Comparison of nucleostemin gene expression in CD133+ and CD133- cell population in colon cancer cell line HT29.

BACKGROUND: Nucleostemin has been shown to be essential for proliferation and survival of colon cancer cells. In this study, we evaluate and comparing nucleostemin expression in CD133+ and CD133- colon cancer cell line HT29. MATERIALS AND METHODS: After preparation and culturing of HT29 cell line, isolation was performed using magnetic cell separation system by CD133 MicroBeads and phycoerythrin conjugated to monoclonal anti-human CD133 monoclonal antibody and analyzed with flow cytometry. For quantitative expression of nucleostemin in HT29, CD133+ and CD133- cells used specific nucleostemin primer and glyceraldehyde 3-phosphate dehydrogenase primer as endogenous control. RESULTS: The results showed the percentage of CD133+ cells in HT29 colon cancer cell line ranged from 36.5% to 41.5%, whereas the percentage of CD133- cell ranged from 58.5% to 63.5%. The expression rate of nucleostemin in HT29, CD133+ and CD133- cells were 1.44 ± 0.78, 1.60 ± 0.70 and 1.00 ± 0.18 (respectively). The comparison of expression rate represents no significant difference in nucleostemin expression in CD133+, CD133- and HT29 colon cancer cells. CONCLUSION: It is concluded that nucleostemin expression could not be specific in a certain type of cells in colon cancer cell line HT29 and controlling strategies in colon cancer must not be focused on one certain type of colon cancer cells as main expressing nucleostemin gene.

Identification of shiga toxin and intimin genes in Escherichia coli detected from canary (Serinus canaria domestica).

The pathogenicity of enterohemorrhagic Escherichia coli (EHEC) strains is, in large part, due to shiga toxin (Stx) genes (Stx1 and Stx2) and/or intimin (eae) gene. The purpose of this study was to analyze the role of domestic canaries (Serinus canaria domestica) as a reservoir of Stx and intimin producing strains of E. coli. For this study, a total of 50 cloacal swabs were collected from 50 healthy domestic canaries. Cloacal swabs were cultured and tested using standard methods of microbiology. After primary identification of E. coli, DNA was extracted and polymerase chain reaction was performed using specific primers for Stx1, Stx2 and eae genes. In this study, three of 50 samples were found to be Stx2 positive. In the present study, nine (18%) of 50 canaries tested were positive for eae gene. Only 2% of total canaries tested were positive for simultaneous Stx and eae genes. By considering the presence of Stx genes in E. coli isolated from cloacal contents of canary, this hypothesis expressed that the canaries may be the carriers of virulence genes that can risk human health. Canary was considered to be a reservoir of Stx and intimin genes and make these birds important vehicles for the spread of zoonosis infection.

Effect of nanosilver on blood parameters in chickens having aflatoxicosis.

This experiment is designed to investigate the positive effects of commercial nanosilver compound on blood parameters in experimental aflatoxicosis in broiler chickens. For this, 270 one-day-old broiler chickens were randomly divided into six treatment groups with three replicates. The experimental groups were group A: chickens fed with basal diet; group B: chickens fed with 3 ppm productive aflatoxin in basal diet; groups of C, D, E and F received Mycoad (2.5 g/kg diet), Mycoad (2.5 g/kg diet) + productive aflatoxin (3 ppm), Nanocid (2500 ppm), and Nanocid (2500 ppm) + productive aflatoxin (3 ppm) in basal diet, respectively. Results revealed that some of the blood parameters such as mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration lymphocytes, neutrophils, basophils, monocytes, and eosinophils percentage were not affected in this experiment; whereas, hemoglobin percentage and white blood cell (WBC) count in all the groups fed with 3 ppm aflatoxin except nanocid + aflatoxin decreased significantly (p < 0.05). There are no significant differences between the groups that received nanocid + aflatoxin and mycoad + aflatoxin in hemoglobin percentage and WBC count parameters. The red blood cell count and hematocrit in chickens received aflatoxin were significantly lower than other groups (p < 0.05). Therefore, this study suggests that nanocid similar as mycoad can be useful in reducing the adverse effects of aflatoxin on blood parameters in chickens affected with aflatoxicosis.

Molecular detection of avian pox virus from nodular skin and mucosal fibrinonecrotic lesions of Iranian backyard poultry.

In recent years, some outbreaks of skin lesions suspected to be avian pox were observed in the backyard poultry in different parts of western areas in Iran. Consequently, 328 backyard poultries with suspected signs of avian pox virus infection were sampled. All birds showed nodular lesions on unfeathered head skin and/or fibronecrotic lesions on mucus membrane of the oral cavity and upper respiratory tract. For histopathological analysis, the sections of tissue samples from cutaneous lesions of examined birds were stained with H&E method. For PCR, after DNA extraction a 578-bp fragment of avian pox virus from 4b core protein gene was amplified. Results showed 217 and 265 out of 328 (66.1 and 80.7%, respectively) samples were positive for avian pox virus on histopathological and PCR examination, respectively. In this study, the samples that had intracytoplasmic inclusion bodies on pathologic examination were PCR positive. This study revealed that PCR is a valuable tool for identification of an avian pox virus and that the frequency of pox infection in backyard poultry in western areas of Iran is high.

Nanosilver effects on growth parameters in experimental aflatoxicosis in broiler chickens.

Aflatoxicosis is a cause of economic losses in broiler production. In this study, the effect of one commercial nanocompound, Nanocid (Nano Nasb Pars Co., Iran) was evaluated in reduction of aflatoxin effects on the growth and performance indices in broiler chickens suffering from experimental aflatoxicosis. For this, a total of 300 one-day-old broiler chicks (Ross strain) were randomly divided into 4 groups with 3 replicates of 15 chicks in each separated pen during the 28-day experiment. Treatment groups including group A: chickens fed basal diet, group B: chickens fed 3 ppm productive aflatoxin in basal diet, group C: chickens fed basal diet plus 2500 ppm Nanocid, and group D: chickens fed 3 ppm productive aflatoxin and 2500 ppm Nanocid, in basal diet. Data on body weight, body weight gain (BWG), feed intake, and feed conversion ratio (FCR) were recorded at weekly intervals. Also cumulative data were assessed. Results showed, although supplement of Nanocid to conventional diet had no effect on performance but addition of Nanocid to diet containing 3 ppm aflatoxin increased significantly the cumulative BWG, cumulative feed consumption and decreased FCR in the last 2 weeks of experimental period. The improvement in these performance indices by supplement of Nanocid to diet containing aflatoxin showed the ability of Nanocid to diminish the inhibitory effects of aflatoxin.