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1.
Diagn Cytopathol ; 49(9): 1052-1055, 2021 Sep.
Article En | MEDLINE | ID: mdl-34291890

OBJECTIVE: The parasite Trichomonas vaginalis (T. vaginalis) causes one of the most common non-viral sexually transmitted infections in humans. T. vaginalis is notorious for its inconspicuous appearance in vaginal smears. It can be missed under the microscope. METHOD: In the present study, we investigate the immunoreactivity of T. vaginalis to smooth muscle actin (SMA) in the vaginal smear. RESULT: T. vaginalis trophozoite and pseduocyst are immunoreactive for SMA in all of the study group cases (n = 21) and in none of the control group cases (n = 21). Thus, SMA immunostain is a sensitive method for the demonstration of T. vaginalis. Moreover, the protozoan attains a conspicuous and unique appearance. By SMA immunohistochemical stain, the apperance of T. vaginalis floated freely or located in the cytoplasm of the epithelial cells is easily identified. CONCLUSION: We recommend performing SMA immunostain in every vaginal smear with clinical or pathologic suspicion of trichomoniasis.


Actins/immunology , Protozoan Proteins/immunology , Trichomonas Infections/diagnosis , Trichomonas vaginalis/immunology , Epithelial Cells/parasitology , Female , Humans , Immunohistochemistry/methods , Molecular Diagnostic Techniques/methods , Trichomonas Infections/parasitology , Trichomonas vaginalis/isolation & purification , Trichomonas vaginalis/pathogenicity , Vaginal Smears/methods
2.
PLoS One ; 15(12): e0242227, 2020.
Article En | MEDLINE | ID: mdl-33264307

BACKGROUND: Information about the use of flow cytometry in the diagnosis of male urethritis is scarce. The current study aims to evaluate the performance of flow cytometry on first-voided urine in males with infectious urethritis (Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium and Trichomonas vaginalis). METHODS: Male patients of the Andrology Centre (Tartu University Hospital, Estonia) were recruited during the period March 2015 -January 2018. Cases included 306 patients with infectious urethritis caused by Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium and/or Trichomonas vaginalis. The control group consisted of 192 patients without uro-genital complaints, negative tests for C. trachomatis, N. gonorrhoeae, M. genitalium and T. vaginalis from first-voided urine and no inflammation in first-voided urine, mid-stream urine and urine after prostate massage. C. trachomatis, N. gonorrhoeae, M. genitalium and T. vaginalis were detected from first-voided urine using polymerase chain reaction (PCR) method. First-voided urine was analysed using urine particle analyzer Sysmex UF-500i. RESULTS: The most prevalent infection was chlamydia (64.1%), followed by Mycoplasma genitalium (20.9%), gonorrhoea (7.8%) and trichomoniasis (1.6%). Gonorrhoea caused the highest flow-cytometric leucocyte/bacteria count, followed by chlamydia and Mycoplasma genitalium. Trichomonas vaginalis showed nearly absent inflammation in first-voided urine. Using an empiric flow-cytometry diagnostic threshold for urethritis in first-voided urine (leucocytes ≥ 15/µl and bacteria ≥ 20/µl) the total calculated sensitivity was over 90%. However, when applying such criteria for deciding whether to perform first-voided urine PCR for C. trachomatis, N. gonorrhoeae, M. genitalium and T. vaginalis or not, we could miss 23 cases with infectious urethritis that makes up 7,5% of all proven cases. CONCLUSIONS: Flow cytometry of first-voided urine can be considered as a rapid and objective screening method in case of suspected male infectious urethritis.


Heterosexuality , Inflammation/urine , Sexually Transmitted Diseases/urine , Urethritis/urine , Adolescent , Adult , Body Fluids/microbiology , Chlamydia Infections/complications , Chlamydia Infections/microbiology , Chlamydia Infections/urine , Chlamydia trachomatis/pathogenicity , Estonia/epidemiology , Gonorrhea/complications , Gonorrhea/microbiology , Gonorrhea/urine , Humans , Inflammation/etiology , Inflammation/pathology , Male , Middle Aged , Mycoplasma Infections/complications , Mycoplasma Infections/microbiology , Mycoplasma Infections/urine , Mycoplasma genitalium/pathogenicity , Neisseria gonorrhoeae/pathogenicity , Sexually Transmitted Diseases/complications , Sexually Transmitted Diseases/microbiology , Trichomonas Infections/complications , Trichomonas Infections/microbiology , Trichomonas Infections/urine , Trichomonas vaginalis/pathogenicity , Urethritis/etiology , Urethritis/microbiology , Urethritis/pathology , Young Adult
3.
Pathol Res Pract ; 216(12): 153234, 2020 Dec.
Article En | MEDLINE | ID: mdl-33075741

