The impact of patients with generalized anxiety disorder in the Brazilian Private Healthcare System: a claim database study with expert's perspective / O impacto de pacientes com transtorno de ansiedade generalizada no Sistema Único de Saúde: um estudo de base de sinistros com a perspectiva de especialistas

This study describes the healthcare resource use and costs associated with anxiety assessing claim database outcomes and expert opinion from the perspective of the Brazilian Private Healthcare System. A retrospective analysis of the Orizon database was conducted, containing claims data of anxiety patients reported in Private Healthcare Systems (2015-2017) according to the ICD-10 code (F40 or F41 and their subtypes). Further, a 3-hour online meeting brought together five anxiety and Health Insurance Companies experts to discuss their perspectives. The total cost of the 18,069 patients identified in the database was BRL 490 million: related to medical appointments (2%), exams (16%), emergency room (5%), and others (77%). The mean number of appointments was 5.1 in a 4-year period, performed by 61% of the patients. Approximately 2,595 visits were made to psychiatrists by 923 patients, and 95% underwent at least one examination (100.6 examinations per patient, on average). The identification of anxiety patients and their corresponding burden is challenging to estimate. The higher impact is related to the frequency of healthcare use before the diagnosis than the treatment itself. These outcomes may help plan and implement adequate healthcare programs for patients with anxiety.

Este estudo descreve o uso de recursos de saúde e os custos relacionados à ansiedade associando resultados de uma base de dados administrativa e opinião de especialistas na perspectiva do Sistema Privado de Saúde Brasileiro. Foi realizada uma análise retrospectiva da base de dados da Orizon de pacientes com ansiedade em atendimento hospitalizar ou ambulatorial no Sistema Privado de Saúde (2015 - 2017) com o código CID-10 (F40 ou F41 e os seus subtipos), adicionalmente promovemos uma reunião online de 3 horas com cinco especialistas em ansiedade e em seguros de saúde para discutir as suas perspectivas. O custo total dos 18.069 pacientes identificados no banco de dados foi de R$ 490 milhões, relacionados a consultas médicas (2%), exames (16%), pronto-socorro (5%) e outros (77%). A média de consultas foi de 5,1 em um período de 4 anos, realizadas por 61% dos pacientes. Aproximadamente 2.595 visitas foram feitas a psiquiatras por 923 pacientes, e 95% realizaram pelo menos um exame (média de 100,6 exames por paciente. É desafiador identificar e estimar o impacto da doença no Sistema Privado de Saúde Brasileiro. O impacto maior está relacionado à frequência de uso de serviços de saúde antes do diagnóstico, em comparação com o próprio tratamento. Esses resultados podem ajudar a planejar e implementar programas de saúde adequados para pacientes com ansiedade.

The impact of depression in the Brazilian private healthcare system: patient journey and economic burden study associating claim database and expert's opinion approach / O impacto da depressão no sistema de saúde privado brasileiro: estudo da jornada do paciente e da carga econômica associando banco de dados de sinistros e abordagem de opinião de especialistas

Objective: The study aimed to describe the profile and economic burden of patients with depression from the perspective of the Brazilian Private Healthcare System (PHS). Methods: A two-step methodological quantitative-qualitative research design was performed: retrospective descriptive analysis of the Orizon database of patients with at least one claim of depression (F33, F38, or F39) in PHS (2013-2019) and experienced physicians perspective from an expert meeting. Results: 1,802 patients fulfilling the eligibility criteria counted BRL 74,978 million across the 4-year period. Over this period, nearly 60% of patients had a medical appointment (6.6 appointments per patient, on average), 61% had a psychologist, 9.8% had a psychiatrist appointment, and an average of 115.2 exams and 8.7 emergency visits per patient were performed. According to the experts, the economic impact of depression is more significant when considering the indirect costs related to productivity loss and impairment in occupational and interpersonal functioning. Conclusion: Identifying and diagnosing patients with depression and their real burden is challenging; even with significant costs identified in the claim database analyses in the Brazilian PHS, the real impact must be higher if indirect costs are considered. The depressive disorder should be prioritized in the Brazilian PHS to establish more adequate health policies.

