Effect of fungal indoor air pollutant 1-octen-3-ol on levels of reactive oxygen species and nitric oxide as well as dehydrogenases activities in drosophila melanogaster males.

Fungal pollution of indoor environments contributes to several allergic symptoms and represents a public health problem. It is well-established that 1-octen-3-ol, also known as mushroom alcohol, is a fungal volatile organic compound (VOC) commonly found in damp indoor spaces and responsible for the typical musty odor. Previously it was reported that exposure to 1-octen-3-ol induced inflammations and disrupted mitochondrial morphology and bioenergetic rate in Drosophila melanogaster. The aim of this study was to examine the influence of 1-octen-3-ol on dehydrogenase activity, apoptotic biomarkers, levels of nitric oxide (NO) and reactive oxygen species (ROS), as well as antioxidant enzymes activities. D. melanogaster flies were exposed to an atmosphere containing 1-octen-3-ol (2.5 or ∞l/L) for 24 hr. Data demonstrated that 1-octen-3-ol decreased dehydrogenases activity and NO levels but increased ROS levels accompanied by stimulation of glutathione-S-transferase (GST) and superoxide dismutase (SOD) activities without altering caspase 3/7 activation. These findings indicate that adverse mitochondrial activity effects following exposure of D. melanogaster to 1-octen-3-ol, a fungal VOC, may be attributed to oxidant stress. The underlying mechanisms involved in adverse consequences of indoor fungal exposure appear to be related to necrotic but not apoptotic mechanisms. The adverse consequences were sex-dependent with males displaying higher sensitivity to 1-octen-3-ol. Based upon on the fact that the fly genome shares nearly 75% of disease-related genes to human exposure to this fungus may explain the adverse human responses to mold especially for males.

Caatinga plants: Natural and semi-synthetic compounds potentially active against Trichomonas vaginalis.

Trichomonas vaginalis causes trichomoniasis; the most common but overlooked non-viral sexually transmitted disease worldwide. The treatment is based at 5'-nitroimidazoles, however, failure are related to resistance of T. vaginalis to chemotherapy. Caatinga is a uniquely Brazilian region representing a biome with type desert vegetation and plants present diverse biological activity, however, with few studies. The aim of this study was to investigate the activity against T. vaginalis of different plants from Caatinga and identify the compounds responsible by the activity. A bioguided fractionation of Manilkara rufula was performed and four major compounds were identified: caproate of α-amyrin (1b), acetate of ß-amyrin (2a), caproate of ß-amyrin (2b), and acetate of lupeol (3a). In addition, six derivatives of α-amyrin (1), ß-amyrin (2) and lupeol (3) were synthesized and tested against the parasite. Ursolic acid (5) reduced about 98% of parasite viability after 2h of incubation and drastic ultrastructural alterations were observed by scanning electron microscopy. Moreover, 5 presented high cytotoxicity to HMVII and HeLa cell line and low cytotoxicity against Vero line at 50 µM (MIC against the parasite). Metronidazole effect against T. vaginalis resistant isolate was improved when in association with 5.

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.

Natural and synthetic compound anti-Trichomonas vaginalis: an update review.

Trichomonas vaginalis is a flagellate protozoan that causes trichomonosis, a sexually transmitted disease of worldwide importance. However, the infection has long received much less attention than other parasitic and sexually transmitted diseases. This negligence leads to poor diagnosis and underestimated prevalence values, and consequently, it has been associated to increasing acquisition and transmission of HIV, pregnancy outcomes, infertility, pelvic inflammatory disease, and cervical and prostate cancer. In view of increased resistance to drugs belonging to the nitroimidazole class, new treatment alternatives are urgently needed. Natural products provide an immeasurable wealth of active molecules, and a great number of new drugs have been originated from these compounds. In addition, new synthetic products or derivatives from old drugs also provide an alternative to treat trichomonosis. Albeit many studies have been performed with natural products against T. vaginalis, none of them progressed to clinical trials. Overall, inadequate financial investments are made, and no alternative treatment for trichomonosis has been discovered; meanwhile, hundreds of thousands of people will remain infected and suffering the serious consequences of this nonviral STD. Thus, it is highlighted that clinical trials for better understanding the potential in vitro are necessary and urgent in order to furnish a new therapeutic alternative for trichomonosis treatment. The current review attempts to give an overview on the potential of natural and synthetic products as antitrichomonal.