Gardnerella vaginalis (GV) and Trichomonas vaginalis (TV) infections have been proposed as risk factors for persistence or progression of low-grade precancerous cervical lesions (CIN1/L-SIL). However, their role is still undefined. We aimed to assess if GV and TV infections affect the risk of persistence/progression of CIN1/L-SIL. A retrospective cohort study was performed to assess the risk of CIN1/L-SIL persistence or progression, persistence alone and progression alone in patients with GV and/or TV infections (GV + and/or TV+), only GV (GV+), only TV (TV+), or GV and TV coinfections compared to patients without these infections. Relative risk (RR) with 95 % confidence intervals (CI) was adopted (significant p-value>0.05). Two hundred and seventy patients were included. RR for CIN1/L-SIL persistence or progression was 1.63 in GV + and/or TV+ (p = 0.02), 1.99 in GV+ (p = 0.0008), 0.25 in TV+ (p = 0.32), 1.78 in coinfection (p = 0.26). RR for persistence was 1.55 in GV + and/or TV+ (p = 0.1), 2.179 in GV+ (p = 0.0013), 0.32 in TV+ (p = 0.41), 0.45 in coinfection (p = 0.55). RR for progression was 1.92 in GV + and/or TV+ (p = 0.22), 1.34 in GV+ (p = 0.68), 1.16 in TV+ (p = 0.91), 8.39 in coinfection (p = 0.0002). In conclusion, GV infection may be a risk factor for CIN1/L-SIL persistence. TV infection alone does not significantly affect the risk of persistence or progression of such lesions, while it may greatly increase the risk of progression when associated with GV infection.


Gardnerella vaginalis/pathogenicity , Gram-Positive Bacterial Infections/microbiology , Trichomonas Vaginitis/parasitology , Trichomonas vaginalis/pathogenicity , Uterine Cervical Dysplasia , Uterine Cervical Neoplasms , Vaginosis, Bacterial/microbiology , Adult , Disease Progression , Female , Gram-Positive Bacterial Infections/diagnosis , Humans , Middle Aged , Neoplasm Grading , Retrospective Studies , Risk Assessment , Risk Factors , Trichomonas Vaginitis/diagnosis , Uterine Cervical Neoplasms/microbiology , Uterine Cervical Neoplasms/parasitology , Uterine Cervical Neoplasms/pathology , Vaginosis, Bacterial/diagnosis , Uterine Cervical Dysplasia/microbiology , Uterine Cervical Dysplasia/parasitology , Uterine Cervical Dysplasia/pathology
4.
Cell Microbiol ; 22(11): e13257, 2020 11.
Article En | MEDLINE | ID: mdl-32858768

Extracellular vesicles (EVs) have emerged as a ubiquitous mechanism for transferring information between cells and organisms across all three kingdoms of life. Parasitic unicellular eukaryotes use EVs as vehicles for intercellular communication and host manipulation. Pathogenic protozoans are able to modulate the immune system of the host and establish infection by transferring a wide range of molecules contained in different types of EVs. In addition to effects on the host, EVs are able to transfer virulence factors, drug-resistance genes and differentiation factors between parasites. In this review we cover the current knowledge on EVs from anaerobic or microaerophilic extracellular protozoan parasites, including Trichomonas vaginalis, Tritrichomonas foetus, Giardia intestinalis and Entamoeba histolytica, with a focus on their potential role in the process of infection. The role of EVs in host: parasite communication adds a new level of complexity to our understanding of parasite biology, and may be a key to understand the complexity behind their mechanism of pathogenesis.


Entamoeba histolytica/physiology , Extracellular Vesicles/metabolism , Giardia lamblia/physiology , Host-Parasite Interactions , Trichomonas/physiology , Anaerobiosis , Animals , Entamoeba histolytica/pathogenicity , Entamoebiasis , Giardia lamblia/pathogenicity , Giardiasis/parasitology , Humans , Protozoan Proteins/metabolism , Trichomonas/pathogenicity , Trichomonas Infections/parasitology , Trichomonas vaginalis/pathogenicity , Trichomonas vaginalis/physiology , Tritrichomonas foetus/pathogenicity , Tritrichomonas foetus/physiology
5.
Parasitol Res ; 119(8): 2649-2657, 2020 Aug.
Article En | MEDLINE | ID: mdl-32583161

Trichomonas vaginalis is the most common nonviral sexually transmitted infection. According to the 2019 WHO cancer report, cervical cancer is the fourth most frequent cancer in women. However, previous research, which has not included a large-scale study to date, has revealed that Trichomonas vaginalis increases cervical cancer risk. In this study, we investigated a group of Asian females in Taiwan to determine the association between trichomoniasis and the risk of developing cervical lesions, including cancer, neoplasm, and dysplasia. We conducted a nested case-control study by using the National Health Insurance (NHI) program database in Taiwan. The International Classification of Diseases, 9th Revision classifications (ICD-9-CM) was used to categorize all of the medical conditions for each patient in the case and control groups. The adjusted odds ratio (AOR) and 95% confidence interval (CI) for the association between trichomoniasis and cervical lesions were estimated using multivariable conditional logistic regression to adjust for all comorbidities and variables. In total, 54,003 individuals were enrolled in the case group and 216,012 were enrolled in the control group. Trichomonas vaginalis exposure had a significant association with cervical lesions (AOR 2.656, 95% CI = 1.411-5.353, p = 0.003), especially cervical cancer (AOR 3.684, 95% CI = 1.622-6.094, p = 0.001). In patients with both trichomoniasis and depression, the relative risk increased 7.480-fold compared to those without trichomoniasis or depression. In conclusion, female patients with Trichomonas vaginalis exposure had a significantly higher risk of developing cervical lesions (especially cervical cancer) than those without exposure.


