A new ascarid species in cynodont coprolite dated of 240 million years.

Cynodonts represent the transition from reptiles to mammals. They are classified as synapsids, or tetrapod animals with mammalian characteristics. We present here the finding of helminth eggs in a coprolite identified as of cynodont origin dated of nearly 240 million years. Microscopy revealed the presence of very well preserved intestinal parasite eggs. Up to now we identified an ascarid egg by morphological characteristics. Based on a previous description of the new genus Ascarites Poinar Jr and Boucot 2006 in coprolites of iguanodons from Belgium, we propose a new species, Ascarites rufferi n.sp. in cynodonts, a host that inhabited the Southern Region of Brazil in the Triassic period.

Lutz's spontaneous sedimentation technique and the paleoparasitological analysis of sambaqui (shell mound) sediments.

Parasite findings in sambaquis (shell mounds) are scarce. Although the 121 shell mound samples were previously analysed in our laboratory, we only recently obtained the first positive results. In the sambaqui of Guapi, Rio de Janeiro, Brazil, paleoparasitological analysis was performed on sediment samples collected from various archaeological layers, including the superficial layer as a control. Eggs of Acanthocephala, Ascaridoidea and Heterakoidea were found in the archaeological layers. We applied various techniques and concluded that Lutz's spontaneous sedimentation technique is effective for concentrating parasite eggs in sambaqui soil for microscopic analysis.

The C-terminal extension of Leishmania pifanoi amastigote-specific cysteine proteinase Lpcys2: a putative function in macrophage infection.

Cysteine proteinases have been implicated in many aspects of protozoan parasite pathogenesis. These hydrolases are normally found as zymogens, and some classes in trypanosomatids possess a long C-terminal extension (CTE), for which no function has been assigned. In this paper we hypothesize that the CTE domain of Lpcys2, the abundant lysosomal cysteine proteinase of Leishmania pifanoi amastigotes, is involved in host cell infection. Confirming previous reports that this peptide is highly immunogenic in Trypanosoma cruzi, we detected antibodies against CTE in sera of leishmaniasis patients. We produced a polyclonal antibody specific to Lpcys2 CTE and determined that this antibody was capable of recognizing both L. pifanoi and Leishmania amazonensis cysteine proteinases. Using this antibody, we detected a predominant localization of Lpcys2 CTE in the lysosome and flagellar pocket of cultured axenic amastigotes of both parasite species; however, its location was shifted towards the surface of the parasites during macrophage infection. We examined the role of Lpcys2 CTE in macrophage infection and found a significant reduction in the percentage of infected cells when macrophages were infected with L. pifanoi and L. amazonensis in the presence of anti-CTE antibody. This study suggests a role for leishmanial cysteine proteinases CTE at early stages of infection.

Prehistorical Pediculus humanus capitis infestation: quantitative data and low vacuum scanning microscopy.

A pre-Columbian Peruvian scalp was examined decades ago by a researcher from the Oswaldo Cruz Foundation. Professor Olympio da Fonseca Filho described nits and adult lice attached to hair shafts and commented about the origin of head lice infestations on mankind. This same scalp was sent to our laboratory and is the subject of the present paper. Analysis showed a massive infestation with nine eggs/cm2 and an impressive number of very well preserved adult lice. The infestation age was roughly estimated as nine months before death based on the distance of nits from the hair root and the medium rate of hair growth. A small traditional textile was associated with the scalp, possibly part of the funerary belongings. Other morphological aspects visualized by low-vacuum scanning electron microscopy are also presented here for adults and nits.

PREHISTORICAL Pediculus humanus capitis INFESTATION: QUANTITATIVE DATA AND LOW VACUUM SCANNING MICROSCOPY / Infestação pré-histórica por Pediculus humanus capitis: análise quantitativa e por microscopia de varredura de baixo vácuo

A pre-Columbian Peruvian scalp was examined decades ago by a researcher from the Oswaldo Cruz Foundation. Professor Olympio da Fonseca Filho described nits and adult lice attached to hair shafts and commented about the origin of head lice infestations on mankind. This same scalp was sent to our laboratory and is the subject of the present paper. Analysis showed a massive infestation with nine eggs/cm2 and an impressive number of very well preserved adult lice. The infestation age was roughly estimated as nine months before death based on the distance of nits from the hair root and the medium rate of hair growth. A small traditional textile was associated with the scalp, possibly part of the funerary belongings. Other morphological aspects visualized by low-vacuum scanning electron microscopy are also presented here for adults and nits.

Há décadas um escalpo peruano, datado do período pré-colombiano, foi examinado por um pesquisador da Fundação Oswaldo Cruz. O Professor Olympio da Fonseca Filho descreveu lêndeas e adultos fixos a fios de cabelos e fez comentários sobre a origem da infecção por piolhos na espécie humana. Este mesmo escalpo foi enviado ao nosso laboratório e é objeto deste artigo. Sua análise mostrou maciça infestação, com nove lêndeas/cm2 em impressionante número de adultos muito bem preservados. O tempo de infestação foi estimado em cerca de nove meses antes da morte, baseado na maior distância entre lêndeas e o couro cabeludo, levando em consideração taxa média de crescimento capilar de 1 cm por mês. Um pequeno pedaço de tecido tradicional peruano foi encontrado associado ao escalpo, provavelmente pertencente ao conjunto de peças usado no ritual funerário. Aqui, apresentamos alguns aspectos morfológicos de adultos e lêndeas vizualizados por microscopia eletrônica de varredura de baixo vácuo.

Cellular signaling during the macrophage invasion by Trypanosoma cruzi.

We have reported that protein tyrosine kinases play an important role in the invasion of Trypanosoma cruzi into primary resident macrophages. In the present study we carry out immunofluorescence assays, using monoclonal anti-phosphotyrosine antibodies, to reveal an accumulation of tyrosine-phosphorylated residues at the site of parasite association with the macrophage surface, colocalizing with host cell F-actin-rich domains. SDS-PAGE analysis of macrophage cell line IC-21 tyrosine phosphoproteins, labeled with [(35)S] L-methionine, revealed several peptides with increased levels of phosphorylation upon interaction with the parasite. Among them, were detected bands of 140, 120, 112, 94, 73, 67, and 56 kDa that match the molecular weights of proteins described as being tyrosine phosphorylated during events that lead to actin assembly in mononuclear phagocytes. The pretreatment of IC-21 macrophages with the tyrosine kinase inhibitor tyrphostin 23 inhibited trypomastigote uptake showing that tyrosine phosphorylation is important for the parasite penetration in this particular cell line. Immunofluorescence microscopy, using antibodies against p85, the regulatory subunit of phosphatidylinositol 3-kinase (PI 3-kinase), placed this enzyme also in the same sites, in accordance to what is reported for phagocytosis. We suggest that once the components of T. cruzi trypomastigotes surface are recognized by macrophage receptors, they trigger the activation of a tyrosine phosphorylation cascade, PI 3-kinase recruitment, and assembly of actin filaments at the site of initial cell-to-cell contact, resembling the events described during phagocytosis. These achievements support the model for a phagocytic-like actin-dependent invasion mechanism for T. cruzi trypomastigotes into macrophages.