduce psychological burden in patients who then are less often reminded of their condition. Conclusions: Eligard provides optimal testosterone control in PCa patients, that is, achieves castration testosterone levels and is associated with a very low percentage of hormonal escapes. Eligard 6 offers flexibility in follow-up of patients and may help reduce emotional distress. (c) 2007 European Association of Urology. Published by Elsevier B.V. All rights reserved.

Bilateral orchiectomy became the "gold standard" treatment option for advanced prostate cancer following the Nobel Prize winning study by Huggins and Hodges in 1941. Almost 40 yr later, the combination of a nonsteroidal antiandrogen with castration-based treatment (combined androgen blockade [CAB]) was proposed to offer superior response and survival compared with castration alone. However, 25 yr and many randomised studies later, the benefit of CAB remains controversial. Here, we present a clinical scenario that describes a patient with advanced prostate cancer and we review the available treatment options. The first part of the article focuses on androgen-dependent disease, comparing castration (surgical or medical) to either Casodex (TM) (bicalutamide) 150 mg monotherapy or CAB. In the CAB setting, Casodex 50 mg combined with castration-based therapy is calculated to reduce the rate of all-cause mortality by an estimated 20% compared with castration alone. The second part of the article describes treatment options for androgen-independent prostate cancer, including alternative forms of hormonal therapy, such as oestrogen therapy and ketoconazole- and docetaxel-based chemotherapy. (c) 2006 European Association of Urology. Published by Elsevier B.V. All rights reserved.

L'ophtalmopathie thyroïdienne est la manifestation extra-thyroïdienne la plus fréquente de la maladie de Basedow. L'approche multidisciplinaire de cette maladie permet la prise en charge globale de cette maladie invalidante. Aux Cliniques universitaires Saint-Luc à Bruxelles , les patients atteints de cette pathologie sont systématiquement pris en charge par «la clinique de l’orbitopathie thyroïdienne». Cet article résume les aspects endocriniens et ophtalmologiques de cette maladie, ainsi que des recommandations pratiques pour les généralistes et les spécialistes

Interhemispheric inhibition (IHI) between motor cortical areas is thought to play a critical role in motor control and could influence manual dexterity. The purpose of this study was to investigate IHI preceding movements of the dominant and nondominant hands of healthy volunteers. Movement-related IHI was studied by means of a double-pulse transcranial magnetic stimulation protocol in right-handed individuals in a simple reaction time paradigm. IHI targeting the motor cortex contralateral (IHI(c)) and ipsilateral (IHI(i)) to each moving finger was determined. IHI(c) was comparable after the go signal, a long time preceding movement onset, in both hands. Closer to movement onset, IHI(c) reversed into facilitation for the right dominant hand but remained inhibitory for left nondominant hand movements. IHI(i) displayed a nearly constant inhibition with a trough early in the premovement period in both hands. In conclusion, our results unveil a more important modulation of interhemispheric interactions during generation of dominant than nondominant hand movements. This modulation essentially consisted of a shift from a balanced IHI at rest to an IHI predominantly directed toward the ipsilateral primary motor cortex at movement onset. Such a mechanism might release muscles from inhibition in the contralateral primary motor cortex while preventing the occurrence of the mirror activity in ipsilateral primary motor cortex and could therefore contribute to intermanual differences in dexterity.

Numerous studies report a negative event-related potential at occipito-temporal scalp sites between 130 and 200 ms (N170) that is larger when elicited by faces than by other object categories. Thierry and colleagues argued that this effect reflects an artifact of uncontrolled interstimulus perceptual (physical) variance (ISPV), which when controlled eliminates the difference between faces and nonfaces.

Although the molecular function of sigma receptors has not been fully defined and the natural ligand(s) is still not known, there is increasing evidence that these receptors and their ligands might play a significant role in cancer biology. 4-(N-benzylpiperidin-4-yl)-4-iodobenzamide (4-IBP), a selective sigma1 agonist, has been used to investigate whether this compound is able to modify: 1) in vitro the migration and proliferation of human cancer cells; 2) in vitro the sensitivity of human glioblastoma cells to cytotoxic drugs; and 3) in vivo in orthotopic glioblastoma and non-small cell lung carcinoma (NSCLC) models the survival of mice co-administered cytotoxic agents. 4-IBP has revealed weak antiproliferative effects on human U373-MG glioblastoma and C32 melanoma cells but induced marked concentration-dependent decreases in the growth of human A549 NSCLC and PC3 prostate cancer cells. The compound was also significantly antimigratory in all four cancer cell lines. This may result, at least in U373-MG cells, from modifications to the actin cytoskeleton. 4-IBP modified the sensitivity of U373-MG cells in vitro to proapoptotic lomustin and proautophagic temozolomide, and markedly decreased the expression of two proteins involved in drug resistance: glucosylceramide synthase and Rho guanine nucleotide dissociation inhibitor. In vivo, 4-IBP increased the antitumor effects of temozolomide and irinotecan in immunodeficient mice that were orthotopically grafted with invasive cancer cells.