Trichomonas Infections/complications , Trichomonas vaginalis/pathogenicity , Uterine Cervical Diseases/pathology , Uterine Cervical Diseases/parasitology , Adult , Case-Control Studies , Depression/complications , Depression/epidemiology , Female , Humans , Middle Aged , Odds Ratio , Taiwan/epidemiology , Trichomonas Infections/epidemiology , Uterine Cervical Diseases/epidemiology , Uterine Cervical Diseases/psychology , Uterine Cervical Neoplasms/epidemiology , Uterine Cervical Neoplasms/parasitology , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/psychology
6.
Sex Transm Dis ; 47(5): 332-337, 2020 05.
Article En | MEDLINE | ID: mdl-32149959

BACKGROUND: Trichomonas vaginalis is a common treatable sexually transmitted infection among older women. Persistent T. vaginalis infection after treatment is common among women with human immunodeficiency virus (HIV). We sought to determine if HIV-negative women were as likely as women with HIV to have persistent T. vaginalis infection. METHODS: We performed a retrospective cohort study of women 45 years or older with T. vaginalis infection. We evaluated differences in persistent T. vaginalis infection according to HIV status using χ analysis. We performed regression analyses to describe factors associated with persistent and recurrent infection in older women. RESULTS: Over a 10-year study period, we identified 282 women with T. vaginalis, 46 with HIV. Most women (240, 86%) were treated in accordance with 2015 Centers for Disease Control and Prevention Sexually Transmitted Diseases treatment guidelines. Half of the women (144, 53%) had a repeat T. vaginalis test 90 to 365 days after treatment, and one third had persistent infection (39/125, 31%). Persistent infection was similar between women with HIV and HIV-negative women treated according to Centers for Disease Control recommendations (17% vs 33%, P = 0.3). When adjusting for age and incidental diagnosis, tobacco use was associated with an increased risk of more than 1 or recurrent T. vaginalis infection during the study period (adjusted odds ratio, 2.8; 95% confidence interval, 1.5-4.9). CONCLUSIONS: The HIV status did not affect persistent T. vaginalis infection in women 45 years or older. Given over one third of women have a positive test within a year after the recommended treatment, our data support repeat testing in women 45 years and older treated for T. vaginalis.


HIV Infections/complications , Trichomonas Vaginitis/diagnosis , Trichomonas Vaginitis/drug therapy , Trichomonas vaginalis/drug effects , Trichomonas vaginalis/pathogenicity , Aged , Aged, 80 and over , Cohort Studies , Female , Follow-Up Studies , HIV Infections/diagnosis , HIV Infections/drug therapy , HIV Seronegativity , Humans , Middle Aged , Prevalence , Retrospective Studies , Risk Factors , Trichomonas Vaginitis/epidemiology , Trichomonas vaginalis/isolation & purification
7.
Molecules ; 25(4)2020 Feb 12.
Article En | MEDLINE | ID: mdl-32059495

We prepared a series of 10 carbamates derivatives based on two common antiprotozoal drugs: metronidazole (1-5) and secnidazole (6-10). The compounds were tested in vitro against a set of two amitochondriate protozoa: Giardia duodenalis and Trichomonas vaginalis. Compounds 1-10 showed strong antiprotozoal activities, with potency values in the low micromolar-to-nanomolar range, being more active than their parent drugs. Metronidazole carbamate (1) was the most active of the series, with nanomolar activities against G. duodenalis (IC50 = 460 nM) and T. vaginalis (IC50 = 60 nM). The potency of compound 1 was 10 times greater than that of metronidazole against both parasites. None of compounds showed in vitro cytotoxicity against VERO cells tested at 100 µM. Molecular dynamics of compounds 1-10, secnidazole, and metronidazole onto the ligand binding site of pyruvate-ferredoxin oxidoreductase of T. vaginalis and the modeled -tubulin of G. duodenalis revealed putative molecular interactions with key residues in the binding site of both proteins implicated in the mode of action of the parent drugs.


Antiprotozoal Agents/pharmacology , Carbamates/chemistry , Metronidazole/analogs & derivatives , Metronidazole/chemistry , Antiprotozoal Agents/chemical synthesis , Antiprotozoal Agents/chemistry , Carbamates/chemical synthesis , Carbamates/pharmacology , Giardia lamblia/drug effects , Giardia lamblia/pathogenicity , Giardiasis/drug therapy , Giardiasis/parasitology , Metronidazole/chemical synthesis , Metronidazole/pharmacology , Trichomonas Infections/drug therapy , Trichomonas Infections/parasitology , Trichomonas vaginalis/drug effects , Trichomonas vaginalis/pathogenicity
8.
Microb Pathog ; 137: 103752, 2019 Dec.
Article En | MEDLINE | ID: mdl-31539586

We performed a systematic review and meta-analysis to reveal the association between Trichomonas vaginalis (TV) infection and the risk of prostate cancer (PCa) development. Systematic searching (PubMed/Medline, Scopus, Embase, Web of Science, Cinhal, Wiley, Cochrane, Psychoinfo, ProQuest and Google Scholar) was done, up to March 2018 for case-control studies. Random effects model was applied to define odds ratios and their 95% confidence intervals. In total, 6 enteries were included in our meta-analysis, comprising 5590 individuals (2677 PCa cases and 2913 control individuals) examined for trichomoniasis, with a total positivity of 469 (17.51%) and 482 (16.54%) individuals, respectively. Totally, such association was documented in three countries, including USA (4 studies), Kuwait (one study) and Taiwan (one study). Based on pooled estimations, however a 1.17-time increase of PCa was observed among individuals with a previous exposure of TV, it was not statistically significant [OR = 1.17 (95% CI: 1.01 to 1.36)]. Egger's regression test demonstrated no publication bias among studies. Also, year of publication for included records was not significantly correlated to the relationship between trichomoniasis and PCa. Any further inferences should be based on future investigations for better understanding this relationship and shedding light on the cryptic pathogenesis of TV in PCa patients.


