Cultivation

Linstead, David

doi:10.1007/978-1-4612-3224-7_7

Cited by 11 publications

(5 citation statements)

References 35 publications

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“…For trichomonad culture in vitro, the serum added to the growth medium is extremely important for trophozoite growth maintenance. The main role of the serum is to supply lipids, in particular fatty acids and cholesterol, in a form that can be assimilated by the organism without toxicity (Linstead 1990). However, besides these compounds, serum provides additional molecules that are essential for the parasites, such as precursors of the purine and pyrimidine nucleotides (Wang et al 1984).…”

Section: Discussionmentioning

confidence: 99%

“…The parasites have been shown to be phagocytic and the contribution of phagocytosis to nutrient acquisition is advantageous for the parasite, as the vaginal cavity can be a harsh environment with scarce nutrients (Pereira-Neves & Benchimol 2007). In vitro, parasite culture medium is typically supplemented with bovine, horse or human serum to provide an important nutritional source for the trophozoites (Linstead 1990). In mammalian cells, it has been demonstrated that serum deprivation alters cellular growth and promotes an increase in the incorporation of adenosine into adenine nucleotides (Rapaport & Zamecnik 1978).…”

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confidence: 99%

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Analysis of the NTPDase and ecto-5'-nucleotidase profiles in serum-limited Trichomonas vaginalis

Frasson

Charão

Souza

et al. 2012

Mem. Inst. Oswaldo Cruz

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Trichomonas vaginalis is a parasite of the human urogenital tract that causes trichomonosis, the most prevalent non-viral sexually transmitted disease. Ectonucleoside triphosphate diphosphohydrolase (NTPDase) family members, which hydrolyse extracellular ATP and ADP and ecto-5'-nucleotidase, which hydrolyses AMP, have been characterised in T. vaginalis. For trichomonad culture, the growth medium is supplemented with 10% serum, which is an important source of nutrients, such as adenosine. Here, we investigated the ATP metabolism of T. vaginalis trophozoites from long-term cultures and clinical isolates under limited bovine serum conditions (1% serum). The specific enzymatic activities were expressed as nmol inorganic phosphate (Pi) released/min/mg protein, the gene expression patterns were determined by reverse transcriptase-polymerase chain reaction, the extracellular adenine nucleotide hydrolysis was analysed by high performance liquid chromatography and the cell cycle analysis was assessed by flow cytometry. Serum limitation led to the profound activation of NTPDase and ecto-5'-nucleotidase activities. Furthermore, the levels of NTPDase A and B transcripts increased and extracellular ATP metabolism was activated, which led to enhanced ATP hydrolysis and the formation of ADP and AMP. Moreover, the cell cycle was arrested at the G0/G1 stage, which suggested adenosine uptake. Our data suggest that under conditions of serum limitation, NTPDase and ecto-5'-nucleotidase play a role in providing the adenosine required for T. vaginalis growth and that this process contributes to the establishment of parasitism.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Analysis of the NTPDase and ecto-5'-nucleotidase profiles in serum-limited Trichomonas vaginalis

Frasson

Charão

Souza

et al. 2012

Mem. Inst. Oswaldo Cruz

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“…In vitro pharmaceutical activity of the heterocyclic ingredient Preparation of culture media. For culturing E. histolytica, Boeck and Dr. Bohuslav's Locke-Egg-Serum (LES) medium were used [23]. First, the Locke solution was prepared, filtrated, and autoclaved.…”

Section: In Vivo Pharmaceutical Activitymentioning

confidence: 99%

Pharmaceutical activity of a synthetic heterocyclic (C15H12N5OCl) compound on Entamoeba histolytica and Giardia lamblia

Obaid

Saleh

Boundenga

2023

Russian Journal of Infection and Immunity

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Abstract: Background: Intestinal parasites are among the most important infectious agents with an impact on human health. Indeed, in the lack of an available treatment option, these parasites could constitute a real health problem in the population. Aim: In the present study, we investigated the effect of a synthetic heterocyclic compound on two intestinal parasites (Entamoeda histolytica and Gardia lamblia). Methods: The parasite isolates tested were collected from outpatients at the General Pediatric Hospital in Kirkuk, Iraq, between September 2019 and January 2020. Thus, we studied the in vivo and in vitro pharmaceutical activity of the ingredient on both parasites. The toxicological effects of the substance on some blood parameters and liver and kidney function tests were also studied. Results: After five days of treatment, the drug's in vivo action on G. lamblia resulted in an inhibition rate of 88.2 percent at a dose of 1 mg/kg. On the other hand, we observed that the influence of this synthetic substance on cultured E. histolytica was very close to the metronidazole effect. The maximum result was at a concentration of 1 g/ml and was obtained after 72 hours of incubation with an inhibition rate of 89.4%. The substance did not affect the blood parameters or the studied liver and kidney functions. Conclusion: It can be concluded that this substance is highly effective against both E. histolytica and G. lamblia, and that it has no toxic effects on the studied parameters. Therefore, it could be a promising pharmacophore for intestinal protozoan parasites including E. histolytica and G. lamblia and an alternative or competitor to the current medications available.

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“…In vivo, these nutrients are acquired from vaginal microorganisms and host cells ( Petrin et al 1998 ). In vitro, culture media are typically supplemented with bovine or horse serum, which provides these nutrients ( Linstead 1989 ). Adenosine is the primary precursor of all purine nucleotides in T. vaginalis, but because it lacks de novo purine and pyrimidine synthesis, it depends on the salvage pathway for their acquisition ( Munagala & Wang 2003 ).…”

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confidence: 99%

Modulatory effect of iron chelators on adenosine deaminase activity and gene expression in Trichomonas vaginalis

Primon-Barros

Rigo

Frasson

et al. 2015

Mem. Inst. Oswaldo Cruz

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Trichomonas vaginalis is a flagellate protozoan that parasitises the urogenital human tract and causes trichomoniasis. During the infection, the acquisition of nutrients, such as iron and purine and pyrimidine nucleosides, is essential for the survival of the parasite. The enzymes for purinergic signalling, including adenosine deaminase (ADA), which degrades adenosine to inosine, have been characterised in T. vaginalis. In the evaluation of the ADA profile in different T. vaginalis isolates treated with different iron sources or with limited iron availability, a decrease in activity and an increase in ADA gene expression after iron limitation by 2,2-bipyridyl and ferrozine chelators were observed. This supported the hypothesis that iron can modulate the activity of the enzymes involved in purinergic signalling. Under bovine serum limitation conditions, no significant differences were observed. The results obtained in this study allow for the assessment of important aspects of ADA and contribute to a better understanding of the purinergic system in T. vaginalis and the role of iron in establishing infection and parasite survival.

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Cultivation

Cited by 11 publications

References 35 publications

Analysis of the NTPDase and ecto-5'-nucleotidase profiles in serum-limited Trichomonas vaginalis

Analysis of the NTPDase and ecto-5'-nucleotidase profiles in serum-limited Trichomonas vaginalis

Pharmaceutical activity of a synthetic heterocyclic (C<sub>15</sub>H<sub>12</sub>N<sub>5</sub>OCl) compound on <i>Entamoeba histolytica</i> and <i>Giardia lamblia</i>

Modulatory effect of iron chelators on adenosine deaminase activity and gene expression in Trichomonas vaginalis

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