Anti-Trichomonas vaginalis activity from triterpenoid derivatives.

Trichomonas vaginalis is a flagellated parasite that causes trichomonosis, the most common non-viral sexually transmitted disease (STD) in the world. Worryingly, trichomonosis is associated to increased transmission of HIV. Due to high frequency of the infection during pregnancy and the development of metronidazole-resistant isolates, therapeutic alternatives to 5-nitroimidazole are being searched. Triterpenes are natural products presenting several biological activities such as anti-protozoal activity. The aim of this study was to evaluate the in vitro anti-T. vaginalis activity from betulinic and ursolic acids, as well as semisynthetic derivatives obtained. Compounds obtained from betulinic acid presented better activity than those from ursolic acid. Piperazine derivatived from betulinic acid presented minimum inhibitory concentration (MIC) value of 91.2 µM, and the kinetic growth curve performed with parasites treated with this most active compound revealed complete inhibition of trophozoite proliferation at 2 h of incubation and total abolition of trophozoite growth in 24 h, revealing that the piperazine derivative is an efficient trichomonacidal molecule. The same compound promoted total erythrocyte lysis and lactate dehydrogenase (LDH) liberation of 83 and 100% (at 45.6 and 91.2 µM, respectively), indicating parasite membrane damage. The piperazine derivative demonstrated cytotoxic effect against the HMVII and HeLa cell lineages at the MIC value. This is the first report of semisynthetic triterpenoid derivatives with anti-T. vaginalis activity, revealing the high potential of these compounds as trichomonacidal agents.

Sleep disturbance induces a modulation of clock gene expression and alters metabolism regulation in drosophila.

Sleep disorders are catching attention worldwide as they can induce dyshomeostasis and health issues in all animals, including humans. Circadian rhythms are biological 24-hour cycles that influence physiology and behavior in all living organisms. Sleep is a crucial resting state for survival and is under the control of circadian rhythms. Studies have shown the influence of sleep on various pathological conditions, including metabolic diseases; however, the biological mechanisms involving the circadian clock, sleep, and metabolism regulation are not well understood. In previous work, we standardized a sleep disturbance protocol and, observed that short-time sleep deprivation and sleep-pattern alteration induce homeostatic sleep regulation, locomotor deficits, and increase oxidative stress. Now, we investigated the relationship between these alterations with the circadian clock and energetic metabolism. In this study, we evaluated the expression of the circadian clock and drosophila insulin-like peptides (DILPs) genes and metabolic markers glucose, triglycerides, and glycogen in fruit flies subjected to short-term sleep disruption protocols. The sleep disturbance altered the expression of clock genes and DILPs genes expression, and modulated glucose, triglycerides, and glycogen levels. Moreover, we demonstrated changes in mTor/dFoxo genes, AKT phosphorylation, and dopamine levels in nocturnal light-exposed flies. Thus, our results suggest a connection between clock genes and metabolism disruption as a consequence of sleep disruption, demonstrating the importance of sleep quality in health maintenance.

Screening and bioguided fractionation of Amaryllidaceae species with anti-Trichomonas vaginalis activity.

The Amaryllidaceae family is known by its ornamental and medicinal value and has attracted considerable attention due to the content of alkaloids of its species, which showed interesting biological properties. The present study evaluated the anti-Trichomonas vaginalis activity of eighteen extracts (12.5 to 0.19 mg/mL) and six isolated alkaloids (125 to 1.9 µg/mL) from Amaryllidaceae species. The alkaloids diminished the trophozoites viability (from 15 to 40 %). The extracts from Hippeastrum breviflorum demonstrated the highest anti-T. vaginalis activity (viability was 60 % reduced), and a bioguided study was conducted. Six fractions with antiprotozoal activity had lycorine and lycosinine as major components suggesting a synergistic effect, taking into account the higher anti-T. vaginalis activity of extracts when compared to isolated alkaloids. Our results point out the antiprotozoal potential of the Amaryllidaceae species against T. vaginalis. This parasite causes trichomonosis, the most prevalent nonviral sexually transmitted diseases (STD) worldwide and a public health problem that requires new therapeutic alternatives as well bioactive natural products.

