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PubMed1991

The effect of ionizing radiation on the viability of Trichomonas vaginalis.

1. The effects of continuous gamma radiation on the viability of Trichomonas vaginalis (ATCC 30001) were assessed by a colony count technique. 2. A triphasic survival curve showed an initial shoulder (Dq) of 3 Gy followed by three linear curves with D0 values of 34, 300, and 90 Gy. 3. Sterilization of 10(6) cells/ml occurred from 1600 to 1800 Gy of radiation. 4. Population growth, subsequent to radiation exposure of 17-100 Gy, showed an increased lag time followed by a faster rate of growth, compared with unirradiated cells. 5. Trichomonas vaginalis is more sensitive to ionizing radiation than free-living protozoa and appears as radiosensitive as those parasitic protozoa examined in radioattenuation experiments.

Animals↗
PubMed1995

Sequence of cDNA coding for a 65 kDa adhesive protein for the specific detection of Trichomonas vaginalis by PCR.

A Trichomonas vaginalis cDNA library was constructed and recombinant plaques were screened using rabbit immunoglobulins specific for P65, a protozoan protein involved in pathogenicity that we identified in a previous study. A 1.38 kilobases cDNA fragment coding for the P65 protein was cloned in E. coli and then sequenced. On the basis of of the sequence obtained, six primers were synthesised and used to set up a Polymerase Chain Reaction. The presence of a specific amplicon in all 30 clinical isolates tested shows that P65 is a conserved and stable gene. The reaction is highly sensitive (as few as 5 to 10 parasites can be detected) and specific for Trichomonas vaginalis; the gene coding P65 adhesin can be therefore considered a very good molecular target for polymerase chain reaction-based diagnostic purposes.

Animals↗
PubMed2005

Rubrerythrin and peroxiredoxin: two novel putative peroxidases in the hydrogenosomes of the microaerophilic protozoon Trichomonas vaginalis.

The parasitic flagellate Trichomonas vaginalis contains hydrogenosomes, anaerobic organelles related to mitochondria, that generate ATP from the fermentative conversion of pyruvate to acetate, CO2 and molecular hydrogen. Although an essentially anaerobic organism, Trichomonas encounters low oxygen concentrations in its natural habitat and has to protect itself, and especially the oxygen-sensitve enzymes of hydrogenosomal metabolism, from oxidative damage. We have identified two novel proteins in the hydrogenosomal proteome with strong similarity to two putative prokaryotic peroxidases, rubrerythrin and periplasmic thiol peroxidase. Both proteins have previously been found in many prokaryotes but were not known from eukaryotes, suggesting a significant prokaryotic component in the oxygen-detoxification system of trichomonad hydrogenosomes.

Amino Acid Sequence↗
PubMed2005

Identification of a very large Rab GTPase family in the parasitic protozoan Trichomonas vaginalis.

Rab proteins are pivotal components of the membrane trafficking machinery in all eukaryotes. Distinct Rab proteins locate to specific endomembrane compartments and genomic studies suggest that Rab gene diversity correlates with endomembrane system complexity; for example unicellular organisms generally possess 5-20 Rab family members and the size of the repertoire increases to 25-60 in multicellular systems. Here we report 65 open reading frames from the unicellular protozoan Trichomonas vaginalis that encode distinct Rab proteins (TvRabs), indicating a family with complexity that rivals Homo sapiens in number. The detection of gene transcripts for the majority of these genes and conservation of functional motifs strongly suggests that TvRabs retain functionality and likely roles in membrane trafficking. The T. vaginalis Rab family includes orthologues of the conserved subfamilies, Rab1, Rab5, Rab6, Rab7 and Rab11, but the majority of TvRabs are not represented by orthologues in other systems and includes six novel T. vaginalis specific Rab subfamilies (A-F). The extreme size of the T. vaginalis Rab family, the presence of novel subfamilies plus the divergent nature of many TvRab sequences suggest both the presence of a highly complex endomembrane system within Trichomonas and potentially novel Rab functionality. A family of more than 65 Rab genes in a unicellular genome is unexpected, but may be a requirement for progression though an amoeboid life-cycle phase as both Dictyostelium discoideum and Entamoeba histolytica share with T. vaginalis both an amoeboid life cycle stage and very large Rab gene families.

