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Biomedical subjects

Ewert Linder

Publications and source records attributed to Ewert Linder.

8 recordsLinked to original sources

Overdiagnosis of Entamoeba histolytica and Entamoeba dispar in Nicaragua: a microscopic, triage parasite panel and PCR study.

BACKGROUND: Based on stool microscopy, an E. histolytica/E. dispar prevalence of 18.6% was found in León, Nicaragua about 10 years ago. Since then, new non-microscopic methods have been developed to discriminate between pathogenic E. histolytica and nonpathogenic E. dispar. The main objectives of the present study were to evaluate the true prevalence of E. histolytica among individuals with diarrhea and to assess the diagnostic procedures carried out at the health center level. METHODS: A descriptive study was carried out on patients with diarrhea. Parasite detection was performed by conventional microscopy on native preparations or concentrated and stained specimens, Triage Parasite Panel and by PCR for both E. histolytica and E. dispar. RESULTS: In 134 individuals with diarrhea, the prevalence of intestinal parasites was 69% as detected by direct stool examination. E. histolytica/E. dispar was found in eight (6%) of the samples, but the health centers reported 24%. In the Triage Parasite Panel only one case of E. histolytica/E. dispar was found. Analysis by PCR showed E. dispar in ten (7.5%) and E. histolytica in two cases (1.5%). The detection of intestinal coccidia and Dientamoeba fragilis required additional staining methods. CONCLUSIONS: PCR results showed that E. histolytica is a rare finding in patients with diarrhea. At the health centers, E. histolytica, E. histolytica/E. dispar were clearly overdiagnosed, with the consequence of overtreatment.

Animals↗

Schistosomiasis in Swedish travellers to sub-Saharan Africa: Can we rely on serology?

In the absence of egg excretion, laboratory diagnosis of recently acquired schistosomiasis is dependent on serology. 42 of 83 Swedish adventure tourists to sub-Saharan Africa had serum anti-schistosome antibodies indicating recent infection. There is little doubt regarding the specificity and sensitivity of serology for the demonstration of infection, but there is a need for alternative serological methods which could be more widely used than the standard immunofluorescence assay (IFA) for antibodies against gut-derived antigens (anti-GAA). We present results suggesting that 40/42 anti-GAA positive cases also react with keyhole limpet haemocyanin (KLH), a readily available commercial antigen. High anti-GAA titres were seen for more than 2 y despite treatment with praziquantel. Thus we are faced with several questions. How likely is it that positive serology means treatment failure? What is the risk involved in chronic infection? What is the prospect for monitoring treatment outcome by serology? We conclude that there is a need for better information on the risk of becoming infected, for improved methods for testing and for monitoring the therapeutic effects in adventure tourists.

Adolescent↗

Developmental changes in the adhesive disk during Giardia differentiation.

Giardia lamblia is a protozoan parasite infecting the upper mammalian small intestine. Infection relies upon the ability of the parasite to attach to the intestine via a unique cytoskeletal organelle, the ventral disk. We determined the composition and structure of the disk throughout the life cycle of the parasite and identified a new disk protein, SALP-1. SALP-1 is an immunodominant protein related to striated fiber-assemblin (SFA). The disk is disassembled during encystation and stored as four fragments in the immobile cyst. Serial Analysis of Gene Expression (SAGE) showed that the mRNA levels of the disk proteins decreased in encystation but two-dimensional protein gels showed that the protein levels were more constant. The parasite emerges without a functional disk but the four disk fragments are quickly reassembled into two new disks on the dividing, early excysting form. Thus, disk proteins are stored within the cyst, ready to be used in the rapid steps of excystation.

Amino Acid Sequence↗

Uptake and replication of Salmonella enterica in Acanthamoeba rhysodes.

The ability of salmonellae to become internalized and to survive and replicate in amoebae was evaluated by using three separate serovars of Salmonella enterica and five different isolates of axenic Acanthamoeba spp. In gentamicin protection assays, Salmonella enterica serovar Dublin was internalized more efficiently than Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium in all of the amoeba isolates tested. The bacteria appeared to be most efficiently internalized by Acanthamoeba rhysodes. Variations in bacterial growth conditions affected internalization efficiency, but this effect was not altered by inactivation of hilA, a key regulator in the expression of the invasion-associated Salmonella pathogenicity island 1. Microscopy of infected A. rhysodes revealed that S. enterica resided within vacuoles. Prolonged incubation resulted in a loss of intracellular bacteria associated with morphological changes and loss of amoebae. In part, these alterations were associated with hilA and the Salmonella virulence plasmid. The data show that Acanthamoeba spp. can differentiate between different serovars of salmonellae and that internalization is associated with cytotoxic effects mediated by defined Salmonella virulence loci.

