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J Flegr

Publications and source records attributed to J Flegr.

18 recordsLinked to original sources

Comparison of european Trichobilharzia species based on ITS1 and ITS2 sequences.

Schistosomes are parasites of considerable medical and veterinary importance and, therefore, all aspects of their biology have been intensively studied. In contrast, our knowledge of species of the largest genus, Trichobilharzia, is insufficient. Because morphological characterization of Trichobilharzia species provides a limited number of criteria for species determination, molecular data are required. In the present paper, we sequenced internal transcribed spacers ITS1 and ITS2, and 5.8S ribosomal RNA (rRNA) genes of 3 European Trichobilharzia species (T. regenti, T. szidati and T. franki). We showed that ITS1 and ITS2 sequences can be used in species identification. Repetitive elements were found in ITS1 of all 3 Trichobilharzia species; their number and length varied depending on the species. Phylogenetic analysis showed that the visceral T. franki is more related to the nasal T. regenti, than to the visceral T. szidati. The newly designed primer, which is specific for T. regenti, might be used as a tool for diagnosis of this potential pathogen.

Animals↗

Concordance between genetic relatedness and phenotypic similarities of Trichomonas vaginalis strains.

BACKGROUND: Despite the medical importance of trichomoniasis, little is known about the genetic relatedness of Trichomonas vaginalis strains with similar biological characteristics. Furthermore, the distribution of endobionts such as mycoplasmas or Trichomonas vaginalis virus (TVV) in the T. vaginalis metapopulation is poorly characterised. RESULTS: We assayed the relationship between 20 strains of T. vaginalis from 8 countries using the Random Amplified Polymorphic DNA (RAPD) analysis with 27 random primers. The genealogical tree was constructed and its bootstrap values were computed using the program FreeTree. Using the permutation tail probability tests we found that the topology of the tree reflected both the pattern of resistance to metronidazole (the major anti-trichomonal drug) (p < 0.01) and the pattern of infection of strains by mycoplasmas (p < 0.05). However, the tree did not reflect pattern of virulence, geographic origin or infection by TVV. Despite low bootstrap support for many branches, the significant clustering of strains with similar drug susceptibility suggests that the tree approaches the true genealogy of strains. The clustering of mycoplasma positive strains may be an experimental artifact, caused by shared RAPD characters which are dependent on the presence of mycoplasma DNA. CONCLUSIONS: Our results confirmed both the suitability of the RAPD technique for genealogical studies in T. vaginalis and previous conclusions on the relatedness of metronidazol resistant strains. However, our studies indicate that testing analysed strains for the presence of endobionts and assessment of the robustness of tree topologies by bootstrap analysis seem to be obligatory steps in such analyses.

Animals↗

Decrease of psychomotor performance in subjects with latent 'asymptomatic' toxoplasmosis.

Toxoplasma gondii is known to induce specific behavioural changes in its intermediate hosts. This is usually considered to be an evolutionary adaptation aimed to increase the probability of transmission of the parasite into its definitive host, the cat, by predation. In rodents an increase of reaction time as well as many other specific behavioural patterns have been observed. Here we report the results of our double blind study showing the significantly longer reaction times of 60 subjects with latent toxoplasmosis in comparison with those of 56 controls. Moreover, the existence of a positive correlation between length of infection and mean reaction time suggested that slow and cumulative effects of latent toxoplasmosis rather than a one-step (and possibly transient) effect of acute toxoplasmosis disease are responsible for the decrease of psychomotor performance of infected subjects. To our knowledge, this is the first study confirming the existence of such parasite-induced changes in human behaviour that could be considered in evolutionary history of the human species as adaptive from the point of view of parasite transmission.

Animals↗

Construction and bootstrap analysis of DNA fingerprinting-based phylogenetic trees with the freeware program FreeTree: application to trichomonad parasites.

The Win95/98/NT program FreeTree for computation of distance matrices and construction of phylogenetic or phenetic trees on the basis of random amplified polymorphic DNA (RAPD), RFLP and allozyme data is presented. In contrast to other similar software, the program FreeTree (available at http://www.natur.cuni.cz/~flegr/programs/freetree or http://ijs.sgmjournals.org/content/vol51/issue3/) can also assess the robustness of the tree topology by bootstrap, jackknife or operational taxonomic unit-jackknife analysis. Moreover, the program can be also used for the analysis of data obtained in several independent experiments performed with non-identical subsets of taxa. The function of the program was demonstrated by an analysis of RAPD data from 42 strains of 10 species of trichomonads. On the phylogenetic tree constructed using FreeTree, the high bootstrap values and short terminal branches for the Tritrichomonas foetus/suis 14-strain branch suggested relatively recent and probably clonal radiation of this species. At the same time, the relatively lower bootstrap values and long terminal branches for the Trichomonas vaginalis 20-strain branch suggested more ancient radiation of this species and the possible existence of genetic recombination (sexual reproduction) in this human pathogen. The low bootstrap values and the star-like topology of the whole Trichomonadidae tree confirm that the RAPD method is not suitable for phylogenetic analysis of protozoa at the level of higher taxa. It is proposed that the repeated bootstrap analysis should be an obligatory part of any RAPD study. It makes it possible to assess the reliability of the tree obtained and to adjust the amount of collected data (the number of random primers) to the amount of phylogenetic signals in the RAPD data of the taxon analysed. The FreeTree program makes such analysis possible.

