RESUMO
Landfills often become a source of environmental impact due to the percolation of the leachate (liquid resulting from the decomposition of buried waste), which can eventually disperse and contaminate the soil and water bodies. This study aimed to evaluate quantitatively and spatially the leachate plume from the former Belo Horizonte landfill in the Coqueiros and Taiobas watersheds by using tritium (3H) concentration in nineteen (19) sampling points: 14 groundwater, 4 surface water and 1 leachate. Among the methodologies applied to determine the 3H content in the leachate, pre-filtration proved to be the most effective due to the ease treating of the sample, as well as the identical results obtained compared to the traditional method. The 3H contents resulted in concentrations up to thirty times higher in the leachate (=96,19 to 111,87 TU) and ten times in groundwater (PM23/3 = 45,06 TU), compared to the calculated threshold (TS) of rainfall tritium (TS_3Hrain = 3,41 TU). Temporal series results indicate no trend and no influence of seasonality regarding the entry of pluvial tritium, leachate liquid level and wells water level. The 3H contour maps shows a transport of the mixture (water and leachate) towards the Coqueiros stream, with a higher concentration of leachate in wells closer to the landfill slope (PM13, PM14, PM15, and PM23) and in the southwest margin of the Coqueiros stream (PM17). In the furthest portion of the landfill, 3H levels are higher than the base threshold only at the deepest level of PM20 (=3,90 TU), possibly due to factors such as: i) denser character of the leachate compared to groundwater; ii) greater influence of recharge for regions further away from the landfill slope; and iii) the presence of the rock about 50 m from the surface. Based on the hydrogeological conceptual model, the transport of the leachate water mixture in the groundwater water compartment of the Coqueiros watershed basin seems to follow a piston or dispersion flow, with a small Peclet number (Pd = 0,05, as an example), as indicated by the high correlation coefficient values (R2 ≈ 0,85 to 0,99) from graphic interpolation. The mixture of water and leachate results in a transient time (tt) ≈ 30 years and linear velocity (Vl) ≈ 3-14 m/year in the interpolated sections. These indicate a coherent correlation with the mean hydraulic conductivity (K ≈ 29,34 m/year), as they result in effective porosity values between (Ne) ≈ 0.16 to 0.36, which corresponds to Ne range for unconsolidated sediments, similar to the wells lithotype profiles. Time series evaluation combined with lumped parameter approach suggest to be an interesting way to better understanding the HCM, and, therefore, to determine hydrodynamic parameters using tritium as a tracer.
Assuntos
Água Subterrânea , Monitoramento de Radiação , Eliminação de Resíduos , Poluentes Químicos da Água , Poluentes Químicos da Água/análise , Trítio/análise , Brasil , Instalações de Eliminação de Resíduos , Água , Monitoramento Ambiental/métodosRESUMO
Intracellular levels of cyclic nucleotide second messengers are regulated predominantly by a large superfamily of phosphodiesterases (PDEs). Most of the different PDE variants play specific physiological functions; in fact, PDEs can associate with other proteins allowing them to be strategically anchored throughout the cell. In this regard, precise cellular expression and compartmentalization of these enzymes produce the specific control of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) gradients in cells and enable their integration with other signaling pathways.In trypanosomatids, some PDEs are essential for their survival and play fundamental roles in the adaptation of these parasites to different environmental stresses, as well as in the differentiation between their different life cycle forms. Given that these enzymes not only are similar to human PDEs but also have differential biochemical properties, and due to the great knowledge of drugs that target human PDEs, trypanosomatid PDEs could be postulated as important therapeutic targets through the repositioning of drugs.In this chapter, we describe a simple and sensitive radioisotope-based method to measure cyclic 3',5'-nucleotide phosphodiesterase using [3H]cAMP.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/isolamento & purificação , Ensaios Enzimáticos/métodos , Marcação por Isótopo/métodos , Proteínas de Protozoários/isolamento & purificação , Trypanosoma cruzi/metabolismo , 3',5'-AMP Cíclico Fosfodiesterases/química , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , AMP Cíclico/química , AMP Cíclico/metabolismo , Estágios do Ciclo de Vida , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismo , Transdução de Sinais , Trítio/químicaRESUMO
Karst environments have an inherent complexity that interferes with their hydrogeology comprehension. Hence, isotope hydrology can be a valuable tool to assess trajectory of subsurface flows in an unexplored setting. The study area is located in the Lagoa Santa Karst, an environmental protection area of great economic, cultural and ecological importance, where Neoproterozoic metalimestones accommodate karst-fractured aquifers, characterized by complex water dynamics, essential vulnerability and high productivity. The purpose of this study was to investigate groundwater flow origins of springs using principally environmental stable isotopes 2H and 18O. Rainwater and spring water were sampled and analysed. The LMWL presents angular and linear coefficients strongly similar to those of the GMWL. Spring isotopic signatures, which represent the base flow and present wide-ranging of 2H and 18O, were separated into two groups. The first group can be associated with recent rainwater major contributions, while the second group shows significant evaporated water contributions, largely represented by resurgences. Tritium concentration and physico-chemical parameter data supported this interpretation, pointing that waters of the second group remained more time on the surface and subsurface. Therefore, using isotope tracers to evaluate upper groundwater zone in this tropical karst system is a powerful instrument for water resources management.
