Amifostine inhibits hematopoietic progenitor cell apoptosis by activating NF-kappaB/Rel transcription factors.

We investigated the involvement of NF-kappaB/Rel transcription factors that reportedly can inhibit apoptosis in various cell types in the antiapoptotic mechanism of the cytoprotectant amifostine. In the nontumorigenic murine myeloid progenitor 32D cells incubated with amifostine, we detected a reduction of the IkappaBalpha cytoplasmic levels by Western blotting and a raising of nuclear NF-kappaB/Rel complexes by electrophoretic mobility shift assay. Amifostine inhibited by more than 30% the growth factor deprivation-induced apoptosis, whereas its effect failed when we blocked the NF-kappaB/Rel activity with an NF-kappaB/Rel-binding phosphorothioate decoy oligodeoxynucleotide. In human cord blood CD34(+) cells, the NF-kappaB/Rel p65 subunit was detectable (using immunofluorescence analysis) mainly in the cytoplasm in the absence of amifostine, whereas its presence was appreciable in the nuclei of cells incubated with the cytoprotectant. In 4 CD34(+) samples incubated for 3 days in cytokine-deficient conditions, cell apoptosis was reduced by more than 30% in the presence of amifostine (or amifostine plus a control oligo); the effect of amifostine was abolished in cultures with the decoy oligo. These findings indicate that the inhibition of hematopoietic progenitor cell apoptosis by amifostine requires the induction of NF-kappaB/Rel factors and that the latter can therefore exert an antiapoptotic activity in the hematopoietic progenitor cell compartment. Furthermore, the identification of this specific mechanism underlying the survival-promoting activity of amifostine lends support to the possible use of this agent in apoptosis-related pathologies, such as myelodysplasias.

[The utilization of dry chemistry methods in a campaign to identify risk factor of cardiovascular diseases]. / Uso della chimica a secco in una campagna per L'identificazione dei fattori di rischio per le malattie cardiovascolari.

Dry chemistry methods on a Kodak Ektachem DT 60 instrument have been evaluated during a large scale (60,000 employees of the Italian Telephone Company) screening for the prevalence of cardiovascular risk factors. Comparison between dry chemistry methods on serum samples and liquid phase methods on the same samples yielded the following correlation coefficients: .991 (n = 100) for total cholesterol, .962 (n = 50) for high density lipoprotein (HDL) cholesterol, .987 (n = 100) for triglycerides, .997 (n = 50) for glucose, and .991 (n = 50) for uric acid. A 12% overestimation of serum triglycerides was observed with dry chemistry methods as compared to liquid phase methods. Accuracy and precision of these dry chemistry methods were evaluated by daily (n = 271 days) determinations of all the above parameters on each of two reference sera (low and high concentration). Dry chemistry methods average bias estimates (observed value - expected value)/expected value) x 100, measuring method's accuracy, were: 4.2% for total cholesterol, 1.2% for HDL cholesterol, 3.6% for triglycerides, 3.0% for glucose, 4.6% for uric acid. Average coefficients of variation (standard deviation/mean x 100, measuring method's precision, were: 5.0% for total cholesterol, 6.3% for HDL cholesterol, 5.9% for triglycerides, 3.6% for glucose, 7.2% for uric acid. Dry chemistry methods by Kodak Ektachem DT 60 proved highly comparable with liquid phase methods. Considering their level of accuracy and precision they can be recommended as quick and simple methods for population screenings.

Long term metabolic effects of two dietary methods of treating hyperlipidaemia.

OBJECTIVES: To compare the long term metabolic effects of two diets for treating hyperlipidaemia. DESIGN: Randomised controlled study: after three weeks of normal (control) diet, subjects were randomly allocated to one of two test diets and followed up for six months. SETTING: Lipid clinic of tertiary referral centre in Naples. SUBJECTS: 63 subjects with primary type IIa and IIb hyperlipoproteinaemia entered the study, and 44 completed it. Exclusion criteria were taking drugs known to influence lipid metabolism, evidence of cardiovascular disease, homozygous familial hypercholesterolaemia, and body mass index over 30. INTERVENTIONS: Two test diets with reduced saturated fat (8%) and cholesterol (approximately 200 mg/day): one was also low in total fat and rich in carbohydrate and fibre, and the other was low in carbohydrate and fibre and rich in polyunsaturated and monounsaturated fats. MAIN OUTCOME MEASURES: Fasting plasma lipid and lipoprotein concentrations; blood glucose, insulin, and triglyceride concentrations before and after a test meal. RESULTS: In comparison with the control diet, both test diets induced significant and similar decreases in low density lipoprotein cholesterol concentrations (by a mean of 0.72 (SE 0.15) mmol/l, P < 0.001, for low total fat diet; by 0.49 (0.18) mmol/l, P < 0.05, for high unsaturated fat diet) and plasma triglyceride concentrations (by 0.21 (0.09) mmol/l, P < 0.05, for low total fat diet; by 0.39 (0.15) mmol/l, P < 0.05, for high unsaturated fat diet), while high density lipoprotein cholesterol concentrations after fasting and plasma glucose and insulin concentrations during test meals were not modified by either diet. CONCLUSIONS: Both test diets are suitable (alone or in combination) for treatment of hypercholesterolaemia.