[ rapid method for extraction of water soluble beta[1,3] glucan from the cell wall of candida albicans]

In recent decades, beta-glucans have been used as important complementary and alternative medicines for numerous immunocompromised individuals and those with advanced cancer. The most active form of beta-glucans is beta[1,3]D-glucan and its most common source is cell wall of Candida albicans. Recently it has been introduced as a nano particle design to be used as a carrier for drug delivery. The current study researches a rapid method for the extraction of beta[1,3]D-glucans. The present study was conducted at Tarbiat Modares Medical University in 2012. Candida solubilized beta-glucans were obtained by oxidation of the cell wall with sodium hypochlorite and sodium hydroxide. The particle part could be solubilized by treatment with dimethylsulfoxide [DMSO] and zymolyase digestion to extract beta[1,3]Dglucan. The soluble fractions were lyophilized. We performed the Callose test to verify the presence of beta[1,3]D-glucans. Solubilized fractions were dissolved in D2O and 1HNMR spectra were measured. The soluble beta[1,3]D-glucan fraction which was derived from 1 g of dried Candida albicans germ tube weighed 190 mg. beta[1,3]D-glucan was verified by the Callose test and [1]H-NMR test compared with Curdlan [standard]. [1]H-NMR spectra verified the existence of beta[1,3]D-glucan in the final product. In the present study, extraction of beta[1,3]D-glucan by oxidation of the cell wall using sodium hypochlorite yielded more pure beta[1,3]D-glucans in comparison with other extraction methods. Thus it might represent a rapid method of extraction.

[Antifungal effect of zinc oxide nano_particlees of the standard strains of candida albicansbiofim growth on catheters]

Candida albicans is an important human pathogen that can causes systemic infections, predominantly among populations with weak immune systems. Candida spp. produces biofilms on synthetic materials such as catheter, which facilitates adhesion of the organisms to devices. Since the frequent replacement of these devices is difficult and expensive to achieve solutions for rapid identification and control of biofilm infections is essential. The purpose of this study is, the use of zinc oxide nanoparticles in the manufacture catheter and evaluation of its inhibitory effect in preventing of the biofilm formation. In this experimental study, the catheters in making poly vinyl chloride, Znonano particles with aconcentration 2.027 microg/ml was used. Biofilms were formed over a series of time intervals [2, 4, 6, 8, 24 and 48 h]. At each time interval, biofilm formation was measured with the XTT and biomass assay and concurrently assessed by light microscopy. Results showed that catheters containing zinc oxide increases with time in cell density [absorbance increase] were lower than those without catheters. Similar results were also measured in dry weight. This study demonstrated antifungal activity of ZnO. ther for biofilm formation on medical equipment such as catheters, implants, dentures, artificial heart valves, etc can be inhibited by ZnO nanoparticles

[Evaluation of antifungal activity of essential oils of Thymus vulgaris, Petroselinum Crispum, Cuminum cyminum and Bunium persicum on candida albicans in comparison with Fluconazole]

Nowadays notable increase in acquired resistance of Candida species to antifungal drugs and necessity of using agents with antifungal properties is unavoidable. In some plants due to presence of components such as polyphenols have antimicrobial properties. In this study antifungal effects of essential oils of Thymus vulgaris, Petroselinum crispum, Cuminum cyminum and Bunium persicum on standard strain of Candida albicans were evaluated. 25 grams leafs of the Thymus vulgaris and Petroselinum crispum and seeds of the Cuminum cyminum and Bunium persicum were dried and grinded after that the essential oils of each mentioned plant were prepared by Clevenger system. Serial dilutions of essential oils were made in 96 well microtiter plates. Minimum Inhibitory Concentration [MIC] and Inhibitory zone diameter were assessed by Microdilution broth and Disc diffusion agar techniques, MIC50, MIC90 and MFC were also determined. Minimum inhibitory concentration 90 [MIC90] essential oils of Thymus vulgaris, Petroselinum crispum, Cuminum cyminum and Bunium persicum were respectively 25, 72, 412, 130 micro g/ml and Minimal Fungicide Concentration [MFC] were 48, 146, 62, 280.Inhibitory zone diameters were 28, 20, 12, 15 millimetres. In this evaluation essential oils of Thymus vulgaris, Petroselinum crispum, Cuminum cyminum and Bunium persicum showed suitable antifungal effects against growth of standard strain of Candida albicans. Thus these herbal essences after supplementary studies possibly can be suitable substitute for chemical medicine on Candida infections especially on mucocutaneous Candidiasis

effects of candida albicans cell wall protein fraction on dendritic cell maturation