Amino Acid Sequence↗
PubMed2005

A high-throughput colorimetric and fluorometric microassay for the evaluation of nitroimidazole derivatives anti-trichomonas activity.

Several nitroimidazole derivatives were synthesized and tested as possible trichomonicidal agents. A fast, simple, practical and reliable in vitro colorimetric method was applied to the screening of the nitroimidazole derivatives anti-trichomonas activity. The colorimetric technique was based on the use of Alamar blue as a redox-indicator. The test was carried out both qualitatively (minimal inhibitory concentration determined by naked eye observation) and quantitatively (fluorometric determination of 50% and 90% inhibitory concentrations), the latter took advantage of the dye fluorometric properties. The performance of the method was excellent affording an exactitude 97.86% and a reproducibility of 95% and no interference of the trichomonas-culture medium was observed during the test. Some of the nitroimidazole compounds tested showed a fair trichomonicidal activity, however none of them was as active as the model compound, metronidazole.

Animals↗
PubMed2003

Molecular biology of the amitochondriate parasites, Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis.

The amitochondriates are an assembly of unicellular protists that lack mitochondria, and often other typical eukaryotic organelles, such as peroxisomes. Relatively little research has been conducted on amitochondriates, even though marine waters are likely to be teeming with such organisms, representing yet unknown protist lineages. The three amitochondriates that have been studied in any detail are the three mucosal human parasites: Giardia, Trichomonas and Entamoeba. These pathogens have worldwide distribution and are the most commonly encountered parasites in North America and Europe. Despite significant differences in their lifecycles and pathogenic properties, Giardia, Trichomonas and Entamoeba are customarily grouped together based on their being microaerophilic, their anaerobic carbohydrate metabolism, their lack of mitochondria, and their placement on deep-branching lineages in eukaryotic phylogenetic trees. During the last decade, the development of functional tools has allowed molecular analyses of gene expression to be initiated on these divergent eukaryotes. The resulting data indicate significant differences between the organisation of genetic information and mechanisms of gene regulation in amitochondriates and other organisms. Promoter architecture, as well as the regulatory transcription factors required to mediate promoter activity, lacks the conservation observed for the transcriptional apparatuses of metazoa and even appear to be unique within the amitochondriates. Our knowledge of the molecular biology and gene expression in amitochondriates is still in its infancy and a discussion of the current status of research in this area is presented.

3' Untranslated Regions↗
PubMed2003

Biochemical and genetic evidence for a family of heterotrimeric G-proteins in Trichomonas vaginalis.

We have cloned a single copy gene from the human parasite Trichomonas vaginalis that encodes a putative protein of 402 amino acids with approximately 35% sequence identity to known alpha subunits of heterotrimeric G-proteins. It contains the characteristic GTP binding domains G-1 to G-5 with the key residues conserved. The new sequence has an unusual N-terminal extension of approximately 70 residues that cannot be aligned to reference G-proteins and which is characterised by proline-rich repeats. To investigate the expression and cellular localisation of the protein we produced specific antisera against a recombinant fusion protein. The antisera recognised a protein of an apparent molecular mass of 51 kDa in protein extracts from T. vaginalis and immunofluorescent microscopy established that the protein is localised to discrete endomembranes. Using a protocol designed to purify mammalian heterotrimeric G-proteins incorporating a GTPgammaS binding assay, we isolated two proteins from Trichomonas that are recognised by an heterologous GA/1 antisera raised to a peptide of the conserved G-1 domain of G-protein alpha subunits. These two proteins have an apparent molecular mass of 61 and 48 kDa, respectively, larger and smaller than the translation product of the cloned gene. Consistent with these results, the GA/1 antisera did not cross-react with the fusion protein produced from the gene we have cloned. These data suggest T. vaginalis possesses more than one heterotrimeric G-protein alpha subunit. Based on the sequence features of the cloned gene and the biochemical properties of the purified proteins, we suggest that these alpha subunits are likely to be part of classic heterotrimeric G-protein complexes.

