Background: Acanthamoeba keratitis develops by pathogenic Acanthamoeba such as A. palestinensis. Indeed this species is one of the known causative agents of amoebic keratitis in Iran. MANNOSE BINDING PROTEIN ((MBP)) is the main pathogenicity factors for developing this sight threatening disease. We aimed to characterize (MBP) gene in pathogenic Acanthamoeba isolates such as A. palestinensis.Methods: This experimental research was performed in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran during 2007-2008. A. palestinensis was grown on 2% non-nutrient agar overlaid with Escherichia coli. DNA extraction was performed using henol-chloroform method. PCR reaction and amplification were done using specific primer pairs of (MBP). The amplified fragment were purified and sequenced. Finally, the obtained fragment was deposited in the gene data bank.Results: A 900 bp PCR-product was recovered after PCR reaction. Sequence analysis of the purified PCR product revealed a gene with 943 nucleotides. Homology analysis of the obtained sequence showed 81% similarity with the available (MBP) gene in the gene data bank. The fragment was deposited in the gene data bank under accession number EU678895 Conclusion: (MBP) is known as the most important factor in Acanthamoeba pathogenesis cascade. Therefore, characterization of this gene can aid in developing better therapeutic agents and even immunization of high-risk people.

Background: Dermatophytosis is common cutaneous fungal disease with worldwide distribution. Interleukin8 (IL-8) realized from keratinocytes in the presence of dermatophytic antigens causes induction of acute responses in dermatophyte infection and subsequently production of acute phase PROTEINs occurs in hepatocytes. C-reactive PROTEIN (CRP) and MANNOSE-BINDING lectin (MBL) are acute phase PROTEINs. Since few researches in the case of acute phase PROTEINs in dermatophytic infections has been accomplished, this study has been designed for determining serum CRP and MBL levels in patients affected to dermatophytosis.Methods: This was a cross sectional study and samples were carried out on 96 healthy individuals and 105 patients affected to dermatophytosis with non probable and in access procedure. For isolation and identification of dermatophyte direct microscopic examination, culturing and complementary examinations were done and for determination of serum CRP and MBL levels in healthy individuals and in patients ELISA test were used. For investigation of relevance between variables, Chi-square, Fisher exact, Mann-Whitney and Roc curve analysis were used and p< 0.05 was considered as meaningful level.Results: The median serum CRP level in healthy individuals and in patients group was 3.31±3.32 mg/ml and 16.60±35.96 mg/ml (p<0.001) respectively and the median serum MBL level was 1.53±1.87 mg/ml and 1.97±2.03 mg/ml (p=0.039) respectively. CRP (p<0.001) and MBL (p=0.042) were determined meaningful parameters for dermatophytosis. MBL deficiency (MBL concentrations <1 mg/ml) was higher in control subjects (56.2%) than in patients (41.0%). Conclusion: Findings of this study indicate increased concentrations of CRP and MBL in patients affected to dermatophytosis and their role in this infection. Probably observation of high frequency of MBL deficiency in healthy individuals in compare with patients group indicates that it is not predisposing factor in affecting to dermatophytosis.

Purpose: A lectin-like PROTEIN from the mushroom Agaricus bisporus has been shown to slightly increase the proliferation of RAW 264. 7 cells. Following its identification as a MANNOSE-BINDING lectin, henceforth called A. bisporus MANNOSE-BINDING PROTEIN (Abmb), the PROTEIN is hypothesized to stimulate the innate immune cells response. The present work was aimed to substantiate that hypothesis. Furthermore, this study complements Abmb exploration as a potential agent for anti-breast cancer, which its treatment is hampered with compromised immunity of patient receiving chemotherapy. Method: Abmb’s effect on the phagocytic activity of the macrophage was measured with FACS. Nitric oxide (NO) production was checked using Griess test while expression of the cytokines in the RAW 264. 7 cells was analysed at gene and PROTEIN level using polymerase chain reaction (PCR) and FACS, respectively. Abmb’s effect on the expression of surface markers of the human immune cells in the peripheral blood mononuclear cells (PBMCs) was checked with specific antibodies for targeted cluster differentiation (CD) and analysed using FACS. Results: Abmb increased the phagocytic activity of the macrophage and NO production. Abmb increased the expression of cytokines i. e. tumour necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10. With the PBMCs, Abmb activated dendritic and natural killer (NK) cells, but not the B-or T-cells. Conclusion: Abmb increased the activity of the macrophage cells and activated the immune cells that are related to the innate immune system, particularly the cellular immunity.

