Fluoxetine Treatment in Rats Increases the Rate of Taurine Transport in Mononuclear Cells.

Fluoxetine, a selective serotonin reuptake inhibitor antidepressant, modulates the mitogen-induced proliferation of lymphocytes. Lymphocytes contain taurine and express taurine transporter (TauT). Among the effects of taurine on lymphocytes are protection against oxidants and regulation of the inflammatory aspects of the immune response. Our aim was to determine the influence of fluoxetine treatment on taurine transport, and to determine the presence of TauT in the mononuclear cells of rats. METHODS: Male adult Sprague-Dawley rats were treated with fluoxetine 10 mg/kg i.p. for 1, 2, and 3 weeks. The cells were obtained by density gradients. [3H]Taurine was used for transport assays. Amino acid levels were determined by high-performance liquid chromatography. Immunolabeling of CD4+, CD8+, and TauT was performed. The mRNA of TauT was evaluated by RT-PCR. Controls were included for each protocol. RESULTS: The transport of taurine, after 1 week of treatment, was significantly augmented compared to controls. The affinity significantly increased at 1 and 2 weeks. While the percentage of CD4+ cells decreased and that of CD8+ cells increased, the percentage of TauT in CD4+ and CD8+ cells was not affected. Reduction of levels of aspartic acid, glutamic acid, threonine, alanine, glycine, and arginine occurred at 1 and 2 weeks. The taurine concentration significantly decreased after 2 and 3 weeks of treatment. The estimation of mRNA of TauT was not different. CONCLUSION: Taurine transport increases with fluoxetine treatment, and so this could be related to an immunomodulatory role of fluoxetine through TauT. Inhibition of serotonin reuptake might be involved in the regulation of taurine transport in mononuclear cells.

Serotonin transporter in lymphocytes of rats exposed to physical restraint stress.

OBJECTIVES: Glucocorticoids and stress cause transcriptional and functional changes on the serotonin transporter (SERT) in the central nervous system. Stress can produce specific modifications of SERT in lymphocytes, which could be associated with alterations in immune response. The aim of this study was to evaluate the effect of a physical restraint stress protocol on (1) rat lymphocyte proliferation in the presence of the selective serotonin reuptake inhibitor fluoxetine and (2) SERT kinetic parameters, i.e. binding capacity (Bmax), affinity (Kd) and Hill coefficient (nH). METHODS: Male adult Sprague-Dawley rats were placed in Plexiglass boxes (5 h daily for 5 days), and blood was obtained by cardiac puncture on day 6. Serum corticosterone was quantitated by an immunoenzymatic assay. Lymphocytes were isolated by density gradients and adhesion to plastic, of which there was sufficient material for further experiments, then cultured with or without the mitogen concanavalin A (Con A, 2 µg/ml) and fluoxetine (1-50 µM). Cell proliferation was measured with tetrazolium salts, and [(3)H]paroxetine was used as a SERT-specific ligand for binding assays. RESULTS: Restraint produced a significant increase in serum corticosterone of stressed rats. The proliferative response to Con A was similar in the controls and stressed animals. Fluoxetine reduced cell proliferation with and without Con A. Restraint diminished the inhibitory effect of fluoxetine on proliferation. Restraint also increased Bmax and Kd, but decreased nH. Treatment of rats with actinomycin D, a transcription inhibitor, reduced Bmax in stressed animals. CONCLUSIONS: Restraint stress modulated the effect of fluoxetine on cell proliferation, probably through the modification of the presence and the function of SERT.

