hassan morovati

One of the most important complications of toxoplasmosis is its early diagnosis. It seems that GRA7 protein can be a good candidate for detection of the acute phase in Toxoplasmosis. Accordingly, the present study aimed to diagnose toxoplasmosis via a newly immunochromatographic test using recombinant antigen gra7.

The parasite was cultured in mice and then were used for DNA extraction. The gra7 gene was amplified by PCR and cloned into the pET-32a (+) plasmid. Thereafter, the recombinant vector was transferred into the Escherichia coli Rosetta strain and gra7 was detected via SDS-PAGE and western blotting. The bacterial lysate was used to purify the protein by Ni-NTA affinity chromatography .Anti-human gold conjugated antibody, test line and control line were injected to conjugate pad and nitrocellulose membrane, respectively, and all the layer were assembled. By using serum of patients and healthy individuals, manufactured kits were evaluated.

Our results indicated that the selected gene was correctly cloned and the protein of interest was produced and purified. The test revealed sensitivity and specificity of 100 and 96.7 percent, respectively. The kit was also shown to be stable over 16 weeks in 37°C.

The choice of antigen based on cellular and clinical features of the parasite, as well as the use of previous outcomes yielded to develop a rapid diagnostic test for toxoplasmosis.

Our previous studies revealed an inhibitory effect of ICD-85; a venom derived peptide has been extracted by researchers of Razi vaccine and serum research institute, on breast cancer cell line (MDA-MB231). The present study was undertaken to evaluate the anti proliferative effect of HL60 cells. The effect of ICD-85 on proliferation of HL-60 cancer cells was determined using the MTT assay. A quantitative cytotoxicity assay based on the release of lactate dehydrogenase (LDH; EC 1.1.1.27) was performed 24 h after exposure to various concentrations of ICD-85. The morphological changes of ICD-85 treated HL-60 cancer cells were observed under electron microscope. ICD-85 induced marked concentration inhibition of cancer cell proliferation with an IC50 value of 0.04µg/ml following 24 h incubation. Electron microscopic findings of ICD-85 treated cells showed nuclear material condensation, endoplasmic reticulum dilation, mitochondria swelling or degradation, increased cytoplasmic vacuoles, reduction or disappearance in cytoplasmic process and decreased nuclear/cytoplasmic ratio. LDH determination of cultured media of HL60 cells exposed to various concentration of ICD-85 revealed no significant changes in LDH levels compared with untreated cells. The results of present study indicated that ICD-85 inhibited the cancer cell proliferation by inducing apoptosis rather than necrosis.

Pneumocystis pneumonia (PCP) has been historically the most prevalent opportunistic infection in patients infected with the human immunodeficiency virus. Culture of the organism has not been faced with suitable success in artificial media, while various results have been reported for cell culture media. The aim of this study was proliferation of Pneumocystis carinii on the Razi Bovine Kidney (RBK) cell line and to compare growth rate with ‘Vero’ and ‘MRC-5’ cell lines. We used 6 rats (Sprague-Dawley) provided from Razi Institute to infect with Pneumocystis carinii after suppressing the immune system with methylprednisolone acetate (40 mg/kg). Methylprednisolone acetate was used subcutaneously once a week for 8 weeks. Samples were homogenized after separation of the lung tissue. Microscopic examination was applied for prepared smears to confirm the presence of Pneumocystis carinii. Purified trophozoites were then inoculated into the cell line flasks. Growth rate was estimated by counting the trophozoite in each day. Number of cultivated organisms was increased after 5 days incubation in all applied cell lines. Growth rate of Vero, MRC-5 and RBK were 3, 3, and 3.75 times more respectively in comparison with number of the calculated cells in first day. Hence the difference between RBK and two other cell lines was significant (p = 0.023). RBK cell line is suitable to proliferate Pneumocystis carinii.