Do antidiabetic drugs prevent the transformation of Acanthamoeba trophozoite into cyst form?

This study examines the effects of three different drugs with metformin, acarbose and pioglitazone active ingredients used for antidiabetic purposes on Acanthamoeba cysts and trophozoites. Cultures of A. castellanii trophozoites and cysts were prepared to test the anti-amoebic activity of metformin, acarbose and pioglitazone. Cultures were then prepared for A. castellanii cyst and trophozoite forms and parasites were exposed to different concentrations (0.750 mg/mL, 0.375 mg/mL, 0.186 mg/mL and 0.093 mg/mL) of metformin, acarbose and pioglitazone. As a result of the study, the reproductive potential suppressive effects and conversion from trophozoite form to cyst form of all three substances on A. castellanii trophozoites and cysts were determined. Parasites were counted at 12, 24 and 48 hours in the cell counter after staining with trypan blue. In comparison of the effects of metformin, acarbose and pioglitazone used in the study on A. castellanii trophozoites and cysts, it was observed that all three substances were statistically effective against cysts and trophozoites at a concentration of 0.750 mg/mL. Furthermore, it was determined that all concentrations of the three active substances included in the study significantly decreased the rate of cyst formation even at the end of the 7th day. In this context, it was determined that all three substances have amebicidal effects, and they significantly inhibit the transformation of A. castellanii trophozoites to cyst form. It is thought that these active substances, which are currently used as anti-diabetic, can be used in combination with other drugs in A. castellanii infections based on our study findings.

Biogenic Nanocopper: Eco-Friendly Synthesis, Characterization, and Their Anti-Trichomonas vaginalis, Anticancer, and Antimicrobial Effects.

Biogenic nanocopper (BNC) agents exhibit strong anticancer, antimicrobial, and antiparasitic effects. Their fewer side effects to normal cells cause them to be preferred to treat various diseases. Metal nanoparticles, particularly copper nanoparticles, are attracting more significant interest as therapeutic agents with the improvement of green synthesis methods. Studies to reduce the side effects of copper nanoparticles to exhibit strong pharmacological properties are progressing intensively. Here, BNCs with reduced side effects were synthesized using L-ascorbic acid as the reducing agent and various concentrations of copper (II) chloride. BNCs exhibited significant pharmacological activity on cancer, bacteria, and Trichomonas vaginalis cells. The newly synthesized BNCs were characterized by scanning electron microscopy, UV-Vis spectrophotometry, Fourier transform infrared spectroscope, and Differential/Thermal Gravimetric Analysis. The pharmacological activity of BNCs was evaluated by obtaining their inhibitor concentration and minimum inhibitory concentrations against some cancer, bacteria, and T. vaginalis cells. Newly synthesized BNCs have various shapes such as cubic, spherical, or rod and particle size distribution between 70 and 100 nm. According to experiment results, the newly synthesized BNCs were a significantly antiproliferative, antibacterial, and anti-T. vaginalis effect on cells compared to the control drugs. These findings confirm newly synthesized BNCs and their in vitro pharmacological potential. Further research should be targeted on the preclinical study of absorption, distribution, metabolism, excretion/toxicity (ADME/Tox) and in vivo effects on cancer and microbial pathogens.

Combination of infra-red light with nanogold targeting macrophages in the treatment of Leishmania major infected BALB/C mice.

