RESUMEN
Trichomonas vaginalis is an amitochondriate protozoan and the agent of human trichomoniasis, the most prevalent non-viral sexually transmitted infection (STI) in the world. In this study we showed that 2,4-diamine-quinazoline derivative compound (PH100) kills T. vaginalis. PH100 showed activity against fresh clinical and American Type Culture Collection (ATCC) T. vaginalis isolates with no cytotoxicity against cells (HMVI, 3T3-C1 and VERO) and erythrocytes. In addition, PH100 showed synergistic action with metronidazole, indicating that these compounds act by different mechanisms. When investigating the mechanism of action of PH100 to ATCC 30236, apoptosis-like characteristics were observed, such as phosphatidylserine exposure, membrane alterations, and modulation of gene expression and activity of peptidases related to apoptosis. The apoptosis-like cell death features were not observed for the fresh clinical isolate treated with PH100 revealing distinct profiles. Our data revealed the heterogeneity among T. vaginalis isolates and contribute with the understanding of mechanisms of cell death in pathogenic eukaryotic organisms without mitochondria.
Asunto(s)
Diaminas/farmacología , Parásitos/efectos de los fármacos , Péptido Hidrolasas/metabolismo , Quinazolinas/farmacología , Vaginitis por Trichomonas/tratamiento farmacológico , Trichomonas vaginalis/efectos de los fármacos , Células 3T3 , Animales , Apoptosis/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Línea Celular , Chlorocebus aethiops , Femenino , Humanos , Metronidazol/farmacología , Ratones , Vaginitis por Trichomonas/parasitología , Células VeroRESUMEN
Trichomonas vaginalis is responsible for the most common non-viral, sexually transmitted infection, human trichomoniasis, and is associated with an increased susceptibility to HIV. An escalation in resistance (2.5-10%) to the clinical drug, metronidazole (MTZ), has been detected and this compound also has adverse side-effects. Therefore, new treatment options are urgently required. Herein, we investigate the possible anti-T. vaginalis activity of 1,10-phenanthroline-5,6-dione (phendione) and its metal complexes, [Ag(phendione)2]ClO4 and [Cu(phendione)3](ClO4)2·4H2O. Minimum inhibitory concentration (MIC) against T. vaginalis ATCC 30236 and three fresh clinical isolates and mammalian cells were performed using serial dilution generating IC50 and CC50 values. Drugs combinations with MTZ were evaluated by chequerboard assay. A strong anti-T. vaginalis activity was found for all test compounds. IC50 values obtained for [Cu(phendione)3](ClO4)2·4H2O were similar or lower than those obtained for MTZ. In vitro assays with normal cells showed low cytotoxicity and [Cu(phendione)3](ClO4)2·4H2O presented a high selectivity index (SI) for fibroblasts (SI = 11.39) and erythrocytes (SI > 57.47). Chequerboard assay demonstrated that the combination of [Cu(phendione)3](ClO4)2·4H2O with MTZ leads to synergistic interaction, which suggests distinct mechanisms of action of the copper-phendione complex and avoiding the MTZ resistance pathways. Our results highlight the importance of phendione-based drugs as potential molecules of pharmaceutical interest.
Asunto(s)
Metronidazol/farmacología , Fenantrolinas/química , Fenantrolinas/farmacología , Trichomonas vaginalis/efectos de los fármacos , Animales , Línea Celular , Sinergismo Farmacológico , Eritrocitos/efectos de los fármacos , Femenino , Fibroblastos/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Ratones , Pruebas de Sensibilidad Microbiana , Vaginitis por Trichomonas/parasitologíaRESUMEN
INTRODUCTION: The challenges related to the diagnosis of sexually transmitted infections present more complex factors in remote and hard-to-reach areas. The use of self-collection devices that facilitate the obtaining of a biological sample with high quality for sensitive molecular tests have been examined. This study aimed to evaluate the performance and acceptance of the Evalyn® Brush (Rovers® Medical Devices) for detection of T. vaginalis among women living in the riverside communities of Amazonas, Brazil. METHODOLOGY: The study included 300 riverside women. They received instructions for self-collection, carried out the task, and then answered a questionnaire on the use of the device. T. vaginalis was detected by Polymerase Chain Reaction, using primers TVK3/TVK7. RESULTS: The mean age of the women was 35.8 years, and most of them presented low schooling, low income, agricultural activity and lived in a marital union. All samples were positive for human genomic DNA (100%) and the prevalence of T. vaginalis infection was 5.6% (n = 17). Of the 300 women, 293 (97.7%) indicated that they liked the use of the device, 287 (95.7%) reported having had no difficulty in handling it, 265 (88.3%) did not feel any type of discomfort and 228 (76%) said they preferred the self-collection to the collection made by the professional, mainly due to privacy and comfort. CONCLUSIONS: The Evalyn® Brush proved reliable as a device for the collection of biological samples for molecular analysis and was well-accepted by women. Its use can be indicated in remote and hard to reach places.
