Study of cytotoxic and apoptogenic properties of saffron extract in human cancer cell lines

Saffron (dried stigmas of Crocus sativus L.) has been used as a spice, food colorant and medicinal plant for millennia. In this study cytotoxic effect of saffron extract was evaluated in HepG2 and HeLa cell lines. Meanwhile role of apoptosis and ROS were explored. Malignant and non-malignant cells (L929) were cultured in DMEM medium and incubated with different concentrations of ethanolic saffron extract. Cell viability was quantitated by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). ROS was measured using DCF-DA by flow cytometry analysis. Saffron could decrease cell viability in malignant cells as a concentration and time-dependent manner. The IC50 values against HeLa and HepG2 were determined 800 and 950 μg/ml after 48 h, respectively. Saffron induced a sub-G1 peak in flow cytometry histogram of treated cells compared to control indicating apoptotic cell death is involved in saffron toxicity. This toxicity was also independent of ROS production. It might be concluded that saffron could cause cell death in HeLa and HepG2 cells, in which apoptosis or programmed cell death plays an important role. Saffron could be also considered as a promising chemotherapeutic agent in cancer treatment in future.     

Selective changes in the levels of nicotinic acetylcholine receptor protein and of corresponding mRNA species in the brains of patients with Parkinson's disease

Reductions in the number of neuronal nicotinic acetylcholine receptors (nAChRs) have been shown to occur in connection with Parkinson's disease (PD), but it is still unclear which subtype of this receptor is affected. In the present study we examined various nAChR subtypes employing ligand binding, as well as levels of subunit protein and mRNA in the brains of PD patients and age-matched controls. Binding of [3H]epibatidine and levels of alpha3 mRNA in the caudate nucleus and temporal cortex, but not in the hippocampus were significantly decreased in the PD brain. The level of the alpha3 protein subunit was significantly reduced in all these brain regions but there was no change in the level of alpha4. The level of the beta2 protein subunit in the temporal cortex and hippocampus and the beta2 mRNA in the temporal cortex was lowered. Both the levels of the alpha7 subunit protein and [125I]alpha-bungarotoxin binding were significantly increased in the temporal cortex of PD patients whereas the alpha7 mRNA level was unchanged. These findings reveal selective losses of the alpha3- and beta2-containing nAChRs and an increase in the alpha7 nAChRs that might be related to the pathogenesis of PD.     

Lovastatin stimulates up-regulation of alpha7 nicotinic receptors in cultured neurons without cholesterol dependency, a mechanism involving production of the alpha-form of secreted amyloid precursor protein

The cholesterol-lowering drug lovastatin enhances the secretion of the alpha-secretase cleavage product of amyloid precursor protein (APP). To investigate whether this effect is mediated via activation of alpha7 nicotinic acetylcholine receptors (nAChRs), we treated SH-SY5Y cells and PC12 cells with lovastatin and measured the levels of alpha7 nAChRs, the alpha-form of secreted APP (alphaAPPs), and lovastatin-related lipids, including cholesterol and ubiquinone. The results showed that low concentrations of lovastatin significantly induced up-regulation of alpha7 nAChRs. No effects of lovastatin were observed on alpha3-containing nAChRs, muscarinic receptors, or N-methyl-D-aspartate receptors. alphaAPPs levels increased in the culture medium of cells treated with lovastatin, whereas no change in whole APP was observed. The increase in alphaAPPs was inhibited by prior exposure of these cells to alpha-bungarotoxin, an antagonist of alpha7 nAChRs. The concentrations of lovastatin used in the study did not change the cholesterol content, but high doses can decrease the levels of ubiquinone and cell viability. These results indicate that lovastatin may play a neuronal role that is cholesterol independent. We also show that the up-regulation of alpha7 nAChRs stimulated by lovastatin is involved in a mechanism that enhances production of alphaAPPs during APP processing.     

Rosai-Dorfman disease presenting as multiple soft tissue masses

Rosai-Dorfman disease is a rare clinical disorder which may present in many forms. While classically a disease of lymph nodes, soft tissue involvement is fairly common. Soft tissue involvement can occur without any lymphatic or systemic involvement, and may be difficult to diagnose. We describe a patient presenting with multiple soft tissue masses which on biopsy proved to be isolated cutaneous Rosai-Dorfman disease. MR imaging showed two well-defined nonspecific superficial masses that enhanced intensely. Review of the literature suggests that when this disease presents in soft tissue, multiple foci of involvement may be common. Although rare, Rosai-Dorfman disease should be considered in the differential diagnosis of patients presenting with multiple soft tissue masses.     

