Luiz Augusto Corrêa Passos

Transmission of Chagas Disease (American Trypanosomiasis) by Food

In April 2009, the centenary of the discovery of the American trypanosomiasis, or Chagas disease, was celebrated. A hundred years after the discovery, little has been invested in diagnostics and treatment because the disease affects mainly poor people in developing countries. However, some changes in the epidemiology of the disease are of great importance today. Chagas disease transmitted through food is a public health concern in all areas where there is a reservoir of Trypanosoma cruzi in wild animals (e.g., mammals and marsupials) and/or where infected triatomine bugs are in contact with human food source items (especially fruits and vegetables). Recently, several outbreaks of illness related to the ingestion of food contaminated with T. cruzi have been recorded in Brazil, Colombia, and Venezuela.

Increased Cavernosal Relaxations in Sickle Cell Mice Priapism are Associated with Alterations in the NO‐cGMP Signaling Pathway

INTRODUCTION Priapism is defined as prolonged and persistent penile erection, unassociated with sexual interest or stimulation, and is one of the many serious complications associated with sickle cell disease (SCD). AIM The aim of this study was to evaluate the role of the NO-cGMP signaling pathway in priapism in Berkeley murine model of SCD (SS). METHODS SS mice and C57BL/6 mice (control) penile tissues were removed and the erectile tissue within the corpus cavernosum (CC) was surgically dissected free. The strips were mounted in 10 mL organ baths containing Krebs solution at 37 degrees C (95% O(2), 5% CO(2), pH 7.4), and vertically suspended between two metal hooks. MAIN OUTCOME MEASURES Cumulative concentration-response curves were constructed for acetylcholine (ACh; endothelium-dependent responses), sodium nitroprusside (SNP; endothelium-independent relaxations) and BAY 41-2272 (a potent activator of NO-independent site of soluble guanylate cyclase) in CC precontracted with phenylephrine. Cavernosal responses induced by frequency-dependent electrical field stimulation (EFS) were also carried out to evaluate the nitrergic cavernosal relaxations. RESULTS In SS mice, ACh-induced cavernosal relaxations were leftward shifted by 2.6-fold (P < 0.01) that was accompanied by increases in the maximal responses (78 +/- 5% and 60 +/- 3% in SS and C57B6/6J mice, respectively). Similarly, SNP- and BAY 41-2272-induced CC relaxations were leftward shifted by approximately 3.3- and 2.2-fold (P < 0.01) in SS mice, respectively. A significant increase in maximal responses to SNP and BAY 41-2272 in SS mice was also observed (113 +/- 6% and 124 +/- 5%, respectively) compared with C57B6/6J mice (83 +/- 4% and 99 +/- 2%, respectively). The EFS-induced cavernosal relaxations were also significantly higher SS mice. CONCLUSION These results showed that SS mice exhibit amplified corpus carvenosum relaxation response mediated by NO-cGMP signaling pathway. Intervention in this signaling pathway may be a potential therapeutic target to treat SCD priapism.

Parasite survey in mouse and rat colonies of Brazilian laboratory animal houses kept under differents sanitary barrier conditions

A parasitological study was undertaken to determine the health status of 15 mouse and 10 rat colonies bred in 18 Brazilian laboratory animal houses maintained under different sanitary barrier conditions which supply animals for teaching, research purposes and manufacture of biological products for medical or veterinary use. Parasitological methods were used for diagnosis of mites, lices, helminthes and protozoan parasites. A questionnaire was answered by institutions with the intention to obtain information about the existence of barriers against infections and of regular sanitary monitoring program of their colonies. The questionnaire data show that the majority of the animal houses investigated do not possess an efficient sanitary barrier system able to keep animals under controlled health sanitary conditions. Ecto and endoparasite infections are widespread in the colonies and multiple infections were common in animals from most facilities investigated. The prevalences of parasites detected among the mouse and rat colonies of the laboratory animal houses investigated were: Myocoptes musculinus (46.6%), Myobia musculi (26.6%), Radfordia ensifera (13.3%), Syphacia obvelata (86.6%), Aspiculuris tetraptera (60.0%), Hymenolepis nana (53.3%), Spironucleus muris (80.0%), Tritrichomonas muris (80.0%), Giardia muris (66.0%), Entamoeba muris (20.0%), Eimeria sp. (13.3%), Hexamastix muris (26.6%), Poliplax spinulosa (30.0%), Poliplax serrata (10.0%), Radfordia ensifera (30.0%), Syphacia muris (80.0%), Hymenolepis nana (40.0%), Trichosomoides crassicauda (55.5%), Spironucleus muris (90.0%), Tritrichomonas muris (80.0%), Giardia muris (60.0%), Entamoeba muris (80.0%), Eimeria sp. (60.0%) and Hexamastix muris (60.0%).