Asunto(s)
Manejo de Especímenes/instrumentación , Vaginitis por Trichomonas/parasitología , Adulto , Anciano , Brasil/epidemiología , Estudios Transversales , Diseño de Equipo , Femenino , Humanos , Persona de Mediana Edad , Prevalencia , Autocuidado , Parejas Sexuales , Manejo de Especímenes/métodos , Vaginitis por Trichomonas/epidemiología , Trichomonas vaginalis/genética , Trichomonas vaginalis/aislamiento & purificación , Adulto JovenRESUMEN
Trichomonas vaginalis is an extracellular parasite that binds to the epithelium of the human urogenital tract and causes the sexually transmitted infection, trichomoniasis. In view of increased resistance to drugs belonging to the 5-nitroimidazole class, new treatment alternatives are urgently needed. In this study, eight semisynthetized triterpene derivatives were evaluated for in vitro anti-T. vaginalis activity. Ursolic acid and its derivative, 3-oxime-urs-12-en-28-oic-ursolic acid (9), presented the best anti-T. vaginalis activity when compared to other derivatives, with minimum inhibitory concentration (MIC) at 25 µM. Moreover, 9 was active against several T. vaginalis fresh clinical isolates. Hemolysis assay demonstrated that 9 presented a low hemolytic effect. Importantly, 25 µM 9 was not cytotoxic against the Vero cell lineage. Finally, we demonstrated that compound 9 acts synergistically with metronidazole against a T. vaginalis metronidazole-resistant isolate. This report reveals the high potential of the triterpenoid derivative 9 as trichomonicidal agent.
Asunto(s)
Antitricomonas/farmacología , Sinergismo Farmacológico , Metronidazol/farmacología , Tricomoniasis/tratamiento farmacológico , Vaginitis por Trichomonas/tratamiento farmacológico , Trichomonas vaginalis/efectos de los fármacos , Triterpenos/farmacología , Animales , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Chlorocebus aethiops , Resistencia a Medicamentos , Quimioterapia Combinada , Femenino , Células HeLa , Hemólisis/efectos de los fármacos , Humanos , Pruebas de Sensibilidad Microbiana , Tricomoniasis/parasitología , Vaginitis por Trichomonas/parasitología , Triterpenos/química , Células Vero , Ácido UrsólicoRESUMEN
OBJECTIVE: The aim of this study was to assess the correlation between atypical squamous cells (ASC) and inflammatory infiltrate and vaginal microbiota using cervical liquid-based cytological (SurePath®) and high-risk human papillomavirus (HR-HPV) tests. STUDY DESIGN: A cross-sectional study was conducted using a 6-year database from a laboratory in Fortaleza (Brazil). Files from 1,346 ASC cases were divided into subgroups and results concerning inflammation and vaginal microorganisms diagnosed by cytology were compared with HR-HPV test results. RESULTS: An absence of specific microorganisms (ASM) was the most frequent finding (ASC of undetermined significance, ASC-US = 74%; ASC - cannot exclude high-grade squamous intraepithelial lesion, ASC-H = 68%), followed by bacterial vaginosis (ASC-US = 20%; ASC- H = 25%) and Candida spp. (ASC-US = 6%; ASC-H = 5%). Leukocyte infiltrate was present in 71% of ASC-US and 85% of ASC-H (p = 0.0040), and in these specific cases HR-HPV tests were positive for 65 and 64%, respectively. A positive HR-HPV test was relatively more frequent when a specific microorganism was present, and Candida spp. was associated with HR-HPV-positive results (p = 0.0156), while an ASM was associated with negative HR-HPV results (p = 0.0370). CONCLUSION: ASC-US is associated with an absence of inflammation or vaginosis, while ASC-H smears are associated with Trichomonas vaginalis and inflammatory infiltrate. A positive HR-HPV is associated with Candida spp. in ASC cytology.