PURPOSE: In the treatment of cutaneous leishmaniasis (CL), developing drug resistance, existing toxic effects of drugs and failure respond to treatment cause the need to try different treatment methods. We investigated the effect of gold-conjugated macrophage-specific antibody on amastigotes under infra-red light for the treatment of CL. METHODS: Female BALB/c (4-8 weeks old, 20 ± 5 g weight) mice were used in the study. The L. major strain was inoculated on the soles of mice in amastigote form and subpassed. Nanogold (Au), Au + macrophage-specific antibody (MSA) modification and near infra-red (NIR) (5 seconds) were applied to mice groups that developed cutaneous leishmaniasis on their soles. On the 5th and 10th days of the treatment, the lesions were examined clinically and pathologically. RESULTS: When the erythema values were examined, the highest decrease was calculated in the Au + MSA + NIR group in the measurements made on the 10th day (p < 0.014). The best improvement in 10th day measurements is in the NIR and Au + MSA + NIR groups when area values were examined (p = 0.011, p = 0.001). There was a statistically significant difference between the groups in terms of parasite load (PL) (p < 0.005) in pathological evaluation. According to PL grouping, the best result is NIR (p = 0.002). When both main titles (clinical and pathological) are examined, the Au + MSA + NIR group is thought to have an optimal therapeutical feature. CONCLUSIONS: Au + MSA + NIR combination could be a new treatment approach for CL treatment.

The amoebicidal activity of three substances derived from benzothiazole on Acanthamoeba castellanii cysts and trophozoites and its cytotoxic potentials.

Acanthamoeba species are free-living amoebae isolated from many ecological areas such as swimming pools, dams, lakes, soil, and air filters. These amoebae are usually causing granulomatous amebic encephalitis and amebic keratitis in immunosuppressive individuals. In this study, the reproductive potential and morphological changes determined of Acanthamoeba castellanii trophozoite and cyst forms exposed to three different active substances derived from benzothiazole. Furthermore, the cytotoxic potential of these active substances determined by XTT analysis. In the study, axenic cultures prepared for Acanthamoeba castellanii cyst and trophozoite forms and parasite exposed to different concentrations of active substances. Cell counts of parasite cultures were performed at the 30 minutes, 1st, 6th, 12th, 24th, and 48th hour periods. As a result of the study, the reproductive potential suppressive effects of all three substances on Acanthamoeba castellanii trophozoites and cysts were determined. The most effective of these substances was 2-Amino-6(trifluoromethoxy)-benzothiazole. In the first three concentrations of this substance (0.1%, 0.05%, 0.025%), no determined trophozoite and cysts at the end of twenty four. Due to its strong ameobicidal effect, it is thought that 2-Amino-6(trifluoromethoxy)-benzothiazole may be a new therapeutic agent in diseases caused by acanthamoeba parasites by supporting this study with animal experiments.

A model Organism for Antihelminthic Activity: Caenorhabditis elegans and Nigella sativa

Purpose: In our study, antihelminthic activity of Nigella sativa was tested on a model organism Caenorhabditis elegans (C. elegans). Methods: N2 (wild type) C. elegans and Escherichia coli OP50 were purchased from the University of Minnesota, Ceanorhabditis Genetic Center for the study. C. elegans were grown in NGM (Nematode Growth Medium) solid culture medium. After synchronization, nematodes in adult form were exposed to Nigella sativa seed oil at concentrations of 1%, 0.1%, 0.01%, 0.001% and 0.0001%. The study continued for 18 days until all nematodes in the experimental and control groups died. Alive and dead nematodes were recorded every day. The nematodes in which pharengeal pumping stopped was recorded dead. Result: According to our findings, all nematodes died at the end of the fourth day at a concentration of 1%. In addition, no alive nematod was observed at the end of the fifth day at concentration of 0.1%; whereas at the end of the 13th day all nematodes died at concentration of 0.01%. When the data were analyzed statistically, the difference between at concentrations of 0.1%, 0.01%, 0.001% and the control group was significant (p<0.05). Conclusion: The fact that helminths are usually macroscopic in size and most of them cannot be produced in the laboratory by culturing is a major obstacle for scientific studies. Our study has shown that C. elegans can be a good model in studies of antihelmintic activity and that Nigella sativa has an anthelmintic effect.

A new therapeutic approach for the treatment of cutaneous leishmaniasis: Photothermal application of macrophage-specific antibody binding graphene oxide nanoparticles.