While the neurotensin (NT) roles in pancreatic cancer growth are well documented, its effects on pancreatic cancer cell migration have not been described.

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Background and Aims: We have identified six adipokines that are oversecreted by omental adipose tissue (OAT) in obesity: 3 chemokines [Growth-Related Oncogen factor (GRO), Regulated upon Activation Normal T cells Expressed and Secreted (RANTES), Macrophage Inflammatory Protein-1ß (MIP-1ß)], 1 interleukin (IL-7), 1 tissue inhibitor of metalloproteinases (TIMP-1) and 1 growth factor (thrombopoietin, TPO). Enhanced expression of these adipokines in adipocytes did correlate with several features of the metabolic syndrome. The aim of the present study was to examine whether the abnormal hormonal milieu and/or the chronic inflammation state that prevail in obesity could contribute to such dysregulation of adipokine production. We thus examined the effect of some hormones (insulin, glucocorticoids and catecholamines) and the proinflammatory cytokine, TNFĮ on these adipokines in omental adipocytes.[...]

Large, rapid gaze shifts necessitate intricate coordination of the eyes and head. Brief high-frequency bursts of activity within the intermediate and deeper layers of the superior colliculus (dSC) encode desired gaze shifts regardless of component movements of the eyes and head. However, it remains unclear whether low-frequency activity emitted by oculomotor neurons within the dSC and elsewhere has any role in eye-head gaze shifts. Here we test the hypothesis that such low-frequency activity contributes to eye-head coordination by selectively priming head premotor circuits. We exploited the capacity for short-duration (10 ms, 4 pulses) dSC stimulation to evoke neck muscle responses without compromising ocular stability, stimulating at various intervals of a "gap-saccade" task. Low-frequency neural activity in many oculomotor areas (including the dSC) is known to increase during the progression of the gap-saccade task. Stimulation was passed during either a fixation-interval while a central fixation point was illuminated, a 200-ms gap-interval between fixation point offset and target onset, or a movement-interval following target onset. In the two monkeys studied, the amplitude of evoked responses on multiple neck muscles tracked the known increases in low-frequency oculomotor activity during the gap-saccade task, being greater following stimulation passed at the end of the gap- versus the fixation-interval, and greater still when the location of stimulation during the movement interval coincided with the area of the dSC generating the ensuing saccade. In one of these monkeys, we obtained a more detailed timeline of how these results co-varied with low-frequency oculomotor activity by stimulating, across multiple trials, at different times within the fixation-, gap- and movement-intervals. Importantly, in both monkeys, baseline levels of neck EMG taken immediately prior to stimulation onset did not co-vary with the known pattern of low-frequency oculomotor activity u

The existence of a possible common mechanism for duration and numerosity processing was tested with a Stroop task. Participants had to compare either the duration or the numerosity of sequences of flashing dots for which the duration and numerosity were independently manipulated to create congruent, incongruent or neutral pairs. Results show that the numerical cues interfered with duration processing, whereas the temporal cues did not interfere with numerosity processing. These findings extend the idea of an automatic access to magnitude to non-symbolic sequentially presented material, and reflect a probable difference in the mandatory processing of numerosity and duration.

Aminoadamantanes are commonly used in the treatment of Parkinson's and Alzheimer's diseases. While these drugs are shown to antagonise ionotropic glutamate receptors on neuronal cells, additional mechanisms could contribute to their neuroprotective properties. The aim of the present study was to investigate the effect of aminoadamantanes on the production of the glial cell line-derived neurotrophic factor (GDNF) in glial cells. For this purpose, we measured the modulation of GDNF release in C6 glioma cell cultures treated for 24 h with amantadine and memantine. Both drugs dose-dependently increased GDNF level in the culture medium with similar potency (submicromolar range) and efficacy (three to four-fold induction). RT-PCR studies revealed that both compounds also increased GDNF mRNA levels and their influence on the GDNF gene transcription was further evidenced using a rat GDNF promoter luciferase reporter assay. Together, these results demonstrate that the neuroprotective effect of amantadine and memantine could involve the regulation of GDNF production by glial cells.

