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OBJECTIVES: The REDUC clinical study Part B investigated Vacc-4x/rhuGM-CSF therapeutic vaccination prior to HIV latency reversal using romidepsin. The main finding was a statistically significant reduction from baseline in viral reservoir measurements. Here we evaluated HIV-specific functional T-cell responses following Vacc-4x/rhuGM-CSF immunotherapy in relation to virological outcomes on the HIV reservoir. METHODS: This study, conducted in Aarhus, Denmark, enrolled participants (n = 20) with CD4>500 cells/mm3 on cART. Six Vacc-4x (1.2 mg) intradermal immunizations using rhuGM-CSF (60 µg) as adjuvant were followed by 3 weekly intravenous infusions of romidepsin (5 mg/m2). Immune responses were determined by IFN-γ ELISpot, T-cell proliferation to p24 15-mer peptides covering the Vacc-4x region, intracellular cytokine staining (ICS) to the entire HIVGag and viral inhibition. RESULTS: The frequency of participants with CD8+ T-cell proliferation assay positivity was 8/16 (50%) at baseline, 11/15 (73%) post-vaccination, 6/14 (43%) during romidepsin, and 9/15 (60%)post-romidepsin. Participants with CD8+ T-cell proliferation assay positivity post-vaccination showed reductions in total HIV DNA post-vaccination (p = 0.006; q = 0.183), post-latency reversal (p = 0.005; q = 0.183), and CA-RNA reductions post-vaccination (p = 0.015; q = 0.254). Participants (40%) were defined as proliferation 'Responders' having ≥2-fold increase in assay positivity post-baseline. Robust ELISpot baseline responses were found in 87.5% participants. No significant changes were observed in the proportion of polyfunctional CD8+ T-cells to HIVGag by ICS. There was a trend towards increased viral inhibition from baseline to post-vaccination (p = 0.08). CONCLUSIONS: In this 'shock and kill' approach supported by therapeutic vaccination, CD8+ T-cell proliferation represents a valuable means to monitor functional immune responses as part of the path towards functional HIV cure.
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Vacunas contra el SIDA/inmunología , Vacunas contra el SIDA/uso terapéutico , Antirretrovirales/uso terapéutico , Linfocitos T CD8-positivos/inmunología , Depsipéptidos/uso terapéutico , Seropositividad para VIH/terapia , VIH-1 , Latencia del Virus/inmunología , Adulto , Citocinas/inmunología , Dinamarca , Quimioterapia Combinada , Femenino , Seropositividad para VIH/tratamiento farmacológico , Humanos , Inmunidad Celular , Inmunoterapia , Masculino , Carga Viral/inmunologíaRESUMEN
The measured low frequency vibrational energies of some quantum dots (QDs) deviate from the predictions of traditional elastic continuum models. Recent experiments have revealed that these deviations can be tuned by changing the ligands that passivate the QD surface. This observation has led to speculation that these deviations are due to a mass-loading effect of the surface ligands. In this article, we address this speculation by formulating a continuum elastic theory that includes the dynamical loading by elastic surface ligands. We demonstrate that this model is capable of accurately reproducing the l = 0 phonon energy across a variety of different QD samples, including cores with different ligand identities and epitaxially grown CdSe/CdS core/shell heterostructures. We highlight that our model performs well even in the small QD regime, where traditional elastic continuum models are especially prone to failure. Furthermore, we show that our model combined with Raman measurements can be used to infer the elastic properties of surface bound ligands, such as sound velocities and elastic moduli, that are otherwise challenging to measure.
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We measure the temperature dependence of breathing-mode acoustic vibrations of semiconductor nanocrystals using low-frequency Raman spectroscopy. In CdSe core-only nanocrystals, the lowest-energy l = 0 mode red-shifts with increasing temperature by â¼5% between 77-300 K. Changes to the interatomic bond distances in the inorganic crystal lattice, with corresponding changes to the bulk modulus and density of the material, contribute to the observed energy shift but do not fully explain its magnitude across all nanocrystal sizes. Invariance of the Raman linewidth over the same temperature range suggests that the acoustic breathing mode is inhomogeneously broadened. The acoustic phonons of CdSe/CdS core-shell composite nanocrystals display similar qualitative behavior. However, for large core-shell nanocrystals, we observe a higher-order Raman peak at approximately twice the energy of the l = 0 mode, which we identify as a higher spherical harmonic-the n = 2, l = 0 eigenmode-rather than a two-phonon scattering event.
