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Oxidative stress and Trichomonas vaginalis: the effect of hydrogen peroxide in vitro.

The oxidative stress response induced in Trichomonas vaginalis by exposure to various concentrations of hydrogen peroxide (H2O2) was traced by metabolic labeling and monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of trichloroacetic acid-precipitated proteins. Concentrations of H2O2 > 450 microM decreased incorporation of radiolabel into protein and altered protein synthesis resulted in a banding pattern similar to the heat shock protein profile. The proteins produced by oxidative stress included molecules with average molecular masses of 145-165, 92-84, 66, 43-46, and 35-36 kD. Oxidative stress induced changes in T. vaginalis protein synthesis slowly. Full conversion to oxidative stress response occurred within 150-180 min after stress initiation. The oxidative stress responses of three strains freshly initiated from stabilates were compared with the responses of the same strains cultured in vitro for extended periods (culture adapted). Both freshly initiated and culture adapted T. vaginalis of the three strains synthesized the 92-84- and 66-kD heat shock proteins but differed in the synthesis of the 74-75-, 43-, and 35-36-kD molecules. Culture adaptation did not modify the oxidative stress response of the three strains tested.

Animals↗

The mode of action of a novel 18-membered macrolide, virustomycin A (AM-2604 A), on Trichomonas foetus.

The mode of action of virustomycin A, a novel 18-membered macrolide, on Trichomonas foetus was investigated. The antibiotic inhibited the biosynthesis of RNA, DNA and protein in the organism. The inhibition of RNA biosynthesis was the most severe. Virustomycin A repressed the incorporation of [3H]uridine into both acid-soluble and insoluble fractions, whereas actinomycin D inhibited the incorporation of [3H]uridine into acid-insoluble fraction alone. Furthermore, it was found that virustomycin A interfered with nucleotide formation from uridine and adenosine but not with their transport to the cells. On the other hand, the antibiotic did not inhibit the activities of uridine kinase and uracil phosphoribosyltransferase in a cell-free extract from the organism. These data suggest that the antibiotic interferes with the formation of phosphate donor(s) (possibly ATP-forming system) of the organism.

Anti-Bacterial Agents↗

Resolution of six chromosomes of Trichomonas vaginalis and conservation of size and number among isolates.

The electrophoretic karyotype of Trichomonas vaginalis isolates was determined by contour-clamped homogeneous electric field electrophoresis. Six chromosomal bands ranging between 50 kbp and 6 Mbp were reliably resolved by our separation method. Trichomonad chromosomes fell into 3 distinct size classes. The 3 maxichromosomes were approximately 5,700, 4,700, and 3,500 kbp. Two intermediate-sized chromosomes were approximately 1,200 kbp and 1,100 kbp. A minichromosome was approximately 75 kbp. The same size and number of chromosomes were present in 15 T. vaginalis isolates obtained from different geographic regions, reinforcing the idea of a highly conserved karyotype among trichomonal isolates worldwide.

Animals↗

Estimating the prevalence of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and human papillomavirus infection in indigenous women in northern Australia.

OBJECTIVE: To estimate more accurately the age specific prevalence of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and human papillomavirus infection (HPV) in indigenous women living in urban, rural, and remote areas of the "Top End" of the Northern Territory (NT). DESIGN: Analysis of data obtained from two community based studies using self administered tampon specimens tested by polymerase chain reaction for sexually transmitted disease (STD). Data pertaining to the notifiable STDs (N gonorrhoeae and C trachomatis) were obtained from the NT health department. PATIENTS: 1090 indigenous women (age range 12-73 years) were enrolled when they attended local community health centres, family planning clinics, and STD clinics. The majority attended clinics in their home community in the course of "well women's checks" which encourage women to undergo screening for a variety of general medical conditions. RESULTS: The overall prevalence of T vaginalis, C trachomatis, N gonorrhoeae, and HPV was 0.25 (95% CI: 0.22-0.28), 0.11 (0.09-0.13), 0.17 (0.15-0.19), and 0.42 (0.37-0.48) respectively. Of the women found to be infected (excluding HPV), 25.5% had two or more of the above organisms detected. There was a statistically significant increase in the age specific prevalence of T vaginalis but a significant decrease with age for C trachomatis and HPV infection. There was no statistically significant change for N gonorrhoeae with age. CONCLUSIONS: STDs are hyperendemic in this population of indigenous women and the notification data significantly underestimate their prevalence. Distinct patterns of age specific prevalence were demonstrated, highlighting the need to tailor control strategies to specific epidemiological features.

