This dissertation consists of three essays in health economics. The three chapters focus specifically on prescription drug use and treatment in various national and state settings and evaluate the impact of government policies and interventions on this sector of the health care market. The first two chapters focus on opioid prescribing in the United States. Prescription drug monitoring programs (PDMPs or PMPs)—online systems that health care providers and pharmacists can use to query patient prescription records—are one of the most widely-used state tools in regulating the prescribing and dispensing of opioids. However, the staggered adoption of PDMPs over time has created opportunities for patients to evade monitoring by going to a state that does not have a PDMP. Chapter 1 evaluates how spillovers attributable to policy non-coordination between neighboring states impact the effectiveness of PDMPs. I find that after prescribers gain access to PDMPs, opioid volume and prescription opioid deaths decrease in counties with a PDMP that are insulated from opportunities for evasion. I find a similar effect in counties with a PDMP that are exposed to evasion. This suggests that exposure to evasion through proximity to non-PDMP areas does not significantly attenuate the policy effect. I also find evidence that opioid volume and prescription opioid deaths decrease in counties without a PDMP that are exposed to spillovers from counties with the policy. Illicit opioid deaths are not affected in any counties with a PDMP but decrease in counties without a PDMP that are exposed to spillovers. I discuss the potential mechanisms through which spillovers may operate. Chapter 2, which is joint work with Adam Sacarny, David Powell, Ian Williamson, Weston Merrick, and Mireille Jacobson, evaluates how "nudge" interventions can impact the behavior of clinicians prescribing controlled substances. PMPs aim to reduce inappropriate opioid prescribing but may be underutilized by prescribers. We conduct a randomized clinical trial of 12,000 clinicians in Minnesota to test whether letters to providers can increase PMP use and decrease potentially dangerous opioid co-prescriptions. In this study, we focus on the co-prescribing of opioids and benzodiazepines and the co-prescribing of opioids and gabapentinoids. We find that letters that mention the state's new PMP use mandate increase PMP search rates and the share of clinicians with PMP accounts but have no significant effect on co-prescribing. Letters with only information about the risks of co-prescribing and a list of co-prescribed patients have no detected effect on primary outcomes of interest. We also explore the impact of the letters on additional search and prescribing outcomes. Our results highlight the potential for simple letter-based interventions to encourage engagement with PMPs and facilitate better-informed prescribing of opioids and other medications. Finally, Chapter 3 studies the prescription drug market in Japan, examining how changes in health care prices faced by patients can influence demand. I exploit a feature of the Japanese healthcare system, where an individual's coinsurance rate is determined primarily by their age, to evaluate the impact of a change in patient cost sharing on total prescription drug spending. I contribute to the existing literature by investigating heterogeneous effects by patient sex and drug therapeutic class (focusing on cardiovascular drugs, antibiotics, vitamins, antihistamines, and psychotropic drugs). I find that for the whole sample, price elasticity of spending for prescription drugs is comparable to previous estimates of price elasticity of spending for general medical services. I find no evidence of heterogeneous effects by sex over the whole sample of prescriptions, but I do find statistically significant differences between women and men within therapeutic drug classes. I also conduct exploratory analysis on the effect of changes in patient cost sharing on prescription drug volume. I estimate a price elasticity of demand for prescription drugs that is larger than previous estimates of demand elasticity for general medical services. I also find evidence that physicians do not respond on the intensive margin by prescribing more expensive medications. Although Japanese patients are more likely to be prescribed brand-name drugs, patients using generic medications may be more price sensitive to changes in patient cost sharing.