Efecto de la restricción física: sobre el papel de los receptores 5-HT1A en la proliferación linfocitaria / Effect of phisical restraints: the role of the 5-HT1A in lymphocyte proliferation

El estrés afecta el sistema inmunológico, sin embargo, no hay reportes sobre receptores de serotonina en linfocitos. Se estudiaron los receptores 5-HT1A y la proliferación linfocitaria de ratas macho adultas luego de restricción física. Fueron colocadas en cajas de Plexiglass durante 5 horas diarias por 1, 3 ó 5 días. Se extrajo sangre cardíaca, se aislaron los linfocitos por gradiente de densidades y adhesión al plástico; se cultivaron con el agonista 8-hidroxi-di-n-propil-aminotetralina(8-OH-DPAT) ó el antagonista N-(2-(4-(2-metoxifenil)-1-piperazinetil)-N-(2-piridinil) ciclohexanocarboxamida (WAY-100635) de los receptores 5-HT1A y del mitógeno concanavalina A. La proliferación se midió con sales de tetrazolio. La 8-OH-DPAT no afectó la proliferación. El WAY-100635 disminuyó la proliferación. La restricción física aumentó la sensibilidad al efecto del WAY-100635, lo cual podría deberse a cambios en la expresión o a una modulación funcional de los receptores por efecto del estrés, como se ha reportado previamente en cerebro de rata.

Stress affects the immune system, however, little is known about the effects on specific modifications of lymphocytes serotonin receptors. The effects of restraint stress on the role of 5-HT1A receptors in lymphocyte proliferation were evaluated in male adult rats. They were placed in Plexiglass boxes, during 5 daily hours for 1, 3 or 5 consecutive days. Next day a blood sample was obtained by cardiac punction. Lymphocytes were isolated by density gradient and adhesionto plastic. They were cultured with agonist 8-hydroxy-di-n-propyl-aminotetralin (8-OH-DPAT) or antagonist N-(2-(4-(2-methoxyphenyl)-1-piperazinyl) ethyl)-N-(2-pyridiyl) cyclohexanecarboxamide (WAY-100635) of 5-HT1A receptors and the mitogen concanavalin A. Proliferation was measured by tetrazolium salts. 8-OH-DPAT did not modify cell proliferation; WAY-100635 diminished it. Restraint stress increased the susceptibility to effect of WAY-100635. These results suggest changes in the expression or functional modulation of 5-HT1A receptors in lymphocytes by stress, similar to previous reports on serotonergic system in rat brain.

Ácidos grasos omega-3 como coadyuvantes del tratamiento antidepresivo y sus efectos sobre el factor neurotrófico derivado del cerebro en suero y en células mononucleares / Omega-3 fatty acids as adjunctive of antidpressant therapy and its effects on brain-derived neurotrophic factor in serum, monocytes and lymphocytes

El estrés disminuye el factor neurotrófico derivado del cerebro (BDNF) en el hipocampo; los antidepresivos y los ácidos grasos omega-3 podrían aumentarlo. En pacientes con depresión mayor, investigamos la respuesta clínica a un antidepresivo solo o con ácido eicosapentanoico (EPA), y su influencia sobre los niveles de BDNF. Veinte pacientes se diagnósticaron según el DSM-IV-TR; evaluamos la respuesta con la Escala de Hamilton para Depresión (HAM-D). Los controles fueron 15 sujetos sanos. Los pacientes se distribuyeron en 2 grupos: uno recibió fluoxetina 20 mg/día y EPA 3.000 mg/día, y otro con fuoxetina 20 mg/día y placebo, durante 8 semanas. Se tomaron muestras de sangre en las semanas 0 y 8 para obtener suero, sin anticuoagulante, para medir los niveles de BDNF, y para aislamiento de células mononucleares, con heparina, para la localización inmunocitoquímica de BDNF. Diez pacientes no continuaron por diferentes causas. De los 10 restantes, 5 recibieron EPA y 5, placebo. El BDNF sérico disminuyó despúes de tratamiento en ambos grupos, más evidente en el grupo con EPA en el análisis pareado. El porcentaje de células mononucleares que expresaron BDNF fue inferior en los pacientes y aumentó después de los tratamientos. Por otra parte, las células con BDNF, las cuales estaban bajas en los deprimidos, aumentaron después de los tratamientos, lo que indica que en estás, el papel del tratamiento, especialmente con la combinación del antidepresivo y el ácido en cuestión, las modificaciones en el BDNF son más evidentes. A pesar de la limitante que constituye la inclusión de los sujetos diagnósticados, más la permanencia de los incluidos, los resultados significativos señalan un efecto beneficiosos del uso de EPA en la depresión mayor

