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[Occurrence of the protozoa, Entamoeba gingivalis and Trichomonas tenax in the mouths of children and adolescents with hyperplastic gingivitis caused by phenytoin].

The oral protozoa Entamoeba gingivalis and Trichomonas tenax do not occur in small children and are rarely found in older ones. In adolescents their occurrence rate keeps increasing with age. They parasitize in an oral cavity changed by inflammation, yet also in a healthy mouth. Their highest occurrence rate has been recorded in adults with periodontosis and atrophy of the periodontium, a somewhat lower one in adults with gingivitis. The authors addressed the question whether the presumed low occurrence rate of oral protozoa in children becomes increased in drug-induced gingivitis after treatment with the antiepileptic 5,5-diphenylhydantoin. Cultivation for oral protozoa was performed in 231 children and adolescents. Of these 59 were epileptics. Drug-induced gingivitis was present in 66% of the epileptics. Drug-induced gingivitis did not increase the occurrence rate of oral protozoa as compared to the findings in the rest of the series studied. (Tab. 9, Ref. 7.).

Adolescent↗

[Paraendocervical route for metronidazole administration in complex treatment of chronic Trichomonas endocervicitis].

The data on paraendocervical administration of metronidazole in the treatment of patients with relapsing Trichomonas endocervicitis are presented. Metronidazole was administered as 0.5% solution in a dose of 0.04 g once a day for 8-10 days in complex traditional therapy including oral use of metronidazole and immunocorrigating and local treatment. It was shown that paraendocervical administration of the protistocidal agent provided earlier regression of the urogenital symptoms and 2.8 times lower frequency of the relapses.

Administration, Intravaginal↗

Cryopreservation of Trichomonas vaginalis: a trial of using four different cryoprotectants.

To reach the maximum survival rate of Trichomonas vaginalis after cryopreservation, four substances: dimethylsulphoxide (DMSO), glycerol, egg yolk and horse serum were used as cryoprotectants. T. vaginalis trophozoites, in the logarithmic phase, cultured on TYM-S medium were used. Rapid and slow freezing schedules were followed. Examinations for viable organisms were done after a month duration in liquid nitrogen. Immediate and delayed thawing were tried. Viability scores depended on the presence of live organisms in the thawed parasite suspensions determined by the presence of motile trophozoites under microscopic examination, by dye uptake and by re-suspension of the organisms in fresh TYM-S and watching for parasite growth and multiplication. Rapid freezing protocol failed to cryopreserve T. vaginalis with any cryoprotectant. DMSO was superior to glycerol especially in 10% conc. by slow freezing. Viable organisms were more when the suspensions were immediately thawed without delay.

Animals↗

[Study on surface adhesion protein 33 gene sequence of different Trichomonas vaginalis isolates].

OBJECTIVE: To study genetic polymorphism of surface adhesion protein 33 (AP33) gene on the seven isolates of Trichomonas vaginalis. METHODS: PCR technique was performed to amplify AP33 gene from the seven isolates, DNA sequences were obtained from the AP33 gene of the isolates and phylogenetic tree was built. Minimal lethal concentrations (MLC) of metronidazole on the isolates were measured in vitro. RESULTS: Percentage of the similarity between 7 isolates and U87098 in GenBank was 98.2%-100%, which indicated a high homology and belonged to isotype isolates. There were four branches between Beijing 1 isolate and Tangshan isolate, Jiujiang 1 isolate and Jiujiang 2 isolate, Beijing 2 isolate and Jiujiang 3 isolate, Chengde isolate and U87098 isolate in phylogenetic tree, which showed a close genetic relationship respectively. No relativity was detected between geographical origin and genetic relationship. CONCLUSION: There is a close genetic relationship among the seven T. vaginalis isolates. MLC showed a difference between isolates which have close relationship.

Animals↗

Molecular cloning and characterization of a Rac1 homologue cDNA from Trichomonas vaginalis.