We previously showed that the stimulation of heme oxygenase-1 expression by high glucose and hydrogen peroxide (H(2)O(2)) in cultured rat islets is prevented by antioxidants and suggested that this effect of high glucose results from an oxidative stress. However, the role of oxidative stress in high-glucose-induced beta-cell dysfunction is unclear. We therefore compared the preventative effects of N-acetyl-l-cysteine (NAC), a free radical scavenger, and manganese(III)tetrakis (4-benzoic acid)porphyrin (MnTBAP), a superoxide dismutase/catalase mimetic agent, on the alteration of stimulus-secretion coupling induced in rat islets by overnight exposure to hydrogen peroxide (H(2)O(2)-treated islets) or 1-wk culture in 30 vs. 10 mmol/l glucose (High-glucose vs. Control islets). The features of beta-cell dysfunction differed between the two groups: reduced glucose-induced insulin secretion without changes in glucose sensitivity in H(2)O(2)-treated islets; increased sensitivity to glucose with parallel reductions in insulin content and maximal rate of glucose-induced insulin secretion in High-glucose islets. The latter alterations were accompanied by a decrease in preproinsulin without changes in pancreatic and duodenal homeobox gene 1 mRNA levels. The functional alterations induced by H(2)O(2) were significantly prevented by addition of NAC or MnTBAP in the culture medium. In contrast, neither NAC nor MnTBAP affected the functional alterations induced by high glucose. These results suggest that beta-cell dysfunction induced by 1-wk culture in high glucose does not result from an increase in oxidative stress.

Spatial attention paradigms using auditory or tactile stimulation were used to explore neural and behavioral reorganization in early blind subjects. Although it is commonly assumed that blind subjects outperform sighted subjects in such tasks, the empirical data to confirm this remain controversial. Moreover, previous studies have often confounded factors of sensory acuity with those of attention. In the present work, we compared the performance of individually matched early blind and sighted subjects during auditory and tactile tasks. These consisted of sensory acuity tests, simple reaction time task as well as selective and divided spatial attention tasks. Based on sensory measurements, we made sure that the reliability and salience of auditory and tactile information were identical between the two populations to estimate attentional performance independently of sensory influence. Results showed no difference between groups in either sensory sensitivity or simple reaction time task for both modalities. However, blind subjects displayed shorter reaction times than sighted subjects in both tactile and auditory selective spatial attention tasks and also in bimodal divided spatial attention tasks. The present study thus demonstrates an enhanced attentional performance in early blind subjects which is independent of sensory influence. These supra-normal abilities could be related to quantitative and qualitative changes in the way early visually deprived subjects process non-visual spatial information.

Contrasting with its robust expression during embryogenesis, the glial cell line-derived neurotrophic factor (GDNF) is repressed in the adult organism. However, rapid induction of this neuronal growth factor is observed following diverse neuronal insults and it is now widely accepted that the control of its expression could constitute a powerful target in neuropharmacology. We investigated the effects of the neuroprotective drug, riluzole, on the GDNF gene expression in glial cells. Exposure of C6 glioma cells to riluzole (1 microM) significantly increased GDNF protein and mRNA levels. Using luciferase reporter gene constructs encoding fragments of the 5' untranslated region of the rat GDNF gene, we demonstrated that riluzole mediates its effect at the transcription level. Furthermore, luciferase assays revealed the presence of a negative regulatory region within the +343/+587 region of exon 1. This region was shown to contribute to the high sensitivity and specificity of the induction mediated by riluzole in the C6 glioma cell line at pharmacologically relevant concentrations. The effects of riluzole were inhibited by the mitogen-activated protein kinase extracellular signal-related kinase (MEK) inhibitor PD 98059. Together, these results indicated that the induction of GDNF release by riluzole in the C6 glioma cells results from the activation of its corresponding gene promoter through a signalling pathway involving MEK activity. This study suggests that the regulation of GDNF gene transcription in glial cells could contribute to the pharmacological properties of riluzole and possibly other neuroprotective drugs.

