O presente trabalho teve por objetivo estudar a herança genética, determinar o melhor protocolo de extração de DNA para esta espécie, e identificar marcadores moleculares associados à resistência de leiteiro (Euphorbia heterophylla L.) aos herbicidas inibidores da ALS e da PROTOX. A herança genética da resistência foi determinada a partir de cruzamentos entre os biótipos de E. heterophylla suscetível (S) e resistente (R), retrocruzamentos e avanço de geração para F2. A dominância completa da resistência foi comprovada com curvas de dose resposta. Foram testados dez protocolos de extrações de DNA adaptados de métodos descritos na literatura. Os iniciadores específicos para os genes ALS e PROTOX foram desenhados a partir da sequência de DNA consenso destes genes, obtida pelo alinhamento das espécies Manihot esculenta e Ricinus communis. Adicionalmente, foi testada a transferibilidade de vinte marcadores SSRs (sequências simples repetidas) desenhados para o genoma de Manihot esculenta, pois dentre espécies de Euphorbiaceae com maior número de marcadores SSRs desenvolvidos, é a espécie filogeneticamente mais próxima de E. heterophylla. Em relação à herança genética, as frequências observadas nas gerações F1, F2, RCs e RCr não diferiram estatisticamente das frequências esperadas para característica controlada por dois genes dominantes para resistência múltipla e um gene dominante para resistência simples aos inibidores da ALS e PROTOX. Os níveis similares de resistência observados para o heterozigoto F1 e o biótipo homozigoto R, para doses de até 2000 g i.a. ha-1 de fomesafen e doses de até 800 g i.a. ha-1 de imazethapyr, confirmam a dominância completa da resistência aos inibidores da PROTOX e ALS, respectivamente. O protocolo 0,2%BME possibilitou a extração de 7,083 ng μL-1 de DNA, sendo estatisticamente (P=0,05) superior aos demais protocolos. Os compostos fenólicos contaminaram o DNA extraído pelos protocolos FENOL e 3%BME+TB, mas a adição de polivinilpirrolidona (PVP40) no tampão de extração do protocolo 3%BME+TA solucionou este problema. Os iniciadores desenhados para os genes ALS e PROTOX não amplificaram ou não apresentaram polimorfismo visível em gel de agarose entre os biótipos S e R de E. heterophylla. Dez marcadores SSR foram transferidos para E. heterophylla e destes, seis iniciadores apresentaram polimorfismo entre os biótipos S e R. / This study aimed to assess the genetic inheritance, determine the better DNA isolation protocol for this species and to identify molecular markers associated with the Wild Poinsettia (Euphorbia heterophylla L.) resistance ALS- and PROTOX- inhibiting herbicides and. The genetic inheritance of resistance was determined from crosses between E. heterophylla biotypes susceptible (S) and resistant (R), backcrosses and F2 generation. The complete dominance of resistance was confirmed with dose response curves. Ten adjusted methods for DNA isolation described in the literature were tested. The specific primers for ALS and PROTOX genes were designed from the consensus DNA sequence of these genes, obtained by aligning the gene sequences of the species Manihot esculenta and Ricinus communis L. Additionally, it was assessed the transferability of twenty SSR (simple sequence repeat) markers designed for Manihot esculenta, because among the species of Euphorbiaceae with more developed SSRs markers, because it is the closest relative phylogenetic species of E. heterophylla. Regarding genetic inheritance, the frequencies observed in the F1, F2, RCs and RCr did not differ significantly from the expected frequencies for a trait controlled by two dominant genes for multiple resistance and a single dominant gene for simple resistance to ALS- and PROTOX-inhibiting herbicides. The similar levels of resistance to dosage up to 2000 g i.a. ha-1 of fomesafen and dosage up to 800 g i.a. ha-1 of imazethapyr observed in F1 (heterozygous) and homozygous R biotype confirm the complete dominance of resistance to PROTOX- and ALS-inhibiting herbicides, respectively. The 0.2%BME protocol allowed the isolation of 7,083 ng μL-1 DNA, significantly (P=0.05) higher than other methods. Co-isolation of phenolic compounds was observed in FENOL and 3%BME+TB methods, but the addition of polyvinylpyrrolidone (PVP40) in the protocol extraction buffer 3%BME+TA solved this problem. The primers designed for ALS and PROTOX genes amplified but not showed no visible polymorphism in agarose gel between the S and R biotypes of E. heterophylla. Regarding the SSR transferability, ten markers were transferred to E. heterophylla, however, these six primers showed polymorphism among S and R biotypes.

