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2.
Eng. sanit. ambient ; 26(5): 935-943, set.-out. 2021. tab, graf
Article in Portuguese | LILACS-Express | LILACS | ID: biblio-1346002

ABSTRACT

RESUMO A concepção dos sistemas de drenagem de águas pluviais como separador absoluto é uma característica marcante na gestão das águas urbanas, no entanto não é o reflexo da realidade de tal sistema na maioria das cidades brasileiras, onde frequentemente esgotos e águas pluviais escoam pelos mesmos condutos. Diante disso, o objetivo do presente trabalho foi investigar as contribuições de contaminação fecal dos canais de drenagem afluentes ao Canal do Prado, pertencentes ao sistema de macrodrenagem de águas pluviais da cidade de Campina Grande, na Paraíba. Para a realização da pesquisa de campo, foram definidos sete pontos de monitoramento ao longo do canal e, como indicadores de contaminação fecal, coliformes termotolerantes, Escherichia coli e os ovos dos helmintos Ascaris lumbricoides, Taenia sp, Hymenolepis nana, Hymenolepis diminuta, Enterobius vermicularis, Ancilostomideo sp e Trichuris sp. Os resultados apontaram que ovos de Ascaris lumbricoides foram os que apresentaram maior frequência nos pontos de monitoramento, relacionados à maior descarga de esgotos sanitários. Tais descargas também foram verificadas por elevados valores médios de coliformes termotolerantes que atingiram até 1,6 x 107 UFC.100 mL-1, valor típico de esgoto sanitário. Escherichia coli ocorreu em mais de 90% das amostras coletadas em todos os pontos de monitoramento.


ABSTRACT The design of rainwater drainage systems as an absolute separator is a significant feature in urban water management, however, this is not a reflection of the reality of such a system in most Brazilian cities, where sewage and rainwater often run along the same path. Therefore, the aim of this study was to investigate contributions of fecal contamination from tributary drainage channels to Canal do Prado, belonging to the urban macro-drainage system of rainwater in the city of Campina Grande, Paraíba State. For the fieldwork, seven monitoring points were set along the channel object of this study, determining thermotolerant coliforms, Escherichia coli, and helminth eggs of Ascaris lumbricoides, Taenia sp, Hymenolepis nana, Hymenolepis diminuta, Enterobius vermicularis, and Trichuris sp, as fecal contamination indicators. The results pointed out that eggs of Ascaris lumbricoides were those with the highest frequency in monitoring points, being this related to the increased discharge of sewage. Such discharges were also verified by high mean values of thermotolerant coliforms that reached up to 1.6 x 107 CFU.100 mL-1, typical value of domestic sewage. Escherichia coli was present in over 90% of all samples collected from monitoring points.

3.
J Cell Biol ; 216(10): 3323-3338, 2017 10 02.
Article in English | MEDLINE | ID: mdl-28855251

ABSTRACT

Clathrin-mediated endocytosis (CME) is used to internalize a diverse range of cargo proteins from the cell surface, often in response to specific signals. In neurons, the rapid endocytosis of GluA2-containing AMPA receptors (AMPARs) in response to NMDA receptor (NMDAR) stimulation causes a reduction in synaptic strength and is the central mechanism for long-term depression, which underlies certain forms of learning. The mechanisms that link NMDAR activation to CME of AMPARs remain elusive. PICK1 is a BAR domain protein required for NMDAR-dependent reductions in surface GluA2; however, the molecular mechanisms involved are unclear. In this study, we show that PICK1 makes direct, NMDAR-dependent interactions with the core endocytic proteins AP2 and dynamin. PICK1-AP2 interactions are required for clustering AMPARs at endocytic zones in dendrites in response to NMDAR stimulation and for consequent AMPAR internalization. We further show that PICK1 stimulates dynamin polymerization. We propose that PICK1 is a cargo-specific endocytic accessory protein required for efficient, activity-dependent AMPAR endocytosis.


