EFAR - Escola de Farmácia

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O curso de Farmácia em Ouro Preto foi criado em 1839, sendo a mais antiga Escola de Farmácia da América Latina.

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Fase crônica cardíaca fibrosante da Tripanossomíase cruzi experimental no cão.
(1988) Lana, Marta de; Tafuri, Washington Luiz; Caliari, Marcelo Vidigal; Bambirra, Eduardo Alves; Chiari, Cléa de Andrade; Leite, Virginea Hora Rios; Barbosa, Alfredo José Afonso; Toledo, Max Jean de Ornelas; Chiari, Egler
De acordo com os trabalhos publicados ate o momento, o cão esta sendo considerado, com ressalvas, como modelo ideal para o estudo da fase aguda e crônica indeterminada da tripanossomiase cruz jl 2 3 4 5 6 7 14 15 18 19 20 21 24 Os requisitos para um modelo ideal, estabelecidos pelo Comite de Doenca de Chagas do Programa Especial de Treinamento e Pesquisa de Doenças Parasitarias da Organização Mundial de Saude25 podem ser assim discriminados: permitir o isolamento do parasito ao longo do curso da infecção; apresentar reações sorológicas positivas, indicativas da persistência da infecção; apresentar manifestações clinicas da doença de Chagas crônica; desenvolver miocardite, miosite e outras alterações patológicas que caracterizam a doença; induzir a resposta imune contra tecido do hospedeiro. Há mais de oito anos estamos a procura de um modelo que não somente preencha todos os requisitos acima citados mas, principalmente, que desenvolva a cardiopatia grave evolutiva fibrosante com todas alterações clinicas observadas na forma humana. Ate o momento, os resultados que encontramos parecem indicar que alcançamos tal objetivo no modelo cão. A partir destes resultados e dos de outros autores, tentaremos aplicar metodologia moderna no estudo dos vários fatores patogeneticos no pressuposto de que, assim, será possível chegar ao esclarecimento da patogenia e de fisiopatologia das diferentes formas anatomoclinicas da doença. Dentre os numerosos fatores patogeneticos ate agora aventados, a fibrose nos parece o mais importante na determinação da insuficiência cardíaca congestiva (ICC) e da aperistalse. Não existe qualquer outra cardiopatia e/ou mega com aspecto tão peculiar. No miocárdio bem como nos megas, a fibrose (fibrilopoese) e focal e difusa ao mesmo tempo23. O presente trabalho tem a finalidade de documentar a fase crônica da doença de Chagas em cães que recebem inóculos diversos das cepas Colombiana13 e Berenice-7817 de T. cruzi, destacando aqueles animais que desenvolveram a cardiopatia fibrosante, com sinais e sintomas clínicos de ICC.
Xanthones and Phenylcoumarins from Kielmeyera pumila.
(1988) Nagem, Tanus Jorge; Silva, Maurício de Abreu e
In continuation of our studies on extracts of the Guttiferae family we have examined the stems and fruit of Kielmeyera pumila Pohl, collected in the region of Ouro Preto, State of Minas Gerais, from a specimen identified by the botanist Jo& Badini (Herb. Prof. JosC Badini, Universidade Federal de Ouro Preto, Minas Gerais, Brazil). Several species of this genus has been shown to contain xanthones [l]. Investigation of the stems of the K. pumih has yielded, besides sitosterol, two previously known xanthones (osajaxanthone [Z] and 6-deoxyjacareubin [3]) and two known phenylcoumarins, the 1,2-dihydro-5- hydroxy-2-(l-hydroxy-l-methylethyl)-4-(3-methylbutyryl)- 6-phenyl-furo [2,3-h] [l] benzopyran-8-one [4] and mammeigin (1) [S]. The fruit of the same plant yielded friedelin, a-amyrin, shikimic acid, mammeigin and an isomer of mammeigin to which we have given the trivial name isomammeigin (2).
Hemocultures for the parasitological diagnosis of human chronic Chagas' disease.
