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Apoptosis in Leishmania species & its relevance to disease pathogenesis.

Apoptosis is a morphologically distinct form of cell death necessary for embryogenesis, tissue homeostasis and disease control in metazoans. Earlier, it was thought that apoptosis is the prerogative of multicellular organisms. However, it is now evident that unicellular organisms are also capable of undergoing apoptosis. In the context of Leishmania spp., a unicellular eukaryote responsible for causing leishmaniasis, the process of apoptosis is important for successful survival. The flagellated promastigote form of the parasite resides in the midgut of the insect vector, the female sandfly and at this niche; the cell fittest to survive to pass onto the pharynx of the fly is selected by eliminating unfit cells through apoptosis. Within the mammalian host, inside the macrophage, apoptosis is necessary to regulate cell numbers and to minimize immune reactions. With most apoptosis inducing stimuli, L. donovani shows typical features of apoptotic death like cell shrinkage, nuclear condensation and DNA fragmentation. Agents capable of precipitating apoptosis in this parasite include anti-leishmanial drugs like antimony, amphotericin B, pentamidine and miltefosine. Other agents like heat shock, treatment with staurosporine, knocking out centrin gene also precipitate apoptosis of the parasites. A pivotal role in cellular apoptosis is played by the single mitochondrion of Leishmania spp., where a fall or increase in mitochondrial potential leads to cell death by apoptosis. Ca2+ appears to be a vital ion involved in Leishmania apoptosis and partial inhibition of cytosolic Ca2+ increase achieved by chelating extracellular or intracellular Ca2+ during oxidative stress results in significant rescue of the fall of the mitochondrial membrane potential and consequently apoptosis. Elucidation of the molecular events linked to apoptotic death of Leishmania spp. might help define a more comprehensive view of the cell death machinery in terms of evolutionary origin and identify new target molecules for chemotherapeutic drug development and therapeutic intervention.

Animals↗

Virulent and attenuated lines of Leishmania major: DNA karyotypes and differences in metalloproteinase GP63.

The Leishmania metalloproteinase GP63 has been reported to play important roles mainly in resistance of promastigotes to complement-mediated lysis and in interaction with macrophage receptors. On the other hand, its function in insect vectors is still unclear. We compared the structure and dosage of gp63 genes and the activity of GP63 in Leishmania major Yakimoff et Schokhor strains and lines differing in virulence for mice and ability to develop in sand flies. The results demonstrate considerable variability in amount and proteolytical activity of GP63 among L. major strains although genomic changes in the gp63 locus were not found. Attenuated LV561/AV line showed low amount and low enzymatic activity of GP63. Serial passages of attenuated parasites through either Phlebotomus duboscqi Neveu-Lemaire or through mice led to a recovery of GP63 proteolytical activity to the level present in virulent LV561/V line. Overexpression of GP63 was found in two L major strains (L119, Neal) with defective lipophosphoglycan (LPG); both these strains were capable to cause mice infection but unable to survive and multiply in sand flies. Differences were found also in karyotypes and in amount of minichromosomes amplified in some lines of the LV561 strain. The results suggest that parasite virulence is not simply correlated with the activity of GP63; however, this enzyme plays a significant role in association with other surface molecules, especially LPG. Overexpression of GP63 can compensate LPG defect in the vertebrate host but in sand flies both molecules fulfil quite different functions and the defect in LPG is lethal for the parasite. On the other hand, linear minichromosomes of about 200 kb found in some lines of the LV561 strain are associated with development in vitro and in the vector but they are not essential for the infection of the vertebrate host.

Animals↗

The low seroprevalence of tick-transmitted agents of disease in dogs from southern Ontario and Quebec.

