A research report submitted to the Faculty of Health Sciences, University of the Witwatersrand, in partial fulfilment of the requirements for the degree of Master in Science ( Medicine ) Pharmaceutical Affairs / Lippia javanica is a widely spread woody shrub and the major traditional use is reflected in its vernacular name; fever tea ' koorsbossie '. An infusion of the leaves is also used as a decongestant for colds and coughs. Infusions may also be used topically to treat scabies and lice A preliminary study indicated that the essential oil chemistry varies dramatically both within and between natural plant populations. / IT2018

ABSTRACT The formulation and approval of the Operational Plan for Comprehensive Care, Management and Treatment for HIV/AIDS in 2003 was a major victory for the roll-out of anti-retroviral therapy (ART) in the public sector in South Africa. Since its initiation in 2004, the ART Programme has expanded rapidly and realised considerable gains in prolonging life. However, it has also faced major constraints and implementation has been uneven across provinces. This study investigates the impact of organisational capacity upon levels of adherence to ART in two public sector sites in Gauteng. The study uses the Chronic Care Model (CCM) proposed by Edward Wagner (2004). The CCM identifies four major components as crucial to effective clinical outcomes for the management of chronic care. These factors are (1) prepared proactive practice teams; (2) delivery systems design; (3) decision support; and (4) clinical information systems. Both sites demonstrated different strengths and constraints. Strengths included the presence of motivated champions leading the ART service, positive patient-provider relationships, shifting of tasks to lower level health workers to deal with the shortage of skilled staff, good relationships with non-governmental organisations and the innovation to deal with challenges in a way that does not compromise the quality of care provided to patients using the CCMT service. Overall constraints that were identified in the two facilities include the shortage of skilled staff, burn-out among staff, a shortage of space, inconsistent data collection and interpretation, as well as poor integration and collaboration between local and provincial government in relation to the shared responsibility for the provision of Tuberculosis (TB) treatment and other related CCMT services. Despite these barriers, levels of adherence exceeded 85% in both sites.

Ph.D., Faculty of Health Sciences, University of the Witwatersrand, 2011 / The HIV-­‐1 pandemic continues unabated. Although treatments exist that can substantially alleviate the morbidity and mortality associated with HIV, there is still a need for improved anti-­‐HIV treatments that reduce toxicities and administration frequency and mediate sustained inhibition of viral replication. Given the high mutability and variability of the virus, a strategy that is garnering increasing focus is the targeting of host factors that the virus requires to replicate, so-­‐called HIV-­‐dependency factors (HDFs). It is hoped this will reduce the emergence of viral drug resistance. A number of genome-­‐wide screens have been performed to identify HDFs, although many remain to be validated, particularly in relevant cells lines. An objective of this thesis was to validate three host factors as HDFs, in both TZM-­‐bl reporter and T cell-­‐derived cell lines, and to examine their potential as anti-­‐HIV-­‐1 therapeutic targets through exploitation of the cellular gene silencing pathway, RNA interference (RNAi). These were HIV-­‐1 Tat specific factor 1 (HTATSF1), DEAD (Asp-­‐Glu-­‐Ala-­‐Asp) box polypeptide 3, X-­‐ linked (DDX3X) and SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily b, member 1 (SMARCB1), selected because they had been previously implicated in HIV-­‐ 1 pathogenesis. The well-­‐characterised HDF, PC4 and SFRS1 interacting protein 1 (PSIP1)/lens epithelium-­‐derived growth factor (LEDGF)/p75, was included in the study as a positive control. Cassettes expressing short hairpin RNAs (shRNAs) targeting the four host proteins were generated, although shRNAs did not suppress endogenous ddx3x mRNA levels. The ability of shRNAs to inhibit HIV-­‐1 replication in the reporter cell line, TZM-­‐bl, was examined. These HeLa-­‐ derived cells are permissive for R5-­‐tropic HIV-­‐1 infection and contain an integrated luciferase gene driven