Background:Systemic candidiasis is a major public health concern. In particular, in immunocompromised people, such as patients with neutropenia, patients with Acquired Immune Deficiency Syndrome (AIDS) and cancer who are undergoing antiballistic chemotherapy or bone marrow transplants, and people with diabetes. Since the clinical signs and symptoms are nonspecific, early diagnosis is often difficult. The 65-kDa mannoprotein (MP65) gene of Candida albicans is appropriate for detection and identification of systemic candidiasis. This gene encodes a putative b-glucanase mannoprotein of 65 kDa, which plays a major role in the host-fungus relationship, morphogenesis and pathogenicity.Objectives:The current study aimed to identify different species of Candida (C. albicans, C. glabrata and C. parapsilosis) using the Polymerase Chain Reaction (PCR) technique and also to evaluate C. albicans MP65 gene expression in BALB/C mice.Materials and Methods:All yeast isolates were identified on cornmeal agar supplemented with tween-80, germ tube formation in serum, and assimilation of carbon sources in the API 20 C AUX yeast identification system. Polymerase Chain Reaction was performed on all samples using species-specific primers for the MP65 65 kDa gene. After RNA extraction, cDNA synthesis was performed by the Maxime RT Pre Mix kit. Candida albicans MP65 gene expression was evaluated by quantitative Real-Time (q Real-Time) and Real-Time (RT) PCR techniques. The 2-ΔΔCT method was used to analyze relative changes in gene expression of MP65. For statistical analysis, nonparametric Wilcoxon test was applied using the SPSS version 16 software.Results:Using biochemical methods, one hundred, six and one isolates of clinical samples were determined as C. albicans, C. glabrata and C. parapsilosis, respectively. Species-specific primers for PCR experiments were applied to clinical specimens, and in all cases a single expected band for C. albicans, C. glabrata and C. parapsilosis was obtained (475, 361 and 124 base pairs, respectively). All species isolated by culture methods (100% positivity) were evaluated with PCR using species-specific primers to identify Candida species. Relative expression of Mp65 genes increased significantly after C. albicans injection into the mice (P < 0.05).Conclusions:The results of the current study showed that the PCR method is reproducible for rapid identification of Candida species with specific primers. Mp65 gene expression of C. albicans after injection into the mice was 2.3 folds higher than before injection, with this difference being significant. These results indicated that increase of Mp65 gene expression might be an early stage of infection; however definitive conclusions require further studies.
HSV-1 is associated with oral lesions. Recently, anti-herpetic activity of different plant species has been investigated. In this study, the effects of Artemisia aucheri aqueous extract on the HSV-1 virus-infected Vero cells were assessed. The highest cell viability occurred in plant aqueous extracts was with a concentration of 75 μg/mL, 1–2 h before viral infection. The IC50 of the aqueous extract of 24.7 μg/ml was calculated. Most percentage of infected cell inhibition (89.6%) was with the chloroform fraction in concentration of 75 μg/ml, and the least percentage of infected cell inhibition (21.7%) was in concentration of 12.5 μg/ml with the ethyl acetate fraction in comparison with untreated control. Moreover, Q-PCR results revealed that the expression of genes UL46 and US6 were significantly reduced in the presence of different treatments utilized in the experiment. In conclusion, the present study proposes that aqueous extracts of medicinal plant Artemisia aucheri have anti-viral property and may be considered as a remedy for HSV-1 treatment.
Background;: Leishmaniasis is one of the major diseases caused by the intracellular parasite of Leishmania. It has become one of the most dangerous health problems today. Our aim of the present study is to compare the effects of Sambucus ebulus leaf and fruit extracts on Leishmania major in vitro. Methods: In this study, we used MTT, promastigote and amastigote assay to evaluate the effect of different concentrations of the extract on parasite and we compared their effects. The flow cytometry technique was also used to detect the apoptotic effect of the extracts on promastigotes. Results: According to MTT experiment IC50 concentration of leaf and fruit extracts on parasite was 157 μg/ml and 265 μg/ml, respectively. After analysis by flow cytometry, leaf and fruit extracts also showed apoptosis effect. Leaf and fruit extract caused 40.2 and 2.67 percent apoptosis. Conclusion: Based on the above assessment, we determined that the S. ebulus leaf extract has a more toxic effect on promastigotes and amstigotes than its fruit extract and maybe in the future that be used as a drug candidate.
Background: Nowadays, the use of herbal medicines in the prevention and treatment of diseases has been increased in the world, especially in Iran. Developing chemical drugs resistance in some Candida species and their side effects, conducting research to find new resources, especially medicinal plants, are of prime importance.
Aim: Acanthamoeba cause dangerous diseases in humans such as encephalitis and keratitis as an opportunistic pathogen. Due to the antioxidant, antimicrobial, antifungal, anti-Acanthamoeba and anti-leishmania activities of Artemisia, the aim of this study is investigate the effect of aqueous extract of Artemisia aucheri seed on Acanthamoeba trophozoites and cysts in vitro. Materials and Methods: Acanthamoeba trophozoites and cysts were propagated in appropriate culture medium. Aqueous extract of Artemisia aucheri were prepared at concentrations of 2000, 1000, 500, 250, 125 and 62.5 μg/ml and were added to both protozoa forms (trophozoites and cysts). Then, three techniques including trypan blue, MTT and flowcytometry were used to investigate the effect of this extract on Acanthamoeba trophozoites and cysts. Results: It was found that increasing the time and concentration of aqueous extract of Artemisia aucheri seed significantly reduced the number of live Acanthamoeba trophozoites and cysts (P ≤0.05). At the concentration of 2000 µg/ml the number of live trophozoites was 0% and at the concentration of 62.5 µg/ml the number of live trophozoites was 57.7%. Conclusion: The results of this study showed that the aqueous extract of Artemisia aucheri has anti-acanthamoeba activity and seems to have beneficial pharmacological effects on some diseases and complications caused by Acanthamoeba. Further research is needed to determine this issue.
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