Optimization of Coumarins Extraction from Pterocaulon balansae by Box-Behnken Design and Anti-Trichomonas vaginalis Activity

Planta Medica ◽  
2021 ◽  
Author(s):  
Nathalya Tesch Brazil ◽  
Bruna Medeiros-Neves ◽  
Flávia Nathiely Silveira Fachel ◽  
Vanessa Pittol ◽  
Roselena Silvestri Schuh ◽  
...  

Abstract Trichomonas vaginalis causes trichomoniasis, a nonviral sexually transmitted infection with a high prevalence worldwide. Oral metronidazole is the drug of choice for the treatment of this disease, although high levels of T. vaginalis resistance to this agent are well documented in the literature. This study describes the anti-T. vaginalis activity of an optimized coumarin-rich extract from Pterocaulon balansae. Optimization was performed to maximize extraction of total coumarins by means of a 3-level Box-Behnken design, evaluating the effect of three factors: extraction time, plant : solvent ratio, and ethanol concentration. Optimum conditions were found to be 5 h extraction time and a plant : solvent ratio of 1% (w/v) and 60% (v/v) ethanol, which resulted in approximately 30 mg of total coumarins/g of dry plant. The coumarin-enriched extract exhibited a minimum inhibitory concentration of 30 µg/mL and an IC50 of 3.2 µg/mL against T. vaginalis, a low cytotoxicity, and a high selectivity index (18 for vaginal epithelial cells and 16 for erythrocytes). The coumarins permeation/retention profile through porcine vaginal mucosa was evaluated in Franz-type diffusion cells. After 8 h of kinetics, coumarins were detected in the tissue (4.93 µg/g) without detecting them in the receptor compartment. A significant increase of coumarins in the mucosa layers (8.18 µg/g) and receptor compartment (0.26 µg/g) was detected when a T. vaginalis suspension (2 × 105 trophozoites/mL) was previously added onto the mucosa. No alterations were visualized in the stratified squamous non-keratinized epithelium of the porcine vaginal mucosa after contact with the extract. Overall, these results suggest that the P. balansae coumarin-rich extract may have potential as a treatment for trichomoniasis.

mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Jully Pinheiro ◽  
Jacob Biboy ◽  
Waldemar Vollmer ◽  
Robert P. Hirt ◽  
Jeremy R. Keown ◽  
...  

ABSTRACTThe human eukaryotic pathogenTrichomonas vaginaliscauses trichomoniasis, a prevalent sexually transmitted infection. This extracellular protozoan is intimately associated with the human vaginal mucosa and microbiota, but key aspects of the complex interactions between the parasite and the vaginal bacteria remain elusive. We report thatT. vaginalishas acquired, by lateral gene transfer from bacteria, genes encoding peptidoglycan hydrolases of the NlpC/P60 family. Two of theT. vaginalisenzymes were active against bacterial peptidoglycan, retaining the active-site fold and specificity asdl-endopeptidases. The endogenous NlpC/P60 genes are transcriptionally upregulated inT. vaginalisin the presence of bacteria. The overexpression of an exogenous copy enables the parasite to outcompete bacteria from mixed cultures, consistent with the biochemical activity of the enzyme. Our study results highlight the relevance of the interactions of this eukaryotic pathogen with bacteria, a poorly understood aspect of the biology of this important human parasite.IMPORTANCETrichomonas vaginalisis a parasitic protozoan of the human urogenital tract that causes trichomoniasis, a very common sexually transmitted disease. Despite residing extracellularly and in close association with the vaginal bacteria (i.e., the microbiota), very little is known about the nature of the parasite-bacterium interactions. Our study showed that this parasite had acquired genes from bacteria which retained their original function. They produce active enzymes capable of degrading peptidoglycan, a unique polymer of the bacterial cell envelope, helping the parasite to outcompete bacteria in mixed cultures. This study was the first to show that a laterally acquired group of genes enables a eukaryotic mucosal pathogen to control bacterial population. We highlight the importance of understanding the interactions between pathogens and microbiota, as the outcomes of these interactions are increasingly understood to have important implications on health and disease.


2018 ◽  
Author(s):  
Jully Pinheiro ◽  
Jacob Biboy ◽  
Waldemar Vollmer ◽  
Robert P. Hirt ◽  
Jeremy R. Keown ◽  
...  