Objetivo: O estudo teve como objetivo descrever o perfil e a carga econômica de pacientes com depressão na perspectiva do Sistema Único de Saúde (SUS). Métodos: Foi realizado um projeto de pesquisa quantitativo-qualitativo metodológico em duas etapas: análise descritiva retrospectiva do banco de dados Orizon de pacientes com pelo menos uma alegação de depressão (F33, F38 ou F39) no PHS (2013- 2019) e perspectiva de médicos experientes de uma reunião de especialistas. Resultados: 1.802 pacientes que preencheram os critérios de elegibilidade totalizaram R$ 74,978 milhões no período de 4 anos. Nesse período, cerca de 60% dos pacientes tiveram consulta médica (6,6 consultas por paciente, em média), 61% tiveram psicólogo, 9,8% consulta com psiquiatra e foram realizados em média 115,2 exames e 8,7 atendimentos de emergência por paciente . Segundo os especialistas, o impacto econômico da depressão é mais significativo quando considerados os custos indiretos relacionados à perda de produtividade e prejuízo no funcionamento ocupacional e interpessoal. Conclusão: Identificar e diagnosticar pacientes com depressão e sua real carga é desafiador; mesmo com custos significativos identificados nas análises da base de sinistros do SUS brasileiro, o impacto real deve ser maior se considerados os custos indiretos. O transtorno depressivo deve ser priorizado na APS brasileira para o estabelecimento de políticas de saúde mais adequadas.

Feedback facilitation by adenosine A2A receptors of ATP release from mouse hippocampal nerve terminals.

The adenosine modulation system is mostly composed by inhibitory A1 receptors (A1R) and the less abundant facilitatory A2A receptors (A2AR), the latter selectively engaged at high frequency stimulation associated with synaptic plasticity processes in the hippocampus. A2AR are activated by adenosine originated from extracellular ATP through ecto-5'-nucleotidase or CD73-mediated catabolism. Using hippocampal synaptosomes, we now investigated how adenosine receptors modulate the synaptic release of ATP. The A2AR agonist CGS21680 (10-100 nM) enhanced the K+-evoked release of ATP, whereas both SCH58261 and the CD73 inhibitor α,ß-methylene ADP (100 µM) decreased ATP release; all these effects were abolished in forebrain A2AR knockout mice. The A1R agonist CPA (10-100 nM) inhibited ATP release, whereas the A1R antagonist DPCPX (100 nM) was devoid of effects. The presence of SCH58261 potentiated CPA-mediated ATP release and uncovered a facilitatory effect of DPCPX. Overall, these findings indicate that ATP release is predominantly controlled by A2AR, which are involved in an apparent feedback loop of A2AR-mediated increased ATP release together with dampening of A1R-mediated inhibition. This study is a tribute to María Teresa Miras-Portugal.

Tratamento cirúrgico por abordagem transoral da laringocele interna com laser de diodo / Surgical treatment by transoral approach of laryngocele internal with diode laser

Laringocele é a dilatação anormal do sáculo laríngeo, que se estende lateralmente às pregas vestibulares e mantém comunicação com o lúmen laríngeo. Não há consenso em relação às modalidades cirúrgicas para o tratamento. A utilização de laser no tratamento das lesões laringe permite ressecções com maior precisão e menor dano tecidual. Objetivo: apresentar série de casos de pacientes com diagnóstico de laringocele interna, submetidos a tratamento cirúrgico por abordagem transoral com laser de diodo. Método: realizado estudo descritivo e retrospectivo no Serviço de Otorrinolaringologia do Hospital do Servidor Público Municipal de São Paulo, no período de abril de 2021 a julho de 2023, de quatro pacientes com diagnóstico de laringocele interna que foram submetidos a tratamento cirúrgico por abordagem transoral com laser de diodo. Resultados: dos quatro pacientes, dois eram do sexo masculino e dois do sexo feminino, com idade que variou de 46 a 71 anos e com média etária de 63,2 anos. Todos foram diagnosticados com laringocele interna unilateral e submetidos a marsupialização da laringocele por abordagem transoral com laser de diodo. No seguimento pós-operatório não houve recorrências das laringoceles. Conclusão: nas laringoceles internas o tratamento cirúrgico pela via transoral tem se mostrado a melhor opção, pela menor morbimortalidade, menor tempo cirúrgico e menor tempo de internação. A abordagem transoral com o laser de diodo é uma alternativa ao laser de CO2, tendo sido observado nesta série de casos uma adequada hemostasia durante o procedimento cirúrgico. Palavras-chave: Laringocele. Laringopiocele. Terapêutica. Terapia a laser.