Amino Acid Sequence↗
PubMed2000

In vitro screening of american plant extracts on Trypanosoma cruzi and trichomonas vaginalis.

From the beginning of this decade and with the revival of the phytotherapy, biological research about immunomodulatory, anti-inflammatory and antiprotozoal effects of Central and South American plants have been in progress. Our objective was to determine the antiprotozoal activity of 79 extracts from different plant families, including Asteraceae, Araceae, Moraceae, Solanaceae, Rhamnaceae, Zingiberaceae, Leguminosae and Sapotaceae. Once matching with herbarium specimens authenticated the plants, selected parts were separated, dried carefully and reduced to powder. Most of the screened extracts were aqueous. Two protozoa with different metabolic pathways, Trypanosoma cruzi and Trichomonas vaginalis were used as experimental models. Trypanocidal activity of plants was assayed on epimastigote cultures in liver infusion tryptose (LIT). Anti-Trichomonas activity was determined over cultures of the parasite in Diamond medium. In both cases, microscopic counting of parasites, after their incubation in the presence of different concentrations of the crude extracts, were made in order to determine the cytocidal and cytostatic activities respect to control cultures. Of the nine extracts that showed antiprotozoal activity, those from Mikania cordifolia and Philodendron bipinnatifidum were then fractionated, and again, were assayed the organic and aqueous phases obtained.

Americas↗
PubMed1991

Peritonitis caused by Trichomonas--a preliminary case report.

Trichomonas infections of the abdominal cavity are uncommon and case histories have not been published previously. A patient to whom reference is made this paper developed a combined immunodeficiency syndrome and portal hypertension with ascites on the basis of a cirrhosis of the liver. This seems to have been the precondition for a peritonitis caused by a Trichomonas species.

Ascites↗
PubMed1991

In vitro effects of berberine sulphate on the growth and structure of Entamoeba histolytica, Giardia lamblia and Trichomonas vaginalis.

The plant alkaloid, berberine sulphate, inhibited the growth of Entamoeba histolytica, Giardia lamblia and Trichomonas vaginalis in BI-S-33 medium, and induced morphological changes in the parasites. Exposure of E. histolytica to berberine caused a clumping of chromatin in the nucleus, and the formation of autophagic vacuoles and aggregates of small vacuoles in the cytoplasm. In berberine-treated G. lamblia, an irregularly-shaped vacuole appeared in the cytoplasm and gradually enlarged during culture. The trophozoites became swollen and deposits of glycogen were seen in the cytoplasm. Trichomonas vaginalis was also affected by the berberine; autophagic vacuoles increased in number soon after exposure, and one large vacuole, which was characteristic of treated cells, appeared. These observations demonstrate that, in vitro, this drug was effective against E. histolytica, G. lamblia and T. vaginalis. Another alkaloid, coptisine, was inactive against the three parasites.

Animals↗
PubMed1994

Is Trichomonas vaginalis a cause of cervical neoplasia? Results from a combined analysis of 24 studies.

BACKGROUND: We conducted this combined analysis of available data from studies with information on this issue to clarify the association between Trichomonas vaginalis infection and cervical neoplasia. METHODS: We performed MEDLINE searches (1966-1993) using the key words and phrases 'trichomonas vaginitis' and 'neoplasms, cervix' for articles published in English, and searched citations of the articles obtained from MEDLINE: A total of 24 articles (two cohort studies and 22 case-control) were included in this data analysis. In the analysis, the studies were evaluated for heterogeneity using Breslow-Day tests for homogeneity of the odds ratios and of rate ratios. If the odds ratios from studies are heterogeneous, it is not appropriate to combine them using the Mantel-Haenszel method. Also, publication bias was evaluated by assessing the association between the observed effect size and the variance of the effect size using a rank correlation test. RESULTS: The combined summary relative risk for the two cohort studies was 1.93 (95% confidence interval: 1.22-2.65) indicating an approximate doubling of the risk of cervical neoplasia in the presence of T. vaginalis infection. The attributable risks among exposed subjects and among the source population were 47.4% and 2.1% respectively. Results of the 22 retrospective studies were much less consistent. However, most of them demonstrated a significant positive association. CONCLUSIONS: This combined analysis suggests that there is an association between T. vaginalis and the risk of cervical neoplasia, but that such infections account for only 2% of cervical neoplasia.