by the viral promoter. shRNAs mediated a dose-­‐dependent inhibition of luciferase activity in cells infected with a HIV-­‐1 subtype B molecular clone and, although production of the viral protein p24 was unaltered, infectious particle production was decreased in cells treated with a shRNA suppressing HTATSF1. Little effect was observed with a shRNA targeting SMARCB1, suggesting that this may not function as an HDF under these conditions. No effect on infectious particle production was seen with the shRNA targeting PSIP1, which was a result of the long half-­‐ life of this protein, highlighting a limitation of using such reporter systems for HDF validation. Importantly, shRNAs were not associated with any cytotoxic effects in TZM-­‐bl cells. Whether HTATSF1 is a potential therapeutic target was interrogated further in the more relevant T cell-­‐derived SupT1 cell line. Lentiviruses were used to generate populations where >90% had one copy of the integrated shRNA expression cassette. Replication of the subtype B molecular clone p81A-­‐4 was significantly inhibited in the shH1-­‐expressing SupT1 cell line, which targets HTATSF1, for over 14 days post-­‐infection, although inhibition was not as pronounced asthat observed in the shP1-­‐expressing SupT1 cell line, which targets PSIP1. In contrast to a previous report, no change in the ratio of unspliced to singly-­‐ or multiply-­‐spliced HIV-­‐1 transcripts were detected in shH1-­‐expressing SupT1 cells, suggesting that HTATSF1 does not function as a splicing cofactor in this system. A slight rebound in p24 levels at 14 days post-­‐infection was accompanied by increased HTATSF1 expression and a decrease in the percentage of cells with transgene expression in the population. In addition, there was a slight decrease in shH1-­‐derived guide strand expression, but no change in transcription rates of the htatsf1 gene, suggesting that cells within the population with shH1 expression and HTATSF1 suppression may have a growth disadvantage. Thus, although this work demonstrates for the first time that HTATSF1 functions as an HDF in T cell-­‐derived SupT1 cells, it may not constitute a viable therapeutic target. A second objective of this thesis was to examine the feasibility of transcriptional gene silencing (TGS) of HDFs as an anti-­‐HIV strategy. TGS is a small RNA-­‐induced gene silencing pathway that operates through chromatin remodelling with the potential to mediate long-­‐term silencing of gene expression. Thus, its application may reduce the frequency of drug administration and associated toxicities. Short interfering RNAs (siRNAs) targeting the htatsf1 promoter were able to reduce target mRNA expression, which was accompanied by decreased htatsf1 transcription rates in HEK293T cells, suggesting silencing via a TGS mechanism. The htatsf1 silencing inhibited infectious HIV-­‐1 particle production from TZM-­‐bl cells. This work provides proof of principle that TGS induction at a HDF may inhibit HIV-­‐1 replication. siRNAs targeting the ddx3x promoter did not induce TGS. To examine whether gene susceptibility to TGS may be influenced by promoter architectures, 49 promoter features were examined for enrichment in genes at which small RNA-­‐induced TGS has been reported. Initially, the TGS group was compared to a random set of 2,000 promoters and then all other promoters in the genome. To control for gene activation, two further analyses were performed comparing the TGS group features to those from promoters active in the THP-­‐1 cell line and housekeeping genes. Whilst difficult to ascribe differences between the TGS group and the control groups to anything beyond a variation in the proportion of active genes within each group, there was enrichment for certain promoter features that are independent of activity; the TGS group was characterised by broad transcription start regions, high CpG content and a single expression profile. Moreover, the fraction of promoters with reported non-­‐coding RNA overlap was greater in the TGS group than the control groups. Thus, there is some evidence that a number of promoter features are associated with TGS susceptibility. It is hoped this novel analysis will facilitate selection of future TGS targets, including HDFs. In summary, the work presented in this thesis paves the way for development of improved anti-­‐HIV therapies involving HDF-­‐targeted TGS-­‐based gene therapies that mediate sustained inhibition of the virus.