Assuntos
Deutério/análise , Monitoramento Ambiental/métodos , Água Subterrânea/química , Nascentes Naturais/química , Isótopos de Oxigênio/análise , Brasil , Hidrologia , Trítio/análise , Recursos HídricosRESUMO
Purpose: We analyzed the molecular mechanisms leading to glutamate release from rat primary cultures of RPE cells, under isosmotic conditions. Thrombin has been shown to stimulate glutamate release from astrocytes and retinal glia; however, the effect of thrombin on glutamate release from RPE cells has not been examined. Our previous work showed that upon the alteration of the blood-retina barrier, the serine protease thrombin could contribute to the transformation, proliferation, and migration of RPE cells. In this condition, elevated extracellular glutamate causes neuronal loss in many retinal disorders, including glaucoma, ischemia, diabetic retinopathy, and inherited photoreceptor degeneration. Methods: Primary cultures of rat RPE cells were preloaded with 1 µCi/ml 3H-glutamate in Krebs Ringer Bicarbonate (KRB) buffer for 30 min at 37 °C. Cells were rinsed and super-perfused with 1 ml/min KRB for 15 min. Stable release was reached at the 7th minute, and on the 8th minute, fresh KRB containing stimuli was added. Results: This study showed for the first time that thrombin promotes specific, dose-dependent glutamate release from RPE cells, induced by the activation of protease-activated receptor 1 (PAR-1). This effect was found to depend on the Ca2+ increase mediated by the phospholipase C-ß (PLC-ß) and protein kinase C (PKC) pathways, as well as by the reverse activity of the Na+/Ca2+ exchanger. Conclusions: Given the intimate contact of the RPE with the photoreceptor outer segments, diffusion of RPE-released glutamate could contribute to the excitotoxic death of retinal neurons, and the development of thrombin-induced eye pathologies.