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Recent experimental and theoretical results have highlighted the surprisingly dominant role of acoustic phonons in regulating dynamic processes in nanocrystals. While it has been known for many years that acoustic phonon frequencies in nanocrystals depend on their size, strategies for tuning acoustic phonon energy at a given fixed size were not available. Here, we show that acoustic phonon frequencies in colloidal quantum dots (QDs) can be tuned through the choice of the surface ligand. Using low-frequency Raman spectroscopy, we explore the dependence of the l = 0 acoustic phonon resonance in CdSe QDs on ligand size, molecular weight, and chemical functionality. On the basis of these aggregated observations, we conclude that the primary mechanism for this effect is mass loading of the QD surface and that interactions between ligands and with the surrounding environment play a comparatively minor yet non-negligible role.
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More effective treatments for major depression are needed. We studied if the selective 5-HT3 receptor antagonist ondansetron can potentiate the antidepressant potential of the selective serotonin (5-HT) reuptake inhibitor (SSRI) paroxetine using behavioral, neurochemical and electrophysiological methods. Flinders Sensitive Line (FSL) rats, treated with ondansetron, and/or a sub-effective dose of paroxetine, were assessed in the forced swim test. The effects of an acute intravenous administration of each compound alone and in combination were evaluated with respect to 5-HT neuronal firing rate in the dorsal raphe nucleus (DRN). Effects of s.c. administration of the compounds alone and in combination on extracellular levels of 5-HT were assessed in the ventral hippocampus of freely moving rats by microdialysis. The results showed that ondansetron enhanced the antidepressant activity of paroxetine in the forced swim test. It partially prevented the suppressant effect of paroxetine on DRN 5-HT neuronal firing and enhanced the paroxetine-induced increase of hippocampal extracellular 5-HT release. These findings indicate that 5-HT3 receptor blockade potentiates the antidepressant effects of SSRIs. Since both paroxetine and ondansetron are used clinically, it might be possible to validate this augmentation strategy in depressed patients.
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Depresión/tratamiento farmacológico , Ondansetrón/farmacología , Paroxetina/agonistas , Inhibidores Selectivos de la Recaptación de Serotonina/agonistas , Antagonistas del Receptor de Serotonina 5-HT3/farmacología , Animales , Depresión/fisiopatología , Modelos Animales de Enfermedad , Núcleo Dorsal del Rafe/efectos de los fármacos , Núcleo Dorsal del Rafe/fisiología , Sinergismo Farmacológico , Fenómenos Electrofisiológicos/efectos de los fármacos , Hipocampo/química , Hipocampo/efectos de los fármacos , Hipocampo/fisiología , Masculino , Microdiálisis , Ratas , Ratas Sprague-Dawley , Serotonina/análisis , Serotonina/fisiologíaRESUMEN
Colloidal quantum dots (QDs) are promising materials for use in solar cells, light-emitting diodes, lasers, and photodetectors, but the mechanism and length of exciton transport in QD materials is not well understood. We use time-resolved optical microscopy to spatially visualize exciton transport in CdSe/ZnCdS core/shell QD assemblies. We find that the exciton diffusion length, which exceeds 30 nm in some cases, can be tuned by adjusting the inorganic shell thickness and organic ligand length, offering a powerful strategy for controlling exciton movement. Moreover, we show experimentally and through kinetic Monte Carlo simulations that exciton diffusion in QD solids does not occur by a random-walk process; instead, energetic disorder within the inhomogeneously broadened ensemble causes the exciton diffusivity to decrease over time. These findings reveal new insights into exciton dynamics in disordered systems and demonstrate the flexibility of QD materials for photonic and optoelectronic applications.
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Glutamate is the main excitatory neurotransmitter in the central nervous system (CNS) and is a major player in complex brain functions. Glutamatergic transmission is primarily mediated by ionotropic glutamate receptors, which include NMDA, AMPA and kainate receptors. However, glutamate exerts modulatory actions through a family of metabotropic G-protein-coupled glutamate receptors (mGluRs). Dysfunctions of glutamatergic neurotransmission have been implicated in the etiology of several diseases. Therefore, pharmacological modulation of ionotropic glutamate receptors has been widely investigated as a potential therapeutic strategy for the treatment of several disorders associated with glutamatergic dysfunction. However, blockade of ionotropic glutamate receptors might be accompanied by severe side effects due to their vital role in many important physiological functions. A different strategy aimed at pharmacologically interfering with mGluR function has recently gained interest. Many subtype selective agonists and antagonists have been identified and widely used in preclinical studies as an attempt to elucidate the role of specific mGluRs subtypes in glutamatergic transmission. These studies have allowed linkage between specific subtypes and various physiological functions and more importantly to pathological states. This article reviews the currently available knowledge regarding the therapeutic potential of targeting mGluRs in the treatment of several CNS disorders, including schizophrenia, addiction, major depressive disorder and anxiety, Fragile X Syndrome, Parkinson's disease, Alzheimer's disease and pain.