Infectious diseases caused by pathogens transmitted by ticks and other insect vectors are an important cause of morbidity and mortality in both dogs and humans throughout North America. The purpose of this study was to determine the seroprevalence of selected vector-transmitted pathogens in southern Ontario and Quebec. Samples submitted to the Vector Borne Disease Diagnostic Laboratory (VBDDL) at the North Carolina State University College of Veterinary Medicine were evaluated for antibodies to Ehrlichia canis, Anaplasma phagocytophilum, Babesia canis, Bartonella henselae, Borrelia burgdorferi, Bartonella vinsonii subspecies berkhoffii, and Rickettsia rickettsii. Information regarding breed and the city or province from which the sample originated was recorded; however, travel history was unknown for the majority of dogs. Overall seroprevalence to these tick-borne pathogens in southern Ontario and Quebec is low compared with most regions of the United States, suggesting that veterinarians in this region of Canada should pursue diagnostic evidence of infection in dogs with a travel history or prior residence in areas endemic for exposure to tick-borne infections.

Animals↗

Antibodies from Chagas patients serum bind to the gut epithelial cell surface of Triatoma infestans.

Serum of chagasic patients with a high title of specific antibodies against T. cruzi antigens, binds epitopes in the midgut and hindgut of the insect vector Triatoma infestans free of parasites. These antigens were recognized at ultrastructural level by immunocytochemistry with serum of chagasic patients as first antibody and protein "A"-gold as marker. Controls with normal human serum were negative. The positive reaction occurs principally in the apical epithelial portion (microvilli and peritrophic membrane) of the midgut and in the cuticular layer and adjacent cytoplasm of the hindgut. In vectors infected with trypanosomes, the antigen-antibody reaction occurs similarly in the epithelium and also in the trypanosomes present in the lumen. These results suggest that there are antigenic factors in the vector gut that are incorporated by the parasite and are recognized by the immunitary system of the human patient with production of specific antibodies.

Animals↗

Temporal appearance, geographic distribution, and species of origin of bluetongue virus serotypes in the United States.

Beginning in 1973, all available laboratory and field strains of bluetongue virus (BTV) from the United States were serotyped. Of the viral strains serotyped, 27 were collected from 1953 through 1972; 173 were collected from 1973 through 1977. Although 20 BTV serotypes have been found worldwide, only BTV serotypes 10, 11, 13, and 17 have been found in the United States. Since 1973, serotypes 11 and 17 have been the prevalent serotypes. Samples were collected over a 24-year period in the United States and represent a wide geographic area and diverse host sources (sheep, cattle, wild ruminants, and insect vectors). The collection was not a statistical sampling.

Animals↗

[Phlebotomus in Spain].

The importance of leishmaniases in human medicine, on a global as well as a national level, is a matter that offers no doubts. The problems caused by these illnesses are what cause increased attention to the vector insects: the phlebotomos. This article offers an overview of the most significant aspects of the species to be found in Spain. Apart from mentioning the highlights of the studies which have been carried out in our country concerning these insects, an analysis is also made of the distribution throughout the entire country, giving a map which shows the species captured in each province. Mention is also made of the morphological characteristics of greatest taxonomic importance, including 3 illustrations on this subject, and finally, the article gives the most important data on its biology, be it in the pre-imaginal stages (eggs, larva, pupa), or in the adults.

Animals↗

Endocytosis by African trypanosomes. II. Occurrence in different life-cycle stages and intracellular sorting.

