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Evaluation of the effect of a plant alkaloid (berberine derived from Berberis aristata) on Trichomonas vaginalis in vitro.

Berberine is a quaternary alkaloid derived from the plant Berberis aristata having antibacterial, antiamoebic, antifungal, antihelminthic, leishmanicidal and tuberculostatic properties. The effect of berberine sulphate salt on the growth of Trichomonas vaginalis in vitro was compared to the efficacy of metronidazole as a reference drug. Results showed that berberine sulphate was comparable to metronidazole as regards potency with the advantage of being more safe and possible replacement in metronidazole resistant cases.

Animals↗

A preliminary study on the relationship between Trichomonas vaginalis and cervical cancer in Egyptian women.

The relationship between Trichomonas vaginalis and cancer cervix was investigated by detection of T. vaginalis antibodies, in the sera of 48 invasive cervical cancer patients and 100 random age matched female control, using western immunoblot technique. It was found that antibodies to T. vaginalis were detected in sera of 18.75% (9/48) of cervical cancer patients compared with 5% (5/100) of controls. The increase was evident in age group, 40-49 years (21.05% vs 5%) and of those with squamous cell carcinoma (6/9) and mainly with grade II & III. All the reactive sera of invasive cancer patients reacted strongly with T. vaginalis surface antigen of about (109.9, 86.1, 56.2, 48.2 and 30 Kda). So, there may be an association between T. vaginalis and the risk of cervical cancer, as there was more than 3 fold increase in the prevalence of T. vaginalis antibodies in patients with invasive cervical cancer compared to age matched female controls. This study highlights the importance of clinically detection of T. vaginalis infection, which is one of the group of factors involved in the genesis and progression of cervical cancer. In addition, its treatment would aid in restricting the rising incidence of this dreaded disease.

Adenocarcinoma↗

[Electrophoretic analysis of soluble antigens of a pathogenic isolate of Trichomonas vaginalis].

AIM: To analyze the parasite soluble antigens of a pathogenic isolate of Trichomonas vaginalis. METHODS: The soluble antigens of a pathogenic isolate of T. vaginalis isolated from a patient suffered from trichomonad vaginitis was studied by means of SDS-PAGE followed by densitometric analysis and immunoblotting. RESULTS: A total of 14 distinct protein bands was demonstrated using a 10% resolution gel concentration. Densitometrogram indicated that the soluble antigens with MWs of equal to and below 72 kDa accounted for 89.5% of the total volume with the dominant fractions of 48-51 kDa which accounted for 19.2%. Having MWs over 100 kDa accounted only for 7.4%, while the lowest M. W. fraction demonstrated was found to be at the site of 13 kDa. By immunoblotting, the specific anti-T. vaginalis antibodies raised in mice recognized a broad spectrum of molecular size range of these antigenic fraction. However, no immunological reaction was seen at the 86 kDa site. CONCLUSION: The proteins of MWs 48-51 kDa possess a larger protein content and a higher immunological reactivity.

Animals↗

Effect of the iron chelator deferoxamine on Trichomonas vaginalis in vitro.

The effect of 10 microMol, 15 microMol, 30 microMol, and 60 microMol concentrations of deferoxamine (DFO), a clinically approved iron chelator, was determined on viability and multiplication of Trichomonas vaginalis grown in TYM axenic culture medium at 24 hours interval. DFO killed all T. vaginalis isolates with a minimum lethal concentration of 30 microMol after 48 hours culture incubation with the drug. A potent and persistent inhibitory effect of DFO on the parasite viability and multiplication was recorded throughout the study till its end, in a drug concentration and time exposure-dependent manner. Furthermore, the present work studied the proteinase activity of T. vaginalis grown for 48 hours in DFO inoculated TYM medium, and recorded a significant decrease by all drug concentrations applied in the work. Different possible mechanisms of action of DFO against T. vaginalis and its possible use for treatment of trichomoniasis are discussed.

Animals↗

[Antigen analysis of Trichomonas vaginalis trophozoite by SDS-PAGE and two-dimensional gel electrophoresis].