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A thin-film transistor: An n-type polymer semiconductor, poly(2,3-bis(perfluorohexyl)thieno[3,4-b]pyrazine), was synthesized through a Pd-catalyzed polycondensation employing a perfluorinated multiphase solvent system. This is the first example of an n-type polymer semiconductor with exclusive solubility in fluorinated solvents. The fabrication of organic field effect transistors containing this new n-type polymer semiconductor is shown.
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1-[2-(2,4-Dimethylphenyl-sulfanyl)-phenyl]-piperazine (Lu AA21004) is a human (h) serotonin (5-HT)(3A) receptor antagonist (K(i) = 3.7 nM), h5-HT(7) receptor antagonist (K(i) = 19 nM), h5-HT(1B) receptor partial agonist (K(i) = 33 nM), h5-HT(1A) receptor agonist (K(i) = 15 nM), and a human 5-HT transporter (SERT) inhibitor (K(i) = 1.6 nM) (J Med Chem 54:3206-3221, 2011). Here, we confirm that Lu AA21004 is a partial h5-HT(1B) receptor agonist [EC(50) = 460 nM, intrinsic activity = 22%] using a whole-cell cAMP-based assay and demonstrate that Lu AA21004 is a rat (r) 5-HT(7) receptor antagonist (K(i) = 200 nM and IC(50) = 2080 nM). In vivo, Lu AA21004 occupies the r5-HT(1B) receptor and rSERT (ED(50) = 3.2 and 0.4 mg/kg, respectively) after subcutaneous administration and is a 5-HT(3) receptor antagonist in the Bezold-Jarisch reflex assay (ED(50) = 0.11 mg/kg s.c.). In rat microdialysis experiments, Lu AA21004 (2.5-10.0 mg/kg s.c.) increased extracellular 5-HT, dopamine, and noradrenaline in the medial prefrontal cortex and ventral hippocampus. Lu AA21004 (5 mg/kg per day for 3 days; minipump subcutaneously), corresponding to 41% rSERT occupancy, significantly increased extracellular 5-HT in the ventral hippocampus. Furthermore, the 5-HT(3) receptor antagonist, ondansetron, potentiated the increase in extracellular levels of 5-HT induced by citalopram. Lu AA21004 has antidepressant- and anxiolytic-like effects in the rat forced swim (Flinders Sensitive Line) and social interaction and conditioned fear tests (minimal effective doses: 7.8, 2.0, and 3.9 mg/kg). In conclusion, Lu AA21004 mediates its pharmacological effects via two pharmacological modalities: SERT inhibition and 5-HT receptor modulation. In vivo, this results in enhanced release of several neurotransmitters and antidepressant- and anxiolytic-like profiles at doses for which targets in addition to the SERT are occupied. The multimodal activity profile of Lu AA21004 is distinct from that of current antidepressants.
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Ansiolíticos/uso terapéutico , Trastorno Depresivo Mayor/tratamiento farmacológico , Piperazinas/uso terapéutico , Sulfuros/uso terapéutico , Animales , Monoaminas Biogénicas/análisis , Citalopram/farmacología , Humanos , Masculino , Ondansetrón/farmacología , Piperazinas/farmacocinética , Piperazinas/farmacología , Ratas , Ratas Sprague-Dawley , Receptor de Serotonina 5-HT1B/metabolismo , Receptores de Serotonina/metabolismo , Reflejo/efectos de los fármacos , Proteínas de Transporte de Serotonina en la Membrana Plasmática/metabolismo , Sulfuros/farmacocinética , Sulfuros/farmacología , Vocalización Animal/efectos de los fármacos , VortioxetinaRESUMEN
AIM: The aim of this study was to estimate healthcare cost and productivity losses as a result of diabetes and diabetes-related chronic complications in Sweden in 1987 and 2005. RESEARCH DESIGN AND METHODS: Published estimates on relative risks and Swedish age-specific diabetes-prevalence rates were used to calculate the proportions of diabetes-related chronic complications that are attributable to diabetes. These attributable risks were applied to cost estimates for diabetes-related chronic complications based on data from Swedish population registers. RESULTS: The estimated total costs for Sweden in 1987 and 2005 were EUR439m and EUR920m, respectively. The increase of 110% was as a result of a 69% increase in the estimated prevalence from 150 000 (1.8% of the population) to 254 000 (2.8%) and of an increase in the estimated annual cost per person diagnosed with diabetes by 24%. Healthcare accounted for 45% of the estimated cost in 1987 and for 37% in 2005. The estimated diabetes-related healthcare cost accounted for approximately 1.0% of total healthcare cost in Sweden in 1987 and for 1.4% in 2005. Diabetes per se accounted for 57% of the healthcare cost in 1987 and for 50% in 2005. The most important chronic complication was cardiovascular disease. CONCLUSIONS: The cost of diabetes is substantial and increasing even in a fairly low-prevalence country such as Sweden. Measures to curb the increase in prevalence and to improve individual control of his or her diabetes seem to be the most important challenges.
