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1.
Anal Chem ; 93(3): 1544-1552, 2021 01 26.
Artículo en Inglés | MEDLINE | ID: mdl-33378175

RESUMEN

Mapping highly complicated disulfide linkages and free thiols via liquid chromatography-tandem mass spectrometry (LC-MS2) is challenging because of the difficulties in optimizing sample preparation to acquire critical MS data and detecting mispairings. Herein, we report a highly efficient and comprehensive workflow using an on-line UV-induced precolumn reduction tandem mass spectrometry (UV-LC-MS2) coupled with two-stage data analysis and spiked control. UV-LC-MS2 features a gradient run of acetonitrile containing a tunable percentage of photoinitiators (acetone/alcohol) that drives the sample to the MS through a UV-flow cell and reverse phase column to separate UV-induced products for subsequent fragmentation via low energy collision-induced dissociation. This allowed the alkylated thiol-containing and UV-reduced cysteine-containing peptides to be identified by a nontargeted database search. Expected or unexpected disulfide/thiol mapping was then carried out based on the search results, and data were derived from partially reduced species by photochemical reaction. Complete assignments of native and scrambled disulfide linkages of insulin, α-lactalbumin, and bovine serum albumin (BSA) as well as the free C34-BSA were demonstrated using none or single enzyme digestion. This workflow was applied to characterize unknown disulfide/thiol patterns of the recombinant cyclophilin 1 monomer (rTvCyP1 mono) from the human pathogen Trichomonas vaginalis. α-Lactalbumin was judiciously chosen as a spiked control to minimize mispairings due to sample preparation. rTvCyP1 was determined to contain a high percentage of thiol (>80%). The rest of rTvCyP1 mono were identified to contain two disulfide/thiol patterns, of which C41-C169 linkage was confirmed to exist as C53-C181 in rTvCyP2, a homologue of rTvCyP1. This platform identifies heterogeneous protein disulfide/thiol patterns in a de-novo fashion with artifact control, opening up an opportunity to characterize crude proteins for many applications.


Asunto(s)
Ciclofilinas/análisis , Disulfuros/química , Lactalbúmina/química , Compuestos de Sulfhidrilo/química , Trichomonas vaginalis/química , Rayos Ultravioleta , Humanos , Oxidación-Reducción , Proteínas Recombinantes/análisis , Espectrometría de Masas en Tándem
2.
BMC Infect Dis ; 20(1): 642, 2020 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-32873233

RESUMEN

BACKGROUND: Evidence suggested that vaginal microbiome played a functional role in the progression of cervical lesions in female infected by HPV. This study aimed at evaluating the influence of common vaginal infection on the carcinogenicity of high risk HPV (hr-HPV). METHODS: From January 15, 2017 to December 31, 2017, 310,545 female aged at least 30 years old had been recruited for cervical cancer screening from 9 clinical research centers in Central China. All the recruited participants received hr-HPV genotyping for cervical cancer screening and vaginal microenvironment test by a high vaginal swab. Colposcopy-directed biopsy was recommended for female who were infected with HPV 16 and HPV 18, and other positive hr-HPV types through test had undertaken triage using liquid-based cytology, cases with the results ≥ ASCUS among them were referred to colposcopy directly, and cervical tissues were taken for pathology examination to make clear the presence or absence of other cervical lesions. RESULTS: Among 310,545 female, 6067 (1.95%) were tested with positive HPV 16 and HPV 18, 18,297 (5.89%) were tested with other positive hr-HPV genotypes, cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3 and invasive cervical cancer (ICC) were detected in 861 cases, 377 cases, 423 cases, and 77 cases, respectively. Candida albicans and Gardnerella were not associated with the detection of cervical lesions. Positive trichomonas vaginitis (TV) was correlated with hr-HPV infection (p < 0.0001). Co-infection with TV increased the risk of CIN 1 among female infected with hr-HPV (OR 1.18, 95% CI: 1.42-2.31). Co-infection with TV increased the risk of CIN 2-3 among female infected with HPV 16 (OR 1.71, 95% CI: 1.16-2.53). CONCLUSIONS: Co-infection of TV and HPV 16 is a significant factor for the detection of cervical lesions.


Asunto(s)
Neoplasia Intraepitelial Cervical/epidemiología , Coinfección/complicaciones , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Infecciones por Papillomavirus/complicaciones , Vaginitis por Trichomonas/complicaciones , Trichomonas vaginalis/aislamiento & purificación , Neoplasias del Cuello Uterino/epidemiología , Adulto , Neoplasia Intraepitelial Cervical/diagnóstico , Neoplasia Intraepitelial Cervical/virología , China/epidemiología , Coinfección/diagnóstico , Colposcopía , Estudios Transversales , Citodiagnóstico , Detección Precoz del Cáncer/métodos , Femenino , Genotipo , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/diagnóstico , Infecciones por Papillomavirus/patología , Infecciones por Papillomavirus/virología , Factores de Riesgo , Vaginitis por Trichomonas/diagnóstico , Vaginitis por Trichomonas/parasitología , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/virología
3.
PLoS One ; 15(8): e0237982, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32817700