PURPOSE: This study involved a volunteer completely blind from retinis pigmentosa who had previously been implanted with an optic nerve visual prosthesis. The aim of this two-year study was to train the volunteer to localize a given object in nine different positions, to discriminate the object within a choice of six, and then to grasp it. METHODS: In a closed-loop protocol including a head worn video camera, the nerve was stimulated whenever a part of the processed image of the object being scrutinized matched the center of an elicitable phosphene. The accessible visual field included 109 phosphenes in a 14 degrees x 41 degrees area. RESULTS: Results showed that training was required to succeed in the localization and discrimination tasks, but practically no training was required for grasping the object. The volunteer was able to successfully complete all tasks after training. The volunteer systematically performed several left-right and bottom-up scanning movements during the discrimination task. Discrimination strategies included stimulation phases and no-stimulation phases of roughly similar duration. CONCLUSION: This study provides a step towards the practical use of the optic nerve visual prosthesis in current daily life.

Numerous difficulties arise during in vivo measurements of transepithelial nasal potential difference (PD) in mice, such as inadequate duration and depth of anaesthesia, bronchoaspiration of solutions perfused in the nose, and respiratory and/or cardiovascular depression. Anaesthesia was induced in adult C57 mice with intraperitoneal injection of a combination of fentanyl, droperidol and medetomidine, each of these at either a small dose (0.20, 10 and 0.33 mg/kg, respectively) or at a large dose (0.40, 20 and 0.40 mg/kg, respectively), combined with a fixed dose of 0.375 microg clonidine. In order to establish a pharmacokinetic-pharmacodynamic relationship, blood concentrations of the first three drugs were measured in 24 animals by liquid-chromatography tandem mass spectrometry. At the end of the experiment, naloxone, a competitive morphinic antagonist, and atipamezole, an alpha-2 adrenergic antagonist, were administered. Bronchoaspiration was prevented by tilting the animal head downwards and by absorbing the excess fluid from the opposite nostril and from the oral cavity. Optimal assessment of anaesthesia associated with regular respiration, loss of blink, pupillary and pedal withdrawal reflexes was obtained with doses of fentanyl, droperidol and medetomidine corresponding to 0.20, 20 and 0.40 mg/kg, respectively. Blood concentrations of fentanyl around 17 ng/mL induced loss of respiratory efforts and were followed by death during the experiment. Integrity of ion transport was demonstrated under continuous perfusion by successive depolarization after amiloride and repolarization after chloride-free solution. The combination investigated in this study lead to adequate surgical anaesthesia (stage III, plane 2) for prolonged nasal PD measurements in spontaneously breathing mice.

We have previously shown that some visual motion areas can be specifically recruited by auditory motion processing in blindfolded sighted subjects [Poirier, C., Collignon, O., De Volder, A.G., Renier, L., Vanlierde, A., Tranduy, D., Scheiber, C., 2005. Specific activation of V5 brain area by auditory motion processing: an fMRI study. Brain Res. Cogn. Brain Res. 25, 650-658]. The present fMRI study investigated whether auditory motion processing may recruit the same brain areas in early blind subjects. The task consisted of simultaneously determining both the nature of a sound stimulus (pure tone or complex sound) and the presence or absence of its movement. When a movement was present, blind subjects had to identify its direction. Auditory motion processing, as compared to static sound processing, activated the brain network of auditory and visual motion processing classically observed in sighted subjects. Accordingly, brain areas previously considered as specific to visual motion processing could be specifically recruited in blind people by motion stimuli presented through the auditory modality. This indicates that the occipital cortex of blind people could be organized in a modular way, as in sighted people. The similarity of these results with those we previously observed in sighted subjects suggests that occipital recruitment in blind people could be mediated by the same anatomical connections as in sighted subjects.

