Withdrawal induces distinct patterns of FosB/Delta FosB expression in outbred Swiss mice classified as susceptible and resistant to ethanol-induced locomotor sensitization

Chronic drug exposure and drug withdrawal induce expressive neuronal plasticity which could be considered as both functional and pathological responses. It is well established that neuronal plasticity in the limbic system plays a pivotal role in relapse as well as in compulsive characteristics of drug addiction. Although increases in FosB/DeltaFosB expression constitute one of the most important forms of neuronal plasticity in drug addiction, it is unclear whether they represent functional or pathological plasticity. It is of noteworthy importance the individual differences in the transition from recreational use to drug addiction. These differences have been reported in studies involving the ethanol-induced locomotor sensitization paradigm. in the present study we investigated whether sensitized and non-sensitized mice differ in terms of FosB/DeltaFosB expression. Adult male outbred Swiss mice were daily treated with ethanol or saline for 21 days. According to the locomotor activity in the acquisition phase, they were classified as sensitized (EtOH_High) or non-sensitized (EtOH_Low). After 18 h or 5 days, their brains were processed for FosB/DeltaFosB immunohistochemistry. On the 5th day of withdrawal, we could observe increased FosB/DeltaFosB expression in the EtOH_High group (in the motor cortex), in the EtOH_Low group (in the ventral tegmental area), and in both groups (in the striatum). Differences were more consistent in the EtOH_Low group. Therefore, behavioral variability observed in the acquisition phase of ethanol-induced locomotor sensitization was accompanied by differential neuronal plasticity during withdrawal period. Furthermore, distinct patterns of FosB/DeltaFosB expression detected in sensitized and non-sensitized mice seem to be more related to withdrawal period rather than to chronic drug exposure. Finally, increases in FosB/DeltaFosB expression during withdrawal period could be considered as being due to both functional and pathological plasticity. (C) 2013 Elsevier Inc. All rights reserved.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Psychiat & Med Psychol, BR-04038020 São Paulo, BrazilUniversidade Federal de São Paulo, Neurobiol Lab, BR-04039032 São Paulo, BrazilFac Med Sci Santa Casa São Paulo, Dept Physiol Sci, BR-01221020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Psychiat & Med Psychol, BR-04038020 São Paulo, BrazilUniversidade Federal de São Paulo, Neurobiol Lab, BR-04039032 São Paulo, BrazilWeb of Scienc

Atividades e expressão de peptidases em ratos tratados cronicamente com L-NAME, um inibidor da biossintese de oxido nitrico, e em ratos espontaneamente hipertensos