Asunto(s)
Células Escamosas Atípicas del Cuello del Útero/patología , Pruebas de ADN del Papillomavirus Humano , Técnicas Microbiológicas , Papillomaviridae/genética , Infecciones por Papillomavirus/virología , ARN Viral/genética , Lesiones Intraepiteliales Escamosas de Cuello Uterino/patología , Neoplasias del Cuello Uterino/patología , Vaginitis/patología , Adulto , Células Escamosas Atípicas del Cuello del Útero/microbiología , Células Escamosas Atípicas del Cuello del Útero/parasitología , Células Escamosas Atípicas del Cuello del Útero/virología , Brasil , Candida/aislamiento & purificación , Candidiasis Vulvovaginal/microbiología , Candidiasis Vulvovaginal/patología , Estudios Transversales , Bases de Datos Factuales , Femenino , Humanos , Leucocitos/patología , Biopsia Líquida , Persona de Mediana Edad , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/patología , Valor Predictivo de las Pruebas , ARN Viral/aislamiento & purificación , Factores de Riesgo , Lesiones Intraepiteliales Escamosas de Cuello Uterino/microbiología , Lesiones Intraepiteliales Escamosas de Cuello Uterino/parasitología , Lesiones Intraepiteliales Escamosas de Cuello Uterino/virología , Vaginitis por Trichomonas/parasitología , Vaginitis por Trichomonas/patología , Trichomonas vaginalis/aislamiento & purificación , Neoplasias del Cuello Uterino/microbiología , Neoplasias del Cuello Uterino/parasitología , Neoplasias del Cuello Uterino/virología , Frotis Vaginal , Vaginitis/microbiología , Vaginitis/parasitología , Adulto JovenRESUMEN
Trichomonas vaginalis is a common sexually transmitted parasite that colonizes the human urogenital tract, where it remains extracellular and adheres to epithelial cells. Infections range from asymptomatic to highly inflammatory, depending on the host and the parasite strain. Despite the serious consequences associated with trichomoniasis disease, little is known about parasite or host factors involved in attachment of the parasite-to-host epithelial cells. Here, we report the identification of microvesicle-like structures (MVs) released by T. vaginalis. MVs are considered universal transport vehicles for intercellular communication as they can incorporate peptides, proteins, lipids, miRNA, and mRNA, all of which can be transferred to target cells through receptor-ligand interactions, fusion with the cell membrane, and delivery of a functional cargo to the cytoplasm of the target cell. In the present study, we demonstrated that T. vaginalis release MVs from the plasma and the flagellar membranes of the parasite. We performed proteomic profiling of these structures demonstrating that they possess physical characteristics similar to mammalian extracellular vesicles and might be selectively charged with specific protein content. In addition, we demonstrated that viable T. vaginalis parasites release large vesicles (LVs), membrane structures larger than 1 µm that are able to interact with other parasites and with the host cell. Finally, we show that both populations of vesicles present on the surface of T vaginalis are induced in the presence of host cells, consistent with a role in modulating cell interactions.
Asunto(s)
Vesículas Extracelulares/metabolismo , Interacciones Huésped-Parásitos , Vaginitis por Trichomonas/metabolismo , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/fisiología , Trichomonas vaginalis/ultraestructura , Comunicación Celular , Vesículas Extracelulares/química , Vesículas Extracelulares/ultraestructura , Femenino , Células HeLa , Humanos , Proteómica , Proteínas Protozoarias/análisis , Proteínas Protozoarias/metabolismo , Trichomonas vaginalis/química , Trichomonas vaginalis/citologíaRESUMEN
Trichomonas vaginalis is a protozoan parasite that can adapt to the trichomonicidal Zn2+ concentrations of the male urogenital tract microenvironment. This adaptation is mediated by molecular mechanisms, including proteinase expression, that are regulated by cations such as Zn2+. Herein, we characterized the previously identified 50kDa metalloproteinase aminopeptidase P (M24 family) member TvMP50 as a new Zn2+-mediated parasite virulence factor. Quantitative RT-PCR and indirect immunofluorescence assays corroborated the positive regulation of both mp50 gene expression and native TvMP50 protein overexpression in the cytoplasm and secretion products of parasites grown in the presence of Zn2+. Furthermore, this active metalloproteinase was characterized as a new virulence factor by assaying cytotoxicity toward prostatic DU145 cell monolayers as well as the inhibition of parasite and secreted soluble protein proteolytic activity in the 50kDa proteolytic region by the specific metalloproteinase inhibitor 1,10-phenanthroline and the chelating agents EDTA and EGTA. Parasite and secreted soluble protein cytotoxicity toward DU145 cells were reduced by treatment with an α-rTvMP50 polyclonal antibody. Our results show that the metalloproteinase TvMP50 is a new virulence factor modulated by Zn2+, which is present during male trichomoniasis, possibly explaining T. vaginalis survival even within the adverse conditions of the male urogenital microenvironment.