OBJECTIVE: To analyze soluble antigens of Trichomonas vaginalis. METHODS: Soluble antigens of the parasite from a patient suffering from trichomonad vaginitis were analyzed by SDS-PAGE, immunoblotting and two-dimensional gel electrophoresis. RESULTS: A total of 26 distinct protein bands were demonstrated by using 10% resolution gel. Nine of them were main bands, eight with MWs 15-62 kDa, one with MW 97 kDa. By immunoblotting the specific anti-T. vaginalis antibodies raised in rabbit recognized 24 protein bands with 8 main bands in them. Two-dimensional gel electrophoresis revealed up to 43 individual trichomonad polypeptide spots, among which, 9 were main ones. The pI and MWs of these spots were 3.65-5.84 and 27-->100 kDa respectively. CONCLUSION: Eight protein bands out of 26 soluble antigen bands of the parasite showed high immunogenicity. There were 9 main polypeptide spots in 43 polypeptide spots of the parasite.

Animals↗

[Evaluation of direct microscopic examination, acridine orange staining and culture methods for studies of Trichomonas vaginalis in vaginal discharge specimens].

Trichomonas vaginalis is the causative agent of human trichomoniasis which is a sexually transmitted disease mainly in women. The infection may be asymptomatic or symptomatic such as severe vaginitis and cervicitis. The aim of this study was to compare direct microscopic examination, acridine orange stained examination and culture in Modified Diamond medium, for the detection of T. vaginalis from the vaginal swab samples of 310 patients (age ranges: 17-45 years old) who were complaining from vaginal discharge. Of them 40 (12.9%) samples were found positive with culture, 20 (6.5%) were positive with direct microscopy and 19 (6.1%) were positive with acridine orange staining method. The positive results were obtained in 17 cases by each of the three methods, in 3 cases by direct microscopy and culture, in 2 cases by acridine orange staining and culture, and in 18 cases by culture only. T. vaginalis has been detected in 19.5% of 41 patients with itching, 15.7% of 190 patient with groin pain and 23.2% of 43 patients with cervical erosion, in addition to vaginal discharge, by at least one of the methods. In conditional evaluation, there were no statistically significant differences between T. vaginalis positivity with age groups and the contraceptive methods used. As a result, it was concluded that for the laboratory diagnosis of T. vaginalis, acridine orange staining technique does not have any superiority over direct microscopy. Although direct microscopy is a practical and economical method, it has low sensitivity, so all of the suspected samples which are found negative by this method, should be cultivated for a definite diagnosis.

Acridine Orange↗

[Survival of Trichomonas vaginalis in the environmental media].

The survival of Trichomonas vaginalis outside of the body and from culture media was evaluated under environmental conditions using adsorbent (towels) and non adsorbent (plastic wells) surfaces. After 120 min of exposition under external conditions recovery of parasites from clinical samples showed viable protozoa onto adsorbent and non adsorbent surfaces (5.1% and 30.5%, respectively), as well as for parasites harvested from culture media and assayed onto both surfaces (77.2% and 81.8%, respectively). Parasites from human samples and culture could survive up to 24 h only onto non adsorbent materials (5.1% and 9.1%, respectively). Ability of T. vaginalis to remain viable under external conditions is discussed in relation to its non venereal transmission.

Adolescent↗

Trichomonas urethritis in males.

Trichomonas urethritis in the male should be suspected in all chronic cases of urethritis. The diagnosis is easily established by the hanging-drop method of examining the urethral discharge, or the first-glass urine specimen. Curative treatment is readily accomplished by the use of urethral instillations of Carbarsone suspension using 1 capsule of Carbarsone per ounce of distilled water.

Bacteriology↗

[Detection of Trichomonas vaginalis with direct immunofluorescence assay].

OBJECTIVE: To detect Trichomonas vaginalis (Tv) by direct immunofluorescence assay (DFA) and discuss its clinical significance. METHODS: The parasites at a concentration of 3 x 10(5) cells/L were fixed by acetone on slides which were then blocked by 1% BSA (bovine serum albumin) or 10% BSA or 10% NCS (newborn calf serum) respectively, incubated with different dilution of polyclonal goat anti-Tv IgG (1:20-1:2,560) for different incubation time (15, 30, 45, 60, 90, 120 min). 120 clinical vaginal specimens were examined by direct immunofluorescence assay, the wet mount method and the in vitro cultivation. RESULTS: Blocked by 1% or 10% BSA, incubated at 37 degrees C for 45 min with a titer 1:160 of polyclonal antibody were the optimal conditions for direct immunofluorescence assay. Its sensitivity and specificity were 87.9% and 98.6% respectively in comparison with the in vitro cultivation method. CONCLUSION: Direct immunofluorescence assay is a useful alternative to the wet mount method which shows a lower sensitivity.

Animals↗

Pulmonary trichomoniasis: improved diagnosis by using polymerase chain reaction targeting Trichomonas tenax 18S rRNA gene in sputum specimens.