OBJECTIVE: To clone and characterize a Rac1 homologue from Trichomonas vaginalis for studying cell cycle of the organism. METHODS: A cDNA library derived from T. vaginalis mRNA was constructed into lambda TriplEx2 phage vector. An expression sequence tag program was launched. Sequences of cDNA clones were analyzed using NCBI BLAST algorithms, and ClustalW and Treeview programs. RESULTS: A cDNA clone with a length of 714 base pairs was isolated. The sequence analysis showed that the cDNA clone has an open reading frame with 600 bp. The deduced amino acid sequence from the open reading frame contains 200 residuals and is most homologous to Rac1 subfamily of Rho GTPases with >60% identity. The conserved sequence elements of Rho GTPases, such as GTP-binding sites, GTPase-activating protein (GAP) interaction motifs, GTP-dissociation inhibitors (GDI) interaction motifs, guanine nucleotide exchange factor (GEF) interaction elements, etc, were detected in the amino acid sequence. The phylogenetic analysis showed that the cDNA clone is grouped in the Rac subfamily and is more closely related to Rac1 proteins of protozoa. CONCLUSION: The cDNA clone isolated belongs to Rac subfamily of Rho GTPases and is probably a Rac1 protein of T. vaginalis.

Amino Acid Sequence↗

[In vivo observations of the division of the oversized round forms of Trichomonas vaginalis in culture].

Continuous in vivo observations of the behaviour of multinucleate, multiflagellate forms of Trichomonas vaginalis have been made in a microchamber at 36 degrees C. It has been proven that these forms are initial forms of the multiplication of the parasite by cell division. Serial in vivo shots have been taken in this microchamber of the budding of a young flagellate from a multinucleate, multiflagellate form. In vivo observations have shown that these round forms are multiplicable, living cell forms of the parasite.

Animals↗

[Studies on the susceptibility of Trichomonas hominis to some abiotic factors. I. Preliminary assessment of antitrichomonal activity of selected compounds in vitro].

The flagellate Trichomonas hominis is often identified in human diarrheic stools. Because of the fecal-oral transmission rout, infection with T. hominis is more frequently reported in children than in adults. Although infections with the trichomonads in some world regions are as often as with Giardia, no prevention and optimal treatment are defined. The purpose of our in vitro studies was to examine susceptibility of T. hominis to a newly synthesized imidazole derivative--1-metylo-2-(2,4-dinitrobenzylotio)-1H-imidazol--P27; in order to make a comparison between this new and previously known compounds, chlorhexidine and metronidazole were used. The protozoans derived from the diarrheic stool of adult patient were grown on the liquid Pahm medium and subcultured twice a week. Assays of trichomonad cultures were incubated with one of the following substances: chlorhexidine, metronidazole or P27. For each compound, two different concentrations were applied. After 24h exposition, an antitrichomonal effect of the above substances was assessed, namely a quantity of the surviving trichomonads was microscopically determined and compared with that observed in control cultures. The protozoans showed different susceptibility, depending on kind and concentration of tested compound. In comparison to the control cultures, the reduction in number of surviving protozoans appeared in the assays with metronidazole, chlorhexidine and P27 (to 63%, 52% and 8.7%, respectively). The highest in vitro susceptibility of T. hominis was revealed to a new imidazole derivative P27. The obtained results allow us to suppose that the further studies in this field will be very helpful for explaining the mechanisms of the antitrichomonal activity of the tested compounds as well as for assessment of the influence of various abiotic factors.

Adult↗

[Hydrolytic activity of Trichomonas vaginalis Donné strains].

The hydrolytic activity of Trichomonas vaginalis Donné strains was investigated using API ZYM test (bioMérieux). Among 19 determined hydrolases were found alkaline and acid phosphatases; also in several strains N-acetyl-beta-glucosaminidase and a-glucosidase were demonstrated. The API ZYM test may be used for characterization of enzymatic activity of T. vaginalis strains, but adequate medium is necessary.

Acid Phosphatase↗

The effect of Trichomonas vaginalis and the role of pH on cell culture monolayer viability.

