Trichomonas vaginalis extracellular vesicles are internalized by host cells using proteoglycans and caveolin-dependent endocytosis.

Trichomonas vaginalis, a human-infective parasite, causes the most prevalent nonviral sexually transmitted infection worldwide. This pathogen secretes extracellular vesicles (EVs) that mediate its interaction with host cells. Here, we have developed assays to study the interface between parasite EVs and mammalian host cells and to quantify EV internalization by mammalian cells. We show that T. vaginalis EVs interact with glycosaminoglycans on the surface of host cells and specifically bind to heparan sulfate (HS) present on host cell surface proteoglycans. Moreover, competition assays using HS or removal of HS from the host cell surface strongly inhibit EV uptake, directly demonstrating that HS proteoglycans facilitate EV internalization. We identified an abundant protein on the surface of T. vaginalis EVs, 4-α-glucanotransferase (Tv4AGT), and show using isothermal titration calorimetry that this protein binds HS. Tv4AGT also competitively inhibits EV uptake, defining it as an EV ligand critical for EV internalization. Finally, we demonstrate that T. vaginalis EV uptake is dependent on host cell cholesterol and caveolin-1 and that internalization proceeds via clathrin-independent, lipid raft-mediated endocytosis. These studies reveal mechanisms used to drive host:pathogen interactions and further our understanding of how EVs are internalized by target cells to allow cross-talk between different cell types.

Protocolo Sociedad Española de Ginecología y Obstetricia de diagnóstico y tratamiento de las infecciones vulvovaginales. Protocolo actualizado en 2012 / The protocol of the Spanish Society of Obstetrics and Gynecology for the diagnosis and treatment of vulvovaginal infection. Update 2012

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Vaginitis.

Bacterial vaginosis, trichomoniasis, and vulvovaginal candidiasis are the most common infectious causes of vaginitis. Bacterial vaginosis occurs when the normal lactobacilli of the vagina are replaced by mostly anaerobic bacteria. Diagnosis is commonly made using the Amsel criteria, which include vaginal pH greater than 4.5, positive whiff test, milky discharge, and the presence of clue cells on microscopic examination of vaginal fluid. Oral and topical clindamycin and metronidazole are equally effective at eradicating bacterial vaginosis. Symptoms and signs of trichomoniasis are not specific; diagnosis by microscopy is more reliable. Features of trichomoniasis are trichomonads seen microscopically in saline, more leukocytes than epithelial cells, positive whiff test, and vaginal pH greater than 5.4. Any nitroimidazole drug (e.g., metronidazole) given orally as a single dose or over a longer period resolves 90 percent of trichomoniasis cases. Sex partners should be treated simultaneously. Most patients with vulvovaginal candidiasis are diagnosed by the presence of vulvar inflammation plus vaginal discharge or with microscopic examination of vaginal secretions in 10 percent potassium hydroxide solution. Vaginal pH is usually normal (4.0 to 4.5). Vulvovaginal candidiasis should be treated with one of many topical or oral antifungals, which appear to be equally effective. Rapid point-of-care tests are available to aid in accurate diagnosis of infectious vaginitis. Atrophic vaginitis, a form of vaginitis caused by estrogen deficiency, produces symptoms of vaginal dryness, itching, irritation, discharge, and dyspareunia. Both systemic and topical estrogen treatments are effective. Allergic and irritant contact forms of vaginitis can also occur.

Trichomoniasis: evaluation to execution.

Trichomoniasis is the most common sexually transmitted disease, caused by a motile flagellate non-invasive parasitic protozoan, Trichomonas vaginalis (T. vaginalis). More than 160 million people worldwide are annually infected by this protozoan. T. vaginalis occupies an extracellular niche in the complex human genito-urinary environment (vagina, cervix, penis, prostate gland, and urethra) to survive, multiply and evade host defenses. T. vaginalis (strain G3) has a ∼160 megabase genome with 60,000 genes, the largest number of genes ever identified in protozoans. The T. vaginalis genome is a highly conserved gene family that encodes a massive proteome with one of the largest coding (expressing ∼4000 genes) capacities in the trophozoite stage, and helps T. vaginalis to adapt and survive in diverse environment. Based on recent developments in the field, we review T. vaginalis structure, patho-mechanisms, parasitic virulence, and advances in diagnosis and therapeutics.

Antitrichomonas IgG, IgM, IgA, and IgG subclass responses in human intravaginal trichomoniasis.