RESUMEN

Some organisms, like Trichomonas vaginalis, contain mitochondria-related hydrogen-producing organelles, called hydrogenosomes. The protein targeting into these organelles is proposed to be similar to the well-studied mitochondria import. Indeed, S. cerevisiae mitochondria and T. vaginalis hydrogenosomes share some components of protein import complexes. However, it is still unknown whether targeting signals directing substrate proteins to hydrogenosomes can support in other eukaryotes specific mitochondrial localization. To address this issue, we investigated the intracellular localization of three hydrogenosomal tail-anchored proteins expressed in yeast cells. We observed that these proteins were targeted to both mitochondria and ER with a variable dependency on the mitochondrial MIM complex. Our results suggest that the targeting signal of TA proteins are only partially conserved between hydrogenosomes and yeast mitochondria.


Asunto(s)
Retículo Endoplásmico/metabolismo , Regulación Fúngica de la Expresión Génica , Mitocondrias/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/citología , Trichomonas vaginalis/citología , Transporte de Proteínas , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Trichomonas vaginalis/genética , Trichomonas vaginalis/metabolismo
4.
Epidemiol Mikrobiol Imunol ; 69(2): 96-99, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32819109

RESUMEN

Neonatal pneumonia is mostly bacterial and other etiology is considered less frequently. We report a case of newborn whose neonatal pneumonia has not improved, despite the aggressive ventilation regime and empiric antibiotic therapy. A special sample from the respiratory tract was collected for PCR examination. The test confirmed the presence of Trichomonas vaginalis. Antibiotic therapy was extended to include metronidazole. Targeted antibiotic therapy, which lasted for 28 days, improved the condition and the patient was discharged in a stabilized condition to home care on the 44th day of life. We demonstrate the need to consider atypical pathogens in the case of infections that do not respond to conventional therapy. The multiplex real-time PCR technique was used to detect the DNA of the pathogen. Targeted antibiotic therapy is the result of pathogen identification.


Asunto(s)
Neumonía , Tricomoniasis , Trichomonas vaginalis , Humanos , Recién Nacido , Metronidazol/uso terapéutico , Reacción en Cadena de la Polimerasa Multiplex , Tricomoniasis/diagnóstico , Tricomoniasis/tratamiento farmacológico , Trichomonas vaginalis/genética
5.
Korean J Parasitol ; 58(3): 217-227, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32615735

RESUMEN

Trichomonas vaginalis causes inflammation of the prostate and has been detected in tissues of prostate cancers (PCa), prostatitis and benign prostatic hyperplasia. Obesity is a risk factor for PCa and causes a chronic subclinical inflammation. This chronic inflammation further exacerbates adipose tissue inflammation as results of migration and activation of macrophages. Macrophages are the most abundant immune cells in the PCa microenvironment. M2 macrophages, known as Tumor-Associated Macrophages, are involved in increasing cancer malignancy. In this study, conditioned medium (TCM) of PCa cells infected with live trichomonads contained chemokines that stimulated migration of the mouse preadipocytes (3T3-L1 cells). Conditioned medium of adipocytes incubated with TCM (ATCM) contained Th2 cytokines (IL-4, IL-13). Macrophage migration was stimulated by ATCM. In macrophages treated with ATCM, expression of M2 markers increased, while M1 markers decreased. Therefore, it is suggested that ATCM induces polarization of M0 to M2 macrophages. In addition, conditioned medium from the macrophages incubated with ATCM stimulates the proliferation and invasiveness of PCa. Our findings suggest that interaction between inflamed PCa treated with T. vaginalis and adipocytes causes M2 macrophage polarization, so contributing to the progression of PCa.


Asunto(s)
Adipocitos/inmunología , Macrófagos/inmunología , Neoplasias de la Próstata/etiología , Neoplasias de la Próstata/parasitología , Trichomonas vaginalis , Animales , Comunicación Celular/inmunología , Línea Celular Tumoral , Quimiocinas/inmunología , Inflamación , Masculino , Ratones , Obesidad , Neoplasias de la Próstata/patología , Factores de Riesgo
6.
PLoS One ; 15(6): e0234704, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32544192