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Complicaciones de la Diabetes/economía , Diabetes Mellitus Tipo 1/economía , Diabetes Mellitus Tipo 2/economía , Ausencia por Enfermedad/economía , Costo de Enfermedad , Complicaciones de la Diabetes/epidemiología , Diabetes Mellitus Tipo 1/epidemiología , Diabetes Mellitus Tipo 2/epidemiología , Femenino , Costos de la Atención en Salud/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Sistema de Registros/estadística & datos numéricos , Factores de Riesgo , Ausencia por Enfermedad/estadística & datos numéricos , Suecia/epidemiologíaRESUMEN
We used the validated chronic mild stress (CMS) paradigm to induce anhedonia, a core symptom of major depression, in rats. Thirty percent of animals exposed to CMS are resistant to the development of anhedonia, whereas the remaining are responsive, CMS resilient and CMS sensitive, respectively. We used in situ hybridization to elucidate the molecular mechanisms, which may be involved in the development of anhedonia during CMS. In the CA3 of the ventral hippocampus, we found upregulation of brain-derived neurotrophic factor (BDNF) mRNA in the CMS resilient group indicating protective role of BDNF in stress. Moreover, in the CA3 we found downregulation of vascular endothelial growth factor (VEGF) mRNA in the CMS sensitive group. Downregulation of VEGF suggests impaired hippocampal function, caused by loss of trophic factor neuroprotective support, as part of a previously uncharacterized mechanism for development of anhedonia. CMS induced anhedonia was not related to mRNA expression differences of the dopamine receptors D(1) and D(2), enkephalin, dynorphin, the NMDA receptor subtype NR2B in the ventral striatum, BDNF expression in the dentate gyrus, nor corticotrophin releasing hormone (CRH) and arginine vasopressin (AVP) in the paraventricular nucleus of the hypothalamus. In particular, HPA axis seems to be activated in the CMS resilient group suggesting other pathways protecting against stress sensitivity. We applied the restraint stress procedure to compare effects of a faster and simpler form of stress to CMS and found the latter to be more valid as rats probably easier adapt to restraint stress. Finally, we used the conditioned place preference model to demonstrate a clear tendency towards a distinct morphine induced behavioral difference between CMS resilient and CMS sensitive animals.