Made available in DSpace on 2018-06-27T18:56:35Z (GMT). No. of bitstreams: 2 Luciana dos Santos Lopes.pdf: 903442 bytes, checksum: 9241c2c46a0d205e6d9e1b049687e5f7 (MD5) Luciana dos Santos Lopes.pdf.jpg: 3030 bytes, checksum: 0c80e97bb571acad00dbccdd7ce7c921 (MD5) Previous issue date: 2009-01-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The NTPDase (apirase-ect, ect-difosfoidrolase, CD39, EC 3.6.1.5) hydrolyses tri nucleotides and/or diphosphate. It is an ectonucleotidase, identified as the antigen surface of lymphoid cells, whose greatest expression leads to an increase in activity of ATPase and ADPase and these cells through the hydrolysis of ATP and ADP. The inhibitor of ecto-nucleotidases may represent valuable therapeutic tools to amplify the biological effects of extracellular nucleotides, since they induce apoptosis in lymphocytes, causing a state of immunosuppression. It is known through the literature that sodium azide produces a significant inhibition in the hydrolysis of ATP and ADP, caused this enzyme when it is used in concentrations around 20mM. The azides are chemical components widely used in organic synthesis, and remarkably stable in biological environment. The aim of this study was to determine the inhibitory effects of an organic azide derived from acetic acid in the hydrolysis of ATP and ADP, which is the activity of NTPDase of human lymphocytes. Since changes in distribution and activity of this enzyme have been reported in various pathological conditions, demonstrating its participation in the activation of lymphocyte, it is believed that the identification of new compounds inhibiting NTPDase will contribute to treatment of diseases where there is a need for immunosuppression, as allergies and autoimmune diseases. After incubation of lymphocytes in medium containing different concentrations of azide organic, there was a significant inhibition of ADP in the hydrolysis when used concentrations of 10 and 20 mM, and this of 50 and 77% respectively. Unverified inhibitory effects on the ATP hydrolysis with organic azide. A changed the hydrolysis of nucleotide adenine diphosphate, indicating that the substance could be used as an inhibitor of mixed NTPDase. / A NTPDase (ecto-apirase, ecto-difosfoidrolase, CD39, EC 3.6.1.5) hidrolisa nucleotídeos tri e/ou difosfatados. É uma ectonucleotidase, identificada como antígeno de superfície de células linfóides, cuja maior expressão leva a um aumento das atividades de ATPase e ADPase nestas células. Os inibidores de ecto-nucleotidases podem representar valiosas ferramentas terapêuticas para amplificar os efeitos biológicos dos nucleotídeos liberados no meio extracelular, uma vez que estes induzem apoptose nos linfócitos, causando um estado de imunodepressão. Sabe-se através da literatura que azida sódica, quando utilizada em concentrações em torno de 20mM, produz uma significativa inibição na hidrólise de ATP e ADP causada por esta enzima. As azidas são componentes químicos muito usados em sínteses orgânicas, sendo notavelmente estáveis em ambiente biológico. Este trabalho, procurou determinar os efeitos inibitórios de uma azida orgânica derivada do ácido acético na hidrólise de ATP e ADP, ou seja, na atividade da NTPDase de linfócitos humanos. Uma vez que alterações na atividade e distribuição desta enzima têm sido relatadas em várias condições patológicas, demonstrando sua participação na ativação do linfócito, acredita-se que a identificação de novos compostos inibidores da enzima NTPDase venha a contribuir em patologias onde há necessidade de uma imunossupressão, como em alergias e doenças autoimunes. Após incubação dos linfócitos em meio contendo diferentes concentrações de azida orgânica, verificou-se uma inibição significativa na hidrólise do ADP quando utilizadas as concentrações de 10 e 20 mM, sendo esta de 50 e 77%, respectivamente. Não foram verificados efeitos inibitórios sobre a hidrólise do ATP. A azida orgânica alterou a hidrólise dos nucleotídeos da adenina difosfatados, indicando que esta substância poderia ser utilizada como um inibidor da NTPDase, cujo tipo de inibição causada seria mista.