(1989) Chiari, Egler; Dias, João Carlos Pinto; Lana, Marta de; Chiari, Cléa de Andrade
With the purpose of standardization of an hemoculture technique presenting a higher positive rate in the parasitological diagnosis of chronic Chagas’ disease in patients with reactive serology (IFT, HA, CFT) the following schedule was used. Thirty ml of venous blood was collected with heparin and the plasma was separated by centrifugation (2.000 rpm/30'). The packed cells were washed with LIT medium or PBS which was then removed by centrifugation (2.000 rpm/15’). This material was sampled in 6 screw-tubes 18x200 with 6 ml of LIT medium and incubated at 28°C. These incubated cultures at 28°C were examined after 15, 30, 45 and 60 days. When the hemoculture was not immediately processed after blood collection, the plasma was removed and the sediment enriched with LIT medium and preserved at 4°C. The Xenodiagnosis was performed according to Schenones method used here as a reference technique. Among the various groups of patients examined by both techniques the best results obtained were: 55.08% ofpositivity for hemocultures against 27.5% forxenodiagnosis (X^ = 4.54, p = 0.05), with a tubepositivity of 26.6%. Recommendation for screening trials of drug assays is the repetition of method on a same patient 2 or more times in different occasions, as used in xenodiagnosis.
Regulation of sugar transport systems in Fusarium oxysporum var. lini.
(1990) Brandão, Rogélio Lopes; Dias, Maria C. Loureiro
Fusarium oxysporum var. Iini (ATCC 10960) formed a facilitated diffusion system for glucose (Ks, about 10 mM) when grown under repressed conditions. Under conditions of derepression, the same system was present together with a high-affinity (K5, about 40 ,iM) active system. The maximum velocity of the latter was about 5% of that of the facilitated diffusion system. The high-affinity system was under the control of glucose repression and glucose inactivation. When lactose was the only carbon source in the medium, a facilitated diffusion system for lactose was found (Ks, about 30 mM).
Humoral immune response in dogs experimentally infected with Trypanosoma cruzi.
(1991) Lana, Marta de; Vieira, Lauro Mello; Coelho, George Luiz Lins Machado; Chiari, Egler; Veloso, Vanja Maria; Tafuri, Washington Luiz
Leishmaniose visceral canina : avaliação da metodologia sorológica utilizada em inquéritos epidemiológicos.
(1991) Costa, Carlos Alberto da; Genaro, Odair; Lana, Marta de; Magalhães, Paulo Araújo; Dias, Magno; Michalick, Marilene Susan Marques; Melo, Maria Norma; Costa, Roberto Teodoro da; Rocha, Neuza Maria de Magalhaes; Mayrink, Wilson
Foi realizado um estudo comparativo da reação de imunofluorescencia indireta em eluatos de sangue de cães infectados experimentalmente com diferentes tripanosomatideos. Utilizaram-se como antigenopromastigotas de L. mexicana, L. braziliensis e L. chagasi. Os resultados mostraram que a sensibilidade do método foi de 87,5% para o diagnostico do calazar canino, independentemente do antígeno empregado; e que ocorre reação cruzada com Leishmaniose tegumentar em 75% dos casos e com doença de Chagas em 83,3%. Levantamento epidemiológico em área de leishmaniose confirma que a reação de imunofluorescencia em eluatos de sangue canino fornece reações cruzadas em cães infectados com Leishmania braziliensis e L. chagasi. Não se verificou reação cruzada pela RFC. Sugere-se a utilização da reação de imunofluorescencia nas campanhas de saúde publica, mas e de se chamar a atenção para o fato de que as taxas de positividade não devem ser utilizadas como indicadores da prevalência do calazar canino.
Effect of substrate and pH on the activity of proteases from Fusarium oxysporum var. lini.
(1991) Castro, Ieso de Miranda; Lima, Angélica Alves; Paula, Carmem Aparecida de; Nicoli, Jacques Robert; Brandão, Rogélio Lopes
The results obtained in this work suggest that both the pH (through selective inhibition) and the carbon source (through repression and acidification or alkalinization of the medium) may play an important role in the distribution of extracellular proteases in Fusarium oxysporum var. lini.