In vitro, Trichomonas vaginalis produces contact-dependent cytotoxicity in which the monolayers are disrupted and monolayer cells die. In contrast, when cell-free filtrates of T. vaginalis culture supernatants are applied to a monolayer, detachment occurs but cell viability is maintained. Growth of T. vaginalis in culture results in a dramatic fall in pH. Both in systems where T. vaginalis grew in physical contact with McCoy cells and where they were physically separate, monolayers still disrupted; but the rate of death of McCoy cells significantly decreased when pH was rigidly controlled. This would indicate that cell death attributed to contact-dependent cytotoxicity is in part due to the drop in pH when T. vaginalis is grown in vitro.

Animals↗

[Molecular cloning and characterization of a RRas homologue gene from Trichomonas vaginalis].

OBJECTIVE: To clone and characterize a RRas-like gene from Trichomonas vaginalis for studying cellular signal transduction pathways in the organism. METHODS: A cDNA clone, which showed homology with RRas proteins of human being, was isolated and sequenced from a cDNA expression library of T. vaginalis. The genomic DNA corresponding to the cDNA sequence was amplified using PCR technique and sequenced. Sequence analysis was performed using BLASTP, RPS-BLAST and Clustal W programs. Phylogenetic tree was constructed and bootstrapped with 1050 replicates using the software MEGA3. RESULTS: The cDNA sequence showed a length of 705 bp with an open reading frame of 615 bp. The deduced amino acid sequence from the open reading frame possesses 205 residuals. Sequencing of the PCR product of genomic DNA revealed that the genomic DNA sequence encompassing the putative 5'-ATG and 3'-stop codons was identical to the cDNA sequence. Sequence analysis demonstrated that this gene was most homologous to the RRas members of Homo sapiens and Mus musculus (both having 51% identity and 70% similarity), and the amino acid sequence contains highly conserved GTP-binding domains and a fully conserved effector domain of human RRas member. Phylogenetic analysis showed that TvRRas clustered with RAS oncoprotein branch and RRAS branch of human. CONCLUSION: The encoding protein probably belongs to a RRas family of T. vaginalis.

Animals↗

[Construction of prokaryotic expression vector for Trichomonas vaginalis silent information regulator 2 and its expression].

Total RNA was isolated from Trichomonas vaginalis and Tv-Sir2-like cDNA was amplified by RT-PCR and cloned into pGEM-T Easy plasmid. A fragment of Tv-Sir2-like cDNA was subcloned into the expression vector pET-41b and expressed in E.coli BL21 with induction of IPTG. The full-length of Tv-Sir2-like cDNA was cloned and sequenced. The prokaryotic expression system of pET-41b/Tv-Sir2-like was constructed. The fusion protein of Tv-Sir2-like was expressed in E. coli BL21, occupying 30% of the total bacterial protein after being induced by IPTG for 5 h. SDS-PAGE analysis showed that the fusion protein was about Mr 59000. The recombinant protein of Tv-Sir2-like is efficiently expressed in E. coli BL21.

Animals↗

Studies on the incidence of Trichomonas vaginalis amongst pregnant women in Jos area of Plateau State, Nigeria.

An investigation was carried out on the incidence of Trichomonas vaginalis amongst pregnant women in Jos area of Plateau State, Nigeria. The pregnant women were made up of 250 women from Jos metropolis and 250 women from Jos rural area. Jos Metropolis women were found to have 37.6% rate of infection while Jos rural women were found to have 24.8% rate of infection. The pH range of the infected women was found to lie between 6-8. Their vaginal discharges were found to be frothy and greenish-yellow with a typical odour. Some had vulval irritations. Some reported that they had slight temperature rises above 37 degrees C in the nights. The infected women either belonged to the working class or were self employed. There was more infection amongst the working class women in the metropolis while the opposite was the case in Jos rural area. Some of the infected women had trichomoniasis alone while some were found to have mixed infection like trichomoniasis and candidosis, trichomoniasis and gonorrhoea or trichomoniasis, candidosis and gonorrhoea. The implications of these results are discussed.

Candidiasis, Vulvovaginal↗

[Metronidazole resistance test in vitro in strains of Trichomonas vaginalis isolated from clinical material. 1. Test conditions].