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Yes / Designer benzodiazepines are one of the primary new psychoactive substances (NPS) threats around the world, being found in large numbers in post-mortem, driving under the influence of drugs (DUID) and drug facilitated sexual assault (DFSA) cases. Even though when compared to many other NPS types, such as opioids and cathinones, there are relatively few designer benzodiazepines being monitored. Recently a new NPS benzodiazepine has been reported in Europe, the USA and Canada, desalkygidazepam, also known as bromonordiazepam. This substance is a metabolite of the pro-drug gidazepam, a drug licenced for use in Ukraine and Russia under the name Gidazepam IC®. In the paper we review what is currently known about the use, pharmacology and analytical detection of gidazepam, its metabolite desalkygidazepam, and their other possible metabolites.

INTRODUÇÃO: O midazolam é uma imidazobenzodiazepina usada para induzir o sono, produzir sedação antes de procedimentos dolorosos e no tratamento do estado de mal epiléptico. Seu uso pela via intranasal proporciona um rápido início de ação e esta via, em muitos casos, pode substituir as vias endovenosa e intramuscular, mais invasivas. Assim, o midazolam intranasal tem sido sugerido no tratamento extra-hospitalar de crises epilépticas e ataques de pânico. Por outro lado, os benzodiazepínicos possuem um potencial para serem abusados, principalmente em usuários de outras drogas. OBJETIVO: o presente estudo objetivou verificar o potencial de abuso do midazolam intranasal numa população experiente no uso intranasal de substâncias - abusadores de cocaína aspirada. MÉTODOS: Foram estudados 31 voluntários abusadores ou dependentes de cocaína e 34 controles saudáveis, subdivididos em quatro grupos: Abusadores de Cocaína (N = 16) e Voluntários Saudáveis (N= 17) que receberam midazolam (0,5 mg de hidrocloreto de midazolam em cada narina), e Abusadores de Cocaína (N = 15) e Voluntários Saudáveis (N = 17) que receberam o mesmo volume de um placebo ativo. As variáveis de resposta foram a Apreciação da Substância (AS) e a Vontade de Repetir o Uso da Substância (VR), avaliadas através de escalas analógicas visuais. RESULTADOS: A análise de perfis para medidas repetidas das variáveis de resposta mostrou um efeito significante da variável Tempo sobre AS (F[5;57] =3,947, p=0,004) e VR (F[5;57] =3,311, p=0,011). A variável Grupo (Abusadores de Cocaína x Voluntários Saudáveis) também teve um impacto sobre as variáveis de resposta AS e VR, sendo que os Abusadores de Cocaína tiveram pontuações mais altas tanto em AS (F[5;57] = 4,946, p = 0,030) quanto em VR (F[5;57] =5,229, p=0,026). Numa análise de regressão linear para investigar os efeitos do humor - medidos através de uma Escala Visual Analógica do Humor (VAMS) - sobre as variáveis de resposta AS e VR, os Abusadores de Cocaína apresentaram escores maiores que os Voluntários saudáveis tanto para AS (t = 3,37; p = 0,001) quanto para VR (t = 5,607; p = 0,011). Observou-se, também, um efeito dos fatores VAMS 16 (MAIS DEPRIMIDO MAIS EUFÓRICO; t = 4,28; p < 0,001) e VAMS 12 (MAIS EXCITADO MAIS RELAXADO; t = 2,66; p = 0,010), sobre a variável de resposta AS (R2 = 0,32): níveis maiores de euforia e relaxamento predisseram uma maior Apreciação da Substância instilada. O fator VAMS 16 (MAIS DEPRIMIDO MAIS EUFÓRICO) teve um efeito também sobre a variável de resposta VR (t = 3,65, p < 0,001; R2 =0,24): maior euforia predisse maior vontade de repetir o uso da substância. Finalmente, uma análise de regressão linear utilizando-se AS como variável explicativa e VR como variável de resposta resultou que quanto maior a apreciação positiva da substância, maior era a vontade de repetir seu uso (F = 108, 517; p < 0,001; R2 = 0,65). CONCLUSÕES: Corroborando estudos anteriores, observou-se que sensações como relaxamento e euforia correlacionam-se com o potencial de abuso de uma substância e uma maior apreciação positiva dos efeitos de uma substância correlaciona-se com uma maior vontade de usá-la novamente. Por outro lado, conclui-se que via intranasal em si aumentaria a probabilidade de abuso em usuários de substâncias intranasais. Consequentemente, uma eventual produção e comercialização para uso intranasal de uma substância com potencial de abuso deverá levar em conta este risco adicional para populações usuárias de drogas / INTRODUCTION: Midazolam is an imidazobenzodiazepine used for sleep induction, for sedation before painful procedures and in the treatment of status epilepticus. When it is administered through the intranasal route, it has a fast beginning action and this route can many times be substituted for the more invasive intravenous and intramuscular routes. Therefore, intranasal midazolam has been uggested for the community management of pileptic seizures and panic attacks. On the other side, benzodiazepines display abuse liability, particularly in substance abusers. OBJECTIVE: The present study aimed at examining the abuse liability of intranasal midazolam in a population experienced with the intranasal abuse of substances, namely snorted cocaine abusers. METHODS: Thirty-one subjects with diagnoses of snorted-cocaine abuse or dependence have been studied, divided in four groups: Cocaine-Abusers (n = 16) and Healthy Volunteers (n = 17) that received midazolam (0.5 mg midazolam hydrochloride in each nostril), and Cocaine-Abusers (n = 15) and Healthy Volunteers (n = 17) that received the same volume of an active placebo. Response variables were Substance Liking (SL) and the Desire to Take the Substance Again (SA), assessed through visual analogue scales. RESULTS: Profile analysis for repeated measures of the response variables showed a significant effect of Time over both SL (F[5,57] =3.947, p=0.004) and SA (F[5;57] =3.311, p=0.011). Group had also a significant effect, in that Cocaine Abusers scored higher in both SL (F[5,57] = 4.946, p = 0.030) and SA (F[5;57] =5.229, p=0.026). In a linear regression analysis examining the effects of mood (measured through Visual Analogic Mood Scales VAMS) over the response variables SL and SA, Cocaine Abusers displayed higher scores than Healthy Volunteers at both SL (t = 3.37; p = 0.01) and SA (t = 5.607; p = 0.011). It was also found that variables VAMS 16 (MORE DEPRESSED MORE EUPHORIC; t = 4.28; p < 0.001) and VAMS 12 (MORE EXCITED MORE RELAXED; t = 2.66; p = 0.010) had an effect over response variable SL (R2 = 0.32): higher euphoria and relaxation scores predicted more liking of the administered substance. Factor VAMS 16 (MORE DEPRESSED MORE EUPHORIC) had also an effect over response variable SA (t = 3.65, p < 0.001; R2 =0.24): more euphoria predicted more desire to take the drug again. Finally, in a linear regression with SL as explaining variable and SA as response variable, it was found that higher SL predicted a higher SA (F = 108.517; p < 0.001; R2 = 0.65). CONCLUSIONS: Corroborating previous findings in literature, it was observed that feelings of relaxation and euphoria after the administration of an intranasal substance are correlated with higher abuse liability and that subjects who report more liking of a substance do also report more desire to take it again. On the other hand, it is concluded that the intranasal route might per se increase the probability of abuse in intranasal-substance users. Therefore, the production and marketing for intranasal use of a substance with abuse liability should take into account this additional risk for intranasal drug abusing populations.