RESUMEN

OBJECTIVE: Trichomonas vaginalis (TV) infection is common, curable, and associated with significant reproductive morbidity and risk for HIV infection. This analysis updates estimates of the prevalence of asymptomatic TV infection, and its associated risk factors, in the non-institutionalized U.S. population. METHODS: We analyzed data from 4057 individuals who participated in the National Health and Nutrition Examination Survey (NHANES) 2013-2014 data collection cycle. Participant interviews ascertained demographic characteristics, self-reported tobacco use, and sexual history. Self-collected urine specimens from participants aged 18 to 59 years were tested for TV infection using the Gen-Probe Aptima TV assay. Cotinine was assayed from serum to provide a biomarker of recent tobacco exposure. Weighted percentages are provided to account for unequal selection probabilities among participants and adjustments for non-response. RESULTS: Our sample included 1942 men (49.2%, 95% Confidence Interval [CI] 48.0-50.5) and 2115 women (50.8%, 95%CI 49.5-52.0). The infection prevalence among men was 0.5% (n = 16; 95%CI 0.2-1.0) and 1.8% (n = 55; 95%CI 1.1-3.1) in women. After controlling for participant characteristics associated with TV infection, females had a 5.2-fold increased odds of being infected compared to men (adjusted odds ratio (aOR) 5.2, 95% CI 2.4-11.4). Non-Hispanic blacks were more likely to be infected compared to non-Hispanic whites (aOR 11.2, 95% CI 4.6-27.2). Individuals below the federal poverty level were more likely to be infected compared to those earning >3 times the federal poverty level (aOR 6.7, 95% CI 1.7-26.6), and active smokers were more likely to be infected compared to participants with no nicotine exposure (aOR 8.7, 95% CI 4.1-18.2). CONCLUSION: Trichomonas vaginalis infection continues to be relatively common, especially in women, smokers, non-Hispanic blacks, and in groups of lower socioeconomic status. Identifying the demographic characteristics of populations in the United States disproportionately affected by TV could impact screening and treatment of this infection in clinical practice. Further research on whether screening and treating for asymptomatic TV infection in high-risk populations improves risk for reproductive morbidity and HIV infection is warranted.


Asunto(s)
Tricomoniasis/diagnóstico , Trichomonas vaginalis/aislamiento & purificación , Adolescente , Adulto , Cotinina/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Encuestas Nutricionales , Oportunidad Relativa , Pobreza , Prevalencia , Factores de Riesgo , Conducta Sexual , Fumar , Tricomoniasis/epidemiología , Tricomoniasis/etnología , Tricomoniasis/parasitología , Estados Unidos/epidemiología , Adulto Joven
7.
Parasitol Res ; 119(8): 2649-2657, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32583161

RESUMEN

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.


Asunto(s)
Tricomoniasis/complicaciones , Trichomonas vaginalis/patogenicidad , Enfermedades del Cuello del Útero/patología , Enfermedades del Cuello del Útero/parasitología , Adulto , Estudios de Casos y Controles , Depresión/complicaciones , Depresión/epidemiología , Femenino , Humanos , Persona de Mediana Edad , Oportunidad Relativa , Taiwán/epidemiología , Tricomoniasis/epidemiología , Enfermedades del Cuello del Útero/epidemiología , Enfermedades del Cuello del Útero/psicología , Neoplasias del Cuello Uterino/epidemiología , Neoplasias del Cuello Uterino/parasitología , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/psicología
8.
Parasitol Res ; 119(8): 2587-2595, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32524267

RESUMEN

Lycorine is an Amaryllidaceae alkaloid that presents anti-Trichomonas vaginalis activity. T. vaginalis causes trichomoniasis, the most common non-viral sexually transmitted infection. The modulation of T. vaginalis purinergic signaling through the ectonucleotidases, nucleoside triphosphate diphosphohydrolase (NTPDase), and ecto-5'-nucleotidase represents new targets for combating the parasite. With this knowledge, the aim of this study was to investigate whether NTPDase and ecto-5'-nucleotidase inhibition by lycorine could lead to extracellular ATP accumulation. Moreover, the lycorine effect on the reactive oxygen species (ROS) production by neutrophils and parasites was evaluated as well as the alkaloid toxicity. The metabolism of purines was assessed by HPLC. ROS production was measured by flow cytometry. Cytotoxicity against epithelial vaginal cells and fibroblasts was tested, as well as the hemolytic effect of lycorine and its in vivo toxicity in Galleria mellonella larvae. Our findings showed that lycorine caused ATP accumulation due to NTPDase inhibition. The alkaloid did not affect the ROS production by T. vaginalis; however, it increased ROS levels in neutrophils incubated with lycorine-treated trophozoites. Lycorine was cytotoxic against vaginal epithelial cells and fibroblasts; conversely, it was not hemolytic neither exhibited toxicity against the in vivo model of G. mellonella larvae. Overall, besides having anti-T. vaginalis activity, lycorine modulates ectonucleotidases and stimulates neutrophils to secrete ROS. This mechanism of action exerted by the alkaloid could enhance the susceptibility of T. vaginalis to host immune cell, contributing to protozoan clearance.