Asunto(s)
Metaloproteasas/metabolismo , Proteínas Protozoarias/metabolismo , Trichomonas vaginalis/enzimología , Factores de Virulencia/metabolismo , Zinc/metabolismo , Línea Celular , Células Cultivadas , Cromatografía Liquida , Femenino , Expresión Génica , Humanos , Masculino , Metaloproteasas/química , Metaloproteasas/genética , Transporte de Proteínas , Proteínas Protozoarias/química , Proteínas Protozoarias/genética , Espectrometría de Masas en Tándem , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/genética , Trichomonas vaginalis/patogenicidad , Factores de Virulencia/química , Factores de Virulencia/genéticaRESUMEN
Trichomonas vaginalis causes trichomoniasis, a neglected sexually transmitted disease. Due to severe health consequences and treatment failure, new therapeutic alternatives are crucial. Phloroglucinols from southern Brazilian Hypericum species demonstrated anti-T. vaginalis and anti-Leishmania amazonensis activities. The modulation of biochemical pathways involved in the control of inflammatory response by ectonucleotidases, NTPDase, and ecto-5'-nucleotidase represents new targets for combating protozoa. This study investigated the activity of phloroglucinol derivatives of Hypericum species from southern Brazil against T. vaginalis as well as its ability on modulating parasite ectonucleotidases and, consequently, immune parameters through ATP and adenosine effects. Phloroglucinol derivatives screening revealed activity for isoaustrobrasilol B (IC50 38 µm) with no hemolytic activity. Although the most active compound induced cytotoxicity against a mammalian cell lineage, the in vivo model evidenced absence of toxicity. Isoaustrobrasilol B significantly inhibited NTPDase and ecto-5'-nucleotidase activities, and the immune modulation attributed to extracellular nucleotide accumulation was evaluated. The production of ROS and IL-6 by T. vaginalis-stimulated neutrophils was not affected by the treatment. Conversely, IL-8 levels were significantly enhanced. The associative mechanism of trophozoites death and ectonucleotidases modulation by isoaustrobrasilol B may increase the susceptibility of T. vaginalis to host innate immune cell like neutrophils consequently, contributing to parasite clearance.
Asunto(s)
Antiparasitarios/farmacología , Floroglucinol/análogos & derivados , Vaginitis por Trichomonas/tratamiento farmacológico , Trichomonas vaginalis/efectos de los fármacos , Antiparasitarios/química , Línea Celular , Femenino , Humanos , Hidrólisis/efectos de los fármacos , Hypericum/química , Nucleótidos/metabolismo , Floroglucinol/química , Floroglucinol/farmacología , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/fisiologíaRESUMEN
Sexually transmitted infections are an important cause of morbidity among sexually active women worldwide, and have been implicated as cofactors in the pathogenesis of cervical cancer. We investigated the prevalence of human papillomavirus (HPV), Chlamydia trachomatis (CT), and Trichomonas vaginalis (TV), and accessed the diversity of HPV in women with normal and abnormal cytology in Manaus, Brazil. We used polymerase chain reaction and HPV genotyping by direct sequencing. The chi-square test was used to calculate the absolute and relative frequencies of the categorical variables, and Fisher's test was used when P < 0.05. The level of significance was set at 5%. All statistical analyses were performed using R 2.9.0. There were statistically significant differences in age (P = 0.0395), education level (P = 0.0131), sexual partners (P = 0.0211), condom use (P = 0.0039), marital status (P < 0.0001), and pregnancy (P = 0.0003) between the normal and abnormal groups. HPV DNA was found in 36.56 and 93.88% of subjects in the normal and abnormal groups, respectively. A total of 19 genotypes were detected; HPV16 was the most common, followed by HPV58. The percentages of TV and CT DNA were 18.04 and 9.02% in the normal group, respectively. The percentages of HPV/TV and HPV/CT coinfection were 12.5% each in women with normal cytology. These findings improve our understanding of HPV, CT, and TV, and the distribution of HPV types, which may be relevant to vaccination strategies for protecting women from the north of Brazil from cervical cancers and precancerous lesions.