Stress is associated with a decreased expression of brainderived neurotrophic factor (BDNF) in the hippocampus. Antidepressants and omega-3 fatty acids might increase circulating BDNF. This research was done to evaluate, in major depression patients, the possible differences in clinical response to an antidepressant alone or in combination with eicosapentaenoic acid (EPA), and their influence on BDNF levels. Nineteen patients were diagnosed according to DSM-IV-TR criteria; severity and response was evaluated by Hamilton Depression Rating Scale (HAM-D). Control group was composed of 15 healthy subjects. Patients were randomized on a double-blind bassis in two groups: one recived fluoxetine 20 mg/day and EPA 3,000 mg/day, and the other one fluoxetine 20 mg/day an placebo, during 8 weeks. Blood samples were taken for obtaining serum and for isolating monocytes and lymphocytes at weeks 0 and 8. Ten patients dropped out for different causes. Of the remaining 9 subjetcs, 4 recived EPA and 5 got placebo. The percentage of mononuclear cells expressing BDNF was lower in patients, and it was significantly increased after the treatments. EPA seems to augment the clinical response. In depressed, after treatment, there was a lower content of serum BDNF. Moreover, mononuclear cells with BDNF, which were lower in this group of depressed, increased after the treatments, indicating that in cells the modulation of BDNF by antidepressants is more evident

Taurine and proliferation of lymphocytes in physically restrained rats.

BACKGROUND: Taurine is present in lymphocytes and seems to modulate certain immune cell functions. Among the effects of taurine on these cells are protection against antioxidants and regulation of inflammatory aspects of the immune response. Stress affects antigen presentation, traffic and proliferation of leukocytes, as well as antibody and cytokine secretion. The purposes of this study were to explore the possible direct effects of taurine concentrations on lymphoproliferation and interleukins levels in control and in physical restrained rats. METHODS: Lymphocytes of male Sprague-Dawley rats, stressed by physical restrain and controls (5 h per day for 5 days) were isolated from blood by Histopaque (1077 g/l) and differential adhesion to plastic, and then cultured (72 h) in the presence of different concentrations of taurine (0.5-50 mM), beta-alanine (0.5-50 mM), or both, without or with the T cells mitogen, concanavalin A. Plasma and lymphocytes levels of pro-inflammatory interleukin-1beta and anti-inflammatory interleukin-10 were respectively measured by Pierce Endogen rat ELISA Kits. Taurine in plasma and in lymphocytes were determined by HPLC. RESULTS: Lymphoproliferation of resting cells significantly decreased in the presence of 3 and 6 mM taurine and increased up to control level at 12 mM taurine. In concanavalin A-activated lymphocytes, the effect of taurine was greater. beta-alanine increased lymphoproliferation in a bell shaped dose-dependent manner and decreased it in activated lymphocytes but in a lower magnitude. In combination, beta-alanine impaired the effect of taurine at 3 and 6 mM. After restriction, no change in lymphoproliferation was observed at different concentrations of the amino acids without or with concanavalin A, although pro-inflammatory interleukin and taurine in plasma and in lymphocytes significantly increased. CONCLUSIONS: Taurine affects lymphoproliferation in control rats, following a dose-dependent manner, an effect that might involve its transport into the cells. Elevation of interleukin-1beta produced in stressed rats by physical restrain could seriously affect the immune balance, whereas taurine increase might be protective. These results suggest that taurine and taurine transport play a role in lymphoproliferation. In addition, modifications of taurine system in lymphocytes take place during restriction stress.

Fluoxetine treatment to rats modifies serotonin transporter and cAMP in lymphocytes, CD4+ and CD8+ subpopulations and interleukins 2 and 4.