Asunto(s)
Alcaloides de Amaryllidaceae/farmacología , Amaryllidaceae/química , Antiprotozoarios/farmacología , Neutrófilos/metabolismo , Nucleósido-Trifosfatasa/antagonistas & inhibidores , Fenantridinas/farmacología , Extractos Vegetales/farmacología , Proteínas Protozoarias/antagonistas & inhibidores , Tricomoniasis/metabolismo , Trichomonas vaginalis/enzimología , 5'-Nucleotidasa/antagonistas & inhibidores , 5'-Nucleotidasa/metabolismo , Humanos , Neutrófilos/efectos de los fármacos , Nucleósido-Trifosfatasa/metabolismo , Proteínas Protozoarias/genética , Proteínas Protozoarias/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Tricomoniasis/parasitología , Trichomonas vaginalis/efectos de los fármacos , Trichomonas vaginalis/crecimiento & desarrollo , Trichomonas vaginalis/metabolismo , Trofozoítos/efectos de los fármacos , Trofozoítos/enzimología , Trofozoítos/crecimiento & desarrollo , Trofozoítos/metabolismo
9.
BMC Infect Dis ; 20(1): 419, 2020 Jun 16.
Artículo en Inglés | MEDLINE | ID: mdl-32546213

RESUMEN

BACKGROUND: Four new variants of Chlamydia trachomatis (nvCTs), detected in several countries, cause false-negative or equivocal results using the Aptima Combo 2 assay (AC2; Hologic). We evaluated the clinical sensitivity and specificity, as well as the analytical inclusivity and exclusivity of the updated AC2 for the detection of CT and Neisseria gonorrhoeae (NG) on the automated Panther system (Hologic). METHODS: We examined 1004 clinical AC2 samples and 225 analytical samples spiked with phenotypically and/or genetically diverse NG and CT strains, and other potentially cross-reacting microbial species. The clinical AC2 samples included CT wild type (WT)-positive (n = 488), all four described AC2 diagnostic-escape nvCTs (n = 170), NG-positive (n = 214), and CT/NG-negative (n = 202) specimens. RESULTS: All nvCT-positive samples (100%) and 486 (99.6%) of the CT WT-positive samples were positive in the updated AC2. All NG-positive, CT/NG-negative, Trichomonas vaginalis (TV)-positive, bacterial vaginosis-positive, and Candida-positive AC2 specimens gave correct results. The clinical sensitivity and specificity of the updated AC2 for CT detection was 99.7 and 100%, respectively, and for NG detection was 100% for both. Examining spiked samples, the analytical inclusivity and exclusivity were 100%, i.e., in clinically relevant concentrations of spiked microbe. CONCLUSIONS: The updated AC2, including two CT targets and one NG target, showed a high sensitivity, specificity, inclusivity and exclusivity for the detection of CT WT, nvCTs, and NG. The updated AC2 on the fully automated Panther system offers a simple, rapid, high-throughput, sensitive, and specific diagnosis of CT and NG, which can easily be combined with detection of Mycoplasma genitalium and TV.


Asunto(s)
Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Análisis de Secuencia de ARN/métodos , Candida/genética , Candidiasis/diagnóstico , Candidiasis/microbiología , Infecciones por Chlamydia/microbiología , Reacciones Cruzadas , Femenino , Gonorrea/diagnóstico , Gonorrea/microbiología , Humanos , Neisseria gonorrhoeae/genética , ARN Bacteriano/genética , ARN Ribosómico 23S/genética , Sensibilidad y Especificidad , Tricomoniasis/diagnóstico , Tricomoniasis/parasitología , Trichomonas vaginalis/genética
10.
BMC Infect Dis ; 20(1): 375, 2020 May 27.
Artículo en Inglés | MEDLINE | ID: mdl-32460721

RESUMEN

BACKGROUND: Sexually transmitted infections (STIs) cause a major public health problem that affect both men and women in developing and developed countries. The aim of the study was to estimate the prevalence of 11 STIs among women who voluntarily participated in the study, while seeking gynecological checkup. The existence of an association between the presence of pathogens and symptoms and various sociodemographic risk factors was assessed. METHODS: A total of 505 vaginal and cervical specimens were collected from women above 18 years of age, with or without symptoms related to gynecological infections. Nucleic acid was extracted and samples were tested by real-time PCR for the following pathogens: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Urealplasma parvum, Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma girerdii, Gardnerella vaginalis, Candida albicans and Human Papillomavirus (HPV). Positive HPV samples underwent genotyping using a microarray system. RESULTS: Of the 505 samples, 312 (62%) were screened positive for at least one pathogen. Of these, 36% were positive for Gardnerella vaginalis, 35% for Ureaplasma parvum, 8% for Candida albicans, 6.7% for HPV, 4.6% for Ureaplasma urealyticum, 3.6% for Mycoplasma hominis, 2% for Trichomonas vaginalis, 0.8% for Chlamydia trachomatis, 0.4% for Mycoplasma girerdii, 0.2% for Mycoplasma genitalium and 0.2% for Neisseria gonorrhoeae. Lack of symptoms was reported in 187 women (37%), among whom 61% were infected. Thirty-four samples were HPV positive, with 17 high risk HPV genotypes (HR-HPV); the highest rates being recorded for types 16 (38%), 18 (21%) and 51 (18%). Out of the 34 HPV positives, 29 participants had HR-HPV. Association with various risk factors were reported. CONCLUSIONS: This is the first study that presents data about the presence of STIs among women in Lebanon and the MENA region by simultaneous detection of 11 pathogens. In the absence of systematic STI surveillance in Lebanon, concurrent screening for HPV and PAP smear is warranted.