A polymerase chain reaction (PCR) assay targeting species-specific region in 18 small subunit ribosomal RNA gene of Trichomonas tenax was used to examine sputum specimens in order to diagnose pulmonary trichomoniasis caused by T. tenax. It was compared with wet mount preparation, Giemsa-stained smear, and Kupferberg Trichononas broth culture for detection of T. tenax trophozoites in sputum. The study included 250 individuals; 100 immunocompromised patients with chest complaints (group I) and 100 patients with chronic pulmonary diseases (group II), and 50 healthy individuals as controls (group III). 20 cases among all examined were positive in one or more method giving for pulmonary trichomniasis a total prevalence of 8%; 12 cases (12%) in group I, 8 cases (8%) in group II, and none in group III, with no significant difference between groups I & II. Pulmonary trichomoniasis was prevalent at age ranged between 31 to 50 years, and in total males (10%) than females (5.5%) with no significant difference. Among the 200 examined patients, pulmonary trichomoniasis had a prevalence of 3% by wet mount, 2.5% by Giemsa-stained smear, 7% by culture, compared to 10% by PCR. Culture was used as reference standard. All culture positive specimens were PCR positive showing a product at 0.8 Kb long by agarose gel electrophoresis, and giving a 100% sensitivity. Wet mount, Giemsa-stained smear, and culture had a sensitivity of 43%, 35.7%, and 70%, respectively. No PCR negative specimens were positive by any of the other methods. 6 specimens were culture negative PCR positive and remained PCR positive when retested 3 times. The calculated specificity of PCR was 97%. NO PCR target product was amplified with DNAs of T. vaginalis and various pulmonary pathogens. The results are discussed.

Adult↗

[RAPD analysis on different isolates of Trichomonas vaginalis].

OBJECTIVE: To study genetic polymorphism of DNA on seven isolates of Trichomonas vaginalis. METHODS: The random amplified polymorphic DNA (RAPD) technique was performed to amplify genomic DNA of the seven T. vaginalis isolates, including Beijing 1, Beijing 2, Chengde, Tangshan, Jiujiang 1, Jiujiang 2 and Jiujiang 3. The DNA bands detected were analyzed by clustering analysis with SPSS software. RESULTS: The percentage of genetic similarity among the seven isolates was from 77.4% to 94.7%, showing a close genetic relationship among them. The percentages between the isolates of Beijing 1 and Tangshan, Jiujiang 1 and Jiujiang 2, Beijing 2 and Jiujiang 3 were 89.2%, 92.1% and 94.7% respectively, while that of Jiujiang 1 and Chengde was 77.4%, indicating a lower homology. CONCLUSION: There are a close genetic relationship and certain gene polymorphism among the seven T. vaginalis isolates; geographical origin plays little role to the genetic characteristics.

Animals↗

Surface properties and in vitro cytopathic effect of various strains of Trichomonas vaginalis.

The in vitro cytopathic effect of four strains of Trichomonas vaginalis on cultured epithelial monolayers was analyzed through electrophysiology and electron microscopy. Interaction of trichomonads of two virulent strains (GT-10 and GT-13) with cultured MDCK cell monolayers mounted in Ussing chambers produced a rapid decrease in transepithelial resistance to less than 30% of control values after only 15 min. By 30 min the electrical resistance was practically abolished by the virulent parasites. In contrast, of two attenuated strains of trichomonads (GT-3 and GT-7) analyzed under similar conditions, GT-3 trophozoites required 180 min to reduce transepithelial resistance to 9% of control values, while monolayers in contact with GT-7 parasites still showed 28% of control values at this time of incubation. Sequential scanning electron microscopy confirmed the much faster and widespread cytopathic effect of virulent parasites. In contrast, the slow lytic process produced by attenuated trophozoites was reduced to focal areas of direct contact with epithelial cells. Another difference was found by measurement of the surface charge of the four strains of T. vaginalis by means of cell microelectrophoresis. While the two virulent strains showed a negative surface charge, the two attenuated strains had no detectable surface charge at neutral pH. When parasites were incubated with cationized ferritin and studied with transmission electron microscopy the surface of virulent trichomonads appeared heavily labeled, whereas the surface of attenuated parasites had only sparse and irregular ferritin binding.

Animals↗

Biological variability of Trichomonas vaginalis clinical isolates from symptomatic and asymptomatic patients.