Asunto(s)
Infecciones por Chlamydia/epidemiología , Chlamydia trachomatis/genética , Genotipo , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , Vaginitis por Trichomonas/epidemiología , Trichomonas vaginalis/genética , Adolescente , Adulto , Brasil , Infecciones por Chlamydia/microbiología , Infecciones por Chlamydia/patología , Chlamydia trachomatis/aislamiento & purificación , Femenino , Humanos , Persona de Mediana Edad , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/patología , Infecciones por Papillomavirus/virología , Prevalencia , Vaginitis por Trichomonas/parasitología , Vaginitis por Trichomonas/patología , Trichomonas vaginalis/aislamiento & purificaciónRESUMEN
Trichomonas vaginalis is a common sexually transmitted parasite that colonizes the human urogenital tract. Infections range from asymptomatic to highly inflammatory, depending on the host and the parasite strain. Different T. vaginalis strains vary greatly in their adherence and cytolytic capacities. These phenotypic differences might be attributed to differentially expressed genes as a consequence of extra-genetic variation, such as epigenetic modifications. In this study, we explored the role of histone acetylation in regulating gene transcription and pathogenesis in T. vaginalis. Here, we show that histone 3 lysine acetylation (H3KAc) is enriched in nucleosomes positioned around the transcription start site of active genes (BAP1 and BAP2) in a highly adherent parasite strain; compared with the low acetylation abundance in contrast to that observed in a less-adherent strain that expresses these genes at low levels. Additionally, exposition of less-adherent strain with a specific histone deacetylases inhibitor, trichostatin A, upregulated the transcription of BAP1 and BAP2 genes in concomitance with an increase in H3KAc abundance and chromatin accessibility around their transcription start sites. Moreover, we demonstrated that the binding of initiator binding protein, the transcription factor responsible for the initiation of transcription of ~75% of known T. vaginalis genes, depends on the histone acetylation state around the metazoan-like initiator to which initiator binding protein binds. Finally, we found that trichostatin A treatment increased parasite aggregation and adherence to host cells. Our data demonstrated for the first time that H3KAc is a permissive histone modification that functions to mediate both transcription and pathogenesis of the parasite T. vaginalis.
Asunto(s)
Adhesión Celular/efectos de los fármacos , Agregación Celular/efectos de los fármacos , Histonas/metabolismo , Vaginitis por Trichomonas/patología , Trichomonas vaginalis/genética , Trichomonas vaginalis/patogenicidad , Acetilación/efectos de los fármacos , Adhesión Celular/genética , Adhesión Celular/fisiología , Agregación Celular/fisiología , Línea Celular Tumoral , Cuello del Útero/citología , Cuello del Útero/metabolismo , Cuello del Útero/parasitología , Cromatina/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Femenino , Regulación de la Expresión Génica , Células HeLa , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Ácidos Hidroxámicos/farmacología , Metaloendopeptidasas/genética , Unión Proteica/fisiología , Proteínas Protozoarias/genética , Proteínas Protozoarias/metabolismo , Transcripción Genética/genética , Activación Transcripcional/genética , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/metabolismoRESUMEN
The aim of this study was to evaluate different methods for Trichomonas vaginalis diagnosis during pregnancy in order to prevent maternal and perinatal complications. A total of 386 vaginal exudates from pregnant women were analyzed. T. vaginalis was investigated by 3 types of microscopic examinations direct wet mount with physiologic saline solution, prolonged May-Grunwald Giemsa (MGG) staining, and wet mount with sodium-acetate-formalin (SAF)/methylene blue method. PCR for 18S rRNA gene as well as culture in liquid medium were performed. The sensitivity and specificity of the microscopic examinations were evaluated considering the culture media positivity or the PCR techniques as gold standard. The frequency of T. vaginalis infection was 6.2% by culture and/or PCR, 5.2% by PCR, 4.7% by culture, 3.1% by SAF/methylene blue method and 2.8% by direct wet smear and prolonged MGG staining. The sensitivities were 83.3%, 75.0%, 50.0%, and 45.8% for PCR, culture, SAF/methylene blue method, and direct wet smear-prolonged MGG staining, respectively. The specificity was 100% for all the assessed methods. Microscopic examinations showed low sensitivity, mainly in asymptomatic pregnant patients. It is necessary to improve the detection of T. vaginalis using combined methods providing higher sensitivity, such as culture and PCR, mainly in asymptomatic pregnant patients, in order to prevent maternal and perinatal complications.
Asunto(s)
Reacción en Cadena de la Polimerasa/métodos , Coloración y Etiquetado/métodos , Vaginitis por Trichomonas/diagnóstico , Trichomonas vaginalis/aislamiento & purificación , Argentina , Eosina Amarillenta-(YS) , Femenino , Hospitales Universitarios , Humanos , Azul de Metileno , Microscopía/métodos , Embarazo , Complicaciones Parasitarias del Embarazo/prevención & control , Sensibilidad y Especificidad , Vaginitis por Trichomonas/parasitología , Frotis Vaginal/métodosRESUMEN
BACKGROUND: Trichomonas vaginalis infection (TVI) is one of the most common sexually transmitted diseases in the United States. We sought to determine the features of TVI in a referral-based vaginitis center, focusing on diagnosis and treatment of difficult cases. METHODS: We conducted a retrospective review of all patients with TVI, based on International Classification of Diseases, Ninth Revision codes, seen at the Drexel Vaginitis Center between January 2008 and November 2013. Information collected on each subject included demographics, symptoms, examination findings, diagnostic tests, and treatment regimens. RESULTS: Of approximately 4000 new patient visits during our study period, 80 subjects were identified with TVI. Twenty subjects presented with known TVI, with most having clinically resistant infections. Diagnosis was confirmed by saline microscopy in 45%, OSOM rapid test in 40%, and clinical history in the remaining 15%. Treatment regimens varied: 20% received single 2-g dosing of either metronidazole or tinidazole, 50% received high-dose regimens, 20% received therapy with vaginal paromomycin, and 10% underwent desensitization for nitroimidazole allergy. Sixty subjects had newly diagnosed TVI, with 35% diagnosed by saline microscopy, 41.7% by OSOM rapid test, and 23.3% by APTIMA. Treatment regimens for these subjects included single 2-g dosing in 88.3%, high-dose regimen in 8.3%, and other formulations in the remaining 3.4%. In total, 80% of our subjects returned for follow-up; all of whom were cured. CONCLUSIONS: T. vaginalis infection is a rare condition in a tertiary care vaginitis center and often requires nonstandard treatments. Among those who returned for follow-up, the cure rate was 100%.