Acanthamoeba↗

Antibodies in mother's milk protect children against giardiasis.

The protective effect of anti-Giardia antibodies in mother's milk on the acquisition of Giardia infection in their children during the first 2 y of life was analysed as part of a prospective study on infant diarrhoea in a group of 307 mothers and children in Leòn, Nicaragua. Among 24 children acquiring infection within the first 6 months, 23 were born to mothers lacking antibodies. These children also developed more severe diarrhoea. A significant difference between children born to mothers with and without antibodies with respect to the age at which the first Giardia infection was acquired was demonstrated by survival analysis and log rank test (p = 0.036). In conclusion, children born to non-immune mothers are at significantly higher risk of acquiring Giardia infection and developing giardiasis with more severe symptoms compared with children of immune mothers.

Animals↗

Localization and identification of Schistosoma mansoni/KLH-crossreactive components in infected mice.

KLH (Keyhole limpet hemocyanin) is highly immunogenic, and crossreactive epitopes occur widely in nature. In schistosomiasis, infected hosts generate antibodies reactive with KLH. This is of diagnostic importance but we lack detailed information on the immunogen-carrying molecules and their distribution in the worm. We used anti-KLH antibodies to localize cross-reacting epitopes in the various developmental stages of the parasite in experimental Schistosoma mansoni infection. The staining results show KLH crossreactivity in the life stages of the parasite. By immunoblotting we show that KLH-crossreactive antigenic epitopes in the parasite eggs are carbohydrates, also recognized by antibodies against soluble schistosome egg antigens. The localizations in the larval stages and in adult worms suggest that crossreacting antigenic epitopes are secretory products.

Animals↗

Free-living amoebae promote growth and survival of Helicobacter pylori.

Transmission routes of Helicobacter pylori remain poorly understood. The finding of bacterial DNA in water suggests the involvement of environmental factors. Here we report successful co-cultivation of H. pylori with Acanthamoeba castellanii, which circumvents the requirement of this bacterium for precise microaerobic conditions and a large supply of nutrients in order to grow. H. pylori was able to propagate and remain viable for several weeks in the presence of amoebae under experimental conditions. Intact, metabolically active bacteria could be demonstrated in vacuoles. The putative dependence of H. pylori on free-living amoebae in nature could be important with respect to transmission and prevalence, as shown for some other pathogenic bacteria.

Acanthamoeba↗

Use of recombinant cellulose-binding domains of Trichoderma reesei cellulase as a selective immunocytochemical marker for cellulose in protozoa.

Some unicellular organisms are able to encyst as a protective response to a harmful environment. The cyst wall usually contains chitin as its main structural constituent, but in some cases, as in Acanthamoeba, it consists of cellulose instead. Specific cytochemical differentiation between cellulose and chitin by microscopy has not been possible, due to the similarity of their constituent beta-1,4-linked hexose backbones. Thus, various fluorescent brightening agents and lectins bind to both cellulose and chitin. We have used a recombinant cellulose-binding protein consisting of two cellulose-binding domains (CBDs) from Trichoderma reesei cellulases linked together in combination with monoclonal anticellulase antibodies and anti-mouse immunoglobulin fluorescein conjugate to specifically stain cellulose in the cysts of Acanthamoeba strains for fluorescence microscopy imaging. Staining was observed in ruptured cysts and frozen sections of cysts but not in intact mature cysts. No staining reaction was observed with the chitin-containing cyst walls of Giardia intestinalis, Entamoeba dispar, or Pneumocystis carinii. Thus, the recombinant CBD can be used as a marker to distinguish between cellulose and chitin. Thirteen of 25 environmental or clinical isolates of amoebae reacted in the CBD binding assay. All 13 isolates were identified as Acanthamoeba spp. Five isolates of Hartmannella and seven isolates of Naegleria tested negative in the CBD binding assay. Whether cyst wall cellulose really is a unique property of Acanthamoeba spp. among free-living amoebae, as suggested by our findings, remains to be shown in more extensive studies.

Acanthamoeba↗