AbstractTrichomonas vaginalisis a human eukaryotic pathogen and the causative agent of trichomoniasis, the most prevalent non-viral sexually transmitted infection worldwide. This extracellular protozoan parasite is intimately associated with the human vaginal mucosa and microbiota but key aspects of the complex interactions between the parasite and the vaginal bacteria remain elusive. We report thatT. vaginalishas acquired, by lateral gene transfer from bacteria, genes encoding peptidoglycan hydrolases of the NlpC/P60 family. Two of theT. vaginalisenzymes were active against bacterial peptidoglycan, retaining the active site fold and specificity as DL-endopeptidases. The endogenous NlpC/P60 genes are transcriptionally up regulated inT. vaginaliswhen in the presence of bacteria. The over-expression of an exogenous copy produces a remarkable phenotype where the parasite is capable of competing out bacteria from mixed cultures, consistent with the biochemical activity of the enzymein vitro. Our study highlights the relevance of the interactions of this eukaryotic pathogen with bacteria, a poorly understood aspect on the biology of this important human parasite.Author summaryTrichomonas vaginalisis a protozoan parasite that causes a very common sexually transmitted disease known as trichomoniasis. This extracellular parasite resides in the vagina where it is in close association with the mucosa and the local microbiota. Very little is known about the nature of the parasite-bacteria interactions. Here, we report that this parasite had acquired genes from bacteria which retained their original function producing active enzymes capable of degrading peptidoglycan, a polymer that is chemically unique to the cell envelope of bacteria. Our results indicate that these enzymes help the parasite compete out bacteria in mixed cultures. These observations suggest that these enzymes may be critical for the parasite to establish infection in the vagina, a body site that is densely colonised with bacteria. Our study further highlights the importance of understanding the interactions between pathogens and microbiota, as the outcomes of these interactions are increasingly understood to have important implications on health and disease.


Parasitology ◽  
2018 ◽  
Vol 146 (9) ◽  
pp. 1179-1183 ◽  
Author(s):  
Graziela Vargas Rigo ◽  
Brenda Petro-Silveira ◽  
Michael Devereux ◽  
Malachy McCann ◽  
André Luis Souza dos Santos ◽  
...  

AbstractTrichomonas 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.


Author(s):  
Shiren Ali Al Hamzawi

Estimates of Trichomonas vaginalis prevalence in pregnant women are variable with few studies in Iraq.T. vaginalis is a worldwide prevalent sexually transmitted infection,but fortunately,it is very treatable. Researchers believed that pregnancy is one of the effective factors for T. vaginalis infection in women.A cross-sectional study performed in Obstetrics and Gynecology Department at Maternity and Children Teaching Hospital in Al-Diwaniya city on two hundred female pregnant patients between the ages of 16-45 years. These females had no intercourse for 2–3 days,not using drugs (antibiotics,antiprotozoal or steroids) for the last 15 days. Vaginal discharges of any type with or without itching,burning sensation or both were their main complaints. Vaginal swabs were taken from all participating patients for direct wet mount microscopy and culture for the detection of Trichomonas vaginalis infection. The study showed that twelve out of two hundred examined pregnant women (6%) presented with T. vaginalis infection. The infection was more in those with mothers’ age (26-35) years,housewives,low education,higher parity,and of rural residents. Other maternal variables were not significantly associated with T. vaginalis infection. The study showed a prevalence of (6%) of T. vaginalis infection in pregnant female attendees. Infection was more in those with mothers ’age (26-35) years,housewives,low educational level,higher parity,and living in rural areas.


2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Martin Obermeier ◽  
Monia Pacenti ◽  
Robert Ehret ◽  
Francesco Onelia ◽  
Rory Gunson ◽  
...  