O estabelecimento de uma camada híbrida adequada no canal radicular representa um dos principais desafios clínicos devido à dificuldade de acesso. Dessa forma, o uso de inibidores proteolíticos poderia tornar-se um recurso favorável. O objetivo deste estudo foi avaliar o efeito de inibidores proteolíticos na união de pino de fibra de vidro fixado com cimento adesivo, considerando os terços radiculares e tempos distintos, por meio da resistência de união (RU). Cento e quarenta e quatro raízes bovinas foram selecionadas e divididas em 6 grupos de tratamento, e redivididas em 3 subgrupos de acordo com os tempos de avaliação de 24 horas, 6 e 12 meses (n=8). Após o tratamento endodôntico e desobturação padronizados, as raízes foram cimentadas com pinos de fibra de vidro cônicos (Exacto/Angelus). As raízes foram tratadas com sistema adesivo convencional de três passos, Adper Scotchbond Multipurpose/ 3M ESPE (SBMP) e cimento dual RelyX ARC/ 3M ESPE. Após prévia divisão, foram alocadas em grupos CN (Controle Negativo- sem pré tratamento associado), CP (Controle Positivo- com agentes ativador e catalisador), EDTA (ácido etilenodiamino tetra-acético a 17%), CHX (digluconato de clorexidina a 2%), E-5 (E- 64 a 5 M) e E-10 (E-64 a 10 M). Após 24 horas, as raízes foram seccionadas perpendicularmente ao longo eixo e identificadas quanto à região, obtendo-se fatias de 1 mm de espessura (cervical, médio e apical), que foram armazenadas em saliva artificial para serem testadas. Todas as fatias foram submetidas ao teste de extrusão (push-out) na máquina de teste universal (Instron) com célula de carga de 50 N a 0,5 mm/min. Os dados foram analisados pelo teste de ANOVA a três critérios e comparações múltiplas com Tukey, ambos com p<0,05. Após 24 horas, não se observou diferenças entre os tratamentos. Após 6 meses, a CHX demonstrou melhor desempenho, cujo efeito não se prorrogou até os 12 meses. O uso de inibidores proteolíticos não foram capazes de preservar a resistência de união dos pinos intrarradiculares até o tempo de 12 meses. / The adequate establishment of hybrid layer in the root canal on bonding process is still a clinical challenge due to its hard access. Thus, the use of proteolytic inhibitors could become a favorable tool. The aim of this study was to evaluate the effect of proteolytic inhibitors in the bonding of a glass- fiber post fixed with a luting cement, regarding the root thirds and different times through the bond strength. One hundred and forty four bovine roots were selected and divided into 6 treatment groups, and subdivided according to the time of evaluation of 24 hours, 6 and 12 months (n=8). After endodontic treatment and standardized removal procedure, the roots were cemented with tapered glass fiber posts (Exacto/ Angelus). The roots were treated with three-step etch-and-rinse adhesive system Adper Scotchbond Multipurpose/ 3M ESPE (SBMP) and dual cement RelyX ARC/ 3M ESPE. After previous division, CN (negative- control without pre associated treatment), CP (Control positive- with activator and catalyst agents) EDTA (17% ethylenediaminetetraacetic acid) CHX (2% chlorhexidine digluconate) E-5 (5M E-64) and E-10 (10M E-64). After 24h, the roots were sectioned perpendicular to the long axis and identified according to third in 1mm thick slices (cervical, middle and apical), which were stored in artificial saliva to be tested. All slices were subjected to extrusion tests (push-out) in the universal test machine (Instron) at 50 N load cell at 0.5 mm/min. Data were analyzed with three-way ANOVA and multiple comparisons with Tukey test, both with p <0.05. After 24 hours, no differences were observed between treatments. After 6 months, CHX showed better performance, which did not last up 12 months. The proteolytic inhibitors performed differently in the bonding process over time; only CHX promoted inhibition at 6 months. The use of proteolytic inhibitors were not able to maintain the bond strength of intraradicular posts up time of 12 months.