Adolescent↗
PubMed2001

Mitochondrial type iron-sulfur cluster assembly in the amitochondriate eukaryotes Trichomonas vaginalis and Giardia intestinalis, as indicated by the phylogeny of IscS.

Pyridoxal-5'-phosphate-dependent cysteine desulfurase (IscS) is an essential enzyme in the assembly of FeS clusters in bacteria as well as in the mitochondria of eukaryotes. Although FeS proteins are particularly important for the energy metabolism of amitochondrial anaerobic eukaryotes, there is no information about FeS cluster formation in these organisms. We identified and sequenced two IscS homologs of Trichomonas vaginalis (TviscS-1 and TviscS-2) and one of Giardia intestinalis (GiiscS). TviscS-1, TviscS-2, and GiiscS possess the typical conserved regions implicated in cysteine desulfurase activity. N-termini of TviscS-1 and TviscS-2 possess eight amino acid extensions, which resemble the N-terminal presequences that target proteins to hydrogenosomes in trichomonads. No presequence was evident in GiiscS from Giardia, an organism that apparently lacks hydrogenosmes or mitochondria. Phylogenetic analysis showed a close relationship among all eukaryotic IscS genes including those of amitochondriates. IscS of proteobacteria formed a sister group to the eukaryotic clade, suggesting that isc-related genes were present in the proteobacterial endosymbiotic ancestor of mitochondria and hydrogenosomes. NifS genes of nitrogen-fixing bacteria, which are IscS homologs required for specific formation of FeS clusters in nitrogenase, formed a more distant group. The phylogeny indicates the presence of a common mechanism for FeS cluster formation in mitochondriates as well as in amitochondriate eukaryotes. Furthermore, the analyses support a common origin of Trichomonas hydrogenosomes and mitochondria, as well as secondary loss of mitochondrion/hydrogenosome-like organelles in Giardia.

Amino Acid Sequence↗
PubMedDate not supplied

Inhibition of Trichomonas vaginalis by fungi during associative growth.

In controlled studies of associative growth, Candida albicans and Torulopsis glabrata, but not Candida parapsilosis, inhibited the proliferation and eventually the viability of Trichomonas vaginalis. The inhibition patterns produced against the trichomonad by C. albicans or T. glabrata were observed with trichomonad-fungus inoculum ratios ranging from 230:1 to 1:100. The onset of inhibition of T. vaginalis by T. glabrata was immediate, whereas that of C. albicans began after incubation for 48 hr. Under routine clinical conditions, T. vaginalis was identified by a combination of direct microscopy and culture in specimens from 330 (19.5%) of 1,691 women with vaginal discharges who were examined. Trichomonas vaginalis was isolated from 49 of 53 vaginal discharge specimens that also contained C. albicans. Therefore, the qualitative results with clinical cultures did not confirm the inhibition of trichomonads by C. albicans as shown in quantitative studies of associative growth. This result indicates that with the routine clinical isolation procedures the use of an antifungal agent in medium for culture of T. vaginalis is not necessary.

Candida↗
PubMed2002

Mechanisms of in vitro development of resistance to metronidazole in Trichomonas vaginalis.