Horseradish peroxidase (HRP) and colloidal gold-labeled proteins enter many of the endocytic organelles of bloodstream forms of Trypanosoma brucei and T. congolense. However, the colloidal gold markers were excluded from substantial parts of the pathway that contained HRP. Morphometric studies revealed that HRP entered organelles that accounted for approximately 5% of the total cell volume while transferrin-gold entered organelles that comprised approximately 2% of the total cell volume. In addition, large colloidal gold particles were excluded from organelles that contained smaller gold particles. Antibodies, raised against the variable surface glycoprotein, when applied to thawed cryosections were found to label structures from which endocytosed colloidal gold coupled to bovine serum albumin (BSA) was excluded. Endocytosis was shown to occur in two in vitro propagated forms of trypanosomes, similar to those found in the insect vector (Glossina spp.). The mammal-infective metacyclic forms were similar to bloodstream forms in that they endocytosed HRP and colloidal gold markers but excluded colloidal gold from approximately 3% of the endocytic organelles. Estimation of the flagellar pocket volumes of bloodstream form T. brucei showed that this organelle occupied 0.5% to 1.4% of the total cell volume. The flagellar pocket volume of T. congolense varied between life-cycle stages, with a fractional volume of 4.4% for bloodstream forms, 2.3% for metacyclic forms and 1.4% for procyclic forms. Endocytosis of HRP, but not of protein-gold markers, occurred in procyclic (uncoated) forms. Endocytosis by procyclic forms has heretofore not been reported.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Antibody response to experimental Trypanosoma rangeli infection and its implications for immunodiagnosis of South American trypanosomiasis.

Differential immunodiagnosis of T. rangeli and T. cruzi infections in man poses a particular problem, not only because these parasites share antigenic determinants, as detected by immunofluorescence, but also because they have a similar geographical distribution, the same host range and often identical insect vectors. We show here that whereas mouse anti-T. rangeli sera have significant cross reactivity with T. cruzi by immunofluorescence, they are entirely specific when tested by ELISA, using apparently similar antigen preparations. Immunoprecipitation analysis detected relatively little cross-reactivity between heterologous antisera and parasite combinations. Intriguingly, immunization with T. rangeli epimastigotes was much more powerful than similar immunization with trypomastigotes, and the majority of the antibody was directed against a single polypeptide of apparent Mr 73kDa.

Animals↗

Developmental regulation of trypanosome mitochondrial gene expression.

Mitochondrial respiratory activities in the protozoan parasite Trypanosoma brucei are developmentally regulated. The trypanosomes in the mammalian bloodstream derive ATP entirely from glycolysis. The trypanosomes found in the midgut of the insect vector or in culture at 26 degrees C have fully functional mitochondria with cytochrome-mediated respiration. In this paper, we show that the steady state levels of the 9 S and 12 S mitochondrial ribosomal RNAs (rRNAs) are 30-fold lower in an early developmental stage in the mammal, the slender forms, relative to the levels in the stumpy trypanosomes, a later developmental stage in the mammalian infection. Transcripts from three other mitochondrial genes, cytochrome b and subunits I and II of cytochrome oxidase, are undetectable in the slender trypanosomes and increase in the stumpy trypanosomes to levels approaching those in trypanosomes from 26 degrees C cultures. Transcription of other mitochondrial genes, including NADH-dehydrogenase subunit 5, is unregulated during trypanosome development. These results show that the level of some mitochondrial transcripts is developmentally regulated in bloodstream trypanosomes and suggest that the stumpy bloodstream trypanosomes accumulate mitochondrial transcripts prior to development of a functional mitochondrion. These results also show that the developmental activation of mitochondrial activities at 26 degrees C is not controlled at the level of mitochondrial transcription.

Animals↗

HIV infection through normal heterosexual contact in Uganda.

One hundred and fourteen household contacts of 23 patients with AIDS or suspected AIDS were examined for evidence of infection with HIV. In total 12 contacts had antibodies to the virus by a competitive enzyme-linked immunosorbent assay (ELISA), confirmed by Western blot. Ten of these seropositive contacts were the sexual partners of the index cases. Overall 71% of sexual partners (10:14) were seropositive. By contrast only 2:100 of the non-sexual contacts had evidence of infection. These were the daughter of an index case and her 2-year-old son who were seropositive but had their own independent risk factors. The data presented here support the hypothesis that a major route of transmission of HIV in an African population is through heterosexual contact and furthermore suggest that transmission by other means such as intimate non-sexual contact, insect vectors, or the use of non-sterile needles is much less important.

Acquired Immunodeficiency Syndrome↗