RESUMO
Canine cutaneous leishmaniasis (CCL) is an emerging zoonotic infection endemic in several countries of the world. Due to variable response to therapy and frequency of relapses, a more effective, safer, and inexpensive treatment is needed. Recently, it was reported that the hederagenin glucoside saponins (SS) and chromane-derived hydrazone (TC2) combined in a 1:1 ratio has high potential in antileishmanial therapy since both compounds alter the survival of Leishmania and the ability to infect adjacent macrophage. Not only the skin permeation and the absorption of an ointment containing 2% TC2 and 2% SS (w/w) was determined in this work, but also the acute dermal toxicity in both in vitro and in vivo assays. Last, the effectiveness and safety of the topical therapy with 2% TC2-2% SS ointment was evaluated in an observational study in dogs with diagnosis of cutaneous leishmaniasis (CL). Both TC2 and SS diffused through pig ear skin and traces of TC2 (but not SS) were detected in the stratum corneum of mice at 6-24 h. Neither TC2 nor SS was detected in plasma. The acute dermal toxicity was negative. Treatment with 2% TC2-2% SS ointment produced a complete long-term clinical cure in 56 dogs (24 females and 32 males) from the Orinoco and Amazonas regions in southeastern Colombia without adverse effects. All dogs have remained disease-free for the last 24 months. In conclusion, these results support the use of this topical therapy as a safer and new first-line local treatment of CCL that could help limit the spread of CL from dogs to humans.
Assuntos
Antiprotozoários , Leishmaniose Cutânea , Saponinas , Animais , Antiprotozoários/uso terapêutico , Cães , Feminino , Glucosídeos/uso terapêutico , Hidrazonas/uso terapêutico , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/veterinária , Masculino , Camundongos , Pomadas/uso terapêutico , Ácido Oleanólico/análogos & derivados , Saponinas/uso terapêutico , SuínosRESUMO
T. rangeli epimastigotes contain only a single detectable phosphoglycerate kinase (PGK) enzyme in their cytosol. Analysis of this parasite's recently sequenced genome showed a gene predicted to code for a PGK with the same molecular mass as the natural enzyme, and with a cytosolic localization as well. In this work, we have partially purified the natural PGK from T. rangeli epimastigotes. Furthermore, we cloned the predicted PGK gene and expressed it as a recombinant active enzyme. Both purified enzymes were kinetically characterized and displayed similar substrate affinities, with KmATP values of 0.13â¯mM and 0.5â¯mM, and Km3PGA values of 0.28â¯mM and 0.71â¯mM, for the natural and recombinant enzyme, respectively. The optimal pH for activity of both enzymes was in the range of 8-10. Like other PGKs, TrPGK is monomeric with a molecular mass of approximately 44â¯kDa. The enzyme's kinetic characteristics are comparable with those of cytosolic PGK isoforms from related trypanosomatid species, indicating that, most likely, this enzyme is equivalent with the PGKB that is responsible for generating ATP in the cytosol of other trypanosomatids. This is the first report of a glycolytic enzyme characterization from T. rangeli.
Assuntos
Fosfoglicerato Quinase/genética , Trypanosoma rangeli/enzimologia , Sequência de Aminoácidos , Sequência de Bases , Cromatografia em Gel , Cromatografia por Troca Iônica , Clonagem Molecular , Sequência Consenso , Citosol/enzimologia , DNA Intergênico/química , Concentração de Íons de Hidrogênio , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/isolamento & purificação , Isoenzimas/metabolismo , Cinética , Fosfoglicerato Quinase/química , Fosfoglicerato Quinase/isolamento & purificação , Fosfoglicerato Quinase/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Trypanosoma rangeli/genéticaRESUMO
In this paper, we report the subcellular distribution of phosphoglycerate kinase (PGK) in epimastigotes of Trypanosoma cruzi. Approximately 80% of the PGK activity was found in the cytosol, 20% in the glycosomes. Western blot analysis suggested that two isoenzymes of 56 and 48 kDa, respectively, are responsible for the glycosomal PGK activity, whereas the cytosolic activity should be attributed to a single PGK of 48 kDa. In analogy to the situation previously reported for PGK in Trypanosoma brucei, these isoenzymes were called PGKA, C and B, respectively. However, in T. cruzi, PGKA seems not to be a minor enzyme like its counterpart in T. brucei. Whereas PGKC behaved as a soluble glycosomal matrix protein, PGKA appeared to be present at the inner surface of the organelle's membrane. After alkaline carbonate treatment, the enzyme remained associated with the particulate fraction of the organelles. Upon solubilization of glycosomes with Triton X-114, PGKA was recovered from the detergent phase, indicating its (partial) hydrophobic character and therefore, a possible hydrophobic interaction with the membrane. The PGKA gene was cloned and sequenced, but the predicted amino-acid sequence did not reveal an obvious clue as to the mechanism by which the enzyme is attached to the glycosomal membrane.