The Contribution of Nicotinamide Nucleotide Transhydrogenase to Peroxide Detoxification Is Dependent on the Respiratory State and Counterbalanced by Other Sources of NADPH in Liver Mitochondria.

The forward reaction of nicotinamide nucleotide transhydrogenase (NNT) reduces NADP+ at the expense of NADH oxidation and H+ movement down the electrochemical potential across the inner mitochondrial membrane, establishing an NADPH/NADP+ ratio severalfold higher than the NADH/NAD+ ratio in the matrix. In turn, NADPH drives processes, such as peroxide detoxification and reductive biosynthesis. In this study, we generated a congenic mouse model carrying a mutated NntC57BL/6J allele from the C57BL/6J substrain. Suspensions of isolated mitochondria from Nnt+/+, Nnt+/−, and Nnt−/− mouse liver were biochemically evaluated and challenged with exogenous peroxide under different respiratory states. The respiratory substrates were also varied, and the participation of concurrent NADPH sources (i.e. isocitrate dehydrogenase-2, malic enzymes, and glutamate dehydrogenase) was assessed. The principal findings include the following: Nnt+/− and Nnt−/− exhibit ∼50% and absent NNT activity, respectively, but the activities of concurrent NADPH sources are unchanged. The lack of NNT activity in Nnt−/− mice impairs peroxide metabolism in intact mitochondria. The contribution of NNT to peroxide metabolism is decreased during ADP phosphorylation compared with the non-phosphorylating state; however, it is accompanied by increased contributions of concurrent NADPH sources, especially glutamate dehydrogenase. NNT makes a major contribution to peroxide metabolism during the blockage of mitochondrial electron transport. Interestingly, peroxide metabolism in the Nnt+/− mitochondria matched that in the Nnt+/+ mitochondria. Overall, this study demonstrates that the respiratory state and/or substrates that sustain energy metabolism markedly influence the relative contribution of NNT (i.e. varies between nearly 0 and 100%) to NADPH-dependent mitochondrial peroxide metabolism.

Survival In Vitro and Virulence of Trypanosoma cruzi in Acai Pulp in Experimental Acute Chagas Disease

Chagas disease is a parasitic infection with high socioeconomic impact throughout Latin America. Although this severe, incurable disease can be transmitted by several routes, oral transmission is currently the most important route in the Amazon Basin. Açaí pulp has nutritional properties and is popular throughout Brazil and abroad. However, this pulp has been associated with microepidemics of acute Chagas disease (ACD) in northern Brazil, where açaí fruit is the main food supplement. In this study, we examined the in vitro survival and in vivo virulence of Trypanosoma cruzi Y strain in açaí pulp. Aliquots of in natura açaí pulp produced in Belém city in the northern Brazilian state of Pará were mixed with 10⁵ trypomastigotes. The samples were incubated at room temperature or at 4 or -20°C for various periods, and the parasites were isolated by forced sieving. The resulting eluates were examined by microscopy, and the trypomastigotes were administered intraperitoneally, orally, or by gavage to immunodeficient mice (C.B-17-Prkdc(scid)/PasUnib) that had been pretreated with antibiotics. Parasitemia was quantified by the Brener method, and mortality was recorded daily. All routes of administration resulted in ACD. A 5-day delay in the onset of parasitemia occurred with oral administration. The survival and virulence of the parasites were unaffected by prior incubation at room temperature for 24 h, at 4°C for 144 h, and at -20°C for 26 h. These results indicate that T. cruzi can survive and retain its virulence in açaí pulp under various conditions and that cooling and freezing are not suitable methods for preventing foodborne ACD.