Orientador: Stephen HysloTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: As peptidases desempenham um papel importante na modulação da atividade de peptídeos endógenos em situações normais e patológicas. Neste trabalho, investigamos a atividade e expressão de quatro peptidases (aminopeptidase M- APM, dipeptidil peptidase IV - DPP IV, endopeptidase neutra - NEP 24.11 e metaloendopeptidase 24.15 - MEP 24.15) em tecidos (aorta, cérebro, coração, fígado, pulmão e rim) de ratos em dois modelos de hipertensão: a hipertensão induzida pelo tratamento crônico de ratos com NliJ-nitro-L-arginina metil éster (L-NAME), um inibidor da biossíntese de óxido nítrico (NO), e em ratos espontaneamente hipertensos (SHR). A atividade enzimática foi avaliada por ensaios enzimáticos fluorométricos ou colorimétricos enquanto a expressão foi avaliadapor westernblotting.Em ratos tratadoscom L-NAME(80 mg kg-1day-1, p.o., durante 4 semanas), não houve alteração na atividade das quatro peptidases estudas em cérebro, coração, fígado, pulmão e rim (a NEP 24.11 foi detectada apenas no pulmão e rim). Entretanto, na aorta torácica, a atividade da APM mostrou um pequeno decréscimo enquanto as atividades da DPP IV e MEP 24.15 foram significativamente aumentadas. A expressão de DPP IV e MEP 24.15 também foi aumentada pelo tratamento com L-NAME.As atividades da MEP 24.15 recombinante ou DPP IV renal não foram afetadas pela incubação com L-NAME (1-100 fJM)ou nitroprussiato de sódio e 3-morfolinosidnonimina (1-100 fJM cada), ambos doadores de NO. A administração de N-[1-(R,S)-carboxi-3-fenilpropil]-Ala- Aib-Tyr-p-aminobenzoato (JA2), um inibidor de MEP 24.15/MEP 24.16, em ratos anestesiados com pentobarbital sódico aumentou a resposta hipotensora da bradicinina. No modelo SHR, a atividade da DPP IV foi aumentada no cérebro, enquanto que as atividades da MEP 24.15 e NEP 24.11 foram aumentadas no pulmão (MEP 24.15 e NEP 24.11) e rim (NEP 24.11). Por outro lado, as atividades da DPP IV e MEP 24.15 foram reduzidas na aorta torácica; a APM foi inalterada e a NEP 24.11 não foi detectada neste tecido. Western blotting para DPP IV e MEP 24.15 mostrou redução correspondente na expressão de ambas as enzimas na aorta e um aumento para DPP IV no cérebro; não houve alteração na expressão da MEP 24.15 no pulmão e nem da NEP 24.11 no pulmão ou rim. A administração do JA2 em SHR não alterou as respostas à bradicinina. Estes resultados indicam que há alterações na atividade e expressão das peptidases estudadas (exceto APM) nos dois modelos de hipertensão. Entretanto, a variabilidade nestas alterações entre os dois modelos sugere que a hipertensão per se não é a causa principal destas diferenças, mas que provavelmente resultam de alterações bioquímicas associadas a cada modelo e seletivas para cada peptidaseAbstract: Peptidases play an important role in modulating the activity of endogenous peptides in normal and pathological situations. In this work, we investigated the activity and expression of four peptidases (aminopeptidase M - APM, dipeptidyl peptidase IV - DPP IV, metalloendopeptidase 24.15 - MEP 24.15 and neutral endopeptidase 24.11 - NEP 24.11) in rat tissues (aorta, brain, heart, kidney, liver and lung) in two models of hypertension, namely, chronic treatment with NlV-nitro-Larginine methyl ester (L-NAME), a nitric oxide (NO) synthase inhibitor (80 mg kg-1 dai1, p.o. for 4 weeks), and spontaneously hypertensive rats (SHR). Enzymatic activity was assayed fluorometrically or colorimetrically and protein expression was assessed by westem blotting. Treatment with L-NAME did not significantly alter the activities of the four peptidases in brain, heart, kidney, liver and lung (NEP was detected only in kidney and lung). In contrast, in thoracic aorta, the activity of APM was slightly but significantly reduced whereas those of DPP IV and MEP 24.15 were markedly enhanced. Immunoblotting for DPP IV and MEP 24.15 showed increased expression in aortic tissue. Neither L-NAME (1-100 J.lM)nor the NO donors sodium nitroprusside and 3-morpholinosydnonimine (1-100 J.lMeach) had any consistent effect on the activity of recombinant MEP 24.15 or renal DPP IV. Administration of the MEP 24.15/MEP 24.16 inhibitor N-[1-(R,S)-carboXy-3- phenylpropyl]-Ala-Aib-Tyr-p-aminobenzoate (JA2) in pentobartibal-anesthetized rats significantly potentiated the hypotensive response to bradykinin. In SHR, the activity of DPP IV was significantly increased in brain, whereas MEP 24.15 and NEP 24.11 activities were markedly enhanced in lung (MEP 24.15 and NEP 24.11) and kidney (NEP 24.11). In contrast, the activities of DPP IV and MEP 24.15 were markedly decreased in aortic tissue; APM was unaltered and NEP 24.11 was not detected in this tissue. Immunoblotting for DPP IV and MEP 24.15 showed decreased expression in aortic tissue and increased expression of DPP IV in the brain; there was no alteration in the expression of MEP 24.15 in the lung or of NEP 24.11 in kidney and lung. The administration of JA2 to SHR did not alter the responses to bradykinin. These results indicate that there were alterations in the activity and expression of the peptidases studied (except for APM) in both models of hypertension. However, the different patterns of afterations seen between the two models suggests that the changes were not caused by the hypertension per se, but rather were probably the result of biochemical alterations associated with each model and selective for each peptidaseDoutoradoDoutor em Farmacologi

