RESUMO
Trichomonas vaginalis is the causative agent of trichomoniasis, the most prevalent nonviral, neglected sexually transmitted disease worldwide. T. vaginalis has one of the largest degradomes among unicellular parasites. Cysteine peptidases (CPs) are the most abundant peptidases, constituting 50% of the degradome. Some CPs are virulence factors recognized by antibodies in trichomoniasis patient sera, and a few are found in vaginal secretions that show fluctuations in glucose concentrations during infection. The CPs of clan CD in T. vaginalis include 10 genes encoding legumain-like peptidases of the C13 family. TvLEGU-2 is one of them and has been identified in multiple proteomes, including the immunoproteome obtained with Tv (+) patient sera. Thus, our goals were to assess the effect of glucose on TvLEGU-2 expression, localization, and in vitro secretion and determine whether TvLEGU-2 is expressed during trichomonal infection. We performed qRT-PCR assays using parasites grown under different glucose conditions. We also generated a specific anti-TvLEGU-2 antibody against a synthetic peptide of the most divergent region of this CP and used it in Western blot (WB) and immunolocalization assays. Additionally, we cloned and expressed the tvlegu-2 gene (TVAG_385340), purified the recombinant TvLEGU-2 protein, and used it as an antigen for immunogenicity assays to test human sera from patients with vaginitis. Our results show that glucose does not affect tvlegu-2 expression but does affect localization in different parasite organelles, such as the plasma membrane, Golgi complex, hydrogenosomes, lysosomes, and secretion vesicles. TvLEGU-2 is secreted in vitro, is present in vaginal secretions, and is immunogenic in sera from Tv (+) patients, suggesting its relevance during trichomonal infection.
RESUMO
PROBLEM: The association of viruses with infertility remains incompletely evaluated. METHOD OF STUDY: Vaginal secretions from 46 women seeking treatment in the Center for Reproductive Medicine and Infertility at Weill Cornell Medicine were tested for viruses by metagenomic analysis by lab personnel blinded to all clinical data. RESULTS: Torquetenovirus (TTV) was identified in 16 women, alphapapillomavirus in seven women and most were positive for bacteriophages. Twelve of the subjects were fertile and sought to freeze their oocytes for future implantation. These women were all negative for TTV. In contrast, 16 of the 34 women (47.1%) being treated for infertility were TTV-positive (p = .0035). Evaluating the women by cause of infertility, five of nine women (55.6%) whose male partner had inadequate sperm parameters and six of 14 women (42.9%) with defective ovulation were TTV positive (p = .0062 and p = .0171, respectively, vs. the fertile women). Alphapapillomavirus was identified in one (8.3%) fertile woman, five (35.7%) women with ovulation deficiency, and one (11.1%) woman with male factor infertility. These differences were not statistically significant. There were no differences in bacteriophage families or the presence of Lactobacillus phages between fertile or infertile women or between different causes of infertility. There was a negative association between TTV detection and Lactobacillus crispatus dominance in the vaginal microbiota (p = .0184), but no association between TTV detection and the presence of alphapapillomavirus or Candida species. CONCLUSION: Detection of TTV in the vagina might be a biomarker for specific causes of infertility.
Assuntos
Infertilidade Feminina , Infertilidade Masculina , Lactobacillus crispatus , Torque teno virus , Masculino , Humanos , Feminino , Torque teno virus/genética , Sêmen , VaginaRESUMO
The microbial composition of the human vagina differs from that of all other mammals, likely as a consequence of the unique composition of vaginal constituents that promote the selective maintenance and proliferation of distinct bacterial species. Similarly, variations between individual healthy women in genetic, environmental, and medical variables also modify the vaginal lumen composition. The direction and magnitude of immune reactions to microorganisms present in the vagina, responses to stress and non-infectious stimuli, coupled with medical and pregnancy history and environmental exposures, can greatly differ between women. Adaptations to both internal and external pressures will determine the ability of select resident vaginal bacteria to numerically dominate and, therefore, the definition of a "normal" vaginal microbiota will substantially differ between individual healthy women.