Asunto(s)
Enfermedades de Transmisión Sexual/diagnóstico , Vaginitis por Trichomonas/diagnóstico , Trichomonas vaginalis/aislamiento & purificación , Vagina/parasitología , Adolescente , Adulto , Anciano , Alquilantes , Antiinfecciosos/uso terapéutico , Femenino , Humanos , Metronidazol/uso terapéutico , Microscopía , Persona de Mediana Edad , Estudios Retrospectivos , Enfermedades de Transmisión Sexual/tratamiento farmacológico , Enfermedades de Transmisión Sexual/parasitología , Centros de Atención Terciaria , Tinidazol/uso terapéutico , Vaginitis por Trichomonas/tratamiento farmacológico , Vaginitis por Trichomonas/parasitología , Adulto JovenRESUMEN
Trichomonas vaginalis is the etiological agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in world, with 276.4 million new cases each year. T. vaginalis can be naturally infected with Mycoplasma hominis and Trichomonasvirus species. This study aimed to evaluate the prevalence of T. vaginalis infected with four distinct T. vaginalis viruses (TVVs) and M. hominis among isolates from patients in Porto Alegre city, South Brazil. An additional goal of this study was to investigate whether there is association between metronidazole resistance and the presence of M. hominis during TVV infection. The RNA expression level of the pyruvate ferredoxin oxidoreductase (PFOR) gene was also evaluated among metronidazole-resistant and metronidazole-sensitive T. vaginalis isolates. A total of 530 urine samples were evaluated, and 5.7% samples were positive for T. vaginalis infection. Among them, 4.51% were isolated from female patients and 1.12% were from male patients. Remarkably, the prevalence rates of M. hominis and TVV-positive T. vaginalis isolates were 56.7% and 90%, respectively. Most of the T. vaginalis isolates were metronidazole-sensitive (86.7%), and only four isolates (13.3%) were resistant. There is no statistically significant association between infection by M. hominis and infection by TVVs. Our results refute the hypothesis that the presence of the M. hominis and TVVs is enough to confer metronidazole resistance to T. vaginalis isolates. Additionally, the role of PFOR RNA expression levels in metronidazole resistance as the main mechanism of resistance to metronidazole could not be established. This study is the first report of the T. vaginalis infection by M. hominis and TVVs in a large collection of isolates from South Brazil.
Asunto(s)
Mycoplasma hominis/aislamiento & purificación , Virus ARN/aislamiento & purificación , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/virología , Adolescente , Adulto , Anciano , Antiprotozoarios/farmacología , Secuencia de Bases , Brasil , Resistencia a Medicamentos , Femenino , Humanos , Masculino , Metronidazol/farmacología , Persona de Mediana Edad , Datos de Secuencia Molecular , Tipificación Molecular , Mycoplasma hominis/genética , Virus ARN/genética , Análisis de Secuencia de ADN , Vaginitis por Trichomonas/orina , Trichomonas vaginalis/efectos de los fármacos , Trichomonas vaginalis/microbiología , Adulto JovenRESUMEN
Human trichomonosis, infection with Trichomonas vaginalis, is the most common non-viral sexually transmitted disease in the world. The host-parasite interaction and pathophysiological processes of trichomonosis remain incompletely understood. This review focuses on the advancements reached in the area of the pathogenesis of T. vaginalis, especially in the role of the cysteine proteinases. It highlights various approaches made in this field and lists a group of trichomonad cysteine proteinases involved in diverse processes such as invasion of the mucous layer, cytoadherence, cytotoxicity, cytoskeleton disruption of red blood cells, hemolysis, and evasion of the host immune response. A better understanding of the biological roles of cysteine proteinases in the pathogenesis of this parasite could be used in the identification of new chemotherapeutic targets. An additional advantage could be the development of a vaccine in order to reduce transmission of T. vaginalis.
