Effects of the Epap granulovirus on its host,Epinotia aporema (Lepidoptera: Tortricidae)

The bean shoot borer, Epinotia aporema, is a major pest of soybeans in Argentina. Larvae of this pest are attacked by a granulovirus (EpapGV) that is the most important cause of sporadic epizootics in E. aporema populations. We studied the pathology of this virus in last-instar larvae using light and electron microscopy, and evaluated the effect of the disease on larval growth and development. EpapGV caused a polyorganotropic infection. No nucleocapsids were observed in the nuclei of infected cells prior to nuclear membrane disruption. Nevertheless, granulin was detected in the nucleus by immuno-gold staining, indicating that late gene expression occurred prior to nuclear membrane disruption. Establishment of the virogenic stroma led to complexes of continuous parallel convoluted membranous sheets. Nucleocapsids were enveloped in these areas to form virions, which were then occluded. Apparently as part of the cell-to-cell spread of infection, nucleocapsids were observed enclosed in large numbers within membrane-bound vesicles located between the cells and basal lamina. Larvae infected by EpapGV suffered a retardation of development and typically failed to pupate, but exhibited a weight increase greater than that of healthy E. aporema.     

TCR engagement induces proline-rich tyrosine kinase-2 (Pyk2) translocation to the T cell-APC interface independently of Pyk2 activity and in an immunoreceptor tyrosine-based activation motif-mediated fashion

The relocation of kinases in T lymphocytes during their cognate interaction with APCs is essential for lymphocyte activation. We found that the proline-rich tyrosine kinase-2 (Pyk2) is rapidly translocated to the T cell-APC contact area upon T cell-specific recognition of superantigen-pulsed APCs. Stimulation with anti-CD3-coated latex microspheres was sufficient for Pyk2 reorientation, and the coengagement of CD28 boosted Pyk2 redistribution. Nevertheless, Pyk2 translocation did not result in its recruitment to lipid rafts. Two results support that Pyk2 translocation was independent of its kinase activity. First, Lck activity was required for TCR-induced Pyk2 translocation, but not for TCR-induced Pyk2 activation. Second, a kinase-dead Pyk2 mutant was equally translocated upon TCR triggering. In addition, Lck activity alone was insufficient to induce Pyk2 reorientation and activation, requiring the presence of at least one intact immunoreceptor tyrosine-based activation motif (ITAM). Despite the dependence on functional Lck and on phosphorylated ITAM for Pyk2 translocation, the ITAM-binding tyrosine kinase zeta-associated protein 70 (ZAP-70) was not essential. All these data suggest that, by translocating to the vicinity of the immune synapse, Pyk2 could play an essential role in T cell activation and polarized secretion of cytokines.     

Triatomines involved in domestic and wild Trypanosoma cruzi transmission in Concepción, Corrientes, Argentina

An entomological and serological survey was performed in three localities of the Department of Concepción, Province of Corrientes, Argentina in 1998 and 1999, to identify triatomines species involved in domestic and wild transmission of Chagas disease. Triatomines were collected by man/hour capture in 32 houses randomly selected and 44 nearby outdoor ecotopes. Trypanosoma cruzi infection in triatomines was assessed by direct microscopic observation (400x) of feces and polymerase chain reaction. Serological techniques used for people were Indirect Hemagglutination Test and Indirect Fluorescent Test. Triatomines were collected in 28.1% of the houses and 31.8% of the wild biotopes. Triatoma infestans (Klug 1834) was exclusively found indoors and T. cruzi infected 60% of them. Triatoma sordida (St?l 1859) was mainly found in extradomestic ecotopes where trypanosome infection rate reached 12.7%. Serological study of 98 local people showed that 29.6% were seroreactive; most of their houses were closed to wild biotopes colonized by T. sordida. Results indicate that there is an active T. infestans mediated transmission of Chagas disease in this zone that yields important human prevalence and that the populations of T. sordida in wild biotopes not only sustain the wild T. cruzi cycle but also represent an actual risk for people living in the area.     