Prostatic Neoplasms/complications , Trichomonas Infections/complications , Databases, Factual , Humans , Male , Odds Ratio , Prostatic Neoplasms/epidemiology , Risk Factors , Trichomonas Infections/epidemiology , Trichomonas vaginalis/pathogenicity
9.
Clin Lab ; 65(7)2019 Jul 01.
Article En | MEDLINE | ID: mdl-31307183

BACKGROUND: Prostate cancer is considered the most prevalent cancer among men. Recent studies suggest that sex-ually transmissible infections (STIs) may be related to prostate carcinogenesis. Therefore, the aim of this study was to investigate whether STI pathogens (Atopobium vaginae (ATO), Neisseria gonorrhoeae (NG), Chlamydia tra-chomatis (CT), Treponema pallidum (TP), Ureaplasma urealyticum (UU), Gardnerella vaginalis (GV), Herpes Sim-plex Virus (HSV), Cytomegalovirus (CMV), Human herpesvirus (HHV), Human papillomavirus (HPV), and Tricho-monas vaginalis (TV)) presence in prostate tissues are associated with the risk of prostate cancer. METHODS: Paraffin-embedded prostate tissues obtained from patients with hyperplasia and prostate cancer were extracted. Determination of infectious microorganisms of interest was done by quantitative TaqMan real-time PCR assay. RESULTS: STI DNA was detected in 53/243 (21.8%) of the prostate tissues samples (ATO 3.7%, UU 2.88%, GV 2.46%, HSV-2 2.05%, CT 2.05%, CMV 1.64%, NG 1.64%, TP 1.64%, HHV-8 1.23%, HPV 1.23%, and TV 1.23%.) The statistical analysis revealed significant correlation between prevalence of Gardnerella vaginalis (GV) and Herpes Simplex Virus (HSV-2) between hyperplasia and cancerous groups (p = 0.02), respectively. CONCLUSIONS: No statistically significant difference was observed in the prevalence of most candidate infectious or-ganisms between hyperplasia and cancerous groups except for GV and HSV-2. It appears that inflammation in the prostate gland is more associated with prostate hyperplasia than prostate cancer. According to the role of in-fectious microorganisms in induction of chronic inflammation, we cannot exclude the importance of these patho-gens in progression of cancer. More studies are required to explore the associations of cancer with different infec-tious organisms.


Prostatic Hyperplasia/diagnosis , Prostatic Neoplasms/diagnosis , Real-Time Polymerase Chain Reaction/methods , Sexually Transmitted Diseases/complications , Aged , Chlamydia trachomatis/genetics , Chlamydia trachomatis/pathogenicity , Host-Pathogen Interactions , Humans , Male , Middle Aged , Neisseria gonorrhoeae/genetics , Neisseria gonorrhoeae/pathogenicity , Papillomaviridae/genetics , Papillomaviridae/pathogenicity , Prostate/microbiology , Prostate/parasitology , Prostate/virology , Prostatic Hyperplasia/complications , Prostatic Neoplasms/complications , Trichomonas vaginalis/genetics , Trichomonas vaginalis/pathogenicity , Virulence/genetics
10.
J Infect Chemother ; 25(12): 955-964, 2019 Dec.
Article En | MEDLINE | ID: mdl-31189504

Trichomonas vaginalis (T. vaginalis) is a common sexually transmitted infection, affecting the urogenital tract. Trichomoniasis is customarily treated with metronidazole (MTZ). MTZ is known to cause undesirable side effects and there is several reports on MTZ resistant T. vaginalis. Thus, the present study aimed to in-vitro evaluate the activity of DNA minor groove binder drug ''Netropsin dihydrochloride'' against metronidazole-sensitive T. vaginalis isolates (G and U isolates) and resistant T. vaginalis isolate (ATCC50138) (R isolate). Netropsin was tested at concentrations ranging from 3.5 to 200 µg/ml. It showed effectiveness against all isolates with MLC of 12.5 µg/ml for G and U isolates and of 25 µg/ml for R isolate. Cytotoxicity assay of isolates exposed to the respective MLC of netropsin for 42 h showed a highly significant reduction in the death percentage of MCDK cell line as compared to the effect elicited by drug free controls. The hemolytic activity was evaluated by hemolytic assay and by monitoring the interaction of T. vaginalis isolates with human erythrocytes by inverted microscopy and scanning electron microscopy. The hemolytic assay showed (0%) hemolysis of RBCs incubated with T. vaginalis isolates treated with the corresponding MLC of netropsin for 24 h. Scanning electron microscopy revealed cytoskeletal deformities of netropsin treated isolates. Taken together, these observations suggest that netropsin is a promising therapy for T. vaginalis infection affecting its viability, virulence, cytopathogenic and hemolytic activity with a mechanism of action that might overcome T. vaginalis resistance to metronidazole.


