Activity of "reversed" diamidines against Trypanosoma cruzi "in vitro"

Chagas' disease is an important parasitic illness caused by the flagellated protozoan Trypanosoma cruzi. The disease affects nearly 17 million individuals in endemic areas of Latin America and the current chemotherapy is quite unsatisfactory based on nitroheterocyclic agents (nifurtimox and benznidazol). The need for new compounds with different modes of action is clear. Due to the broad-spectrum antimicrobial activity of the aromatic dicationic compounds, this study focused on the activity of four such diamidines (DB811, DB889, DB786, DB702) and a closely related diguanidine (DB711) against bloodstream trypomastigotes as well as intracellular amastigotes of T. cruzi in vitro. Additional studies were also conducted to access the toxicity of the compounds against mammalian cells in vitro. Our data show that the four diamidines compounds presented early and high anti-parasitic activity (IC50 in low-micromolecular range) exhibiting trypanocidal dose-dependent effects against both trypomastigote and amastigote forms of T. cruzi 2h after drug treatment. Most of the diamidines compounds (except the DB702) exerted high anti-parasitic activity and low toxicity to the mammalian cells. Our results show the activity of reversed diamidines against T. cruzi and suggested that the compounds merit in vivo studies.     

Mammalian target of rapamycin complex 1 is involved in differentiation of regenerating myofibers in vivo

This work was undertaken to provide further insight into the role of mammalian target of rapamycin complex 1 (mTORC1) in skeletal muscle regeneration, focusing on myofiber size recovery. Rats were treated or not with rapamycin, an mTORC1 inhibitor. Soleus muscles were then subjected to cryolesion and analyzed 1, 10, and 21 days later. A decrease in soleus myofiber cross-section area on post-cryolesion days 10 and 21 was accentuated by rapamycin, which was also effective in reducing protein synthesis in these freeze-injured muscles. The incidence of proliferating satellite cells during regeneration was unaltered by rapamycin, although immunolabeling for neonatal myosin heavy chain (MHC) was weaker in cryolesion+rapamycin muscles than in cryolesion-only muscles. In addition, the decline in tetanic contraction of freeze-injured muscles was accentuated by rapamycin. This study indicates that mTORC1 plays a key role in the recovery of muscle mass and the differentiation of regenerating myofibers, independently of necrosis and satellite cell proliferation mechanisms.     

Acute lung injury in experimental pancreatitis in rats: pulmonary protective effects of crotapotin and N-acetylcysteine

Respiratory complications are major factors contributing to death in acute pancreatitis. However, the mechanisms of these pulmonary complications are not completely elucidated. We studied the effects of pretreatment with purified crotapotin (a phospholipase A2 inhibitor), N-acetylcysteine (a reactive oxygen species inhibitor), and a combination of both on the pulmonary mechanical and morphometric changes secondary to severe acute necrohemorrhagic pancreatitis in Wistar rats. A total of 69 male Wistar rats were studied. Pancreatitis was induced by infusion of 0.5 mL of a 4% solution of sodium taurocholate into the biliopancreatic duct. Crotapotin, N-acetylcysteine, or a combination of both was given intraperitoneally 30 min before inducing pancreatitis. Data were compared with data from sham-operated animals with or without those pretreatments. The severity of pancreatic and pulmonary injuries was evaluated 4 h after inducing pancreatitis by morphometric and pulmonary mechanical studies. N-acetylcysteine prevented the development of alveolar edema, alveolar distention, and collapse. Crotapotin prevented alveolar distention and collapse, and pulmonary dynamic elastance increase. When used in combination, crotapotin and N-acetylcysteine prevented both pulmonary morphological and mechanical changes induced by acute pancreatitis, suggesting an increase in protective effect when these drugs are used together compared with individual effects. However, the severity of pancreatic necrosis and the increase in polymorphonuclear cells in alveolar septa induced by pancreatitis were not reduced by previous administration of crotapotin, N-acetylcysteine, or both. These results suggest that the protective effects of these drugs are probably due to an extra-pancreatic action in the circulation, or even directly in the lung.     

Formation of human myocardium in the rat heart from human embryonic stem cells

Human embryonic stem cells (hESCs) offer the opportunity to replenish cells lost in the postinfarct heart. We explored whether human myocardium could be generated in rat hearts by injecting differentiated cardiac-enriched hESC progeny into the left ventricular wall of athymic rats. Although initial grafts were predominantly epithelial, noncardiac elements were lost over time, and grafts consisted predominantly of cardiomyocytes by 4 weeks. No teratomatous elements were observed. Engrafted cardiomyocytes were glycogen-rich and expressed expected cardiac markers including beta-myosin heavy chain, myosin light chain 2v, and atrial natriuretic factor. Heat-shock treatment improved graft size approximately threefold. The cardiac implants exhibited substantial angiogenesis, both recipient and graft derived. Importantly, there was greater proliferation in human cardiomyocytes than previously seen in rodent-derived cardiomyocytes: 14.4% of graft cardiomyocytes expressed the proliferation marker Ki-67, and 2.7% incorporated the thymidine analog BrdU 4 weeks after transplantation. This proliferation was associated with a sevenfold increase in graft size over the 4-week interval. Thus, hESCs can form human myocardium in the rat heart, permitting studies of human myocardial development and physiology and supporting the feasibility of their use in myocardial repair.     

Molecular epidemiology of dengue type 3 virus in Brazil and Paraguay, 2002-2004.

We studied the molecular epidemiology of dengue virus type 3 (DENV-3) in Brazil and Paraguay by analyzing the 5' and 3' untranslated regions (5' and 3'UTRs) and the E protein gene of viruses isolated between 2002 and 2004. Both 5' and 3'UTRs were highly conserved. However, the 3'UTR of two isolates from Brazil contained eight nucleotide deletions compared with the remaining 26 viruses. Phylogenetic analyses suggested that DENV-3 was introduced into Brazil from the Caribbean Islands at least twice and into Paraguay from Brazil at least three times.