Adolescent↗

In vitro sensitivity of Trichomonas vaginalis to DNA topoisomerase II inhibitors.

Vaginal trichomoniasis is a highly prevalent sexually transmitted disease caused by a microaerophilic protozoan Trichomonas vaginalis. The disease is one of the most common sexually transmitted disease and can augment the predisposition of individuals to human immunodeficiency virus (HIV) infection. Although the disease can be treated with metronidazole and related 5-nitroimidazole, cases of trichomonal vaginitis which are refractory to standard treatment seems to be increasing. Clearly, new antitrichomonad agents are needed and DNA topoisomerase II may acts as a new target for antitrichomonad agents. In this study, in vitro sensitivity of T. vaginalis to DNA topoisomerase II was investigated. Axenic culture of local strain of T. vaginalis was performed. Both eukaryotic and prokaryotic DNA topoisomerase II inhibitors such as ellipticine, amsacrine and fluoroquinolones were tested for effectiveness against T. vaginalis in vitro compared to metronidazole. T. vaginalis was sensitive to metronidazole under aerobic conditions. Minimal inhibitory concentrations (MICs) of eukaryotic DNA topoisomerase II inhibitors, ellipticine and amsacrine, were 6.4 mM and 64 mM, respectively. The MICs of prokaryotic DNA topoisomerase II or DNA gyrase inhibitors; ciprofloxacin, ofloxacin and norfloxacin were 64, 960 and 1,280 mM, respectively. Based on the results, among DNA topoisomerase II inhibitors ellipticine was the most effective drug against T. vaginalis in vitro whereas fluoroquinolones did not show high antitrichomonad activity.

Amsacrine↗

Effects of Mycoplasma spp, Trichomonas fetus, and Campylobacter fetus on ciliary activity of bovine uterine tube organ cultures.

Microscopic observations were made of sections of normal bovine uterine tube (oviduct) and fimbriae maintained in vitro and exposed to agents known to localize in the bovine genital tract, i.e., mycoplasma, Trichomonas fetus, and Campylobacter fetus (Vibrio fetus). Only C fetus stopped ciliary activity. Scanning electron microscopy (SEM) revealed loss of most cilia. Sterile filtrate of C fetus culture had no effect.

Animals↗

Trichomonas vaginalis in vaginal smears of women using intrauterine contraceptive device.

To detect the association between the use of an intrauterine contraceptive device (IUCD) and the presence of Trichomonas vaginalis (Tv), vaginal smears of 1110 women were examined cytologically. Among 45 IUCD users, 10 women were positive for Tv (22.22%). Forty six of 1065 non-users were also positive for Tv (4.32%). There was a significant correlation between the use of IUCD and the presence of Tv (p < 0.05). The data also indicated that the prolonged use (> or = 3 years) of copper-IUCD may promote the growth of Tv in the vaginal mucosa. This result will be a clue to diagnose asymptomatic patients for Tv.

Adult↗

Ultrastructural cytochemical observations of the cell coat of two Trichomonas vaginalis isolates with different degree of virulence.

The cell coat of two Trichomonas vaginalis isolates with different degree of virulence isolated in Mexico from symptomatic women was studied by cytochemical assays. The use of carbohydrate cell surface markers allowed us to visualize greater electron-dense deposits in the highly virulent T. vaginalis isolate than in the less virulent one. On the contrary, parasites treated with concanavalin A showed a heavy uniform electron-dense deposit on the cell surface that was similar in both isolates. When parasites were treated with cationized ferritin the amount of bounded particles on the cell surface was greater in the higher virulent isolate.