A RESEARCH REPORT SUBMITTED TO THE FACULTY OF HEALTH SCIENCES, UNIVERSITY OF WITWATERSRAND, JOHANNESBURG, IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF PHARMACY IN CLINICAL PHARMACY 2004 / The purpose of the study was to evaluate the use of cefotaxime with the idea of improving its use within the hospital. Improving the use of cefotaxime could result in a change in the proportion spent from the pharmacy budget. More importantly a change in prescribing patterns of this drug could also result in a reduction in resistant patterns of cefotaxime. / IT2018

O presente trabalho propõe comparar os escritos de dois ideólogos pertencentes a dois importantes movimentos políticos do século XX: um do integralismo brasileiro e outro do nacional-socialismo alemão respectivamente: Gustavo Barroso e Gottfried Feder. Após uma leitura atenta das obras de cada um dos autores, redigidas nos anos 1920 e 1930, compreendemos que o conceito de capital financeiro foi de fundamental importância para que ambos desenvolvessem uma narrativa conspiratória que denunciava um complô judaico de conquista do mundo. Valendo-nos da História Conceitual, percebemos que as obras de Barroso e Feder significaram um momento-chave na história do conceito de capital financeiro, especialmente tendo em vista que ambos escreviam à luz da primeira hegemonia financeira. Entretanto, enquanto Barroso defendia uma luta de caráter universal contra o capital financeiro, Feder achava que tal luta deveria ser protagonizada pelo povo alemão. Ademais, se, por um lado, Barroso conseguiu permanecer como uma figura de proa do movimento integralista até a sua extinção em 1937, Feder, por outro, acabou marginalizado no cenário político após a ascensão nazista ao poder em 1933. / This paper aims at comparing the writings of two ideologues who belonged to two important political movements from the 20th century: one from the Brazilian integralism, and the other from German national-socialism Gustavo Barroso and Gottfried Feder, respectively. After a careful reading of these authors works, written on the 1920es and 1930es, we managed to understand that the concept of financial capital was of main importance for both authors to develop a conspiratorial narrative that uncovered a Jewish plot to conquer the world. Based on the Conceptual History, we noticed that the works of Barroso and Feder represented a key moment in the History of the concept of financial capital, especially considering that both of them wrote during the first financial hegemony. However, while Barroso, on the one hand, thought that the struggle against financial capital should be a universal struggle, Feder, on the other, believed that such struggle should have only the German people as its leading actor. Moreover, while Barroso managed to play a major role in the integralist movement until its ending on 1937, Feder ended up marginalized in the Nazi political scenario after the national-socialist seizure of power on 1933.

