An inhibitor of DNA topoisomerase I from Xenopus laevis ovaries

A novel, heat-resistant and Pronase-sensitive, inhibitor of eukaryotic DNA topoisomerase I has been purified from Xenopus laevis ovaries. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the most purified fraction revealed three bands with apparent molecular masses of 25, 28.5, and 33.5 kDa. The 25- and 33.5-kDa peptides recovered from an SDS-PAGE gel inhibited X. laevis DNA topoisomerase I. The purified inhibitor was specific to DNA topoisomerase I and did not inhibit other DNA enzymes tested. The inhibitor blocked the catalytic activity of DNA topoisomerase I by interacting with the enzyme, rather than by competing for binding sites on substrate DNA. Binding of DNA topoisomerase I to substrate DNA was blocked by the inhibitor, as was the cleavage reaction catalyzed by DNA topoisomerase I. Inhibition of DNA topoisomerase I was relieved by divalent cations Ca2+, Mg2+, or Mn2+.     

Ultrasound‐activated Knoevenagel condensation of malononitrile with carbonylic compounds catalysed by alkaline‐doped saponites

α,β‐Unsaturated nitriles have been synthesized by Knoevenagel condensation of a carbonylic compound with malononitrile, assisted by sonochemical irradiation. Two alkaline‐promoted clays (Li⁺‐ and Cs⁺‐exchanged saponites) have been employed as catalysts. The influence of the carbonylic compound (benzaldehyde or cyclohexanone) and the use of a solvent on the catalytic activity have been studied. Remarkable increase in the conversion values has been found when the reaction is activated by ultrasound, as compared with the thermal activation. In this green, solvent‐free procedure, α,β‐unsaturated nitriles have been produced in very high yields (97%) when the Cs⁺‐saponite is used as catalyst. Copyright © 2004 Society of Chemical Industry     

ALUM SHALE BITUMEN MATURATION AND MIGRATION: IMPLICATIONS FOR GOTLAND'S OIL

Thermal maturation of Swedish Alum Shale kerogen and bitumen has been determined from core samples from Eastern and Central Sweden. In samples from Eastern Sweden (Öland and Gotland), the kerogen and bitumen are thermally immature with respect to petroleum generation. In some areas of Central Sweden (Närke, ÖstergÖtland, and Kinnekulle in VästergÖtland), the kerogen is immature, whereas the bitumen is marginally mature to mature. In other areas of Central Sweden (Halleberg-Hunneberg in VästergÖtland), the kerogen is supermature and the bitumen mature. This suggests that bitumen from a mature source-rock has migrated into the Alum Shales of Central Sweden. Migration in Central Sweden is further evidenced by the occurrence of obviously-migrated bituments in vugs and voids in the organicpoor Ordovician limestone overlying the Alum Shale in Central Sweden, and in concretions within the Alum Shale itself. Based on biomarker distributions of extracted bitumen, Alum Shale kerogen pyrolysate and obviously-migrated oils, and the fact that the Alum Shale in most of the areas studied is the only petroleum source-rock extant, it is suggested that the migrated bitumen in Central Sweden is from the Alum Shale itself. Bitumen has migrated from areas where the Alum Shale is in close proxmity to Permo-Carboniferous intrusions, such as Halleberg-Hunneberg, into nearby areas such as Närke and ÖstergÖtland, where there is no evidence of intrusion and the indigenous organic matter is thermally immature. Other areas, where Alum Shales were associated with intrusions and consequently sourced oil, may have been eroded away. There are producing wells on the island of Gotland, where the Alum Shale is also thermally immature. It is therefore assumed that heating which was responsible for generating Gotland's oil was very localized (such as by an intrusion) or that the oil has migrated from a thermally moremature, distant area. On the basis of reservoir rock porosity, and the fact that the Alum Shale of Gotland contains no migrated component, localized heating is favored.     

Application of a general health policy model in the American health care crisis

There is near consensus that the US health care system requires reform. Only a quarter of the American public has faith in the current system. Health care was one of the major issues considered in the 1992 US presidential election and the search for innovative solutions has transcended administrations.     

Oxidant damage to erythrocyte membrane in glucose-6-phosphate dehydrogenase deficiency: correlation with in vivo reduced glutathione concentration and membrane protein oxidation

Chronic nonspherocytic hemolytic anemia has been observed in a recently described glucose-6-phosphate dehydrogenase (G6PD) variant, G6PDWayne. The mechanical properties of these erythrocytes and other G6PD variants were examined. The deformability of G6PD-deficient erythrocytes was normal, as determined by osmotic scan ektacytometry, and was not significantly affected by hemolytic crisis. In the common varieties of G6PD deficiency, the mechanical stability of the red blood cell (RBC) membrane was greater than normal, but G6PDWayne membranes were abnormally susceptible to shear-induced fragmentation. There was no evidence for a concurrent genetic defect in spectrin, because self-association constants and tryptic digests were normal. The fragility of G6PDWayne membranes appeared to be a consequence of oxidative damage to membrane thiol groups associated with a low glutathione (GSH) level in these RBCs. Associations among GSH level, thiol oxidation, and membrane instability were also found when a larger group of G6PD-deficient RBCs were examined. In normal erythrocytes, 1-chloro-2,4-dinitrobenzene was used to reduce GSH levels by 50%. Membrane thiol oxidation and membrane fragility both increased when these cells were kept at 4 degrees C for 3 to 5 days. Our findings suggest that chronic depletion of GSH leads to the destabilization of membrane skeleton through oxidation of membrane protein thiols.