Asunto(s)
Enfermedades de Transmisión Sexual/epidemiología , Adulto , Cuello del Útero/microbiología , Cuello del Útero/parasitología , Cuello del Útero/virología , Chlamydia trachomatis/genética , Chlamydia trachomatis/aislamiento & purificación , Estudios Transversales , Femenino , Gardnerella vaginalis/genética , Gardnerella vaginalis/aislamiento & purificación , Humanos , Líbano/epidemiología , Masculino , Epidemiología Molecular , Infecciones por Mycoplasma/epidemiología , Mycoplasma genitalium/genética , Mycoplasma genitalium/aislamiento & purificación , Mycoplasma hominis/genética , Mycoplasma hominis/aislamiento & purificación , Neisseria gonorrhoeae/genética , Neisseria gonorrhoeae/aislamiento & purificación , Papillomaviridae/genética , Papillomaviridae/aislamiento & purificación , Factores de Riesgo , Enfermedades de Transmisión Sexual/microbiología , Enfermedades de Transmisión Sexual/parasitología , Enfermedades de Transmisión Sexual/virología , Trichomonas vaginalis/genética , Trichomonas vaginalis/aislamiento & purificación , Ureaplasma/genética , Ureaplasma/aislamiento & purificación , Vagina/microbiología , Vagina/parasitología , Vagina/virología , Frotis Vaginal , Adulto Joven
11.
Parasitol Res ; 119(6): 1915-1923, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32405804

RESUMEN

Trichomoniasis is the most prevalent curable sexually transmitted infection (STI) worldwide and a risk factor for the acquisition of other STIs and adverse pregnancy outcomes. The objectives of this study were to determine the prevalence of T. vaginalis and related coinfections in women attending a third-level hospital of Madrid (Spain). A retrospective study of 24,173 vaginal exudates from women with suspected vaginitis was conducted between 2013 and 2017. Likewise, among T. vaginalis positive samples, co-occurrence with gonorrhea, chlamydia, syphilis, VIH, Mycoplasma hominis, and Ureaplasma urealyticum was checked. Moreover, seven T. vaginalis isolates from 2017 were randomly collected for endobionts, drug resistance, and microsatellite (MS) instability determinations. The prevalence of T. vaginalis was 0.8% between 2013 and 2017. Less than 20% of patients with trichomoniasis were submitted to a complete screening for other genital pathogens. From that, two patients were coinfected with chlamydia and three with syphilis. Surprisingly, 6.4% of positive samples were diagnosed among pregnant women, showing an alarming increase from 3.2% (2014) to 10% (2017). Among the isolates randomly analyzed, five carried T. vaginalis virus, five harbored mycoplasmas, and one was metronidazole-resistant. The molecular genotyping showed a high variability in the three MS evaluated. To our knowledge, this is the first study in Spain that evaluates the prevalence of trichomoniasis in general and pregnant population and includes biomolecular determinations. These results warn about the increasing prevalence and highlight the importance of including T. vaginalis detection in routine gynecological revisions with special emphasis on childbearing age women and patients with previous STIs.


Asunto(s)
Metronidazol/farmacología , Simbiosis , Centros de Atención Terciaria , Tricomoniasis/epidemiología , Trichomonas vaginalis , Adulto , Coinfección , Resistencia a Medicamentos , Femenino , Gonorrea/complicaciones , Humanos , Persona de Mediana Edad , Mycoplasma hominis/aislamiento & purificación , Embarazo , Complicaciones Parasitarias del Embarazo/epidemiología , Índice de Embarazo , Prevalencia , Estudios Retrospectivos , España/epidemiología , Tricomoniasis/complicaciones , Tricomoniasis/tratamiento farmacológico
12.
BMC Infect Dis ; 20(1): 319, 2020 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-32357839

RESUMEN

BACKGROUND: Trichomoniasis resulting from Trichomonas vaginalis (T. vaginalis) has been considered as a commonly seen disease with the transmission way of sex. At present, the detection methods of T. vaginalis mainly include wet mount microscopy, culture, PCR, immunofluorescence and ELISA. However, all of these detection methods exist shortcomings. METHODS: In this study, a loop-mediated isothermal amplification (LAMP) assay that targeted the species-specific sequence of adhesion protein 65 (AP65) gene had been conducted to detect T. vaginalis. The optimum reaction system and conditions were optimized in this rapid detection method. RESULTS: The results of sensitivity analysis showed that the LAMP assay targeting the AP65 gene was 1000 times more sensitive than the nested PCR targeting the actin gene commonly used for detection of T. vaginalis, and the detecting limitation of the former was 10 trichomonad. Moreover, the amplification of the target gene AP65 by LAMP assay exhibited high specificity and the product was exclusively from T. vaginalis. The detection technique of LAMP did not exhibit cross-reactivity with the common pathogens of Trichinella spiralis, Toxoplasma gondii, Escherichia coli, Candida albicans, Staphylococcus aureus, Haemophilus. CONCLUSIONS: According to the present study, the LAMP assay with the target of AP65 gene, was suitable for the early diagnosis of T. vaginalis infections. Consequently, the LAMP assay was proposed by the current study as a point-of-care examination and an alternative molecular tool which exhibited the potential value in the treatment, control and prevention of trichomoniasis transmission and relevant complication.