Assuntos
Cálcio/metabolismo , Ácido Glutâmico/metabolismo , Proteína Quinase C/metabolismo , Epitélio Pigmentado da Retina/citologia , Trocador de Sódio e Cálcio/metabolismo , Trombina/farmacologia , Fosfolipases Tipo C/metabolismo , Animais , Forma Celular/efeitos dos fármacos , Transportador 1 de Aminoácido Excitatório/metabolismo , Fragmentos de Peptídeos/farmacologia , Transporte Proteico/efeitos dos fármacos , Ratos Long-Evans , Receptor PAR-1/metabolismo , Transdução de Sinais/efeitos dos fármacos , Trítio/metabolismoRESUMO
Serotonin (5-HT) has been recognized as a neurotransmitter in the vertebrate retina, restricted mainly to amacrine and bipolar cells. It is involved with synaptic processing and possibly as a mitogenic factor. We confirm that chick retina amacrine and bipolar cells are, respectively, heavily and faintly immunolabeled for 5-HT. Amacrine serotonergic cells also co-express tyrosine hydroxylase (TH), a marker of dopaminergic cells in the retina. Previous reports demonstrated that serotonin transport can be modulated by neurotransmitter receptor activation. As 5-HT is diffusely released as a neuromodulator and co-localized with other transmitters, we evaluated if 5-HT uptake or release is modulated by several mediators in the avian retina. The role of different glutamate receptors on serotonin transport and release in vitro and in vivo was also studied. We show that L-glutamate induces an inhibitory effect on [3H]5-HT uptake and this effect was specific to kainate receptor activation. Kainate-induced decrease in [3H]5-HT uptake was blocked by CNQX, an AMPA/kainate receptor antagonist, but not by MK-801, a NMDA receptor antagonist. [3H]5-HT uptake was not observed in the presence of AMPA, thus suggesting that the decrease in serotonin uptake is mediated by kainate. 5-HT (10-50 µM) had no intrinsic activity in raising intracellular Ca2+, but addition of 10 µM 5-HT decreased Ca2+ shifts induced by KCl in retinal neurons. Moreover, kainate decreased the number of bipolar and amacrine cells labeled to serotonin in chick retina. In conclusion, our data suggest a highly selective effect of kainate receptors in the regulation of serotonin functions in the retinal cells.
Assuntos
Ácido Caínico/farmacologia , Retina/metabolismo , Serotonina/metabolismo , Animais , Cálcio/metabolismo , Células Cultivadas , Embrião de Galinha , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Neurotransmissores/metabolismo , Receptores de Glutamato/metabolismo , Receptores de Ácido Caínico/metabolismo , Retina/citologia , Retina/efeitos dos fármacos , Retina/embriologia , Neurônios Retinianos/efeitos dos fármacos , Neurônios Retinianos/metabolismo , Trítio/metabolismoRESUMO
Substantia nigra pars reticulata is the output station in basal ganglia; its GABAergic neurons control the activity of thalamo-cortical premotor nuclei, thus controlling motor behavior. D1-like and D2-like presynaptic dopamine receptors on subthalamo-nigral afferents by modulation of glutamate release change the firing rate of nigral neurons; however, their relative contribution to the control of glutamate release and their pharmacological properties have not been studied. This is important since the prevalence of the inhibition or stimulation of release determines the firing rate of nigral neurons, therefore motor activity. Here we used depolarization induced [3H]-glutamate release in slices of rat substantia nigra from reserpinized and non-reserpinized rats to explore the relative contribution of the D1-like and D2-like receptor subtypes to the control of glutamate release. We found a significant control of release by D1-like and D3R, and a modest effect of D2R. D4R exerted no effect. Dopamine showed more potency for D3R than for D1-like receptors; however, these latter enhanced release to a greater degree, as shown by the Emax. We also co-activated these to test their interaction; an antagonist interaction of D1-like with D2 and D3R, and an additive between D2 and D3R were found. Pharmacological receptor antagonist effects in release from reserpinized vs. non-reserpinized slices were similar, suggesting that endogenous dopamine stimulates receptors in the same way. These findings suggest differences in the control of glutamate release by different dopamine receptors in the substantia nigra, which could contribute to explaining the effect of dopamine and its agonists on motor behavior.