Orientador: Anita Jocelyne Marsaioli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-25T07:45:46Z (GMT). No. of bitstreams: 1 Martins_LucasGelain_D.pdf: 5948717 bytes, checksum: d64f37183bfefec1cedc88b263d32340 (MD5) Previous issue date: 2014 / Resumo: Nesta tese consiste no estudo de interações supramoleculares utilizando diferentes metodologias de Ressonância Magnética Nuclear de hidrogênio (RMN de 1H), tais como ROESY 1D, RMN-DOSY e RMN-STD. Os sistemas supramoleculares foram abordados em dois casos de estudo diferentes. O Capítulo I tem como objeto de estudo interações do fármaco Dapsona com diferentes carreadores de fármacos (ß-CD, SBE-ß-CD e lipossoma de EPC), com a finalidade de encontrar formulações nas quais a Dapsona seja mais solúvel. Os complexos binários e ternário formados foram determinadas por medidas de difusão molecular. Foram observadas as formações dos complexos Dap/ß-CD, Da-/SBE-ß-CD, Dap/EPC e o ternário Dap/ß-CD/EPC, os quais contribuem para o aumento de solubilidade do fármaco. O objeto de estudo apresentado no Capítulo II é a inibição da enzima acetilcolinesterase por dois alcaloides, a Fisostigmina e a Crinina. Para determinação das constantes de dissociação aparentes foram utilizados os crescimentos iniciais das curvas de saturação obtids por RMN-STD para construção das Isotermas de Langmuir. De acordo com os valores de constantes obtidos foi possível concluir que a AchE tem mais afinidade com a Fisostigmina do que com a Crinina / Abstract: This thesis consists of the supramolecular interactions study applying different Nuclear Magnetic Resonance methodologies (1H NMR) such as 1D ROESY, DOSY-NMR and STD-NMR. The supramolecular systems were addressed in two different case studies. In chapter I Dapsone solubility was the case study. To solve the solubility problem the interactions between Dapsone and various drug carriers (ß-CD , SBE-ß-CD and EPC liposome) were characterized in terms of the binary and ternary complexes structure using NMR method, such as 1D ROESY and STD-NMR and apparent association constants were determined by measuring molecular diffusion using DOSY-NMR. Dap/ß-CD, Dap/SBE-ß-CD, Dap/EPC and Dap/ß-CD/EPC complexes were observed and Dapsone water solubility was increased. The case study in Chapter II is the inhibition of acetylcholinesterase by two alkaloids, Physostigmine and Crinina. The apparent dissociation constants were determined using the initial growth saturation curves obtained by STD-NMRto construct the Langmuir isotherms. The constants showed that the AChE has more affinity for the Physostigmine than Crinine / Doutorado / Quimica Organica / Doutor em Quimica

Abstract HIF prolyl 4-hydroxylases (HIF-P4Hs) and HIF asparaginyl hydroxylase (FIH) are novel members of the 2-oxoglutarate dioxygenase family that play key roles in the regulation of the hypoxia-inducible transcription factor (HIF). They hydroxylate specific proline and asparagine residues in HIF-α, leading to its proteasomal degradation and inhibition of its transcriptional activity, respectively. These enzymes are inhibited in hypoxia, and as a consequence HIF-α becomes stabilized, forms a dimer with HIF-β, attains its maximal transcriptional activity and induces expression of many genes that are important for cell survival under hypoxic conditions. The three HIF-P4Hs and FIH were expressed here as recombinant proteins in insect cells and purified to near homogeneity. All these enzymes were found to require long peptide substrates. The three HIF-P4Hs and FIH acted differently on the various potential hydroxylation sites in the HIF-α isoforms. The HIF-P4Hs acted well on sequences with cores distinctly different from the core motif -Leu-X-X-Leu-Ala-Pro-, suggested based on sequence analysis studies, the alanine being the only relatively strict requirement in addition to the proline itself. Acidic residues around the hydroxylation site also played a distinct role. These results together with those of others provide evidence that there is no conserved core motif for the hydroxylation by HIF-P4Hs. The Km values of the HIF-P4Hs for O2 were slightly above its atmospheric concentration, while the Km of FIH was about one-third of these values but still 2.5 times that of the type I collagen P4H. The HIF-P4Hs are thus effective oxygen sensors, as even small decreases in the amount of O2 affect their activities, while a more severe decrease is required to inhibit FIH activity. Small molecule inhibitors of the collagen P4Hs also inhibited the HIF-P4Hs and FIH but with distinctly different Ki values, indicating that it should be possible to develop specific inhibitors for the HIF-P4Hs and FIH. The HIF-P4Hs were found to bind the iron cosubstrate more tightly than FIH and the collagen P4Hs, and the chelator desferrioxamine was an ineffective inhibitor of the HIF-P4Hs in vitro. Several metals were effective competitive inhibitors of FIH but they were ineffective inhibitors of the HIF-P4Hs. The well-known stabilization of HIF-1α by cobalt and nickel is thus not due to a simple competitive inhibition of the HIF-P4Hs, and is probably at least in part due to HIF-P4H-independent mechanisms. The effective inhibition of FIH by these metals nevertheless indicates that the stabilized HIF-1α is transcriptionally fully active.