Trichomoniasis, caused by the protozoan parasite Trichomonas vaginalis, is a major nonviral sexually transmitted disease. Clinical spectrum varies from an asymptomatic state to mild, moderate, or severe symptoms. However, the exact factors leading to the variations in symptoms have not been well elucidated. Host's immune response to the parasite may be playing a role in varied symptomatology. The present study reports antitrichomonas IgM, IgA, IgG and its subclasses in doubling dilutions of serum and diluted vaginal washes of six T. vaginalis-infected symptomatic and four T. vaginalis-infected asymptomatic women and uninfected controls by enzyme-linked immunosorbent assay (ELISA). No significant difference was observed in serum IgG ELISA absorbance values from symptomatic compared to asymptomatic subjects (p > 0.05) while a significant difference (p < 0.05) was noted in serum IgM in all the tested dilutions and IgA up to a dilution of 400. This is the first report of the detection of specific IgG subclass response in T. vaginalis-infected female patients, and quantitative analysis of the antibody responses indicated that the production of local IgG particularly IgG1 in vaginal secretions may be playing a significant role in establishing symptomatic infection. The interesting observation of the present study is that the specific IgM was detected in 2 (33.3%) symptomatic and T. vaginalis-infected patients in > or =800 dilutions and in 1 (16.6%) up to 200 dilutions in serum, while it was not detectable in the vaginal secretions of symptomatic patients or in the serum and vaginal secretions of asymptomatic T. vaginalis-infected patients.

Prevalence of sexually transmitted infections among sexually active Jordanian females.

OBJECTIVES: To determine the prevalence of the most important sexually transmitted infections among women of child bearing age in Jordan. GOAL: To assess the need for screening programs to detect sexually transmitted infections. STUDY DESIGN: This is a cross-sectional study wherein consecutive symptomatic and asymptomatic women presenting to gynecology and family planning clinics from different areas in Jordan were tested for reproductive tract infections caused by Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum, and Trichomonas vaginalis. RESULTS: The prevalence of C. trachomatis infection was 0.6% and 0.5%, among symptomatic and asymptomatic women respectively, that of N. gonorrhoeae was 0.9% and 2.2%, that of T. pallidum 0.0% and 0.0%, and that of Tr. vaginalis was 0.7% and 0.5%. These prevalence rates did not differ significantly between symptomatic and asymptomatic women. CONCLUSIONS: Based on the low prevalence of sexually transmitted infections detected in this study among Jordanian women, the need for screening programs for such infections is questioned.

Altered expression of adhesion molecules in inflammatory cervical smears.

OBJECTIVE: The aim of the present study was to evaluate the expression of pan-cadherin and beta-catenin in cervical smears with various types of infectious agents. PATIENTS AND METHODS: Cervical smears obtained from 53 women, aged 21-65 years, with a diagnosis of specific inflammation were examined in our study. Eighteen subjects were infected by Candida albicans, 18 by Gardnerella vaginalis, nine by Bacteroides spp. and eight by Chlamydia trachomatis. All infectious agents found in the smears were at the same time confirmed by the microbiological laboratory methods. We performed a biotin-streptavidin-peroxidase immunocytochemical method using anti-beta-catenin (Clone 12F7) and anti-pan-cadherin (pan, polyclonal) antibodies. RESULTS: Aberrant expression of pan-cadherin was found in the cytoplasmic membrane of glandular, metaplastic, superficial and intermediate squamous cells in all types of infections. With regard to beta-catenin, this was expressed in majority (90%) of glandular and metaplastic cells in all types of infections and in a small proportion (15%) of superficial and intermediate squamous cells in infections caused by C. albicans and G. vaginalis. CONCLUSION: Our data show that infectious agents may cause alterations in the expression and distribution of these adhesive molecules, which can be recognized in cervical smears. Additional studies in larger sets of patients should help clarify this issue further.

Cysteine proteinase 30 (CP30) and antibody response to CP30 in serum and vaginal washes of symptomatic and asymptomatic Trichomonas vaginalis-infected women.

