Tarsila Ferraz Frezza

Hypoxia, hypoxia-inducible factor-1α and vascular endothelial growth factor in a murine model of Schistosoma mansoni infection

Schistosomiasis mansoni is a chronic parasitic disease where much of the symptomatology is attributed to granuloma formation, an immunopathological reaction against Schistosoma eggs. To more clearly understand the immunopathology of schistosomiasis, the tissue microenvironment generated by S. mansoni infected mice was investigated. Using the hypoxia marker pimonidazole, we provide immunohistochemical evidence that hypoxia occurred in inflammatory cells infiltrated around the eggs and cells surrounding granulomas in the liver, intestine, spleen and lungs of infected mice. Hypoxia-inducible factor-1α (HIF-1α) was mainly expressed in inflammatory cells surrounding the eggs and in hepatocytes surrounding cellular and fibrocellular granulomas in infected mouse liver. HIF-1α expression was also verified in granulomas in the other tissues tested (intestine, spleen and lungs). Vascular endothelial growth factor (VEGF) expression was observed in the extracellular space surrounding inflammatory cells in liver granuloma. The VEGF expression pattern verified in infected mouse liver was very similar to that observed in the other tissues tested. A strong positive correlation occurred between pimonidazole binding and HIF-1α and VEGF expression in the tissues tested, except for lung. This work is the first evidence that infection by a helminth parasite, S. mansoni, produces a hypoxic tissue microenvironment and induces HIF-1α and VEGF expression.

Liposomal-praziquantel: efficacy against Schistosoma mansoni in a preclinical assay.

Currently, schistosomiasis mansoni is treated clinically with praziquantel (PZQ). Nevertheless, cases of tolerance and resistance to this drug have been reported, creating the need to develop new drugs or to improve existing drugs. Considering the small number of new drugs against Schistosoma mansoni, the design of nanotechnology-based drug delivery systems is an important strategy in combating this disease. The aim of this study was to evaluate the activity of PZQ containing liposome (lip.PZQ) on S. mansoni, BH strain. Mice were treated orally with different concentrations of PZQ and lip.PZQ 30 and 45 days following infection. The number of worms, recovered by perfusion of the hepatic portal system, and the number of eggs found in the intestine and liver were analysed. Parasite egg counts were also performed. The most active formulation for all parameters was 300mg/kg of lip.PZQ, since as it decreased the total number of worms by 68.8%, the number of eggs in the intestine by 79%, and the number of hepatic granulomas by 98.4% compared to untreated controls. In addition, this concentration decreased egg counts by 55.5%. The improved efficacy of the treatment with lip.PZQ, especially when administered 45 days following infection, compared with the positive-control group (untreated) and the groups that received free PZQ, can be explained by greater bioavailability in the host organism; the preferred target of lip.PZQ is the liver, and lip.PZQ is better absorbed by the tegument of S. mansoni, which has an affinity for phospholipids.

The Use of Brazilian Medicinal Plants to Combat Schistosoma mansoni

Human schistosomiasis, which is caused by trematodes of the genus Schistosoma, is considered one of the most prevalent and debilitating neglected diseases in tropical and subtropical areas, being endemic to approximately 76 countries and territories and affecting more than 207 million people worldwide. About 800 million people live in infection risk areas and 280,000 die every year due to this disease (Engels et al., 2002, Steinmann et al., 2006). There are six main species of Schistosoma that can infect humans: S. mansoni, S. hematobium, S. japonicum, S. intercalatum, S. mekongi and S. malayensis. Among these, only S. mansoni is found in Brazil. S. mansoni schistosomiasis was probably brought to Brazil by the Atlantic slave trade. Afterwards, migration flows spread the disease from the shore to the interior of the country, including areas containing its intermediate hosts, planorbides of the genus Biomphalaria (Paraense, 1986, Coura & Amaral, 2004). According to Paraense (1966, 1981), ten species and one subspecies of Biomphalaria can be found in the country, three of them being natural intermediate hosts (B. glabrata, B. tenagophila, B. straminea) and two having already been experimentally infected (B. amazonica and B. peregrina). The life cycle of S. mansoni consists of an asexual reproduction stage in the intermediate hosts and a sexual reproduction stage in the definite ones. Under favourable conditions, it lasts 80 days (Katz & Almeida, 2003, Ministerio da Saude, 2005). Nowadays, there are 25 million people living in endemic areas in Brazil, and 4 to 6 million are infected. It is the most affected country in the Americas (Lambertucci, 2010, World Health Organization [WHO], 2010). According to data provided by Brazil’s Ministry of Health, there are cases of the disease in 19 states, from Rio Grande do Norte to Bahia and in the interior of Espirito Santo and Minas Gerais (Ministerio da Saude, 2005). Figure 1 shows the distribution of S. mansoni schistosomiasis in Brazil. A major problem in countries where schistosomiasis is endemic is the control of the disease. In this regard, isolated or combined measures could be taken, such as the control of the intermediate hosts using molluscicides, improvement of basic sanitation and water supply conditions, sanitary education of exposed populations, individual or mass treatment in