Biological variability in clinical isolates of Trichomonas vaginalis

Eighteen clinical isolates of Trichomonas vaginalis were obtained from women who attended health centers of the Government of Madrid. A total of 1,848 vaginal specimens recovered during the gynaecological examination were seeded in culture tubes containing liquid Diamond medium. Pathogenicity to mice was determined after intraperitoneal inoculation of mice by quantification of mortality and gross damage to abdominal organs. As could be expected, a broad variability was obtained, being some of the isolates more virulent than the reference strain. Regarding to metronidazole susceptibility, none resistant isolate was found but different degrees of susceptibility were determined.     

Aggregation behaviour and interspecific responses in Rhodnius prolixus stål

The response to intra- and interspecific faecal assembling signals was tested in Rhodnius prolixus. Papers impregnated with excrement of R. prolixus induced the aggregation of larvae of this species, but also of those of Triatoma infestans. However, faeces belonging to T. infestans were not able to assemble larvae of R. prolixus. On the other hand, there was no response of R. prolixus to putative chemical factors from their cuticle (footprints), in contrast to T. infestans. Results are discussed as related to the ecology of both species.     

In vivo and in vitro phosphorylation of Candida albicans 20S proteasome

In this paper we demonstrate that the Candida albicans 20S proteasome is in vivo phosphorylated and is a good in vitro substrate (S(0.5) 14nM) of homologous protein kinase CK2 (CK2). We identify alpha6/C2, alpha3/C9, and alpha5/Pup2 proteasome subunits as the main in vivo phosphorylated and in vitro CK2-phosphorylatable proteasome components. In vitro phosphorylation by homologous CK2 holoenzyme occurs only in the presence of polylysine, a characteristic that distinguishes the yeast proteasomes from mammalian proteasomes which are phosphorylated by CK2 in the absence of polycations. The major in vivo phosphate acceptor is the alpha3/C9 subunit, being phosphorylated in serine, both in vivo and in vitro. The phosphopeptides generated by endoproteinase Glu-C digestion from in vivo labeled alpha3/C9 subunit, from in vitro phosphorylation by homologous CK2 holoenzyme, and from the recombinant alpha3/C9 subunit phosphorylated by recombinant human CK2-alpha subunit are identical, suggesting that CK2 is likely responsible for in vivo phosphorylation of this subunit. Direct mutational analysis shows that serine 248 is the residue of the alpha3/C9 subunit phosphorylated by CK2. The in vitro stoichiometry of phosphorylation of the alpha6/C2 and alpha3/C9 proteasome subunits by CK2 can be estimated as 0.7-0.8 and 0.4-0.5 mol of phosphate per mole of subunit, respectively. These results are consistent with the relative abundance of the unphosphorylated and phosphorylated isoforms of these subunits present in the purified 20S proteasome preparation. Our demonstration of phosphorylation of C. albicans proteasome suggests that phosphorylation might be a general mechanism of regulation of proteasome activity.     

Paraoxonase and other coronary risk factors in a community-based cohort

Oxidative damage to circulating lipids and vascular tissues contributes to the initiation and progression of atherosclerosis. High density lipoprotein provides protection from atherosclerosis and the enzyme paraoxonase may contribute to this effect. The aim of the present study was to examine the trends in paraoxonase activity during the course of a community-directed life-style intervention, and relationships of paraoxonase activity to other coronary heart disease risk factors, in a cohort of Australian Aboriginal people.     