Animals↗

Correlation of duration of latent Toxoplasma gondii infection with personality changes in women.

Many parasites induce characteristic changes in behavior of their hosts. In humans latent toxoplasmosis is associated with changes in personality profiles. It has been already shown that a decrease in superego strength is correlated with duration of toxoplasmosis in men. Here we studied changes in personality profiles with Cattell's 16 PF questionnaire in Toxoplasma-infected women. The changes were measured as differences in personality factors between Toxoplasma-infected subjects and uninfected controls of the same age. The low-rate changes were studied in 230 women diagnosed with acute toxoplasmosis during past 14 years. The results showed the correlation between duration of toxoplasmosis and level of factors G (high superego strength) and Q3 (high strength of self sentiment). The high-rate changes were estimated by measuring the correlation between level of Toxoplasma-antibody titers (which rapidly decline after the end of acute phase of toxoplasmosis) and personality factors in an experimental set of 55 young mothers with latent toxoplasmosis. Again, certain factors, namely A (affectothymia), F (surgence), G (high superego strength), H (parmia), and L (protension), correlated with the length of the infection. We suggest that the parasite induced the changes in the personality profiles of the women because of our observation of an increasingly different personality profile over time between women with latent infection and controls. The same evidence questions the view that women with a particular personality profile are more prone to acquisition of T. gondii infection.

Acute Disease↗

Transient nature of Toxoplasma gondii-induced behavioral changes in mice.

Many parasites induce specific changes in host behavior that promote the transmission of their infective stages between hosts. Toxoplasmosis in rodents is known to be accompanied by specific behavioral changes (shift in activity level, learning capacity, and novelty discrimination) that can theoretically increase the chance of infected animals being eaten by the definitive host, the cat. However, toxoplasmosis is also accompanied by many pathological symptoms. It is not known whether the behavioral changes are products of manipulation activity of the parasite or only nonspecific by-products of pathological symptoms of toxoplasmosis. Here, we compared the dynamics of development of behavioral and pathological changes in Toxoplasma gondii-infected mice. The results showed that the maximum reduction of mouse activity corresponded with the peak of pathological symptoms, and also that maximum increase of reaction times corresponded with the peak of development of tissue cysts in the brains of infected mice. Behavioral changes were only transient and disappeared before the 12th wk postinoculation. The results suggest that the behavioral changes in infected mice reported by many authors and observed in our experiments could be nonspecific by-products of pathological symptoms of toxoplasmosis rather than specific products of manipulation activity by the parasite.

Acute Disease↗

Changes in the personality profile of young women with latent toxoplasmosis.

Latent toxoplasmosis is the most widespread parasite infection in developed and developing countries. The prevalence of Toxoplasma gondii infection varies mostly between 20 to 80% in different territories. This form of toxoplasmosis is generally considered to be asymptomatic. Recently published results, however, suggest that the personality profiles of infected subjects differ from those of uninfected controls. These results, however, were obtained on non-standard populations (biologists or former acute toxoplasmosis patients). Here we studied the personality profiles of 191 young women tested for anti-Toxoplasma immunity during gravidity. The results showed that the differences between Toxoplasma-negative and Toxoplasma-positive subjects exits also in this sample of healthy women. The subjects with latent toxoplasmosis had higher intelligence, lower guilt proneness, and possibly also higher ergic tension. The difference in several other factors (desurgency/surgency, alaxia/protension, naiveté/shrewdness, and self-sentiment integration) concerned changes in the variances, rather than the mean values of the factors.

Adolescent↗

Free-Tree--freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap/jackknife analysis of the tree robustness. Application in the RAPD analysis of genus Frenkelia.

The Win95 program for computation of distance matrixes and construction of phylogenetic or phenetic trees on the basis of RAPD, RFLP and allozyme data was presented. In contrast with other presently available software, the program FreeTree can also assess the robustness of the tree topology by bootstrap, jackknife or OTU-jackknife analysis. Moreover, the program can be used also for an analysis of data obtained in several independent experiments performed with nonidentical subsets of taxa. The function of the program was demonstrated by an analysis of RAPD data from 22 strains of Frenkelia. The program is available as an autoextractive archive containing the installation files of FreeTree and TreeView, manual in MS Word format and a sample of the input file at http://www.natur.cuni.cz/flegr/programs/+ ++freetree.