Subject(s)
Adaptor Protein Complex 2/metabolism , Carrier Proteins/metabolism , Dynamins/metabolism , Endocytosis/physiology , Nuclear Proteins/metabolism , Receptors, AMPA/metabolism , Receptors, N-Methyl-D-Aspartate/metabolism , Adaptor Protein Complex 2/genetics , Animals , Carrier Proteins/genetics , Cytoskeletal Proteins , Dynamins/genetics , HEK293 Cells , Humans , Nuclear Proteins/genetics , Rats , Rats, Wistar , Receptors, AMPA/genetics , Receptors, N-Methyl-D-Aspartate/genetics
4.
Mol Neurobiol ; 53(6): 3513-3527, 2016 08.
Article in English | MEDLINE | ID: mdl-26093381

ABSTRACT

GABA (γ-aminobutyric acid) is the major inhibitory neurotransmitter in the central nervous system, and changes in GABAergic neurotransmission modulate the activity of neuronal networks. Gephyrin is a scaffold protein responsible for the traffic and synaptic anchoring of GABAA receptors (GABAAR); therefore, changes in gephyrin expression and oligomerization may affect the activity of GABAergic synapses. In this work, we investigated the changes in gephyrin protein levels during brain ischemia and in excitotoxic conditions, which may affect synaptic clustering of GABAAR. We found that gephyrin is cleaved by calpains following excitotoxic stimulation of hippocampal neurons with glutamate, as well as after intrahippocampal injection of kainate, giving rise to a stable cleavage product. Gephyrin cleavage was also observed in cultured hippocampal neurons subjected to transient oxygen-glucose deprivation (OGD), an in vitro model of brain ischemia, and after transient middle cerebral artery occlusion (MCAO) in mice, a model of focal brain ischemia. Furthermore, a truncated form of gephyrin decreased the synaptic clustering of the protein, reduced the synaptic pool of GABAAR containing γ2 subunits and upregulated OGD-induced cell death in hippocampal cultures. Our results show that excitotoxicity and brain ischemia downregulate full-length gephyrin with a concomitant generation of truncated products, which affect synaptic clustering of GABAAR and cell death.


Subject(s)
Brain Ischemia/metabolism , Brain Ischemia/pathology , Carrier Proteins/metabolism , Membrane Proteins/metabolism , Neurons/metabolism , Neurons/pathology , Receptors, GABA-A/metabolism , Animals , Calpain/metabolism , Cell Death , Cells, Cultured , Down-Regulation/drug effects , Glucose/deficiency , Glutamic Acid/toxicity , Male , Mice, Inbred C57BL , Neurotoxins/toxicity , Oxygen , Rats, Wistar , Synapses/drug effects , Synapses/metabolism
5.
EMBO Rep ; 15(5): 548-56, 2014 May.
Article in English | MEDLINE | ID: mdl-24723684

ABSTRACT

MicroRNAs fine-tune gene expression by inhibiting the translation of mRNA targets. Argonaute (Ago) proteins are critical mediators of microRNA-induced post-transcriptional silencing and have been shown to associate with endosomal compartments, but the molecular mechanisms that underlie this process are unclear, especially in neurons. Here, we report a novel interaction between Ago2 and the BAR-domain protein, PICK1. We show that PICK1 promotes Ago2 localization at endosomal compartments in neuronal dendrites and inhibits Ago2 function in translational repression following neuronal stimulation. We propose that PICK1 provides a link between activity-dependent endosomal trafficking and local regulation of translation in neurons.


Subject(s)
Argonaute Proteins/metabolism , Carrier Proteins/metabolism , Dendrites/metabolism , Endosomes/metabolism , Nuclear Proteins/metabolism , Animals , COS Cells , Carrier Proteins/genetics , Cells, Cultured , Chlorocebus aethiops , Dendrites/genetics , Gene Expression Regulation , HEK293 Cells , Humans , Lim Kinases/biosynthesis , Lim Kinases/genetics , Mice , MicroRNAs/genetics , Nuclear Proteins/genetics , Protein Biosynthesis/genetics , RNA Interference , RNA, Small Interfering , Receptors, N-Methyl-D-Aspartate/biosynthesis , Receptors, N-Methyl-D-Aspartate/metabolism
6.
Cell Mol Life Sci ; 69(16): 2691-715, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22382927