Candida albicans is a member of the normal human microflora. C. albicans cell wall is composed of several protein and carbohydrate components which have been shown to play a crucial role in C. albicans interaction with the host immune system. Major components of C. albican cell wall are carbohydrates such as mannans, beta glucans and chitins, and proteins that partially modulate the host immune responses. Dendritic cells [DC], as the most important antigen-presenting cells of the immune system, play a critical role in inducing immune responses against different pathogens. We investigated the effect of the cell wall protein fraction [CPF] of C. albicans on DC maturation. The CPF of C. albicans cells was extracted by a lysis buffer containing sodium dodecyl sulphate, 2-mercaptoethanol and phosphate-buffered saline. The extract was dialyzed and its protein pattern was evaluated by electrophoresis. Dendritic cells were purified from Balb/c mice spleens through a three-step method including mononuclear cell separation, as well as 2-h and overnight cultures. The purified CPF was added at different concentrations to DC. The purity and maturation status of DC were determined by flow cytometry using monoclonal antibodies against CD11c, MHC-II, CD40 and CD86. Treatment of DC with 10 micro g/ml of CPF increased the expression of maturation markers including MHC-II, CD86 and CD40 on DC compared to the control group. In this study we used C. albicans CPF with the molecular weight of 40-45 kDa for pulsing and maturation of dendritic cells. Since according to our results CPF significantly increased the expression of maturation markers on DC, we suggest that CPF may act as an efficient immunomodulator, or may be used as a potential adjuvant to boost the host immune system against infections

Effect of cell wall, cytoplasmic fraction and killed-Candida albicans on nitric oxide production by peritoneal macrophages from BALB/c mice

The fractions of Candida albicans have been used as an immunomodulator. The present work assessed the effect of different fractions of C. albicans on nitric oxide [NO] production by mice peritoneal macrophages. Cell wall and cytoplasmic fractions of C. albicans ATCC 10321 strain were extracted. Mice peritoneal macrophages were purified and cultured. Different concentrations of both fractions and also killed C. albicans cells were used for macrophages stimulation and evaluation of NO production. NO amount was detected in culture supernatants of macrophages by Griess reagent. Also, MTT assay was performed to assess the viability of macrophages. The results elucidated that suppressive effect of cell wall proteins on NO release was significant at the dose of 100 micro g/ml [P=0.01], while cytoplasmic fraction increased NO amount at the dose of 1 micro g/ml compared to the control group [P=0.003]. Augmentation of NO production was statistically significant at 200 killed C. albicans per well [P=0.006]. According to our findings, cytoplasmic fractions and killed C. albicans have a positive effect on NO production by peritoneal macrophages, while cell wall fractions did not. Therefore, it is proposed that C. albicans fractions can be studied more as inflammation modulators

[Cloning and sequencing of granular antigen 7 [GRA7] in TOPO vector and transformation in TOP10]

Toxoplasmosis, caused by an intracellular protozoan parasite, and the Toxoplasma gondii, is widespread throughout the world. In recent years, significant progress has been made in the identification of vaccine candidates which could induce a protective response. GRA7, an excretory 29 kDa Toxoplasma gondii a dense granular antigen released by infected host cells. In tachyzoite-infected cells, p29 accumulates within the parasitophorous vacuole and co-localizes with its delimiting membrane. In the present work, first genomic DNA of Toxoplasma gondii was extracted and used for amplifying of GRA7 gene as a template. Then PCR product was extracted from agarose gel and cloned into TOPO vector. The plasmid containing GRA7 gene was extracted from the transformed bacteria [TOP10 strain] and sequenced. Sequence analysis of GRA7 gene cloned into TOPO vector showed only one base difference when composed with the gene bank sequence for RH strain was only one base. The results indicated that this clone is suitable for subcloning in Prokaryotic and Eukaryotic plasmid

Seroepidemiological Survey of Hydatid cyst by ELISA in Kordestan Province

This study was conducted to evaluate the frequency of human hydatidosis in Kurdestan province by ELISA technique. In this study the sera of 1979 individuals were collected from different area [cities and villages] of Kordestan province. The serum dilution of 1/400 was selected for ELISA test. The results indicated that 1.12% of the individuals from Kordestan province showed positive sera. The results also showed that in Kordestan 0.9% and 1.42% of the people who live in the cities and villages had positive sera respectively. In this study 1.65% of female and 0.45% of male were positive. From the obtained result we found maximum number of infected people were in the range of 30-40 years [1.59%]. According to the results obtained from this study the highest percent of infection was found in the city of Ivandarre, the reason for this difference [1.69%] is due to the fact that most of the people who are involved in animal husbendary in the province live in this city