Anti-Bacterial Agents/pharmacology , Netropsin/pharmacology , Trichomonas Vaginitis/drug therapy , Trichomonas vaginalis/drug effects , Animals , Anti-Bacterial Agents/therapeutic use , Dogs , Drug Resistance , Female , Hemolysis/immunology , Humans , Madin Darby Canine Kidney Cells , Metronidazole/pharmacology , Metronidazole/therapeutic use , Netropsin/therapeutic use , Parasitic Sensitivity Tests , Trichomonas Vaginitis/parasitology , Trichomonas vaginalis/immunology , Trichomonas vaginalis/isolation & purification , Trichomonas vaginalis/pathogenicity , Trophozoites/drug effects , Trophozoites/immunology , Vagina/parasitology
11.
mBio ; 10(3)2019 05 14.
Article En | MEDLINE | ID: mdl-31088924

Trichomonas vaginalis, a prevalent sexually transmitted parasite, adheres to and induces cytolysis of human mucosal epithelial cells. We have characterized a hypothetical protein, TVAG_393390, with predicted tertiary structure similar to that of mammalian cadherin proteins involved in cell-cell adherence. TVAG_393390, renamed cadherin-like protein (CLP), contains a calcium-binding site at a position conserved in cadherins. CLP is surface localized, and its mRNA and protein levels are significantly upregulated upon parasite adherence to host cells. To test the roles of CLP and its calcium-binding dependency during host cell adherence, we first demonstrated that wild-type CLP (CLP) binds calcium with a high affinity, whereas the calcium-binding site mutant protein (CLP-mut) does not. CLP and CLP-mut constructs were then used to overexpress these proteins in T. vaginalis Parasites overexpressing CLP have ∼3.5-fold greater adherence to host cells than wild-type parasites, and this increased adherence is ablated by mutating the calcium-binding site. Additionally, competition with recombinant CLP decreased parasite binding to host cells. We also found that overexpression of CLP induced parasite aggregation which was further enhanced in the presence of calcium, whereas CLP-mut overexpression did not affect aggregation. Lastly, parasites overexpressing wild-type CLP induced killing of host cells ∼2.35-fold, whereas parasites overexpressing CLP-mut did not have this effect. These analyses describe the first parasitic CLP and demonstrate a role for this protein in mediating parasite-parasite and host-parasite interactions. T. vaginalis CLP may represent convergent evolution of a parasite protein that is functionally similar to the mammalian cell adhesion protein cadherin, which contributes to parasite pathogenesis.IMPORTANCE The adherence of pathogens to host cells is critical for colonization of the host and establishing infection. Here we identify a protein with no known function that is more abundant on the surface of parasites that are better at binding host cells. To interrogate a predicted function of this protein, we utilized bioinformatic protein prediction programs which allowed us to uncover the first cadherin-like protein (CLP) found in a parasite. Cadherin proteins are conserved metazoan proteins with central roles in cell-cell adhesion, development, and tissue structure maintenance. Functional characterization of this CLP from the unicellular parasite Trichomonas vaginalis demonstrated that the protein mediates both parasite-parasite and parasite-host adherence, which leads to an enhanced killing of host cells by T. vaginalis Our findings demonstrate the presence of CLPs in unicellular pathogens and identify a new host cell binding protein family in a human-infective parasite.


Cadherins/genetics , Epithelial Cells/metabolism , Protozoan Proteins/metabolism , Trichomonas vaginalis/pathogenicity , Cadherins/metabolism , Calcium/metabolism , Cell Adhesion , Cell Line , Epithelial Cells/parasitology , Female , Humans , Mucous Membrane/cytology , Protein Domains , Protozoan Proteins/genetics , Transcriptional Activation , Up-Regulation
12.
Infect Immun ; 87(5)2019 03.
Article En | MEDLINE | ID: mdl-30858343

The human protozoan Trichomonas vaginalis is the causative agent of trichomoniasis, a prevalent sexually transmitted infection, which is accompanied by a species-diversified vaginal microbiota named community state type IV (CST-IV). Coincidently, CST-IV includes species associated with bacterial vaginosis (e.g. Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia). Both diseases are linked to the transmission of human immunodeficiency virus (HIV) and preterm birth, which complications are likely to result from the disruption of the cervicovaginal epithelial barrier. Here, we show that paracellular permeability of fluorescein isothiocyanate (FITC)-dextran through a monolayer of human ectocervical cells (hECs) is increased as a consequence of the activity of T. vaginalis and the aforementioned species of CST-IV bacteria cooperatively. T. vaginalis enhances paracellular permeability of hECs two times more than the individual bacterial species, by up to ∼10% versus ∼5%, respectively. However, any two or all three bacterial species are capable of synergizing this effect. T. vaginalis and the bacteria together increase the paracellular permeability of hECs by ∼50%, which is 5 to 10 times more than the results seen with the protozoan or bacteria alone. This effect is accompanied by enhancement of phosphatase activity, while phosphatase inhibition results in preservation of the integrity of the ectocervical cell monolayer. In addition, these microorganisms induce changes in the expression of tight junction proteins, particularly occludin, and of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α). Together, our findings establish that cooperative interactions between CST-IV bacteria and T. vaginalis enhance the paracellular permeability of the cervicovaginal epithelium by disturbing the integrity of the tight junction complex. Our study results highlight the importance of understanding the contribution of the vaginal microbiota to trichomoniasis.