Asunto(s)
Moléculas de Adhesión Celular/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa/métodos , Proteínas Protozoarias/genética , Tricomoniasis/diagnóstico , Trichomonas vaginalis/genética , Secuencia de Bases/genética , ADN Protozoario/genética , Diagnóstico Precoz , Femenino , Humanos , Sistemas de Atención de Punto , Sensibilidad y Especificidad , Tricomoniasis/parasitología , Frotis Vaginal
13.
Korean J Parasitol ; 58(2): 135-145, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32418382

RESUMEN

Infections caused by Trichomonas vaginalis in humans are one of the main public health problems caused by sexually transmitted diseases. Objective of this study was to evaluate potential biological activity of the medicinal plant Argemone mexicana (Mexican poppy) on T. vaginalis. Methanolic extracts of the stems and leaves of A. mexicana, and different fractions were prepared with solvents of different polarities. The extracts and functional groups were detected containing sterols, triterpenes, quinones, flavonoids and, alkaloids. Extracts from both the stems and leaves of A. mexicana inhibited the growth of T. vaginalis with half-maximal inhibitory concentration value of 70.6 and 67.2 µg/ml, respectively. In the active fractions, the most abundant compounds were berberine and jatrorrhizine, with presumed antiparasitic activity.


Asunto(s)
Extractos Vegetales/farmacología , Trichomonas vaginalis/efectos de los fármacos , Trichomonas vaginalis/crecimiento & desarrollo , Protocolos de Quimioterapia Combinada Antineoplásica , Vacunas Bacterianas , Ciclofosfamida , Depresión Química , Relación Dosis-Respuesta a Droga , Doxorrubicina , Fluorouracilo , Técnicas In Vitro , Leucovorina , Metanol , Extractos Vegetales/química , Hojas de la Planta/química , Tallos de la Planta/química , Quinonas , Esteroles , Triterpenos
14.
Proc Natl Acad Sci U S A ; 117(23): 13033-13043, 2020 06 09.
Artículo en Inglés | MEDLINE | ID: mdl-32461362

RESUMEN

Trichomonas vaginalis is a common sexually transmitted parasite that colonizes the human urogenital tract causing infections that range from asymptomatic to highly inflammatory. Recent works have highlighted the importance of histone modifications in the regulation of transcription and parasite pathogenesis. However, the nature of DNA methylation in the parasite remains unexplored. Using a combination of immunological techniques and ultrahigh-performance liquid chromatography (UHPLC), we analyzed the abundance of DNA methylation in strains with differential pathogenicity demonstrating that N6-methyladenine (6mA), and not 5-methylcytosine (5mC), is the main DNA methylation mark in T. vaginalis Genome-wide distribution of 6mA reveals that this mark is enriched at intergenic regions, with a preference for certain superfamilies of DNA transposable elements. We show that 6mA in T. vaginalis is associated with silencing when present on genes. Interestingly, bioinformatics analysis revealed the presence of transcriptionally active or repressive intervals flanked by 6mA-enriched regions, and results from chromatin conformation capture (3C) experiments suggest these 6mA flanked regions are in close spatial proximity. These associations were disrupted when parasites were treated with the demethylation activator ascorbic acid. This finding revealed a role for 6mA in modulating three-dimensional (3D) chromatin structure and gene expression in this divergent member of the Excavata.


Asunto(s)
Adenina/metabolismo , Cromatina/química , Metilación de ADN/genética , Trichomonas vaginalis/genética , Ácido Ascórbico/farmacología , Técnicas de Cultivo de Célula , Cromatina/genética , Cromatina/metabolismo , Biología Computacional , Metilación de ADN/efectos de los fármacos , Elementos Transponibles de ADN/genética , Regulación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Conformación Molecular , Análisis de Secuencia de ADN
15.
Obstet Gynecol ; 135(5): 1136-1144, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-32282605