Glycerol utilization in Fusarium oxysporum var lini : regulation of transport and metabolism.
(1991) Castro, Ieso de Miranda; Dias, Maria C. Loureiro
Glycerol was transported in the fungus Fusarium oxysporum var. lini by a facilitated diffusion transport system with a half-saturation constant, Ks, of 0.5 mM and a maximum velocity, Vmax, of 0.9 mmol (g dry wt)-l h-l at pH 5 and 25OC. 1,2-Propanediol was a competitive inhibitor of glycerol transport, but the cells did not actively accumulate 1,2-propanediol. The transport system was partially constitutive. In cells grown in the presence of glucose, glycerol was not transported, indicating that the synthesis of the system was under glucose repression. Glycerol base and NADP+-dependent glycerol dehydrogenase activities were present under all physiological conditions tested. A flavin-dependent glycerol phosphate dehydrogenase was induced only when glycerol was the sole energy source in the medium. This enzyme, together with the transport system, constitute the regulated steps in the glycerol metabolic pathway.
Experimental Chagas' disease in dogs.
(1992) Lana, Marta de; Chiari, Egler; Tafuri, Washington Luiz
Glucose-induced activation of plasma membrane H+-ATPase in mutants of the yeast Saccharomyces cerevisiae affected in cAMP metabolism, cAMP-dependent protein phosphorylation and the initiation of glycolysis.
(1992) Passos, Jomar Becher dos; Vanhalewyn, Mieke; Brandão, Rogélio Lopes; Castro, Ieso de Miranda; Nicoli, Jacques Robert; Thevelein, Johan Maria
Addition of glucose-related fermentable sugars or pro,tonophores to derepressed cells of the yeast Saccharomyces ceret'isiae causes a 3- to 4-fold activation of the plasma membrane H +-A'fPase within a few minutes. These conditions are known to cause rapid increases in the cAMP level. In yeast strains carrying temperature-sensitive mutations in genes required for cAMP .~jnthesis, incohati~a at the restrictive temperature reduced the extent of H+-ATPase activation, Incubation of nontemperature- sensitive strains, however, at such temperatures also caused reduction of H +-ATPase activation. Yeast strains which are specifically deficient in the glucose-induced cAMP increase (and not in basal cAMP synthesis) still showed plasma membrane H+-ATPase aCtivation. Yeast mutants with widely divergent activity levels of cAMP-dependent protein kinase displayed very similar levels of activation of the plasma membrane H +-A'l'Pase. This was also true for a yeast mutant carrying a deletion in the CDC25 gene. These results show that the cAlVlP-protein kinase A signaling pathway is not required for glucose activation of the H*-ATPase. They also contradict the specific requirement of the CDC25 gene product. Experiments with yeast strains carrying point or deletion mutations in the genes coding for the sugar phnsphorylating enzymes hexokinase Pl and Pll and glucokinase showed that activation of the H+-ATPase with glucose or fructose was completely dependent on the presence cf a kinase able m phnsphorylate the sugar. These and other data concerning the role of init,:al sugar metabolism in triggering activation are consistent with the idea that the glucose-induced activation pathways of cAMP-synthesis and H+-ATPase have a common initiation point.
Glucose induced activation of the plasma membrane ATPase in Fusarium oxysporum.