The mammalian homologues of the Drosophila transient receptor potential (TRP) represent a superfamily of ion channels involved in Ca(2+) homeostasis. Several members of this family are activated either by a depletion of the internal stores of Ca(2+) or by stimulation of G protein-coupled receptors. In androgen responsive prostate cancer cell line LNCaP, TRPC1, TRPC4 and/or TRPV6 have been reported to function as store-operated channels (SOCs) while TRPC3 might be involved in the response to agonist stimulation, possibly through the induction of diacylglycerol production by phospholipase C. However, the control of expression of these TRP proteins is largely unknown. In the present study, we have investigated if the expression of the TRP proteins possibly involved in the capacitative influx of calcium is influenced by the contents of Ca(2+) in the endoplasmic reticulum. Using real-time PCR and Western blot techniques, we show that the expression of TRPC1, TRPC3 and TRPV6 proteins increases after a prolonged (24-48 h) depletion of the stores with thapsigargin. The upregulation of TRPC1 and TRPC3 depends on the store contents level and involves the activation of the Ca(2+)/calmodulin/calcineurin/NFAT pathway. Functionally, cells overexpressing TRPC1, TRPC3 and TRPV6 channels after a prolonged depletion of the stores showed an increased [Ca(2+)](i) response to alpha-adrenergic stimulation. However, the store-operated entry of calcium was unchanged. The isolated overexpression of TRPV6 (without overexpression of TRPC1 and TRPC3) did not produce this increased response to agonists, therefore suggesting that TRPC1 and/or TRPC3 proteins are responsible for the response to alpha-adrenergic stimulation but that TRPC1, TPRC3 and TRPV6 proteins, expressed alone or concomitantly, are not sufficient for SOC formation.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a selective loss of motor neurones accompanied by intense gliosis in lesioned areas of the brain and spinal cord. Glutamate-mediated excitotoxicity resulting from impaired astroglial uptake constitutes one of the current pathophysiological hypotheses explaining the progression of the disease. In this study, we examined the regulation of glutamate transporters by type 5 metabotropic glutamate receptor (mGluR5) in activated astrocytes derived from transgenic rats carrying an ALS-related mutated human superoxide dismutase 1 (hSOD1(G93A)) transgene. Cells from transgenic animals and wild-type littermates showed similar expression of glutamate-aspartate transporter and glutamate transporter 1 (GLT-1) after in vitro activation, whereas cells carrying the hSOD1 mutation showed a three-fold higher expression of functional mGluR5, as observed in the spinal cord of end-stage animals. In cells from wild-type animals, (S)-3,5-dihydroxyphenylglycine (DHPG) caused an immediate protein kinase C (PKC)-dependent up-regulation of aspartate uptake that reflected the activation of GLT-1. Although this effect was mimicked in both cultures by direct activation of PKC using phorbol myristate acetate, DHPG failed to up-regulate aspartate uptake in cells derived from the transgenic rats. The failure of activated mGluR5 to increase glutamate uptake in astrocytes derived from this animal model of ALS supports the theory of glutamate excitotoxicity in the pathogenesis of the disease.

BACKGROUND: In the near future, adult porcine islets of Langerhans appear as an unlimited source of insulin-producing cells which could play a major role for treating diabetes mellitus. There is, however, an obvious lack of pre-clinical results and data in the pig-to-primate model. One of the main hurdles of this model is certainly related to the difficulty of reproducing regularly successful porcine islet isolation. This experimental work was designed to provide guidelines applicable in pig pancreas procurement and islet isolation for successful islet xenotransplantation into primates. METHODS: Pancreases were harvested from adult Belgium Landrace pigs (n = 79) in a single centre. The impact on islet yield of (1) pancreas procurement (blood exsanguination and warm ischaemia time (WIT)), (2) cold storage solutions (classic UW and modified UW (without hydroxyethyl starch and inverse K+/Na+ concentration)), (3) a dynamic or static method of pancreas digestion, and (4) the endotoxin content and enzymatic activity from five different batches of Liberase PI was studied. In addition, pancreatic biopsies (n = 18), performed before isolation, were retrospectively analyzed to study the impact of histomorphometry on porcine islet yield. Finally, two diabetic cynomolgus monkeys were transplanted without immunosuppression with 15,000 pig islet equivalents/kg body weight of recipient to assess in vivo the function of freshly isolated islets. Univariate and multivariate analyses were performed. RESULTS: By multiple linear regression, the most significant variables that significantly improved islet yield were, firstly, the presence of <30 EU (endotoxin units) of endotoxin in Liberase batches, followed by a WIT under 10 min and the use of blood exsanguination before pancreas harvesting (P < 0.005). In contrast, isolation method (dynamic vs. static) and the solution used for storage (short-term) (UW vs. modified UW) did not significantly influence islet yield. The correlation of ret

