Trichomonicidal and parasite membrane damaging activity of bidesmosic saponins from Manilkara rufula

The infection caused by Trichomonas vaginalis is the most common but overlooked nonviral sexually transmitted disease worldwide. Treatment relies on one class of drugs, the 5- nitroimidazoles, but resistance is widespread. New drugs are urgently needed. We reported the effect of crude and purified saponin fractions of Manilkara rufula against Trichomonas vaginalis. The compound responsible for antitrichomonal activity was isolated and identified as an uncommon bidesmosic saponin, Mi-saponin C. This saponin eliminated parasite viability without toxicity against the human vaginal epithelial line (HMVII). In addition, the isolated saponin fraction improved the metronidazole effect against a metronidazole-resistant isolate and dramatically reduced the cytoadherence of T. vaginalis to human cells. Investigation of the mechanism of death showed that the saponin fraction induced the parasite death due to profound membrane damage, inducing a disturbance of intracellular content without nuclear damage. To the best of our knowledge, this is the first report of antitrichomonal activity in the bidesmosic saponins of Manilkara rufula

The Enhanced Activity of a Plant Mixture from the Brazilian Caatinga Biome against Venereal Trichomonads Confirms the Traditional Use

Women living in the semi-arid region of Caatinga in the northeast of Brazil report the use of plant mixtures to treat diseases in the genitourinary tract. Plant extracts were obtained from barks to simulate traditional use. The anti-trichomonads activity as well as the cytotoxic effect of plant extracts were tested. Herein, we confirmed this traditional knowledge by testing plants aqueous extracts against Trichomonas vaginalis and Tritrichomonas foetus, the etiologic agents of human and bovine trichomoniasis. All plant extracts were active individually against at least one trichomonads species except for Prosopis juliflora and Amburana cearensis. Cedrela sp. was the most active against both trichomonads species. Finally, a mixture of plants used in traditional medicine was evaluated for activity. A mixture containing extracts of the plants Ximenia americana, Anadenanthera colubrina var. cebil, Myracrodruon urundeuva, Sideroxylon obtusifolium, and Amburana cearensis was active against the two trichomonads. This finding confirms the traditional practice by women living in the Caatinga region of using a mixture of plants during sitz baths to treat vaginal infections. Altogether, these results highlight the ethnopharmacological use of Cedrela sp. and of the plant mixture for the treatment of venereal diseases by Caatinga residents

Trichomonicidal and parasite membrane damaging activity of bidesmosic saponins from Manilkara rufula

The infection caused by Trichomonas vaginalis is the most common but overlooked nonviral sexually transmitted disease worldwide. Treatment relies on one class of drugs, the 5- nitroimidazoles, but resistance is widespread. New drugs are urgently needed. We reported the effect of crude and purified saponin fractions of Manilkara rufula against Trichomonas vaginalis. The compound responsible for antitrichomonal activity was isolated and identified as an uncommon bidesmosic saponin, Mi-saponin C. This saponin eliminated parasite viability without toxicity against the human vaginal epithelial line (HMVII). In addition, the isolated saponin fraction improved the metronidazole effect against a metronidazole-resistant isolate and dramatically reduced the cytoadherence of T. vaginalis to human cells. Investigation of the mechanism of death showed that the saponin fraction induced the parasite death due to profound membrane damage, inducing a disturbance of intracellular content without nuclear damage. To the best of our knowledge, this is the first report of antitrichomonal activity in the bidesmosic saponins of Manilkara rufula

Iron from haemoglobin and haemin modulates nucleotide hydrolysis in Trichomonas vaginalis