AbstractObjectivesAutomated molecular analyzers have accelerated diagnosis, allowing earlier intervention and better patient follow-up. A recently developed completely automated molecular analyzer, Alinity™ m (Abbott), offers consolidated, continuous, and random-access testing that may improve molecular laboratory workflow.MethodsAn international, multicenter study compared laboratory workflow metrics across various routine analyzers and Alinity m utilizing assays for human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV), hepatitis B virus (HBV), high-risk human papillomavirus (HR HPV), and sexually transmitted infection (STI) (Chlamydia trachomatis [CT]/Neisseria gonorrhoeae [NG]/Trichomonas vaginalis [TV]/Mycoplasma genitalium [MG]). Three turnaround times (TATs) were assessed: total TAT (sample arrival to result), sample onboard TAT (sample loading and test starting to result), and processing TAT (sample aspiration to result).ResultsTotal TAT was reduced from days with routine analyzers to hours with Alinity m, independent of requested assays. Sample onboard TATs for standard workflow using routine analyzers ranged from 7 to 32.5 h compared to 2.75–6 h for Alinity m. The mean sample onboard TAT for STAT samples on Alinity m was 2.36 h (±0.19 h). Processing TATs for Alinity m were independent of the combination of assays, with 100% of results reported within 117 min.ConclusionsThe consolidated, continuous, random-access workflow of Alinity m reduces TATs across various assays and is expected to improve both laboratory operational efficiency and patient care.


Author(s):  
Fatemeh Rahmani ◽  
Yahya Ehteshaminia ◽  
Hamid Mohammadi ◽  
Seif Ali Mahdavi

Introduction: Trichomoniasis is the most common non-viral sexually transmitted infection in the world, caused by the protozoan parasite Trichomonas vaginalis, which infects the urogenital tract of men and women. Approximately, 250 million new cases of Trichomonas vaginalis Infection are reported worldwide each year. Trichomoniasis is also considered an important HIV co-infection. The infection is often asymptomatic but can be accompanied by symptoms such as severe inflammation, itching and irritation, foamy discharge, and malodorous smell mucus, but the signs and symptoms of the disease are not sufficient for specific diagnosis. Material and Methods: In this study, the websites of PubMed, Google Scholar, SID, and Margiran were searched and related articles were reviewed. Results: Only screening and the use of highly sensitive and specific diagnostic methods can identify asymptomatic individuals. Today, the most common way to diagnose the infection is to use wet slide, Pap smear and culture methods that do not have high sensitivity and specificity. Also, due to the increase in infection and its complications, finding an efficient, rapid, and easy test to detect the parasite and differentiate Trichomoniasis vaginitis from other sexually transmitted diseases is considered important and necessary. Conclusion: Nowadays, there are several diagnostic methods that differentiate trichomoniasis infection from other sexually transmitted infections with high accuracy and sensitivity. Of course, existing diagnostic methods mostly use women's urine and vaginal samples for diagnosis, and methods that specifically diagnose the infection in men are more limited.


2020 ◽  
pp. 742-748
Author(s):  
Nada H. Bedair ◽  
Hayder Z. Ali

Trichomonas vaginalis is an eukaryotic parasite that causes the most common non-viral sexually transmitted infection, trichomoniasis. This disease is responsible for many serious health problems such as preterm birth. More than half of the infected women do not develop symptoms, which makes it difficult to diagnose thedisease. In this study, a specific indirect ELISA method was developed to detect anti-Trichomonas vaginalis IgM and IgG immunoglobulins in the sera of infected females. The aim of this study was to investigate the sensitivity of a simple ELISA procedure in comparison to the classical urine examination and vaginal wet mount preparation for the diagnosis of T. vaginalis. The sensitivity of the indirect ELISA was compared with the classical vaginal discharge swab and urine microscopic examination, and the results showed sensitivities of 65.5% and 57.2%, respectively. Furthermore, the infection was measurable as acute or chronic with the refined test.


2020 ◽  
Vol 31 (7) ◽  
pp. 627-636 ◽  
Author(s):  
Pooja Chitneni ◽  
Mags Beksinska ◽  
Janan J Dietrich ◽  
Manjeetha Jaggernath ◽  
Kalysha Closson ◽  
...  