Epithelial Cells/physiology , Microbial Interactions , Tight Junctions/physiology , Trichomonas vaginalis/physiology , Trichomonas vaginalis/pathogenicity , Vagina/physiology , Vaginosis, Bacterial/physiopathology , Female , Humans , Permeability
13.
Parasitology ; 146(9): 1156-1166, 2019 08.
Article En | MEDLINE | ID: mdl-30859930

Trichomonas vaginalis induces cellular damage to the host cells (cytotoxicity) through the proteolytic activity of multiple proteinases of the cysteine type (CPs). Some CPs are modulated by environmental factors such as iron, zinc, polyamines, etc. Thus, the goal of this study was to assess the effect of glucose on T. vaginalis cytotoxicity, proteolytic activity and the particular role of TvCP2 (TVAG_057000) during cellular damage. Cytotoxicity assays showed that glucose-restriction (GR) promotes the highest HeLa cell monolayers destruction (~95%) by trichomonads compared to those grown under high glucose (~44%) condition. Zymography and Western blot using different primary antibodies showed that GR increased the proteolytic activity, amount and secretion of certain CPs, including TvCP2. We further characterized the effect of glucose on TvCP2. TvCP2 increases in GR, localized in vesicles close to the plasma membrane and on the surface of T. vaginalis. Furthermore, pretreatment of GR-trichomonads with an anti-TvCP2r polyclonal antibody specifically reduced the levels of cytotoxicity and apoptosis induction to HeLa cells in a concentration-dependent manner. In conclusion, our data show that GR, as a nutritional stress condition, promotes trichomonal cytotoxicity to the host cells, increases trichomonad proteolytic activity and amount of CPs, such as TvCP2 involved in cellular damage.


Apoptosis , Cysteine Endopeptidases/metabolism , Glucose/metabolism , Trichomonas vaginalis/enzymology , Trichomonas vaginalis/pathogenicity , Cell Membrane/drug effects , Cell Membrane/metabolism , Glucose/pharmacology , HeLa Cells , Host-Parasite Interactions , Humans , Nutritional Physiological Phenomena , Proteolysis , Protozoan Proteins/metabolism
14.
Int J STD AIDS ; 30(5): 496-504, 2019 04.
Article En | MEDLINE | ID: mdl-30626281

Trichomonas vaginalis (TV) is a parasitic protozoan responsible for the sexually transmitted infection trichomoniasis. Trichomonas vaginalis virus (TVV) is a nonsegmented, 4.5-5 kbp, double-stranded RNA virus, from the Totiviridae family, which inhabits TV. A capsid protein consisting of 120 subunits is covered in channels aiding in RNA release. TVV is closely associated with the Golgi complex and is transmitted vertically. TVV has four subspecies, TVV1, TVV2, TVV3, and TVV4. The clinical significance of TVV and its effect on the pathogenicity of TV is not well known. We performed a systematic review of the literature on TVV to better understand its clinical significance and its role in the pathogenesis of TV.


Polymerase Chain Reaction/methods , RNA Viruses/classification , RNA Viruses/isolation & purification , RNA, Double-Stranded/genetics , RNA, Viral/genetics , Totiviridae/classification , Totiviridae/genetics , Trichomonas Infections/virology , Trichomonas vaginalis/virology , Capsid Proteins/genetics , Genome, Viral , Humans , Metronidazole , RNA Viruses/genetics , RNA Viruses/physiology , RNA, Double-Stranded/isolation & purification , RNA-Dependent RNA Polymerase/genetics , Totiviridae/isolation & purification , Trichomonas vaginalis/genetics , Trichomonas vaginalis/isolation & purification , Trichomonas vaginalis/pathogenicity
15.
PLoS Biol ; 17(1): e3000098, 2019 01.
Article En | MEDLINE | ID: mdl-30608924

Mitochondria originated from proteobacterial endosymbionts, and their transition to organelles was tightly linked to establishment of the protein import pathways. The initial import of most proteins is mediated by the translocase of the outer membrane (TOM). Although TOM is common to all forms of mitochondria, an unexpected diversity of subunits between eukaryotic lineages has been predicted. However, experimental knowledge is limited to a few organisms, and so far, it remains unsettled whether the triplet-pore or the twin-pore structure is the generic form of TOM complex. Here, we analysed the TOM complex in hydrogenosomes, a metabolically specialised anaerobic form of mitochondria found in the excavate Trichomonas vaginalis. We demonstrate that the highly divergent ß-barrel T. vaginalis TOM (TvTom)40-2 forms a translocation channel to conduct hydrogenosomal protein import. TvTom40-2 is present in high molecular weight complexes, and their analysis revealed the presence of four tail-anchored (TA) proteins. Two of them, Tom36 and Tom46, with heat shock protein (Hsp)20 and tetratricopeptide repeat (TPR) domains, can bind hydrogenosomal preproteins and most likely function as receptors. A third subunit, Tom22-like protein, has a short cis domain and a conserved Tom22 transmembrane segment but lacks a trans domain. The fourth protein, hydrogenosomal outer membrane protein 19 (Homp19) has no known homology. Furthermore, our data indicate that TvTOM is associated with sorting and assembly machinery (Sam)50 that is involved in ß-barrel assembly. Visualisation of TvTOM by electron microscopy revealed that it forms three pores and has an unconventional skull-like shape. Although TvTOM seems to lack Tom7, our phylogenetic profiling predicted Tom7 in free-living excavates. Collectively, our results suggest that the triplet-pore TOM complex, composed of three conserved subunits, was present in the last common eukaryotic ancestor (LECA), while receptors responsible for substrate binding evolved independently in different eukaryotic lineages.