Several lines of evidences indicate that antidepressants produce various immunomodulatory effects. Fluoxetine, an antidepressant and selective serotonin reuptake inhibitor, modulates immune cells in vitro. To explore the in vivo influence of fluoxetine on lymphocytes, male Sprague-Dawley rats were treated daily, 10 mg/kg, or with saline solution for 1, 2 and 3 weeks. The presence of serotonin transporter in CD3+, CD4+ and CD8+ subpopulations of T lymphocytes was determined by immunofluorescence. Serotonin transporter was also labeled with [(3)H]paroxetine, specific binding defined with imipramine. Plasma levels of pro-inflammatory interleukin 2 (IL-2), and anti-inflammatory interleukin 4 (IL-4), were measured by ELISA; and cAMP concentration by radioimmunoassay. Fluoxetine significantly increased the number of lymphocytes expressing serotonin transporter and elevated the binding of [(3)H]paroxetine. The percentage of CD4+ cells decreased, that of CD8+ increased, and CD3+ did not change. The ratio CD4+/CD8+ was significantly lowered. Fluoxetine administration elevated the levels of IL-4 at 1, 2 and 3 weeks; and of IL-2, at 2 and 3 weeks. IL-4/IL-2 ratio was significantly increased in fluoxetine group respecting the controls and was similar during the 3 weeks of treatment. Fluoxetine produced a significant decrease in cAMP concentrations in lymphocytes, probably by secondary activation of serotonin receptors. Treatment with fluoxetine modified immune parameters in plasma and lymphocytes of rats, which might be relevant for its systemic therapeutic action as an antidepressant.

Taurine transporter in lymphocytes of patients with major depression treated with venlafaxine plus psychotherapy.

The taurine transporter and taurine are present in lymphocytes, where taurine functions as an antioxidant and an anti-inflammatory agent. Taurine levels are elevated in lymphocytes of subjects with major depression, but returns to control levels after treatment with the antidepressant mirtazapine. Patients (40) were diagnosed using the Diagnostic and Statistical Manual IV of the American Psychiatric Association, and the severity of their condition was determined by the Hamilton Scale of Depression. One group of patients was treated with venlafaxine and the other with venlafaxine plus Neuro-Linguistic Programming. Lymphocytes were isolated from the peripheral blood by Ficoll/Hypaque. The coexistence of the taurine transporter with a subpopulation of CD4+ and CD8+ lymphocytes was measured by immunofluorescence. The levels of the pro-inflammatory, IL-2, and the anti-inflammatory, IL-4, cytokines were determined by ELISA while plasma amino acid levels were determined by HPLC. The percentage of CD4+ cells significantly decreased after both treatments, whereas the levels of CD8+ cells remained unchanged. The taurine transporter of CD4+ and CD8+ cells decreased after integrate treatment. No differences were found in the levels of IL-2 while IL-4 levels increased after integrate treatment. The observed effects of treatment on the taurine transporter and IL-4 content might modify lymphocyte activity during depression.

Serotonin transporter is differentially localized in subpopulations of lymphocytes of major depression patients. Effect of fluoxetine on proliferation.

Modifications of lymphocyte serotonergic system have been described in major depression. The aim of this study was to determine new possible changes of this system in depression. Twenty eight patients, free of drugs, diagnosed with major depression disorder by Structured Clinical Interview for Disorders of Axis I, without medical illnesses, written consent, approved by Ethical Committees were included. Controls were 30 healthy subjects without family history of psychiatric disease. Blood monocytes were isolated with Ficoll/Hypaque, and lymphocytes by differential adhesion to plastic. Serotonin and 5-hydroxyindoleacetic acid determined by HPLC. Monocytes had higher serotonin concentrations than lymphocytes, and serotonin/5-hydroxyindoleacetic acid was lower in patients. Basal proliferation was elevated in depressed and not increased by Concanavalin A. Fluoxetine reduced basal proliferation more efficiently in patients, indicating activation of lymphocytes in depression. The number of cells expressing serotonin transporter was reduced in depressed. There were no differences in CD4+ (approximately 50%) or CD8+ (approximately 25%) lymphocytes between the groups, although CD8+ were lower in depressed, and greater number of them co-localized serotonin transporter than CD4+, which could be crucial for function in relation to serotonin and its receptors in immune cells. Lymphocytes were activated in this group of patients and fluoxetine reduced proliferation, probably being relevant for the psychopharmacological treatment of depression.