Effectiveness of hyperbaric oxygen for experimental treatment of schistosomiasis mansoni using praziquantel-free and encapsulated into liposomes: assay in adult worms and oviposition.

The treatment of schistosomiasis depends on a single drug: praziquantel (PZQ). However, this treatment presents limitations such as low and/or erratic bioavailability that can contribute to cases of tolerance. Improvements to the available drug are urgently needed and studies with a controlled system of drug release, like liposomes, have been gaining prominence. The present study evaluated the activity and synergy between liposomal-praziquantel (lip.PZQ) and hyperbaric oxygen therapy (HBO). Mice received doses of 60 or 100mg/kg PZQ or lip.PZQ, 50 days post-infection, and after the treatment, were exposed to HBO (3 atmosphere absolute - ATA) for 1h. The viability of adult worms and oviposition were analyzed, by necropsy and Kato-Katz examination performed after 15 days of treatment. A concentration of 100mg/kg of lip.PZQ+HBO was more effective (48.0% reduction of worms, 83.3% reduction of eggs/gram of feces) and 100% of the mice had altered of oograms (indicating interruption of oviposition) compared to other treatments and to the Control group (infected and untreated). It is known that PZQ requires participation of the host immune system to complete its antischistosomal activity and that HBO is able to stimulate the immune system. The drug became more available in the body when incorporated into liposomes and, used with HBO, the HBO worked as an adjuvant. This explains the decreases of oviposition and worms recovered form hepatic portal system.

Copépodos Notodiaptomus sp. Kiefer (Crustacea, Calanoida) naturalmente infectados com metacestódeos no reservatório do Juqueri, São Paulo, Brasil

The aim of this work was to identify the components of zooplankton that act as intermediate hosts of cestodes. One hundred and ninety four copepods of the suborder Calanoida, 317 copepods of the suborder Cyclopoida and 4240 cladocerans were collected in the Juqueri reservoir, in the state of Sao Paulo, from January to August, 2003. Only Copepods Calanoida of the genus Notodiaptomus sp. Kiefer were found to be infected and contained two distinct forms of metacestodes. The metacestodes, denominated Met 1 (order Proteocephalidea) and Met 2 (order Cyclophyllidea), had the following rates of prevalence and mean intensities of infection: Met 1 - 2.06% and 64 larvae/copepod and Met 2 - 0.52% and one larvae/copepod. The positive copepods were collected at the margins of the reservoir during the day. This finding suggest that parasitism may lead to a change in the behavior of the copepods and make them more susceptible to predation in shallow water.

Síntese e atividade biológica do derivado 6-formil-oxamniquina

Schistosomiasis, an important disease in Brazil, is caused by a trematode of the genus Schistosoma, reaching millions of person in one of the most endemic region of this disease in the whole globe. The main goal of this work was to syntetize the 6-formyl-oxamniquine derivative and evaluate its biological activity. The 6-formyl-oxamniquine derivative was obtained by the oxidation of oxamniquine with MnO2, applying CH2Cl2 as solvent at room temperature for 24 hours. The obtaintion of 6-formyl-oxamniquine derivative compound was confirmed by IR spectroscopy and 13C NMR and 1H NMR, presenting similar activity when compared to the commercial oxamniquine (Mansil®).