Development of resistance against metronidazole and mechanisms responsible for this process were studied in a sexually transmitted pathogen of humans, Trichomonas vaginalis. Monitoring of changes in metabolism and protein expression that accompanied increasing resistance of strains derived from a common drug-susceptible parent (TV 10-02) showed the multistep character of the process. The aerobic type of resistance known to occur in isolates from patients non-responsive to treatment appeared at the earliest stage, followed by development of the anaerobic type of resistance which was accompanied by gradual loss of hydrogenosomal proteins associated with drug-activating pathways [pyruvate:ferredoxin oxidoreductase (PFOR), hydrogenase, ferredoxin]. Unexpectedly, the loss of PFOR did not result in acquisition of full anaerobic resistance, thus indicating an alternative source of electrons required for the drug activation. These data suggest involvement of the oxidative decarboxylation of malate in hydrogenosomes, catalysed by NAD(+)-dependent malic enzyme and subsequent transfer of reduced equivalents to the drug via NADH:ferredoxin oxidoreductase and ferredoxin. Accordingly, all components of this pathway were eliminated before the resistance was fully developed. Resistant Trichomonas vaginalis compensated the impaired function of hydrogenosomes by enhanced conversion of pyruvate to lactate in the cytosol. Further analysis of the two key enzymes involved in metronidazole activation by Northern blotting and assay for nascent mRNA showed that the insufficient expression of the PFOR protein results from decreased gene transcription, while down-regulation of malic enzyme is controlled at the mRNA level.

Animals↗
PubMed2004

Production of ammonia by Tritrichomonas foetus and Trichomonas vaginalis.

Production of ammonia is difficult to find among the various studies of amino acid metabolism in protozoa. Several studies suggest that catabolism of arginine to ammonium is important for the growth of trichomonads. Trichomonads are amitochondriate zooflagellates that thrive under microaerophilic and anaerobic conditions. The authors were able to detect accumulation of ammonium ions and ammonia in cultures of Tritrichomonas foetus and Trichomonas vaginalis, including those resistant to metronidazole. Ammonium ions and ammonia were detected using the indophenol colorimetric method. Cells incubated overnight under an ambient oxygen gas phase had 0.9 mM soluble ammonium (NH(4)(+) and NH(3)) or a 20 % greater concentration of ammonium relative to sterile growth medium that had been incubated similarly. Production of ammonia itself was confirmed by analysis of a wick that was moistened with sulfuric acid (20 mM) and placed above the liquid in sealed cultures of a strain of Trichomonas vaginalis. The wicks from these cultures captured the equivalent of 0.048 mM volatile ammonia (NH(3)) from the liquid as compared to 0.021 mM volatile ammonia from sterile medium after overnight incubation. Intact trichomonads, 0.7 x 10(6) cells ml(-1) equivalent to 0.7 mg protein ml(-1), incubated in Doran's buffer with or without (1 mM) L-arginine produced significant amounts of soluble ammonium (0.07 mM and 0.04 mM, respectively) during 60 min. The results indicate that ammonium ions and the more irritating ammonia are significant metabolites of trichomonads. In addition, based upon end-product amounts, it appears that the rate of arginine metabolism is of the same order of magnitude as that for carbohydrate metabolism by trichomonads.

Ammonia↗
PubMed1997

Use of inhibitors to identify essential cysteine proteinases of Trichomonas vaginalis.

Designing cysteine proteinase inhibitors as antitrichomonal drugs requires knowledge of which cysteine proteinases are essential to the parasite. In an attempt to obtain such information, the effects of a number of cysteine proteinase inhibitors on trichomonad growth in vitro and proteinase activity were investigated. The broad specificity inhibitor trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (known as E-64) had little effect on growth of Trichomonas vaginalis (27% inhibition at 280 microM, none at 28 microM) even though the addition of 2.8 microM E-64 to growth medium resulted in inhibition of all but two (apparent molecular masses: 35 k and 49 k) of the parasite's proteinases detected by gelatin SDS-PAGE. This shows that many of the parasite's cysteine proteinases are not essential for growth in axenic culture. In contrast, a peptidyl acyloxymethyl ketone, N-benzoyloxycarbonyl-Phe-Ala-CH2OCO-(2,6,-(CF3)2)Ph, at 16 microM killed T. vaginalis and severely inhibited growth of Tritrichomonas foetus. Exposure of Trichomonas vaginalis to 16 microM of this compound for 1 h resulted in both the 35 kDa and 49 kDa proteinases being inhibited, whereas some other proteinases were unaffected. Similar distinctions between the inhibitor sensitivity of the parasite's cysteine proteinases were apparent when a biotinylated peptidyl diazomethyl ketone was used to detect active proteinases. These data suggest that the growth inhibitory effects of the peptidyl acyloxymethyl ketone are through inhibition of cysteine proteinases that are not affected when the parasites are grown in the presence of E-64. At least one of these enzymes, which include the 35 kDa and 49 kDa cysteine proteinases, must be essential and so a suitable target for chemotherapeutic attack.