Desde a entrada dos visigodos nas terras do Império Romano percebemos uma intenção clara da aristocracia dirigente de fixação do povo em um território e de normatização de um poder sistemático. Ao longo dos séculos IV ao VII esse processo esbarrou em diversos fatores, como as disputas entre as aristocracias pelo poder e a fragilidade da transmissão do poder entre os visigodos, que não possuíam o critério hereditário para isso. Dessa forma, a partir do governo de Leovigildo notamos uma tentativa de normatização política e de reforço da autoridade do rei e da monarquia por meio da unidade religiosa. Para conquistar essa unidade religiosa não alcançada por Leovigildo, seu filho Recaredo buscou o apoio legitimador da Igreja Católica. A partir desse episódio, os governantes que o sucederam também deram continuidade a essa política de unificação religiosa, o que contribuía para o fortalecimento do poder real e da monarquia enquanto instituição.Para buscar essa unidade religiosa os cânones conciliares da Península Ibérica passaram a sistematizar um vasto corpo de legislação antijudaica. Nesse sentido, questionamos se essas medidas contribuíam para o reforço da unidade religiosa e política na região, além de contribuir para o reforço da identidade entre a aristocracia católica, já que agora esses possuíam um inimigo em comum para combater, no caso os judeus. Essa união gerada para combater um inimigo compartilhado pode ter favorecido a governabilidade na região, já que o rei é quem liderava esse processo de combate àqueles que comprometiam a salvação do reino. / Since the entry of the Visigoths in the lands of the Roman Empire perceive a clear intention of the ruling aristocracy attachment of the people in a territory and standardization of a systematic power. Over the centuries IV to VII this process ran on several factors, such as disputes between the aristocracy and the fragility of the power transmission of power between the Visigoths, who had no hereditary criterion for this. Thus, from the government Leovigild noticed an attempt to standardize policy and strengthening the authority of the king and the monarchy through religious unity. To conquer this religious unity not achieved by Leovigild his son Reccared sought support legitimizing the Catholic Church. From this episode, the rulers who succeeded him also continued this policy of religious unity, which contributed to the strengthening of royal power and the monarchy as an institution. To get that religious unity conciliar canons of the Iberian Peninsula began to systematize a large body of anti-Jewish legislation. Accordingly, we question whether these measures contributed to strengthening the unity of religion and politics in the region and contribute to the strengthening of the identity of the Catholic aristocracy, now that these had a common enemy to fight, if the Jews. This union created to fight an enemy may have favored the shared governance in the region, as the king who is leading this process to combat those who committed the salvation of the kingdom.

Desde os primórdios da medicina, substâncias químicas derivadas de animais, vegetais e microrganismos têm sido usadas no tratamento de diversas doenças. Dentre estas, os produtos derivados de vegetais têm dominado as farmacopeias por milhares de anos, fornecendo uma fonte inesgotável de recursos medicinais. O metabolismo secundário geralmente é definido como o responsável pela grande variedade de substâncias químicas apresentadas pelas plantas, porém específicos para alguns grupos. Aos flavonoides atribuem-se importantes funções fisiológicas e ecológicas para o vegetal, como proteção contra a radiação e atração de polinizadores. Para os seres humanos, substâncias dessa classe de metabólitos possuem ação medicinal atuando principalmente como agentes anti-inflamatórios, antioxidantes, antitumorais, hepatoprotetores, antimicrobianos e antivirais. O segundo maior gênero de Euphorbiaceae é Croton L., com aproximadamente 1300 espécies arbóreas, arbustivas ou herbáceas, distribuídas em zonas tropicais e subtropicais do Novo e Velho Mundo. Em praticamente todos os ecossistemas brasileiros encontram-se representantes de Croton. Croton cajucara é uma espécie em testes clínicos, tendo indicações terapêuticas para diabetes, distúrbios hepáticos e renais, anti-inflamatório, antimicrobiano, entre outras. Croton sphaerogynus Baill. por emergir no mesmo clado que C. cajucara, torna-se promissora na prospecção de substâncias bioativas. Este trabalho teve como objetivo investigar a composição fenólica e o potencial biológico de substâncias fenólicas presentes nos extratos foliares de C. sphaerogynus por meio de ensaios antioxidantes, antimicrobianos e anti-HIV-1. Como principais resultados, C. sphaerogynus se caracterizou como uma espécie produtora exclusivamente de flavonóis, além de ser grande produtora de diterpenos. Dois flavonoides foram isolados, quercetina 3,7-dimetil éter e campferol 3-metil éter. Quanto às atividades biológicas, a subfração composta majoritariamente por quercetina 