BACKGROUND/AIMS: The endothelium has been recognized as a key component in the regulation of blood vessels. We designed a simple procedure to separate endothelial and smooth muscle RNA from rat aorta and mesenteric artery and used this method to establish the distribution of Na(+)/K(+)-ATPase alpha-subunit isoforms (NaKalpha1, NaKalpha2 and NaKalpha3) within the arterial wall. METHODS: Rat aorta was perfused with Tripure, a reagent for RNA isolation, yielding 3 successive RNA fractions (E1-E3) and the remaining tissular RNA (Ao[E-]). A similar procedure was applied to the mesenteric artery. Gene expression was studied by semiquantitative reverse-transcription polymerase chain reaction. RESULTS: Compared to unperfused aorta (Ao[E+]), typical endothelial mRNAs were enriched 3- to 5-fold in E1-E3 but almost absent in Ao[E-], whereas smooth muscle mRNAs were low in E1-E3 but similarly expressed in Ao[E-] and Ao[E+]. NaKalpha1 was uniformly expressed in all fractions, NaKalpha2 closely followed the expression pattern of smooth muscle markers and NaKalpha3 expression was weak and attributable to blood contamination. Comparable results were obtained with the mesenteric artery. CONCLUSION: We conclude that, in aorta and mesenteric artery, Tripure perfusion allows for a rapid and reliable separation of endothelial mRNA from smooth muscle mRNA, and that endothelium only expresses NaKalpha1, whereas smooth muscle expresses NaKalpha1 and NaKalpha2, but not NaKalpha3.

In skeletal muscle, Ca(2+) is implicated in contraction, and in regulation of gene expression. An alteration of [Ca(2+)](i) homeostasis is responsible, at least partially, for the muscle degeneration that occurs after eccentric contractions in Duchenne muscular dystrophy, a disease characterized by the loss of the cytoskeletal protein dystrophin. Using patch clamp in the cell-attached configuration, we characterized the store-operated channels (SOCs) and the stretch-activated channels (SACs) present in isolated mouse skeletal muscle. SOCs were voltage independent, had a unitary conductance between 7 and 8 pS (110 mm Ca(2+) in the pipette), and their open probability increased when the sarcoplasmic reticulum was depleted by thapsigargin. These SOCs were identical to those previously described in the pathophysiology of Duchenne muscular dystrophy. Under the same experimental conditions, we detected a channel activity that was increased by applying a negative pressure to the patch electrode. The SACs responsible for this current had the same unitary conductance and current-voltage relationship as those observed for SOCs. SOCs and SACs had a similar sensitivity to pharmacological agents such as Gd(3+), SKF-96365, 2-aminoethoxydiphenyl borate and GsMTx4 toxin. Moreover, stimulation with IGF-1 increased the occurrence of the activity of both channel types. Together, these observations suggest that SOCs and SACs might belong to the same population or share common constituents. From a functional point of view, treatment of soleus muscle with SKF-96365 or GsMTx4 toxin increased its sensitivity to a fatigue protocol, suggesting that the influx of Ca(2+) that occurs through these channels during contraction is also involved in force maintaining during repeated stimulations.

Research into visual prosthetics is expected to revolutionize the treatment of blind patients with incurable outer retinal degenerative disease. Substantial evidence shows that useful visual sensations can be produced by controlled electrical stimulation of the optic nerve. To make the optic nerve visual prosthesis more acceptable, implantation techniques safer and less invasive than those previously used have been developed. A medial transconjunctival approach is now used to implant a stimulating electrode around the intraorbital section of the optic nerve. This new technique allows sufficient exposure of the nerve after detaching only one rectus muscle and performing a lateral canthotomy. Previously, an electrode was implanted in the intracranial part of the optic nerve, which required more invasive surgery. The new technique was first developed in cadavers and in patients undergoing eye enucleations. Finally, a 68-year-old blind man suffering from retinitis pigmentosa underwent long-term implantation. In this case report the authors describe the technique and outline some of the challenges involved.