Infection with Trichomonas vaginalis may be asymptomatic or with symptoms suggestive of vaginitis. Because cysteine proteinase 30 (CP30) of T. vaginalis is known to be a virulence marker that plays a role in cytoadherence, the aim of this study was to analyse the presence of CP30 and antibody to CP30 in clinical samples of symptomatic and asymptomatic infected women. CP30 was detected in all the serum and vaginal washes (VWs) of symptomatic women and in 65% of the serum and 80% of the VWs of asymptomatic women. This suggested that the majority of asymptomatic women also exhibit CP30 in the serum and VWs. Antibody to CP30 was detected in all the serum samples of symptomatic and asymptomatic women and in the VWs of only 54.5% of the symptomatic and 35% of the asymptomatic women. Antibody to CP30 was also detected in 3/20 of the serum samples and in none of the VWs from uninfected women. Significantly higher amounts of antibody (mean OD values) were observed in serum and VWs of symptomatic as compared to asymptomatic and healthy women (P<0.001). These results indicate that besides CP30, other factors may also be playing a role in leading to symptomatic infection, because CP30 was detected in clinical samples from all the symptomatic and the majority of the asymptomatic women. Although anti-CP30 antibodies do not appear to be protective, detection of antibody to CP30 antigen in serum samples may be used as a diagnostic tool.

Oral and vulvar changes in pregnancy.

The endocrinologic, immunological, and vascular changes that occur during pregnancy are far-reaching. These systemic factors produce profound local impact on the physiology and pathology of the oral cavity, vagina, and vulva. Physiological changes can be expected and tolerable or of such severity that they are viewed as pathological. Existing disease can be exacerbated and dermatoses specific to pregnancy can erupt. Each of these conditions can pose potentially challenging treatment considerations.

Trichomonas vaginalis in a Sheffield genitourinary medicine department.

The case-notes of all patients who were diagnosed with a first episode of Trichomonas vaginalis (TV) between 1 October 2002 and 30 September 2003 were reviewed. A total of 78 patients were suitable for inclusion in the study. Analysis of their notes revealed that, although the majority of patients presented with symptoms, 15% (n=12) of cases were asymptomatic. A raised vaginal pH was found in 94% (n=47) of the patients in whom it was measured. In all, 97% (n=76) of patients received treatment in accordance with UK national guidelines and, in those tested, initial treatment was found to be 95% (n=57) successful. Treatment of at least one contact could only be confirmed in 27% (n=21) of cases. The implications for future management of TV are discussed.

Trichomonas vaginalis: intrastrain polymorphisms within the ribosomal intergenic spacer do not correlate with clinical presentation.

Trichomoniasis presents a broad spectrum of clinical patterns ranging from asymptomatic to severe vaginitis and cervicitis. Despite its importance, very little is known about the genetic relatedness of its causative agent, Trichomonas vaginalis, and the clinical phenotypes. To address this question, analysis of restriction length polymorphism (RFLP) within the intergenic spacer of the ribosomal DNA (IGS) from 60 clinically defined isolates of T. vaginalis was performed. This is the first description of the IGS polymorphism of T. vaginalis. As expected, a considerable number of patients were asymptomatic (28%) while only 12% presented both leukorrhea and macular colpitis, the most evident symptoms of trichomoniasis. The IGS-RFLP with the use of eight restriction enzymes showed absence of correlation between the genetic relatedness of the isolates and symptomatology. Further studies are necessary to evaluate the importance of the IGS polymorphism to the parasite virulence and clinical phenotype.

Trichomonas vaginalis: random amplified polymorphic DNA analysis of isolates from symptomatic and asymptomatic women in India.

Trichomonas vaginalis, the causative agent for human trichomoniasis, is a protozoan parasite. Trichomoniasis is the most common non-viral sexually transmitted disease. The infection in women may be asymptomatic or may lead to severe vaginitis, cervicitis and severe sequelae. Despite its high prevalence, the genetic variability and factors leading to symptomatic infection have been poorly understood. One thousand women in childbearing age group were screened for the presence of T vaginalis. Thirty-eight women were found positive for T vaginalis and out of these 22 (57.9%) were having symptomatic infection and 16 (42%) were asymptomatic. Fresh isolates from 15 symptomatic and 15 asymptomatic women were axenised and subjected to random amplified polymorphic DNA (RAPD) analysis with the use of five different random primers (OPD 1-OPD 5). The isolates with similar banding pattern were assigned as a single type. OPD 3 indicated least (nine types) while OPD 4 indicated highest typing (18 types) ability. Phylogenetic analysis using RAPD distance software indicated two distinct lineages; upper branch consisting of only seven symptomatic isolates while lower branch consisting of all the 15 asymptomatic isolates the other eight symptomatic isolates were recorded in separate cluster. The study indicated that RAPD technique might be helpful to delineate the pathogenic mechanism(s) for its virulence; however, further studies on large number of isolates are desired to elucidate the findings.