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Cardiovascular disease is a major health problem and remains the number one cause of death worldwide. For centuries, medicinal plants have been used in different cultures as medicines for the treatment and control of various diseases. Olea africana, also known as the wild olive, is amongst the herbal plants used by people to treat many ailments.Recently, scientific studies on the hypotensive, vasodilatory and antidysarrhythmic effects of O. africana have been reported. Triterpenoids isolated from the O. africana leaves, for example, have antioxidant properties. The aqueous extract from the leaves of O. africana also have angiotensin-converting enzyme (ACE) inhibitory effects. ACE inhibitors and antioxidants protect the heart against ischemic-reperfusion injury. The serine / threonine protein kinase B (PKB) also known as Akt is activated downstream of phosphoinositide 3- (PI-3) kinase (PI-3-Kinase) and is involved in cardioprotection against ischemia-reperfusion injury. Angiotensin II (AII) decreases the intrinsic PI-3-kinase activity. In this study, we hypothesized that ACE inhibitors increase PI-3-kinase activity and thus activates PKB. The aims of this study were: 1) to determine whether treatment with the crude aqueous extract of leaves of O. africana protect the heart against ischemic-reperfusion injury and 2) if so, to determine whether the protection is mediated via the PKB signaling mechanism. Hearts isolated from male Wistar rats were perfused with different concentrations of the plant extract. In one set of experiments, male Wistar rats were treated with the plant extract (1000 mg/kg/day) for 5 weeks for the evaluation of cardiac function before and after ischemia. At the end of the experiments, hearts were freeze-clamped and kept for PKB / Akt determination. In another set of experiments, we determined the effect of O. africana extract (1000 mg/kg/day) or captopril (50 mg/kg/day) on infarct size. Rats fed jelly served as controls for captopril. In a subset of experiments, hearts were frozen immediately after treatment with O. africana extract (1000 mg/kg/day) or captopril (50mg/kg/day) and PKB were determined.Perfusion with the plant extract significantly decreased coronary flow (p<0.05). The heart function was decreased as evidenced by observed decreases in the force of contraction and heart rate, although these were not measured. Chronic treatment with the crude aqueous plant extract had no effect on cardiac function before ischemia, functional recovery (% left ventricular developed pressure and % rate pressure product) and PKB /Akt phosphorylation (p>0.05). Both the aqueous extract of O. africana leaves and captopril had no effect on infarct size compared to the control group (p>0.05). Captopril,however, improved the recovery of the left ventricular developed pressure. Non-perfused hearts isolated from rats treated with O. africana extract and captopril did not show any response to both captopril and the O. africana extract treatment as measured by PKB /Akt phosphorylation. The results of the present study suggest that the crude aqueous extract of O. africana is not cardioprotective against ischemia-reperfusion injury in this system of the isolated perfused rat heart.

Criptococose é uma doença causada pelo fungo C. neoformans que têm grande importância atualmente, devido ao aumento da população imunocomprometida,. Além disso, existem poucas opções terapêuticas contra micoses profundas. Neste trabalho foi avaliado se a via de biossíntese do triptofano seria um bom alvo para o desenvolvimento de novos antifúngicos. Com o uso da tecnologia de RNA de interferência, concluiu-se que esta via de síntese é essencial para a sobrevivência desta levedura, sendo, portanto, um ótimo alvo. Ainda neste estudo, demonstrou-se que a letalidade decorre da baixa captação de triptofano pelas permeases de aminoácidos, as quais sofrem repressão catabólica pela fonte de nitrogênio e efeito negativo da temperatura. Foram testados dois inibidores específicos que atuam sobre a antranilato sintase e a triptofano sintase, duas enzimas cruciais para a conversão do corismato em triptofano. Ambos compostos causaram inibição do crescimento de C. neoformans e C. gattii. / Cryptococcosis is a disease caused by C. neoformans, currently of great importance due to the increase in immunocompromised population. Furthermore, there are few therapeutic options for treating this disease. This study evaluated the tryptophan biosynthetic pathway as a possible target for the antifungal development. By using RNA interference technology we concluded that this metabolic pathway is essential for the survival of this yeast, and, therefore, it is a good target. In the same study, it was demonstrated that lethality results from the low uptake of the tryptophan amino acid by permeases, which undergo nitrogen catabolite repression and negative effect of temperature. Two specific inhibitors acting on the anthranilate synthase and tryptophan synthase, two key enzymes for the conversion of chorismate into tryptophan were tested. Both compounds caused growth inhibition of C. neoformans and C. gattii.