Two identical top parts of a face photograph look different if their bottom parts differ. This perceptual illusion, the "face composite effect", is taken as strong evidence that faces are processed as a whole rather than as a collection of independent features. To test the hypothesis that areas responding preferentially to faces in the human brain represent faces holistically, we recorded functional magnetic resonance imaging (fMRI) during an adaptation paradigm with the composite face illusion. In both the middle fusiform gyrus (MFG) and the inferior occipital gyrus (IOG), we observed a significantly larger response to the same top face when it was aligned with different bottom parts than with the same bottom part, with a most robust effect in the right middle fusiform gyrus. This difference was not found when the top and the bottom face parts were spatially misaligned or when the faces were presented upside-down. These findings indicate that facial features are integrated into holistic face representations in areas of the human visual cortex responding preferentially to faces.

Recognizing individual faces outside one's race poses difficulty, a phenomenon known as the other-race effect. Most researchers agree that this effect results from differential experience with same-race (SR) and other-race (OR) faces. However, the specific processes that develop with visual experience and underlie the other-race effect remain to be clarified. We tested whether the integration of facial features into a whole representation-holistic processing-was larger for SR than OR faces in Caucasians and Asians without life experience with OR faces. For both classes of participants, recognition of the upper half of a composite-face stimulus was more disrupted by the bottom half (the composite-face effect) for SR than OR faces, demonstrating that SR faces are processed more holistically than OR faces. This differential holistic processing for faces of different races, probably a by-product of visual experience, may be a critical factor in the other-race effect.

Intraneuronal accumulation of hyperphosphorylated protein tau in paired helical filaments together with amyloid-beta peptide (Abeta) deposits confirm the clinical diagnosis of Alzheimer disease. A common cellular mechanism leading to the production of these potent toxins remains elusive. Here we show that, in cultured neurons, membrane depolarization induced a calcium-mediated transient phosphorylation of both microtubule-associated protein tau and amyloid precursor protein (APP), followed by a dephosphorylation of these proteins. Phosphorylation was mediated by glycogen synthase kinase 3 and cyclin-dependent kinase 5 protein kinases, while calcineurin was responsible for dephosphorylation. Following the transient phosphorylation of APP, intraneuronal Abeta accumulated and induced neurotoxicity. Phosphorylation of APP on Thr-668 was indispensable for intraneuronal accumulation of Abeta. Our data demonstrate that an increase in cytosolic calcium concentration induces modifications of neuronal metabolism of APP and tau, similar to those found in Alzheimer disease.

The human amyloid precursor protein (APP) is processed by the nonamyloidogenic and the amyloidogenic catabolic pathways. The sequential cleavage of APP by the beta- and gamma-secretase activities, known as the amyloidogenic processing of APP, leads to the formation of the amyloid-beta peptide (Abeta). Abeta is the main constituent of the amyloid core of senile plaques, a typical hallmark of Alzheimer's disease. In addition to secretases, other cellular proteolytic activities, like the proteasome, might participate in the metabolism of APP. We investigated the consequence of proteasome inhibition on the amyloidogenic processing of human APP. CHO cells and primary cultures of rat cortical neurons expressing human APP or a protein corresponding to its beta-cleaved C-terminal fragment (C99) were treated with lactacystin, an irreversible inhibitor of the chymotrypsin-like activity of the proteasome. Lactacystin significantly decreased the level of Abeta produced from APP in both cellular models, whereas the production of Abeta from C99 was not affected. Lactacystin did not inhibit gamma-secretase activity but was found to inhibit the beta-cleavage of APP, leading to a proportional decrease in Abeta production. Although lactacystin did not inhibit the catalytic activity of recombinant BACE1, a decrease in neuronal beta-secretase activity was measured after treatment with lactacystin.

Small-object manipulation is essential in numerous human activities, although its neural bases are still essentially unknown. Recent functional imaging studies have shown that precision grasping activates a large bilateral frontoparietal network, including ventral (PMv) and dorsal (PMd) premotor areas. To dissociate the role of PMv and PMd in the control of hand and finger movements, we produced, by means of transcranial magnetic stimulation (TMS), transient virtual lesions of these two areas in both hemispheres, in healthy subjects performing a grip-lift task with their right, dominant hand. We found that a virtual lesion of PMv specifically impaired the grasping component of these movements: a lesion of either the left or right PMv altered the correct positioning of fingers on the object, a prerequisite for an efficient grasping, whereas lesioning the left, contralateral PMv disturbed the sequential recruitment of intrinsic hand muscles, all other movement parameters being unaffected by PMv lesions. Conversely, we found that a virtual lesion of the left PMd impaired the proper coupling between the grasping and lifting phases, as evidenced by the TMS-induced delay in the recruitment of proximal muscles responsible for the lifting phase; lesioning the right PMd failed to affect dominant hand movements. Finally, an analysis of the time course of these effects allowed us to demonstrate the sequential involvement of PMv and PMd in movement preparation. These results provide the first compelling evidence for a neuronal dissociation between the different phases of precision grasping in human premotor cortex.