Increased ATP release and CD73-mediated adenosine A2A receptor activation mediate convulsion-associated neuronal damage and hippocampal dysfunction.

Extracellular ATP is a danger signal to the brain and contributes to neurodegeneration in animal models of Alzheimer's disease through its extracellular catabolism by CD73 to generate adenosine, bolstering the activation of adenosine A2A receptors (A2AR). Convulsive activity leads to increased ATP release, with the resulting morphological alterations being eliminated by A2AR blockade. However, it is not known if upon convulsions there is a CD73-mediated coupling between ATP release and A2AR overactivation, causing neurodegeneration. We now show that kainate-induced convulsions trigger a parallel increase of ATP release and of CD73 and A2AR densities in synapses and astrocytes of the mouse hippocampus. Notably, the genetic deletion of CD73 attenuates neuronal degeneration but has no impact on astrocytic modifications in the hippocampus upon kainate-induced convulsions. Furthermore, kainate-induced convulsions cause a parallel deterioration of hippocampal long-term potentiation (LTP) and hippocampal-dependent memory performance, which is eliminated by knocking out CD73. This demonstrates the key role of the ATP release/CD73/A2AR pathway to selectively control synaptic dysfunction and neurodegeneration following an acute brain insult, paving the way to consider CD73 as a new therapeutic target to prevent neuronal damage upon acute brain damage.

Enumeration, characterisation and clinicopathological significance of circulating tumour cells in patients with colorectal carcinoma.

BACKGROUND: The purposes of the study were to enumerate and characterise the circulating tumour cell (CTC) and cluster/micro-emboli (CTM) in blood from patients with colorectal carcinoma (CRC) as well as to investigate their clinical relevance. METHODS: Peripheral blood of six healthy donors (control) and sixty-two patients with CRC were collected to isolate CTCs by an immunomagnetic negative selection approach. EPCAM and cytokeratin 18 (CK18) antibodies were used to identify the CTCs. The size and the phenotypic variations were evaluated to characterise these isolated CTCs. Additionally, mRNA expressions of the CTCs and the corresponding primary carcinoma were assessed using a multi-gene panel to determine the cellular heterogeneities between CTCs and primary carcinoma. RESULTS: We detected CTCs and CTMs in 72% (41/57) and 32% (18/57) of the patients with CRC, respectively. The total number and length were significantly higher (p<0.0001) in the CTCs than the CTMs. CTCs, especially EPCAMPositiveCK18Posositve subclones, were detected more in the patients with advanced pathological cancer stages when compared to those with early cancer stages (mean: 12.5 vs 4.0, p=0.0068). mRNA profiling of CTCs unveiled three different CTC subtypes expressing epithelial, epithelium-mesenchymal transition (EMT) and stemness signatures, which were different from those of the primary carcinoma. The expressions of EPCAM, HRAS, BRAF, TP53, SLUG, TWIST1, CD44 and MMP9 of CTCs were altered when compared to the primary tumours in patients with CRC. CONCLUSION: Our findings provide insights into the biology of the CTC, presence of heterogeneous CTC populations in CRC and differential expression of genes in different pathological stages of CTC which can improve the management of patients with CRC.

Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma.

The aim of the present study was to isolate and investigate the genetic heterogeneities in single circulating tumour cells (CTCs) from patients with colorectal carcinoma (CRC). Twenty-eight single CTCs were collected from eight patients with CRC using a negative immunomagnetic enrichment method. After validation with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene expression in 3 colon cancer cell lines, a panel of 19 genes were used to analyse the single CTCs (n = 28), primary colorectal carcinoma tissues (n = 8) and colon carcinoma cells (n = 6) using real-time qPCR. Genetic heterogeneities were assessed by comparing gene expression profiles of single CTCs from the different patients and in the same patient, respectively. Genetic profiling of the single CTCs showed extensive heterogeneities of the selected genes among the CTCs. Hierarchical clustering analyses exhibited two clusters of CTCs with differentially expressed genes, which highlighted different modifications from the primary carcinomas. Further, the genetic heterogeneities were observed between different patients or in the same patient. Finally, AKT1 expression was significantly (p = 0.0129) higher in single CTCs from CRC of advanced pathological stages (III or IV) CRC than in CTCs from CRC of early stages (I or II). Our findings suggest that single-cell genetic analysis can monitor the genetic heterogeneities and guide the personalised therapeutic targets in clinical sectors.