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Encéfalo/metabolismo , Depresión/etiología , Depresión/fisiopatología , Hibridación in Situ , Estrés Psicológico/complicaciones , Análisis de Varianza , Animales , Conducta Animal , Encéfalo/patología , Factor Neurotrófico Derivado del Encéfalo/genética , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Condicionamiento Operante/fisiología , Hormona Liberadora de Corticotropina/genética , Hormona Liberadora de Corticotropina/metabolismo , Depresión/patología , Modelos Animales de Enfermedad , Factores de Crecimiento Endotelial/genética , Factores de Crecimiento Endotelial/metabolismo , Masculino , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Ratas , Ratas Wistar , Factores de TiempoRESUMEN
Several lines of research have implicated glutathione (GSH) in schizophrenia. For instance, GSH deficiency has been reported in the prefrontal cortex of schizophrenics in vivo. Further, in rats postnatal GSH-deficiency combined with hyperdopaminergia led to cognitive impairments in the adult. In the present report we studied the effects of 2-day GSH-deficiency with L-buthionine-(S,R)-sulfoximine on monoaminergic function in mice. The effect of GSH-deficiency per se and when combined with the amphetamine and phencyclidine (PCP) models of schizophrenia was investigated. GSH-deficiency significantly altered tissue levels of dopamine (DA), 5-hydroxytryptamine (5-HT) and their respective metabolites homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA) in a region-specific fashion. The effects of GSH-deficiency on tissue monoamines were distinct from and, generally, did not interact with the effects of amphetamine (5 mg/kg; i.p.) on tissue monoamines. Microdialysis studies showed that extracellular DA-release after amphetamine (5 mg/kg, i.p.) was two-fold increased in the nucleus accumbens of GSH-deficient mice as compared with control mice. Basal DA was unaltered. Further, extracellular levels of HVA in the frontal cortex and hippocampus and 5-HIAA in the nucleus accumbens were elevated by GSH-deficiency per se. Spontaneous locomotor activity in the open field was unchanged in GSH-deficient mice. In contrast, GSH-deficiency modulated the locomotor responses to mid-range doses of amphetamine (1.5 and 5 mg/kg, i.p.). Further, GSH-deficient mice displayed an increased locomotor response to low (2 and 3 mg/kg, i.p.) doses of phencyclidine (PCP). In conclusion, the data presented here show that even short-term GSH-deficiency has consequences for DA and 5-HT function. This was confirmed on both neurochemical and behavioral levels. How GSH and the monoamines interact needs further scrutiny. Moreover, the open field findings suggest reduced or altered N-methyl-d-aspartate (NMDA) receptor function in GSH-deficient mice. Thus, GSH-deficiency can lead to disturbances in DA, 5-HT and NMDA function, a finding that may have relevance for schizophrenia.
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Dopamina/metabolismo , Glutatión/deficiencia , Esquizofrenia/fisiopatología , Serotonina/metabolismo , Anfetamina/toxicidad , Animales , Encéfalo/metabolismo , Cromatografía Líquida de Alta Presión , Dopamina/análisis , Alucinógenos/toxicidad , Ácido Homovanílico/metabolismo , Ácido Hidroxiindolacético/metabolismo , Masculino , Ratones , Microdiálisis , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Fenciclidina/toxicidad , Receptores de N-Metil-D-Aspartato/metabolismo , Esquizofrenia/inducido químicamente , Serotonina/análisisRESUMEN
The selective serotonin (5-HT) reuptake inhibitor, citalopram, is a racemic mixture of an S(+)- and R(-)-enantiomer, escitalopram and R-citalopram, respectively. The present study compares the effects of escitalopram, R-citalopram and citalopram on extracellular levels of 5-HT in the frontal cortex of freely moving rats. In addition, co-injection of escitalopram and R-citalopram (ratios 1:2 and 1:4) were assessed. In some experiments escitalopram and R-citalopram were infused into the frontal cortex by reverse microdialysis. Finally, the extracellular level of escitalopram in the frontal cortex was studied after administration of escitalopram alone or in combination with R-citalopram. Escitalopram (1.0-3.9 mg/kg, s.c.) produced a greater maximal increase in extracellular 5-HT than citalopram (2.0-8.0 mg/kg, s.c.). R-citalopram (15.6 mg/kg s.c.) did not affect the 5-HT levels. When co-injected, R-citalopram counteracted the escitalopram-induced increase in extracellular 5-HT levels. Local infusion of the two enantiomers into the frontal cortex produced a similar inhibitory response. R-citalopram did not influence the extracellular levels of escitalopram and therefore does not exert its effect via a pharmacokinetic interaction with escitalopram. In conclusion, the 5-HT-reuptake inhibitory activity of citalopram resides in escitalopram, and the R-enantiomer counteracts this effect. This observation would predict an improved clinical profile of escitalopram compared to citalopram.
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Citalopram/análogos & derivados , Citalopram/farmacología , Espacio Extracelular/efectos de los fármacos , Lóbulo Frontal/efectos de los fármacos , Serotonina/metabolismo , Animales , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Espacio Extracelular/metabolismo , Lóbulo Frontal/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , EstereoisomerismoRESUMEN
Asthmatics are known to react to inhaled hyperosmolar solution. Therefore, the effect of hyperosmolar salt solutions on tight junctions of the airway epithelium was investigated by electron microscopy. Rat trachea was perfused with different concentrations of sodium chloride (NaCl) and then fixed from the luminal side with glutaraldehyde to which the electron dense tracer lanthanum chloride had been added. Lanthanum penetrated 3+/-1% of the tight junctions in trachea perfused with 295 mOsm Krebs-Ringer's buffer (KRB). Adding NaCl to the KRB (KRB-NaCl) increased osmolarity of the solution. After perfusion with 589 or 876 mOsm KRB-NaCl, lanthanum was observed in the lateral intercellular spaces in 50+/-11 and 57+/-6%, respectively. The effect of hyperosmolarity was reversible and only 6+/-1% of the tight junctions were penetrated after perfusion with 295 mOsm KRB solution following 589 mOsm KRB-NaCl perfusion. Adding mannitol to the KRB to an osmolarity of 589 mOsm only caused 5+/-1% of the tight junctions to open, even though osmotic effects were observed. Opening the tight junctions with hyperosmolar salt solutions may play a role in exercise-induced asthma. It may also open the prospect for increased penetration of inhaled drugs into the interstitium and the circulation.
