[Evaluation of anti tumor activity of candida albicans cell wall fraction on human fibrosarcoma cell in vitro]

Matrix metalloproteinases [MMPs] plays a role in the several physiologic and pathologic events. There is some evidence indicating the involvement of MMPs in tumor invasion and inflammatory diseases. In this study we evaluated the effect of cell wall Candida albicans fraction on MMP production by the fibrosarcoma cell line using an in vitro cytotoxicity assay, sodium dodecyl sulfate-polyacrylamide, and gelatin zymography. It was an experimental study. Fraction of cell wall Candida albicans was extracted by lysis buffer and] 0, 10, 50, 100 [micro g/ml from fraction was added to human fibrosarcoma cell line in 96 well microplate and inhibitory effect on tumor cell invasion was investigated by MTT and zymography test. Our results showed that fraction of cell wall Candida albicans was found to exhibit a inhibitory effect on tumor cell line in MTT test because the viability of tumor cells was decreased versus the control groups. Also the production of metalloproteinases enzyme in presence of fraction was diminished significantly [P<0.05]. Since inhibition of MMP activity has been employed in modality therapy in diseases such as cancer, this fraction could be promised in the preparation of anti-MMP therapeutic derivatives

[Shigella dysentery stxA mutant [R170L-A231D-G234E] gene design and optimization of recombinant protein expression and purification]

Background and aims: Shigella dysentery is one of the most important human pathogenic intestinal bacteria. Entrance of the shigella toxin into the epithelial cells inhibits protein synthesis leading to cell death. In spite of great investigations on vaccine production against S. dysentery, studying to achieve significant stxA recombinant protein still remains important. The objective of this study was to determine the appropriate mutation loci and designing stxA subunit synthetic gene, its further expression and optimization and ultimately assaying purification method for further immunization studies

Methods: Three mutant stxA gene including [R170L-A231D-G234E] were designed and the synthetic gene in pET28a plasmid was obtained and confirmed by PCR. Thereafter the plasmid was transformed into the host cell E.coli BL21 DE3 after which gene expression was optimized and protein purity assay was then performed

Results: Preliminary studies led to mutant stop gene design, after which it was confirmed by synthetic plasmid and PCR. Expression and optimization were then performed which resulted in large amount of protein inclusion bodies. Purification of inclusion bodies and protein which resulted solubilization was done with a combinatorial method

Conclusion: With regard to the mechanism of shiga toxin effect and favorable mutation design with new arrangement, less toxicity of expressing protein is predicted than previous other mutants, posing a better vaccine candidate

Evaluation of Bax encoding plasmid for increasing efficacy of DNA vaccine plasmid encoding gB of Herpes Simplex virus type 1

Evaluation of Bax encoding plasmid for increasing efficacy of DNA vaccine plasmid encoding gB of Herpes Simplex Virus type 1. Materials and Methods: We compared three different dosages of Bax encoding plasmid [pcbax] including 10, 25 and 50 ?g of plasmid DNA. They were co-injected ineradermally with glycoprotein B [gB] of herpes simplex virus [HSV]-1 encoded plasmid [pcgB] in C57BL/6 mice to elicit immune responses to protect against lethal HSV-1 challenge. Immune responses to the antigen were assessed by lymphocyte proliferative responses and cytokine [INF-gamma and IL-4] release assays. The study demonstrates that the mice immunized with 25 micro g pcbax together with pcgB have more efficient protection than the mice immunized with 10 and 50 micro g of pcbax and pcgB. Analysing of cell-mediated responses show that the mice immunized with 25micro g pcbax and pcgB induce stronger lymphocyte proliferative responses and higher levels of INF-gamma and IL-4 compared to the mice are received 10 and 50 micro g of pcbax and pcgB. The data show that co-immunization with 25 micro g of pcbax and pcgB increase immune responses compared to 10 and 50 micro g of pcbax and pcgB. This can be considered a promising approach for development an efficient DNA vaccine against HSV-1 or other pathogens

immunomedulatory effect of Ganoderma Lucidum polysachharide extract on BALB/c peritoneal macrophage function

Ganoderma Lucidum has been regarded as a natural immunomodulator. The exact carbohydrate epitope responsible for the immunomodulatory activity and its receptor have not been identified, but it seems likely that it is the receptor CR3 [complement receptor 3] which can bind to beta-glucan polysachharide. Because glucose-6-phosphate dehydrogenase [G6PD] activity has a critical role in the regulation of macrophage functions such as nitric oxide [NO] production, we assessed the immunomodulatory effect of GL-PS in BALB/c peritoneal macrophages. For this purpose, BALB/c mice peritoneal macrophages were isolated and treated with various concentrations of GL-PS [0.001, 0.01, 0.1, 1, 10, 100 microg/ml]. After 24 hours, the viability of treated macrophages was measured by MTT assay at 540 nm and the effective dose was determined to be 0.1 microg/ml. Then, macrophages were sonicated and special activity of G6PD was measured in the cell extracts by measuring the alterations in NADPH absorption at 339nm and protein concentration by Bradford method. Also, NO production was determined by use of Griess-reagent after 18 hours. Results of this study showed that 0.1 microg/ ml of GL-PS had the maximal effect on cell viability [stimulation Index] in comparison to other doses [0.05

Evaluation of the susceptibility of dermatophytes to garlic extract

In this study, we evaluated the activity of garlic extract against Dermatophytes which is planned to be used instead of chemical durgs. Garlic has been widely used as medicine since ancient times for varieties of illnesses, including abdominal pain, parasitic infection, insect and snakebites, hemorrhoid and rheumatism. In the last decades, garlic has been reported to display antibiotic and antifungi activities. Garlic was obtained from Hamadan, Iran. Using the Mantis method, dry garlic bulbs were peeled and homogenized with two parts of distilled water in a blender and liquid garlic extract was obtained. Then the homogenized garlic extract was run through Amicon DIAF10 ultra-filtration system, using XM and PM membranes. The ultra-filtrated fractions were collected as Residue [R] 300, 100, 50, 30, 10 and filtrate [F] 10. The fractions were evaluated by SDS-PAGE, using 14 percent Acrylamide gel. Serial dilutions of fraction from 1/2 up to 1/32 were tested against each Dermatophyte in Sabourauds Dextrose agar and Minimum Inhibition Concentration [MIC] was obtained. The Dermatophytes tested included: Trichophyton mentagrophytes var. mentagrophytes, Triochophyton Mentagrophytes var. interdigitale, Tricophyton rubrum, Tricophyton tonsurans, Microsporum canis, Microsporum gypseum and Epidermophyton floccosum. An ointment was prepared with Fraction F10 as active ingrediant and was used for treatment of dermatophitosis of ginea pigs. The result showed that F10 inhibited growth of Microsporum canis, Epidermophyton floccosum, Tricophyton rubrum, Tricophyton tonsurans. MIC 1/4 was active against Microsporum gypseum and 1/2 against Triochophyton Mentagrophytes var, interdigitale. Trichophyton mentagrophytes var. Mentagrophytes was resistant to all dilutions. lso the ointment used for treatment of dermatophitosis of guinea pig showed a statistically significant inverse relation between the severity and diameter of lesions and the duration of treatment [p<0.01]. This research showed that F10 fraction, which contains nonprotein components, is the most effective treatment for dermatophitosis