Carrier Proteins/metabolism , Mitochondrial Membrane Transport Proteins/metabolism , Trichomonas vaginalis/metabolism , Carrier Proteins/genetics , Carrier Proteins/physiology , Membrane Proteins/metabolism , Membrane Transport Proteins/metabolism , Mitochondria/metabolism , Mitochondrial Precursor Protein Import Complex Proteins , Organelles , Phylogeny , Protein Transport/physiology , Trichomonas vaginalis/pathogenicity , Trichomonas vaginalis/physiology
16.
Parasitology ; 146(9): 1150-1155, 2019 08.
Article En | MEDLINE | ID: mdl-30616707

Trichomonas vaginalis is an anaerobic protist, responsible for the most prevalent non-viral sexually transmitted infection in humans. One of the most intriguing aspects of T. vaginalis pathobiology is the complex relationship with intracellular microbial symbionts: a group of dsRNA viruses belonging to family of Totiviridae (T. vaginalis virus), and eubacteria belonging to the Mycoplasma genus, in particular Mycoplasma hominis. Both microorganisms seem to strongly influence the lifestyle of T. vaginalis, suggesting a role of the symbiosis in the high variability of clinical presentation and sequelae during trichomoniasis. In the last few years many aspects of this unique symbiotic relationship have been investigated: M. hominis resides and replicates in the protozoan cell, and T. vaginalis is able to pass the bacterial infection to both mycoplasma-free protozoan isolates and human epithelial cells; M. hominis synergistically upregulates the proinflammatory response of human monocytes to T. vaginalis. Furthermore, the influence of M. hominis over T. vaginalis metabolism and physiology has been characterized. The identification of a novel species belonging to the class of Mollicutes (Candidatus Mycoplasma girerdii) exclusively associated to T. vaginalis opens new perspectives in the research of the complex series of events taking place in the multifaceted world of the vaginal microbiota, both under normal and pathological conditions.


Mycoplasma Infections/microbiology , Mycoplasma hominis/physiology , Symbiosis , Trichomonas Vaginitis/parasitology , Trichomonas vaginalis/pathogenicity , Female , Humans , Inflammation , Microbiota , Mycoplasma hominis/immunology , Sexually Transmitted Diseases/immunology , Sexually Transmitted Diseases/parasitology , Totiviridae/metabolism , Trichomonas vaginalis/immunology , Vagina/microbiology , Vagina/parasitology
17.
Int J Cancer ; 144(10): 2377-2380, 2019 05 15.
Article En | MEDLINE | ID: mdl-30242839

We previously observed a positive association between seropositivity for the parasite Trichomonas vaginalis and risk of clinically significant prostate cancer at diagnosis. Here, we examined whether T. vaginalis seropositivity was associated with increased prostate cancer-specific or all-cause mortality among prostate cancer patients. We studied 736 men with prostate cancer from the Physicians' Health Study (PHS) and 749 men with prostate cancer from the Health Professionals Follow-Up Study (HPFS). We used Cox proportional hazards regression models to estimate multivariable-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) of the association between T. vaginalis serostatus and progression to death from prostate cancer and from all causes. In PHS, 423 men died of any cause during a median follow-up of 13.8 years from the date of cancer diagnosis, among whom 131 died of prostate cancer. In HPFS, there were 287 deaths, including 77 deaths from prostate cancer, during a median follow-up of 12.8 years. We found no association between T. vaginalis serostatus and either prostate cancer mortality or all-cause mortality in either the PHS or HPFS. While previous studies suggest a possible role for T. vaginalis in the development of clinically significant prostate cancer, our findings do not support the hypothesis that T. vaginalis serostatus is associated with mortality among prostate cancer patients.


Prostatic Neoplasms/etiology , Prostatic Neoplasms/mortality , Trichomonas Vaginitis/complications , Trichomonas vaginalis/pathogenicity , Aged , Case-Control Studies , Disease Progression , Female , Follow-Up Studies , Humans , Male , Middle Aged , Prospective Studies , Prostate/parasitology , Prostate/pathology , Prostatic Neoplasms/parasitology , Prostatic Neoplasms/pathology , Risk Factors , Trichomonas Vaginitis/pathology
18.
Clin Infect Dis ; 68(9): 1575-1580, 2019 04 24.
Article En | MEDLINE | ID: mdl-30395193