Animals↗

[Comparative effect of different media in in vitro cultivation of Trichomonas vaginalis].

OBJECTIVE: To investigate the optimal condition for in vitro cultivation of Trichomonas vaginalis for obtaining a better harvest of T. vaginalis. METHODS: An isolate of T. vaginalis from clinical specimens was cultivated in three different media with initial inoculation of 9.0 x 10(4)/ml under pH 5.6. RESULTS: There was distinct difference after 96 h incubation in the cumulative harvest of T. vaginalis. The highest harvest was received in cysteine/liver/peptone/maltose medium, followed by the liver/peptone/maltose medium and soybean/liver/peptone/maltose medium. CONCLUSION: The cysteine/liver/peptone/maltose medium may be a suitable environment for in vitro multiplication of T. vaginalis.

Animals↗

[Studies on ultrastructure and histochemistry of Trichomonas vaginalis adhering to vaginal mucosa of rats].

OBJECTIVE: To study interaction between Trichomonas vaginalis and epithelium of genital tract of host as well as the pathogenesis of T. vaginalis. METHODS: Immunohistochemical technique was used to observe the adhering process of T. vaginalis to vaginal mucosa in rats by transmission and scanning microscopy. RESULTS: T. vaginalis were shown to be PAS positive and clusters of T. vaginalis were found to adhere to columnar epithelium rich in mucopolysaccharide on the surface of vaginal mucosa as viewed in sections of the middle and upper parts of the orgen. T. vaginalis was positive for cathepsin. And the membrane of epithelial cells was often damaged by the released hydrolase. The parasite was also positive for actin; the microfilament bundles were arranged in reticular form in ameboid T. vaginalis. The latter would penetrate between epithelial cells, and its filiform pseudopodia would invade the interspace of microvilli of the epithelium, to encircle and gradually phagocytize microvilli. Digitiform pseudopodia would insert between epithelial cells and encircle part of them. A few T. vaginalis were found to adhere to the keratinized epithelium between mucosal folds as shown in the sections of the lower part of vagina. CONCLUSION: T. vaginalis is inclined to parasitise vaginal fornix because the superficial epithelial cells there are rich in mucinogen granules and abundant microvilli exist. After adhering, T. vaginalis releases hydrolase to digest and phagocytize epithelium which may directly damage the epithelium of the genital canal. Moreover, T. vaginalis would take in mucopolysaccharide to affect the normal clearance process of vagina, resulting in inflammation of parasitized tissue. The cytoskeleton, cell coat, polymorphism of pseudopodia and lysosome of T. vaginalis play an important role in the courses of movement, adhesion, encirclement, phagocytosis and digestion.

Actins↗

Ultracytochemical studies on Trichomonas hominis.

The results of ultracytochemical studies on Trichomonas hominis showed that ACPase and CMPase were mainly located in the mature face sacs of the primary lysosomes, digestive vacuoles, as well as in the parabasal body. TPPase and NADPase were found in the saccules at the mature face and the intermediate saccules of parabasal body respectively. This study revealed that T. hominis had well-developed parabasal bodies. Negative COase and catalase reactions indicated that T. hominis lacked both mitochondria and microbody. Hydrogenosome was stained well with the Ur-Pb-Cu impregnation technique.

Acid Phosphatase↗

Cell surface saccharide differences in drug-susceptible and drug-resistant strains of Trichomonas vaginalis.

Surface carbohydrates of drug-resistant and drug-susceptible strains of Trichomonas vaginalis were analysed using lectins. The presence of D-GalNAc, D-Gal and mannose-like residues was detected in T. vaginalis. Marked differences in exposed surface carbohydrates were documented, e.g. wheat germ agglutinin (WGA) selectively agglutinated the drug-susceptible strain whereas drug-resistant parasites reacted preferentially with concanavalin A (Con A). In drug-resistant, but not in drug-susceptible strains, trypsinization induced the appearance of soybean agglutinin. Binding studies using fluorescein-labelled WGA and Con A essentially confirmed the agglutination experiments. Both the intense cell agglutination and the fluorescent WGA-binding displayed by a drug-susceptible strain, were completely nullified by neuraminidase treatment, suggesting the presence of an exposed sialic acid moiety on the T. vaginalis surface.