(1992) Brandão, Rogélio Lopes; Castro, Ieso de Miranda; Passos, Jomar Becher dos; Nicoli, Jacques Robert; Thevelein, Johan Maria
Addition of glucose and other sugars to derepressed cells of the fungus Fusarium oxysporum var. Zini triggered activation of the plasma membrane H+-ATPase within 5 min. Glucose was the best activator while galactose and lactose had a lesser effect. The activation was not prevented by previous addition of cycloheximide and it was fully reversible when the glucose was removed. The activation process in uiuo also caused changes in the kinetic properties of the enzyme. The non-activated enzyme had an apparent K, of about 3.2 mM for ATP whereas the activated enzyme showed an apparent K,,, of 0.26 mM. In addition, the pH optimum of the H+-ATPase changed from 6.0 to 7.5 upon activation. The activated enzyme was more sensitive to inhibition by vanadate. When F. oxysporum was cultivated in media containing glucose as the major carbon source, enhanced M+-ATPase activity was largely confined to the period corresponding to the lag phase, i.e. just before the start of acidification of the medium. This suggests that the activation process might play a role in the onset of extracellular acidification. Addition of glucose to F. oxysporum var. Zini cells also caused an increase in the cAMP level. No reliable increase could be demonstrated for the other sugars. Addition of proton ionophores such as DNP and CCCP at pH 5-0 caused both a large increase in the intracellular level of cAMP and in the activity of the plasma membrane H+- ATPase. Inhibition of the DNP-induced increase in the cAMP level by acridine orange also resulted in inhibition of the activation of plasma membrane H+-ATPase. These results suggest a possible causal relationship between the activity of F. oxysporum var. Zini plasma membrane H+-ATPase and the intracellular level of CAMP.
Molecular cloning of a gene involved in glucose sensing in the yeast Saccharomyces cerevisiae.
(1993) Aelst, Linda Van; Hohmann, Stefan; Bulaya, Botchaka; Koning, Wim de; Sierkstra, Laurens; Neves, Maria José; Luyten, Kattie; Alijo, Rafael; Ramos, José; Coccetti, Paola; Martegani, Enzo; Rocha, Neuza Maria de Magalhaes; Brandão, Rogélio Lopes; Dijck, Patrick Van; Vanhalewyn, Mieke; Durnez, Peter; Jans, Arnold W. H; Thevelein, Johan Maria
Cells of the yeast Saccharomyces cerevisiae display a wide range of glucose-induced regulatory phenomena, including glucose-induced activation of the RAS-adenylate cyclase pathway and phosphatidylinosrtot turnover, rapid post-translatronal effects on the activity of different enzymes as well as long-term effects at the transcriptional level. A gene called GGS1 (for general Glucose Sensor) that is apparently required for the glucose-induced regulatory effects and several ggsi aHeles (fic/pf, bypi and cifi) has been cloned and characterized. A GGS1 homologue is present in Methanobacterium thermoautotrophicum. Yeast ggsi mutants are unable to grow on glucose or Received 25 November, 1992; revised and accepted 15 February, 1993. •For correspondence. Tel. (16) 220931; Fax (16) 204415. tThese two authors contributed equally to this paper. related readily fermentable sugars, apparently owing to unrestricted influx of sugar Into glycolysis, resulting in its rapid deregulation. Levels of intracellular free glucose and metabolites measured over a period of a few minutes after addition of glucose to cells of a ggsi^ strain are consistent with our previous suggestion of a functional interaction between a sugar transporter, a sugar kinase and the GGS1 gene product. Such a glucose-sensing system might both restrict the influx of glucose and activate several signal transduction pathways, leading to the wide range of glucose-induced regulatory phenomena. Deregulation of these pathways in ggsi mutants might explain phenotypic defects observed in the absence of glucose, e.g. the inability of ggsi diploids to sporulate.
Possible involvement of a phosphatidylinositol-type signaling pathway in glucose-induced activation of plasma membrane ATPase and cellular proton in the yeast Sacchamyces cerevisiae.