Planning and building Health Care Networks in Brazil's Federal District. / O Planejamento e a construção das Redes de Atenção à Saúde no DF, Brasil.

This article reports on the experience of implementing Health Care Planning (HCP) in the territories of Itapoã, Paranoá and São Sebastião in the East Region of Brazil's Federal District. HCP began at the end of 2016 with Itapoã and was expanded to the other territories in 2018. The results point to a better organised health care network, specifically as regards care for chronic conditions, hypertension and diabetes. The activities involved a series of thematic theory workshops and tutoring workshops carried out in Primary Health Care (PHC) and Specialised Ambulatory Care (SAC) facilities. In PHC, macro-processes (territorialisation, family registration, risk stratification, family risk classification, local diagnosis, care by block of hours, elimination of waiting times, and others) were organised to support meeting certain of the population's demands. In SAC, an Ambulatory Specialities Clinic was set up using the technology of continuous care provided by a multi-professional team to high- and very high-risk hypertensive and diabetic patients stratified in PHC, and care provision is shared. One of the strong points in the integration of PHC and SAC was matrix support provided by SAC professionals in "laboratory units". HCP has been an important management tool for organising health care in the East Region.

O artigo objetiva descrever o planejamento e a construção das Redes de Atenção à Saúde (RAS), por meio da Planificação da Atenção à Saúde (PAS), na Região Leste, Distrito Federal. Trata-se de um relato de experiência sobre a PAS, realizada de 2016 a 2018. As atividades foram desenvolvidas a partir de um conjunto de oficinas teóricas temáticas, de tutorias realizadas na Atenção Primária à Saúde (APS) e na Atenção Ambulatorial Especializada (AAE). Os resultados apontam uma melhor organização da RAS, especificamente para a linha de cuidado das condições crônicas, hipertensão e diabetes. Na APS foram organizados os macroprocessos: territorialização, cadastramento das famílias, estratificação de risco, classificação de riscos familiares, diagnóstico local, atendimento por bloco de horas, eliminando filas, dentre outros. Na AAE foi implantado o Ambulatório de Especialidades com a tecnologia de atenção continua, realizada por equipe multiprofissional para hipertensos e diabéticos de alto e muito risco, estratificados na APS, compartilhando o cuidado. Uma das potencialidades da integração da APS e AAE foi o matriciamento realizado por profissionais da AAE, nas unidades laboratórios. A PAS configurou-se como um importante instrumento de gestão das RAS.

O Planejamento e a construção das Redes de Atenção à Saúde no DF, Brasil / Planning and building Health Care Networks in Brazil's Federal District

Resumo O artigo objetiva descrever o planejamento e a construção das Redes de Atenção à Saúde (RAS), por meio da Planificação da Atenção à Saúde (PAS), na Região Leste, Distrito Federal. Trata-se de um relato de experiência sobre a PAS, realizada de 2016 a 2018. As atividades foram desenvolvidas a partir de um conjunto de oficinas teóricas temáticas, de tutorias realizadas na Atenção Primária à Saúde (APS) e na Atenção Ambulatorial Especializada (AAE). Os resultados apontam uma melhor organização da RAS, especificamente para a linha de cuidado das condições crônicas, hipertensão e diabetes. Na APS foram organizados os macroprocessos: territorialização, cadastramento das famílias, estratificação de risco, classificação de riscos familiares, diagnóstico local, atendimento por bloco de horas, eliminando filas, dentre outros. Na AAE foi implantado o Ambulatório de Especialidades com a tecnologia de atenção continua, realizada por equipe multiprofissional para hipertensos e diabéticos de alto e muito risco, estratificados na APS, compartilhando o cuidado. Uma das potencialidades da integração da APS e AAE foi o matriciamento realizado por profissionais da AAE, nas unidades laboratórios. A PAS configurou-se como um importante instrumento de gestão das RAS.

Abstract This article reports on the experience of implementing Health Care Planning (HCP) in the territories of Itapoã, Paranoá and São Sebastião in the East Region of Brazil's Federal District. HCP began at the end of 2016 with Itapoã and was expanded to the other territories in 2018. The results point to a better organised health care network, specifically as regards care for chronic conditions, hypertension and diabetes. The activities involved a series of thematic theory workshops and tutoring workshops carried out in Primary Health Care (PHC) and Specialised Ambulatory Care (SAC) facilities. In PHC, macro-processes (territorialisation, family registration, risk stratification, family risk classification, local diagnosis, care by block of hours, elimination of waiting times, and others) were organised to support meeting certain of the population's demands. In SAC, an Ambulatory Specialities Clinic was set up using the technology of continuous care provided by a multi-professional team to high- and very high-risk hypertensive and diabetic patients stratified in PHC, and care provision is shared. One of the strong points in the integration of PHC and SAC was matrix support provided by SAC professionals in "laboratory units". HCP has been an important management tool for organising health care in the East Region.