BACKGROUND & OBJECTIVES: Cutaneous leishmaniasis (CL) is a zoonotic and anthropogenic protozoal disease. We aimedto develop a new therapeutic approach for the treatment of CL. METHODS: BALB/c mice have infected L. major amastigotes from their footpads. Twenty-one days later after injection, the animals were divided into three control and three experimental groups. The intralesional injection of graphene oxide and photothermal application (GO+PA) were applied to the first experimental group (Group 1); graphene oxide modified with a macrophage-specific antibody and photothermal application (MSA+GO+PA) were applied to the second experimental group (Group 2), and the photothermal application (PA) was applied to the third experimental group (Group 3). Miltefosine was administered orally to the first control group (Group 4); the second control group that is not treated was assigned as the positive control (Group 5) and the third control group was assigned as the negative control (Group 6). Lesions were examined (erythema and edema) after the 5th day and 10th of the treatment, clinically. On the 10th day of the treatment, the level of TNF-α, IL-1, IL-6, and IFN-ɤ were detected histopathologically and immunohistochemically. RESULTS: In the 5th day of the treatment it was observed that 50% of the animals were completely treated with Group 2, and in the 10th day, the ration raised to 75%. INTERPRETATION & CONCLUSION: We showed a novel application to treat CL by using MSA modified GO and PA within 10 days. According to our study outcomes, this application could be a new treatment approach for CL cure.

Amoebicidal activity of benzothiazole on Acanthamoeba castellanii cysts and trophozoites and its cytotoxic potentials.

Acanthamoeba spp. are free-living amoebae isolated from many ecological areas such as swimming pools, dams and lakes, and soil. Granulomatous amebic encephalitis and amebic keratitis, caused by Acanthamoeba spp., usually occurs in chronically ill, debilitated individuals, in immunosuppressed patients and treatment is quite difficult. This study aimed to determine the effect of benzothiazole on trophozoite and cyst forms of Acanthamoeba castellanii (A.castellanii). Axenic cultures of A. castellanii trophozoites and cysts were prepared to test the amoebicidal activity of benzothiazole. The concentrations of benzothiazole in 24-well plates were prepared as 0.08%, 0.04%, 0.02%, 0.01%, 0.005%, and A. castellanii cysts and trophozoites were added to these cultures. Parasites were counted at 0, ½, 1, 3, 6, 12, 24 and 48 h in the cell counter after staining with trypan blue. Cytotoxicity of benzothiazole on the WI-38 human fibroblast cell line was also tested. Between 0.08% and 0.01% concentrations of benzothiazole showed a strong amoebicidal activity at 24 and 48 h. A significant decrease in 0.005% concentration in the number of live trophozoites and cysts was detected between 6 and 48 h. As a result of the cytotoxicity studies, benzothiazole did not show any cytotoxic effect on the WI-38 human fibroblast cell line even at 1% concentration. Benzothiazole could be concluded as a new therapeutic agent against Acanthamoeba. On the other hand, in vivo studies are needed to confirm the efficacy of the biological effect.

The physiological and lifespan alterations in Caenorhabditis elegans exposed to different dosages of melatonin.

Aging is a process that begins at birth and ends with death. This process is accompanied by environmental effects, which cause structural and functional changes in cells and tissues. With regards to healthy aging, melatonin significantly extends lifespan. This study aims to show the anti-aging effects of melatonin on lifespan by a model organism called Caenorhabditis elegans. The nematode strain N2 (wild-type) was acquired, and E. coli OP50 was used in the study. Worms were grouped into a control group (n=100), and six experimental groups (group 1, 2, 3, 4, 5 and group 6) (n=100 in each of them). Interventions were made by exposing Caenorhabditis elegans to various dosages of melatonin and follow up was made for 21 days. The survey of Caenorhabditis elegans, which depends on time and dosage as the main outcome measures, was examined microscopically. Different dosages of melatonin affected the lifespan and morphology of Caenorhabditis elegans. Melatonin might be used in the prevention of aging.