Serotonin, 5-HT1A serotonin receptors and proliferation of lymphocytes in major depression patients.

Serotonin receptors are present in lymphocytes and might be related to the functionality of these cells in health and in pathology. The serotonergic system is affected in the brain and in peripheral immune cells of depressed patients. The objectives of this work were to evaluate the basal proliferation of lymphocytes, the response to the mitogen concanavalin A, and the role of serotonin 5-HT(1A) receptors. Twenty-nine patients, 19-52 years old, were diagnosed for a major depression episode with the Statistical and Diagnostic Manual-IV of the American Psychiatric Association, approved by ethic committees and gave written consent. The Hamilton depression score was 30.60 +/- 2.65. An apparently healthy group without a family history of psychiatric illness was included. Blood peripheral lymphocytes were isolated by density gradients with Ficoll/Hypaque and differential adhesion to plastic, cultured in 96-well plaques with RPMI-1640 medium with or without 4 mug/ml of concanavalin A. 8-Hydroxy-2-(di-n-propylamino)tetralin (5-40 nM) and WAY-100,478 (0.1-100 microM), agonist and antagonist of 5-HT(1A) receptors, serotonin (12.5-100 nM) or imipramine (0.1-100 microM) were also added. Proliferation was evaluated at 72 h with 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyltetrazolium bromide, and the optical density was 570 nm. Basal proliferation was three times higher in depressed patients than in controls, whereas no response to mitogen was obtained, and 5-HT(1A) receptors significantly reacted to the agonist, with increases of about 31-54% at 10, 20 and 40 nM of the specific agonist, indicating initial activation probably in relation to autoimmunity and overreactivity of these receptors in depression. The antagonist reduced proliferation in mitogen-stimulated lymphocytes, 50% in controls and 70% in depressed patients, with a differential concentration dependency; probably, these receptors are more sensitive in depression due to increased 5-HT(1A) receptor transduction. The antagonist also reduced the stimulation produced by the 5-HT(1A) agonist. Imipramine caused biphasic effects according to concentrations, showing a possible dual role for serotonin, although all values were significantly higher in depressed subjects. The described alterations might be of relevance in the pathophysiology of depression.

Taurine transport and transporter localization in peripheral blood lymphocytes of controls and major depression patients.

Taurine concentration is increased in peripheral blood lymphocytes in a group of depressed subjects. After this observation [3H]taurine transport was measured in lymphocytes of 10 major depression patients (19-60 years old), diagnosed by the criteria of the American Psychiatric Association, with moderate severity as determined by Hamilton Scale for Depression (32 +/- 6). The control group comprised 10 subjects (20-56 years old). Taurine transporter and CD4+ (helpers) and CD8+ (cytolytic) T lymphocytes were immunolabeled. No significant differences were observed in immunochemical analyses. CD4+ cells were 53% and CD8+ cells 28% of the total lymphocytes in both controls and patients. Two taurine transport components, high and low affinity, were demonstrated in both groups. Taurine transporter was present in 16% of CD4+ and CD8+ lymphocytes in controls and patients. This preliminary report exhibits the presence of taurine transporter in peripheral blood lymphocytes but no differences were observed between controls and depressed subjects.