Subtipos de virus del papiloma humano y lesiones intraepiteliales e invasoras de cérvix uterino en mujeres de la provincia de Ciudad Real / Subtypes of human Papillomavirus and intraepithelial invasive lesions of the uterine cervix in women in the area of Ciudad Real

Introducción: El papel oncogénico dei virus dei papíloma humano (VPH) en las lesiones intraepiteliales de cérvix uterino se ha descrito en numerosas publicaciones. Objetivo: Conocer la prevalencia y los subtipos de infección por VPH en pacientes con lesiones citológicas. Material y métodos: Durante dos años se han diagnosticado las triples tomas citológicas de 6300 mujeres. En todas las lesiones intraepiteliales y carcinomas invasores se determinó la presencia de ADN vírico dei VPH mediante reacción en cadena de polimerasa (RCP). Resultados: El VPH fue positivo en el 20 por ciento de las lesiones de bajo grado, en el 73 por ciento de las de alto grado y en el 50 por ciento de los carcinomas invasores. El subtipo más frecuente en lesiones de bajo grado fue "no upado". El subtípo 16 se asoció a lesiones de alto grado y a carcinoma invasor. En cuatro ocasiones se aisló más de un subtípo de VPH. Conclusiones: Los subtipos de VPH aislados son similares a los descritos en nuestro medio (AU)

Clinical and microbiological correlates of vaginal trichomoniasis during pregnancy. The Vaginal Infections and Prematurity Study Group.

Colonization with Trichomonas vaginalis is a possible cause of poor pregnancy outcome. To facilitate the diagnosis of this condition during pregnancy, we conducted a prospective, multicenter study of 13,816 gravid women who were between 23rd and 26th week of gestation. Findings significantly associated with T. vaginalis colonization included a yellow, green, or bloody discharge from the vagina or cervix; abnormal odor after KOH was added to a vaginal specimen; a vaginal pH of > 5.0; and cervical friability. The amount of vaginal discharge and abnormal consistency of the discharge were also associated with T. vaginalis colonization. These findings (except for cervical bleeding and odor after the addition of KOH to a vaginal specimen, which may be influenced by the presence of other flora) are consistent with those reported elsewhere. The clinical usefulness of these features is minimal, and it is more significant that other microorganisms are makers for trichomoniasis; therefore, controlling for other flora is important in the investigation of T. vaginalis colonization.

Vaginitis: diagnosis and management.

The various conditions that give rise to vaginitis include specific and nonspecific entities, such as candidiasis, trichomoniasis, bacterial vaginosis, group B streptococcal vaginitis, purulent vaginitis, volvodynia, and vestibulitis. The patient with chronic vaginitis usually develops this condition because of a misdiagnosis. It is critical that patients who have chronic vaginitis be thoroughly evaluated to determine if there is a specific etiology and whether their condition is recurrent or persistent, or is a reinfection. This also must include obtaining a detailed history, beginning with the patient's best recollection of when she felt perfectly normal. The physician must have an understanding of a healthy vaginal ecosystem and what mechanisms are in place to maintain the equilibrium. The vaginal ecosystem is a complex system of micro-organisms interacting with host factors to maintain its equilibrium. The endogenous microflora consists of a variety of bacteria, which include aerobic, facultative and obligate anaerobic bacteria. These organisms exist in a commensal, synergistic or antagonistic relationship. Therefore, it is important to understand what factors control the delicate equilibrium of the vaginal ecosystem, and which factors, both endogenous and exogenous, can disrupt this system. It is also important for the physician to understand that when a patient has symptoms of vaginitis it is not always due to an infectious etiology. There are situations in which an inflammatory reaction occurs but the specific etiology may not be determined. Thus, it is important that the physician not rush through the history or the examination.

Vaginal candidiasis and other types of vaginitis.

AIDS: The causes, diagnosis, treatment options, and recent research findings of vaginal infections in HIV-infected women are discussed. Three common types of vaginitis are addressed: vaginal candidiasis, bacterial vaginosis, and trichomoniasis. The prevalence of candidiasis is an important indicator of HIV disease severity and data is provided that shows its frequency, compared to other genital infections, in HIV-positive women.^ieng

The relationship of experimental pathogenicity in vivo with in vitro cytoadherence and cytotoxicity of 6 different isolates of Trichomonas vaginalis.

The experimental pathogenic effects in vivo and in vitro of 6 different isolates of Trichomonas vaginalis were studied following their inoculation into NMRI mice and on to adherent cultures of HeLa cells. Contact between the parasite and the adherent monolayer of cells was necessary to induce the monolayer to detach. The strains which were more virulent to mice also showed a greater weighted index of adherence; the weighted index of cytotoxicity in vitro did not, on the other hand, correlate with experimental pathology in vivo.