Metronidazole-resistant clinical isolates of Trichomonas vaginalis display an aerobic type of resistance detectable in vitro only if some oxygen is present. Consequently, adjustment of the optimal concentration of O2 in assay system is of critical importance for dependable determination of minimal lethal concentration (MLC) of the drug. The authors demonstrated that decrease of oxygen content in assay media (from 191.2 nM to 49.5 nM O2.ml-1) resulted in increased susceptibility of the drug-resistant strain IR-78 to metronidazole (from MLC 100 micrograms.ml-1 to MLC 25 micrograms.ml-1). Different MLCs were obtained in media of different reducing capacity. Oxygen dependent variations in MLC were also observed in assay systems containing different volumes of medium. The authors discuss effect of these and other variables in methodology of susceptibility assays in order to define conditions of standard procedure.

Animals↗

[Metronidazole resistance test in vitro in strains of Trichomonas vaginalis isolated from clinical material. 2. Proposal for a standard tests].

The purpose of this paper is to recommend a dependable susceptibility assays for detection of resistance to metronidazole in Trichomonas vaginalis, suitable for routine use in clinical and public health care laboratories. Two different assays, the microtitre plate test based on Meingassner's technique and an alternative tube-assay, were scrutinized and compared using 10 metronidazole-resistant isolates from Czechoslovakia, other European countries and USA and 10 drug-susceptible strains isolated in Czechoslovakia. The minimal lethal concentrations of metronidazole determined for the resistant strains ranged from 25 to 317.5 micrograms.ml-1 while those of drug-susceptible from 3.1-12.5 micrograms.ml-1 metronidazole. Results obtained by both assays were highly reliable and mutually comparable. The authors recommend the microtitre plate test as a method of choice.

Animals↗

[Electron microscopic cytochemistry studies on membrane-bound organelles of Trichomonas hominis].

This paper presents the results of an electron microscopy cytochemistry study on the membrane-bound organelles of Trichomonas hominis. Acid phosphatase (ACP) and cytidine monophosphatase (CMPase) were located in the primary lysosomes, digestive vacuoles, as well as in the parabasal body stacks, being usually in its mature surface sacs. Thiamine pyrophosphatase (TPPase) and nicotinamide adenine dinucleotide phosphatase (NADPase) were found in the saccules, being on the mature surface and in the intermediate saccules of parabasal body stacks, respectively. The reactive products of peroxidase (POase) were seen only in the digestive vacuoles. Cytochrome oxidase (COase) and catalase (Cat) reactions were both negative. It is suggested that T. hominis lacks mitochondrion and microbody but possesses hydrogenosome. The parabasal body, endoplasmic reticulum and hydrogenosome were stained well with the U-Pb-Cu impregnation technique.

Animals↗

[Value of WHO recommendations for the assessment of oral health in patients with diagnosed Trichomonas tenax invasion].

In patients with periodontium lesions infected and not infected with Trichomonas tenax the following indices were analysed: caries index, Russell's index (+periodontal diseases), Greene-Vermillions index (oral hygiene). No statistically significant differences were found between infected and non-infected groups as regards caries and oral hygiene. Russell's index showed considerable differences between the groups of subjects.

Humans↗

Fibronectin- and laminin-mediated endocytic activity in the parasitic protozoa Trichomonas vaginalis and Tritrichomonas foetus.

The pathogenic protozoan Tritrichomonas foetus is able to ingest polystyrene particles with a diameter up to 1.0 micron. Trichomonas vaginalis, however, ingest particles as large as 4.4 microns in diameter. The particles are found within cytoplasmic membrane-bounded vacuoles. Morphometrical analysis showed that T. vaginalis presents a higher endocytic activity than T. foetus. Coating of the polystyrene particles with cationized ferritin increases their binding to the parasite surface but does not interfere with their ingestion. In contrast, coating with laminin significantly increased the uptake of the particles by both parasites while coating with fibronectin potentiates the ingestion of the particles only by T. foetus. These observations suggest the presence of laminin- and fibronectin-binding sites on the surface of trichomonads, an observation which is in agreement with the recent description of a receptor for laminin on the surface of trichomonads.

Animals↗