A construção social da atenção primária à saúde / The social construction of primary health care

Este livro trata da construção, na prática social, da Atenção Primária à Saúde (APS). Portanto, há que se perguntar preliminarmente que APS se deseja construir? Essa pergunta faz sentido, tendo em vista que existem diferentes decodificaçõesde APS. A interpretação mais restrita da APS seletiva a entende como um programaespecífico destinado a populações e regiões pobres, às quais se oferta um conjunto restrito de tecnologias simples e de baixo custo, sem possibilidades de acesso a tecnologias de maior densidade. A interpretação da APS como o nível primário do sistema de atenção à saúde concebe-a como o modo de organizar e fazer funcionar a porta de entrada do sistema, enfatizando a função resolutiva desses serviços sobre os problemas de saúde mais comuns. E a interpretação mais ampla da APS como estratégia de organização do sistema de atenção à saúde que a compreende como uma forma singular de apropriar, recombinar, reorganizar e reordenar todos os recursos desse sistema para satisfazer às necessidades, demandas e representações da população, o que implica a inserção da APS em Redes de Atenção à Saúde (RAS). Há que se ressaltar que no SUS, ainda que o discurso oficial seja de APS como estratégia, na prática social essas três vertentes de interpretação dos cuidados primários se apresentam, coetaneamente, na prática social. É tempo de superar as duas primeiras interpretações e consolidar, definitivamente, a APS como a estratégia de organização do nosso sistema público de saúde. Isso implica superar o ciclo vigente da atenção básica em saúde caracterizado pela expansão do Programa de Saúde da Família (PSF) que não obstante seus bons resultados esgotou-se pela permanência de problemas estruturais que permanecem.

Astrocytes detect and upregulate transmission at inhibitory synapses of somatostatin interneurons onto pyramidal cells.

Astrocytes are important regulators of excitatory synaptic networks. However, astrocytes regulation of inhibitory synaptic systems remains ill defined. This is particularly relevant since GABAergic interneurons regulate the activity of excitatory cells and shape network function. To address this issue, we combined optogenetics and pharmacological approaches, two-photon confocal imaging and whole-cell recordings to specifically activate hippocampal somatostatin or paravalbumin-expressing interneurons (SOM-INs or PV-INs), while monitoring inhibitory synaptic currents in pyramidal cells and Ca2+ responses in astrocytes. We found that astrocytes detect SOM-IN synaptic activity via GABABR and GAT-3-dependent Ca2+ signaling mechanisms, the latter triggering the release of ATP. In turn, ATP is converted into adenosine, activating A1Rs and upregulating SOM-IN synaptic inhibition of pyramidal cells, but not PV-IN inhibition. Our findings uncover functional interactions between a specific subpopulation of interneurons, astrocytes and pyramidal cells, involved in positive feedback autoregulation of dendritic inhibition of pyramidal cells.

Oncostatin M promotes excitotoxicity by inhibiting glutamate uptake in astrocytes: implications in HIV-associated neurotoxicity.