Participação de fibras sensoriais na resposta edematogenica induzida pela enterotoxina estafilococica do tipo B em camundongos

Orientador: Edson AntunesObservação: Faltam as páginas II, IV, IX, X, 61 e 67Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Neste trabalho, investigou-se a participação de fibras sensonaIS no aumento de penneabilidade vascular e na fonnação de edema induzidos pela SEB (enterotoxina estafilocócica do tipo B) (25 ~g/pata) em camundongos. O tratamento dos animais com o SR140333 (antagonista de receptores NK1; 120 nmoles/Kg, s.c. + 120 nmoles/Kg, i.v.) inibiu significativaInente o edema e a exsudação (35 e 37%, respectivamente) induzidos pela SEB. Da mesma fonna, o edema e a exsudação plasmática induzidos pelo GR73632 (agonista de receptores NK1; 50 pmoles/pata) foram significativamente reduzidos pelo SR140333 (40 e 88%, respectivamente). Ao contrário, o tratamento endovenoso cQm SR48968 (antagonista de receptores NK2; 1,7 J..U11oles/Kg) não alterou o edema e a exsudação causados pela SEB (ou GR73632) em relação aos seus respectivos controles. O edema e a exsudação plasmática induzidos pela SEB não se alteraram, significativamente, pela co-injeção de CGRP (100 e 300 pmoles/pata). Por outro lado, o edema evocado pelo GR73632 (50 pmoles/pata) foi significativamente potencializado pelo CGRP em ambas as doses. No grupo de animais tratados com o HOE 140 (antagonista de receptores B2; 400 mnoles/Kg, i.v.) observou-se redução significativa do edema e da exsudação plasmática induzidos pela SEB (35 e 34%, respectivaInente). Da mesma fonna, o edema e a exsudação plasmática evocados pela bradicinina (3 mnoles/pata) foram significativamente reduzidos (69 e 94%, respectivamente) pelo tratamento com o HOE 140. O trataInento prévio dos animais com a mistura, de antagonistas, SR140333 (120 nrnoles/Kg; s.c. + 120 nrnoles/Kg; i.v.) e HOE 140 (400 l1l1}.oles/Kg; i.v.), reduziu significativamente o edema e a exsudação evocados pela SEB. Entretanto, a inibição, embora signi:ticativa, não foi maior do que aquela observada pelo SR140333 ou HOE 140, administrados isoladamente. Além disso, o SR140333 reduziu significativamente o edema (62%) e a exsudação (49%) evocados pela bradicinina (3 nrnoles/pata). o edema e a exsudação plasmática causados pela SEB no grupo tratado com a capsazepina (antagonista de receptores vanilóides; 100 /lmoles/Kg; s.c.) foi significativamente menor (53 e 45%, respectivamente), em relação ao grupo controle. A capsazepina (50 e 100 /lmoles/Kg; s.c.) reduziu, em ambas as doses, o edema induzido p~la capsaicina (5 /lg/pata), em relação ao grupo controle (50 e 67 %, respectivamente). No grupo tratado com mepiramina (antagonista de receptores HI; 10 mg/Kg, i.v.) ou metisergida (antagonista de receptores 5-HT; 10 mg/Kg, i.v.), observou-se que o edema evocado pela SEB foi reduzido significativamente, quando comparado aos respectivos. grupos-controle ( 40 e 60%, respectivamente). A exsudação plasmática nos animais tratados cqin mepiramina e metisergida foi reduzida em 25 e 63%, respectivamente. O tratamento com mepiramina ou metisergida reduziu o edema e a exsudação induzidos tanto pela histamina (100 /lmoles/pata), quanto pela serotonÍna (100 /lmoles/pata), respectivamente. Em animais diabéticos, observou-se que o edema e a exsudação plasmática -induzidos pela SEB estavam reduzidos, em relação aos animais não diabéticos (75 e 50%, respectivamente); o mesmo foi observado com o edema e com a exsudação plasmática evocados pela capsaicina (3,2 /lg/pata) (56 e 77%, respectivamente). O tratamento prévio dos animais com insulina (20 UIlKg; s.c.) corrigiu a hipergl(cemia, mas não promoveu alterações significativas na resposta inflamatória induzida pela SEB ou capsaicina. Em resumo, nossos resultados mostraram que a SEB causa uma resposta inflamatória neurogênica complexa, que envolve geração local de cininas, ativação de receptores vanilóides e degranulação de mastócitosAbstract: In this study, we investigated the participation of sensory fibers in the mouse paw oedema induced by staphylococcãI enterotoxin type B (SEB; 25 Ilg/paw). The NK1 receptor antagonist SR140333 (120 nmolJkg, s.c. + 120 nmolJkg, i.v.) significantly inhibited the plasma protein extravasation and oedema formation induced by SEB (37 and 35%, respectively). The oedema and exsudation induced by the NK1 receptor agonist (GR73632; 50 pmol/paw) were also reduced significantly by SR140333 (40 and 88%, respectively). The oedematogenic activity of the NK2 receptor antagonist SR48968 (1.7 J.l11lolJKg, i.v.) did not affect SEB or GR73632. In addition, the oedema and plasma exsudation-induced by SEB were not altered by the vasodilator CGRP (100 and 300 pmol/paw). However, GR73632-induced oedema (50 pmol/paw) was significantly increased by CGRP. Pretreating the mice with the B2 antagonist receptor (HOE 140; 400 nmolJk~, i.v.) showed a significantly reduced the SEB-induced oedema and exsudation (35 and 34%, respectively). Oedema caused by bradykinin (3 nmol/paw) was also significantly reduced by HOE 140. Simultaneol!J,s treatment with SR140333 and HOE 140 inhibited SEB-induced paw oedem~, but only to an extent similar to that observed when these antagonists were administered alone. SR140333 (120 nmolJkg, s.c. + 120 nmolJkg, i.v.) significantly reduced the oedema (62%) and exsudation (49%) induced by bradykinin. The vanilloid antagonist capsazepine (100 JlmolJKg, s.c.) significantly reduced in the oedema (53%) and exsudation (45%) induced by SEB. This antagonist (50 and 100 JlmolJkg, s.c.) also dose-dependently inhibited the paw oedema induced by capsaicin (5 Ilg/paw)MestradoMestre em Farmacologi