Assuntos
Ácido Glutâmico/metabolismo , Terminações Pré-Sinápticas/metabolismo , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D3/metabolismo , Substância Negra/metabolismo , Trítio/metabolismo , Animais , Dopamina/farmacologia , Relação Dose-Resposta a Droga , Masculino , Técnicas de Cultura de Órgãos , Terminações Pré-Sinápticas/efeitos dos fármacos , Ratos , Ratos Wistar , Receptores de Dopamina D1/agonistas , Receptores de Dopamina D3/agonistas , Substância Negra/efeitos dos fármacosRESUMO
Traumatic brain injury (TBI) is a devastating disease frequently followed by behavioral disabilities including post-traumatic epilepsy (PTE). Although reasonable progress in understanding its pathophysiology has been made, treatment of PTE is still limited. Several studies have shown the neuroprotective effect of creatine in different models of brain pathology, but its effects on PTE is not elucidated. Thus, we decided to investigate the impact of delayed and chronic creatine supplementation on susceptibility to epileptic seizures evoked by pentylenetetrazol (PTZ) after TBI. Our experimental data revealed that 4â¯weeks of creatine supplementation (300â¯mg/kg, p.o.) initiated 1â¯week after fluid percussion injury (FPI) notably increased the latency to first myoclonic and tonic-clonic seizures, decreased the time spent in tonic-clonic seizure, seizure intensity, epileptiform discharges and spindle oscillations induced by a sub-convulsant dose of PTZ (35â¯mg/kg, i.p.). Interestingly, this protective effect persists for 1â¯week even when creatine supplementation is discontinued. The anticonvulsant effect of creatine was associated with its ability to reduce cell loss including the number of parvalbumin positive (PARV+) cells in CA3 region of the hippocampus. Furthermore, creatine supplementation also protected against the reduction of GAD67 levels, GAD activity and specific [3H]flunitrazepam binding in the hippocampus. These findings showed that chronic creatine supplementation may play a neuroprotective role on brain excitability by controlling the GABAergic function after TBI, providing a possible new strategy for the treatment of PTE.
Assuntos
Lesões Encefálicas Traumáticas/complicações , Creatina/farmacologia , Epilepsia Pós-Traumática/complicações , Epilepsia Pós-Traumática/prevenção & controle , Neurônios GABAérgicos/efeitos dos fármacos , Convulsões/complicações , Convulsões/prevenção & controle , Animais , Lesões Encefálicas Traumáticas/tratamento farmacológico , Lesões Encefálicas Traumáticas/patologia , Ondas Encefálicas/efeitos dos fármacos , Região CA3 Hipocampal/metabolismo , Região CA3 Hipocampal/patologia , Morte Celular/efeitos dos fármacos , Creatina/uso terapêutico , Epilepsia Pós-Traumática/tratamento farmacológico , Flunitrazepam/metabolismo , Glutamato Descarboxilase/metabolismo , Masculino , Fármacos Neuroprotetores/uso terapêutico , Pentilenotetrazol , Ensaio Radioligante , Ratos , Convulsões/induzido quimicamente , Fatores de Tempo , Trítio/metabolismoRESUMO
The subcommissural organ (SCO) is an ancient and conserved brain gland secreting into cerebrospinal fluid (CSF) glycoproteins that form the Reissner fiber (RF). The present investigation was designed to further investigate the dynamic of the biosynthetic process of RF glycoproteins prior and after their release into the CSF, to identify the RF proteome and N-glycome and to clarify the mechanism of assembly of RF glycoproteins. Various methodological approaches were used: biosynthetic labelling injecting 35S-cysteine and 3H-galactose into the CSF, injection of antibodies against galectin-1 into the cerebrospinal fluid, light and electron microscopical methods; isolated bovine RF was used for proteome analyses by mass spectrometry and glycome analysis by xCGE-LIF. The biosynthetic labelling study further supported that a small pool of SCO-spondin molecules rapidly enter the secretory pathways after its synthesis, while most of the SCO-spondin molecules are stored in the rough endoplasmic reticulum for hours or days before entering the secretory pathway and being released to assemble into RF. The proteomic analysis of RF revealed clusterin and galectin-1 as partners of SCO-spondin; the in vivo use of anti-galectin-1 showed that this lectin is essential for the assembly of RF. Galectin-1 is not secreted by the SCO but evidence was obtained that it would be secreted by multiciliated ependymal cells lying close to the SCO. Further, a surprising variety and complexity of glycan structures were identified in the RF N-glycome that further expands the potential functions of RF to a level not previously envisaged. A model of the macromolecular organization of Reissner fiber is proposed.