3,7-dimetil éter apresentou alto potencial antioxidante e a maior atividade anti-HIV-1 entre os extratos e outras subfrações testadas. A fração hexânica, rica em diterpenos, foi a que apresentou atividade antimicrobiana promissora. Dessa forma, C. spherogynus torna-se uma espécie promissora para o uso de substâncias antioxidantes / Since the early days of medicine, chemical substances derived from animals, plants and micro-organisms have been used in treating various diseases. Among these, the products derived from plants have dominated the pharmacopoeia for thousands of years, providing an inexhaustible source of medicinal resources. Secondary metabolism is generally defined as responsible for the wide variety of chemical substances produced by plants, but specific for some groups. Important physiological and ecological plant functions are attributed to flavonoids, such as photoprotection and pollinators attraction. For humans, this class of substances has medicinal properties, acting mainly as anti-inflammatory, antiviral, antitumor, antioxidant, hepatoprotective and antimicrobial. Croton L. is the second largest genus of Euphorbiaceae, with about 1300 species of trees, bushes or herbs. Croton is distributed in tropical and subtropical regions of the New and Old World, occurring in almost all Brazilian ecosystems. Croton cajucara is an example of species in clinical trials, having therapeutic indications for diabetes, hepatic and renal disorders, anti-inflammatory, antimicrobial, among others. Croton sphaerogynus Baill. emerges in the same clade of C. cajucara, and becomes a promising species for prospection of bioactive substances. This study aimed to investigate the phenolic composition and biological potential of phenolic substances present in leaf extracts of C. sphaerogynus and test their antioxidant, antimicrobial and anti - HIV potential. C. sphaerogynus was characterized as exclusive producer of flavonol, besides it is a major producer of diterpenes. Two flavonoids were isolated quercetin 3,7-dimethyl ether, and kaempferol 3-methyl ether. Regarding biological activities, the subfraction composed mainly of quercetin 3,7-dimethyl ether showed high antioxidant potential and the higher anti-HIV-1 activity when compared to extracts and other subfractions tested. The hexane fraction, rich in diterpenes, showed promising antimicrobial activity. Thus, C. spherogynus becomes a promising species for the uses of antioxidants substances

Thesis advisor: Kathleen McInnis-Dittrich / The purpose of this study was to investigate the dynamics of decision-making in formulating anti-poverty policies in Palestine. Particularly, this study was concerned with exploring the key decision makers, their roles, and how the power relationship among them influences the process of formulating anti-poverty policies. In addition, this study was intended to investigate the knowledge about the dynamics of decision making within the Palestinian National Authority (PNA): how actual decisions are made about anti-poverty policies and who is making decisions. Moreover, the scope of this study also included how the unique economic, social, and political contexts of Palestine influence the process of formulating anti-poverty policies. Further, this study explored how anti-poverty policies impact the lives of everyday poor Palestinians. This study utilized political theories, colonialism theory, and hegemony theory, to understand the external factors that affect the formulation of anti-poverty policies. Also, it used public policy theories, elitism, pluralism, and bounded rationality theory, to explore how anti-poverty policies are made and who made such policies in Palestine. This study employed a qualitative approach with a social constructivist paradigm of inquiry. This case study focused on two major sites that are responsible for formulating social policies in Palestine: the Ministry of Social Affairs (MOSA) and the Ministry of Planning and Administrative Development (MOPAD) in Ramallah. The findings of this study indicated that significant changes have occurred as regards who the key decision makers are and what roles they play in the formulation of anti-poverty policies. In analyzing the power relationship among the key decision makers, the findings showed that although the PNA has increased its control over the decision-making process, the international donor agencies continue to significantly influence this process. The data also revealed that unlike the models of policy making in democratic countries (such as elitism or pluralism), the approach to developing anti-poverty policies in Palestine reflects the participatory model. Consistent with the theory of bounded rationality, the findings revealed that anti-poverty policies have been made with financial, material, political, and other limitations. Implications for formulating anti-poverty policies and for future research are discussed. / Thesis (PhD) — Boston College, 2012. / Submitted to: Boston College. Graduate School of Social Work. / Discipline: Social Work.