Assuntos
Fosfoglicerato Quinase/metabolismo , Frações Subcelulares/enzimologia , Trypanosoma cruzi/enzimologia , Sequência de Aminoácidos , Animais , Citosol/enzimologia , Isoenzimas/metabolismo , Microcorpos/enzimologia , Dados de Sequência Molecular , Fosfoglicerato Quinase/genética , Análise de Sequência de DNA , Trypanosoma cruzi/genética , Trypanosoma cruzi/crescimento & desenvolvimentoRESUMO
alpha-glycerophosphate dehydrogenase (alpha-GPDH-EC.1.1.1.8) has been considered absent in Trypanosoma cruzi in contradiction with all other studied trypanosomatids. After observing that the sole malate dehydrogenase can not maintain the intraglycosomal redox balance, GPDH activity was looked for and found, although in very variable levels, in epimastigotes extracts. GPDH was shown to be exclusively located in the glycosome of T. cruzi by digitonin treatment and isopycnic centrifugation. Antibody against T. brucei GPDH showed that this enzyme seemed to be present in an essentially inactive form at the beginning of the epimastigotes growth. GPDH is apparently linked to a salicylhydroxmic-sensitive glycerophosphate reoxidizing system and plays an essential role in the glycosome redox balance.
Assuntos
Glicerolfosfato Desidrogenase/análise , Microcorpos/química , Proteínas de Protozoários/análise , Trypanosoma cruzi/química , Animais , Glicerolfosfato Desidrogenase/metabolismo , Microcorpos/enzimologia , Consumo de Oxigênio , Proteínas de Protozoários/metabolismo , Trypanosoma cruzi/enzimologiaRESUMO
alpha-glycerophosphate dehydrogenase (alpha-GPDH-EC.1.1.1.8) has been considered absent in Trypanosoma cruzi in contradiction with all other studied trypanosomatids. After observing that the sole malate dehydrogenase can not maintain the intraglycosomal redox balance, GPDH activity was looked for and found, although in very variable levels, in epimastigotes extracts. GPDH was shown to be exclusively located in the glycosome of T. cruzi by digitonin treatment and isopycnic centrifugation. Antibody against T. brucei GPDH showed that this enzyme seemed to be present in an essentially inactive form at the beginning of the epimastigotes growth. GPDH is apparently linked to a salicylhydroxmic-sensitive glycerophosphate reoxidizing system and plays an essential role in the glycosome redox balance
Assuntos
Animais , Glicerolfosfato Desidrogenase/análise , Microcorpos/química , Trypanosoma cruzi/química , Glicerolfosfato Desidrogenase/metabolismo , Microcorpos/enzimologia , Consumo de Oxigênio , Trypanosoma cruzi/enzimologia , Trypanosoma cruzi/metabolismoRESUMO
Three polyketides alpha-pyrones, named passifloricins, were isolated from Passiflora foetida resin; their structures and relative configurations were assigned through 2D NMR spectroscopic analyses. These types of compounds were not detected in other passion flowers.
Assuntos
Pironas/isolamento & purificação , Rosales/química , Espectroscopia de Ressonância Magnética , Folhas de Planta/química , Pironas/químicaRESUMO
Fascioliasis is a cosmopolitan parasitic disease. The development of some diagnostic imaging methods such as ultrasonography (US) and endoscopic retrograde cholangio-pancreatography (ERCP) have provided the opportunity of detecting the presence of the parasite in the gall-bladder and in the extra-hepatic biliary tract. Four cases of fascioliasis in the biliary tract are shown, in which the ultrasonographic findings suggested a diagnosis of fascioliasis which was later confirmed by biliary drainage and/or stool culture. When there is no clinical evidence of parasitism, ultrasonography may help to assume the presence of the parasite. The ERCP is useful to confirm the location of the trematode in the extra-hepatic biliary tract. Biliary tract fascioliasis may present as a cholestatic syndrome.
Assuntos
Doenças dos Ductos Biliares/diagnóstico por imagem , Fasciolíase/diagnóstico por imagem , Adulto , Doenças dos Ductos Biliares/parasitologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Radiografia , UltrassonografiaRESUMO
A new phytoalexin was induced and isolated from papaya fruit slices treated with copper salts; its structure was established as 3',5'-dimethoxy-4'-hydroxy-(2-hydroxy)acetophenone. This compound showed high antifungal activity against Colletotrichum gloesporioides, a pathogenic fungus of papaya.