In this review we discuss the role of ATP synthase as a molecular drug target for natural and synthetic antimicrobial/ antitumor peptides. We start with an introduction of the universal nature of the ATP synthase enzyme and its role as a biological nanomotor. Significant structural features required for catalytic activity and motor functions of ATP synthase are described. Relevant details regarding the presence of ATP synthase on the surface of several animal cell types, where it is associated with multiple cellular processes making it a potential drug target with respect to antimicrobial peptides and other inhibitors such as dietary polyphenols, is also reviewed. ATP synthase is known to have about twelve discrete inhibitor binding sites including peptides and other inhibitors located at the interface of α/β subunits on the F1 sector of the enzyme. Molecular interaction of peptides at the β DEELSEED site on ATP synthase is discussed with specific examples. An inhibitory effect of other natural/synthetic inhibitors on ATP is highlighted to explore the therapeutic roles played by peptides and other inhibitors. Lastly, the effect of peptides on the inhibition of the Escherichia coli model system through their action on ATP synthase is presented.

In this review we discuss the role of ATP synthase as a molecular drug target for natural and synthetic antimicrobial/ antitumor peptides. We start with an introduction of the universal nature of the ATP synthase enzyme and its role as a biological nanomotor. Significant structural features required for catalytic activity and motor functions of ATP synthase are described. Relevant details regarding the presence of ATP synthase on the surface of several animal cell types, where it is associated with multiple cellular processes making it a potential drug target with respect to antimicrobial peptides and other inhibitors such as dietary polyphenols, is also reviewed. ATP synthase is known to have about twelve discrete inhibitor binding sites including peptides and other inhibitors located at the interface of α/β subunits on the F1 sector of the enzyme. Molecular interaction of peptides at the β DEELSEED site on ATP synthase is discussed with specific examples. An inhibitory effect of other natural/synthetic inhibitors on ATP is highlighted to explore the therapeutic roles played by peptides and other inhibitors. Lastly, the effect of peptides on the inhibition of the Escherichia coli model system through their action on ATP synthase is presented.

Background: Amiodarone (AM) is a widely used anti-arrhythmic medication. Its utility is, however, limited by adverse side effects. The mechanism of amiodarone-induced toxicity (APT) in the lungs is attributed primarily to stimulation of the angiotensin enzyme system leading to lung cell apoptosis and cell death. This mechanism has been demonstrated by in vitro and in vivo experimental animal studies. To date, however, no in vivo human studies have confirmed this mechanism for APT. Purpose: This study was undertaken to determine whether angiotensin converting enzyme inhibitors (ACE-I) or angiotensin receptor blockers (ARB) offer a protective effect against APT in humans. Demonstration of a protective effect of an ACE-I or ARB would suggest that stimulation of the angiotensin enzyme system may be a key process in APT. Design: An 8-year retrospective analysis of all patients on AM therapy at the James H. Quillen Veterans Affairs Medical Center was undertaken. Results: A total of 1000 patients on AM were identified. One-hundred-and-seventeen were excluded from the study. Five-hundred-and-twenty-four patients were simultaneously on an ACE-I or ARB. The remaining 359 patients were not. Pulmonary toxicity attributed to AM was identified in five and 14 patients with and without concomitant ACE-I or ARB therapy, respectively. The APT rate for the entire patient sample was 2.2%. APT occurred in 1% of patients on an ACE-I or ARB and in 3.9% of patients not taking an ACE-I or ARB. This observed difference in percentage of APT was statistically significant. Conclusion: The concomitant use of ACE-I or ARB in patients taking AM appears to offer a protective effect against APT. This observation suggests that the stimulation of the angiotensin enzyme system may play an important role in APT in humans.