Os parasitas nematóides afetam homens e animais, causando graves prejuízos à saúde pública e consideráveis perdas econômicas. A disponibilidade de anti-helmínticos de amplo espectro de ação tem auxiliado na redução de um grande número de perdas em decorrência das infestações parasitárias. Foram objetivos deste trabalho avaliar os efeitos do tratamento com moxidectina, na forma de longa-ação (LA), em sua dose terapêutica em touros, sobre a consistência e perímetro testicular, as características físicas (motilidade e vigor) e morfológicas do sêmen e o comportamento sexual. Foram utilizados 12 touros com idade de 48 ± 10 meses. Os animais foram alocados em 6 blocos com dois tratamentos. Os animais foram divididos nos grupos: controle (n=6), no qual cada animal recebeu 5 mL de solução fisiológica, via subcutânea, na orelha esquerda e grupo tratado (n=6), no qual cada animal recebeu 5 mL de moxidectina a 10% via subcutânea, na orelha esquerda. Os animais foram submetidos a exames andrológicos semanais, por um período de até 60 dias após o tratamento, sendo avaliadas as características testiculares (consistência e perímetro) e características seminais (motilidade, vigor e morfologia). Foram realizados testes de libido a cada 15 dias, num total de cinco testes. Os resultados obtidos indicam que não houve diferença significativa (P>0,05) para a consistência testicular, perímetro testicular, motilidade progressiva, vigor espermático, morfologia espermática e libido entre os touros dos grupos controle e tratado. Esses resultados indicam que o tratamento com moxidectina a 10% não influencia negativamente as características reprodutivas de touros. / The helminths affect human beings and animals, causing serious damage to public health and considerable economic losses. The availability of anthelminthic of a broad-spectrum has helped reducing a large number of losses due to parasitic infestation. The objectives of this work were to evaluate the effect of the treatment with moxidectin, in Long Action (LA) formulation, in therapeutic doses in bulls, on the testicular consistency and perimeter, physical characteristics (motility and strength) and morphological of the semen and sexual behaviour. Twelve bulls, age 48 ± 10 months used. These animals were put in six, 06 blocks with two treatments. The animals were divided in groups: Control group (n=6) each animal received 5 ml of physiological solution subcutaneously in the left ear, and Treated group (n=6) each animal received 5 ml of moxidectin at 10% subcutaneously in the right ear. The animals were submitted to andrological examinations weekly, within a period of 60 days after the treatment. The testicular characteristics (consistency and perimeter) and seminal characteristic (motility, strength and morphology) were evaluated. Libido tests were done every fortnight, in five tests all together. The results indicated that there were not significant differences. (P>0.05) for the testicular consistency, perimeter, progressive motility, spermatic strength, morphology and libido among bulls of the control and treated groups. These results indicated that the treatment with moxidectin at 10% does not influence negatively on the reproductive characteristics of the bulls.

