Intranasal deferoxamine affects memory loss, oxidation, and the insulin pathway in the streptozotocin rat model of Alzheimer's disease.

Accumulation of metal and the accompanying increase in oxidative stress and inflammation plays an important role in neurodegenerative disease. Deferoxamine (DFO) is a metal chelator found to be beneficial in several animal models of neurodegenerative disease and insult including Alzheimer's disease, Parkinson's disease, stroke, and subarachnoid hemorrhage. In this study, we determine whether intranasally (IN) administered DFO is beneficial in the intracerebroventricular streptozotocin (ICV STZ) rat model of sporadic Alzheimer's disease, which is different from previous models in that it exhibits dysregulation of insulin metabolism as well as oxidative stress and inflammation. Surgical induction of the model included ICV injections of either STZ or citrate buffer (sham in rats), which were treated IN with either saline or DFO (n=10-15/group). Treatment started either before or after injection of STZ to induce the model, and continued throughout the study. IN treatment continued three times per week for three weeks before behavior tests started followed by eventual euthanasia with tissue collection. Spatial memory tests with the Morris water maze showed that STZ rats treated with IN DFO both before and after model induction had significantly shorter escape latencies. Pre-treatment with IN DFO also significantly decreased footslips on the tapered balance beam test. Brain tissue analyses showed DFO treatment decreased oxidation as measured by oxyblot and increased insulin receptor expression. These results further support the potential of IN DFO for use as a treatment for Alzheimer's disease, and show benefit in a non-amyloid/tau rodent model.

Intranasally-administered deferoxamine mitigates toxicity of 6-OHDA in a rat model of Parkinson׳s disease.

Deferoxamine (DFO) has shown therapeutic promise for the treatment of Parkinson׳s disease (PD) as it has reduced both behavioral and biochemical deficits when injected into the brain of rodent models of PD. Intranasally administered DFO targets the brain directly but non-invasively and has been effective in animal models of stroke and Alzheimer׳s disease. In this study we sought to determine whether intranasal (IN) DFO could be neuroprotective for PD in a rat model. PD was induced with a unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle, while sham surgery rats received saline injections. Rats were pre-treated three times with either IN DFO or saline (starting 4 days before 6-OHDA), and post-treated twice/wk for one month before behavioral tests. In the apomorphine-induced rotational test, IN DFO significantly decreased the number of contralateral turns after injection of apomorphine HCl (p<0.05). Also, IN DFO significantly decreased limb asymmetry in the rearing tube as measured with contralateral limb touches (p<0.05). The IN DFO treatment yielded a trend towards decreased contralateral foot-slips on the tapered balance beam, though the difference was not significant. Finally, IN DFO-treated rats had increased preservation of tyrosine hydroxylase immunoreactive neurons in the substantia nigra (p<0.05). These results confirm that DFO is beneficial in a 6-OHDA model and demonstrate improvement in motor deficits and dopaminergic neuronal survival with non-invasive intranasal delivery, making this an attractive potential treatment for PD.

Micromolar levels of intracellular calcium reduce gap junctional permeability in lens cultures.

PURPOSE: To investigate in bovine and embryonic chicken lens cultures the effects of elevated intracellular calcium on the permeability of gap junctions. To determine the changes in intracellular calcium using fura-2. To detect any changes in the phosphorylation of connexin43 after ionophore treatment. METHODS: Lucifer yellow was micro-injected into individual cells, and dye spread to neighboring cells was evaluated. Intracellular calcium levels were measured using the calcium indicator, fura-2. Cultures were also labeled with 32P-orthophosphate followed by immunoprecipitation with antibodies specific for the gap junction protein, connexin43. RESULTS: Bovine lens cultures incubated in the presence of either A23187 or ionomycin showed a reduction in intercellular dye transfer. The intracellular calcium concentrations in bovine cells were increased from a mean value of 0.11 +/- .009 microM in the controls to a mean of 0.40 +/- .073 microM with ionomycin treatment. Subsequent addition of EGTA to the medium decreased the intracellular calcium concentrations to a mean of 0.26 +/- .113 microM and reversed the inhibition of dye spread found with ionomycin. With ionomycin in the medium, the phosphorylated form of connexin43 was not as prominent as in the controls. In contrast, these same treatments had no detectable effect on junctional permeability in the embryonic chicken lens cultures. Dye spread was equally extensive and rapid under control and ionophore conditions, even though fura studies showed an elevation in intracellular calcium levels. CONCLUSIONS: In the bovine cultures, physiologically relevant changes in the levels of cytoplasmic calcium markedly reduced dye transfer. The increase in cytoplasmic calcium was correlated with a change in the phosphorylation level of connexin43. The regulation of junctional communication in the chick lens cultures appears to differ significantly from that in the bovine system.

Effects of insulin and EGF on DNA synthesis in bovine endothelial cultures: flow cytometric analysis.

PURPOSE: To investigate the effects of insulin, epidermal growth factor (EGF), and the corneal storage media--DexSol--at 24 and 48 hours on DNA synthesis in confluent primary cultures of bovine corneal endothelial cells. METHODS: Flow cytometry was used to measure changes in DNA synthesis. This technique allows a large number of cells to be counted and sorted into G1, S, and G2/M phases of the cell cycle. RESULTS: Changing the normal culture media to DexSol had no effect on the cell cycle at 24 or 48 hours. The addition of insulin, EGF, or insulin + EGF to DexSol increased DNA synthesis within 24 hours. The mitotic indices for DexSol, DexSol + insulin, and DexSol + EGF were 0.134 (SE = +/- 0.022), 0.207 (+/- 0.027), and 0.205 (+/- 0.052), respectively. Adding insulin + EGF to the DexSol resulted in the most significant change in S and G2/M, increasing the mitotic index to 0.300 (+/- 0.072) (P = 0.0116). At 48 hours, the presence of the growth factors no longer had any effect. CONCLUSIONS: Flow cytometry was a useful technique in separating cultured bovine corneal endothelial cells according to their DNA content. Analysis of the cultures after the addition of insulin and EGF showed an increase in DNA synthesis. The synergistic effects of the growth factors on corneal endothelial cells suggest that they stimulate mitotic activity by different mechanisms. The addition of mitogens to eye bank storage media may increase corneal endothelial cell densities in donor corneas.

The impact of computed tomography and ultrasonography on the management of patients with carcinoma of the ovary.

We have carried out a prospective study on the impact of computed tomography (CT) and ultrasonography (US) on the management of patients with carcinoma of the ovary. Seventy-eight CT and 88 US scans were performed on 94 patients. Clinicians decided patient management prospectively at the time the CT and/or US was ordered. Clinical assessment differed from the result obtained by CT or US in 45% of cases (35/78 and 40/88, respectively). CT and US altered patient management in only a minority of cases (14/78, 18% and 9/88, 10% respectively). Even when the scan and clinical assessments differed, management was only altered on 14/35 (40%) occasions after CT and on 9/40 (23%) occasions after US, a difference which was not significant. In patients with clinically undetectable disease, management was altered by CT on 17% of occasions and by US on 10%. We conclude that in patients with carcinoma of the ovary CT and US alters patient management in a minority of cases. In view of current financial restrictions in health care, clinicians should be more selective in the use of these imaging techniques. Furthermore, we recommend that similar prospective studies are performed for other clinical situations.