Agglutination Tests↗

The in vitro effect of Assafoetida on Trichomonas vaginalis.

Assafoetida is an oleo-gum resin obtained from the roots and stems of many Ferula species such as F. foetida Regel, F. asafoetida Linn, F. alliacea Boiss. and F. narthex Boiss, family Umbelliferae (carrot family), plants growing indigenously in Iran, Afghanistan and north of India. Assafoetida is commonly used as a flavoring agent in food and as a traditional medicine for many diseases in many parts of the world. In this work, the effect of Assafoetida on growth of Trichomonas vaginalis in vitro was studied and compared to metronidazole as a reference drug. It showed that Assafoetida had a potent antiparasitic effect on T. vaginalis compared to metronidazole, thus, worth further investigation to study its applicability in treatment of parasitic infections.

Animals↗

[Electron microscopic study on Trichomonas vaginalis adhering to and phagocytizing male genitourinary epithelial cells].

OBJECTIVE: To observe Trichomonas vaginalis (T. vaginalis) adhering to and phagocytizing male genitourinary epithelial cells in order to study the pathogenetic mechanism of male trichomoniasis. METHODS: Cultured T. vaginalis bodies were incubated with male genitourinary epithelial cells, and then the ultrastructure was observed with transmission electron microscopy. RESULTS: T. vaginalis adhered to epithelial cells like amoeba, and formed pseudopodium or surface invagination surrounding or nibbling other parts of the epithelial cytoplasm. CONCLUSIONS: T. vaginalis has the speciality of adhering to and phagocytizing to male genitourinary epithelial cells. Genitourinary epithelial cells may be injured directly by the phagocytosis of T. vaginalis. Attention has to be paid to the correlation.

Animals↗

Ultrastructural and immunohistochemical studies on Trichomonas vaginalis adhering to and phagocytizing genitourinary epithelial cells.

BACKGROUND: Trichomonas vaginalis (T. vaginalis) belongs to a common sexually transmitted disease pathogen causing genitourinary trichomoniasis in both sexes. We investigated the pathogenetic mechanism of genitourinary trichomoniasis. METHODS: Cultured T. vaginalis bodies were injected into the vaginas of rats, or incubated with genitourinary epithelial cells of female subjects, male subjects, and sperm. The ultrastructural and microscopic changes were observed via transmission and scanning electron microscopy and through microscopic histochemistry. RESULTS: Groups of T. vaginalis adhered to PAS positive columnar cells at the surface of stratified epithelium in the middle and upper portions of the vaginas. They also traversed under these cells. The parasites were shown to be PAS, cathepsin D, and actin positive, and they could release hydrolase into the cytoplasm of adhered epithelial cells. In the amebiform T. vaginalis, microfilaments were arranged into reticular formation. Similar phenomena were found during the interaction of T. vaginalis with host cells, both in vitro and in vivo. Usually several protozoa adhered to an epithelial cell and formed polymorphic pseudopodia or surface invaginations to surround and phagocytize the microvilli or other parts of the epithelial cytoplasm. Adhesion and phagocytosis of sperm by the protozoa occurred at 15 - 30 minutes of incubation. Digestion of sperm was found at 45 - 75 minutes and was complete at 90 - 105 minutes. CONCLUSIONS: T. vaginalis tends to parasitize at the fornix of the vagina, because this is the site where columnar cells are rich in mucinogen granules and their microvilli are helpful for adhesion and nibbling. T. vaginalis possesses some invading and attacking abilities. Shape change, canalization, encystation, phagocytosis, digestion, the cell coat, cytoskeleton, and lysosome all play important roles in the process of adhesion. They have two methods of phagocytosis: nibbling and ingestion. Genitourinary epithelium may be injured directly by the digestive action of hydrolases, phagocytosis, and the mechanical action of pseudopodia.

Animals↗