GASPAROTTO, E.P.L. Expressão de genes e proteínas pró- e anti-apoptóticas em células precursoras da medula óssea e leucócitos do sangue periférico de pacientes portadores de policitemia vera. 2009. 185f. Tese de Doutorado. Faculdade de Ciências Farmacêuticas de Ribeirão Preto Universidade de São Paulo, Ribeirão Preto, 2009. A policitemia vera (PV) é uma doença mieloproliferativa clonal que afeta o progenitor hematopoético multipotente causando o acúmulo de eritrócitos, leucócitos e plaquetas morfologicamente normais e seus precursores, na ausência de um estímulo definido. Alterações na regulação da apoptose parece ser uma das causas do acúmulo de células eritróides sem a necessidade de eritropoetina. A mutação JAK2 V617F foi descrita como um dos eventos envolvidos na fisiopatologia das Doenças Mieloproliferativas Crônicas, sendo relacionada a um pior prognóstico. Esta mutação resulta na ativação constitutiva da enzima Janus Kinase 2 e conseqüentemente das vias de sinalização associadas ao estímulo celular mediado por citocinas e fatores de crescimento. O objetivo deste estudo foi investigar as alterações da expressão de proteínas e genes envolvidos na regulação da apoptose em pacientes com PV sem tratamento. Para tanto foi quantificada a expressão de genes e proteínas da família Bcl-2 e da via extrínseca da apoptose em células precursoras e leucócitos desses pacientes e de indivíduos saudáveis. Os leucócitos do sangue periférico de 12 pacientes e 14 controles foram obtidos pelo método de Haes-esteril. As células precursoras CD34+ destes pacientes e de 19 controles foram separadas em coluna imunomagnética. A detecção da expressão gênica e protéica das moléculas anti- e pró-apoptóticas foi realizada pelas técnicas de PCR em tempo real e Western-blot, respectivamente. Foi avaliada ainda a resistência à morte celular induzida por agentes apoptogênicos dos linfócitos dos pacientes. As células CD34+ e maduras mostraram expressão aumentada dos genes a1, mcl-1, noxa, c-flip e ciap-2. A expressão dos genes bid , bim-EL , fas e faim estava elevada e a dos genes bcl-2, bcl-xL e bax diminuída, somente nas células CD34+. Nos leucócitos, observou-se ainda maior nível de expressão do gene bok e expressão diminuída dos genes bcl-2, bcl-xL, bad, bax, faim, dr4, dr5 e trail. Os genes bcl-w, bak, bik, fasL e ciap-1 não mostraram diferença significativa na expressão em células CD34+ e leucócitos de pacientes e controles (p>0,05). Houve significância estatística nas correlações entre: expressão de dr5 e mutação JAK2 V617F; bim-EL com concentração de hemoglobina e com hematócrito; bcl-w e número de plaquetas; a1, bad, bax nas células CD34+ e bad em leucócitos com aumento do baço. Os linfócitos dos pacientes foram mais resistentes a apoptose mediada por ACT D 1uM e 5uM, ARA-C 25uM, CHX 25uM, VP-16 e VM-26 25uM. Houve correlação entre a percentagem de alelos mutados para JAK2 V617F e o perfil de resistência aos indutores de apoptose dos linfócitos dos pacientes a ACT D 1 uM e 5uM e STS 5uM. Observou-se ainda correlação estatística entre: expressão de a1 com ACT D 5 uM, CHX 50uM e ARA-C 25uM; bad com CHX 25uM e teniposídeo 25uM; bcl-2 com ACT D 5uM , CHX 50uM, STS 1uM e 5uM e VP-16 a25uM ; bid com ACT D 5uM e VM26 25uM; bik com ACT D 5uM , STS 1uM e 5uM e VM-26 25uM, ARAC 25uM e VP16 25Um; bim-EL com CHX 25uM e teniposídeo 25uM; bok com ACT D 5uM STS 1uM e 5uM, ARAC 25uM e etoposídeo 25uM; ciap-1 com ACT D 1uM e 5uM, STS 1uM e VP16 25uM; dr4 com ACT D 5uM, ARAC 25uM, VP16 a 25uM; dr5 com ACT D 5uM, STS 1uM, ARAC 25uM e VP16 25uM; faim com ACT D 5uM; mcl-1 com ARAC 25uM e STS 1uM; trail com ACT D 5uM. As proteínas A1, BAD e c-FLIP tiveram o mesmo perfil de expressão dos seus respectivos genes, contudo os níveis de expressão protéica para BCL-xL estavam aumentados e para BIM-EL diminuídos, resultados discordantes com os obtidos nas expressões gênicas. Em conjunto esses resultados sugerem que a maquinaria apoptótica está desregulada, o que poderia contribuir, pelo menos em parte, com a mieloacumulação observada no SP e MO dos pacientes com PV. / GASPAROTTO, E.P.L. Pro- and anti-apoptotic genes and proteins expression in bone marrow precursors and peripheral blood leukocytes in polycythemia vera patients. 