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Solución Salina Hipertónica/farmacología , Uniones Estrechas/efectos de los fármacos , Tráquea/ultraestructura , Animales , Lantano/farmacología , Microscopía Electrónica , Permeabilidad , Ratas , Ratas Sprague-DawleyRESUMEN
The Ca(2+) signaling system in an established immortalized rat parotid acinar cell line, Par-C5, was examined using the Ca(2+)-sensitive fluorescent indicator fura-2 and by measuring inositol 1,4,5-trisphosphate (IP(3)) formation. Agonist-induced increase in intracellular Ca(2+) ([Ca(2+)](i)) by mobilization of intracellular stores and influx across the cell membrane was stimulated by acetylcholine (ACh) and ATP, whereas noradrenaline-(NA)-induced a small [Ca(2+)](i) increase mediated primarily by release from intracellular Ca(2+) stores. [Ca(2+)](i) increase by ACh and ATP was mediated through the phosphoinositide signal pathway since both agonists significantly increased 1,4,5-IP(3) formation and Ca(2+) mobilization was abolished by the phospholipase C inhibitor U73122. In Ca(2+)-free medium, ACh or ATP discharged the IP(3)-sensitive Ca(2+) store and essentially abolished subsequent [Ca(2+)](i) response to thapsigargin (TG). Exposure to ionomycin and monensin after TG induced a further mobilization of Ca(2+), suggesting IP(3)-insensitive stores are present. Furthermore, depletion of IP(3)-sensitive Ca(2+) stores by TG, ACh and ATP enhanced plasmalemmal Ca(2+)-entry pathways. Exposure to tumor necrosis factor-alpha (TNF-alpha), a cytokine associated with lymphocyte invasion of salivary epithelial cells in autoimmune disorders, significantly reduced ACh-stimulated Ca(2+) mobilization. TNF-alpha inhibitory effect on Ca(2+) mobilization was not directly due to an interaction on muscarinic receptors since ACh-induced 1,4,5-IP(3) formation was not altered. These results in the Par-C5 cell line indicate 1) [Ca(2+)](i) is regulated by muscarinic and P2Y-nucleotide receptors and partly by alpha(1)-adrenergic receptors; 2) IP(3)-sensitive and -insensitive Ca(2+) stores exist; 3) Ca(2+) influx activated by ACh, ATP or TG is mediated by the store-operated Ca(2+) entry pathway; and 4) muscarinic agonist-stimulated Ca(2+) mobilization is altered by the cytokine TNF-alpha.
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Señalización del Calcio/fisiología , Calcio/metabolismo , Glándula Parótida/metabolismo , Agonistas alfa-Adrenérgicos/farmacología , Análisis de Varianza , Animales , Calcio/agonistas , Agonistas de los Canales de Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Línea Celular Transformada , Inositol 1,4,5-Trifosfato/biosíntesis , Agonistas Muscarínicos/farmacología , Glándula Parótida/citología , Agonistas del Receptor Purinérgico P2 , Ratas , Receptores Adrenérgicos alfa/fisiología , Receptores Muscarínicos/fisiología , Receptores Purinérgicos P2/fisiología , Receptores Purinérgicos P2Y2 , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Consistent findings in depressed patients are hyperactivity in the hypothalamic-pituitary-adrenal (HPA) axis with high plasma concentrations of adrenocorticotropic hormone and cortisol. Long-term antidepressant treatments seem to normalize this hyperactivity, suggesting a link between the HPA axis and the action of antidepressant treatments. The present study was carried out to study the effects of antidepressant treatments on pro-opiomelanocortin (POMC) mRNA expression, with a focus on interaction with acute stress and 5-HT(1A) receptor activation. Male rats were treated for 21 days with saline, citalopram, fluoxetine, moclobemide or desipramine, and the expression of POMC mRNA in the anterior pituitary was analysed by semi-quantitative in situ hybridization. All antidepressants, but not saline, cocaine and haloperidol, reduced POMC mRNA expression. The decrease in POMC mRNA was not observed until 9 days of citalopram treatment. Decreased POMC mRNA levels were also observed after 14 days of repeated electroconvulsive stimulation. The decreased POMC mRNA levels did not affect the stress-induced POMC mRNA increase, measured following swim stress and restraint stress. Finally, using Fos as a marker for neural activity, we showed attenuation of 8-OH-DPAT-stimulated activity in the paraventricular nucleus following 21 days of citalopram treatment. In conclusion, antidepressant treatments decrease basal POMC mRNA expression without affecting the acute stress response, and the reduced POMC mRNA may be related to reduced 5-HT(1A)-stimulated hypothalamic output.