The use of aromatase inhibitors in boys with short stature: what to know before prescribing?

ABSTRACT Aromatase is a cytochrome P450 enzyme (CYP19A1 isoform) able to catalyze the conversion of androgens to estrogens. The aromatase gene mutations highlighted the action of estrogen as one of the main regulators of bone maturation and closure of bone plate. The use of aromatase inhibitors (AI) in boys with short stature has showed its capability to improve the predicted final height. Anastrozole (ANZ) and letrozole (LTZ) are nonsteroidal inhibitors able to bind reversibly to the heme group of cytochrome P450. In this review, we describe the pharmacokinetic profile of both drugs, discussing possible drug interactions between ANZ and LTZ with other drugs. AIs are triazolic compounds that can induce or suppress cytochrome P450 enzymes, interfering with metabolism of other compounds. Hydroxilation, N-dealkylation and glucoronidation are involved in the metabolism of AIs. Drug interactions can occur with azole antifungals, such as ketoconazole, by inhibiting CYP3A4 and by reducing the clearance of AIs. Antiepileptic drugs (lamotrigine, phenobarbital, and phenytoin) also inhibit aromatase. Concomitant use of phenobarbital or valproate has a synergistic effect on aromatase inhibition. Therefore, it is important to understand the pharmacokinetics of AIs, recognizing and avoiding possible drug interactions and offering a safer prescription profile of this class of aromatase inhibitors. Arch Endocrinol Metab. 2017;61(3):391-7

Peptidase Activities In Rats Treated Chronically With N(omega)-nitro-l-arginine Methyl Ester (l-name).

