Apoptosis and cell cycle regulatory effects of adenosine by modulation of GLI-1 and ERK1/2 pathways in CD44+ and CD24- breast cancer stem cells.

OBJECTIVES: Breast cancer stem cells (CSCs) are a small population of tumour cells with the ability of self-renewal and resistance to chemotherapy. Targeting CSCs is a promising strategy for treatment of cancer. A recent study demonstrated that adenosine receptor agonists inhibit glioblastoma CSCs proliferation. At present, the effect of adenosine on breast CSCs has not been reported. Therefore, this study was designed to evaluate the effect of adenosine and its signalling pathways in breast CSCs. MATERIALS AND METHODS: Anti-proliferative effect of adenosine on breast CSCs was evaluated by mammosphere formation and MTS assay. The effect of adenosine on cell cycle progression was examined using flow cytometry. Detection of apoptosis was conducted by Annexin V-FITC. The expression levels of cell cycle and apoptosis regulatory proteins as well as ERK1/2, and GLI-1 were measured by Western blot. RESULTS: Adenosine reduced CSCs population and mammosphere formation in breast CSCs. Adenosine induced G1 cell cycle arrest in breast CSCs in conjunction with a marked down-regulation of cyclin D1 and CDK4. Adenosine also induced apoptosis by regulation of Bax/Bcl-2 ratio, mitochondrial membrane potential depletion and activation of caspase-6. Moreover, adenosine inhibited ERK1/2 phosphorylation and GLI-1 protein expression. CONCLUSIONS: These findings indicated that adenosine induces cell cycle arrest and apoptosis through inhibition of GLI-1 and ERK1/2 pathways in breast CSCs.

Comparison of serum levels of hepcidin and pro-hepcidin in hemodialysis patients and healthy subjects.

Hepcidin prevents absorption of iron from the intestine and inhibits release of iron from macrophages and hepatocytes. For this reason, it seems that high levels of hepcidin are a predisposing factor for anemia in chronic inflammatory conditions such as chronic kidney disease and dialysis patients. This study was designed to determine the role of changes in the level of serum hepcidin in the management of hemodialysis patients. This study included 44 dialysis patients and 44 controls. The hepcidin and pro-hepcidin levels were measured by the enzyme linked immunosorbent assay method. The serum ferritin level was measured by the chemiluminescence method. The mean hepcidin level was 999.3 ± 996.7 ng/mL in the case group and 770.4 ± 815.9 ng/mL in the control group (P = 0.25). The mean pro-hepcidin level was, respectively, 186.1 ± 220.3 pg/mL and 150.87 ± 207.7 pg/mL, in the case group and control groups (P = 0.45). The mean (standard deviation) ferritin level was 816.4 ± 379.4 ng/mL in the case group and 193 ± 171.8 ng/mL in the control group (P < 0.001). In the case group, the correlation between serum ferritin and hepcidin was not significant (r = 0.6, P = 0.08). Also, there was no significant correlation between serum ferritin and pro-hepcidin levels (r = 0.6, P = 0.08). A positive correlation was seen between pro-hepcidin and hepcidin levels (r = 0.92, P < 0.01). In this study, the results showed that the serum hepcidin levels are high in dialysis patients and that there was no correlation with the serum ferritin levels.

Aflatoxin contamination in wheat flour samples from golestan province, northeast of iran.