INTRODUÇÃO: O midazolam é uma imidazobenzodiazepina usada para induzir o sono, produzir sedação antes de procedimentos dolorosos e no tratamento do estado de mal epiléptico. Seu uso pela via intranasal proporciona um rápido início de ação e esta via, em muitos casos, pode substituir as vias endovenosa e intramuscular, mais invasivas. Assim, o midazolam intranasal tem sido sugerido no tratamento extra-hospitalar de crises epilépticas e ataques de pânico. Por outro lado, os benzodiazepínicos possuem um potencial para serem abusados, principalmente em usuários de outras drogas. OBJETIVO: o presente estudo objetivou verificar o potencial de abuso do midazolam intranasal numa população experiente no uso intranasal de substâncias - abusadores de cocaína aspirada. MÉTODOS: Foram estudados 31 voluntários abusadores ou dependentes de cocaína e 34 controles saudáveis, subdivididos em quatro grupos: Abusadores de Cocaína (N = 16) e Voluntários Saudáveis (N= 17) que receberam midazolam (0,5 mg de hidrocloreto de midazolam em cada narina), e Abusadores de Cocaína (N = 15) e Voluntários Saudáveis (N = 17) que receberam o mesmo volume de um placebo ativo. As variáveis de resposta foram a Apreciação da Substância (AS) e a Vontade de Repetir o Uso da Substância (VR), avaliadas através de escalas analógicas visuais. RESULTADOS: A análise de perfis para medidas repetidas das variáveis de resposta mostrou um efeito significante da variável Tempo sobre AS (F[5;57] =3,947, p=0,004) e VR (F[5;57] =3,311, p=0,011). A variável Grupo (Abusadores de Cocaína x Voluntários Saudáveis) também teve um impacto sobre as variáveis de resposta AS e VR, sendo que os Abusadores de Cocaína tiveram pontuações mais altas tanto em AS (F[5;57] = 4,946, p = 0,030) quanto em VR (F[5;57] =5,229, p=0,026). Numa análise de regressão linear para investigar os efeitos do humor - medidos através de uma Escala Visual Analógica do Humor (VAMS) - sobre as variáveis de resposta AS e VR, os Abusadores de Cocaína apresentaram escores maiores que os Voluntários saudáveis tanto para AS (t = 3,37; p = 0,001) quanto para VR (t = 5,607; p = 0,011). Observou-se, também, um efeito dos fatores VAMS 16 (MAIS DEPRIMIDO MAIS EUFÓRICO; t = 4,28; p < 0,001) e VAMS 12 (MAIS EXCITADO MAIS RELAXADO; t = 2,66; p = 0,010), sobre a variável de resposta AS (R2 = 0,32): níveis maiores de euforia e relaxamento predisseram uma maior Apreciação da Substância instilada. O fator VAMS 16 (MAIS DEPRIMIDO MAIS EUFÓRICO) teve um efeito também sobre a variável de resposta VR (t = 3,65, p < 0,001; R2 =0,24): maior euforia predisse maior vontade de repetir o uso da substância. Finalmente, uma análise de regressão linear utilizando-se AS como variável explicativa e VR como variável de resposta resultou que quanto maior a apreciação positiva da substância, maior era a vontade de repetir seu uso (F = 108, 517; p < 0,001; R2 = 0,65). CONCLUSÕES: Corroborando estudos anteriores, observou-se que sensações como relaxamento e euforia correlacionam-se com o potencial de abuso de uma substância e uma maior apreciação positiva dos efeitos de uma substância correlaciona-se com uma maior vontade de usá-la novamente. Por outro lado, conclui-se que via intranasal em si aumentaria a probabilidade de abuso em usuários de substâncias intranasais. Consequentemente, uma eventual produção e comercialização para uso intranasal de uma substância com potencial de abuso deverá levar em conta este risco adicional para populações usuárias de drogas / INTRODUCTION: Midazolam is an imidazobenzodiazepine used for sleep induction, for sedation before painful procedures and in the treatment of status epilepticus. When it is administered through the intranasal route, it has a fast beginning action and this route can many times be substituted for the more invasive intravenous and intramuscular routes. Therefore, intranasal midazolam has been uggested for the community management of pileptic