Objective: CREB1 is an important downstream PROTEIN for many signaling pathways. By designing efficient siRNAs against CREB1, it may be possible to assess the role of molecules involved in signaling pathways in different cell types. In this research the efficiency of CREB1 knockdown by two different siRNAs in K562 cells has been studied. Materials and Methods: siRNAs have been designed according to the criteria suggested by Reynolds et al. K562 cells were transfected by siRNA using Lipofectamine 2000. The efficiency of CREB knockdown has been assessed by quantitative relative Real-time PCR. Results: Our results have shown that only one of the siRNAs has a high level of inhibitory effect on CREB1 gene expression. The expression of CREB1 by this siRNA was knocked-down by 87% in K562 cells. Conclusion: In this research, although two siRNAs were designed according to the Reynolds et al. criteria, only one showed an inhibitory effect. Reasons other than the aforementioned criteria may be involved in effectiveness of siRNAs.

Introduction & Objective: Acanthamoeba is free-living amoeba that is found in soil, water, air as well as in human pharynx. Acanthamoeba is causative agent of granulomatose amoebic encephalitis (GAE) in immunosuppressed and AIDS individuals and amoebic keratitis in people who use the lens. Pathogenic species of Acanthamoeba have PROTEIN receptors named MANNOSE BINDING PROTEIN ((MBP)). Acanthamoeba via (MBP) adhere to the glycoPROTEINs included MANNOSE. Acanthamoeba adhesion to the target cells induces a protease secretion is called MANNOSE inducing PROTEIN-133 (MIP-133). Exogense MANNOSE can inhibit the adherence of Acanthamoeba; also, it can increase the cytopathatic effect (CPE) through increase the secretion of MIP-133. In the present work, the effect of monosaccharides on the virulance of Acanthamoeba isolated from patient with amoebic keratitis, in HeLa cell culture was investigated.Materials & Methods: The isolates were cultured in HeLa cell culture, then 100, 50, 10, 1 and 0.1 mM of galactose, glucose and MANNOSE were added to plates. Plates were observed with invert microscope in 8, 16, 32, 48, and 72 hours after culture.Results: Data implicated that MANNOSE (100 mM) showed the highest effect on increasing cytopathy of Acanthamoeba in HeLa cell culture. Meanwhile, galactose (100 mM) increased the virulence of Acanthamoeba in the cell culture after 32 hours.Conclusion: Adding MANNOSE and galactose to HeLa cell culture contain Acanthamoeba can increase the virulence of the parasite significantly.

Multiple sclerosis (MS) is a complex, demyelinating disease of the central nervous system (CNS) with variable phenotypic presentations, while Guillain-Barre Syndrome (GBS) is the prototypic acute inflammatory disorder that affects the peripheral nervous system. Myasthenia gravis (MG) is a T cell dependent and antibody mediated autoimmune disease. Although it has been shown that complement plays a critical role in the pathogenesis of MS, GBS, and MG, the role of MANNOSE-BINDING lectin (MBL) as a biomarker of immunopathogensis of these diseases and also its association with the severity of them have been poorly investigated. Therefore, in this study we aimed to measure plasma levels of MBL in patients with MS, GBS, and MG.In a case-control study, plasma was obtained from healthy controls (n=100) and also patients with MS (n=120), GBS (n=30), and MG (n=30). Plasma level measurement of MBL was performed using enzyme-linked immunosorbent assay (ELISA).The mean serum level of MBL was significantly different between groups of patients and healthy controls (p<0.001). We also found a positive correlation between plasma levels of MBL and severity scores of MS, MG, and GBS patients including: expanded disability status scale (EDSS) (r=+0.60 and p=<0.001), quantitative myasthenia gravis score (QMGS) (r=+0.56 and p=0.01), and GBS disability scale (GDS) (r=+0.37 and p=0.04).Taken together, our findings suggest that complement activation mediated by MBL contributes to the pathogenesis and also severity of MS, MG, and GBS. However, because the lectin pathway can be involved in several phases of the immune response, further evidence will be required to elucidate the underlying mechanism.