Assuntos
Glicoproteínas/metabolismo , Órgão Subcomissural/fisiologia , Animais , Bovinos , Cisteína/metabolismo , Citoplasma/metabolismo , Epêndima/citologia , Epêndima/metabolismo , Galactose/metabolismo , Galectina 1/metabolismo , Glicoproteínas/ultraestrutura , Glicosilação , Masculino , Polissacarídeos/química , Polissacarídeos/metabolismo , Ratos Sprague-Dawley , Via Secretória , Coloração e Rotulagem , Órgão Subcomissural/ultraestrutura , Radioisótopos de Enxofre/metabolismo , Trítio/metabolismoRESUMO
The neonatal exposure to general anesthetics has been associated with neuronal apoptosis and dendritic spines morphologic changes in the developing brain. Ketamine, a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, is widely used in pediatric patients to induce general anesthesia, analgesia, and perioperative sedation. In the present study, we investigated short- and long-term effects of a single ketamine (20 mg/kg, s.c.) neonatal exposure at postnatal day 7 in rats on the hippocampal and frontal cortical cellular viability. Additionally, putative neurochemical alterations and neurobehavioral impairments were evaluated in the adulthood. Ketamine neonatal administration selectively decreased cellular viability in the hippocampus, but not in the frontal cortex, 24 h after the treatment. Interestingly, a single ketamine neonatal exposure prevented the vulnerability to glutamate-induced neurotoxicity in the frontal cortex of adult rats. No short- or long-term damage to cellular membranes, as an indicative of cell death, was observed in hippocampal or cortical slices. However, ketamine induced a long-term increase in hippocampal glutamate uptake. Regarding behavioral analysis, neonatal ketamine exposure did not alter locomotor activity and anxiety-related parameters evaluated in the open-field test. However, ketamine administration disrupted the hippocampal-dependent object recognition ability of adult rats, while improved the motor coordination addressed on the rotarod. These findings indicate that a single neonatal ketamine exposure induces a short-term reduction in the hippocampal, but not in cortical, cellular viability, and long-term alterations in hippocampal glutamate transport, improvement on motor performance, and short-term recognition memory impairment.
Assuntos
Sistema X-AG de Transporte de Aminoácidos/metabolismo , Comportamento Animal/efeitos dos fármacos , Antagonistas de Aminoácidos Excitatórios/toxicidade , Lobo Frontal/metabolismo , Hipocampo/metabolismo , Ketamina/toxicidade , Animais , Animais Recém-Nascidos , Comportamento Exploratório/efeitos dos fármacos , Feminino , Ácido Glutâmico/farmacocinética , Ácido Glutâmico/toxicidade , Técnicas In Vitro , Masculino , Ratos , Ratos Wistar , Reconhecimento Psicológico/efeitos dos fármacos , Natação , Trítio/farmacocinéticaRESUMO
Striatal dopamine D2 receptors activate the PLCâ¯ââ¯IP3â¯ââ¯Calcineurin-signaling pathway to modulate the neural excitability of En+ Medium-sized Spiny GABAergic neurons (MSN) through the regulation of L-type Ca2+ channels. Presynaptic dopaminergic D2 receptors modulate GABA release at striatopallidal terminals through L-type Ca2+ channels as well, but their signaling pathway is still undetermined. Since D2 receptors are Gi/o-coupled and negatively modulate adenylyl cyclase (AC), we investigated whether presynaptic D2 receptors modulate GABA release through the same signaling cascade that controls excitability in the striatum or by the inhibition of AC and decreased PKA activity. Activation of D2 receptors stimulated formation of [3H]IP1 and decreased Forskolin-stimulated [3H]cAMP accumulation in synaptosomes from rat Globus Pallidus. D2 receptor activation with Quinpirole in the presence of L 745,870 decreased, in a dose-dependent manner, K+-induced [3H]GABA release in pallidal slices. The effect was prevented by the pharmacological blockade of Gi/o ßγ subunit effects with Gallein, PLC with U 73122, IP3 receptor activation with 4-APB, Calcineurin with FK506. In addition, when release was stimulated with Forskolin to activate AC, D2 receptors also decreased K+-induced [3H]GABA release, an effect occluded with the effect of the blockade of PKA with H89 or stimulation of release with the cAMP analog 8-Br-cAMP. These data indicate that D2 receptors modulate [3H]GABA release at striatopallidal terminals by activating the PLCâ¯ââ¯IP3â¯ââ¯Calcineurin-signaling cascade, the same one that modulates excitability in soma. Additionally, D2 receptors inhibit release when AC is active. Both mechanisms appear to converge to regulate the activity of presynaptic L-type Ca2+ channels.