Antiviral effect of Artemisia aucheri aqueous extract on UL46 and US6 genes of HSV-1.

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.

Evaluation of the Effects of Rumex obtusifolius Seed and Leaf Extracts Against Acanthamoeba: An in vitro Study.

BACKGROUND: Acanthamoebiasis treatment is a major and challenging problem due to the presence of resistant cyst form. Many herbal extracts and their derivatives have been used against trophozoites and cysts of Acanthamoeba, but no effective therapeutic agent has yet been discovered. Therefore, the present study aimed to evaluate the effect of Rumex obtusifolius (R. obtusifolius) extracts against a clinical strain of Acanthamoeba genotype T4 in vitro. METHODS: In this experimental study, after genotyping the clinical isolate, the hydroalcohlic extracts of R. obtusifolius seeds and leaves were prepared. Different concentrations (1.25, 2.5, 5 and 10 mg/ml) of extracts were tested in triplicate (24, 48 and 72h) on trophozoites and cysts of Acanthamoeba. The mortality of the parasite was assessed by trypan blue vital staining and flow cytometry analysis. RESULTS: Results showed that the extract of R. obtusifolius leaves at the concentration of 10 mg/ml killed 100% of trophozoites and cysts after 72 h. However, the seed extract of R. obtusifolius had weak inhibitory effects on trophozoites and cysts of Acanthamoeba. In the presence of 10 mg/ml of hydroalcoholic seed extract of R.obtusifolius in culture medium after 72 h, 28.6% of trophozoites and 0% of cysts of Acanthamoeba were killed. After analysis by flow cytometry, seeds and leaves extract indicated apoptosis effect. Seed and leaf extracts caused 2.6% and 0.4% percent apoptosis. CONCLUSION: These extracts are not promising candidates for further medicine development on acanthamoebiasis. Nonetheless, further research is necessary to clarify the effects of effective fractions of seed and leaf extracts of R. obtusifolius and their mechanisms of action.

Comparison of the Effects of Sambucus ebulus Leaf and Fruit Extracts on Leishmania major In Vitro.

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 the 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.

Anti-Candida and antioxidant activities of hydroalcohlic extract of Rumex obtusifolius leaves.

Due to increasing the use of antifungal drugs, the development of resistance in some Candida species and the consumption of the side effects of chemical drugs, use of new resources, especially medicinal plants are very important. The aim of this study was to investigate of anti-Candida and antioxidant activities of hydroalcohlic extract from leaves of Rumex obtusifolius. The Rumex obtusifolius Leaves were extracted using Ethyl acetate; methanol and distilled water (6:3:1) by Sox helet system. The hydroalcoholic extraction of Rumex obtusifolius was evaluated for their antioxidant capacities using in vitro methods; including 1-diphenyl-2-picrylhydrazyl radical scavenging, ß-Carotene bleaching test and reducing power assay. Total free phenolics, total flavonoids content and as well as the antifungal activity were also examined. The components of extract were analyzed via GC-Mass instrument. The extract was screened against 40 isolated pathogenic Candida species such as C. albicans and C. glabrata through agar diffusion method. The hydroalcoholic extract can strongly scavenge DPPH radical and its antioxidant capacities which are high correlated with the total free Phenolics and total flavonoids. Also, the extract had high capability inhibition of linoleic acid oxidation and the reducing ability. This study revealed a higher antioxidant capacity in the leaves of Rumex obtusifolius compared with control groups. The minimum inhibitory concentration values within 24 and 48 hours were 200-250µg/µL for C. albicans and 250µg/µL for C. glabrata. The extract includes high amounts of phenolic compounds and antioxidant activity showing is significant. Also, the results confirmed that leaves extract had a potential in anti-Candida activity and suggesting that it could be utilized as a potential source of herbal medicine drugs and natural antioxidants to prevent diseases associated with free radical, anti-fungal disease and food preservation.

A Study on the Association between CCRΔ32 Mutation and HCV Infection in Iranian Patients.

BACKGROUND: Mutations in the coding region of the Chemokine Receptor 5 (CCR5) genes reduce or eliminate CCR5 expression in immune cells and progression of HCV infection. This study aimed to investigate the role of this mutation in HCV infection in Iranian patients in comparison with healthy individuals. METHODS: 100 HCV infected patients and 100 healthy individuals were randomly selected. The CCR5Δ32 genotypes were determined using specific primers and PCR method. RESULTS: The agarose gel electrophoresis showed a189-bp fragment from wild type for both alleles of CCR5 gene. The CCR5-Δ32 allele was not found in any HCV infected and healthy subjects. CONCLUSION: The mutation in CCR5 gene was not detected in any of the two groups; therefore, the role of CCR5 gene expression in immune cells and progression of HCV infection needs to be studied in larger samples in our country.

Identification of Candida Species Using MP65 Gene and Evaluation of the Candida albicans MP65 Gene Expression in BALB/C Mice.

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.