ABSTRACT

In addition to its central roles in protein quality control, regulation of cell cycle, intracellular signaling, DNA damage response and transcription regulation, the ubiquitin-proteasome system (UPS) plays specific roles in the nervous system, where it contributes to precise connectivity through development, and later assures functionality by regulating a wide spectrum of neuron-specific cellular processes. Aberrations in this system have been implicated in the etiology of neurodevelopmental and neurodegenerative diseases. In this review, we provide an updated view on the UPS and highlight recent findings concerning its role in normal and diseased nervous systems. We discuss the advantages of the model organism Caenorhabditis elegans as a tool to unravel the major unsolved questions concerning this biochemical pathway and its involvement in nervous system function and dysfunction, and expose the new possibilities, using state-of-the-art techniques, to assess UPS function using this model system.


Subject(s)
Caenorhabditis elegans/metabolism , Nervous System Physiological Phenomena , Neurodegenerative Diseases/metabolism , Proteasome Endopeptidase Complex/metabolism , Ubiquitin/metabolism , Animals , Caenorhabditis elegans/genetics
7.
PLoS One ; 5(4): e10139, 2010 Apr 12.
Article in English | MEDLINE | ID: mdl-20405034

ABSTRACT

Glutamic acid decarboxylase is responsible for synthesizing GABA, the major inhibitory neurotransmitter, and exists in two isoforms--GAD65 and GAD67. The enzyme is cleaved under excitotoxic conditions, but the mechanisms involved and the functional consequences are not fully elucidated. We found that excitotoxic stimulation of cultured hippocampal neurons with glutamate leads to a time-dependent cleavage of GAD65 and GAD67 in the N-terminal region of the proteins, and decrease the corresponding mRNAs. The cleavage of GAD67 was sensitive to the proteasome inhibitors MG132, YU102 and lactacystin, and was also abrogated by the E1 ubiquitin ligase inhibitor UBEI-41. In contrast, MG132 and UBEI-41 were the only inhibitors tested that showed an effect on GAD65 cleavage. Excitotoxic stimulation with glutamate also increased the amount of GAD captured in experiments where ubiquitinated proteins and their binding partners were isolated. However, no evidences were found for direct GADs ubiquitination in cultured hippocampal neurons, and recombinant GAD65 was not cleaved by purified 20S or 26S proteasome preparations. Since calpains, a group of calcium activated proteases, play a key role in GAD65/67 cleavage under excitotoxic conditions the results suggest that GADs are cleaved after ubiquitination and degradation of an unknown binding partner by the proteasome. The characteristic punctate distribution of GAD65 along neurites of differentiated cultured hippocampal neurons was significantly reduced after excitotoxic injury, and the total GAD activity measured in extracts from the cerebellum or cerebral cortex at 24h postmortem (when there is a partial cleavage of GADs) was also decreased. The results show a role of the UPS in the cleavage of GAD65/67 and point out the deregulation of GADs under excitotoxic conditions, which is likely to affect GABAergic neurotransmission. This is the first time that the UPS has been implicated in the events triggered during excitotoxicity and the first molecular target of the UPS affected in this cell death process.


Subject(s)
Glutamate Decarboxylase/metabolism , Hippocampus/cytology , Neurons/enzymology , Proteasome Endopeptidase Complex/metabolism , Animals , Cells, Cultured , Glutamic Acid/pharmacology , Hydrolysis/drug effects , Rats , Ubiquitination
8.
Open educational resource in Portuguese | CVSP - Brazil | ID: cfc-181870

ABSTRACT

Falou sobre a capacitação de profissionais do Serviço Social da Indústria da Construção do Rio de Janeiro, que se tornaram multiplicadores em seus locais de trabalho, e também sobre as Brigadas Antidengue montadas em 56 obras do município. "Os operários receberam instruções teóricas e práticas, o que nos ajudou a reduzir significativamente o número de focos no município. Os resíduos de construção civil representam hoje mais de 20% dos locais propícios à procriação da larva do mosquito. Em primeiro lugar, com 31,5%, estão as caixas d'água mal vedadas". Os arquivos estão disponíveis para leitura, audição e/ou download nos ícones ao lado.

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