(1994) Brandão, Rogélio Lopes; Rocha, Neuza Maria de Magalhaes; Alijo, Rafael; Ramos, José; Thevelein, Johan Maria
Addition of glucose to cells of the yeast Saccharomyces cerevisiae causes rapid activation of plasma membrane H+-ATPase and a stimulation of cellular H ÷ extrusion. We show that addition of diacylglycerol and other activators of protein kinase C to intact cells also activates the H+-ATPase and causes at the same time a stimulation of H ÷ extrusion from the cells. Both effects are reversed by addition of staurosporine, a protein kinase C inhibitor. Addition of staurosporine or calmidazolium, an inhibitor of Ca2+/calmodulin-dependent protein kinases, separately, causes a partial inhibition of glucose-induced H+-ATPase activation and stimulation of cellular H + extrusion; together they cause a more potent inhibition. Addition of neomycin, which complexes with phosphatidylinositol 4,5-bisphosphate, or addition of compound 48/80, a phospholipase C inhibitor, also causes near complete inhibition. Diacylglycerol and other protein kinase C activators had no effect on the activity of the K+-uptake system and the activity of trehalase and glucose-induced activation of the K+-uptake system and trehalase was not inhibited by neomycin, supporting the specificity of the effects observed on the H+-ATPase. The results support a model in which glucose-induced activation of H+-ATPase is mediated by a phosphatidylinositol-type signaling pathway triggering phosphorylation of the enzyme both by protein kinase C and one or more Ca2+/calmodulin-dependent protein kinases.
Utilization of lactic acid by Fusarium oxysporum var. lini : regulation of transport and metabolism.
(1994) Castro, Ieso de Miranda; Dias, Maria C. Loureiro
Lactic acid was transported in Fusarium oxysporum var. lini ATCC 10960 by a saturable transport system that had a half-saturation constant of 56.6 7.5 FLM and a maximum velocity of 0.61 0.10 mmol h-1 g-1 (dry weight) at 26°C and pH 5.0. This transport system was inducible and was not expressed in the presence of a repressing substrate. Evidence is presented that the anionic form lactate- was taken up by the cells. Propionic, acetic, pyruvic, and bromoacetic acids but not succinic acid competitively inhibited the transport of lactic acid. Bromoacetic acid, which was not metabolized, was taken up to a steady-state level when intracellular and extracellular concentrations were identical, indicating that the transport system was not accumulative. The enzymatic activity that was physiologically more relevant in the metabolism of lactic acid was lactate: ferricytochrome c oxidase. This enzyme did not exhibit stereospecifity and was induced by lactic acid.
Granulomatous hypersensitivity to Schistosoma mansoni egg antigens in human schistosomiasis. IV. A role for prostaglandin-induced inhibition of in vitro granuloma formation.
(1994) Goes, Alfredo Miranda de; Rezende, Simone Aparecida; Gazzinelli, Giovanni; Doughty, Barbara L.
The prostaglandins (PG) are known to regulate immune cell function (s) and participate in the progression of both acute and chronic inflammatory reactions. Using an in vitro model of Schistosoma mansoni egg-induced hypersensitivity granulomas, we have delineated the role of immune complexes (IC) in the induction andrelease of PG and their inhibitory effects on granuloma development. The hypersensitivity- type granuloma reaction to soluble egg antigen (SEA) was examined using a model of in vitro granuloma ,formation. Our results show that granuloma formation was dramatically suppressed by the addition to the granuloma cultures of IC, PGE,, PGE2, while PGF, alpha had no significant effect. The inhibition of the PG function was achieved by the introduction of anti-PG antibodies that blocked suppression of granuloma,formation. It appears in this model system that IC may inhibit the activity of granuloma formation by stimulating the monocyte-macrophage lineage to release inhibitory mediators. Our results suggest that the prostaglandins E series may be important in the generation and maintenance of suppression of the granulomatous inflammatory response to S. mansoni egg antigens .
Maillard reaction during the processing of "Doce de Leite".
(1994) Pavlovic, Suzana; Santos, Rinaldo Cardoso dos; Glória, Maria Beatriz Abreu
‘Doce de leite’, a dairy product widely consumed in Brazil as a dessert or cake filling, is obtained from the heat treatment of milk and sucrose. On heating, the Maillard reaction occurs with the formation of desirable browncoloured products with a characteristic and pleasant flavour. However, the reaction can also lead to changes in nutritive value. In order to follow chemically the extension of the Maillard reaction and its effect on the nutritive value of ‘doce de leite’ its commercial processing was followed. Increases in the level of free 5-hydroxymethylfurfural and in absorption at 420 nm by pigments attached to the protein were observed. The amino acid analysis showed a significant decrease in lysine levels (33%) as well as in arginine (11%) and histidine (10%). There was also a reduction in available lysine levels, as measured by the fluorodinitrobenzene (50.6%) and by the 2,4,6-trinitrobenzenesulphonic acid (23.5%) methods. These results suggest a deleterious effect of the processing of ‘doce de leite’ on the nutritive value of the milk.