Animals↗
PubMed1984

Specific and common antigens of Trichomonas vaginalis detected by monoclonal antibodies.

Monoclonal antibodies to Trichomonas vaginalis were prepared by immunizing mice with a cloned isolate of T. vaginalis. Eight antibodies reacted with the same four isolates or strains but did not react with the other T. vaginalis strains or isolates tested. All eight antibodies reacted uniformly with both the body and flagella of T. vaginalis in the immunofluorescence assay but were unreactive by immunoblotting. The antigen(s) recognized by these antibodies was determined to be present on the surface membrane by indirect immunofluorescence assay of live organisms. The antigen(s) was found to be sensitive to periodate oxidation but resistant to pronase digestion. In addition, one monoclonal antibody was derived which reacted with all T. vaginalis isolates or strains tested, as well as with Trichomonas gallinae, Tritrichomonas foetus, and Giardia lamblia. This antibody reacted with the body but not the flagella of Formalin-fixed protozoa in the immunofluorescence assay but failed to react with live organisms. The antigen was found to be periodate resistant but pronase labile. In the immunoblot assay, this antibody detected a single T. vaginalis polypeptide with a molecular weight of 62,000.

Animals↗
PubMed1985

Specific nature of Trichomonas vaginalis parasitism of host cell surfaces.

The adherence of Trichomonas vaginalis NYH 286 to host cells was evaluated by using monolayer cultures of HeLa and HEp-2 epithelial cells and human fibroblast cell lines. Saturation of sites on HeLa cells was achieved, yielding a maximal T. vaginalis NYH 286-to-cell ratio of two. The ability of radiolabeled NYH 286 to compete with unlabeled trichomonads for attachment and the time, temperature, and pH-dependent nature of host cell parasitism reinforced the idea of specific parasite-cell associations. Other trichomonal isolates (JH31A, RU375, and JHHR) were also found to adhere to cell monolayers, albeit to different degrees, and all isolates produced maximal contact-dependent HeLa cell cytotoxicity. The avirulent trichomonad, Trichomonas tenax, did not adhere to cell monolayers and did not cause host cell damage. Interestingly, parasite cytadherence was greater with HeLa and HEp-2 epithelial cells than with fibroblast cells. In addition, cytotoxicity with fibroblast cells never exceeded 20% of the level of cell killing observed for epithelial cells. Elucidation of properties of the pathogenic human trichomonads that allowed for host cell surface parasitism was also attempted. Treatment of motile T. vaginalis NYH 286 with trypsin diminished cell parasitism. Incubation of trypsinized organisms in growth medium allowed for regeneration of trichomonal adherence, and cycloheximide inhibited the regeneration of attachment. Organisms poisoned with metronidazole or iodoacetate failed to attach to host cells, and adherent trichomonads exposed to metronidazole or iodoacetate were readily released from parasitized cells. Coincubation experiments with polycationic proteins and sugars and pretreatment of parasites or cells with neuraminidase or periodate had no effect on host cell parasitism. Colchicine and cytochalasin B, however, did produce some inhibition of adherence to HeLa cells. The data suggest that metabolizing T. vaginalis adheres to host cells via parasite surface proteins in a specific receptor-ligand fashion. Furthermore, parasitism of epithelial cells appears to render this cell type more susceptible than fibroblast cell types to contact-dependent cytotoxicity.

Adhesiveness↗