seizures and panic attacks. On the other side, benzodiazepines display abuse liability, particularly in substance abusers. OBJECTIVE: The present study aimed at examining the abuse liability of intranasal midazolam in a population experienced with the intranasal abuse of substances, namely snorted cocaine abusers. METHODS: Thirty-one subjects with diagnoses of snorted-cocaine abuse or dependence have been studied, divided in four groups: Cocaine-Abusers (n = 16) and Healthy Volunteers (n = 17) that received midazolam (0.5 mg midazolam hydrochloride in each nostril), and Cocaine-Abusers (n = 15) and Healthy Volunteers (n = 17) that received the same volume of an active placebo. Response variables were Substance Liking (SL) and the Desire to Take the Substance Again (SA), assessed through visual analogue scales. RESULTS: Profile analysis for repeated measures of the response variables showed a significant effect of Time over both SL (F[5,57] =3.947, p=0.004) and SA (F[5;57] =3.311, p=0.011). Group had also a significant effect, in that Cocaine Abusers scored higher in both SL (F[5,57] = 4.946, p = 0.030) and SA (F[5;57] =5.229, p=0.026). In a linear regression analysis examining the effects of mood (measured through Visual Analogic Mood Scales VAMS) over the response variables SL and SA, Cocaine Abusers displayed higher scores than Healthy Volunteers at both SL (t = 3.37; p = 0.01) and SA (t = 5.607; p = 0.011). It was also found that variables VAMS 16 (MORE DEPRESSED MORE EUPHORIC; t = 4.28; p < 0.001) and VAMS 12 (MORE EXCITED MORE RELAXED; t = 2.66; p = 0.010) had an effect over response variable SL (R2 = 0.32): higher euphoria and relaxation scores predicted more liking of the administered substance. Factor VAMS 16 (MORE DEPRESSED MORE EUPHORIC) had also an effect over response variable SA (t = 3.65, p < 0.001; R2 =0.24): more euphoria predicted more desire to take the drug again. Finally, in a linear regression with SL as explaining variable and SA as response variable, it was found that higher SL predicted a higher SA (F = 108.517; p < 0.001; R2 = 0.65). CONCLUSIONS: Corroborating previous findings in literature, it was observed that feelings of relaxation and euphoria after the administration of an intranasal substance are correlated with higher abuse liability and that subjects who report more liking of a substance do also report more desire to take it again. On the other hand, it is concluded that the intranasal route might per se increase the probability of abuse in intranasal-substance users. Therefore, the production and marketing for intranasal use of a substance with abuse liability should take into account this additional risk for intranasal drug abusing populations.

Abstract, key words The aim of this thesis was to perform a phytoextraction experiment with benzodiazepines chlordiazepoxide, diazepam, alprazolam and bromazepam on corn plant (Zea mays). After 14 days of growing of sterile cultivation, new medium (Murashige and Skoog) contaminated with benzodiazepine was added. The starting concentration of benzodiazepine was 10 mg · l-1. After every 24 hours, a sample of medium was collected. The actual concentration of benzodiazepine was measured on HPLC with UV detection. Extractable residues were also analysed to find out whether the benzodiazepine is being translocated to the upper parts of the plant. The same HPLC conditions were used for these samples. The greatest phytoextraction efficiency (the amount of drug extracted by 1 gram of biomass in 24 hours) was observed for chlordiazepoxide, followed by bromazepam, alprazolam and diazepam respectively. The extractable residues analysis confirmed the translocation to the upper parts of the plant for every of the benzodiazepines tested. That indicates a threat for the animals through the food chain contamination. Key words: phytoremediation, phytoextraction, benzodiazepines, extractable residuals, HPLC.