185f. Thesis (Doctoral). Faculdade de Ciências Farmacêuticas de Ribeirão Preto Universidade de São Paulo, Ribeirão Preto, 2009. Polycythemia vera (PV) is a clonal myeloproliferative disease arising in a multipotent haematopoietic progenitor cell causing the accumulation of erythrocytes, leukocytes and platelets morphologically normal and their precursors in absence of a definable stimulus. Apoptosis deregulation seems to be one of the causes of erythroid cells accumulation without the need for erythropoietin. The JAK2 V617F mutation has been described as one of the events involved in pathophysiology of chronics myeloproliferative diseases, being related to worse prognosis. This mutation results in constitutive activation of the enzyme Janus kinase 2 and consequently of signaling pathways associated with cell stimulation mediated by cytokines and growth factors. This study aimed to investigate changes in expression of proteins and genes involved in regulation of apoptosis in PV patients without treatment. Was quantified for both gene and proteins expression of the Bcl-2 family and the extrinsic pathway of apoptosis members in precursor cells and leukocytes of patients and healthy subjects. Peripheral blood leukocytes of 12 patients and 14 controls were obtained by the Haes-sterile method. CD34+ precursor cells of these patients and 19 controls were separated by immunomagnetic column. Anti- and pro-apoptotic gene and protein expression detection was performed by real-time PCR and Western-blot techniques respectively. It also assessed the resistance to cellular death induced by apoptogenics agents in PV lymphocytes. CD34+ and mature cells showed increased expression of a1, mcl-1, noxa, c-flip and ciap-2. Gene expression of bid, bim-EL , fas e faim was high and to genes bcl-2, bcl-xL and bax diminished, only in CD34+ cells. In leukocytes, there was even greater level of expression of bok and reduced of bcl-2, bcl-xL, bad, bax, faim, dr4, dr5 and trail. Not significant difference was found to bcl-w, bak, bik, fasL and ciap-1 expression in CD34+ cells and leukocytes of patients and controls (p> 0.05). There was statistical correlation between: dr5 expression and JAK2 V617F mutation; bim-EL with hemoglobin concentration and hematócrito; bcl-w and platelets counts; a1, bad, bax in CD34+ cells and bad in leukocytes with enlarged spleen. PV lymphocytes were more resistant to apoptosis mediated by ACT D 1uM and 5uM, ARA-C 25uM, CHX 25uM, VP-16 25uM and VM-26 25uM. There was correlation between JAK2 V617F allele burden and resistance profile to apoptosis inducers in PV lymphocytes to ACT D 1uM and 5uM and STS 5uM. There was also correlation between: a1 expression with ACT D 5 uM, CHX 50uM and ARA-C 25uM; bad with CHX 25uM and teniposide 25uM; bcl-2 with ACT D 5uM, CHX 50uM, STS 1uM and 5uM and VP -16 25uM; bid with ACT D 5uM and VM-26 25uM; bik with ACT D 5uM, STS 1uM and 5uM, VM-26 25uM, ARA-C 25uM and VP-16 25uM; bim-EL with CHX 25uM and teniposide 25uM ; bok with ACT D 5uM, STS 1uM and 5uM, ARA-C 25uM and etoposide 25uM; ciap-1 with ACT D 1uM and 5uM, STS 1uM and VP-16 25uM; dr4 with ACT D 5uM, ARA-C 25uM, VP-16 25uM; dr5 with ACT D 5uM, STS 1uM, ARA-C 25uM and VP-16 25uM; faim with ACT D 5uM, mcl-1 with ARA-C 25uM and STS 1uM; trail with ACT D 5uM. The A1, BAD and c-FLIP proteins have the same profile of expression of their genes, however the levels of protein expression in BCL-xL were increased and decreased for BIM-EL, discordant results in related with the one obtained by gene expressions. Taken together these results suggest that apoptotic machinery is deregulated, which could contribute, at least in part, with myeloaccumulation observed in the SP and BM of PV patients.