BACKGROUND: Elevated levels of oncostatin M (OSM), an interleukin-6 cytokine family member, have been observed in HIV-1-associated neurocognitive disorders (HAND) and Alzheimer's disease. However, the function of OSM in these disease conditions is unclear. Since deficient glutamate uptake by astrocytes is instrumental in HAND-associated neurotoxicity, we hypothesized that OSM impairs glutamate uptake in astrocytes and thereby promotes neuronal excitotoxicity. METHODS: Primary cultures of mouse cortical astrocytes, neurons, microglia, and BV2 cell line were used. The expression of glutamate transporters (GLAST/EAAT1 and GLT-1/EAAT2) was investigated using real-time PCR and Western blot, and their activity was assessed by measuring (3)H-D-aspartate uptake. Neuronal toxicity was measured using the colorimetric MTT (3-(4,5-dimethylthiazol-2-yl-) 2,5-diphenyltetrazolium bromide) assay and immunocytochemistry. A chimeric HIV-1 that infects murine cells (EcoHIV/NL4-3-GFP virus (EcoHIV)) was used to investigate whether the virus induces OSM, OSM receptor (OSMR)-ß, glycoprotein 130 (gp130), GLT-1, GLAST (mRNA and protein), and OSM release (ELISA) in cultured BV2 cells, primary microglia, or astrocytes. Statistical analyses of the data were performed using one-way ANOVA (to allow multiple comparisons) and two-tailed Student's t test. RESULTS: OSM treatment (10 ng/mL) time-dependently reduced GLAST and GLT-1 expression and inhibited (3)H-D-aspartate uptake in cultured astrocytes in a concentration-dependent manner, an effect prevented by the Janus kinase (JAK)/signal transducers and activators of transcription (STAT)3 inhibitor AG490. Down-regulation of astrocytic glutamate transport by OSM resulted in NMDA receptor-dependent excitotoxicity in cortical neurons. Infection with EcoHIV induced OSM gene expression and protein release in BV2 cells and microglia, but not in astrocytes. Conversely, EcoHIV caused a fivefold increase in OSMR-ß mRNA (but not gp130) and protein in astrocytes, but not in microglia, which did not express OSMR-ß protein. Finally, astrocytic expression of GLAST gene was unaffected by EcoHIV, whereas GLT-1 mRNA was increased by twofold. CONCLUSIONS: We provide first evidence that activation of JAK/STAT3 signaling by OSM inhibits glutamate uptake in astrocytes, which results in neuronal excitotoxicity. Our findings with EcoHIV suggest that targeting OSMR-ß signaling in astrocytes might alleviate HIV-1-associated excitotoxicity.

Deletion of adenosine A2A receptors from astrocytes disrupts glutamate homeostasis leading to psychomotor and cognitive impairment: relevance to schizophrenia.

BACKGROUND: Adenosine A2A receptors (A2AR) modulate dopamine and glutamate signaling and thereby may influence some of the psychomotor and cognitive processes associated with schizophrenia. Because astroglial A2AR regulate the availability of glutamate, we hypothesized that they might play an unprecedented role in some of the processes leading to the development of schizophrenia, which we investigated using a mouse line with a selective deletion of A2AR in astrocytes (Gfa2-A2AR knockout [KO] mice]. METHODS: We examined Gfa2-A2AR KO mice for behaviors thought to recapitulate some features of schizophrenia, namely enhanced MK-801 psychomotor response (positive symptoms) and decreased working memory (cognitive symptoms). In addition, we probed for neurochemical alterations in the glutamatergic circuitry, evaluating glutamate uptake and release and the levels of key proteins defining glutamatergic signaling (glutamate transporter-I [GLT-I], N-methyl-D-aspartate receptors [NMDA-R] and α-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors [AMPA-R]) to provide a mechanistic understanding of the phenotype encountered. RESULTS: We show that Gfa2-A2AR KO mice exhibited enhanced MK-801 psychomotor response and decreased working memory; this was accompanied by a disruption of glutamate homeostasis characterized by aberrant GLT-I activity, increased presynaptic glutamate release, NMDA-R 2B subunit upregulation, and increased internalization of AMPA-R. Accordingly, selective GLT-I inhibition or blockade of GluR1/2 endocytosis prevented the psychomotor and cognitive phenotypes in Gfa2-A2AR KO mice, namely in the nucleus accumbens. CONCLUSIONS: These results show that the dysfunction of astrocytic A2AR, by controlling GLT-I activity, triggers an astrocyte-to-neuron wave of communication resulting in disrupted glutamate homeostasis, thought to underlie several endophenotypes relevant to schizophrenia.

Antagonistic interaction between adenosine A2A receptors and Na+/K+-ATPase-α2 controlling glutamate uptake in astrocytes.