Candimine-induced cell death of the amitochondriate parasite Trichomonas vaginalis.

Candimine (1), an alkaloid from the bulbs of Hippeastrum morelianum, was found to be cytotoxic for the amitochondriate parasite Trichomonas vaginalis. Candimine (1) induced cell death with an unprecedented group of effects that failed to fulfill the criteria for apoptosis and apoptosis-like death already reported in trichomonads. Arrest of the parasite cell cycle, and morphologic and ultrastructural alterations, including marked cytoplasmic vacuolization, were induced by 1. The present findings suggest some similarities to paraptotic cell death, described for multicellular organisms. This study contributes to both a better understanding of the biological effects of 1 and T. vaginalis cell biology.

Remarkable anti-trichomonas vaginalis activity of plants traditionally used by the Mbyá-Guarani indigenous group in Brazil.

Trichomonas vaginalis, a flagellate protozoan, is the causative agent of trichomonosis, the most common nonviral sexually transmitted disease worldwide. Taking into account the increased prevalence of metronidazole-resistant isolates, alternative drugs are essential for the successful treatment. Natural products are the source of most new drugs, and popular wisdom about the use of medicinal plants is a powerful tool in this search. In this study, the activity of 10 medicinal plants extensively used in daily life by Mbyá-Guarani indigenous group was evaluated against seven different T. vaginalis isolates. Among the aqueous extracts tested, Verbena sp. (Guachu ka'a in Mbyá-Guarani language) and Campomanesia xanthocarpa (Guavira in Mbyá-Guarani language) showed the highest activity against T. vaginalis with MIC value of 4.0 mg/mL reaching 100% of efficacy against the parasite. The kinetic growth assays showed that the extracts promoted complete growth abolishment after 4 h of incubation. In addition, the extracts tested did not promote a significant hemolytic activity against human erythrocytes. Our results show for the first time the potential activity of Verbena sp. and C. xanthocarpa against T. vaginalis. In addition, this study demonstrates that indigenous knowledge is an important source of new prototype antiprotozoal agents.

Anti-Trichomonas vaginalis activity of Hypericum polyanthemum extract obtained by supercritical fluid extraction and isolated compounds.

The anti-Trichomonas vaginalis activity of Hypericum polyanthemum extract obtained by supercritical fluid extraction (50°C, 150bar) and the chemical compounds isolated and purified from this extract (benzopyrans HP1, HP2, HP3, and phloroglucinol derivative uliginosin B) were assessed. All samples had anti-T. vaginalis activity; however, HP1 demonstrated the best selectivity against this protozoan (metronidazole-resistant and susceptible isolates), with no cytotoxicity on mammalian cells (selectivity index of 73.97). Moreover, HP1 had activity against a metronidazole-resistant isolate (52% of viable trophozoites), and this effect was higher when tested with a low concentration of metronidazole (23% of viable trophozoites). Experiments demonstrated that all isolated compounds caused damage to the parasites' membrane (>90% of LDH release) and do not present a notable hemolytic effect, although HP2 and uliginosin B exhibited cytotoxicity against mammalian cells. Therefore, the analyzed molecules are promising prototypes for new antiprotozoal drugs, especially HP1, which seems to improve metronidazole's effect on a resistant T. vaginalis isolate.

Lycorine induces cell death in the amitochondriate parasite, Trichomonas vaginalis, via an alternative non-apoptotic death pathway.

In this study, the mechanism of action of the pro-apoptotic alkaloid lycorine on an amitochondriate cell, the parasite Trichomonas vaginalis, was investigated. The cytotoxicity of lycorine against T. vaginalis was studied from 2.5 to 1000µM and several important ultrastructural alterations were observed by electron microscopy. Lycorine arrested the T. vaginalis cell cycle, although no hallmarks of apoptosis, such as apoptotic bodies, were observed. Consequently, the underlying mechanism of action fails to completely fulfill the criteria for apoptosis. However, some similarities to paraptotic cell death were observed.

Kinetic characterization and gene expression of adenosine deaminase in intact trophozoites of Trichomonas vaginalis.