Canine Distemper Virus DNA Vaccination Induces Humoral and Cellular Immunity and Protects against a Lethal Intracerebral Challenge

We have studied the immune responses to the two glycoproteins of the Morbillivirus canine distemper virus (CDV) after DNA vaccination of BALB/c mice. The plasmids coding for both CDV hemagglutinin (H) and fusion protein (F) induce high levels of antibodies which persist for more than 6 months. Intramuscular inoculation of the CDV DNA induces a predominantly immunoglobulin G2a (IgG2a) response (Th1 response), whereas gene gun immunization with CDV H evokes exclusively an IgG1 response (Th2 response). In contrast, the CDV F gene elicited a mixed, IgG1 and IgG2a response. Mice vaccinated (by gene gun) with either the CDV H or F DNA showed a class I-restricted cytotoxic lymphocyte response. Immunized mice challenged intracerebrally with a lethal dose of a neurovirulent strain of CDV were protected. However, approximately 30% of the mice vaccinated with the CDV F DNA became obese in the first 2 months following the challenge. This was not correlated with the serum antibody levels.Inoculation of plasmid DNA into muscle and the subsequent expression of the encoded protein have opened up new approaches in vaccination and gene therapy (for an extensive review see reference 25). Although initial studies used intramuscular (i.m.) inoculation to deliver the DNA, other routes have been shown to be equally or more efficient in inducing immune responses, which may be related to the types of antigen-presenting cells (APCs) which are involved (9). Recent observations suggest that after i.m. inoculation, muscle cells probably act as a reservoir for the foreign antigen, while the bone marrow cells seem to act as the APCs (8, 12, 26, 27). For DNA delivery to the skin, the APCs have not yet been identified but could well include cells of dendritic origin (21). Although both intradermal and i.m. DNA inoculations induce a strong Th1 response, inoculation of DNA precipitated onto gold beads and delivered by means of a gene gun favors a Th2 response (7). Whether this is due to the targeting of different APCs has not been determined.We have been studying the possibility of using DNA vaccination to protect against canine distemper virus (CDV). CDV is a member of the genus Morbillivirus, in the Paramyxoviridae family, and although this virus primarily infects dogs, the disease has also been described in several animal species both in nature and in captivity (10, 18, 22). The currently available live attenuated vaccine efficiently protects dogs once maternal antibodies have disappeared, but it is not sufficiently attenuated for certain other animal species in which a fatal infection may ensue (6). This has led to a problem in protecting members of rare animal species living in captivity.In the present study, we have expressed the two CDV glycoproteins, the attachment protein (hemagglutinin [H]) and the fusion protein (F), from plasmids driven by a cytomegalovirus (CMV) promoter. We show that i.m. and intradermal inoculation of the CDV H-encoding plasmid induces a Th1 response, whereas gene gun inoculation of the same plasmid induces a Th2-type response. In contrast, the CDV F gene administered with the gene gun elicited a mixed Th response. Mice immunized with either of the plasmids were protected against a lethal intracerebral (i.c.) infection.     

Evidence of rodent pollination inCajophora coronata (Loasaceae)

Vegetative and floral features ofCajophora coronata (Loasaceae) suggest adaptations to pollination by rodents: (1) mammal guard represented by a covering of stinging hairs; (2) geoflory; (3) white corolla; (4) open flowers with copious low concentration nectar; (5) abundant pollen; (6) maximum pollen and nectar presentation in the afternoon hours and in the night. Palynological analysis revealed pollen loads ofC. coronata on the nostrils and whiskers of captured rodents (Graomys griseoflavus, fam.Muridae). Pollen and anther remains were also found in faeces sampled in the surroundings. Additional evidence includes rodent footprints obtained by placing smoked plates beneath the flowers, which revealed flower visitation during the night. These observations are to our knowledge the first evidence of flower visitation by rodents in South America and the first in the New World outside the range of flower bats and bat flowers.     

Chemotaxonomic features of lipophilic components inAdesmia species (Leguminosae)

Studies were performed on GC-MS to assess the lipophilic composition of sixAdesmia species representing two subgenera and three series. Normal fatty acids and hydrocarbons were mainly found, as well as acetylenic compounds, dibasic acids, cyclic hydrocarbons, high molecular weight alcohols and one sterol.