Partner notification and treatment are essential components of sexually transmitted infection (STI) management, but little is known about such practices among adolescents and young adults. Using data from a prospective cohort study (AYAZAZI) of youth aged 16–24 years in Durban, South Africa, we assessed the STI care cascade across participant diagnosis, STI treatment, partner notification, and partner treatment; index recurrent STI and associated factors; and reasons for not notifying partner of STI. Participants completed laboratory-based STI screening ( Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis) at enrollment and at 12 months. Of the 37/216 participants with STI (17%), 27/37 (73%) were women and 10/37 (27%) were men. Median age was 19 years (IQR: 18–20). Of the participants with STI, 23/37 (62%) completed a Treatment and Partner Tracing Survey within 6 months of diagnosis. All survey participants reported completing STI treatment (100%), 17/23 (74%) notified a partner, and 6/23 (35%) reported partner treatment. Overall, 4/23 (11%) participants had 12-month recurrent C. trachomatis infection, with no association with partner notification or treatment. Stigma and lack of STI knowledge were reasons for not notifying partner of STI. STI partner notification and treatment is a challenge among youth. Novel strategies are needed to overcome barriers along the STI care cascade.


2007 ◽  
Vol 76 (2) ◽  
pp. 523-531 ◽  
Author(s):  
V. Mundodi ◽  
A. S. Kucknoor ◽  
J. F. Alderete

ABSTRACT Trichomonas vaginalis is a protist that causes the most common human sexually transmitted infection. A T. vaginalis cDNA expression library was screened with pooled sera from patients with trichomoniasis. A highly reactive cDNA clone of 1,428 bp encoded a trichomonad protein of 472 amino acids with sequence identity to α-enolase (tv-eno1). The sequence alignment confirmed the highly conserved nature of the enzyme with 65% to 84% identity among organisms. The expression of tv-eno1 was up-regulated by contact of parasites with vaginal epithelial cells, and this is the first report demonstrating up-regulation by cytoadherence of a plasminogen-binding α-enolase in T. vaginalis. Immunofluorescence with monoclonal antibody of nonpermeabilized trichomonads showed tv-ENO1 on the surface. The recombinant tv-ENO1 was expressed in Escherichia coli as a glutathione S-transferase (GST)::tv-ENO1 fusion protein, which was cleaved using thrombin to obtain affinity-purified recombinant tv-ENO1 protein (tv-rENO1) detectable in immunoblots by sera of patients. Immobilized tv-rENO1 bound human plasminogen in a dose-dependent manner, and plasminogen binding by tv-rENO1 was confirmed in a ligand blot assay. The plasminogen-specific inhibitor ε-aminocaproic acid blocked the tv-rENO1-plasminogen association, indicating that lysines play a role in binding to tv-rENO1. Further, both parasites and tv-rENO1 activate plasminogen to plasmin that is mediated by tissue plasminogen activator. These data indicate that as with other bacterial pathogens, tv-ENO1 is an anchorless, surface-associated glycolytic enzyme of T. vaginalis.


1996 ◽  
Vol 40 (5) ◽  
pp. 1121-1125 ◽  
Author(s):  
E M Narcisi ◽  
W E Secor

Trichomonas vaginalis is a common sexually transmitted protozoan parasite. Although often considered simply a nuisance infection, T. vaginalis has been implicated in premature rupture of placental membranes and increases in the risk of acquiring human immunodeficiency virus. Metronidazole, a 5-nitroimidazole, is currently the drug of choice to treat T. vaginalis infection. Because some patients have severe reactions to metronidazole and others are infected with metronidazole-resistant T. vaginalis, we were prompted to investigate alternative therapies. Tinidazole, another 5-nitroimidazole used in other countries to treat T. vaginalis infections, and furazolidone, a nitrofuran presently used to treat giardiasis and infections with some anaerobic enteric bacteria, were investigated for effectiveness against 9 metronidazole-susceptible and 12 metronidazole-resistant T. vaginalis patient isolates. The in vitro aerobic and anaerobic minimum lethal concentrations (MLC) and the time for drug efficacy were determined. Tinidazole killed the metronidazole-susceptible isolates at a low MLC but was effective against only 4 of the 12 metronidazole-resistant isolates. In contrast, furazolidone was effective at a low MLC for all isolates. When tinidazole was effective, it required > 6 h to kill trichomonads. However, furazolidone killed both metronidazole-susceptible and resistant trichomonads within 2 to 3 h of exposure. These data suggest that furazolidone may be a good candidate for treating metronidazole-resistant trichomoniasis and that further investigation of this drug is warranted.


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