RESUMEN

OBJECTIVE: To identify factors associated with testing for and diagnosis of trichomoniasis in pregnancy and to describe patterns of treatment and tests of reinfection or persistence. METHODS: We conducted a retrospective cohort study of women who delivered from July 2016 to June 2018 at one institution. Testing for Trichomonas vaginalis infection was done by wet mount microscopy or by nucleic acid amplification testing for routine prenatal testing or symptomatic visits. Poisson regression was used to identify factors associated with testing for trichomoniasis and testing positive in pregnancy. Treatment and re-testing patterns also were assessed. RESULTS: Among 3,265 pregnant women, 2,489 (76%) were tested for T vaginalis infection. Of the total sample, 1,808 (55%) were tested by wet mount microscopy, 1,661 (51%) by nucleic acid amplification testing, and 980 (30%) by both modalities. The sensitivity for microscopy compared with nucleic acid amplification testing was 26%, with a specificity of 99%. Factors associated with increased likelihood of being tested included younger age (adjusted risk ratio [aRR] 0.99, 95% CI 0.99-1.00) and bacterial vaginosis (aRR 1.17, 95% CI 1.01-1.37). Prevalence of trichomoniasis was 15% among those tested by any modality (wet mount or nucleic acid amplification testing). Risk factors for trichomoniasis included younger age (aRR 0.97, P<.01), being of black race (aRR 2.62, P<.01), abnormal vaginal discharge (aRR 1.45, P<.01), and chlamydia during the current pregnancy (aRR 1.70, P<.01). Women diagnosed by microscopy had a shorter time to treatment compared with those diagnosed by nucleic acid amplification testing. Most (75%) women with positive infections had a test of reinfection; 29% of these were positive. Bacterial vaginosis was associated with decreased risk of a positive test of reinfection. CONCLUSION: Although testing for and treatment of trichomoniasis during pregnancy is not routinely recommended, the high burden of infection among some pregnant women demonstrates a need to further understand patterns of T vaginalis testing and infection. Opportunities exist for improving timely treatment of trichomoniasis and test of reinfection.


Asunto(s)
Complicaciones Infecciosas del Embarazo/diagnóstico , Diagnóstico Prenatal/métodos , Vaginitis por Trichomonas/diagnóstico , Trichomonas vaginalis , Excreción Vaginal/diagnóstico , Adulto , Femenino , Humanos , Microscopía , Técnicas de Amplificación de Ácido Nucleico , Distribución de Poisson , Embarazo , Complicaciones Infecciosas del Embarazo/epidemiología , Complicaciones Infecciosas del Embarazo/terapia , Prevalencia , Estudios Retrospectivos , Factores de Riesgo , Sensibilidad y Especificidad , Vaginitis por Trichomonas/epidemiología , Vaginitis por Trichomonas/terapia , Excreción Vaginal/epidemiología , Excreción Vaginal/microbiología , Vaginosis Bacteriana/diagnóstico , Vaginosis Bacteriana/epidemiología , Adulto Joven
16.
Wiad Lek ; 73(2): 380-383, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32248179

RESUMEN

OBJECTIVE: The aim was to perform systematic review of genitourinary protozoan invasion and analyze their pathogenicity and the ability to influence the genitourinary infections. PATIENTS AND METHODS: Materials and methods: For systematic review of papers the EMBASE and PubMed databases were searched. We also reviewed our own pilot studies using real-time polymerase chain reaction (PCR) to determine Trichomonas tenax, Pentatrichomonas hominis and Giardia lamblia. CONCLUSION: Conclusions: Trichomonas tenax, Pentatrichomonas hominis, Giardia lamblia can cause genitourinary invasion in addition to Trichomonas vaginalis. Their eradication is obligatory at least for not keeping intact pathogenic microorganisms phagocyted by Trichomonas spp. Defining the protozoan forms is important in preventing of genital infections recurrences and reinfections.


Asunto(s)
Trichomonas vaginalis , Genitales , Giardia , Humanos
17.
Rev. chil. obstet. ginecol. (En línea) ; 85(2): 175-184, abr. 2020. tab
Artículo en Español | LILACS | ID: biblio-1115514

RESUMEN

OBJETIVO: revisar los diferentes métodos de diagnóstico de la tricomoniasis vaginal disponibles hasta el presente. MATERIALES Y MÉTODOS: se revisó la bibliografía latinoamericano e internacional a través de los sitios electrónicos de Pub-Med y Scielo. RESULTADOS: la Tricomonas vaginalis es considera como la enfermedad de transmisión sexual no viral, curable más frecuente y prevalente en el mundo. Se revisan los diferentes de métodos para diagnosticar la presencia de la tricomonas vaginalis en pacientes femeninos con síntomas y signos de la infección producida por el protozoario flagelado. CONCLUSIONES: se revisaron los diferentes métodos de diagnostico de la infección producida por la Tricomonas vaginalis en pacientes femeninas, desde los clásicos hasta los más actuales que emplean alta tecnología.


OBJECTIVE: to review the different diagnostic methods of Trichomonas vaginal available at the present time. MATERIAL AND METHOD: it was reviewed the Latin-American and international bibliography using the Pub-Med and Scielo web sites. RESULTS: Trichomonas vaginalis is considered the most common and prevalent sexual transmitted disease curable and non-viral worldwide. It was reviewed the different methods to diagnose the presence of Trichomonas vaginalis in female patients with symptoms and signs of infection produces by the flagellate protozoa. CONCLUSION: Different methods of diagnosis of the infection produced by Trichomonas vaginalis, since the classics to the most current methods that use high technology, were reviewed.