Animals↗

Correlation between aerobic and anaerobic resistance to metronidazole in trichomonads: application of a new computer program for permutation tests.

An indispensable step of any comparative study is testing the concordance between the distribution of phenetic traits and the evolutionary history of the taxa under study. Here we present a computer program TREEPT which can perform these tasks on the basis of a permutation test. The use of the program was demonstrated on trichomonad drug sensitivity data. The program can also perform permutation tests analogous to the t-test, analysis of variance (ANOVA) and correlation analysis and is available at http://www.natur.cuni.cz/flegr/PROGRAMS/+ ++TREEPT.ZIP and http://www.karlin. mff.cuni.cz/-zaboj/treept.

Aerobiosis↗

The role of psychological factors in questionnaire-based studies on routes of human toxoplasmosis transmission.

The paper studies impacts of particular toxoplasmosis risk factors (consumption of raw meat and contact with cats), their interactions, and their relationship with the personality of the subjects. Among 243 men and 343 women the frequency of subjects with antitoxoplasma immunity was 26.6% and 21.6%, respectively. The association of antitoxoplasma immunity (assessed by the toxoplasmin skin test) with the two risk factors was estimated by log-linear analysis. Reported contact with cats has no influence on the probability of having antitoxoplasma immunity (P = 0.23) while the consumption of raw meat increased this probability (P = 0.0008). Very strong positive association between the contact with cats and the raw meat consumption was found among subjects without toxoplasmosis (P = 0.0028), suggesting that among these persons some subjects either incorrectly assessed their exposition to the risk factors or provided false data during the interview. The results of logistic regression suggest that the contact with cat and the consumption of raw meat are associated with particular personality traits. However, these traits differ from those associated with antitoxoplasma immunity suggesting that the correlation between antitoxoplasma immunity and consumption of raw meat reflects epidemiological importance of the raw meat rather than a correlation of both factors (raw meat consumption and probability of acquiring toxoplasmosis) with the subjects personality.

Adolescent↗

Induction of changes in human behaviour by the parasitic protozoan Toxoplasma gondii.

Toxoplasma gondii, the coccidian parasite, is known to induce changes in the behaviour of its intermediate hosts. The high prevalence of this parasite in the human population (20-80%) offers the opportunity of studying the influence of the parasite on human behaviour by screening of a normal population. A total of 224 men and 170 women were tested for toxoplasmosis and their personality profiles were measured by Cattell's questionnaire. Highly significant differences between Toxoplasma-infected and uninfected subjects were observed (P < 0.01). For men the factors G (low superego strength), L (protension), O (guilt proneness), and Q2 (group dependency) were positively influenced in infected subjects. For women the prevailing factors were A (affectothymia, P < 0.01), L (alaxia), O (untroubled adequacy) and Q2 (self-sufficiency). To reveal whether toxoplasmosis induces personality factor-shifts or whether certain combinations of personality factors influence the probability of acquiring Toxoplasma infection, we examined the personality profiles of 164 male patients diagnosed with acute toxoplasmosis during the past 13 years. The existence of a positive correlation between the duration of latent toxoplasmosis and the intensity of superego strength decrease (P < 0.02) suggested that the decrease of superego strength (the willingness to accept group moral standards) was induced by T. gondii infection.

Animals↗

Influence of chronic toxoplasmosis on some human personality factors.

An effect of parasites on host behaviour was tested on the toxoplasma-human model. Three hundred and thirty-eight (338) people were assessed with Cattell's personality questionnaire and then tested for Toxoplasma gondii infection with a delayed type hypersensitivity test for Toxoplasma. A highly significant correlation between chronic toxoplasmosis and two personality factors (G-Low Superego Strength and L-Protension) was found (p = 0.0032 and 0.0020, respectively). A correlation of the intensity of the personality factor-shifts with the duration of the infection (estimated from antibody titer) suggests that toxoplasmosis induces the shift in human personality, rather than the personality factors G and L influence an acquisition rate of Toxoplasma gondii infection.

Adolescent↗

Does a cell perform isoelectric focusing?

A model of intracellular electrical sorting of enzymes and organelles in the cytosol, based on isoelectric focusing, is proposed. The focusing is suggested to take place over a centrally symmetric pH gradient which in the cytosol of the yeast Saccharomyces cerevisiae is known to be 7.2-6.4. From published data on the energetic capacity and from the computed electric resistance of the S. cerevisiae cell, the maximum value of the electric field that can be maintained in the cytosol was estimated. The results showed that the strength of a centrally symmetric intracytosolic electric field could be as high as 90 mV/cm, which is sufficient to account for sorting of cytosolic proteins according to their isoelectric points. Although direct experimental evidence for intracellular isoelectric focusing is still missing, several phenomena of physiological importance can be understood on the assumption of its real existence.

