María Leonor Caldas

Cytochemical localisation of calcium ATPase activity during the erythrocytic cell cycle of Plasmodium falciparum

Using a cytochemical technique, we evaluated the levels of Ca(2+)-ATPase activity in the plasmatic and in the parasitophorous vacuole membrane through the different developmental stages of the Plasmodium falciparum parasitised erythrocyte. We found that the activity is detectable and remains unaltered in the plasma membrane throughout the 48 h cell cycle. However, in the parasitophorous membrane, although the activity was very similar to that measured in the plasma membrane of the young stages (younger than 20-h-old parasites), it diminished gradually with maturation and in schizonts it was almost undetectable. These data suggest that the plasma membrane Ca(2+)-ATPase is important in the maintenance of a low erythrocyte cytoplasmic Ca(2+) concentration, and that in addition it could be a way to supply the vital cation to the parasite at the beginning of the infection, when other transport mechanisms have not yet developed.

Effect of an Oxadiazoline and a Lignan on Mycolic Acid Biosynthesis and Ultrastructural Changes of Mycobacterium tuberculosis.

Tuberculosis (TB) is an important disease that causes thousands of deaths around the world. Resistance against antitubercular available drugs has been reported; so, research on new effective antimycobacterial molecules is needed. Antimycobacterial activity of three lignans and two synthetic hydrazones was assessed against Mycobacterium tuberculosis H37Rv by antimycobacterial microdilution assay (TEMA). An oxadiazoline (AC451) and a lignan (ethoxycubebin) were the most active compounds (MIC 6.09 and 62.4 μM, resp.). Several changes in mycolic acid profile of treated bacteria were detected with both compounds by mass spectrometry analysis. Additionally, the level of reduction of mycolic acids in ethoxycubebin treatment was correlated to disruption in bacterial morphology.

Cellular location of KMP-11 protein in Trypanosoma rangeli.

We describe the localization of the KMP-11 protein in the Trypanosoma rangeli parasite determined by immunoelectron microscopy using a monoclonal antibody generated against the Trypanosoma cruzi KMP-11 protein. The data reported herein show that the T. rangeli KMP-11 protein is mainly accumulated in the parasite cytoplasm, the coat, the flagellum, and the flagellar pocket. The high degree of sequence homology between the KMP-11 proteins from both parasites suggests that the KMP-11 protein from T. rangeli, like that of T. cruzi, could also be associated with the parasite cytoskeleton.

Cambios ultrastructurales en núcleos de células C6/36 infectadas con virus dengue de tipo 2

INTRODUCTION Dengue virus replication has been considered mainly cytoplasmic, however, studies indicate that some flaviviruses may use the intranuclear pathway as part of the machinery that the virus uses to increase infection capacity in the host cell. This paper describes alterations at nuclear level in the cell infected with dengue, which are likely involved in the virus replication processes. OBJECTIVE This paper addresses the ultrastructural observations of C6/36 cells of the Aedes albopictus mosquito infected with dengue virus type 2. MATERIALS AND METHODS C6/36 cells were infected in culture medium with the serum of a patient positively diagnosed for dengue 2. Subsequently, the cells were incubated for 10 days and the cytopathic effect was assessed. The cells were processed for immunofluorescence assays and transmission electron microscopy. RESULTS The immunofluorescence assays confirmed the presence of viral protein E associated with cellular syncytia in the culture. In the ultrastructural study, the infected cells showed vesicular-tubular structures and dilated cisterns of the endoplasmic reticulum at the cytoplasmic level. Viral particles were found exclusively in cytoplasm localized within the vacuoles. Nuclei of cellular syncytia showed membrane structures arranged in a circular shape and, in some cases, these syncytia displayed lysis; in no case viral particles were observed at the nuclear level. CONCLUSIONS The ultrastructural alterations of nuclei in cells infected with the dengue virus using electron microscopy techniques had not been reported before, as far as we know. It is likely that such modifications are associated with replicative processes at an intranuclear level as an alternate replication mechanism.

Glucógeno hepático en dengue severo: análisis histopatológico

Background: Dengue virus affects various organs, but the liver is the main target of damage and where the most severe damage can occur. There are few studies on the histological changes in the liver during dengue infection. Aims: To analyze the histopathological post-mortem alterations in livers from patients with severe dengue.

Morphological changes in lung tissue of victims associated with the 2009 A H1N1/v09 influenza pandemic in Colombia

INTRODUCTION Influenza is an acute respiratory infection that may be seasonal or pandemic. In 2009 The World Health Organization (WHO) declared an influenza pandemia; 3,876 cases and 239 deaths were reported in Colombia. OBJECTIVE The morphological changes in lung tissues associated with virus infection H1N1/v09 were described from autopsied victims. Materials and methods. Seventy-five cases were diagnosed by RT-PCR for influenza A H1N1/v09, of which the lungs of 20 were selected for morphological study by light microscopy, optical microscopy, high-resolution transmission electron microscopy and immunohistochemistry. RESULTS Of the 75 cases, 83% had viral pneumonitis and 17% alveolitis. Complications included intra-alveolar hemorrhage (66%), edema (89%), diffuse alveolar damage (2%), and bacterial co-infection (32%). Morphological changes were as follows: destruction of the alveolar epithelium and interstitium, edema, macrophages with vacuolated cytoplasm,and infiltration of polymorphonuclear leukocytes in the alveolar lumen and interstitium, vacuolization cytoplasmic type I pneumocytes and electronedense bodies in cellular debris in the alveolar lumen, and immunoreactivity of viral antigens in bronchiolar epithelial cells and alveolar infiltrate. CONCLUSION The low percentage of bacterial co-infection observed in these cases was a prominent feature, and suggested that the fatal result was probably not associated with secondary bacterial disease (Indicated by previous reports). The tissue lesions were attributed to tissue damage due to viral lesion, as well as the cellular and humoral inflammatory response associated with infiltration by polymorphonucleocytes and macrophages in the interstitium and alveolar lumen.

Matrices de inclusión para obtención de cortes de tejidos en vibrátomo en estudios citoquímicos

Para estudios citoquimicos en diferentes muestras de tejidos, de tamanos pequenos, se hace necesaria la utilizacion de una matriz inerte que genere soporte y estabilidad al tejido en el momento de obtener cortes de 50 pm en vibratomo. Por esta razon se ensayaron cinco matrices: agar granulado, agar purificado, agar-agar, agarosa y gelatina, a diferentes concentraciones.Tanto con el agar-agar como con la agarosa se obtuvieron los mejores resultados, puesto que estos ofrecen mayor estabilidad al tejido, fluyen facilmente al servirse y su textura es muy homogenea, requisitos necesarios para la obtencion de cortes precisos, utilizables en citoquimica, inmunocitoquimica y trazados histoquimicos.