BACKGROUND: Because of its increasing prevalence worldwide, its sexual transmissibility and its facilitation of human immunodeficiency virus transmission, Trichomonas vaginalis (TV) infection constitutes an important public health concern. THE AIM OF THE STUDY: While searching for possible resistant TV cases, adequacy of management of TV-infected women was assessed. METHODS: Cervical cytology between July 2007 and July 2014 was tested with TV polymerase chain reaction, and 304 women expressed repeatedly positive results, 718 in total. For each of these positive results, a questionnaire about treatment decisions was sent to the 182 Belgian physicians treating these women. RESULTS: From the 346 returned questionnaires by their physician it was evident that 58.1% of women with repeatedly positive TV had received no treatment. TV was overlooked in 31.5%, and in 17.6% the test result was seen but ignored. Upon seeing the positive result, 23.9% of physicians decided that this finding was not important enough to institute treatment, and/or requested confirmatory tests. Adequate treatment was prescribed in 38.4%. Retreatment after failed therapy was given in only 29.3% of the cases. And 60% of the partners of women with persistent TV infection were not traced, nor treated. CONCLUSION: Most of the repeatedly positive TV infection may not be due to antibiotics resistance. The low awareness, poor attention, failure of contact tracing, and low rates of proper treatment provided by treating physicians question the adequacy of the current management of TV infection and requires renewed education campaigns and increased surveillance.


Attitude of Health Personnel , Health Knowledge, Attitudes, Practice , Trichomonas Vaginitis/drug therapy , Trichomonas Vaginitis/psychology , Trichomonas vaginalis/pathogenicity , Adult , Antiprotozoal Agents/therapeutic use , Belgium , Female , Humans , Metronidazole/therapeutic use , Middle Aged , Practice Patterns, Physicians'/statistics & numerical data , Retrospective Studies , Sexual Partners/psychology , Surveys and Questionnaires , Tinidazole/therapeutic use , Treatment Outcome , Trichomonas Vaginitis/parasitology , Trichomonas vaginalis/drug effects , Trichomonas vaginalis/growth & development
19.
Georgian Med News ; (297): 41-46, 2019 Dec.
Article Ru | MEDLINE | ID: mdl-32011293

Objective - study of the features of Trichomonas vaginalis invasion in pregnant women and newborns. The cultures of Trichomonas isolated from a pregnant woman, her newborn girl, as well as amniotic fluid were examined. The ultrastructure of the cells was studied using a TEM-125K microscope equipped with a SAI-01A system (SELMI), using a DX 2 CCD camera and the KAPPA software package. The verification of STI pathogens was carried out by PCR, in particular, Trichomonas tenax and Pentatrichomonas hominis were identified by experimental original primers that were developed using real-time PCR (PCR-RT). The invasion of Trichomonas vaginalis of the genital tract of a newborn girl, amniotic fluid and amniotic membrane is characterized clinically and instrumentally. We proved the possibility of Trichomonas vaginalis invasion of newborn children not only during the passage of the child through the infected birth canal, but also due to a defect in the fetal membranes with the development of chorioamnionitis, followed by infection of the amniotic fluid and possible infection of the fetus.


Infectious Disease Transmission, Vertical , Pregnancy Complications, Infectious , Trichomonas Vaginitis , Trichomonas vaginalis , Trichomonas , Female , Humans , Infant, Newborn , Polymerase Chain Reaction , Pregnancy , Trichomonas Vaginitis/diagnosis , Trichomonas Vaginitis/transmission , Trichomonas vaginalis/isolation & purification , Trichomonas vaginalis/pathogenicity , Vagina/parasitology
20.
Parasitol Res ; 117(9): 2735-2742, 2018 Sep.
Article En | MEDLINE | ID: mdl-29936622

A close association between Trichomonas vaginalis (TV) infection and bacterial vaginosis (BV) has been reported. Some other studies have found association is stronger with intermediate Nugent score than BV. Most studies have used wet mount microscopy, a relatively insensitive method, to detect TV infection. We wanted to study the association of TV infection with BV and with intermediate Nugent score. We undertook a cross-sectional hospital-based study of 1110 non-pregnant women from Odisha state, India, aged between 18 and 45 years, collecting vaginal swabs for diagnosis of BV by Nugent score (NS) criteria and TV by PCR analysis. TV infection was found in 13.3% of women with intermediate Nugent score (NS 4-6) and 13.6% with BV (NS 7-10). Before adjustment, TV infection was associated with BV, intermediate Nugent, vaginal pH ≥ 4.5, and age group between 26 and 35 years. Multivariate analysis confirmed that TV infection was more likely to have raised vaginal pH, either BV or intermediate Nugent. Proportion of TV cases increased sequentially with the increase in Nugent score up to NS 6, after which a decline was observed. Vaginal pH was higher in the TV-infected group than the uninfected group in women with intermediate Nugent, but no difference was noticed in women with BV. TV infection was equally prevalent in women with intermediate Nugent as well as BV. In the intermediate Nugent group women, TV infection was found only when vaginal pH was raised, indicating a crucial role of vaginal pH in determining TV infection.


Trichomonas Vaginitis/diagnosis , Trichomonas Vaginitis/pathology , Trichomonas vaginalis/pathogenicity , Vagina/physiology , Adolescent , Adult , Cross-Sectional Studies , Female , Humans , Hydrogen-Ion Concentration , India , Middle Aged , Prevalence , Trichomonas Vaginitis/parasitology , Vagina/parasitology , Young Adult
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