Animals↗

Multiple populations of double-stranded RNA in two virus-harbouring strains of Trichomonas vaginalis.

The existence of six dsRNA segments of Trichomonas vaginalis virus was confirmed and the molar mass and relative abundance of these segments were determined by agarose gel electrophoresis with reovirus dsRNA serving as a standard. The M's were 3.5, 3.4, 3.2, 2.5, 1.4 and 0.34 Mg/mol for the two strains studied, the relative abundances, however, were 1.0, 1.4, 3.0, 0.3, 2.7, 4.2 and 1.0, 0.6, 1.7, 0.5, 3.4 1.0 for these strains, respectively. Cell homogenate fractionation showed that all dsRNA segments were associated with viral particles. The data appeared to support the hypothesis of a relationship between viruses of the protozoan T. vaginalis and of the yeast Saccharomyces cerevisiae.

Animals↗

The dsRNA of Trichomonas vaginalis is associated with virus-like particles and does not correlate with metronidazole resistance.

Twelve metronidazole-resistant and twelve metronidazole-susceptible strains of Trichomonas vaginalis were tested for the presence of dsRNA. Three resistant and five susceptible strains were found to contain dsRNA which indicated that metronidazole resistance does not correlate with the absence of dsRNA. Electron microscopy showed the homogenates of all dsRNA-positive strains to contain virus-like particles 32-38 nm in diameter, while no such particles were found in the dsRNA-negative strains. A mutual relationship between the dsRNA and virus-like particles seems to exist.

Animals↗

A rapid method for isolation of double stranded RNA.

A rapid and simple method for the isolation and purification of dsRNA is presented. The crucial step of this method is the extraction of proteins and DNA with acid phenol. After the extraction, only RNA is left in the aqueous phase. ssRNA contamination of the RNA preparation can be greatly reduced when ammonium sulfate is present during the extraction.

Animals↗

Characterization of trichomonad species and strains by PCR fingerprinting.

The random amplified polymorphic DNA (RAPD) technique was used for phylogenetic analysis of trichomonads, for intraspecies genealogical study of Trichomonas vaginalis strains, and for assessment of intrastrain polymorphism in Trichomonas vaginalis. The phylogenetic tree for 12 trichomonad species showed certain discrepancies with current models of trichomonad evolution. However, it shows that RAPD traits retain phylogenetically relevant information. The results of intraspecies analyses of 18 Trichomonas vaginalis strains suggested some concordance between the genetic relationship of strains and their geographic origin. They also suggested a concordance between the strain genetic relationships and the resistance to metronidazole. A concordance was also found with respect to the severity of disease observed in donor patients but not with the results of laboratory virulence assays. No concordance was found between genetic relationship of strains and strain infection with a dsRNA Trichomonas vaginalis virus (TVV). The latter suggests that TVV might be transmitted horizontally among Trichomonas vaginalis populations. The identity of RAPD patterns of clones isolated from in vitro cultures and those of the cultures reisolated independently from the same patient within a period of six weeks suggests that individual Trichomonas vaginalis strains are not polymorphic and that the RAPD patterns are stable. Therefore, the RAPD technique seems useful for addressing various clinically relevant issues.

Animals↗

Molecular phylogenetic relatedness of Frenkelia spp. (Protozoa, Apicomplexa) to Sarcocystis falcatula Stiles 1893: is the genus Sarcocystis paraphyletic?

The coccidians Frenkelia microti and F. glareoli (Apicomplexa: Sarcocystidae) form tissue cysts in the brain of small rodents (intermediate hosts) while oocysts are formed in the intestine of final hosts, buzzards of the genus Buteo. The inclusion of the small subunit ribosomal RNA gene sequences (SSU rRNA) of both Frenkelia species into the SSU rRNA trees of other, tissue cyst-forming coccidia strongly supports paraphyly of the genus Sarcocystis. Frenkelia spp. exhibit close relatedness to Sarcocystis falcatula Stiles 1893, a bird-opossum parasite, recognized under its junior synonym S. neurona Dubey et al. 1991, as the causative agent of equine protozoan myeloencephalitis on the American continent. As the definition of the genus Frenkelia is based on a plesiomorphic character (affinity to the neural tissue) of supposedly low phylogenetic value, the synonymization of the genus Frenkelia with Sarcocystis is proposed. This renders the genus Sarcocystis monophyletic.

Animals↗