The chronic treatment of rats with N(omega)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) biosynthesis, results in hypertension. This inhibition of NO production results in activation of the renin-angiotensin system, with increased activity of the carboxypeptidase angiotensin I-converting enzyme (ACE). Since chronic NO inhibition increases ACE activity, we hypothesized that this inhibition could also affect the activities of other peptidases involved in cardiovascular functions. To test this possibility, we examined the activities of aminopeptidase M (APM), dipeptidyl peptidase IV (DPP IV), metalloendopeptidase 24.15 (MEP 24.15) and neutral endopeptidase 24.11 (NEP 24.11) in rat brain, heart, kidney, liver, lung and thoracic aorta. Male Wistar rats were treated chronically with L-NAME (80mgkg(-1) per day) administered in the drinking water for 4 weeks and their organs then removed and processed for the determination of peptidase activities. Treatment with L-NAME did not significantly alter the activities of the four peptidases in brain, heart, kidney, liver and lung. In contrast, in aorta, the activity of APM was slightly but significantly reduced whereas those of DPP IV and MEP 24.15 were markedly enhanced; NEP 24.11 was not detected in this tissue. Immunoblotting for DPP IV and MEP 24.15 showed increased expression in aortic tissue. Neither L-NAME (1-100microM) nor the NO donors sodium nitroprusside and 3-morpholinosydnonimine (SIN-1; 1-100microM) had any consistent effect on the activity of recombinant MEP 24.15 or renal DPP IV. The importance of MEP 24.15 in peptide metabolism was confirmed in pentobartibal-anesthetized rats pretreated with the MEP 24.15 inhibitor N-[1-(R,S)-carboxy-3-phenylpropyl]-Ala-Aib-Tyr-p-aminobenzoate (JA2), which significantly potentiated the hypotensive response to bradykinin. The altered peptidase activities seen in aorta may contribute to modulating vascular responses in this model of hypertension.68205-1

Ocorrência de Rhopalopsyllus lutzi lutzi (Baker) (Siphonaptera, Rhopalopsyllidae) em Canis familiaris (Linnaeus) de zona rural do município de Piraí, Rio de Janeiro, Brasil Occurrence of Rhopalopsyllus lutzi lutzi (Baker) (Siphonaptera, Rhopalopsyllidae) on Canis familiaris (Linnaeus) from rural areas of the county of Piraí, State of Rio de Janeiro, Brazil

A ocorrência de Rhopalopsyllus lutzi lutzi (Baker) (Siphonaptera, Rhopalopsyllidae) foi assinalada em Canis familiaris (Linnaeus) de áreas rurais do município de Piraí, estado do Rio de Janeiro, Brasil. No período de junho 2001/novembro 2003, 51 sifonápteros foram capturados em oito cães procedentes de duas propriedades rurais do município de Piraí. Os exemplares coletados foram acondicionados em álcool etílico 70%, levados ao Laboratório de Parasitologia Animal da Universidade Federal Rural do Rio de Janeiro para contagem e sexagem. Os exemplares foram identificados como Rhopalopsyllus lutzi lutzi Baker, 1904 (machos=18 e fêmeas=33). R. lutzi lutzi é, pela primeira vez, assinalada em cães domésticos naturalmente infestados em áreas rurais do estado do Rio de Janeiro.<br>From June/2001 to November/2003, 51 fleas were collected on eight dogs from two rural properties of the county of Piraí, State of Rio de Janeiro. After preservation in 70% ethanol, the fleas were counted and sexed in the Laboratório de Parasitologia Animal da Universidade Federal Rural do Rio de Janeiro. The samples were identificated as Rhopalopsyllus lutzi lutzi (Baker, 1904) (male=18 and female=33). For the first time R. lutzi lutzi was found paraziting dogs in rural areas of the State of Rio de Janeiro