O objetivo do presente estudo foi determinar a prevalência do uso de benzodiazepínicos (BZDs) em mulheres de uma unidade da Estratégia de Saúde da Família (ESF). Analisou-se também as possíveis associações do uso de BZD com fatores sociodemográficos, acometimento de doenças crônicas e uso de outros psicotrópicos. O recorte de gênero justifica-se pelas evidências científicas de maior uso destes fármacos por mulheres. O lócus da atenção básica foi escolhido a partir da observação sobre a carência de estudos, especialmente brasileiros, neste setting. Trata-se de um estudo quantitativo, de corte transversal e caráter correlacional e descritivo. Os dados foram coletados nos prontuários, fichas cadastrais e arquivo de receitas da farmácia responsável pela região. Foram realizadas análises através de estatística descritiva, análises bivariadas e regressão logística considerando como variável desfecho o uso de BZDs. A prevalência encontrada no presente estudo foi de 7,4% (n=1094), abaixo das descritas em estudos similares. Entende-se que o foco mais preventivo das ESFs possam contribuir para estes achados ou que a qualidade dos registros tenha interferido nestes resultados. As usuárias de BZDs estudadas estavam em sua maioria na faixa de idade de 56 a 74 anos (48,4%), estudaram somente até o primeiro grau completo (65,6%), autorreferiram pelo menos uma doença crônica (82,2%) e quase metade usava, concomitantemente, pelo menos um outro psicotrópico (48,4%). Houve associação significativa entre usar BZDs e ter baixa escolaridade (p=0,005), apresentar doenças autorreferidas (p=0,000) e consumir outros psicotrópicos (p=0,000). A associação entre idades mais elevadas e uso de BZDs (p=0,005) também foi significativa. As análises apontaram ainda que as mulheres que usavam outros psicotrópicos apresentaram cinco vezes mais chances de usar BZDs do que as que não usavam. As mulheres com doença autorreferida apresentaram quase cinco vezes mais chances de usarem BZDs do que as que não referiram doenças. Conclui-se que o grupo que deve ser priorizado em relação ao consumo destas substâncias é o de mulheres de meia idade ou mais, com pouca escolaridade e doença crônica. Recomenda-se que as equipes se atentem para as necessidades psicossociais das pessoas com transtorno mental, discutam e empreendam esforços para implementar ações para inserção e reinserção social e de ampliação das redes de apoio que possam ser estratégicas no auxílio à contenção dos sintomas e melhoria da qualidade de vida dessas pessoas, na tentativa de evitar o uso de mais psicotrópicos e/ou de serviços de maior complexidade / The aim of the present study was identifying the prevalence of Benzodiazepine (BZD) by women in a Family health care program (FHPs). It was also analyzed the possible associations of the use of BZD with sociodemographic factors, affection of chronic diseases and use of other psychotropic drugs. The choice of gender was justified by scientific literature pointing women to be the major BZD users. The choice of basic health care was done because of the lack in studies in those settings in Brazil. It is a quantitative study of cross section and of correlational descriptive type. Data were collected on the charts and registration sheets and recipes from responsible pharmacy for the geographical region. Analyses were performed using descriptive statistics and logistic regression, considering as the outcome variable the use of BZDs. The prevalence found in this study was 7.4% (n = 1094) and was below than others the found in similar studies. It is understood that the most preventive focus of the FHPs may contribute to these findings or the quality of the records have interfered in these results. Users of BZDs studied were in majority in the age range from 56 to 74 years (48.4%), studied only up to the first degree (65.6%), self- mentioned at least one disease (82.2%) and almost half used at least one other psychotropic drug (48.4%) at the same time. There was a significant association between using BZDs and have low educational level (p = 0.005), present self- mentioned diseases (p = 0.000) and the use of other psychotropic drugs (p = 0.000). The association between higher ages and use of BZDs (p = 0.005) was also significant. Analyzes also showed that women who used other psychotropic drugs were five times more likely to use BZDs than those who did not. Women with self- reported disease were almost five times more likely to use BZDs than women who did not report diseases. We concluded that the groups that should receive more attention in relation to the consumption of these substances are those of middle-aged women or older with low schooling, and with chronic disease. It is recommended that the health teams pay attention to the psychosocial needs of persons with mental disorder, discuss and take efforts to implement actions for insertion and social reintegration as well as the expansion of networks of support that can be a strategic aid to the containment of symptoms and improving of the quality of life of these people, in an attempt to avoid the use of more psychotropic drugs and/or of the services of greater complexity

Benzodiazepínicos (BDZ) são fármacos ansiolíticos que se caracterizam por atuar em receptores GABAa presentes no sistema nervoso central. Além dos receptores centrais os BDZ também possuem afinidade por sítios ligantes periféricos presentes em vários tecidos, dentre eles nas glândulas adrenais e em células polimorfonucleares. O presente experimento analisou os efeitos de BDZ sobre a atividade de neutrófilos e sobre os níveis séricos de corticosterona de ratos. Especificamente, o burst e a fagocitose de neutrófilos foram estudados após o tratamento ex vivo e in vitro com diazepam, RO 5-4864, clonazepam e/ou PK 11195. Os efeitos do diazepam sobre a atividade de neutrófilos também foram avaliados após o uso prolongado deste fármaco. Finalmente, os efeitos in vitro dos BDZ foram estudados após a incubação com um bloqueador do canal de cálcio (Ca2+) o L-verapamil. Os resultados mostraram que (1) o tratamento agudo ex vivo com diazepam (10 mg/kg) produziu um aumento do burst oxidativo e da fagocitose induzida por PMA, LPS e Staphylococcus aureus; (2) o tratamento prolongado (21 dias) com diazepam (10 mg/kg) produziu um aumento do burst oxidativo induzido pelos mesmos estímulos e reduziu a fagocitose dos neutrófilos (porcentagem e intensidade); (3) o tratamento agudo ex vivo com diazepam, mas não o prolongado, aumentou os níveis séricos de corticosterona; (4) os efeitos da adição in vitro de diazepam (100 nM) foram na mesma direção daqueles observados após o tratamento agudo; (5) o RO 5-4864 (100 nM) induziu in vitro efeitos similares aos obtidos ex vivo (10 mg/kg) e in vitro (100 nM) com diazepam; (6) o tratamento ex vivo com clonazepam (10 mg/kg) diminuiu o burst oxidativo de neutrófilos induzido por PMA, LPS e Staphylococcus aureus enquanto que o tratamento com este fármaco na dose de 1 mg/kg aumentou o burst oxidativo destas células frente a todos os estímulos; (7) o tratamento ex vivo (1 e 10 mg/kg) e in vitro (100 nM) com clonazepam não alterou a fagocitose de neutrófilos; (8) os efeitos da adição in vitro de clonazepam sobre a atividade de neutrófilos vão na mesma direção daqueles observados após o tratamento ex vivo na dose de 1 mg/kg; (9) o clonazepam (1 e 10 mg/kg) aumentou os níveis séricos de corticosterona; (10) a adição in vitro do PK 11195 (100 nM) per se ou associado com diazepam (100 nM) ou RO 5-4864 (100 nM) aumentou o burst oxidativo e diminuiu a fagocitose realizada pelos neutrófilos; (11) a adição in vitro do PK 11195 (100 nM) associado com o clonazepam (100 nM) diminuiu o burst oxidativo induzido por PMA e diminuiu a fagocitose realizada por estas células; finalmente, (12) a pré incubação por 2 minutos com L-verapamil (100 nM) bloqueou todos os efeitos do diazepam sobre a atividade de neutrófilos estimulada por PMA, LPS e Staphylococcus aureus e reverteu os efeitos do RO 5-4864 no burst oxidativo induzido por PMA. Esses resultados sugeriram que os efeitos dos BDZ não estão relacionados com a ativação de PBR nas células da glândula adrenal e também que eles não estão relacionados a um aumento