Differential pulse polarography for monitoring cell cultures

The differential pulse polarography has been used on samples taken from the medium during the culture of chinese hamster ovary cells with a view to producing the active principle of an anti-cancer drug. An electrochemical signal at a potential of −1.8 V with respect to the saturated calomel electrode was correlated with the lactic acid concentration, the pH variation and the number of cells. The peaks observed in oxidation at potentials −0.35 V and −0.05 V were associated with the transformation of the cystine present in the nutrient medium into cysteine and provide an indication of the modification of the reducing power of the medium during culture. No correlation was found between the electrochemical behaviour and cell production.     

Induction of metallothionein-like proteins in the digestive gland of Mytilus galloprovincialis after a chronic exposure to the mixture of trace heavy metals

Abstract

The amount of metallothionein-like proteins (MTLPs) induced in the digestive gland of exposed and control specimens of Mytilus galloprovincialis is measured after the 14 days of the experiment. Three groups of mussels were kept in a flow-through seawater. The amount of induced MTLPs has been measured in the digestive gland of mussels exposed to the added Cd and, separately, to the added mixture of Cd, Cu, Pb while Zn was for all three experimental groups enhanced to 11.80 μg L^?1 in flow-through seawater, as the consequence of the contamination from the pipeline for bringing the seawater to the aquarium. The induced amount of MTLPs in exposed mussels is compared with its amount in control specimens, which were kept for 14 days in the flow-through seawater at the natural concentration level of Cd, Cu, Pb and enhanced Zn concentration, as previously explained. The metal concentrations in seawater to which mussels were exposed for 14 days were of Cu 3 times, of Pb 7 times, of Zn 20 times higher than their natural concentration level. Only the concentration of Cd was 100 times higher than the natural one. Therefore, the presented results are applicable to understanding the effect of the mixture of environmentally relevant metals on the induced amount of MTLPs as the indicator substance of seawater pollution.     

Prolonged Lag in Population Outbreak of an Invasive Mussel: A Shifting-Habitat Model

Biological invasions pose a great threat to the integrity of natural communities. Some invasive species demonstrate a population explosion shortly after arrival while in other cases a prolonged lag between arrival and population outbreak is evident. This paper describes a case of a prolonged lag and explores the possible mechanism for this lag. The Red Sea mussel Brachidontes pharaonis, a Lessepsian migrant, was first recorded in the Mediterranean seven years after the opening of the Suez Canal in 1869. Since then it spread along the Israeli coast and as far northwest as Sicily. Studies conducted in the late 1970s, when B. pharaonis was still rare, predicted that it would not establish dense populations along the Israeli coast and would not outcompete the indigenous mussel Mytilaster minimus, although it has strong negative effects on survival and growth of the native species. It was attributed to the invader's low intrinsic rate of increase relative to that of the native species, and to strong density-independent mortality generated by exposure to high wave action and sedimentation. In contrast to these predictions, we found massive formations of B. pharaonis beds after lag of about 120 years. We looked for distributional patterns that may explain this lag and found no south–north gradient but a strong habitat-dependent colonization. Most apparent are dense B. pharaonis mussel beds (density >300 per 100cm²) on rocky platforms where mussel beds were absent in the past. These platforms lack the vermetid rim that is typical to this formation. In platforms protected by a biogenic rim, sediment accumulation is high and perennial algae flourish. None of the mussel species form beds in such habitats. We suggest that the delayed formation of B. pharaonis beds along the Israeli coast is a consequence of a recent shift in habitat conditions on some platforms. It is possible that receding of the biogenic rim at the edge of these platforms allowed more effective washing, reduced sediment accumulation, and reduced perennial algae cover making platforms more suitable for the mussels. Lower density-independent mortality allowed B. pharaonis to dominate on such platforms over the indigenous species. On beachrock, a habitat previously dominated by M. minimus, we recorded a rapid shift in numerical domination to B. pharaonis (from 1:7 to 1.4:1 Brachidontes/Mytilaster individuals) over a period of 4 years (1995–1999). This is probably a result of saturation of the habitat by B. pharaonis recruits originating from the established populations on platforms. Salinity changes and a potential genetic shift may also have contributed to the invasive mussel outbreak.     

Simultaneous determination of Zn,Cd, Pb,and Cu in urine of patients with blackfoot disease using anodic stripping voltammetry

Blackfoot disease (BFD) is an endemic peripheral vascular disorder resulting in gangrene of the lower extremities, especially the feet, among residents in a limited area on the southwest coast of Taiwan. In the present study, the concentrations of zinc, cadmium, lead, and copper in urine of BFD patients with matched normal controls are investigated by differential pulse anodic stripping voltammetry (DPASV) on a hanging mercury drop electrode (HMDE). The analytical results indicate that urinary copper, cadmium, and lead of the BFD patients are significantly higher than those of the controls. In addition, the patients showed a significantly lower concentration of zinc in the urine than the normal controls. The possible connection of these elements with the etiology of the disease is discussed.     

Chemiluminescence Emission during Reactions between Superoxide and Selected Aliphatic and Aromatic Halocarbons in Aprotic Media

The reactions between superoxide free radical anion (.O⁻₂) with the halocarbons CCl₄, CHCl₃, BrCH₂CH₂Br(EDB), decachloro-biphenyl (DCBP), and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in dimethyl sulphoxide (DMSO) results in the emission of chemiluminescence (CL). The chemiluminescence reactions are characterized as having biphasic second order kinetics, CL wavelengths between 350 nm and 650 nm, and exhibiting perturbation by chemicals reactive with singlet oxygen. These data suggest that singlet oxygen species are the excited state responsible for the light emissions. Polarographic studies confirm .O⁻₂ consumption and halide release in the reactions, while gas liquid chromatography and NBT reduction demonstrate the decomposition of the halocarbons into products. A chemiluminescent reaction mechanism is proposed involving reductive dehalogenation of the halocarbons and the generation of singlet oxygen. The significance of singlet oxygen generation is discussed with respect to a general mechanism for explaining the rapid initiation of lipid peroxidative membrane damage in halocarbon toxigenicity in animal and plant tissues.     

Studies on the effects of the antitumor agent camptothecin and derivatives on DNA. Camptothecin potentiated cleavage of DNA by bleomycin in vitro

Addition of sodium camptothecin (2a, Fig. 1) in comparable low concentrations to the glycopeptide antitumor antibiotic bleomycin (BLM) leads to enhanced rates of single-strand scission of PM2-covalently closed circular DNA, whereas sodium camptothecin alone has no effect. A similar enhancement of DNA scission by sodium camptothecin is produced with the 1 : 1 bleomycin-iron complex alone or in conjunction with NADPH as an additional reductant. The interpretation that camptothecin may substitute for the reducing requirement of the antibiotic is supported by its oxidation at 37°C by the 1 : 1 bleomycin iron complex, by iron salts or more efficiently by hydrogen peroxide to the known hemiacetal (3, Fig. 1).Electrochemical studies of 2a, its analogues and selected model compounds established that the α-pyridone ring D is most susceptible to a one-electron reduction at a reversible potential of ?0.95 ± 0.01 V. The reduced camptothecin is a transient species readily capable of donating an electron. This process may by compatible with a coupled reduction of the sequestered Fe(III) in the glycopeptide antibiotic necessary for the expression of antibiotic and antitumor properties. The results may provide a mechanistic rationale for the observed potentiation of the antitumor activity of bleomycin by camptothecin in vivo.