Asunto(s)
Proteasas de Cisteína/fisiología , Proteínas Protozoarias/fisiología , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/enzimología , Vacunas Bacterianas , Líquidos Corporales/parasitología , Adhesión Celular , Membrana Celular/parasitología , Femenino , Hemólisis , Interacciones Huésped-Parásitos , Humanos , Evasión Inmune , Hierro/metabolismo , Lisosomas/parasitología , Terapia Molecular Dirigida , Membrana Mucosa/parasitología , Membrana Mucosa/patología , Vaginitis por Trichomonas/patología , Trichomonas vaginalis/inmunología , Trichomonas vaginalis/patogenicidad , Vagina/parasitología , Vagina/patología , VirulenciaRESUMEN
Trichomonas vaginalis is a sexually transmitted flagellated protist parasite responsible for trichomoniasis. This parasite is dependent on high levels of iron, favoring its growth and multiplication. Iron also differentially regulates some trichomonad virulence properties by unknown mechanisms. However, there is evidence to support the existence of gene regulatory mechanisms at the transcriptional and posttranscriptional levels that are mediated by iron concentration in T. vaginalis. Thus, the goal of this study was to identify an RNA-binding protein in T. vaginalis that interacts with the tvcp4 RNA stem-loop structure, which may participate in a posttranscriptional iron regulatory mechanism mediated by RNA-protein interactions. We performed RNA electrophoretic mobility shift assay (REMSA) and supershift, UV cross-linking, Northwestern blot, and western blot (WB) assays using cytoplasmic protein extracts from T. vaginalis with the tvcp4 RNA hairpin structure as a probe. We identified a 135-kDa protein isolated by the UV cross-linking assays as α-actinin 3 (TvACTN3) by MALDI-TOF-MS that was confirmed by LS-MS/MS and de novo sequencing. TvACTN3 is a cytoplasmic protein that specifically binds to hairpin RNA structures from trichomonads and humans when the parasites are grown under iron-depleted conditions. Thus, TvACTN3 could participate in the regulation of gene expression by iron in T. vaginalis through a parallel posttranscriptional mechanism similar to that of the IRE/IRP system.
Asunto(s)
Actinina/metabolismo , Hierro/metabolismo , Vaginitis por Trichomonas/metabolismo , Trichomonas vaginalis/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Hierro/química , ARN/química , Procesamiento Postranscripcional del ARN/genética , Proteínas de Unión al ARN/metabolismo , Vaginitis por Trichomonas/parasitología , Vaginitis por Trichomonas/patologíaRESUMEN
This study describes the prevalence of infection by Chlamydia trachomatis, Neisseria gonorrhoeae and Trichomonas vaginalis in a female population in Amazonas, Brazil. We collected cervical samples from 361 women examined at 10 primary care health services in the city of Coari, Amazonas, Brazil. The women were interviewed about socio-economic data, clinical history and sexual behaviour. Pelvic examinations were performed and cervical specimens were collected for detection of pathogens by PCR. The prevalence of infection was: 12.7% for Trichomonas vaginalis, 6.4% for Chlamydia trachomatis and 1.4% for Neisseria gonorrhoeae. There were no statistically significant associations between infections by any of the pathogens nor by any pathogen alone with any clinical variable, socio-demographic data or sexual behaviour. This study draws attention to the need for surveillance and possible need for screening for Chlamydia trachomatis, which often progresses asymptomatically. For the significant prevalence found, attention should also be given to asymptomatic infection by Trichomonas vaginalis, since this pathogen has recently been implicated as a risk factor for HIV infection.
Asunto(s)
Infecciones por Chlamydia/epidemiología , Chlamydia trachomatis/aislamiento & purificación , Gonorrea/epidemiología , Neisseria gonorrhoeae/aislamiento & purificación , Vaginitis por Trichomonas/epidemiología , Trichomonas vaginalis/aislamiento & purificación , Adulto , Brasil/epidemiología , Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , Femenino , Gonorrea/diagnóstico , Gonorrea/microbiología , Humanos , Neisseria gonorrhoeae/genética , Reacción en Cadena de la Polimerasa , Prevalencia , Atención Primaria de Salud , Factores de Riesgo , Conducta Sexual , Enfermedades de Transmisión Sexual/parasitología , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/genética , Adulto JovenRESUMEN
The aim of this study is to estimate the prevalence of Trichomonas vaginalis and Candida albicans in low-risk patients treated at a first level clinic (primary health care represents the first level of contact of individuals, families, and the community with the system national health). Using a cross-sectional study in patients treated in clinical laboratory of the Sanitary District no. 7 of the city of Orizaba during the months June-July, 252 urine samples were collected for the identification of T. vaginalis and C. albicans by PCR. Furthermore, we analyzed the sociodemographic characteristics of the studied population. We observed an overall prevalence of 23.41% (95% CI 22.10-24.72) for T. vaginalis and 38.88% (95% CI 37.73-40.03) for C. albicans. There was also presence of coinfection in 14.28% (95% CI 13.10-15.46), which was associated with the presence of pain. Most of the positive cases were observed in women house-maker (80%, 95% CI 50.36-48.98). The results of this study provide evidence that the majority of positive cases observed in the studied population are presented in an asymptomatic form and usually are not associated with any risk factor.