BACKGROUND: Due to the high toxicity of aflatoxin and its effects on public health, determination of aflatoxin level in Wheat flour samples in the Golestan province, north of Iran was investigated. To examine the effect of seasonal changes, summer and winter sampling was performed with standard sampling methods. METHODS: A total of 200 flour samples were collected from 25 factories. HPLC method with immunoaffinity chromatography was used to measure aflatoxin types (G2, G1, B2 and B1). Statistical analysis was performed by the Pearson correlation test, One-way ANOVA and multivariate regression analysis. RESULTS: Mean total aflatoxin levels of samples were 0.82 and 1.99 ng/g in summer and winter, respectively. Aflatoxin B1 levels were detected in 3.1%, 7.4% over permissible limits by worldwide regulations in samples collected in summer and winter, respectively. Aflatoxins in winter were higher than summer. The highest frequency of aflatoxin contamination in winter was B2 (98%) and in summer G1 (51%). The relationship between humidity and rate of aflatoxin B1 and total aflatoxin was significant in winter. Results of multivariate regression were showed the strongest relationship with humidity and aflatoxin level. Despite the contamination of flour samples, there was no contamination higher than the standard limit of Iran Standard Institute. But it was significantly higher than similar studies from other regions. CONCLUSIONS: Therefore, with regard to negative impacts of aflatoxin on health, aflatoxin contamination should be considered in future programs. Decrease of aflatoxin contamination may be made practical through reducing wheat storage duration and controlling humidity.

Effects of gradual salinity increase on osmoregulation in Caspian roach Rutilus caspicus.

This study was carried out to determine the effects of gradual salinity increase on osmoregulatory ability of the Caspian roach Rutilus caspicus, under conditions which mimic stocking conditions of hatchery-raised fish. Initially, 30 juvenile fish (mean ± S.D. 3.20 ± 0.34 g) were transferred to 20 l circular tanks, in which salinities were changed in a stepwise fashion, from 0 to 5, 10 or 15 at 48 h intervals. The fish at salinity 15 were held for an additional 48 h at this salinity. Forty-eight hours after salinity transfer, survival rate, haematocrit, plasma Cl(-) , Na(+) and K(+) concentrations, osmolality and gill Na(+) /K(+) -ATPase (NKA) activity were measured. The only effect of exposure to 5 was a significant reduction in haematocrit compared to the freshwater control group. Exposure to salinity 10 raised haematocrit, Cl(-) and Na(+) concentrations and osmolality. At 48 h exposure to salinity 15, haematocrit, Cl(-) and Na(+) concentrations and osmolality were significantly higher than freshwater controls, and gill NKA activity was significantly lower, but the effect on NKA was no longer evident at 96 h exposure. There were no effects on survival. These results indicate that R. caspicus juveniles experience an initial non-lethal iono-osmotic perturbation following salinity increase but can adapt to brackish water at salinity 15.

Decreased polyunsaturated and increased saturated fatty acid concentration in spermatozoa from asthenozoospermic males as compared with normozoospermic males.

The lipid composition of the sperm membrane has been shown to exert a significant effect upon the functional quality of spermatozoa. We have studied fatty acid composition of the phospholipids in spermatozoa in asthenozoospermic and normozoospermic men and determined the ratio of polyunsaturated fatty acids (PUFAs) to saturated fatty acids of spermatozoa of these two groups. Fatty acid concentration of spermatozoa was determined in 15 asthenozoospermic and eight normozoospermic semen samples by thin layer chromatography and gas chromatography. The most abundant polyunsaturated and saturated fatty acids in normozoospermic samples were docosahexaenoic acid (DHA 22 : 6 omega3, 98.5 +/- 4.5 nmol per 10(8) spermatozoa, mean +/- SE) and palmitic acid (103 +/- 17 nmol per 10(8) spermatozoa) respectively. The mean +/- SE values of DHA and palmitic acid in asthenozoospermic samples were 53.9 +/- 11.6 and 145 +/- 14.7 nmol per 10(8) spermatozoa respectively. Compared with normozoospermic samples, asthenozoospermic samples showed lower levels of PUFA and higher amount of saturated fatty acids. The mean +/- SE ratios of sperm PUFA/saturated fatty acids in asthenozoospermic and normozoospermic samples were 0.66 +/- 0.06 and 1.45 +/- 0.16 (P < 0.001) respectively. This study demonstrates that spermatozoa of asthenozoospermic men have lower levels of PUFA compared with saturated fatty acids. This may be contributory to the poor motility noted in samples from these men.