Virulence of Trypanosoma cruzi in Açai (Euterpe oleraceae Martius) Pulp following Mild Heat Treatment

Outbreaks of acute Chagas disease (ACD) in northern Brazil can be caused by the ingestion of unprocessed açai pulp contaminated with Trypanosoma cruzi . The aim of this study was to determine the minimum thermal process required to inactivate T. cruzi in açai pulp. Trypomastigotes (100,000) of T. cruzi Y strain were added to 0.15 M NaCl or açai pulp and continuously mixed while being heat treated at 37 to 49°C for up to 1 h. When necessary, parasites were separated from açai pulp by forced sieving. Inocula were administrated intraperitoneally in inbred immunodeficient C.B-17-Prkdcscid/Pas Unib mice, and the recipients were monitored for parasitemia and mortality. Mice received prophylactic antibiotic therapy by using cephalexin to prevent bacterial infection from the açai pulp. T. cruzi retained its virulence in 0.15 M NaCl and açai pulp at 44 ± 0.1°C for 10 min and at 43 ± 0.1°C for 20 min, respectively, causing ACD and death in mice up to 24 days after infection. Incubation of açai pulp inoculum above 43°C for 20 min neutralized T. cruzi virulence, thereby preventing ACD and death in murine recipients. The heating of açai pulp above 43°C for 20 min is a practical and effective measure to prevent foodborne ACD caused by T. cruzi .

High levels of human gamma-globin are expressed in adult mice carrying a transgene of the Brazilian type of hereditary persistence of fetal hemoglobin ((A)gamma -195).

Hereditary persistence of fetal hemoglobin (HPFH) is characterized by increased levels of Hb F during adult life. Nondeletional forms of HPFH are characterized by single base mutations in the Aγ and Gγ promoters, resulting in an increase of Hb F ranging from 3 to 20% in heterozygotes. Many point mutations in this region have been described, including the Aγ −195 (C>G) mutation that causes the Brazilian type of HPFH (HPFH-B). To better understand this mechanism, we have developed HPFH-B transgenic mice. mRNA levels of human γ-globin of −195 transgenic mice were clearly higher when compared with control transgenic mice bearing a wild type sequence of the γ promoter. Thus, our data indicate that the −195 mutation is the unique cause of elevation of Hb F in Brazilian HPFH. These results could provide us with an opportunity to study the modifying effects of the Hb F in the phenotype of sickle cell disease and β-thalassemia (β-thal).

Uso de monossulfeto de tetraetiltiuram no tratamento de ectoparasitas em camundongos

Casais de camundongos isogenicos das racas CBA, BALB/c e machos e femeas de Swiss heterogeneticos criados no sistema convencional, que se encontravam infectados em diferentes graus por Myobia musculi e Myocoptes musculinus, foram submetidos a quatro esquemas de tratamento com Tetmosol. A incidencia de ectoparasitas na raca C57BL/10 foi apenas 1,2%. Todos os animais da raca CBA e Swiss encontravam-se parasitados, enquanto que a infestacao do BALB/c foi de 50 a 90%. Os melhores resultados dos 4 esquemas de tratamento testados foram observados utilizando-se Tetmosol na concentracao 2,5% em banhos seriados com intervalos alternados de 3 e 4 dias, num periodo de 21 dias. A incidencia de ectoparasitas adultos em CBA, BALB/c e Swiss apos o tratamento foi de respectivamente 0%, 40% e 0%, enquanto que para ovos viaveis os respectivos resultados foram de 20%, 50% e 0%. Apos a introducao de matrizes tratadas num sistema de criacao sob barreiras, a colonia tem sido examinada sistematicamente ha 24 meses e encontra-se isenta de ectoparasitas.Colonies of CBA, C57BL/10, BALB/c and Swiss mice housed in conventional systems were found to be infected to different degrees with both Myobia musculi and Myocoptes musculinus. Groups of ten different sister-brother pairs of the isogenic mice and groups of 25 mice separated according to were infected by the parasite and then submitted to 4 different schemes of treatment with Tetmosol. C57BL/10 presented a very low incidence of parasites (1.2%). Every pair of CBA, and every Swiss mice examined, were found to be infected while only 50-90% of the pairs of BALB/c mice were infected. The best results of four different schemes were obtained by dipping the animals in 2.5% Tetmosol, at 3-4 days intervals for 21 days. The incidence of adult parasite forms in CBA, BALB/c and Swiss mice were found to be 0%, 40% and 0%, respectively, while in viable eggs it was found to be 20%, 50% and 0%, respectively.