Asunto(s)
Humanos , Femenino , Vaginitis por Trichomonas/diagnóstico , Trichomonas vaginalis , Frotis Vaginal , Enfermedades de Transmisión Sexual/diagnóstico , Sensibilidad y Especificidad , Técnicas de Amplificación de Ácido Nucleico , Técnicas de Cultivo , Antígenos/análisis
18.
Parasitol Int ; 76: 102086, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32112829

RESUMEN

Trichomonas vaginalis is the protozoan parasite responsible for the most prevalent, non-viral, sexually transmitted disease, which affects millions of people around the world. The main treatment against this disease is metronidazole and some other nitroimidazole derivatives. However, between five and 20% of clinical cases of trichomoniasis are caused by parasites resistant to these drugs. Here we present three compounds that were selected using an innovative strategy, to propose them as possible drugs to combat trichomoniasis, using the glycolytic enzyme triose phosphate isomerase (TvTIM) as the drug target. In the genome of Trichomonas vaginalis there are two genes that encode for two isoforms of TvTIM, known as TvTIM1 and TvTIM2, varying by four out of 254 aminoacid residues. In this study, we used high-throughput virtual screening to search molecules that bind specifically to TvTIM isoforms, in which 34 compounds were selected from a library of nearly 450,000 compounds. The effects of the 34 compounds on the conformation and enzymatic activity of both TvTIM isoforms and their human homolog (HsTIM) were evaluated. We found three compounds that bind specifically, modify the conformation and inhibit TvTIM2 only; although the sequence of both isoforms of TvTIM is almost identical. The selectivity of these compounds towards TvTIM2 is explained by the lower conformational stability of this isoform and that these interactions can inhibit the activity of this enzyme and have an effect against this parasite. These compounds represent promising alternatives for the development of new therapeutic strategies against trichomoniasis.


Asunto(s)
Antiprotozoarios/farmacología , Tricomoniasis/prevención & control , Trichomonas vaginalis/efectos de los fármacos , Triosa-Fosfato Isomerasa/antagonistas & inhibidores , Ensayos Analíticos de Alto Rendimiento , Humanos
19.
PLoS Negl Trop Dis ; 14(3): e0008126, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-32196489

RESUMEN

Trichomonas vaginalis (Tv), a protozoan parasite causing sexually-transmitted disease, has been detected in tissue of prostatitis, benign prostatic hyperplasia (BPH) and prostate cancer (PCa). IL-6, a mediator of chronic inflammation, induces the progression of prostate cancer, and influences the polarization of M2 macrophages, which are the main tumor-associated macrophages. We investigated whether IL-6 produced by human prostate epithelial cells stimulated with Tv induces the M2 polarization of THP-1-derived macrophages, which in turn promotes the progression of PCa. Conditioned medium was prepared from Tv-infected (TCM) and uninfected (CM) prostate epithelial cells (RWPE-1). Thereafter conditioned medium was prepared from macrophages after incubation with CM (M-CM) or TCM (M-TCM). RWPE-1 cells infected with Tv produced IL-6 and chemokines such as CCL2 and CXCL8. When human macrophages were treated with conditioned medium of RWPE-1 cells co-cultured with Tv (TCM), they became polarized to M2-like macrophages as indicated by the production of IL-10 and TGF-ß, and the expression of CD36 and arginase-1, which are M2 macrophage markers. Moreover, proliferation of the M2-like macrophages was also increased by TCM. Blockade of IL-6 signaling with IL-6 receptor antibody and JAK inhibitor (Ruxolitinib) inhibited M2 polarization of THP-1-derived macrophages and proliferation of the macrophages. To assess the effect of crosstalk between macrophages and prostate epithelial cells inflamed by Tv infection on the growth of prostate cancer (PCa) cells, PC3, DU145 and LNCaP cells were treated with conditioned medium from THP-1-derived macrophages stimulated with TCM (M-TCM). Proliferation and migration of the PCa cells were significantly increased by the M-TCM. Our findings suggest that IL-6 produced in response to Tv infection of the prostate has an important effect on the tumor microenvironment by promoting progression of PCa cells following induction of M2 macrophage polarization.


Asunto(s)
Células Epiteliales/metabolismo , Interleucina-6/metabolismo , Macrófagos/efectos de los fármacos , Neoplasias de la Próstata/patología , Prostatitis/complicaciones , Tricomoniasis/complicaciones , Trichomonas vaginalis/inmunología , Animales , Técnicas de Cultivo de Célula , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Medios de Cultivo Condicionados , Progresión de la Enfermedad , Humanos , Masculino , Modelos Teóricos , Células Tumorales Cultivadas
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