Thyroglobulin increment after thyroid hormone withdrawal is a reliable indicator for the detection of significant remnants or metastases in patients with differentiated thyroid carcinoma

Serum thyroglobulin (Tg) measurement has a pivotal role in the management of differentiated thyroid carcinoma (DTC). Serum Tg increment after thyroid hormone discontinuation seems to be a better predictor of tumor recurrence, however, minimal Tg increment may not be a specific marker. This study tries to evaluate the importance of different levels of Tg increment after thyroid hormone discontinuation. Fifty-five patients (46 females and 9 males with mean age of 41.40 yrs) with DTC, treated with total or subtotal thyroidectomy and radioiodine-131 ((131)I) were studied. Ninety-one per cent of the patients had papillary carcinoma. Serum Tg and thyroid stimulating hormone (TSH) were measured using high sensitive IRMA assays during thyroxine (T4) suppression and after discontinuation of T4 treatment. The mean time interval between Tg on T4 and off T4 was 110.29+/-53.43 days and less than 180 days in all patients. Serum Tg level was increased >or= 1 ng/ml in 25 patients after discontinuation of T4. Of these patients, 17 had metastatic disease or a detectable thyroid remnant. Of 16 patients with unchanged Tg (-1or= 7 ng/ml had residual disease or metastases. If DeltaTg was unchanged or decreased, the negative predictive value was 83.3%. The sensitivity of WB(131)IS was 63.6% for the detection of thyroid remnant or metastases. Our study indicates that DeltaTg is a more reliable indicator of remnant disease than on T4-Tg or off T4-Tg levels.     

Metastatic burst fracture risk prediction using biomechanically based equations

Clinical guidelines are a useful adjunct to select patients with spinal metastases for prophylactic intervention. The objective of this study is to determine the ability of biomechanically based models to accurately predict metastatic burst fracture risk. Ninety-two vertebrae with osteolytic spinal metastases were examined retrospectively. Vertebrae were categorized as burst fractured, wedge fractured, or intact and analyzed using three predictive models: vertebral bulge (maximum radial displacement under load), vertebral axial displacement (maximum axial displacement under load), and a volumetric estimate of tumor size. The load-bearing capacity parameter (tumor volume, bone mineral density, disc quality, pedicle involvement) was determined from computed tomography while the load-bearing requirement parameter (pressure load, loading rate) was determined using computed tomography and patient records (retrieved for 37 patients [52%]). Fracture prediction was optimized using the vertebral bulge model considering only load-bearing capacity with a specificity, sensitivity, and confidence interval of 1 to yield a clear threshold for burst fracture risk. Fracture prediction in the other two models, vertebral axial displacement considering only load-bearing capacity and tumor size, also was strong with receiver-operator curve values of 0.992 and 0.988, respectively. The predictive power of these models can provide useful clinical information for prophylactic decision-making.     

Reduced levels of Abeta 40 and Abeta 42 in brains of smoking controls and Alzheimer's patients

The effects of nicotine on levels of Abeta 40 and Abeta 42 and nicotinic receptor binding sites were studied in brains from nonsmoking and smoking patients with Alzheimer's disease (AD) and aged-matched controls. The levels of soluble and insoluble Abeta 40 and Abeta 42 in frontal cortex and Abeta 40 in temporal cortex and hippocampus were significantly decreased in smoking AD patients compared to nonsmokers with AD. In smoking controls the levels of soluble and insoluble Abeta 40 and Abeta 42 in the frontal and temporal cortex were significantly lower than in nonsmoking controls. The binding of [(3)H]cytisine in temporal cortex was significantly increased in smokers with AD compared to nonsmokers with AD. In smoking controls [(3)H]cytisine and [(3)H]epibatidine binding were significantly increased from 1.5- to 2-fold compared to nonsmoking controls whereas binding sites for [(125)I]alpha-bungarotoxin was less up-regulated. These results indicate that selective nicotinic receptor agonists may be a novel protective therapy in AD by reducing Abeta levels as well as the loss of nicotinic receptors in AD brain.     

Direct determination of triamterene by potentiometry using a coated wire selective electrode

A coated wire triamterene-selective electrode based on the incorporation of a triamterene–tetraphenylborate ion-pair in a poly(vinylchloride) coating membrane was constructed. The influence of membrane composition, temperature, pH of the test solution, and foreign ions on the electrode performance were investigated. The electrode showed a Nernstian response over a triamterene concentration range from 1.0×10⁻⁶ to 3.5×10⁻² M, at 25 °C, and was found to be very selective, precise, and usable within the pH range 4.5–7.5. The standard electrode potentials, E°, were determined at 15, 20, 25, 30, 35, 40 and 45 °C and used to calculate the isothermal temperature coefficient (dE°/dt) of the electrode. Temperatures higher than 45 °C seriously affected the electrode performance. The electrode was successfully applied to the potentiometric determination of triamterene hydrochloride both in pure solutions and in pharmaceutical preparations.