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Antidepresivos/farmacología , Hipófisis/metabolismo , Proopiomelanocortina/biosíntesis , ARN Mensajero/biosíntesis , Receptores de Serotonina/efectos de los fármacos , Agonistas de Receptores de Serotonina/farmacología , Estrés Psicológico/metabolismo , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Enfermedad Aguda , Animales , Citalopram/farmacología , Genes fos/genética , Inmunohistoquímica , Hibridación in Situ , Masculino , Núcleo Hipotalámico Paraventricular/efectos de los fármacos , Núcleo Hipotalámico Paraventricular/metabolismo , Hipófisis/efectos de los fármacos , Ratas , Ratas Wistar , Receptores de Serotonina 5-HT1 , Restricción Física , Convulsiones/fisiopatología , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Natación/psicologíaRESUMEN
Establishment of salivary cell lines retaining normal morphological and physiological characteristics is important in the investigation of salivary cell function. A submandibular gland cell line, SMG-C6, has recently been established. In the present study, we characterized the phosphoinositide (PI)-Ca2+ signaling system in this cell line. Inositol 1,4,5-trisphosphate(1,4,5-IP3) formation, as well as Ca2+ storage, release, and influx in response to muscarinic, alpha1-adrenergic, P2Y-nucleotide, and cytokine receptor agonists were determined. Ca2+ release from intracellular stores was strongly stimulated by acetylcholine (ACh) and ATP, but not by norepinephrine (NA), epidermal growth factor (EGF), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNFalpha). Consistently, 1, 4,5-IP3 formation was dramatically stimulated by ACh and ATP. ACh-stimulated cytosolic free Ca2+ concentration [Ca2+]i increase was inhibited by ryanodine, suggesting that the Ca2+-induced Ca2+ release mechanism is involved in the ACh-elicited Ca2+ release process. Furthermore, ACh and ATP partially discharged the IP3-sensitive Ca2+ store, and a subsequent exposure to thapsigargin (TG) induced further [Ca2+]i increase. However, exposure to TG depleted the store and a subsequent stimulation with ACh or ATP did not induce further [Ca2+]i increase, suggesting that ACh and ATP discharge the same storage site sensitive to TG. As in freshly isolated submandibular acinar cells, exposure to ionomycin and monensin following ACh or TG induced further [Ca2+]i increase, suggesting that IP3-insensitive stores exist in SMG-C6 cells. Ca2+ influx was activated by ACh, ATP, or TG, and was significantly inhibited by La3+, suggesting the involvement of store-operated Ca2+ entry (SOCE) pathway. These results indicate that in SMG-C6 cells: (i) Ca2+ release is triggered by muscarinic and P2Y-nucleotide receptor agonists through formation of IP3; (ii) both the IP3-sensitive and -insensitive Ca2+ stores are present; and (iii) Ca2+ influx is mediated by the store-operated Ca2+ entry pathway. We conclude that Ca2+ regulation in SMG-C6 cells is similar to that in freshly isolated SMG acinar cells; therefore, this cell line represents an excellent SMG cell model in terms of intracellular Ca2+ signaling.
Asunto(s)
Señalización del Calcio , Glándula Submandibular/metabolismo , Acetilcolina/farmacología , Adenosina Trifosfato/farmacología , Animales , Señalización del Calcio/efectos de los fármacos , Línea Celular , Inositol 1,4,5-Trifosfato/biosíntesis , Modelos Biológicos , Norepinefrina/farmacología , Fosfatidilinositoles/metabolismo , Ratas , Receptores Muscarínicos/efectos de los fármacos , Receptores Muscarínicos/metabolismo , Receptores Purinérgicos P2/efectos de los fármacos , Receptores Purinérgicos P2/metabolismo , Glándula Submandibular/citología , Glándula Submandibular/efectos de los fármacos , Tapsigargina/farmacologíaRESUMEN
A substantial number of patients do not respond sufficiently to antidepressant drugs and are therefore often co-medicated with lithium as an augmentative strategy. Since lithium has been demonstrated to affect 5-HT neurotransmission, we examined the effect of acute and subchronic lithium on 5-HT levels after a challenge with citalopram. We found that subchronic administration of lithium increases extracellular 5-HT baseline level and decreases the extracellular 5-HIAA baseline. However, we found no evidence that the effect of acute citalopram on extracellular 5-HT levels is augmented by acute or subchronic lithium.