dos níveis séricos de corticosterona. Indicaram que os BDZ tenham produzido mudanças na concentração do Ca2+ intracelular dos neutrófilos através da ativação de PBR seguida de um aumento da permeabilidade da membrana plasmática ao Ca2+ através da abertura de canais sensíveis ao L-verapamil, permitindo a entrada do Ca2+. Os diferentes efeitos dos BDZ sobre o burst oxidativo e a fagocitose de neutrófilos foram atribuídos a ações específicas deste fármaco em subtipos de PBR presentes nos neutrófilos e nas membranas mitocondriais. Finalmente, os resultados sugeriram o desenvolvimento de tolerância aos efeitos do diazepam sobre os níveis séricos de corticosterona mas não sobre a atividade de neutrófilos. / Benzodiazepines (BDZ) exert their anxiolytic effects via specific binding sites coupled to neuronal GABAa receptors. They also affect a second, pharmacologically different type of receptor, which has been found in many sites such as blood polimorphonuclear cells and adrenals. The present experiment was designed to analyze the effects of BDZ on neutrophil activity and corticosterone serum levels in rats. Specifically, neutrophil oxidative burst and phagocytosis were studied after ex vivo and in vitro treatments with diazepam, Ro 5-4864, clonazepam and/or PK 11195. Diazepam effects on neutrophil activity were also taken after long-term treatment. Finally, in vitro effects of BDZ were studied after incubation with L-verapamil, a Ca2+ channel blocking agent. Results showed that (1) acute and ex vivo diazepam (10.0 mg/kg) treatment induced an increment on PMA, LPS and Staphylococcus aureus induced oxidative burst and phagocytosis (2) long-term (21 days, 10 mg/kg/day) diazepam treatment increased the oxidative burst induced by the same stimuli and reduced both intensity of phagocytosis and the percentage of neutrophil performing phagocytosis; (3) acute and ex vivo diazepam treatment, but not long-term diazepam treatment increased corticosterone serum levels; (4) the effects induced by in vitro addition of diazepam (100 nM) were in the same direction of those observed after acute treatment; (5) Ro 5-4864 (100 nM) induced similar effects to those of diazepam after ex vivo (10.0 mg/kg) and in vitro (100 nM) treatments; (6) ex vivo (10.0 mg/kg) clonazepam treatment decreased the neutrophil oxidative burst induced by PMA and S. aureus after a 1.0 mg/kg dose and decreased these parameters after a higher (10 mg/kg) dose; (7) ex vivo (1.0 and 10.0 mg/kg) and in vitro (100 nM) clonazepam treatments did not change neutrophil phagocytosis; (8) in vitro clonazepam (100 nM) induced effects on neutrophil activity that were in the same direction of those observed after the 1.0 mg/kg dose (9) clonazepam (1.0 and 10.0 mg/kg) increased corticosterone serum levels; (10) in vitro PK 11195 (100 nM) per se or associated with diazepam (100 nM) or Ro 5-4864 (100 nM) increased PMA, LPS and S. aureus induced neutrophil oxidative burst and decreased cell phagocytosis; (11) in vitro PK 11195 (100 nM) together with clonazepam (100 nM) decreased PMA-induced oxidative burst and decreased S. aureus phagocytosis; finally, (12) in vitro L-verapamil (100 nM) incubation for 2 minutes prevented all effects induced by diazepam on PMA, LPS and S. aureus -induced neutrophil activity and reverted the effects induced by Ro 5-4864 (100 nM) on PMA-induced oxidative burst. These results suggest that BDZ effects on neutrophil activity were not related to PBR activation within the adrenal gland cells, leading to increments on corticosterone serum levels. They suggest instead that BDZ induced [Ca2+]i changes in neutrophils through PBR activation followed by an increase in plasma membrane permeability due to L-verapamil sensitive Ca2+ channels and subsequent extracelular Ca2+ entry. The differences of BDZ effects on neutrophil oxidative burst and phagocytosis were attributed to specific actions on different PBR subtypes present on neutrophil and mitochondrial membranes. Finally, results suggest the development of tolerance to diazepam effects on corticosterone serum levels but not on neutrophil activity.