Acido fólico y vitamina B12 en niños, adolesdentes y mujeres embarazadas en Venezuela / Folic acid and vitamin B12 in children, adolescents and pregnant women in Venezuela

Las consecuencias de la deficiencia de ácido fólico y vitamina B12 en el apropiado desarrollo y funcionamiento del organismo están bien documentadas. Este trabajo tiene como objetivo determinar la magnitud de la deficiencia de ácido fólico y vitamina B12 en grupos vulnerables, pertenecientes a los estratos socioeconómicos bajos (Graffar IV y V) en Venezuela. La muestra proviene de tres encuestas de Condiciones de Vida, cuyo trabajo de campo ejecutó Fundacredesa, entre los años 2001 y 2003 en 14 ciudades del país, en la Gran Caracas y en el Estado Vargas. Se procesaron 5658 muestras de suero y se determinó ácido fólico y vitamina B12 por radio inmuno ensayo, de infantes, niños, adolescentes y mujeres embarazadas de los estratos más pobres. La prevalencia de deficiencia de ácido fólico varió entre 27,5 a 81,79 por ciento en los diferentes grupos estudiados. La deficiencia de vitamina B12 en la muestra de las principales ciudades (Nacional) fue de 11,4 por ciento, pero existió también un porcentaje similar de exceso de vitamina B12 sérica. La prevalencia de deficiencia de ácido fólico y vitamina B12 en embarazadas de la Gran Caracas fue de 36,32 y 61,34 por ciento, respectivamente. La prevalencia de deficiencia de ácido fólico fue alta, especialmente en mujeres en edad reproductiva, adolescentes embarazadas y en la población estudiada en el Estado Vargas. Es perentorio que esta situación reciba una intervención inmediata, por medio de programas de suplementación a los grupos vulnerables y la ampliación del espectro de la fortificación de alimentos con este nutriente

Taurine effect on neuritic outgrowth from goldfish retinal explants in the absence and presence of fetal calf serum.

Post-crush goldfish retinal explants were cultured in the absence of fetal calf serum and in the presence of calcium, albumin, glucose or taurine. There was an increase in length and density of the neurites in a dose-dependent manner when fetal calf serum was added to the medium. The addition of calcium to the basic medium, Leibovitz, produced a small increase in neuritic outgrowth at low concentrations, but inhibited outgrowth at higher levels. Albumin did not significantly modify neuritic outgrowth. Glucose, at variable concentrations, produced a bell-shaped increase in length and density of the neurites. Taurine, in the absence of fetal calf serum, produced a small increase in neuritic outgrowth at 10 mM. In the presence of glucose, taurine stimulated outgrowth was less efficient than in the presence of fetal calf serum. In order to investigate the mechanisms of action of taurine as a trophic agent, the culture medium in the absence of fetal calf serum will be supplemented with glucose.

Cambios clinicos y hematologicos en becerros infectados con Anaplasma marginale / Clinical and hematological change in calves infected with Anaplasma marginale

Clinical and hematological changes of six Anaplasma marginale(isolated Zulia) inoculated calves (experimental group) and four healthy calves (control group) were studied during twenty and eighty days before and after infection, respectively. The behavior of the four calves used as control group was stable and no significant changes in the parameters analyzed was observed. The experimental group developed the three typical phases of illness. During the prepatent phase, which lasted a mean of 21.2 + /- 2.56 days, the animals were asymptomatic and no significant changes in the hematological values occurred, but a remarkable transitory decrease in number of lymphocytes from 6.5 x 10(6) to 3.3 x 10(6) cells/ml. The infection during the acute phase produced a highly severe effect in two animals, a severe effect in three animals and a mild effect in one. The effects observed were the following: 1) a fast decrease in haematocrite, ranging from 6 to 10 percent; 2) values of parasitaemia varied from 15 to 48 percent 3) a greater body temperature than the control animals (40.5 vs. 38.5 degrees C); 4) a elevated heart frequency, from 60 to 110 beats/min; 5) an increase in the concentration of neotrophiles from 10 x 10(6) to 13 x 10(6) cells/ml; 6) The number of monocytes also augmented from 3 x 10(6) to 6 x 10(6) cells/ml; and 7) an important decrease of weight gain. The natural course of infection was interrupted with oxytetracycline when the haematocrite of the animal lowered to values less or equal to 10 percent. Then, the animals showed a rapid recovery with an undetectable parasitaemia and concomitant return to basal line of the rest of the parameters