Asunto(s)
Citalopram/farmacología , Depresión/tratamiento farmacológico , Espacio Extracelular/efectos de los fármacos , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Litio/farmacocinética , Serotonina/metabolismo , Animales , Esquema de Medicación , Quimioterapia Combinada , Espacio Extracelular/metabolismo , Hipocampo/citología , Litio/sangre , Masculino , Microdiálisis , Ratas , Ratas Sprague-DawleyRESUMEN
Rats were chronically treated with the selective serotonin re-uptake inhibitor citalopram [1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)-5-phtalancarbonitril ], by means of osmotic minipumps. Using an infusion concentration of 50 mg/ml citalopram, steady-state plasma concentrations of approximately 0.3 mcM citalopram were maintained for 15 days. Citalopram plasma levels dropped below pharmacologically active concentrations 48 h after removal of the minipumps. Although chronic treatment with citalopram did induce an attenuated response by extracellular levels of 5-hydroxytryptamine (5-HT) after systemic administration of the 5-HT(1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), no effect of chronic citalopram treatment was observed when 5-HT(1B) receptor function was evaluated with a local infusion of 5-HT(1B/D) receptor agonist, sumatriptan (3-[2-dimethylamino]ethyl-N-methyl-1H-indole-5methane sulphonamide). Controversially, no augmentation of the increase of 5-HT levels was observed upon systemic administration of citalopram. It is concluded that, although chronic treatment with citalopram does induce desensitisation of 5-HT(1A) receptors, the absence of augmented effects of citalopram on 5-HT levels indicates that other mechanisms compensate for the loss of autoreceptor control.
Asunto(s)
Autorreceptores/efectos de los fármacos , Citalopram/farmacocinética , Receptores de Serotonina/efectos de los fármacos , Inhibidores Selectivos de la Recaptación de Serotonina/farmacocinética , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Animales , Citalopram/sangre , Bombas de Infusión , Masculino , Ratas , Ratas Wistar , Receptores de Serotonina 5-HT1 , Serotonina/metabolismo , Agonistas de Receptores de Serotonina/farmacología , Sumatriptán/farmacología , Factores de TiempoRESUMEN
Neuropeptide Y (NPY) has been considered to be involved in the pathogenesis of affective disorders, and chronic treatment with lithium or electroconvulsive stimuli (ECS) has been shown to increase mRNA and peptide levels of NPY in rat brain tissue. Consequently, parameters reflective of NPYergic neurotransmission were studied in the hippocampus of rats following chronic treatment with lithium, ECS or the selective serotonin re-uptake inhibitor (SSRI), citalopram. Lithium (28 days, diet) and ECS (10 days, once daily) treatments caused a marked increase in levels of preproNPY mRNA in the CA1 area and dentate gyrus (DG). This increase was accompanied by an increase in extracellular levels of NPY in the dorsal hippocampus of freely moving rats as determined by microdialysis, suggesting that lithium and ECS treatments lead to an increased biosynthesis and release of NPY in this area. (125)I-peptide YY (PYY) binding was reduced by 40 and 60% respectively in the DG following the same treatments, showing that the increased release is accompanied by a down-regulation of corresponding binding sites. In contrast, citalopram (10 mg/kg i.p., twice daily for 28 days) caused a 100% increase in (125)I-PYY binding in CA, CA3 and DG while levels of preproNPY mRNA and extracellular NPY in the hippocampus were unaffected. The results indicate that various agents and stimuli exerting antidepressant effects in humans, such as chronic lithium, ECS and citalopram all increase NPYergic neurotransmission in the hippocampus by distinct modes of action. Moreover, NPY (6 microg) given intracerebroventricularly (i.c.v.) induced an antidepressant-like effect in the forced swim test. It is hypothesised that the increase in NPYergic neurotransmission may be associated with the mechanism of action of various antidepressant treatments in the alleviation of depression.