Asunto(s)
Candida albicans/aislamiento & purificación , Candidiasis/orina , Instituciones de Salud , Reacción en Cadena de la Polimerasa/métodos , Vaginitis por Trichomonas/orina , Trichomonas vaginalis/aislamiento & purificación , Adolescente , Adulto , Distribución por Edad , Anciano , Anciano de 80 o más Años , Candida albicans/genética , Candidiasis/diagnóstico , Candidiasis/microbiología , Femenino , Geografía , Humanos , México , Persona de Mediana Edad , Factores de Riesgo , Factores Socioeconómicos , Vaginitis por Trichomonas/diagnóstico , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/genética , Adulto JovenRESUMEN
The goal of this paper was to characterize a Trichomonas vaginalis cysteine proteinase (CP) legumain-1 (TvLEGU-1) and determine its potential role as a virulence factor during T. vaginalis infection. A 30-kDa band, which migrates in three protein spots (pI~6.3, ~6.5, and ~6.7) with a different type and level of phosphorylation, was identified as TvLEGU-1 by one- and two-dimensional Western blot (WB) assays, using a protease-rich trichomonad extract and polyclonal antibodies produced against the recombinant TvLEGU-1 (anti-TvLEGU-1r). Its identification was confirmed by mass spectrometry. Immunofluorescence, cell binding, and WB assays showed that TvLEGU-1 is upregulated by iron at the protein level, localized on the trichomonad surface and in lysosomes and Golgi complex, bound to the surface of HeLa cells, and was found in vaginal secretions. Additionally, the IgG and Fab fractions of the anti-TvLEGU-1r antibody inhibited trichomonal cytoadherence up to 45%. Moreover, the Aza-Peptidyl Michael Acceptor that inhibited legumain proteolytic activity in live parasites also reduced levels of trichomonal cytoadherence up to 80%. In conclusion, our data show that the proteolytic activity of TvLEGU-1 is necessary for trichomonal adherence. Thus, TvLEGU-1 is a novel virulence factor upregulated by iron. This is the first report that a legumain-like CP plays a role in a pathogen cytoadherence.
Asunto(s)
Cisteína Endopeptidasas/metabolismo , Proteasas de Cisteína/metabolismo , Proteínas Protozoarias/metabolismo , Trichomonas vaginalis/enzimología , Adhesión Celular , Proteasas de Cisteína/química , Proteasas de Cisteína/fisiología , Femenino , Aparato de Golgi/metabolismo , Células HeLa , Humanos , Inmunoglobulina G/inmunología , Hierro/metabolismo , Hígado/parasitología , Fosforilación , Proteínas Protozoarias/química , Proteínas Protozoarias/fisiología , Proteínas Recombinantes/metabolismo , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/patogenicidad , Vagina/parasitología , Factores de Virulencia/metabolismoRESUMEN
Women participating in studies in Brazil (n = 695) and South Africa (n = 230) performed rapid point-of-care tests for Trichomonas vaginalis on self-collected vaginal swabs. Using PCR as the gold standard, rapid self-testing achieved high specificity (99.1%; 95% confidence interval [CI], 98.2 to 99.6%) and moderate sensitivity (76.7%; 95% CI, 61.4 to 88.2%). These tests may be considered an alternative to syndromic management in resource-poor settings.
Asunto(s)
Parasitología/métodos , Sistemas de Atención de Punto , Autoexamen/métodos , Vaginitis por Trichomonas/diagnóstico , Trichomonas vaginalis/aislamiento & purificación , Adolescente , Adulto , Brasil , Femenino , Humanos , Sensibilidad y Especificidad , Sudáfrica , Vaginitis por Trichomonas/parasitología , Adulto JovenRESUMEN
This review focused on potential regulatory mechanisms of Trichomonas vaginalis virulence properties, cytoadherence, cytotoxicity, phagocytosis, hemolysis, induction of apoptosis, and immune evasion in response to environmental factors of the human urogenital tract, iron, zinc, and polyamines. Understanding the multifactorial nature of trichomonal pathogenesis and its regulation may help to unravel the survival strategies of trichomonads and to implement prevention policies, opportune diagnosis, and alternative treatments for control of trichomoniasis.