Assuntos
Microbiota , Vagina , Gravidez , Animais , Feminino , Humanos , Vagina/microbiologia , Microbiota/fisiologia , Reprodução , Bactérias , RNA Ribossômico 16S , Mamíferos/genéticaRESUMO
Trichomonas vaginalis (Tv) induces host cell damage through cysteine proteinases (CPs) modulated by iron. An immunoproteomic analysis showed that trichomoniasis patient sera recognize various CPs, also some of them are present in vaginal washes (VWs). Thus, the goal of this work was to determine whether TvCP2 is expressed during infection and to assess the effect of iron on TvCP2 expression, localization and contribution to in vitro cellular damage. Western-blotting (WB) assays using TvCP2r and vaginitis patient serum samples showed that 6/9 Tv (+) but none of the Tv (-) patient sera recognized TvCP2r. WB using an anti-TvCP2r antibody and VWs from the same patients showed that in all of the Tv (+) but none of the Tv (-) VWs, the anti-TvCP2r antibody detected a 27 kDa protein band that corresponded to the mature TvCP2, which was confirmed by mass spectrometry analysis. Iron decreased the amount of TvCP2 mRNA and the protein localized on the parasite surface and cytoplasmic vesicles concomitant with the cytotoxic effect of TvCP2 on HeLa cells. Parasites pretreated with the anti-TvCP2r antibody also showed reduced levels of cytotoxicity and apoptosis induction in HeLa cell monolayers. In conclusion, these results show that TvCP2 is expressed during trichomonal infection and plays an important role in the in vitro HeLa cell cytotoxic damage under iron-restricted conditions.
Assuntos
Cisteína Proteases/metabolismo , Ferro/administração & dosagem , Proteínas de Protozoários/metabolismo , Trichomonas vaginalis/efeitos dos fármacos , Vagina/parasitologia , Secreções Corporais/parasitologia , Feminino , Humanos , Trichomonas vaginalis/enzimologiaRESUMO
Zika virus, an arthropod-borne flavivirus pathogen in humans, is unusual because it can be sexually transmitted and can be shed for prolonged periods in semen. We report viral shedding in vaginal secretions for up to 6 months, indicating the potential for sexual and vertical transmission by infected women.
Assuntos
RNA Viral/isolamento & purificação , Eliminação de Partículas Virais , Infecção por Zika virus/transmissão , Zika virus/isolamento & purificação , Feminino , Humanos , Transmissão Vertical de Doenças Infecciosas , Nicarágua , Vagina/virologia , Infecção por Zika virus/virologiaRESUMO
Infection with Zika virus is an emerging public health crisis. We observed prolonged detection of virus RNA in vaginal mucosal swab specimens and whole blood for a US traveler with acute Zika virus infection who had visited Honduras. These findings advance understanding of Zika virus infection and provide data for additional testing strategies.
Assuntos
RNA Viral/sangue , Vagina/virologia , Infecção por Zika virus/virologia , Adulto , Animais , Chlorocebus aethiops , Meios de Cultivo Condicionados/química , Feminino , Honduras , Humanos , RNA Viral/urina , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saliva/virologia , Fatores de Tempo , Viagem , Estados Unidos , Vagina/metabolismo , Células Vero , Zika virus/genética , Zika virus/crescimento & desenvolvimento , Infecção por Zika virus/sangue , Infecção por Zika virus/fisiopatologia , Infecção por Zika virus/urinaRESUMO
The puerperium is the period between birth and complete uterine involution, which is around 40 days postpartum (dpp) in cattle. This study evaluated the dynamics of uterine involution in 19 crossbred dairy cows, who had given one or more births. The evaluation was done by observing the vital characteristics, body score, ultrasonographic and gynecological examinations, and the odor and characteristics of vaginal secretions. These evaluations were done during eight different moments of puerperium: day 0, 3, 7, 10, 14, 21, 28, and 43. The results from this study demonstrated that there are specific differences relative to uterine involution in crossbred dairy cows at 21 and 28 dpp. Additionally, these results suggest that the diagnosis and treatment of clinical endometritis normally done by evaluating the characteristics and the odor of vaginal secretions should be revised, since spontaneous recuperation of endometritis occurred at 43 dpp.(AU)
Puerpério é o período compreendido entre o parto e o momento em que ocorre a completa involução uterina em torno de 40 dias pós-parto (dpp). Com o objetivo de avaliar a dinâmica de involução uterina em vacas leiteiras foram examinadas 19 vacas, mestiças, com dois ou mais partos em oito momentos no puerpério: dia do parto, dia 3, 7, 10, 14, 21, 28 e 43 dpp, onde se verificou os parâmetros vitais, escore de condição corporal, exames ginecológico e ultrassonográfico, características e odor de secreção vaginal. Concluiu-se que existem características particulares referentes à involução uterina nas vacas mestiças leiteiras e que o diagnóstico e tratamento da endometrite clínica, através da avaliação aos 21 e 28 dpp das características e odor de secreção vaginal devem ser revistos, pois se verificou cura espontânea desta infecção aos 43 dpp. Palavras-chave: puerpério, secreção vaginal, raça mestiça, exame ginecológico, ultrassonografia transretal.(AU)