The vaginal specimens of sixty women were cultured using modified thioglycolate medium. Thirteen samples were positive for Trichomonas vaginalis 21.7%. Pathogenicity of these clinical isolates was investigated by intra-peritoneal injection to mice. Additionally, their susceptibility to Metronidazole was evaluated. All the clinical isolates produced infection in mice but marked variations in pathogenicity and susceptibility were observed. Four out of 13 isolates (30.8%) produced mortality in mice indicating higher pathogenicity of the 4 isolates. As to Metronidazole susceptibility, only one isolate was considered resistant and other 12 ones showed different degrees of susceptibility.

Animals↗

[Frequency of Trichomonas vaginalis infection in couples with fertility problems].

One-hundred and seventy two couples which went for the first time to the Infertility Service of the National Institute of Endocrinology from June 1999 to June 2000, were studied to find out the frequency of Trichomonas vaginalis infection in this group, and determine its interaction with a number of clinical and risk variables. The results yielded that 10.5% were positive to the parasite, the prevailing symptom was leukorrea in women and 96.6% of men showed no symptoms. It was highly significant the fact of having a previous pathological history that might be related to infertility and current T. vaginalis infection. This protozoon was frequently associated with Candida sp. and causative agents of bacterial vaginosis in women and Haemophilus influenzae in men. This parasite seems to play an important role as a likely causative agent to be considered in fertility problems.

Adult↗

Molecular characterization of Egyptian Trichomonas vaginalis clinical isolates by HSP70 restriction fragment length polymorphism.

Molecular typing of 20 Egyptian Trichomonas vaginalis clinical isolates was performed using the Restriction Fragment Length Polymorphism (RFLP) analysis employing a probe from the heat-inducible cytoplasmic HSP70 gene family hybridized with EcoR l-digested genomic DNA. In each of the isolates tested 5 to 6 distinct DNA fragments ranging from 2.7 Kb to 7.5 Kb in size were detected. Analysis of 13 isolates from symptomatic and 7 isolates from asymptomatic women revealed 6 distinct RFLP pattern subtypes of T. vaginalis. Eleven isolates (55%) showed the same RFLP pattern, teen of them (90.9%) were from symptomatic patients. T. vaginalis virus (TVV) was present in 7 isolates (35%). Only one isolate was considered resistant to Metronidazole. There were no relations between TVV infection or Metronidazole susceptibility and RFLP subtypes.

Animals↗

Characterization of specific Trichomonas vaginalis target antigens from different isolates by immunoblotting against hyperimmune rabbit serum.

The SDS-PAGE and immunoblot methods were used to identify Trichomonas vaginalis specific target antigen(s). Ten T. vaginalis isolates, cultured on TYM media, were analyzed by Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis (SDS-PAGE), revealed 34 distinct bands. Immunoblotting against a hyperimmune rabbit serum, showed 20 reactive bands ranging from 14-205 kDa. There was isolate to isolate variability among 8 isolates, while 2 isolates (6,7) showed a similar antigenic patterns. Imunoblotting of the isolates showed a total of 20 molecular weight bands, 80% of isolates gave positive immunologic reactions above 100 kDa, while below 100 Kda all the isolates recognized the different molecular weight bands. Out of 20 reacting bands, 5 main bands were detected, 29, 66, 84, 95 & 115 kDa gave positive percent of 60, 60, 60, 90 & 80% among the 10 isolates respectively.

Animals↗

[Detection of Trichomonas vaginalis in men].

An infection with T. vaginalis in male patients is difficult to diagnose in case of latent or asymptomatic trichomoniasis. This infection is caused by the round forms of T. vaginalis, which are significantly more frequent in male patients, than trophozoites. Trichomonas vaginalis round forms called pseudocyst due to their specific biological properties are difficult to observe with actually used techniques. It was shown, that round forms of T. vaginalis may be detected by Giemsa modified technique and modified survival technique employing acridine orange. Therefore routine diagnosis of trichomoniasis included direct examination of the discharge from urethra, cultures and survival technique employing acridine orange. In the problematic cases, especially in male patients with the latent or asymptomatic trichomoniasis, full range of methods should be used to examine specimen collected from the each part of the urinary tract.

Animals↗

[Detection of Mycoplasma hominis in Trichomonas vaginalis by PCR].

Twelve of 28 isolates of Trichomonas vaginalis collected from patients in hospitals in Guangzhou were found naturally infected with Mycoplasma hominis by PCR using specific primers. It suggests that the symbiosis of M. hominis in T. vaginalis also commonly occurs in China.

Animals↗