Cardiac plexus of dogs experimentally infected with Trypanosoma cruzi: inflammatory lesions and quantitative studies.
(1995) Caliari, Marcelo Vidigal; Lana, Marta de; Caliari, Estela Regina de Oliveira; Tafuri, Washington Luiz
Qualitative and quantitative aspects of the superficial and profound cardiac plexus of dogs experimentally infected with Be-62 and Be-78 strains of Trypanosoma cruzi were studied. Animals were autopsied in the acute phase o f infection. The inflammatory process, lesions and number of parasites were more intense and frequent in animals infected with the Be-78 strain than in those infected with Be-62. Despite this, no statistically significant differences could be found between the number of neuron bodies in the ganglia of infected and control dogs.
Cell surface analysis of trypanosomes of the subgenus Schizotrypanum isolate from bats.
(1996) Pinto, Artur da Silveira; Teixeira, Luiz Fernando de Medeiros; Padron, Thais Cristina Baeta Soares Souto; Andrade, Arnaldo Feitosa Braga de
Two stocks (M5, M29) of trypanosomes of the subgenus Schizotrypanum were isolated from the bat Phylloslomus hastatus and analyzed for cell electrophoretic mobility (EPM) and lectin binding surface sites. Epimastigotes from the M5 and M29 stocks presented a mean EPM of around - 0.57 and -0.56 μm, s-1.V-1.cm, respectively. Differences in the agglutination profiles were detected between epimastigotes or trypomastigotes from the two parasite populations using lectins with specificity for D-GlcNAc, D-GalNAc, D-Gal and D-Man as probe. Major variation was observed between epimastigote forms. Additionally, the D-GlcNAc binding lectins WGA and BS II strongly interacted with the trypomastigote from both M5 and M29 stocks; this fact is evidence that these trypanosomes are distinct from Trypanosoma (Schizotrypanum) cruzi.
Estudo quantitativo e qualitativo dos plexos de Auerbach e Neissner do esôfago de cães inoculados com o Trypanosoma cruzi.
(1996) Caliari, Estela Regina de Oliveira; Caliari, Marcelo Vidigal; Lana, Marta de; Tafuri, Washington Luiz
Andrade caracterizou bem a fase aguda e a forma indeterminada da cardiopatia chagásica no cão1. Lana inoculando a cepa Be78 em cães caracterizou muito bem a cardiopatia chagásica crônica fibrosame, com quadro clínico e eletrocardiográfico muito semelhante a forma cardíaca humana". Pelo que sabemos, até o momento, não se documentou claramente a forma digestiva (megaesôfago e megacólon) experimental. Também não se tem notícias de estudos sistematizados dos neurônios dos plexos de Auerbach e Meissner do esôfago na tripanosomía s e cruzi experimental. Por estas razões realizamos o presente trabalho , no pressuposto de encontrar prováveis lesões neuronais do esôfago na infecção chagásica experimental canina.
Characterization of two isolates of Trypanosoma cruzi obtained from the patient Berenice, the first human case of Chagas disease.
(1996) Lana, Marta de; Chiari, Cléa de Andrade; Chiari, Egler; Morel, Carlos Medicis; Gonçalves, Antônio M.; Romanha, Alvaro José
Two isolates of Trypanosoma cruzi were obtained from the patient Berenice, the first human case of Chagas’ disease (Chagas 1909), when she was 55 and 71 years old, respectively. The isolates were characterized on the basis of their epimastigote-trypomastigote differentiation in liquid media and of the electrophoretic pattern of EcoR1 digestion products of kinetoplast DNA (k- DNA) minicircles (schizodeme) and isoenzyme patterns (zymodeme). Clear differences were found between the isolates, suggesting the occurrence of a heterogeneous population of T. cruzi in the infection of this patient.

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