BACKGROUND: Inflammation plays a critical role in lung disease development and progression in cystic fibrosis. Azithromycin is used for the treatment of cystic fibrosis lung disease, although its mechanisms of action are poorly understood. We tested the hypothesis that azithromycin modulates lung inflammation in cystic fibrosis mice. METHODS: We monitored cellular and molecular inflammatory markers in lungs of cystic fibrosis mutant mice homozygous for the DeltaF508 mutation and their littermate controls, either in baseline conditions or after induction of acute inflammation by intratracheal instillation of lipopolysaccharide from Pseudomonas aeruginosa, which would be independent of interactions of bacteria with epithelial cells. The effect of azithromycin pretreatment (10 mg/kg/day) given by oral administration for 4 weeks was evaluated. RESULTS: In naive cystic fibrosis mice, a spontaneous lung inflammation was observed, characterized by macrophage and neutrophil infiltration, and increased intra-luminal content of the pro-inflammatory cytokine macrophage inflammatory protein-2. After induced inflammation, cystic fibrosis mice combined exaggerated cellular infiltration and lower anti-inflammatory interleukin-10 production. In cystic fibrosis mice, azithromycin attenuated cellular infiltration in both baseline and induced inflammatory condition, and inhibited cytokine (tumor necrosis factor-alpha and macrophage inflammatory protein-2) release in lipopolysaccharide-induced inflammation. CONCLUSION: Our findings further support the concept that inflammatory responses are upregulated in cystic fibrosis. Azithromycin reduces some lung inflammation outcome measures in cystic fibrosis mice. We postulate that some of the benefits of azithromycin treatment in cystic fibrosis patients are due to modulation of lung inflammation.

Glucose-induced insulin secretion requires a rise in beta-cell cytosolic Ca2+ ([Ca2+]c) that triggers exocytosis and a mechanistically unexplained amplification of the action of [Ca2+]c. Insulin granules are kept acidic by luminal pumping of protons with simultaneous Cl- uptake to maintain electroneutrality. Experiments using patched, dialyzed beta-cells prompted the suggestion that acute granule acidification by glucose underlies amplification of insulin secretion. However, others found glucose to increase granular pH in intact islets. In this study, we measured islet granular pH with Lysosensor DND-160, a fluorescent dye that permits ratiometric determination of pH < 6 in acidic compartments. Stimulation of mouse islets with glucose reversibly decreased granular pH by mechanisms that are dependent on metabolism and Cl- ions but independent of changes in [Ca2+]c and protein kinase A or C activity. Granular pH was increased by concanamycin (blocker of the vesicular type H+-ATPase) > methylamine (weak base) > Cl- omission. Concanamycin and methylamine did not alter glucose-induced [Ca2+]c increase in islets but strongly inhibited the two phases of insulin secretion. Omission of Cl- did not affect the first phase but decreased the second phase of both [Ca2+]c and insulin responses. Neither experimental condition affected the [Ca2+]c rise induced by 30 mM KCl, but the insulin responses were inhibited by concanamycin > methylamine and not affected by Cl- omission. The amplification of insulin secretion by glucose was not suppressed. We conclude that an acidic granular pH is important for insulin secretion but that the acute further acidification produced by glucose is not essential for the augmentation of secretion via the amplifying pathway.

How fast does the human visual system discriminate individual faces? To address this question, we used a continuous-stimulation paradigm in which event-related potentials (ERPs) to a face stimulus are recorded with respect to another face stimulus, rather than to a preceding blank-screen baseline epoch. Following the shift between two face stimuli, posterior sites showed an early negative ERP deflection that started at 130 ms and peaked at 160 ms, the latency of the N170, an ERP component associated with discriminating faces from objects. The ERP we recorded was larger in amplitude when the preceding stimulus was perceived as a different individual face rather than the same individual face, although face pairs were of equal physical distance in the two conditions. These findings provide direct evidence that individual face discrimination in humans can take place as early as 130 ms following stimulus onset, during the same time window as face detection.