Trichomonas vaginalis is a parasite that resides in the human urogenital tract and causes trichomonosis, the most prevalent nonviral sexually transmitted disease. Nucleoside triphosphate diphosphohydrolase (NTPDase), which hydrolyzes extracellular di- and triphosphate nucleotides, and ecto-5'-nucleotidase, which hydrolyzes AMP, have been characterized in T. vaginalis. The aim of this study was to characterize the adenosine deaminase (ADA) activity in intact trophozoites of T. vaginalis. A strong inhibition in adenosine deamination was observed in the presence of calcium and magnesium, which was prevented by EDTA. The apparent K(M) value for adenosine was 1.13 ± 0.07mM. The calculated V(max) was 2.61 ± 0.054 nmol NH(3) min(-1) mg(-1) protein. Adenosine deamination was inhibited in the presence of erythro-9-(2-hydroxy-3-nonyl)adenine. Semi-quantitative reverse transcriptase-PCR experiments were performed and both ADA-related genes ada(125) and ada(231) mRNA were expressed, although ada(231) in higher quantity when compared with the ada(125) : α-tubulin ratio. Furthermore, a phylogenetic analysis showed that the T. vaginalis sequences formed a clade with Entamoeba histolytica and Dictyostelium discoideum sequences, and it strongly suggests homologous functions in the T. vaginalis genome. The presence of ADA activity in T. vaginalis may be important to modulate the adenosine/inosine levels during infection and, consequently, to maintain the anti-inflammatory properties through different nucleoside-signalling mechanisms.

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.

Mecanismos específicos de patogenicidadede protozoários de mucosa: Entamoeba histolytica, Giardia lamblia e Trichomonas vaginali / Specific pathogenic mechanisms of mucosal protozoans: Entamoeba histolytica, Giardia lamblia and Trichomonas vaginalis

Entamoeba histolytica e Giardia lamblia são protozoários que podem parasitar a mucosa intestinal, causando principalmente diarreia. Trichomonas vaginalis coloniza a mucosa vaginal causando tricomonose, a doença sexualmente transmissível não viral mais comum no mundo. Embora coletivamente estes parasitos infectem mais de um bilhão de pessoas a cada ano, seus mecanismos de patogenicidade ainda não estão totalmente esclarecidos. Assim, esta revisão reúne os principais mecanismos envolvidos na patogenicidade destes protozoários, bem como os fatores do microambiente que podem interferir no sucesso da colonização. A patogênese da E. histolytica envolve adesão, lise, fagocitose de células epiteliais e bactérias, invasão tecidual por ação de enzimas e evasão da resposta imune do hospedeiro. A lectina Gal/GalNAc, os amebaporos e as cisteína proteases são as principais moléculas envolvidas nesses processos. O estabelecimento da giardiose depende de diversos mecanismos patogênicos e de virulência desenvolvidos pela G. lamblia, tais como as moléculas envolvidas na adesão, encistamento e variação antigênica. Para o sucesso da colonização da mucosa vaginal, o T. vaginalis expressa moléculas como as adesinas de superfície, lipofosfoglicanos e galectina, envolvidas na adesão às células epiteliais vaginais e alteração da expressão gênica, tanto do parasito como do hospedeiro.

Entamoeba histolytica and Giardia lamblia are protozoans that may parasitize the intestinal mucosa, mainly causing diarrhea. Trichomonas vaginalis colonizes the vaginal mucosa causing trichomonosis, the most common non-viral sexually transmitted disease in the world. Although collectively these parasites infect over a billion people each year, their pathogenic mechanisms have not been completely understood so far. Hence, this review of the literature demonstrates the main mechanisms involved in the pathogenicity of these protozoans, as well as the microenvironmental factors that can interfere with successful colonization. The pathogenesis of E. histolytica involves adhesion, lysis, phagocytosis of epithelial cells and bacteria, tissue invasion by enzymatic action, and evasion of host immune response. Lectin Gal/GalNac, amoebapores, and cysteine proteases are the main molecules involved in these processes. The establishment of giardiosis depends on several pathogenic mechanisms and virulence developed by G. lamblia, such as molecules involved in adhesion, encystation and antigenic variation. For successful colonization of vaginal mucosa, T. vaginalis express molecules like adhesins on the surface and galectin and lipophosphoglycan, involved in the adherence to vaginal epithelial cells and altered gene expression of both the parasite and the host.