Astrocytic glutamate transporter-1 (GLT-I) is critical to control the bulk of glutamate uptake and, thus, to regulate synaptic plasticity and excitotoxicity. GLT-I glutamate uptake is driven by the sodium gradient implemented by Na(+)/K(+)-ATPases (NKAs) and the α2 subunit of NKA (NKA-α2) is actually linked to GLT-I to regulate astrocytic glutamate transport. We recently found that adenosine A2A receptors (A2ARs), which control synaptic plasticity and neurodegeneration, regulate glutamate uptake through unknown mechanisms. Here we report that A2AR activation decreases NKA activity selectively in astrocytes to inhibit glutamate uptake. Furthermore, we found a physical association of A2ARs with NKA-α2s in astrocytes, as gauged by coimmunoprecipitation and in situ proximity ligation assays, in the cerebral cortex and striatum, two brain regions where A2ARs inhibit the astrocytic glutamate uptake. Moreover, the selective deletion of A2ARs in astrocytes (using Gfa2-A2AR-KO mice) leads to a concurrent increase of both astrocytic glutamate uptake and NKA-α2 levels and activity in the striatum and cortex. This coupling of astrocytic A2ARs to the regulation of glutamate transport through modulation of NKA-α2 activity provides a novel mechanism linking neuronal activity to ion homeostasis controlling glutamatergic activity, all of which are processes intricately associated with the etiology of several brain diseases.

Ecto-5'-nucleotidase (CD73)-mediated formation of adenosine is critical for the striatal adenosine A2A receptor functions.

Adenosine is a neuromodulator acting through inhibitory A1 receptors (A1Rs) and facilitatory A2ARs, which have similar affinities for adenosine. It has been shown that the activity of intracellular adenosine kinase preferentially controls the activation of A1Rs, but the source of the adenosine activating A2ARs is unknown. We now show that ecto-5'-nucleotidase (CD73), the major enzyme able to convert extracellular AMP into adenosine, colocalizes with A2ARs in the basal ganglia. In addition to astrocytes, striatal CD73 is prominently localized to postsynaptic sites. Notably, CD73 coimmunoprecipitated with A2ARs and proximity ligation assays confirmed the close proximity of CD73 and A2ARs in the striatum. Accordingly, the cAMP formation in synaptosomes as well as the hypolocomotion induced by a novel A2AR prodrug that requires CD73 metabolization to activate A2ARs were observed in wild-type mice, but not in CD73 knock-out (KO) mice or A2AR KO mice. Moreover, CD73 KO mice displayed increased working memory performance and a blunted amphetamine-induced sensitization, mimicking the phenotype of global or forebrain-A2AR KO mice, as well as upon pharmacological A2AR blockade. These results show that CD73-mediated formation of extracellular adenosine is responsible for the activation of striatal A2AR function. This study points to CD73 as a new target that can fine-tune A2AR activity, and a novel therapeutic target to manipulate A2AR-mediated control of striatal function and neurodegeneration.

Astrocytic adenosine A2A receptors control the amyloid-ß peptide-induced decrease of glutamate uptake.

Alzheimer's disease (AD) is characterized by a progressive cognitive impairment tightly correlated with the accumulation of amyloid-ß (Aß) peptides (mainly Aß(1-42)). There is a precocious disruption of glutamatergic synapses in AD, in line with an ability of Aß to decrease astrocytic glutamate uptake. Accumulating evidence indicates that caffeine prevents the burden of AD, likely through the antagonism of A(2A) receptors (A(2A)R) which attenuates Aß-induced memory impairment and synaptotoxicity. Since A(2A)R also modulate astrocytic glutamate uptake, we now tested if A(2A)R blockade could prevent the decrease of astrocytic glutamate uptake caused by Aß. In cultured astrocytes, Aß(1-42). (1 µM for 24 hours) triggered an astrogliosis typified by an increased density of GFAP, which was mimicked by the A(2A)R agonist, CGS 26180 (30 nM), and prevented by the A(2A)R antagonist, SCH 58261 (100 nM). Aß1-42 also decreased D-aspartate uptake by 28 ± 4%, an effect abrogated upon genetic inactivation or pharmacological blockade of A(2A)R. In accordance with the long term control of glutamate transporter expression by A(2A)R, Aß(1-42). enhanced the expression and density of astrocytic A(2A)R and decreased GLAST and GLT-I expression in astrocytes from wild type, but not from A(2A)R knockout mice. This impact of Aß(1-42). on glutamate transporters and uptake, dependent on A(2A)R function, was also confirmed in an ex vivo astrocyte preparation (gliosomes) from rats intracerebroventricularly (icv) injected with Aß(1-42). . These results provide the first demonstration for a direct key role of astrocytic A(2A)R in the ability of Aß-induced impairment of glutamate uptake, which may underlie glutamatergic synaptic dysfunction and excitotoxicity in AD.

Adenosine A2A receptors modulate glutamate uptake in cultured astrocytes and gliosomes.