Comparison of Reproduction Densities in Different Liquid Media of Trypanosoma cruzi and Cryopreservation.

OBJECTIVE: This study aims to determine the optimum liquid medium for the reproduction of Trypanosoma cruzi strains and provide cryopreservation. METHODS: The reproduction density of T. cruzi strain was evaluated in the following five different commercial liquid culture media: RPMI 1640, Medium 199 (M199), Schneider's Insect Medium (SIM), Nutrient Broth (NB), Brain Heart Infusion Broth (BHIB). Cultures were monitored on every other day for a period of 24 days. Cryopreservation of T. cruzi was also performed and viability was tested after six months. RESULTS: Epimastigotes of T. cruzi were not found to be produced in NB and BHIB media. Significant difference was not observed among the reproduction potential of RPMI-1640, M199, and SIM after evaluating the data for the first 10 days. Between days 12 and 24, RPMI-1640 was found to be the best reproduction medium. From the 18th day onwards, parasites transformed amastigotes. On the 24th day, the highest level of amastigote amount was observed, and reproduction was determined to have stopped. As a result of cryopreservation, it was determined that the survival of T.cruzi continued after six months.. CONCLUSION: Thus, the selection of RPMI-1640 medium, followed by M199 and SIM media would be appropriate when studying T. cruzi epimastigotes. Studies using epimastigotes should be planned for up to 18 days and for those using amastigotes, it would be appropriate to plan the studies after the 18th day. Moreover, T. cruzi can be cryopreserved with 15% DMSO and stored for a long time in liquid nitrogen.

The Effects of Erzincan Grape (Vitis vinifera spp., Cimin) and Benzothiazol on a Caenorhabditis elegans Organism Model.

BACKGROUND: Grapes and their products are known to have been used for the treatment of diseases throughout history. OBJECTIVE: It was aimed to investigate the effects of Erzincan Cimin grapes on an organism model of Caenorhabditis elegans N2 wild type and C. elegans BS913 strains with gonad cancer. MATERIALS AND METHODS: The effects of methanol extracts of the skin and seeds of Erzincan Cimin grapes were examined separately on C. elegans N2 wild type and an effect was determined on lifespan. By applying GS-MS analysis, a potential agent substance was determined in the skin and seed methanol extracts. This substance was purchased and the effects of this substance were investigated on lifespan and fertility in C. elegans BS913 strains with gonad cancer. In addition, the effects on young subjects exposed to this agent substance in L1 form were investigated. RESULTS: Grape seed and skin methanol extract was observed to prolong the lifespan most at a dose of 10 mg/100 mL. Lifespan was determined to be at a maximum in a gonad cancer organism model with benzothiazol at a dose of 50 ppm. At the same dose, positive effects were determined on the fertility of strains with cancer. When the effects of benzothiazol were examined on young L1 forms, an evident retardation of growth was determined at doses of 10, 50, and 100 ppm. CONCLUSION: Owing to anti-carcinogenic effects of benzothiazol and benzothiazol-derived substances, they can be considered as agent substances in academic studies related to cancer. SUMMARY: The effects of methanol extracts of the skin and seeds of Erzincan Cimin grapes were examined on C. elegans N2 wild type and an effect was determined on lifespanThrough GS-MS analysis, benzothiazol was determined in the skin methanol extractsBenzothiazol was purchased and the effects of this substance were investigated on lifespan and fertility in C. elegans BS913 strains with gonad cancerThe effects on young subjects exposed to benzothiazol in L1 formGrape seed, skin methanol extract, and benzothiazol was observed to prolong the lifespanPositive effects were determined on the fertility of gonad cancer strains. Abbreviations used: GC-MS: gas chromatography and mass spectrometry; C. elegans: Caenorhabditis elegans; NGM: Nematode growth medium; E. coli: Escherichia coli; FUDR: Fluorodeoxyuridine; LDL: Low-density lipoprotein.