Extracellular ATP may act as a danger signalling molecule, inducing inflammation and immune responses in infection sites. The ectonucleotidases NTPDase and ecto-5’-nucleotidase are enzymes that modulate extracellular nucleotide levels; these enzymes have been previously characterised in Trichomonas vaginalis. Iron plays an important role in the complex trichomonal pathogenesis. Herein, the effects of iron on growth, nucleotide hydrolysis and NTPDase gene expression in T. vaginalis isolates from female and male patients were evaluated. Iron from different sources sustained T. vaginalis growth. Importantly, iron from haemoglobin (HB) and haemin (HM) enhanced NTPDase activity in isolates from female patients and conversely reduced the enzyme activity in isolates from male patients. Iron treatments could not alter the NTPDase transcript levels in T. vaginalis. Furthermore, our results reveal a distinct ATP, ADP and AMP hydrolysis profile between isolates from female and male patients influenced by iron from HB and HM. Our data indicate the participation of NTPDase and ecto-5’-nucleotidase in the establishment of trichomonas infection through ATP degradation and adenosine production influenced by iron

Iron from haemoglobin and haemin modulates nucleotide hydrolysis in Trichomonas vaginalis

Extracellular ATP may act as a danger signalling molecule, inducing inflammation and immune responses in infection sites. The ectonucleotidases NTPDase and ecto-5’-nucleotidase are enzymes that modulate extracellular nucleotide levels; these enzymes have been previously characterised in Trichomonas vaginalis. Iron plays an important role in the complex trichomonal pathogenesis. Herein, the effects of iron on growth, nucleotide hydrolysis and NTPDase gene expression in T. vaginalis isolates from female and male patients were evaluated. Iron from different sources sustained T. vaginalis growth. Importantly, iron from haemoglobin (HB) and haemin (HM) enhanced NTPDase activity in isolates from female patients and conversely reduced the enzyme activity in isolates from male patients. Iron treatments could not alter the NTPDase transcript levels in T. vaginalis. Furthermore, our results reveal a distinct ATP, ADP and AMP hydrolysis profile between isolates from female and male patients influenced by iron from HB and HM. Our data indicate the participation of NTPDase and ecto-5’-nucleotidase in the establishment of trichomonas infection through ATP degradation and adenosine production influenced by iron

Trichomonicidal and parasite membrane damaging activity of bidesmosic saponins from <i>Manilkara rufula</i>

<div><p>The infection caused by <i>Trichomonas vaginalis</i> is the most common but overlooked non-viral sexually transmitted disease worldwide. Treatment relies on one class of drugs, the 5-nitroimidazoles, but resistance is widespread. New drugs are urgently needed. We reported the effect of crude and purified saponin fractions of <i>Manilkara rufula</i> against <i>Trichomonas vaginalis</i>. The compound responsible for antitrichomonal activity was isolated and identified as an uncommon bidesmosic saponin, Mi-saponin C. This saponin eliminated parasite viability without toxicity against the human vaginal epithelial line (HMVII). In addition, the isolated saponin fraction improved the metronidazole effect against a metronidazole-resistant isolate and dramatically reduced the cytoadherence of <i>T</i>. <i>vaginalis</i> to human cells. Investigation of the mechanism of death showed that the saponin fraction induced the parasite death due to profound membrane damage, inducing a disturbance of intracellular content without nuclear damage. To the best of our knowledge, this is the first report of antitrichomonal activity in the bidesmosic saponins of <i>Manilkara rufula</i>.</p></div

Effect of the H100 fraction on <i>T</i>. <i>vaginalis</i> TV-LACM6 isolate adhesion to HMVII and HeLa lineages.

<p>*treatments were statistically different from control. Data are the mean ± SD of at least three different experiments (parasite suspensions) performed in triplicate.</p

TEM of <i>T</i>. <i>vaginalis</i> under H100 fraction treatment.

<p>Control conditions presented one nucleus (N), hydrogenosomes (H) and anterior flagella (AF) (A-B); after 24 h of H100 treatment, trophozoites lost their classical morphology, hydrogenosomes were found in the extracellular medium (EM), and vacuoles (V), and arrows indicate membrane rupture (C-H).</p

<i>Trichomonas vaginalis</i> promotes ROS generation in neutrophils.

<p>Co-incubation of human neutrophils with <i>T</i>. <i>vaginalis</i> treated with H100 and MTZ for 30 min and 10 μM DCF-DA for quantification of ROS production. *<i>P</i> < 0.05 versus neutrophils alone; 5.0 mM H₂O₂ is a positive control.</p