Glutamate is the primary excitatory neurotransmitter in the central nervous system, where its toxic build-up leads to synaptic dysfunction and excitotoxic cell death that underlies many neurodegenerative diseases. Therefore, efforts have been made to understand the regulation of glutamate transporters, which are responsible for the clearance of extracellular glutamate. We now report that adenosine A(2A) receptors (A(2A) R) control the uptake of D-aspartate in primary cultured astrocytes as well as in an ex vivo preparation enriched in glial plasmalemmal vesicles (gliosomes) from adult rats, whereas A(1) R and A(3) R were devoid of effects. Thus, the acute exposure to the A(2A) R agonist, CGS 21680, inhibited glutamate uptake, an effect prevented by the A(2A) R antagonist, SCH 58261, and abbrogated in cultured astrocytes from A(2A) R knockout mice. Furthermore, the prolonged activation of A(2A) R lead to a cAMP/protein kinase A-dependent reduction of GLT-I and GLAST mRNA and protein levels, which leads to a sustained decrease of glutamate uptake. This dual mechanism of inhibition of glutamate transporters by astrocytic A(2A) R provides a novel candidate mechanism to understand the ability of A(2) (A) R to control synaptic plasticity and neurodegeneration, two conditions tightly associated with the control of extracellular glutamate levels by glutamate transporters.

Carboplatin and etoposide combined with bevacizumab for the treatment of recurrent glioblastoma multiforme.

Relapsed glioblastoma multiforme (GBM) responds poorly to standard therapies. Vascular endothelial growth factor (VEGF) is implicated in the development of GBM and the anti-VEGF monoclonal antibody bevacizumab has shown early clinical promise against malignant glioma. We treated six patients with recurrent GBM using bevacizumab combined with carboplatin and etoposide chemotherapy (ACE regimen). Toxicity was that expected for carboplatin and etoposide alone, except for an ischemic stroke in one patient. We observed partial responses in five patients and one responding patient developed extensive tumour necrosis after 2 cycles of treatment. Median progression-free and overall survival was 19 and 29.9weeks, respectively. Four responding patients developed recurrence, which was characterized by markedly less peri-tumoral edema, mass effect and necrosis compared with tumours at baseline. Two patients developed local extracranial extension. In conclusion, ACE was active in recurrent GBM and was mostly well tolerated.

Nesidioblastosis as a cause of focal pancreatic 111In-pentetreotide uptake in a patient with putative VIPoma: another differential diagnosis.

In adults, nesidioblastosis is a very infrequent condition and a rare cause of symptomatic presentations. The diagnosis of nesidioblastosis may be difficult with functional and anatomical imaging modalities. "Slight focal" pancreatic abnormalities using (111)In-pentetreotide imaging has been reported in patients with hyperinsulinaemic hypoglycaemia, confirmed histologically as nesidioblastosis. We describe a 60-year-old man who presented with a 1-year history of intermittent faecal urgency and refractory diarrhoea, non-specific laboratory results, negative imaging results (CT, MRI and EUS), a FNA biopsy that was inconclusive, but suggested an endocrine cell neoplasm, and a (111)In-pentetreotide scan that showed a moderately intense focal uptake clearly localised to the pancreatic head on a low-dose fusion CT. The histopathology of the specimen confirmed the diagnosis of nesidioblastosis.

Freud: as dimensöes do missivista; comunicaçäo e personalidade / Freud: dimensions of the letter writer; communication and personality

Procede-se a um exame panoramico da correspondencia ativa e passiva de Freud, em funcao do que foi ate o momento publicado em coletaneas especificamente de conteudo epistolografico. Trabalho confessadamente exploratorio e preliminar sobre o universo freudiano que se expressou eletivamente atraves de cartas a varios niveis de pessoas, distribuindo-se as mesmas, metodologicamente, nas categorias de cartas de carater geral (sociais, culturais, conviviais), cartas tecnico-cientificas (doutrinarias e intrumentos de comentarios e elaboracao, consolidacao e ampliacao da psicanalise e das ciencias correlatas) e cartas pessoais afetivas (basicamente as cartas de noivado). O autor acentua momentos significativos dessas varias especies de cartas, e finaliza seu trabalho apresentando extensa bibliografia de toda a materia tratada, com as indicacoes precisas de suas editoracoes. Ha, tambem, bibliografia indicativa das obras completas de Freud